Dissertations / Theses on the topic 'Lymphatic System, drug effects'

A current initiative to eliminate lymphatic filariasis (LF), headed by the World Health Organization, aims to interrupt transmission of the disease through yearly community-wide treatment with the broad spectrum anthelmintic albendazole (ABZ), in combination with ivermectin (IVM) or diethylcarbamazine (DEC). Over the years, the use of both ABZ and IVM in the treatment of veterinary parasites has led to widespread anthelmintic resistance against these drugs. In this study, we genotyped microfilaria of Wuchereria bancrofti, a causative agent of LF, in order to detect the presence of mutations which confer ABZ resistance in other parasites, and we identified such mutations in worms obtained from untreated patients in Ghana and Burkina Faso, West Africa. Microfilaria from patients who had been treated with ABZ + IVM, had a significantly higher frequency of the resistant genotype, and this frequency was even higher in worms from patients that had received two rounds of treatment. In addition, the untreated population of microfilaria had an excess of homozygotes in the population. This excess homozygosity was equivalent to a Wright's Inbreeding Statistic of FIT= 0.44, and we found that the population was significantly subdivided between patients. In order to better understand the mechanisms and factors involved in the potential spread of ABZ resistance, caused by such mutations, through a population of Culex-transmitted W. bancrofti, we developed a deterministic model that incorporates genotype structure into the epidemiological model EPIFIL. This model predicts that the combination of ABZ + DEC leads to stronger selection for the resistant genotype than ABZ + IVM, and that drug efficacy assumptions are an important factor affecting the spread of drug resistance. Treatment coverage, non-random mating, initial allele frequency and number of treatments also had substantial impact on the speed and magnitude of the spread of ABZ resistance. When we expanded this model to include potential IVM-resistance alleles we found that, under ABZ + IVM treatment, selection for resistance to either drug is enhanced by the presence of resistance against the second drug. Similarly, excess homozygosity caused by parasite non-random mating may increase selection for a dominant IVM resistance allele through enhancing the spread of a recessive ABZ resistance allele. Resistence developed more slowly when it was inherited as a polygenic trait. Results from this study suggest that resistance monitoring is crucial, as resistance may not be apparent until treatment is stopped, recrudescence occurs and treatment is reapplied.

Since type B adverse drug reactions tend to be rare and serious, they tend to be treated by tertiary-care specialists; and since they are commonly iatrogenic, the specialists should be concerned to document carefully not only the case per se but also the drug use history, leading to practice data of good research quality. The specialist should also be concerned to submit the data record to a central facility that would supply the probabilities, evidence-based, that a recent drug use by a patient caused the adverse event. Continual and systematic accumulation of these data records at the central facility--using the same logistic and organizational framework for each of different type B events--provides for both the numerator and denominator series for etiologic research. Since the targeted events are quite rare, the catchment population of the "registry" would have to be very large, international in scope, especially if the system is to provide for rapid resolution of crises arising from novel suspicions of type B effects with respect to newly marketed drugs.

Vitamin D regulation is associated with several human disorders and contributes to various cellular mechanisms. Calcitriol (commercially available as Rocaltrol), an active Vitamin D metabolite, is known as a neuro-protective and anti-cancer drug but most importantly helps maintaining the calcium homeostasis inside the human body. The effectiveness of calcitriol to perform as an effective therapeutic agent is counteracted by its calcemic effects. In order to obtain better therapeutic results, synthetic calcitriol analogs without these calcemic effects have been recently developed but they are not yet cost-effective and their production is time-consuming. In order to determine the best active form of calcitriol that could provide higher chemotherapeutic activity without these calcemic effects, calcitriol mode of action was studied using yeast as a model system. In order to achieve this, we analyzed the calcitriol effects on yeast cellular growth based on calcium intake levels. In this work, we also assessed yeast strains with gene deletions of selected calcium transporter genes to understand the calcitriol metabolism. For the aim of understanding hypercalcemic effects of calcitriol, we developed a hypothesis based on calcitriol interactions with oxygen. Interestingly, use of an anaerobic model validated the oxygen interactions with calcitriol that might possibly cause calcemic effects on patients. Anaerobically grown yeast treated with calcitriol showed significantly less intracellular calcium levels when imaged under indo-1 calcium binding fluorescence dye as compared to calcitriol treated yeast grown under aerobic conditions. Finally, we predict that calcitriol might control free radical generation within the yeast system based on experiments with AAPH and UV- irradiation.

Dissertação para obtenção do Grau de Mestre em Biotecnologia
The increased incidence of type II diabetes has emerged as a major concern in controlled human immunodeficiency virus (HIV)infection. There is a general lack of data to support the best combined antiretroviral therapy (cART) option to treat HIV-patients with pre-diabetes and nevirapine has been described has a glucose-friendly antiretroviral. On the other hand, it is known that diabetes could influence the pharmacokinetics of several drugs. This aspect is particularly relevant for drugs with narrow therapeutic window, which is the case of nevirapine. To understand if nevirapine is a good choice for pre-diabetic HIV-patients, the effect of insulin resistance in NVP pharmacokinetics as well as the effect of nevirapine on insulin resistance, fasting glycemia and mean arterial pressure was evaluated. Moreover, nevirapine effect on thiols content, an endogenous antioxidant defence system, was also evaluated. To achieve the main goal four groups of female Wistar rat were used: a control group, a control group treated with nevirapine, an insulin resistant group and an insulin resistant group treated with nevirapine. An influence of a pre-diabetic status on nevirapine pharmacokinetic was found. Nevirapine and its phase I metabolites presented changes in disposition and the metabolite profile pattern was changed. Moreover, nevirapine, in a pre-diabetic perspective, is associated with a beneficial effect on fasting glycemia, while it has no effect on sensitivity to insulin or in arterial pressure. Furthermore, nevirapine is associated with a lower degradation of total glutathione. Nevirapine might be a good option for HIV-infected patients at higher risk of develop diabetes or in pre-diabetic condition. Moreover, while further studies are necessary to consolidate this issue, nevirapine might be less toxic in pre-diabetes. Although, the decreased bioavailability of nevirapine in pre-diabetes requires special attention, as an adjustment of nevirapine dose might be required in this subpopulation.
Portuguese Foundation for Science and Technology - PTDC/QUI-QUI/113910/2009, PTDC/SAU-ORG/111417/2009

Abstract The rationale for automatically monitoring anesthetic drug effects on the central nervous system (CNS) is to improve possibilities to gain objective information on a patient's state and to adjust the medication individually. Although monitors have shown their usefulness in practice, there are still a number of unclear issues, especially with respect to the scientific foundations and validity of CNS monitoring techniques, and in monitoring the light hypnotic levels. Current monitors are, for example, often based on heuristics and ad hoc solutions. However, a quantitative index for anesthetic drug effect should have a sound relationship with observations and with the selected control variable. The research objectives are: (1) to explore propofol anesthetic related neurophysiological phenomena that can be applied in the automatic assessment of CNS suppression; (2) to develop a valid control variable for this purpose; (3) by means of digital signal processing and mathematical modeling, to design and to evaluate the performance of an index that correlates with the control variable. This dissertation introduces potentially useful neurophysiological phenomena, such as changes in phase synchronization between different EEG channels due to anesthesia, and painful stimulus evoked responses during the burst suppression. Furthermore, it refines the progression of the time-frequency patterns during the induction of anesthesia and shows their relation to the instant of unresponsiveness. The presented spontaneous and evoked EEG phenomena provide complementary information about the CNS functional suppression. Most significantly, the dissertation proposes a continuous and observation based control variable (r scale) and the means to predict its values by using EEG data. The definition of the scale provides a basis for anticipating the instant of the loss of consciousness. Additionally, the phase synchronization index as an indicator of drug effect is introduced. The approximate entropy descriptor performance is evaluated and optimised with a non-stationary signal recorded during the induction of anesthesia. The results open up opportunities to improve the preciseness, scientific validity and the interpretation of information on the anesthetic effects on CNS, and therefore, to increase the reliability of the anesthesia monitoring. Further work is needed to extend and verify the results in deep anesthesia.

Myelin and oligodendrocytes (OLGs) are the apparent targets of the immune-mediated injury that underlies the development of multiple sclerosis (M8). Recovery from M8 clinical relapses likely reflects remyelination attributed to recruitment and differentiation of oligodendrocyte progenitor cells (OPCs), rather than to new process formation by previously myelinating OLGs. Newly emerging M8-directed immuno-modulatory therapies (statins and FTY720) can readily cross the blood-brain barrier and have been shown to impact signaling pathways implicated in cytoskeletal regulation, differentiation, migration, and survival; these are cellular events presumably important for myelin integrity and remyelination.
Statins inhibit the production of cholesterol (concentrated in the myelin membrane) and isoprenoids (post-translational attachments regulating the functions of proteins such as the Rho GTPases). We showed that treatment of human and rodent-derived OPCs with lipophilic statins induced an initial process extension associated with enhanced differentiation and impaired spontaneous migration, whereas prolonged treatment induced process retraction and cell death. Rodent and human mature OLGs demonstrated similar cytoskeletal and survival responses. Chronic simvastatin therapy of mice inhibited remyelination following demyelination induced by the OLG toxin, cuprizone, attributed to a block in OPC differentiation and consequent decrease in mature OLGs. Even fully myelinated animals treated with simvastatin over the long-term demonstrated a decrease in myelin in the brain by maintaining oligodendroglial cells in the pre-OLG state and preventing continual replacement of mature OLGs.
FTY720 is an agonist of G-protein-coupled receptors S1P1, 3, 4, and 5, that are associated with distinct receptor isotype-selective activation of Rho GTPases. In human OPCs, FTY720 could induce initial S1P3/5-dependent process retraction associated with an inhibition of differentiation, and subsequent S1P1-dependent process extension. Mature OLGs showed a dose-dependent cyclic modulation of process extension and retraction was observed over time. Both human OPCs and OLGs were rescued by FTY720 under death-promoting environments. Both cell types also demonstrated a cyclic and reciprocal modulation of S1P1 and S1P5 mRNA levels, reflected in the recurring receptor isotype-dependent functional responses over time. Studies using organotypic cerebellar slice cultures demonstrated that FTY720 did not impact myelin integrity under basal conditions, yet accelerated remyelination following lysolecithin-induced demyelination. Both treatment regimens were associated with an extension of OPC and mature OLG processes.
Our observations demonstrate that drug concentrations used to modulate immune function can have differential dose and time-dependent effects on OPCs, OLGs, as well as on myelin and remyelination processes. Our findings indicate the need to monitor the effects of putative immuno-modulatory therapies on myelin-related processes in MS patients.

Methylenedioxymethamphetamine (MDMA) popularly known as "Ecstasy" continues to gain popularity as a recreational drug that has been shown to increase serotonin and dopamine levels. The present study has demonstrated that repeated exposure to MDMA produces long-term damage to serotonergic and dopaminergic neurons in various regions of the rat brain.

Objetivo Avaliar os efeitos do tabagismo sobre o sistema cardiovascular, através de metodologia não invasiva, analisando as alterações hemodinâmicas clínicas e propriedades elásticas arteriais, após exposição ao fumo. Métodos Estudo realizado em 45 voluntários sendo 18 (F) e 27 (M), fumantes T (n = 25, idade 40 ± 9 anos) e não fumantes NT (n = 20, idade 39 ± 9 anos), que foram submetidos a determinação do monóxido de carbono (CO) e avaliação das propriedades elásticas arteriais por ultrassonografia e teste de função endotelial (hiperemia reativa - HR) antes (B1) e após (B2) a exposição por 20 minutos a um cigarro ou à degustação de uma bala. Resultados Os grupos NT e T mostraram-se homogêneos. A concentração de CO no grupo T era maior do que no grupo NT em condições basais. A concentração de CO no grupo NT não variou antes e após à exposição proposta (bala) mas no grupo T aumentou de maneira significante após o cigarro. A pressão arterial sistólica (PAS) foi igual no B1 nos dois grupos, mas significativamente maior no B2 para o Gr T. A pressão arterial diastólica (PAD) no B1 e B2 não mostrou variação estatísticamente significante nos dois grupos. A freqüência cardíaca (FC) apresentou comportamento estatisticamente diferente nos dois grupos (NT apresentou redução e T apresentou aumento). Em relação às propriedades elásticas arteriais e resposta do fluxo regional: a complacência e a distensibilidade já se mostraram diferentes (p < 0,001) no período basal sendo maior no grupo NT comparado ao grupo T; mas após a HR 1 e 2, o grupo T mostrou elevação estatísticamente significante destas duas propriedades, sendo que o grupo NT não mostrou tais achados. O diâmetro máximo da artéria braquial mostrou-se aumentado em relação ao basal tanto após HR1 como após HR2 apenas no grupo T. O índice de fluxo total (IFT) não era diferente nos dois grupos no B1 e B2, mas após HR 1 e 2 ambos mostraram aumentos significantes (p < 0,001). Houve correlação positiva entre variáveis CO com PAD e FC no grupo T. No grupo NT não houve correlação do CO com PAS, PAD ou FC. Conclusão O tabagismo altera as propriedades elásticas das artérias e tem papel na hemodinâmica com provável participação do monóxido de carbono.
Objective to evaluate the effects of cigarette smoking on cardiovascular system, with non-invasive methodology, analysing hemodynamic disturbs and arterial elastic properties after being exposed to tobacco. METHODS This study was realized in 45 volunteers, 18 female and 27 male smokers T (n=25, age 40 ± 9 years) and non-smokers NT (n=20, age 39 ± 9 years), who were submitted to the determination of carbon monoxide (CO) to the evaluation of the arterial elastic properties by ultrassonography and test endotelial function (reactive hyperemia - HR) before (B1) and after (B2) the exposition for 20 minutes to a cigarette or a candy. RESULTS Both NT and T groups were similar.The CO concentration in the T group was higher than in the NT group on basal conditions. The CO concentration in the NT group have not changed before and after the proposed exposition (candy) but in the T group it was significant in creased after the cigarette exposition.The systolic blood pressure (PAS) was similar on B1 for both groups, but it was significantly higher on B2 for the T group. The diastolic blood pressure (PAD) on B1 and B2 didn´t exposition significant statistical variation on both groups. The cardiac frequency (FC) presented a different result for both groups with statistical significance ( NT had decrease and T had improved). Regarding the arterial elastic properties and the regional flux response: the complacence and the distensibility were different (p < 0,001) in the basal period and they were higher in the NT group compared to the T group. After HR 1 and 2, the T group showed a significant statistical increase of those two properties and the NT group didn´t show those results. The righest diameter of the brachial artery was increased in relation to the basal diameter after HR1 as well as after HR2 only in the T group. The total flux index (IFT) were not different in both groups in B1 and B2, but after HR1 and 2 both showed significant increase (p < 0,001). There was a positive correlation between the CO variations in relation to PAD and FC in the T group. In the NT group there was a no correlation of CO with PAS, PAD or FC. CONCLUSION Cigarette smoking changes the arterial elastic properties and it has a role in the hemodynamic with possible participation of the carbon monoxide

The Brazilian Unified Public Health System (SUS) shortlisted various plant species of interest (RENISUS) for future clinical use. However, very little is known about their effects on metabolic and transporter proteins, which could potentially lead to herb-drug interactions (HDI). To evaluate this, we conducted in vitro preclinical studies on twenty-four plant extracts to disclose their effects on CYP3A4 mRNA gene expression, intracellular glutathione (GSH) levels, inhibition of γ-glutamyl transferase (GGT) in HepG2 cells and P- glycoprotein (P-gp) activity in vincristine resistant Caco-2 (Caco-2 VCR) cells. We also investigated whether four Brazilian native species were able to activate the human pregnane X receptor (hPXR) in transiently co-transfected HeLa cells. This preclinical research showed that all but two plant extracts were able to modulate at least one of the selected targets. CYP3A4 mRNA gene expression in HepG2 cells was significantly affected by half of the extracts. The antagonistic effect of Solanum paniculatum L. on hPXR could explain its ability to inhibit CYP3A4. GSH levels were affected by 80% of the extracts. There was depletion of intracellular GSH levels by Cordia verbenacea A. DC., Costus spicatus (Jacq.) Sw., Persea americana Mill., Salix alba L., Schinus terebinthifolia Raddi and Syzygium jambolanum (Lam.) DC. accompanied because of the inhibition of GGT activity. P-gp activity was modulated in a significant manner by 17% of the extracts. The approaches used for the conduction of in vitro preclinical studies in herbal medicines revealed a series of challenges faced especially by academics in order to anticipate cases of HDI. Clinicians have also to consider the presence of intrinsic factors such as genetic polymorphisms in each patient. The possible presence of undesirable interactions between RENISUS herbal medicines and essential drugs in SUS need eventually be clinically confirmed to attest our observed in vitro effects.

Resumo: Os confrontos dos animais com situações que induzem medo e ansiedade resultam em uma série de respostas comportamentais defensivas (ex. luta, fuga, imobilidade, vocalização, etc.), ativação neurovegetativa (ex. taquicardia, hipertensão, defecação, etc.), antinocicepção, além de influenciar o comportamento agressivo e aumentar a vulnerabilidade à dependência e recaída ao uso de drogas. Com base no potencial efeito ansiogênico dos neurotransmissores glutamato (via ativação do complexo receptor NMDA-óxido nítrico) e fator liberador de corticotrofina (via receptores CRF1 e CRF2), este estudo investigou o papel desses mediadores, através de injeções sistêmicas, na matéria cinzenta periaquedutal (MCP) ou no núcleo dorsal da rafe (NDR), nas respostas apontadas acima. Os seguintes modelos foram utilizados: labirinto em cruz elevado (LCE, ansiedade), injeção de formalina a 2,5% (nocicepção), conflito intruso-residente (agressão) e estresse de derrota social (dependência à cocaína). Os resultados indicaram: a) o efeito ansiogênico do agonista de receptores NMDA (N-metil-D-aspartato; NMDA) na MCP foi antagonizado pela inibição da enzima de síntese de NO, b) os efeitos ansiogênico e antinociceptivo do CRF na MCP foram via ativação de receptores CRF1 (mas não CRF2) e independentes de NO, c) os efeitos aversivo e antinociceptivo do NO (via administração de um doador de NO) na MCP mostraram-se sensíveis ao bloqueio de receptores CRF1, d) a ativação de receptores CRF2 intra-NDR reduziu o comportamento agressivo induzido pelo conflito intruso-residente, e) o tratamento sistêmico com antagonista CRF1 bloqueou a sensibilização comportamental à cocaína e atenuou o aumento do consumo da mesma induzidos pelo estresse da derrota social
Abstract: Animal confrontation against fear/anxiety-induced situations results in a repertory of behavioral defensive responses (e.g., fight, flight, immobility, vocalization), neurovegetative activation (e.g., tachycardia, hypertension, defecation), antinociception, as well as affects aggressive behavior and increases animals vulnerability to addiction and relapse to drug take. Based on the potential anxiogenic effect elicited by glutamate (via activation of NMDA-nitric oxide receptor complex) and corticotropin releasing factor (via CRF1 and CRF2 receptors), this study investigated the effect of systemic, intra-periaqueductal gray (PAG) or intradorsal raphe nucleus (DRN) injections of these mediators on the above described responses. The following animal models were used: elevated plus maze (EPM, anxiety test), formalin 2.5% injection (nociceptive test), resident-intruder conflict (aggression test) and social defeat stress (to induce cocaine addiction). Results indicated that: a) the anxiogenic effect elicited by intra-PAG injection of glutamate NMDA (N-methyl-D-aspartate; NMDA) receptor agonist was antagonized by prior local infusion an NO synthase inhibitor, b) the anxiogenic and antinociceptive effects elicited by intra-PAG CRF were mediated by CRF1 (but not CRF2) receptor activation and did not depend on NO synthesis, c) the aversive and antinociceptive effects of NO production (induced by intra-PAG injection of a NO donor) were sensitive to CRF1 blockade, d) activation of the CRF2 receptor within the DRN attenuated aggressive behavior elicited by resident-intruder conflict, e) systemic treatment with CRF1 receptor antagonist inhibited cocaine behavioral sensitization and social-defeat stress-induced cocaine consumption
Orientador: Ricardo Luiz Nunes de Souza
Coorientador: Klaus A. Miczek
Banca: Cleopatra da Silva Planeta
Banca: Fabrício Moreira
Banca: Hélio Zangrossi Júnior
Banca: Marcus Lira Brandão
Doutor

A nifedipina atua como bloqueador de canais de cálcio inibindo o fluxo transmembrânico de ions Ca2+ no interior das células do músculo cardíaco e células do músculo liso vascular, o qual induz ao relaxamento do músculo liso e diminuição da resistência vascular periférica. É utilizado no Brasil no tratamento de hipertensão arterial sistêmica (HAS), faz parte da Relação Nacional de Medicamentos Essenciais do Ministério da Saúde e vem sendo distribuída pela Secretaria da Saúde do Estado de São Paulo. Uma vez que a determinação plasmática do fármaco contribui para maior segurança de seu uso, o objetivo deste trabalho se prendeu em padronizar e validar um método analítico em cromatografia em fase gasosa com detecção por captura de elétrons, sensível, específico e reprodutível para a quantificação das concentrações plasmáticas com a finalidade de avaliar a relação entre a dose diária de 60mg e a concentração plasmática versus a resposta da pressão arterial sistêmica, observando a ocorrência de possíveis reações adversas, alterações bioquímicas e hematológicas, em pacientes portadores de HAS, submetidos à farmacoterapia. O método apresentou linearidade na faixa de 10 a 200 ng.mL-1 , com coeficiente de correlação (r) igual a 0,9977. Coeficientes de variação de precisão intra e interensaio menor que 10% e recuperação absoluta da nifedipina de 74,47 a 75,97%. Os limites de detecção e quantificação do método foram de 1,0 e 2,0 ng.mL-1, respectivamente e o fármaco demonstrou ser estável por 90 dias quando armazenado a -70°C ao abrigo da luz. Os dados observados no presente estudo permitiram evidenciar que os pacientes apresentaram concentrações plasmáticas no intervalo terapêutico preconizado, as quais foram efetivas na redução da pressão arterial sistólica e diastólica. Estas concentrações não acarretaram efeitos adversos em nível do sistema hematológico e bioquímico estatisticamente significativos e em relação às reações adversas relacionadas ao medicamento, tais como: cefaléia, edema periférico vascular, tontura, hipotensão arterial, rubor, tosse e cãibras, apesar de relatadas de forma significativa pelos pacientes no inicio do tratamento, foram ao longo do mesmo minimizadas e pouco relatadas.
Nifedipine, a compound of dihydropyridine class, is a calcium-channel antagonists drug that inhibits the transmembrane influx of Ca+2 into cardiac muscle cells and vasculas smooth muscle cells throught specific ion channels. It induces smooth muscle relaxation and decreases peripheral vascular resistance. It is widely used for the treatment of high blood pressure, and is considered as a essencial medicine by the Brazilian government. In Sao Paulo state, this drug has been distributed to hypertensive patients in treatment. Since the drug quantification in plasma contributes for a drug safety use, the objective of the present study was to develop and validate a accurate, specific and reproducible method for the determination of nifedipine in plasma by gas chromatography with eletron capture detection. The validated method was applied in samples of hipertensive patients on 60 mg daily dose of nifedipine with the purpose to evaluate the relation between drug plasma concentration and it\'s daily dose versus the hemodymamic effects, possible side effects and biochemical and hematologic alterations. The method was linear over a concentration range of 10 -200 ng.mL-1 (r2>0,99). The coefficient of variation of intra- and inter-assay precision less than 10% and the recovery was higher than 74%. The limit of detection and quantification were 1,0 and 2,0 ng.mL-1, respectively. Nifedipine was found to be stable in samples stored at -70ºC for 90 days and protect from light. The result showed that patients with drug plasma concentration within therapeutics levels also showed systolic and diastolic blood pressure succesfully controled. Therefore, these patients do not manifested any adverse effects specially in biochemical and hematologic systems. Other adverse efects of nifedipine such as headache, peripheral edema, hypotension, redness, cramp and cough reported by the patients at the beggining of thetreatment, were gradually diminishing and rarely related.

Muito se tem pesquisado sobre os efeitos adversos dos antidepressivos tricíclicos (p.e. imipramina) sobre o sistema respiratório, embora pouco ou quase nada se encontre com relação a tal aspecto na literatura médica sobre a fluoxetina (Prozac®)– um inibidor seletivo da recaptação de serotonina, até porque esta droga começou a ser utilizada somente há cerca de quinze anos. Ambas substâncias (fluoxetina e imipramina, ou seus congêneres) agem basicamente sobre quadros depressivos, fóbicos e obsessivo-compulsivos, obesidade, anorexia, e outras indicações de várias naturezas, indo desde a ansiedade até a sindrome do pânico. Nota-se entretanto, que na maioria dos pacientes com tais quadros (sob tratamento com a fluoxetina ou não), são muito frequentes as queixas de natureza respiratória, como tosse, falta de ar, “angústia" no peito etc. Alguns efeitos adversos da fluoxetina e seus derivados são descritos na literatura, embora raros. Entre eles, destaca-se o comprometimento do aparelho respiratório na forma de doença pulmonar intersticial, pneumonia de hipersensibilidade e fosfolipidose. Apresentamos a seguir um projeto de trabalho experimental em cobaias com intuito de verificar a ação da fluoxetina sobre o aparelho respiratório, com relação à mecânica pulmonar, influência sobre o óxido nítrico exalado e resposta histopatológica pulmonar frente a possíveis injúrias, comparados a animais controles sem efeito da droga. Nosso modelo descreve um protocolo em que os animais foram submetidos ao cloridrato de fluoxetina por via oral durante 30 dias consecutivos, sendo ainda submetidos a uma reação de estresse tipo pânico– a natação forçada.
Although high-affinity imipramine binding sites have been reported in both rat and human lung, the role of the lungs in the pharmacokinetics of antidepressants like fluoxetine has not received much attention. Imipramine and fluoxetine have their action in depression, obesity, panic and anxiety among other indications. However, accumulation of selective serotonin-reuptake inhibitors- like fluoxetine, in the human lungs has been reported. Besides, it is very frequent to most patients (under fluoxetine treatment or not) present respiratory symptoms and/or signs, just like dyspnea, cough, chest anguish and so on. There also have been some adverse effects of fluoxetine described in the literature, although being few and isolated ones, related to hypersensitivity pneumonitis, phospholipidosis and interstitial lung disease. The purpose of this project is to investigate the fluoxetine action in respiratory tract, more specifically over the pulmonary interstitium, using an experimental model in guinea-pigs. Our project comprehends an experimental model with the animals under treatment of fluoxetine so that we could evaluate the substance effects on the respiratory system, concerning the mechanics and the lung histopathological aspects, compared to the control animals. We created then a protocol where the animals were treated with fluoxetine for 30 consecutive days and submmited to the forced swimmig test, a kind of panic-like reaction.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Os confrontos dos animais com situações que induzem medo e ansiedade resultam em uma série de respostas comportamentais defensivas (ex. luta, fuga, imobilidade, vocalização, etc.), ativação neurovegetativa (ex. taquicardia, hipertensão, defecação, etc.), antinocicepção, além de influenciar o comportamento agressivo e aumentar a vulnerabilidade à dependência e recaída ao uso de drogas. Com base no potencial efeito ansiogênico dos neurotransmissores glutamato (via ativação do complexo receptor NMDA-óxido nítrico) e fator liberador de corticotrofina (via receptores CRF1 e CRF2), este estudo investigou o papel desses mediadores, através de injeções sistêmicas, na matéria cinzenta periaquedutal (MCP) ou no núcleo dorsal da rafe (NDR), nas respostas apontadas acima. Os seguintes modelos foram utilizados: labirinto em cruz elevado (LCE, ansiedade), injeção de formalina a 2,5% (nocicepção), conflito intruso-residente (agressão) e estresse de derrota social (dependência à cocaína). Os resultados indicaram: a) o efeito ansiogênico do agonista de receptores NMDA (N-metil-D-aspartato; NMDA) na MCP foi antagonizado pela inibição da enzima de síntese de NO, b) os efeitos ansiogênico e antinociceptivo do CRF na MCP foram via ativação de receptores CRF1 (mas não CRF2) e independentes de NO, c) os efeitos aversivo e antinociceptivo do NO (via administração de um doador de NO) na MCP mostraram-se sensíveis ao bloqueio de receptores CRF1, d) a ativação de receptores CRF2 intra-NDR reduziu o comportamento agressivo induzido pelo conflito intruso-residente, e) o tratamento sistêmico com antagonista CRF1 bloqueou a sensibilização comportamental à cocaína e atenuou o aumento do consumo da mesma induzidos pelo estresse da derrota social
Animal confrontation against fear/anxiety-induced situations results in a repertory of behavioral defensive responses (e.g., fight, flight, immobility, vocalization), neurovegetative activation (e.g., tachycardia, hypertension, defecation), antinociception, as well as affects aggressive behavior and increases animals vulnerability to addiction and relapse to drug take. Based on the potential anxiogenic effect elicited by glutamate (via activation of NMDA-nitric oxide receptor complex) and corticotropin releasing factor (via CRF1 and CRF2 receptors), this study investigated the effect of systemic, intra-periaqueductal gray (PAG) or intradorsal raphe nucleus (DRN) injections of these mediators on the above described responses. The following animal models were used: elevated plus maze (EPM, anxiety test), formalin 2.5% injection (nociceptive test), resident-intruder conflict (aggression test) and social defeat stress (to induce cocaine addiction). Results indicated that: a) the anxiogenic effect elicited by intra-PAG injection of glutamate NMDA (N-methyl-D-aspartate; NMDA) receptor agonist was antagonized by prior local infusion an NO synthase inhibitor, b) the anxiogenic and antinociceptive effects elicited by intra-PAG CRF were mediated by CRF1 (but not CRF2) receptor activation and did not depend on NO synthesis, c) the aversive and antinociceptive effects of NO production (induced by intra-PAG injection of a NO donor) were sensitive to CRF1 blockade, d) activation of the CRF2 receptor within the DRN attenuated aggressive behavior elicited by resident-intruder conflict, e) systemic treatment with CRF1 receptor antagonist inhibited cocaine behavioral sensitization and social-defeat stress-induced cocaine consumption

Syftet är att beskriva symtom och biverkningar som kvinnor med avancerad bröstcancer och cytostatikabehandling rapporterat i ett datoriserat rapporteringssystem före läkarbesök. Undersöka tillfredsställelsen med detta system; se om det finns en skillnad mellan äldre och yngre; undersöka kvinnornas uppfattning om vad som kan förbättras i uppföljningen av symtom/biverkningar samt stödet från läkare. Detta är en kvantitativ, deskriptiv tvärsnittsstudie baserat på rapporteringssystemets databas samt enkätundersökning.

 

Biverkningarna trötthet, smärta och nervpåverkan rapporterades mest frekvent. Tidsåtgången för rapportering ansågs utav de flesta vara kort eller mycket kort och formuläret upplevdes av majoriteten vara ganska lätt till mycket lätt att använda oberoende av datorvana. Läkaren ansågs från hög grad till mycket hög grad vara ett stöd i att hantera symtom och biverkningar av två tredjedelar av respondenterna. Hälften ansåg att rapporterade biverkningar och symtom uppmärksammades av läkaren i hög grad till mycket hög grad.

 

Undersökningen bekräftar det tidigare forskning visat om datoriserade rapporteringssystem i vården, att de är funktionella oavsett ålder samt att intresse finns för att använda dessa i större utsträckning. På grund av litet urval och relativt stort bortfall i enkätstudien kan dock inga direkta slutsatser dras men undersökningen antyder att behov finns att vidareutveckla rapporteringssystemet.

The aim of the study is to describe symptoms and side effects that women with advanced breast cancer and chemotherapy reported in an adverse drug reporting system before seeing their oncologist; examine the satisfaction with this system; if there are any differences between older and younger women; the women’s opinion of what improvements could be done in the follow-up of the symptoms/side effects and the support from the oncologist. This is a quantitative, descriptive cross-sectional study based on the database of the adverse drug reporting system and the questionnaire survey.

 

The side effects fatigue, pain and peripheral neuropathy were most frequently reported. The time consumption for reporting was considered short or very short and the majority thought that the questionnaire was fairly easy to very easy to use independent of computer habits. The oncologists where considered from a high extent to a very high extent being a support in handling the symptoms/side effects by two thirds of the respondents. Two fourths felt that the oncologists attended reported symptoms/side effects from a high extent to a very high extent.

Because of a small sample and a relatively large drop-out no real conclusions can be drawn except the need for further development of the system.

Os efeitos farmacológicos da eritropoetina (EPO) e do tacrolimo (FK 506) têm sido investigados no tratamento da lesão medular, mas são escassos os trabalhos que avaliam a interação entre essas drogas. Neste estudo experimental, 60 ratos Wistar foram submetidos a lesão contusa da medula espinal produzida pelo sistema NYU Impactor. Os animais foram divididos em cinco grupos, sendo: Controle, que recebeu soro fisiológico; EPO, que recebeu eritropoetina; o EPO + FK 506 recebeu EPO associada ao tacrolimo; o FK 506 recebeu tacrolimo. Todas as drogas e o soro fisiológico foram administrados por via intraperitoneal. O grupo Sham foi submetido à lesão medular, mas não recebeu nenhuma droga. Os animais foram avaliados quanto à recuperação da função locomotora em sete diferentes momentos pelo teste de BBB no 2o, 7o, 14o, 21o, 28o, 35o e 42o dias após lesão contusa na medula espinal. No 42o dia, foi realizada avaliação eletrofisiológica dos animais que, logo após, foram sacrificados para análise dos achados histológicos da medula lesionada. Nosso projeto experimental não revelou diferenças na recuperação da função locomotora, nas análises histológica e eletrofisiológica nos animais submetidos ao tratamento farmacológico com eritropoetina e com tacrolimo, após contusão medular torácica
The pharmacological effects of erythropoietin (EPO) and tacrolimus (FK 506) have been investigated in the treatment of spinal cord injuries, but there are few studies that evaluate the interaction between these drugs. In this experimental study, 60 Wistar rats were submitted to contusion spinal cord injury produced by the NYU Impactor system. The animals were divided into five groups: Control, which received saline only; EPO, which received erythropoietin; EPO + FK 506, which received EPO associated with tacrolimus; and the group FK 506, which received tacrolimus. All drugs and saline were administered intraperitoneally. The Sham group underwent spinal cord injury, but did not receive any drug. The animals were evaluated for recovery of locomotor function in seven different times by the BBB test, in the 2nd, 7th, 14th, 21st, 28th, 35th and 42nd days after spinal cord injury. In 42 days, electrophysiological evaluation was performed, and the animals were, shortly after, sacrificed for histological analysis of the injured spinal cord. Our experimental study did not reveal significant differences in the recovery of locomotor function, nor in the histological and electrophysiological analysis in animals treated with erythropoietin and tacrolimus after thoracic spinal cord injury

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Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico
Several pharmaceutical products have been developed in recent years aiming to enhance the treatment of diseases by increasing the effectiveness of drugs. Many of these new products are based on new drug delivery systems. Among these, microemulsions, which were first studied in 1943 by Hoar and Schulman, is of great interest. Microemulsion can be defined as a thermodynamically stable, isotropic, translucent and transparent system of two immiscible liquids stabilized by a surfactant film located in the oil / water interface. The aim os this work was the incorporation of Amphotericin B and Simvasatin to a microemulsion system and analyzes its physicochemical properties and their therapeutical activity when incorporated into this system. Some very promising results were achieved as the reduction of the toxicity and maintenance of the efficacy of the Amphotericin B incorpored into a microemulsion, which was demonstrated in the in vitro pharmacotoxicological study. As for the incorporation of Simvastatin in microemulsion, it was observed a significant improvement in the potential antiinflammatory and anti-infective properties when the system was use to treat infected wounds (simvastatin pleiotropic effects). Therefore, it can be concluded that the incorporation of these drugs into microemulsion system reveal the potential of microemulsions as a promising and novel dosage form, qualifying them for future trials in order to make them available in the pharmaceutical market
In?meros produtos farmac?uticos v?m sendo desenvolvidos nos ?ltimos anos com a finalidade de incrementar o tratamento de doen?as pelo aumento da efic?cia de f?rmacos. Grande parte destes novos produtos est? baseada nos novos sistemas transportadores de f?rmacos. Entre eles destacamse as microemuls?es, que foram primeiramente estudadas em 1943 por Hoar e Schulman. Microemuls?o pode ser definida como um sistema termodinamicamente est?vel, isotr?pico, transl?cido e transparente de dois l?quidos imisc?veis estabilizados por um filme de tensoativos localizados na interface ?leo/?gua. O objetivo deste trabalho foi incorporar anfotericina B e sinvastatina em um sistema microemulsionado e analisar suas propriedades f?sico-qu?micas e suas a??es terap?uticas ap?s a incorpora??o destes f?rmacos ao sistema. Alguns resultados muito promissores foram alcan?ados como a redu??o da toxicidade e a perman?ncia da efic?cia da anfotericina B incorporada em uma microemuls?o durante o estudo farmacotoxicol?gico in vitro. Quanto ? incorpora??o da sinvastatina na microemuls?o, foi constatada uma melhora significativa no potencial antiinflamat?rio e antiinfeccioso (efeitos pleiotr?picos da sinvastatina) em feridas tratadas com esse sistema. Portanto, podemos concluir que a incorpora??o desses f?rmacos em sistemas microemulsionados faz das microemuls?es uma nova e promissora apresenta??o farmac?utica, habilitando-a a futuros ensaios com a finalidade de torn?-los dispon?veis no mercado farmac?utico

A restrição crônica de sal na dieta tem sido recomendada como medida não medicamentosa do tratamento da hipertensão arterial sistêmica. Entre os efeitos observados desta medida terapêutica tem sido descrita uma redução dos valores da pressão arterial (PA), tanto em indivíduos normotensos e pacientes hipertensos, assim como em animais de laboratório. Outros efeitos observados são alterações do metabolismo de carboidratos e de lípides. Tanto em indivíduos normotensos e pacientes hipertensos, assim como em animais de laboratório foram observadas maiores concentrações de peptídeo C e de insulina, sem alteração da glicemia e com redução da captação periférica de glicose pelos tecidos, caracterizando um estado de resistência à insulina. No metabolismo de lípides, uma outra conseqüência da restrição crônica de sal é a maior concentração plasmática de colesterol total e triacilgliceróis. Apesar da demonstração dos efeitos do sal sobre o metabolismo de carboidratos e lípides em humanos e animais existem dados conflitantes na literatura, com resultados opostos a estes descritos. Para melhor compreensão deste fenômeno, foi desenvolvido um estudo em nosso laboratório, o qual demonstrou que ratos Wistar machos que receberam restrição crônica de sal na dieta apresentaram maiores insulinemias medidas durante um teste de tolerância à glicose e sem a constatação de resistência à insulina em adipócitos isolados avaliada pela EC50 Entretanto, o que não ficou esclarecido neste estudo é se este quadro era restrito ao tecido estudado ou se ocorria no animal como um todo, ou seja, se era um fenômeno generalizado. Tendo em vista estes resultados, um outro estudo foi desenvolvido em nosso laboratório. Analisando o animal intacto, foi verificado que a restrição crônica de sal em ratos Wistar estava associada a uma menor sensibilidade à insulina medida por meio de um clamp euglicêmico hiperinsulinêmico (CLAMP) em animais anestesiados. Também foi observado maior peso corpóreo (PC) e maior massa dos tecidos adiposos. No intuito de se compreender quais eram os mecanismos envolvidos na menor sensibilidade à insulina durante a restrição de sal na dieta, também foi feito o pareamento de peso entre os animais que receberam dietas hipo (HO), normo (NR) ou hipersódica (HR) e mesmo sem diferença entre os PC, os animais sob restrição salina mantiveram-se insulino resistentes. Em uma nova etapa foi feito o bloqueio do sistema renina angiotensina com o uso de captopril (inibidor da enzima conversora de angiotensina) ou losartan (antagonista do receptor ATI de angiotensina II) tendo sido verificado que o captopril melhorou a sensibilidade à insulina, o que não ocorreu com o uso de losartan. Outra conseqüência da restrição crônica de sal na dieta é a maior atividade do sistema nervoso simpático (SNS) e uma menor atividade da via L-arginina (LA) / óxido nítrico (NO). Assim, o objetivo deste estudo foi verificar se o SNS e a via LA/NO são mecanismos envolvidos na menor sensibilidade à insulina durante a restrição crônica de sal. Para o desenvolvimento deste estudo, ratos Wistar machos receberam dieta HO, NR ou HR desde o desmame aos 21 dias de vida até completarem 12 semanas. Ao tornarem-se adultos, os ratos foram submetidos ao implante cirúrgico de cateteres e três a cinco dias após era realizado um CLAMP em ratos acordados. No dia do experimento, após um jejum de seis horas se realizava a coleta de sangue e medidas metabólicas e hemodinâmicas. Neste momento, um grupo de ratos recebeu prazosin e propranolol para o bloqueio do SNS e outro grupo recebeu veículo. Um terceiro grupo de animais recebeu LA e outro grupo recebeu D-arginina (DA). Um quinto grupo de animais que consumiu apenas a dieta HR recebeu diltiazem (bloqueador de canal de cálcio). Quarenta e cinco minutos após a infusão das drogas o CLAMP foi iniciado. Foram medidos PC, glicemias, insulinemias, lípides, nitrato/nitrito (NOx), PA sistólica (PAS), diastólica (PAD) e freqüência cardíaca (FC). O PC foi maior na dieta HO do que na NR e HR. Também foi observado maior PC na dieta NR do que HR. Em situação basal observaram-se maiores valores de glicemia e insulinemia durante a restrição salina em comparação aos ratos em dieta NR e HR. A PAS e PAD foram maiores na dieta HR em comparação aos ratos em dieta NR e HO, enquanto a FC foi maior nos ratos em dieta HO em comparação àqueles sob dietas NR e HR. Também foram observadas maiores concentrações plasmáticas de colesterol total (COL) e triacilgliceróis (TAG) durante a restrição salina. Os animais controle apresentaram menor sensibilidade à insulina em comparação àqueles em dieta NR ou HR. O bloqueio do SNS corrigiu o efeito do sal sobre a captação de glicose, não sendo mais observada diferença entre as dietas. O bloqueio do SNS não influenciou a glicemia mas reduziu a maior insulinemia nos ratos em dieta HO, não sendo mais observada diferença entre as dietas quanto às insulinemias ao início do CLAMP. A infusão de LA melhorou a menor captação periférica de glicose nos ratos em dieta HO, sem influência sobre os animais tratados com dieta NR ou HR. Diltiazen não modificou a sensibilidade à insulina apesar de ter reduzido a PA em intensidade semelhante ao bloqueio do SNS. O bloqueio do SNS reduziu os valores de PAS e PAD nas três dietas, com queda mais intensa dos seus valores nos animais sob sobrecarga salina, logo ao início do CLAMP. Diltiazen reduziu a PAS e PAD. DA não influenciou (p>0,05) os valores de PAS e PAD. Já a infusão de LA promoveu redução dos valores de PAS e PAD somente em ratos em sobrecarga salina e reduziu os maiores valores de FC observados nos ratos durante a restrição de sal na dieta. O bloqueio do SNS e a infusão de LA reduziram os maiores valores de TAG ao término do CLAMP, o que não foi observado nos grupos veículo e DA. Não foi observada modificação da concentração plasmática de COL entre os grupos independentemente dos fármacos ministrados. Durante a restrição salina foi observado menor concentração plasmática de NOx em comparação à sobrecarga salina. A infusão de LA promoveu um incremento na concentração plasmática de NOx ao término do CLAMP nas três dietas, o que não foi observado nos ratos tratados com DA. Ao término do CLAMP não foram observadas diferenças na concentração de NOx plasmático entre as dietas nos animais que receberam LA. A concentração plasmática de NOx nas três dietas foi menor nos ratos que receberam DA. Assim, o bloqueio do SNS e a ativação da via LA/NO melhoraram os efeitos metabólicos decorrentes da restrição crônica de sal.
Chronic dietary salt restriction is recommended as non-pharmacological measure of hypertension treatment. One of the observed effects of this therapeutic measure is the blood pressure (BP) decrease in normotensive subjects and in hypertensive patients, and also in laboratory animals. Another side effects observed are disorders of carbohydrate and lipid metabolism. Salt restriction induced higher C-peptide and insulin levels, without changes on plasma glucose, and lower glucose uptake by the tissues in normotensive subjects and in hypertensive patients, and also in laboratory animals, suggesting an insulin resistant state. Other consequences of salt restriction are higher plasma levels of cholesterol and triacylglycerols. Some studies have disclosed opposite results. With the objective to better understand these phenomena, a study was developed in our laboratory, that showed that male Wistar rats on chronic salt restriction presented higher insulin plasma levels measured during a glucose tolerance test, without insulin resistance in isolated adipocytes measured by the EC50 of the insulin - glucose uptake curve. It was not clear in this study if this phenomenon was restricted to the evaluated tissue or if it was a phenomenon in the whole animal. With these results, another study was developed in our laboratory. Analyzing the whole animal, chronic salt restriction in male Wistar rats was associated with lower insulin sensitivity measured by a euglycemic hyperinsulinemic clamp (CLAMP) in anesthetized rats. Higher body weight (BW) and adipose tissue mass was also observed. With the objective to understand the involved mechanisms in the lower insulin sensitivity due to dietary salt restriction, weight was paired among animals on low (LSD), normal (NSD) or high (HSD) salt diet, and even without BW difference, salt restricted animals were still insulin resistants. In another step, renin angiotensin system blockade with captopril (angiotensin enzyme conversion inhibitor) or losartan (angiotensin II type I receptor antagonist) was performed. It was observed that captopril, but not losartan, improved insulin sensitivity. Another consequence of salt restriction is a higher sympathetic nervous system (SNS) and a lower L-arginine (LA) / nitric oxide (NO) pathway activity. The objective of this study was to verify if SNS and LA / NO are mechanisms involved in the lower insulin sensitivity due to chronic salt restriction. Male Wistar rats received LSD, NSD, or HSD since weaning until adulthood. In the 12th week of age, catheters were inserted and three to five days latter, a CLAMP was performed in awaked rats. On the day of the experiment, after six hours fasting, blood samples were collected and metabolic and hemodynamic measures were done. At this moment, a group of rats received prazosin and propranolol for SNS blockade and another group received vehicle. A third group of animals received LA and a fourth group received D-arginine (DA). Another group of rats only on HSD received diltiazen (calcium channel blocker). Fourty five minutes after drug infusion the CLAMP was started. BW, plasma glucose, insulin, lipids, and nitrate/nitrite (NOx), systolic BP (SBP), diastolic (DBP), and heart rate (HR) were measured. BW was higher on LSD than on NSD and HSD. BW was also higher on NSD than HSD. Basal plasma glucose and insulin were higher during salt restriction than on NSD and HSD. SBP and DBP were higher on HSD than on NSD and LSD, and HR was higher on LSD than on NSD and HSD. Cholesterol (CHOL) and triacylglycerol (TAG) plasma levels were higher on salt restriction. LSD rats presented lower insulin sensitivity compared to animals on NSD or HSD. SNS blockade corrected effect of salt on glucose uptake. SNS blockade had no influence on glucose levels but reduced the higher plasma insulin in LSD rats, without differences in insulin levels between diets at the start of the CLAMP. LA improved the lower glucose uptake observed in LSD rats, with no influence on the rats on NSD or HSD. Diltiazen had no effect on insulin sensitivity. SNS blockade reduced SBP and DBP in rats on the three diets, with an intense BP fall on HSD rats at the start of the CLAMP. Diltiazen reduced SBP and DBP. DA had no influence in SBP and DBP. On the other hand, LA decreased SBP and DBP only in salt overloaded rats and reduced the higher HR observed on salt restricted rats. SNS blockade and LA infusion reduced the higher TAG concentration at the end of the CLAMP, which was not observed in vehicle and DA groups. COL level was not influenced by drug infusion. During salt restriction, lower plasma NOx was observed compared to salt overload. LA infusion promoted plasma NOx increment at the end of the CLAMP. At the end of the CLAMP, no difference was observed in plasma NOx among the rats on the three salt diets and infused with LA. Plasma NOx was lower in rats in the DA group. In conclusion, SNS blockade and LA/NO pathway activation improved the metabolic effects due to chronic dietary salt restriction.

碩士
中國醫藥大學
營養學系碩士班
102
In general, people exposed to a wide variety of environmental pollutants, toxicants, drugs, and food additives that may cause toxicity. To minimize chemical insults, converting these substances into hydrophilic metabolites by drug-metabolizing system is necessary. Shikonin, a naphthoquinone pigment, is rich in the root of Chinese herbal plant Lithospermum erythrorhizon. Evidence indicates that shikonin displays multiple physiological functions including anti-microbes, anti-inflammation, and anti-oxidation. In this study, we intend to examine the effect of shikonin on hepatic Phase I and Phase II drug-metabolizing enzymes as well as Phase III membrane transport proteins expression and the possible mechanism involved. Twenty-four h post plating, rat primary hepatocytes were treated with 0-2 μM shikonin for 16 or 24 h. Immunoblotting assay and RT-PCR revealed that shikonin dose-dependently increases cytochrome P450 (CYP) 1A1/2, CYP3A2, CYP2D1, and CYP2C6) and pi class of glutathione S-transferase (PGST), UDP glucuronosyltransferase 1A1 (UGT1A1), and NADP(H) quinone oxidoreductase 1 (NQO1) protein and RNA expression. Moreover, the protein and mRNA levels of Phase III membrane transport proteins organic anion-transporting polypeptide (OATP) 1B1, OATP2B1, P-glycoprotein, and multidrug resistance-associated protein (MRP) 2/3 were also increased by shikonin. In addition, nuclear translocation of aryl hydrocarbon receptor (AhR) and NF-E2-related factor 2 (Nrf2) was augmented by shikonin. EMSA confirmed that shikonin increases AhR, pregnane X receptor, and Nrf2 binding to dioxin-responsive element, direct repeat 4 (DR4), and antioxidant-responsive element, respectively.Shikonin time-dependently increased ERK1/2, JNK, and p38 phosphorylation up to 30 min. ERK inhibitor (PD98059), JNK inhibitor (SP600125), and p38 inhibitor (SB2035801) pretreatment suppressed shikonin-induced Nrf2 nuclear translocation. Moreover, AhR and Nrf2 knockdown attenuated shikonin-induced increase in CYP1A1/2 induction and NQO1 and PGST expression, respectively. Taken together, results suggest that shikonin effectively upregulates the transcription of Phase I and II drug-metabolizing enzyme and Phase III membrane transporters at least partially through activation of AhR, PXR, and Nrf2 and Nrf2-dependent activation is MAPK-dependent.

Lymph is primarily composed of fluid and proteins from the blood circulatory system that drain into the space surrounding cells, interstitial space. From the interstitial space, the fluid enters and circulates in the lymphatic system until it is delivered into the venous system. In contrast to the blood circulatory system, the lymphatic system lacks a central pumping organ dictating the predominant driving pressure and velocity of lymph. Transport of lymph via capillaries, pre-collecting and collecting lymphatic vessels relies on the synergy between pressure gradients, local tissue motion, valves and lymphatic vessel contractility. The direction of lymph transport is regulated by bicuspid valves distributed throughout pre-collecting and collecting lymphatic vessels. Effective transport of lymph into the venous system is of prime importance. Disruption of lymph transport, because of impaired lymphatic function, reduced numbers of vessels or valvular insufficiencies can have severe health consequences, including lymphedema for which current clinical therapies are not curative. The lymphatic valves are usually bicuspid, however, congenital malformations in the valve such as single leaflet valve formation and arrested lymphatic valve development are observed and can cause lymphedema. Here we employ 4-week-old mice to study the effects of valves and malformed valves on lymph transport shedding light into some of the potentially underlying consequences of lymphedema. Polyethylene glycol (PEG) coated latex particles were injected into the inguinal lymph node of anesthetized mice. Particle displacement measurements through efferent lymphatic vessels yielded velocity, wall shear stress, vorticity and strain of the efferent lymph flow field carrying lymph from subdermal inguinal lymph nodes. Lymphatic vessel endothelial Prox1 green fluorescent protein (GFP) marker enabled the detection of lymphatic vessel walls and valves. Flow field, flow velocity, flow rate, velocity profiles, wall shear stress, vorticity and strain values were compared in regions downstream of normal and malformed valves in two wild type mice. A Clec2-deficient mouse, which experiences lymphatic development defects and is used as a lymphedema model, was employed to further elucidate the lymphatic valves on transport. The absence of centralized pumping yields highly variable lymphatic flow cycles varying from one to fifteen seconds. The presence of lymphatic valves introduces boundary conditions that yield spatial and temporal flow gradients increasing the degree of complexity of lymph transport. The valves dictate the trajectory of the particles and promote the formation of recirculation zones. Even in the presence of valves, lymph flow commonly reverses. Congenital defects like a single leaflet valve lowers the lymph flow efficiency and promotes higher wall shear stress regions. Furthermore, the absence of functional valves in the Clec2-deficient mouse not displaying lymphedema yielded lymph flow lacking the pulsatility that characterizes normal lymphatic flow.

碩士
國立中央大學
網路學習科技研究所
103
The spread of drugs causes serious problems recently. To enhance people awareness of drugs’ harm, several approaches were adopted to deliver anti-drug materials. However, limitations were observed in these approaches. Thus, this research develops a game-based anti-drug system (GADS), which incorporates anti-drug materials into game-based learning. However, previous research indicated that game-based learning may cause some problems, such as the increase of learners’ cognitive overload. Nevertheless, not all of learners can overcome these problems because learners have diverse characteristics. Thus, two approaches, i.e., customization and personalization, can be applied to accommodate students’ individual differences, especially drug addiction and prior game knowledge. However, such two approaches have different advantages and disadvantages. Thus, there is a need to compare the effectiveness of these two approaches. In order to get a complete understanding of whether the customized and personalized GADS are suitable to learners with different levels of drug addiction and prior game knowledge, two empirical studies were conducted. In Study One, a customized game-based anti-drug system (CGADS) is developed and drug experience and prior game knowledge were considered as a target to investigate learners’ reactions to the CGADS. The results indicated that non-addictive learners had better game performance and learning performance than addictive learners Moreover, non-addictive learners focused on playing digital games while addictive learners focused on learning anti-drug materials during the gaming process. On the other hand, learners with high prior game knowledge showed better game performance and they thought that the CGADS was too simple while learners with low prior game knowledge frequently completed the game quests and they did not favor the game hints provided by the CGADS. Based on such different preferences, a personalized game-based anti-drug system (PGADS) was developed to further examine how drug addiction and prior game knowledge affected learners’ reactions to the CGBLS and PGBLS differently in Study Two. The results showed that learners with the PGADS had better learning performance and perceptions than those with the CGADS. Moreover, learners with the PGADS learned the anti-drug materials completely. Based on the results of each study, a framework is produced. This framework can be applied to support researchers, learners and designers to promote the effectiveness of game-based anti-drug systems.

Research Doctorate - Doctor of Philosophy (PhD)
Self-poisoning with pharmaceutical agents is very common across the world. Central nervous system depressant drugs (CNS-Ds) are among the most common substances taken in overdose in hospital-treated episodes of self-poisoning in Australia, the UK and the US. The majority of the patients with CNS-D overdose treated in hospitals in Australia and the UK are discharged within 24-48 hours of their admission, when they still could potentially have subclinical effects of those drugs. This thesis systematically reviews published evidence on the effects of CNS-Ds on cognitive functions (Chapter 2), automobile driving and traffic accidents (Chapter 3, Paper 1), and presents original research conducted to examine the effects of CNS-D overdose on cognitive functions underpinning daily activities (Chapter 5, Paper 2), surrogate bedside tests of driving skills (Chapter 6, Paper 3) and risk of traffic accidents (Chapter 7, Paper 4) of patients discharged from hospital following treatment. Comprehensive neuropsychological assessment shows that patients discharged after treatment for CNS-D overdose have significant residual impairments in multiple cognitive functions including visual attention and visuomotor skills, decision-making, and executive functions and working memory (Chapter 5). The impairments, as estimated by regression models, were equivalent to a ‘cognitive ageing’ of 10-20 years depending on the domain tested. Furthermore, executive dysfunction of the patients tends to worsen with increasing task demands. Converging evidence from the neuropsychological assessment and epidemiological approach indicates that CNS-D overdose has deleterious effects on driving. In particular, the performance of Trail-Making Test B, when interpreted with respect to its correlation with driving performance and traffic accident risk, suggests that nearly two-thirds of the patients with CNS-D overdose may be grossly impaired (equal or less than 10th percentile) at the time of discharge from hospital (Chapter 6, Paper 3). The epidemiological evidence (Chapter 7, Paper 4) shows that the traffic accident risk of these individuals increases by 3-4 times in the immediate post-discharge period, and remains nearly twice their baseline risk after one week following overdose. In the concluding chapter (Chapter 8), we examine the impact of these impairments on daily activities that the discharged patients are expected and likely to carryout during the post-discharge period, and discuss the clinical implications in post-discharge management of patients treated for CNS-D overdose.

by Lam Heung-wah, Angora.
Thesis (M. Phil.)--Chinese University of Hong Kong, 1992.
Includes bibliographical references (leaves 124-131).
ACKNOWLEDGEMENTS --- p.iv
ABSTRACT --- p.v
LIST OF ABBREVIATIONS --- p.vii
LIST OF TABLES --- p.viii
LIST OF FIGURES --- p.ix
INTRODUCTION --- p.1
LITERATURE REVIEW --- p.4
Chapter A. --- Some Aspects of Cardiovascular Physiology and Pharmacology --- p.4
Chapter I. --- Fundamental Principles Governing regulation of Arterial Pressure --- p.4
Chapter II. --- Hypertension --- p.12
Chapter III. --- Antihypertensive Substances --- p.16
Chapter B. --- Mushrooms and Their Medicinal Values --- p.29
Chapter I. --- "The Straw Mushroom, V. volvacea" --- p.29
Chapter II. --- "Mushrooms, Blood Pressure, and Related Changes" --- p.32
MATERIALS AND METHODS --- p.39
Chapter A. --- Basic Preparative Procedures --- p.39
Chapter I. --- Preparation of Straw Mushroom Extract (SME) --- p.39
Chapter II. --- Purification of SME by Dialysis --- p.40
Chapter III. --- Preparation for In vivo Blood Pressure Measurement in Rats --- p.40
Chapter IV. --- preparation of Right Atrium for In vitro Studies --- p.41
Chapter V. --- Preparation of Artery Strip for In vitro Studies --- p.42
Chapter B. --- Experiments Done --- p.43
Chapter Experiment 1. --- Toxicity of SME --- p.43
Chapter Experiment 2. --- Hypotensive Effect of SME and Dialyzed Samples --- p.44
Chapter Experiment 3. --- Pharmacological Antagonist Studies --- p.45
Chapter Experiment 4. --- Effect of Autonomic Ganglion Blocker and Alpha Blocker on Hypotensive Changes Induced by SME --- p.47
Chapter Experiment 5. --- Study on Renin-Angiotensin and Kinin Systems --- p.48
Chapter Experiment 6. --- Urinary and Sodium Excretion in Water-loaded Rats --- p.48
Chapter Experiment 7. --- Chronotropic and Inotropic Studies on Isolated Right Atria --- p.49
Chapter Experiment 8. --- Contractile Responses of SME & Its Dialyzed Samples on Rat Tail Artery Strips --- p.50
Chapter Experiment 9. --- Effect of Adrenergic Blockers in SME Preconstricted Strips --- p.51
Chapter Experiment 10. --- Acute Oral Effect of DUL8000 and DLL8000 from SME on Blood Pressure --- p.51
Chapter Experiment 11. --- "Chronic Oral Effect of SME on Blood Pressure, Total Free Cholesterol and Triglyceride Levels" --- p.52
Chapter C. --- Statistics --- p.55
RESULTS --- p.56
Chapter A. --- Toxicity of Straw Mushroom Extract (SME) --- p.56
Chapter B. --- Effects of SME in Normotensive Rats --- p.56
Chapter I. --- Blood Pressure Changes --- p.56
Chapter II. --- Pharmacological Antagonists Studies --- p.58
Chapter III. --- Converting Enzyme Activity --- p.60
Chapter IV. --- Urinary and Sodium Excretion --- p.60
Chapter V. --- In vitro Arterial and Cardiac Effects --- p.61
Chapter C. --- Cardiovascular Effects of Dialyzed Samples of SME (Molecular Mass cutoffl2000) in Spontaneously Hypertensive Rats (SHR) and Normotensive Rats --- p.61
Chapter I. --- Blood Pressure Changes --- p.61
Chapter II. --- In vitro Arterial and Cardiac Effects --- p.62
Chapter D. --- Cardiovascular Effects of Dialyzed Samples (DUL8000 and DLL8000) in Spontaneously Hypertensive Rats (SHR) and Normotensive Rats --- p.63
Chapter I. --- Blood Pressure Changes --- p.63
Chapter II. --- In vitro Arterial and Cardiac Effects --- p.65
Chapter E. --- The Acute Oral Effect of SME on Blood Pressure --- p.68
Chapter F. --- "Chronic Dietary Effect of SME on Blood Pressure, Total Free Serum Cholesterol and Triglyceride Levels" --- p.68
DISCUSSION --- p.109
Chapter A. --- The Hypotensive Effect of SME --- p.109
Chapter B. --- The Hypotensive Action of SME: Mechanism of Action --- p.110
Chapter C. --- The Cardiovascular Active Fractions in SME --- p.113
Chapter D. --- "The Cardiovascular Effect of DUL8000 and DLL8000 in Rats with Reference to Age, Sex and Strains" --- p.115
Chapter E. --- The Oral Effect of SME on Blood Pressure and on Serum Cholesterol and Triglyceride Levels --- p.118
SUMMARY --- p.121
REFERENCES --- p.124
APPENDIXES --- p.132

Yang, Chao.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2011.
Includes bibliographical references (leaves 137-146).
Abstracts in English and Chinese.
Thesis Committee --- p.i
Acknowledgements --- p.ii
Contents --- p.iii
Declaration --- p.vii
Abstract --- p.viii
摘要 --- p.xi
Abbreviations --- p.xiii
Chapter CHAPTER ONE: --- INTRODUCTION
Chapter 1.1 --- Occurrence and Structure of Phytosterols in Plants --- p.P.1
Chapter 1.2 --- Biological Effects of Phytosterols
Chapter 1.2.1 --- Cholesterol-lowering Effect of Phytosterols --- p.P.3
Chapter 1.2.2 --- Anti-cancer Effect of Phytosterols --- p.P.5
Chapter 1.2.3 --- Anti-proliferative Effect of Phytosterols --- p.P.5
Chapter 1.3 --- Intake and Absorption of Phytosterols in Human Beings --- p.P.6
Chapter 1.4 --- Occurrence and Physiological Levels of Phytosterol Oxidation Products (POPs)
Chapter 1.4.1 --- Occurrence of POPs --- p.P.8
Chapter 1.4.2 --- Physiological Levels of POPs --- p.P.8
Chapter 1.5 --- Endothelium and the Vascular Tone
Chapter 1.5.1 --- Role of Endothelium in the Control of Vascular Tone --- p.P.11
Chapter 1.5.2 --- "Endothelial Dysfunction, Cholesterol Oxidation Products (COPs) and Phytosterol Oxidation Products (POPs)" --- p.P.12
Chapter 1.6 --- Calcium Homeostasis in the Vascular Smooth Muscle Cells (VSMCs)
Chapter 1.6.1 --- Modes of Ca2+ Entry in VSMCs --- p.P.15
Chapter 1.6.2 --- Modes of Ca2+ Efflux in VSMCs --- p.P.18
Chapter 1.7 --- Objectives of the Study --- p.P.19
Chapter CHAPTER TWO: --- β-SITOSTEROL OXIDATION PRODUCTS ATTENUATE VASORELAXATION BY INCREASING REACTIVE OXYGEN SPECIES AND CYCLOOXYGENASE-2
Chapter 2.1 --- Introduction --- p.P.21
Chapter 2.2 --- Materials and Methods
Chapter 2.2.1 --- Preparation of SOPs --- p.P.24
Chapter 2.2.2 --- Gas Chromatography -mass Spectrometry (GC-MS) Identification of SOPs --- p.P.24
Chapter 2.2.3 --- Analysis of SOPs --- p.P.25
Chapter 2.2.4 --- Vessel Preparation --- p.P.25
Chapter 2.2.5 --- Isometric Force Measurement --- p.P.26
Chapter 2.2.6 --- Western Blotting --- p.P.27
Chapter 2.2.7 --- Primary Culture of Rat Aortic Endothelial Cell --- p.P.28
Chapter 2.2.8 --- Measurement of SOPs-induced Intracellular Oxidative Stress --- p.P.29
Chapter 2.2.9 --- Drugs --- p.P.30
Chapter 2.2.10 --- Data Analysis --- p.P.30
Chapter 2.3 --- Results
Chapter 2.3.1 --- GC-MS Identification of SOPs --- p.P.32
Chapter 2.3.2 --- Analysis of SOPs --- p.P.34
Chapter 2.3.3 --- SOPs But Not β-Sitosterol Impaired ACh- and A23187-induced relaxations --- p.P.36
Chapter 2.3.4 --- Inhibition of COX Pathway Reversed SOPs-induced Impairment in Relaxation --- p.P.39
Chapter 2.3.5 --- SOPs Elevated Endothelial COX-2 Expression --- p.P.42
Chapter 2.3.6 --- SOPs Increased COX-2 Expression via An Oxidative Stress-sensitive Pathway --- p.P.45
Chapter 2.4 --- Discussion --- p.P.52
Chapter 2.5 --- Conclusion --- p.P.56
Chapter CHAPTER THREE: --- β-SITOSTEROL OXIDATION PRODUCTS POSSESS POTENTIAL VOCC BLOCKING EFFECT IN VSMCs
Chapter 3.1 --- Introduction
Chapter 3.1.1 --- 2+ Modes of Ca Entry and Efflux in Vascular Smooth Muscle Cells (VSMCs) --- p.P.57
Chapter 3.1.2 --- Effect of Cholesterol and COPs on VSMCs --- p.P.57
Chapter 3.2 --- Methodology and Materials
Chapter 3.2.1 --- Vessel Preparation --- p.P.59
Chapter 3.2.2 --- Isometric Force Measurement iv --- p.P.59
Chapter 3.2.3 --- Drugs --- p.P.60
Chapter 3.2.4 --- Data Analysis --- p.P.61
Chapter 3.3 --- Results
Chapter 3.3.1 --- SOPs but not β-Sitosterol Induced Relaxation in 60 mM K+ -preconstricted Endothelium-denuded Aorta --- p.P.62
Chapter 3.3.2 --- Both SOPs and β-Sitosterol did not Relax U46619-preconstricted Endothelium-denuded Aorta --- p.P.64
Chapter 3.3.3 --- Both SOPs and β-Sitosterol did not Relax PDA -preconstricted Endothelium-denuded Aorta --- p.P.66
Chapter 3.3.4 --- SOPs Attenuated 60 mM K+-induced Contraction --- p.P.68
Chapter 3.3.5 --- SOPs Attenuated Phenylephrine-induced Contraction --- p.P.70
Chapter 3.3.6 --- Effect of SOPs on Concentration-dependent Responses to U46619 --- p.P.72
Chapter 3.3.7 --- Preincubation with Bay K 8644 Abolished SOPs-induced Relaxation in 60 mM K+ -preconstricted Rings --- p.P.74
Chapter 3.3.8 --- Preincubation with Thapsigargin did not Affect SOPs-induced Relaxation in 60 mM K+ -preconstricted Rings --- p.P.76
Chapter 3.3.9 --- Preincubation with Ouabain did not Affect SOPs-induced Relaxation in 60 mM K+ -preconstricted Rings --- p.P.78
Chapter 3.3.10 --- Preincubation with Nickel Potentiated SOPs-induced Relaxation in 60 mM K+ -preconstricted Rings --- p.P.80
Chapter 3.4 --- Discussion --- p.P.84
Chapter 3.5 --- Conclusion and Future Work --- p.P.88
Chapter CHAPTER FOUR: --- INVOLEMENT OF NITRIC OXIDE IN THE PROTECTIVE EFFECTS OF PHYTOSTEROLS AGAINST HOMOCYSTEINE-INDUCED IMPAIRMENT OF ENDOTHELIUM-DEPENDENT RELAXATIONS OF RAT AORTA
Chapter 4.1 --- Introduction --- p.P.89
Chapter 4.2 --- Materials and Method
Chapter 4.2.1 --- Vessel Preparation --- p.P.93
Chapter 4.2.2 --- Isometric Force Measurement --- p.P.93
Chapter 4.2.3 --- Western Blotting --- p.P.94
Chapter 4.2.4 --- "1,1 -diphenyl-2-picrylhydrazyl (DPPH) Radical Scavenging Capacity" --- p.P.96
Chapter 4.2.5 --- Primary Culture of Rat Aortic Endothelial Cells V --- p.P.96
Chapter 4.2.6 --- Measurement Intracellular Oxidative Stress --- p.P.97
Chapter 4.2.7 --- Nitric Oxide (NO) Measurement --- p.P.97
Chapter 4.2.8 --- Drugs --- p.P.98
Chapter 4.2.9 --- Data Analysis --- p.P.99
Chapter 4.3 --- Results
Chapter 4.3.1 --- Impairment of Endothelium-dependent Relaxation by HC was Reversed by ROS Scavenger --- p.P.100
Chapter 4.3.2 --- Brassicasterol Reversed HC-induced Endothelial Dysfunction In a Dose-dependent Manner --- p.P.102
Chapter 4.3.3 --- β-Sitosterol and Stigmasterol Reversed HC-induced Endothelial Dysfunction --- p.P.104
Chapter 4.3.4 --- Effects of β-Sitosterol Oxidation Products (SOPs) on HC-induced Endothelial Dysfunction --- p.P.106
Chapter 4.3.5 --- Effects of Brassicasterol and β-Sitosterol on H2O2-induced Impairment of Endothelium-dependent Relaxation --- p.P.108
Chapter 4.3.6 --- Phytosterols did not Directly Scavenge Free Radicals --- p.P.110
Chapter 4.3.7 --- "HC and Brassicasterol did not Affect the Expression of SOD-1, SOD-2, eNOS, COX-1 and COX-2 in Aorta" --- p.P.112
Chapter 4.3.8 --- HC Increased ROS Production in Primary Rat Aortic Endotelial Cells --- p.P.116
Chapter 4.3.9 --- Brassicasterol did not Reverse the ROS Production by HC treatment In the Endothelial Cells --- p.P.120
Chapter 4.3.10 --- Effect of L-NAME on Reversing the Effect of Brassicasterol on ACh-induced Relaxation --- p.P.123
Chapter 4.3.11 --- Brassicasterol Reversed the Inhibitory Effect of HC on ACh-induced NO Production in Endothelial Cells --- p.P.125
Chapter 4.4 --- Discussion --- p.P.128
Chapter 4.5 --- Conclusion and Future Work --- p.P.132
Chapter CHAPTER FIVE: --- CONCLUSIONS AND FUTURE WORK --- p.P.134
Chapter CHAPTER SIX: --- REFERENCES --- p.P.137

Indiana University-Purdue University Indianapolis (IUPUI)
Traditional treatment strategy development for diseases involves the identification of target proteins related to disease states, and the interference of these proteins with drug molecules. Computational drug discovery and virtual screening from thousands of chemical compounds have accelerated this process. The thesis presents a comprehensive framework of computational drug discovery using system biology approaches. The thesis mainly consists of two parts: disease biomarker identification and disease treatment discoveries. The first part of the thesis focuses on the research in biomarker identification for human diseases in the post-genomic era with an emphasis in system biology approaches such as using the protein interaction networks. There are two major types of biomarkers: Diagnostic Biomarker is expected to detect a given type of disease in an individual with both high sensitivity and specificity; Predictive Biomarker serves to predict drug response before treatment is started. Both are essential before we even start seeking any treatment for the patients. In this part, we first studied how the coverage of the disease genes, the protein interaction quality, and gene ranking strategies can affect the identification of disease genes. Second, we addressed the challenge of constructing a central database to collect the system level data such as protein interaction, pathway, etc. Finally, we built case studies for biomarker identification for using dabetes as a case study. The second part of the thesis mainly addresses how to find treatments after disease identification. It specifically focuses on computational drug repositioning due to its low lost, few translational issues and other benefits. First, we described how to implement literature mining approaches to build the disease-protein-drug connectivity map and demonstrated its superior performances compared to other existing applications. Second, we presented a valuable drug-protein directionality database which filled the research gap of lacking alternatives for the experimental CMAP in computational drug discovery field. We also extended the correlation based ranking algorithms by including the underlying topology among proteins. Finally, we demonstrated how to study drug repositioning beyond genomic level and from one dimension to two dimensions with clinical side effect as prediction features.

Ng Chi Ho.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2005.
Includes bibliographical references (leaves 147-160).
Abstracts in English and Chinese.
ACKNOWLEDGMENTS --- p.I
ABSTRACT --- p.II
LIST OF ABBREVIATIONS --- p.VII
TABLE OF CONTENTS --- p.IX
Chapter CHAPTER 1 --- GENERAL INTRODUCTION
Chapter 1.1 --- Introduction of Low-density lipoprotein --- p.1
Chapter 1.1.1 --- What are lipids? --- p.1
Chapter 1.1.2 --- Function and structure of cholesterol --- p.1
Chapter 1.1.3 --- Function and classification of lipoprotein --- p.1
Chapter 1.2 --- Functions of low-density lipoprotein --- p.2
Chapter 1.3 --- Basic structure of low-density lipoprotein --- p.4
Chapter 1.4 --- Principle on isolation and purification of low-density lipoprotein --- p.4
Chapter 1.5 --- Cholesterol transport system --- p.7
Chapter 1.5.1 --- Exogenous pathway of cholesterol metabolism --- p.7
Chapter 1.5.2 --- Endogenous pathway of cholesterol metabolism --- p.7
Chapter 1.5.3 --- Reverse transport of Cholesterol --- p.8
Chapter 1.6 --- Oxidation of LDL --- p.10
Chapter 1.6.1 --- Agents that causes oxidation --- p.10
Chapter 1.6.1.1 --- Lipoxygenases --- p.10
Chapter 1.6.1.2 --- Myeloperoxidase --- p.10
Chapter 1.6.1.3 --- Reactive nitrogen species --- p.11
Chapter 1.6.1.4 --- Reactive oxygen species --- p.11
Chapter 1.6.2 --- Factors that affect the susceptibility of LDL oxidation --- p.13
Chapter 1.7 --- Hyperlipidaemia 一 chance to increase LDL oxidation --- p.13
Chapter 1.7.1 --- Definition of hyperlipidemia and hypercholesterolemia --- p.13
Chapter 1.7.2 --- Risk factors of hyperlipidaemia --- p.13
Chapter 1.7.2.1 --- High fat low fibre diets: --- p.13
Chapter 1.7.2.2 --- Obesity --- p.14
Chapter 1.7.2.3 --- Type II diabetes --- p.14
Chapter 1.7.2.4 --- Genetic factors (Familial hyperlipidemias) --- p.14
Chapter 1.8 --- Diseases related to oxidized LDL --- p.15
Chapter 1.8.1 --- Cardiovascular diseases --- p.15
Chapter 1.8.1.1 --- Atherosclerosis and ischemic heart attack --- p.15
Chapter 1.8.1.2 --- Factors that affect incidence of atherosclerosis --- p.16
Chapter 1.8.1.2.1 --- Triglyceride-rich lipoprotein --- p.16
Chapter 1.8.1.2.2 --- Small and dense LDL --- p.16
Chapter 1.8.1.3 --- Stroke --- p.17
Chapter 1.8.2 --- Common ways to reduce plasma cholesterol level --- p.17
Chapter 1.8.2.1 --- Diet control --- p.17
Chapter 1.8.2.2 --- Physical activity --- p.17
Chapter 1.8.2.3 --- Drug therapy --- p.18
Chapter CHAPTER 2 --- IMPAIRMENT OF OXIDIZED LDL ON ENDOTHELIUM-DEPENDENT RELAXATION
Chapter 2.1 --- Introduction --- p.19
Chapter 2.1.1 --- Properties and function of phenylephrine hydrochloride --- p.22
Chapter 2.1.2 --- Properties and function of acetylcholine --- p.22
Chapter 2.2 --- Objectives --- p.23
Chapter 2.3 --- Materials and methods --- p.24
Chapter 2.3.1 --- Preparation of drugs --- p.24
Chapter 2.3.2 --- Preparation of human native LDL --- p.25
Chapter 2.3.3 --- Preparation of oxidized LDL --- p.27
Chapter 2.3.4 --- Preparation of aorta --- p.27
Chapter 2.3.5 --- Measurement of Isometric Force in vitro --- p.30
Chapter 2.3.5.1 --- Protocol 1- Dose effect of oxidized LDL on acetylcholine-induced vasorelaxation --- p.30
Chapter 2.3.5.2 --- Protocol 2 - Time effect of oxidized LDL on acetylcholine-induced vasorelaxation --- p.30
Chapter 2.3.5.3 --- Protocol 3 - Effect of co-incubation of LDL and copper(ll) sulphate on acetylcholine-induced vasorelaxation --- p.31
Chapter 2.3.5.4 --- Protocol 4 - Effect of oxidized LDL on selected vasodilators --- p.32
Chapter 2.3.5.5 --- Protocol 5 - Effect of pretreatment of L-arginine on oxidized LDL impaired -endothelium-induced relaxation --- p.32
Chapter 2.3.5.6 --- Protocol 6 - Effect of a -tocopherol on oxidized LDL-damaged acetylcholine- induced vasorelaxation --- p.33
Chapter 2.3.5.7 --- Protocol 7 - Effect of a -tocopherol on LDL and copper(ll) sulphate- induced endothelial dysfunction --- p.33
Chapter 2.3.6 --- Western blot analysis of endothelial nitric oxide synthase (eNOS) protein --- p.34
Chapter 2.3.7 --- Statistics --- p.35
Chapter 2.4 --- Results --- p.36
Chapter 2.4.1 --- Dose effect of oxidized LDL on acetylcholine-induced vasorelaxation --- p.36
Chapter 2.4.2 --- Time effect of oxidized LDL on acetylcholine-induced vasorelaxation --- p.36
Chapter 2.4.3 --- Effect of co-incubation of LDL and copper(II) sulphate on acetylcholine- induced vasorelaxation --- p.39
Chapter 2.4.4 --- Effect of oxidized LDL on selected vasodilators --- p.41
Chapter 2.4.5 --- Effect of pretreatment of L-arginine on oxidized LDL impaired- acetylcholine-induced relaxation --- p.41
Chapter 2.4.6 --- Effect of a-tocopherol on oxidized LDL-damaged acetylcholine- induced vasorelaxation --- p.48
Chapter 2.4.7 --- Effect of a-tocopherol on LDL and copper(II) sulphate-induced endothelial dysfunction --- p.50
Chapter 2.4.8 --- eNOS Protein expression --- p.50
Chapter 2.5 --- Discussion --- p.53
Chapter CHAPTER 3 --- EFFECTS OF LDL INJECTION ON THE ENDOTHELIAL FUNCTION OF RATS
Chapter 3.1 --- Introduction --- p.58
Chapter 3.2 --- Objective --- p.60
Chapter 3.3 --- Methods and Materials --- p.61
Chapter 3.3.1 --- Preparation of Drugs --- p.61
Chapter 3.3.2 --- Preparation of LDL --- p.61
Chapter 3.3.3 --- Animal Treatment --- p.61
Chapter 3.3.4 --- Serum lipid and lipoprotein determinations --- p.62
Chapter 3.3.5 --- Measurement of serum MDA level by TBARS assay --- p.62
Chapter 3.3.6 --- Preparation of aorta --- p.62
Chapter 3.3.7 --- Organ bath experiment --- p.63
Chapter 3.3.8 --- Statistics --- p.64
Chapter 3.4 --- Result --- p.65
Chapter 3.4.1 --- Growth and food intake --- p.65
Chapter 3.4.2 --- "Effect of LDL injection on serum TC, TG and HDL-C" --- p.65
Chapter 3.4.3 --- Effect of LDL injection on non-HDL-C and ratio of non-HDL-C to HDL-C --- p.65
Chapter 3.4.4 --- Serum MDA level --- p.68
Chapter 3.4.5 --- Phenylephrine-induced contraction --- p.70
Chapter 3.4.6 --- Endothelium-dependent and -independent relaxation --- p.75
Chapter 3.5 --- Discussion --- p.79
Chapter CHAPTER 4 --- EFFECTS OF INDIVIDUAL COMPONENT OF OXIDIZED LDL ON ENDOTHELIUM-DEPENDENT RELAXATION
Chapter 4.1 --- Introduction --- p.83
Chapter 4.2 --- Objectives --- p.85
Chapter 4.3 --- Materials and methods --- p.86
Chapter 4.3.1 --- Preparation of drugs --- p.86
Chapter 4.3.2 --- Preparation of human native LDL and oxidized LDL --- p.86
Chapter 4.3.3 --- GC analysis of fatty acid composition in LDL --- p.86
Chapter 4.3.4 --- TBARS assay analysis of MDA content in LDL --- p.87
Chapter 4.3.5 --- GC analysis of cholesterol oxidation products in LDL --- p.89
Chapter 4.3.6 --- Thin-layer chromatography analysis of LPC in LDL --- p.91
Chapter 4.3.7 --- Preparation of aorta --- p.92
Chapter 4.3.8 --- Measurement of Isometric Force in vitro --- p.92
Chapter 4.3.8.1 --- Protocol 1- effect of LPC on acetylcholine-induced vasorelaxation --- p.92
Chapter 4.3.8.2 --- Protocol 2- effect of cholesterol oxidation products on acetylcholine-induced vasorelaxation --- p.92
Chapter 4.3.8.3 --- Protocol 3- effect of oxidized fatty acids on acetylcholine-induced vasorelaxation --- p.93
Chapter 4.3.9 --- Statistics --- p.93
Chapter 4.4 --- Results --- p.94
Chapter 4.4.1 --- Compositional differences between native LDL and oxidized LDL.… --- p.94
Chapter 4.4.2 --- Effect of LPC on endothelium-dependent relaxation --- p.98
Chapter 4.4.3 --- Effect of COPs on endothelium-dependent relaxation --- p.98
Chapter 4.4.4 --- Effect of oxidized fatty acids on endothelium-dependent relaxation --- p.101
Chapter 4.5 --- Discussion --- p.103
Chapter CHAPTER 5 --- EFFECTS OF DIETARY OXIDIZED CHOLESTEROL ON BLOOD CHOLESTEROL LEVEL IN HAMSTERS
Chapter 5.1 --- Introduction --- p.107
Chapter 5.2 --- Objectives --- p.111
Chapter 5.3 --- Materials and Methods --- p.112
Chapter 5.3.1 --- Preparation of Oxidized Cholesterol --- p.112
Chapter 5.3.2 --- Diet preparation --- p.112
Chapter 5.3.3 --- Animals --- p.113
Chapter 5.3.4 --- Serum lipid and lipoprotein determinations --- p.116
Chapter 5.3.5 --- GC analysis of cholesterol and cholesterol oxidation products on organs --- p.116
Chapter 5.3.6 --- Extraction of neutral and acidic sterols from fecal samples --- p.117
Chapter 5.3.6.1 --- Determination of neutral sterols --- p.117
Chapter 5.3.6.2 --- Determination of acidic sterols --- p.117
Chapter 5.3.6.3 --- GLC analysis of neutral and acidic sterols --- p.118
Chapter 5.3.7 --- Organ bath experiment --- p.121
Chapter 5.3.7.1 --- Preparation of aorta --- p.121
Chapter 5.3.7.2 --- Aortic relaxation --- p.121
Chapter 5.3.8 --- Analysis of the total area of atherosclerotic plaque on aorta --- p.122
Chapter 5.3.9 --- Statistics --- p.122
Chapter 5.4 --- Results --- p.123
Chapter 5.4.1 --- GC of oxidized cholesterol --- p.123
Chapter 5.4.2 --- Growth and food intake --- p.123
Chapter 5.4.3 --- "Effect of non-oxidized and oxidized cholesterol on serum TC, TG and HDL-C" --- p.123
Chapter 5.4.4 --- Effect of non-oxidized and oxidized cholesterol on non-HDL-C and ratio of non-HDL-C to HDL-C --- p.124
Chapter 5.4.5 --- Effect ofnon-oxidized and oxidized cholesterol on concentration of hepatic cholesterol --- p.128
Chapter 5.4.6 --- Effect of non-oxidized and oxidized cholesterol on concentration of cholesterol oxidation products accumulated in liver --- p.128
Chapter 5.4.7 --- Effect of non-oxidized and oxidized cholesterol on concentration of brain and aortic cholesterol --- p.128
Chapter 5.4.8 --- Effect of non-oxidized and oxidized cholesterol on fecal neutral and acidic sterols --- p.129
Chapter 5.4.9 --- Effect of non-oxidized and oxidized cholesterol on aortic relaxation --- p.135
Chapter 5.4.10 --- Effect of non-oxidzied and oxidized cholesterol on area of atherosclerotic plaque --- p.137
Chapter 5.5 --- Discussion --- p.139
Chapter CHAPTER 6 --- CONCLUSION --- p.143
REFERENCES --- p.146

Tuberculosis (TB) is the leading cause of death in individuals infected with human immunodeficiency virus (HIV) in several African countries, including South Africa. HIV-positive individuals do not have the immune system resources to keep TB in check and are as much as 30 times more likely to develop active TB than people who are HIV-negative. Many people infected with HIV develop TB as the first manifestation of AIDS and TB accelerates disease progression in HIV-positive individuals. HIV and TB pathogenesis are thus inextricably intertwined so that it is necessary for medical practitioners to have an understanding of the dynamics and treatment of HIV-TB coinfection. At present the question remains as to whether the best time for coinfected individuals to start antiretroviral treatment for HIV is at the beginning, the peak, or after the completion of the TB treatment phase. This dissertation was undertaken with the aim of obtaining some clarity on this question by creating a mathematical model of HIV-TB coinfection and its treatment. This needs an understanding of the biological interactions; therefore the dissertation begins with a discussion of the biological mechanisms of HIV, the human immune system, TB and the drug therapies for each disease. Thereafter a brief introduction to mathematical modelling reviews basic HIV models, which are then modified to include HIV drug therapy. Analyses and simulations of these models were carried out, which yielded some insights into the dynamics of HIV and HIV therapy. Finally HIV-TB coinfection is introduced by reviewing a previously developed model. Based on all the models reviewed, a model for coinfection is developed which includes treatment for HIV and TB. Numerical simulations suggest that, if HIV disease progression is at an advanced stage of the immune system collapsing towards AIDS, with low T-cell count and high viral load, it is necessary to treat for both diseases simultaneously to ensure a positive survival prognosis for the coinfected individual. However, if disease progression is in the early stages of AIDS, with T-cell count and viral load beginning to display signs of the immune system collapse but still at reasonable levels relative to advanced stages, it need not be necessary to treat both diseases simultaneously. TB can be treated first, and upon completion HIV treatment can be initiated thus sparing the coinfected individual from the compounded side-effects and drug-drug interactions which usually result from simultaneous treatment.
Thesis (M.Sc.)-University of KwaZulu-Natal, 2007.

by Choi, Heung Ling.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1994.
Includes bibliographical references (leaves 79-95).
Acknowledgement --- p.iv
Abstract --- p.vi
Chapter CHAPTER 1 --- INTRODUCTION
Chapter 1.1 --- Cocaine --- p.1
Chapter 1.1.1 --- History --- p.1
Chapter 1.1.2 --- Epidemiology --- p.2
Chapter 1.1.3 --- Pharmacology --- p.3
Chapter 1.2 --- Maternal Cocaine Abuse --- p.5
Chapter 1.2.1 --- Human Studies --- p.5
Chapter 1.2.1.1 --- Prevalence --- p.5
Chapter 1.2.1.2 --- Effects of Cocaine on the Developing Fetus --- p.7
Chapter 1.2.1.2.1 --- Fetal Mortality --- p.8
Chapter 1.2.1.2.2 --- Placental Abruption --- p.9
Chapter 1.2.1.2.3 --- Premature Birth --- p.9
Chapter 1.2.1.2.4 --- Neonatal Effects --- p.10
Chapter 1.2.1.3 --- Congenital Abnormalities --- p.11
Chapter 1.2.1.3.1 --- Cardiovascular Abnormality --- p.11
Chapter 1.2.1.3.2 --- Genitourinary Tract Malformation --- p.12
Chapter 1.2.1.3.3 --- Gastrointestinal Abnormality --- p.12
Chapter 1.2.1.3.4 --- Respiratory Disorders --- p.13
Chapter 1.2.1.3.5 --- Visual and Hearing Disorders --- p.14
Chapter 1.2.1.3.6 --- CNS and Behavioural Abnormalities --- p.15
Chapter 1.2.2 --- Animal Studies --- p.17
Chapter 1.2.2.1 --- "Routes of Administration, Dosage and Tissue Distribution " --- p.18
Chapter 1.2.2.2 --- Maternal and Offspring Effects --- p.21
Chapter 1.2.2.2.1 --- Fetal and Maternal Mortality --- p.22
Chapter 1.2.2.2.2 --- Gestational Length --- p.22
Chapter 1.2.2.2.3 --- Maternal Weight Gain and Fetal Weight --- p.23
Chapter 1.2.2.2.4 --- Little Size --- p.24
Chapter 1.2.2.3 --- Congenital Abnormalities --- p.24
Chapter 1.2.2.4 --- Behavioral Changes --- p.26
Chapter 1.2.2.5 --- Neurochemical Changes --- p.28
Chapter 1.2.2.5.1 --- Glucose Metabolism --- p.28
Chapter 1.2.2.5.2 --- Dopamine Transporter --- p.29
Chapter 1.2.2.5.3 --- Dopamine D1 Receptor --- p.29
Chapter 1.2.2.5.4 --- Dopamine D2 Receptor --- p.30
Chapter 1.2.2.5.5 --- Tyrosine Hydroxylase --- p.30
Chapter 1.2.2.5.6 --- Other Changes --- p.31
Chapter 1.3 --- The Aim of the Study --- p.31
Chapter CHAPTER II --- MATERIALS AND METHODS
Chapter 2.1 --- Administration of Cocaine --- p.34
Chapter 2.2 --- Biochemical Studies --- p.35
Chapter 2.2.1 --- Receptor Binding Assays --- p.36
Chapter 2.2.1.1 --- Dopamine Transporter --- p.37
Chapter 2.2.1.1.1 --- Specific Binding Assay and Scatchard Analysis --- p.37
Chapter 2.2.1.2 --- Dopamine D1 Receptor --- p.38
Chapter 2.2.1.2.1 --- Association Curve --- p.38
Chapter 2.2.1.2.2 --- Competition Assay --- p.39
Chapter 2.2.1.2.3 --- Specific Binding Assay and Scatchard Analysis --- p.39
Chapter 2.2.1.3 --- Dopamine D2 Receptor --- p.39
Chapter 2.2.1.3.1 --- Association Curve --- p.40
Chapter 2.2.1.3.2 --- Competition Assay --- p.40
Chapter 2.2.1.3.3 --- Specific Binding Assay and Scatchard Analysis --- p.40
Chapter 2.2.1.4 --- Assay for Residual Cocaine in Maternal Brain --- p.41
Chapter 2.3 --- Statistics --- p.42
Chapter 2.4 --- Morphological Studies --- p.42
Chapter 2.4.1 --- Tyrosine Hydroxylase (TH) Immunocytochemical Staining --- p.42
Chapter 2.5 --- Molecular Genetic Studies --- p.44
Chapter 2.5.1 --- Material for DNA Insert --- p.44
Chapter 2.5.1.1 --- "Dopamine Transporter, D2 receptor and β-actin cDNA Probe " --- p.44
Chapter 2.5.2 --- Preparation for DNA Insert --- p.45
Chapter 2.5.2.1 --- Competent Cells and Transformation of Plasmid --- p.45
Chapter 2.5.2.2 --- Growth Transformed Bacteria and Isolation of DNA --- p.46
Chapter 2.5.2.3 --- Purification of cDNA by Geneclean® II Kit --- p.47
Chapter 2.5.3 --- Isolation of Total mRNA From Tissue --- p.47
Chapter 2.5.4 --- Northern Blot Analysis --- p.48
Chapter 2.5.4.1 --- Analysis of Northern Blots --- p.50
Chapter 2.5.5 --- In Situ Hybridization --- p.50
Chapter 2.5.5.1 --- Tissue Preparation --- p.50
Chapter 2.5.5.2 --- Preparation of Dopamine Transporter Ribroprobe …… --- p.50
Chapter 2.5.5.3 --- In Situ Hybridization Histochemistry --- p.51
Chapter CHAPTER III --- RESULTS
Chapter 3.1 --- "Litter Size, Birth Weight and Maternal Weight Gain " --- p.53
Chapter 3.2 --- Biochemical Studies --- p.53
Chapter 3.2.1 --- Specific Binding --- p.53
Chapter 3.2.2 --- Dopamine Transporter - Scatchard Analysis --- p.54
Chapter 3.2.3 --- Dopamine Receptor --- p.55
Chapter 3.2.3.1 --- Association Curve --- p.56
Chapter 3.2.3.2 --- Competitive Curve --- p.57
Chapter 3.2.3.3 --- Scatchard Analysis --- p.57
Chapter 3.2.4 --- Dopamine D2 Receptor --- p.59
Chapter 3.2.4.1 --- Association Curve --- p.59
Chapter 3.2.4.2 --- Competitive Curve --- p.59
Chapter 3.2.4.3 --- Scatchard Analysis --- p.59
Chapter 3.2.5 --- Residual Cocaine Assay in Maternal Brain --- p.61
Chapter 3.2.5.1 --- Specific Binding --- p.61
Chapter 3.2.5.1.1 --- Dopamine Transporter --- p.61
Chapter 3.3.5.1.2 --- Dopamine D1 Receptor --- p.62
Chapter 3.3.5.1.3 --- Dopamine D2 Receptor --- p.62
Chapter 3.3 --- Morphological Studies --- p.62
Chapter 3.3.1 --- Tyrosine Hydroxylase (TH) Immunocytochemical Staining --- p.62
Chapter 3.4 --- Molecular Genetic Studies --- p.63
Chapter 3.4.1 --- Northern Blot Analysis --- p.63
Chapter 3.4.1.1 --- Dopamine Transporter --- p.63
Chapter 3.4.1.2 --- Dopamine D2 Receptor --- p.64
Chapter 3.4.2 --- In Situ Hybridization --- p.64
Chapter CHAPTER IV --- DISCUSSION AND CONCLUSION
Chapter 4.1 --- Discussion --- p.65
Chapter 4.2 --- Conclusion --- p.77
References --- p.79
Publications --- p.95

丹參和葛根為我國民間常用的傳統藥材, 常用於心血管疾病的治療。本試驗主要研究丹參葛根複方(DG, 7:3)對大鼠基底動脈的舒張作用 及腦保護作用。
上述所有藥物對U46619預收縮的基底動脈環呈現濃度依賴性的舒張作用。一氧化氮合酶抑制劑L-NAME以及鳥苷酸環化酶抑制劑ODG部分抑制葛根素的舒張作用。在另一組去內皮試驗中, 腺苷酸環化酶抑制劑SQ22536, 鳥苷酸環化酶抑制劑ODG, 大電導鈣離子依賴型鉀通道抑制劑Iberiotoxin以及電壓門控型鉀通道抑制劑4-AP對所有藥物的舒張作用沒有影響。然而， ATP型鉀通道抑制劑格列本脲能夠抑制丹參葛根複方，丹參，葛根，丹參素，葛根素，大豆苷以及大豆苷元的最大舒張反應。內向整流型鉀通道抑制劑氯化鋇則降低丹參酚酸B和大豆苷元的最大反應值。非選擇性鉀通道抑制劑乙基氯化銨以及所有鉀通道抑制劑的混合物顯著抑制上述所有藥物的舒張作用。除了葛根素之外，所有的藥物動度依賴性的抑制氯化鈣所引起的血管收縮。
體內研究發現大鼠經歷10分鐘雙側頸總動脈夾閉合並低血壓，及24小時的複灌後，與假手術組動物相比，腦血流量顯著降低，氧化性損傷明顯可見。連續7天口服丹參葛根複方（0.3g/kg 和 3g/kg）, 丹參 (3g/kg)，或者葛根 (3g/kg)對血壓沒有影響。但是，高劑量的丹參葛根複方 (3g/kg) 能夠提高超氧化物歧化酶和過氧化氫酶的活性，抑制丙二醛和一氧化氮的產生。3g/kg的葛根可以提高超氧化物歧化酶的活性，3g/kg的丹參則能抑制一氧化氮的產生。在大鼠中動脈阻塞模型中，連續7天口服丹參葛根複方（3g/kg）能明顯降低腦部的梗死率，同時改善大鼠的神經行為學。
總體來說，研究發現丹參葛根複方，丹參，葛根，丹參酚酸B，大豆苷以及大豆苷元的血管舒張作用是通過開平滑肌細胞的通鉀離子通道以及抑制鈣離子內流而實現的。然而葛根素的血管舒張作用是內皮依賴性的，通過產生一氧化氮，開放平滑肌細胞的鉀離子通道而實現的。丹參葛根複方能起到一定的腦保護作用。總而言之，研究表明上述藥物可能會對阻塞性腦血管病的人群有益處。
Danshen and gegen are used in traditional Chinese medicine for the treatment of cardiovascular diseases. In this study, the relaxant actions of a danshen and gegen formulation (DG; ratio 7:3) and its constituents were investigated on rat-isolated cerebral basilar artery. In addition, the neuroprotective effect of DG was explored in rats subjected to global and focal ischaemia.
DG and all its constituents produced concentration-dependent relaxation of the artery rings precontracted by U46619. Removal of the endothelium had no effect on their vasodilator actions except the maximum response (I[subscript max]) to puerarin was inhibited by 42%. The nitric oxide synthase (NOS) inhibitor L-NAME and guanylyl cyclase (GC) inhibitor ODQ but not the cyclo-oxygenase (COX) inhibitor flurbiprofen produced partial inhibition on the puerarin-induced effect. In a set of endothelium-denuded artery rings, adenylyl cyclase (AC) inhibitor SQ22536, GC inhibitor ODQ, KV channel inhibitor 4-aminopyridine (4-AP) and BK[subscript Ca) channel inhibitor iberiotoxin had no influence on their vasodilator actions. However, pretreatment with K[subscript ATP] channel inhibitor glibenclamide reduced Imax to DG, danshen, gegen, danshensu, puerarin, daidzein and daidzin. K[subscript IR] inhibitor barium chloride (BaCl₂) reduced II[subscript max] to salvianolic acid B and daidzein. The non-selective K⁺ channel inhibitor tetraethylammonium (TEA), or a combination of all the K⁺ channel inhibitors produced significant partial inhibitions on all the agents’ actions. Electrophysiological studies on smooth muscle cells isolated from rat basilar artery also confirmed that DG, danshen, gegen danshensu, puerarin, daidzein and daidzin elevated K[subscript ATP] currents. In addition, DG and all its constituents, except puerarin, produced concentration-dependent inhibition on CaCl₂-induced vasoconstrictions. These findings were confirmed by con-focal microscopy studies.
In vivo study on a rat model of global ischaemia showed that challenging the rats with 10 min bilateral common carotid artery occlusion combined with hypotension, and followed by 24 h reperfusion produced significant decrease in cerebral blood flow and oxidative damage compared to sham-operated animals. Administration of DG (0.3 g/kg and 3 g/kg, p.o.), danshen (3 g/kg, p.o.) or gegen (3 g/kg, p.o.) for 7 days had no effect on blood pressure. However, the 7 days treatment with DG (3 g/kg) restored superoxide dismutase (SOD) and catalase (CAT) activities, suppressed the production of maleic dialdehyde (MDA), and inhibited the production of nitric oxide (NO). In addition, gegen (3 g/kg) restored SOD enzyme activity, whereas, danshen (3 g/kg) inhibited NO production. In addition, treatment with DG (3 g/kg) showed a significant reduction in infarct weight and improved the neurological deficit in a rat model of focal cerebral ischaemia induced by middle cerebral artery occlusion (MCAO).
In conclusion, the vasorelaxant actions of DG, danshen, gegen, salvianolic acid B, danshensu, daidzein and daidzin were found to involve the opening of K⁺ channels and inhibition of Ca²⁺ influx in the vascular smooth muscle cells. In contrast, puerarin produced vasodilatation via an endothelium-dependent mechanism involving NO production and an endothelium-independent pathway mediated by the opening of K⁺ channels. DG may have some cerebro-protective effects. Overall, the present studies showed that DG and its constituents could be beneficial to patients with obstructive cerebrovascular diseases.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Deng, Yan.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2012.
Includes bibliographical references (leaves 164-178).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.
ABSTRACT --- p.v
摘要 --- p.iviii
ACKNOWLEDGEMENTS --- p.x
PUBLICATIONS BASED ON THIS THESIS --- p.xii
ABBREVIATIONS --- p.xiv
Chapter CHAPTER 1 --- Introduction --- p.1
Chapter 1.1 --- Chinese Medicines in treatment of cerebrovascular diseases --- p.2
Chapter 1.2 --- Danshen --- p.4
Chapter 1.2.1 --- Chemical constituents --- p.4
Chapter 1.2.1.1 --- Hydrophilic compounds of danshen --- p.4
Chapter 1.2.1.2 --- Lipophilic compounds of danshen --- p.5
Chapter 1.2.1.3 --- Other compounds --- p.5
Chapter 1.2.2 --- Pharmacological activities --- p.5
Chapter 1.2.2.1 --- Vascular protection --- p.5
Chapter 1.2.2.2 --- Anti-tumour --- p.7
Chapter 1.2.2.3 --- Treatment of liver diseases --- p.8
Chapter 1.2.2.4 --- Treatment of drug addiction --- p.9
Chapter 1.2.2.5 --- Treatment of kidney diseases --- p.10
Chapter 1.2.3 --- Pharmacokinetics --- p.10
Chapter 1.3 --- Gegen --- p.12
Chapter 1.3.1 --- Chemical constituents --- p.12
Chapter 1.3.2 --- Pharmacology --- p.13
Chapter 1.3.2.1 --- Vascular effects --- p.13
Chapter 1.3.2.2 --- Anti-diabetes --- p.14
Chapter 1.3.2.3 --- Anti-hypercholesterolaemia --- p.15
Chapter 1.3.2.4 --- Anti-inflammation --- p.16
Chapter 1.3.2.5 --- Anti-platelet aggregation --- p.17
Chapter 1.3.3 --- Pharmacokinetics --- p.17
Chapter 1.4 --- Danshen and gegen formulation --- p.19
Chapter 1.5 --- Mechanisms of vasodilatation --- p.22
Chapter 1.5.1 --- Endothelium derived relaxant factors (EDRFs) --- p.22
Chapter 1.5.1.1 --- Nitric oxide (NO) --- p.22
Chapter 1.5.1.2 --- Prostacyclin (PGI₂) --- p.23
Chapter 1.5.1.3 --- Endothelium-derived hyperpolarizating factors (EDHFs)- --- p.23
Chapter 1.5.2 --- Signal transduction pathways --- p.24
Chapter 1.5.2.1 --- Guanylyl cyclase-cGMP pathway --- p.24
Chapter 1.5.2.2 --- Adenylyl cyclase-cAMP pathway --- p.24
Chapter 1.5.3 --- Ion channels --- p.25
Chapter 1.5.3.1 --- Potassium channels (K⁺ channels) --- p.25
Chapter 1.5.3.2 --- Calcium channel (Ca²⁺ channels) --- p.25
Chapter 1.6 --- Aims of study --- p.27
Chapter CHAPTER 2 --- Materials and method --- p.28
Chapter 2.1 --- Herbal preparation --- p.28
Chapter 2.1.1 --- DG, danshen and gegen preparation --- p.28
Chapter 2.1.2 --- Identification and quantification of chemical markers in DG water extract --- p.29
Chapter 2.2 --- Experiments on rat basilar artery --- p.30
Chapter 2.2.1 --- Animals --- p.30
Chapter 2.2.2 --- Chemicals --- p.30
Chapter 2.2.3 --- Isolation and mounting of blood vessels --- p.33
Chapter 2.2.4 --- Protocols --- p.34
Chapter 2.2.4.1 --- Effects on U46619-precontracted tone --- p.34
Chapter 2.2.4.2 --- Endothelium-dependent mechanism --- p.34
Chapter 2.2.4.3 --- Endothelium-independent mechanism --- p.35
Chapter 2.2.4.4 --- Calcium channels --- p.36
Chapter 2.2.4.5 --- Positive control --- p.36
Chapter 2.2.5 --- Statistical analysis --- p.37
Chapter 2.3 --- Experiments on rat cerebral basilar artery smooth muscle cells K[subscript ATP] channals --- p.38
Chapter 2.3.1 --- Animals --- p.38
Chapter 2.3.2 --- Chemicals --- p.38
Chapter 2.3.3 --- Isolation of rat cerebral vascular smooth muscle cells --- p.40
Chapter 2.3.4 --- Whole cell patch-clamp electrophysiology --- p.40
Chapter 2.3.5 --- Statistical analysis --- p.44
Chapter 2.4 --- Experiments on rat cerebral basilar artery smooth muscle cells calcium channels --- p.45
Chapter 2.4.1 --- Animals --- p.45
Chapter 2.4.2 --- Chemicals --- p.45
Chapter 2.4.3 --- Isolation of rat cerebral vascular smooth muscle cells --- p.47
Chapter 2.4.4 --- Dye loading and determination of [Ca²⁺]i --- p.47
Chapter 2.4.5 --- Statistical analysis --- p.48
Chapter 2.5 --- In vivo study of global ischaemia --- p.49
Chapter 2.5.1 --- Animals --- p.49
Chapter 2.5.2 --- Drugs and chemicals --- p.49
Chapter 2.5.3 --- Experimental protocols for global ischaemia --- p.49
Chapter 2.5.4 --- Induction of global ischaemia --- p.50
Chapter 2.5.5 --- Blood pressure measurement --- p.52
Chapter 2.5.6 --- Measurement of cerebral blood flow --- p.52
Chapter 2.5.7 --- Biochemical assessment --- p.53
Chapter 2.5.7.1. --- Dissection and homogenization --- p.53
Chapter 2.5.7.2. --- Measurement of malondialdehyde (MDA) --- p.53
Chapter 2.5.7.3. --- Estimation of nitrite --- p.53
Chapter 2.5.7.4 --- Superoxide dismutase activity (SOD) --- p.54
Chapter 2.5.7.5 --- Reduced glutathione (GSH) --- p.54
Chapter 2.5.7.6 --- Catalase (CAT) --- p.55
Chapter 2.5.7.7 --- NOS activity --- p.55
Chapter 2.5.7.8 --- Protein --- p.56
Chapter 2.5.8 --- Statistical analysis --- p.56
Chapter 2.6 --- In vivo study of focal ischaemia --- p.57
Chapter 2.6.1 --- Animals --- p.57
Chapter 2.6.2 --- Drugs and chemicals --- p.57
Chapter 2.6.3 --- Experimental protocols for global ischaemia --- p.57
Chapter 2.6.4 --- Focal cerebral ischaemia-reperfusion model --- p.57
Chapter 2.6.5 --- Assessment of neurobehavioural changes --- p.59
Chapter 2.6.6 --- Assessment of cerebral infarction --- p.60
Chapter 2.6.7 --- Statistical analysis --- p.60
Chapter CHAPTER 3 --- Results --- p.61
Chapter 3.1 --- Identification and quantification of chemical markers in DG water extract --- p.61
Chapter 3.2 --- Effects of DG and its constituents on rat cerebral basilar artery --- p.64
Chapter 3.2.1 --- Investigations on endothelium-dependent mechanisms --- p.64
Chapter 3.2.2 --- Investigations on endothelium-independent mechanisms --- p.68
Chapter 3.2.3 --- Positive control --- p.86
Chapter 3.2.3 --- Investigations on calcium channels --- p.88
Chapter 3.3 --- Effects of DG and its constituents on rat cerebral basilar artery smooth muscle cells --- p.91
Chapter 3.3.1 --- Effects of water crude-extracts of DG, danshen, and gegen on K[subscript ATP] channels --- p.91
Chapter 3.3.2 --- Effects of active constituents of danshen hydrophilic fraction on K[subscript ATP] channels --- p.100
Chapter 3.3.3 --- Effects of the major isoflavonoids of gegen on K[subscript ATP] channels --- p.105
Chapter 3.4 --- Effects of DG and its constituents on calcium channels of basilar artery smooth muscle cells --- p.112
Chapter 3.5 --- Effects of DG, danshen and gegen on rat global ischaemia --- p.117
Chapter 3.5.1 --- Effects of DG, danshen and gegen on rats’ blood pressure and cerebral blood flow --- p.117
Chapter 3.5.2 --- Effects of DG, danshen and gegen on lipid peroxidation --- p.120
Chapter 3.5.3 --- Effects of DG, danshen and gegen on SOD activity --- p.120
Chapter 3.5.4 --- Effects of DG, danshen and gegen on CAT activity --- p.120
Chapter 3.5.5 --- Effects of DG, danshen and gegen on reduced GSH level --- p.121
Chapter 3.5.6 --- Effects of DG, danshen and gegen on NOS system --- p.126
Chapter 3.6 --- Effect of DG on rat focal ischaemia --- p.129
Chapter 3.6.1 --- Effect of DG on cerebral infarction --- p.129
Chapter 3.6.2 --- Effect of DG on neurological deficits --- p.129
Chapter CHAPTER 4 --- Discussion --- p.132
Chapter 4.1 --- Studies of DG and its constituents on rat cerebral basilar artery --- p.133
Chapter 4.1.1 --- Constituents of DG on U46619-precontracted tone --- p.133
Chapter 4.1.2 --- Investigations on endothelium-dependent mechanisms --- p.133
Chapter 4.1.3 --- Investigations on endothelium-independent mechanisms --- p.136
Chapter 4.1.4 --- Investigations on calcium channels --- p.139
Chapter 4.2 --- Effects of DG and its constituents on rat cerebral basilar artery smooth muscle cell K[subscript ATP] channels --- p.143
Chapter 4.3 --- Effects of DG and its constituents on calcium influx in rat basilar artery smooth muscle cells --- p.147
Chapter 4.4 --- Effects of DG, danshen and gegen on rat transient global ischaemia --- p.150
Chapter 4.4.1 --- Effects of DG, danshen and gegen on rats’ blood pressure and cerebral blood flow --- p.150
Chapter 4.4.2 --- Effects of DG, danshen and gegen on lipid peroxidation, SOD and CAT activity, and GSH level --- p.152
Chapter 4.4.3 --- Effects of DG, danshen and gegen on NOS system --- p.155
Chapter 4.5 --- Effects of DG on rat focal ischaemia --- p.157
Chapter 4.6 --- Further studies --- p.160
Chapter 4.7 --- Conclusion --- p.162
REFERENCES --- p.164

碩士
南華大學
非營利事業管理研究所
94
Illegal drug-using of abuse arouses great concerns for the public as always, and also utmost improvement for Taiwan Police Agency. According to the record from Taiwan After-care Association, the recidivism percentage of drug-using reaches highly 67.6%, that is the reason to research the effect on drug on drug-using offenders. 　 　　In this study, the drug of offenders consulted by Taiwan After-care Association, Taichung Branch as models, to make a questionnaire for the scope of research. Total 166 questionnaires we distribute 166 valid questionnaires and 4 invalid questionnaires, applied for the reliable analysis, descriptive statistics, one-way ANOVE, Duncan”s test and T test etc, to conduct the description and assumption of results. 　 　　The obvious discrepancy between ”marriage” and occupation before abstention” is revealed in the personal background of Importance and Satisfaction; as well as using drug, ”abstention times”, “drug-using motivation”, drug-using frequency”,” drug-using duration” and “facing problems” produce the obvious differences. 　 　　The effect on After-care support and counseling is different from “occupation before abstention” in the personal background, as well as using drug, ”abstention times”, “drug-using motivation”, drug-using frequency” and “drug-using duration” produce the obvious differences. 　 　　According to the results of this study, the recommendations are: 1.the process of counseling clients: For the purpose of improving the effects on drug offenders, relative assistance and diagnosis are necessary; 2.the recruitment and training of drug quitting counselor: The increasing of psychologist and social worker who execute the psychological treatment and therapy allocated in drug quitting center or After-care Association will effect the success of drug quitting; 3.Anti-durg advertisement: To stress on the defect and law problem and to appeal for rejecting drug-using are subject to reduce the curiosity of drug for the public; 4. The drug abusers of occupation and requested for arranging occupation: To strengthen occupational counseling acquire a stable vocation; 5.The long period and high frequency of using drug abusers: Should be execute special psychological counseling and emphasized on law-related education; 6.Continuous tracing and counseling: To support drug-using offenders away from drug is aimed to reinforce the determination of rejecting drug.

Leung Lai-Kin.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2003.
Includes bibliographical references (leaves 110-113).
Abstracts in English and Chinese.
Abstract English --- p.i
Chinese --- p.iii
Acknowledgments --- p.v
Table of contents --- p.vi
List of Tables --- p.ix
List of Figures --- p.x
List of Abbreviations --- p.xiii
Chapter Chapter 1 --- Introduction --- p.1
Chapter Chapter 2 --- Establishment of compound formulation --- p.4
Chapter 2.1 --- Formulation research --- p.4
Chapter 2.2 --- Aqueous extract preparation --- p.6
Chapter 2.2.1 --- Materials and Methods --- p.6
Chapter 2.2.2 --- Results --- p.7
Chapter 2.2.3 --- Discussion --- p.9
Chapter 2.3 --- Preliminary test --- p.10
Chapter 2.3.1 --- Materials and Methods --- p.10
Chapter 2.3.2 --- Results --- p.12
Chapter 2.3.3 --- Discussion --- p.14
Chapter Chapter 3 --- Quality Control --- p.15
Chapter 3.1 --- HPLC standardization --- p.16
Chapter 3.2 --- Materials and Methods --- p.19
Chapter 3.3 --- Results --- p.20
Chapter 3.4 --- Discussion --- p.28
Chapter Chapter 4 --- Antioxidant study --- p.29
Chapter 4.1 --- Red blood cell hemolysis model --- p.30
Chapter 4.1.1 --- Materials and Methods --- p.30
Chapter 4.1.2 --- Results --- p.30
Chapter 4.1.3 --- Discussion --- p.32
Chapter 4.2 --- Ischemia-Reperfusion on Langendorff --- p.33
Chapter 4.2.1 --- Materials and Methods --- p.34
Chapter 4.2.2 --- Results --- p.37
Chapter 4.2.3 --- Discussion --- p.60
Chapter Chapter 5 --- Vasodilation study --- p.61
Chapter 5.1 --- Vasodilation in organ bath --- p.63
Chapter 5.1.1 --- Materials and Methods --- p.63
Chapter 5.1.2 --- Results --- p.65
Chapter 5.1.3 --- Discussion --- p.79
Chapter 5.2 --- Endothelium dependent vasodilation --- p.80
Chapter 5.2.1 --- Materials and Methods --- p.80
Chapter 5.2.2 --- Results --- p.83
Chapter 5.2.3 --- Discussion --- p.95
Chapter Chapter 6 --- Anti-platelet study --- p.96
Chapter 6.1 --- CFU-MK plasma clot colony assay --- p.97
Chapter 6.2 --- Materials and Methods --- p.97
Chapter 6.3 --- Results --- p.100
Chapter 6.4 --- Discussion --- p.103
Chapter Chapter 7 --- Discussions and prospects --- p.104
Chapter 7.1 --- Discussions --- p.104
Chapter 7.2 --- Prospects --- p.107
Chapter 7.2.1 --- TCM capsule with GMP --- p.107
Chapter 7.2.2 --- Clinical Trial of the capsule --- p.109
References --- p.110

Tse Pui Shan.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2003.
Includes bibliographical references (leaves [191]-216).
Abstracts in English and Chinese.
ACKNOWLEDGEMENTS --- p.I
ABBREVIATIONS --- p.III
ABSTRACT --- p.VIII
摘要 --- p.X
PUBLICATIONS --- p.XII
TABLE OF CONTENTS --- p.XIII
Chapter CHAPTER 1: --- GENERAL INTRODUCTION
Chapter 1.1 --- Human Immune System and Cancer --- p.1
Chapter 1.1.1 --- Brief Introduction of the Human Immune System --- p.1
Chapter 1.1.2 --- Prevalence of Cancer in Hong Kong --- p.4
Chapter 1.1.3 --- The Role of the Immune System in Tumorigenesis --- p.4
Chapter 1.1.4 --- Cancer Treatment --- p.5
Chapter 1.1.5 --- Cancer Prevention --- p.5
Chapter 1.2 --- Mushroom Polysaccharides --- p.6
Chapter 1.2.1 --- General Aspects of Mushroom Polysaccharides --- p.6
Chapter 1.2.2 --- Structure of Mushroom Polysaccharides --- p.9
Chapter 1.2.2.1 --- Beta (P)-D-glucans --- p.9
Chapter 1.2.2.2 --- Heteroglucans and Protein-bound Polysaccharides --- p.10
Chapter 1.2.2.3 --- Structure-Function Interactions of Polysaccharides --- p.12
Chapter 1.2.3 --- Molecular Interactions of Polysaccharides --- p.14
Chapter 1.2.4 --- Biological Activities of Polysaccharides --- p.15
Chapter 1.2.4.1 --- Anti-tumor Activities of Polysaccharides --- p.15
Chapter 1.2.4.2 --- Immunomodulatory Activities of Polysaccharides --- p.16
Chapter 1.3 --- Yun Zhi (Coriolus versicolor) --- p.17
Chapter 1.3.1 --- General Features of Yun Zhi --- p.17
Chapter 1.3.2 --- Traditional Uses of Yun Zhi --- p.20
Chapter 1.3.3 --- Active Ingredients of Yun Zhi --- p.20
Chapter 1.3.3.1 --- "Origin, Properties and Composition of PSK" --- p.21
Chapter 1.3.3.2 --- "Origin, Properties and Composition of PSP" --- p.22
Chapter 1.3.4 --- Pharmacological Actions of PSP and PSK --- p.25
Chapter 1.3.4.1 --- Immunomodulatory Activities --- p.25
Chapter 1.3.4.2 --- Anti-tumor Activities --- p.32
Chapter 1.3.4.2 --- Antiviral and Antimicrobial Activities --- p.35
Chapter 1.3.4.3 --- Antioxidant Activities --- p.36
Chapter 1.3.5 --- Human Clinical Studies on Yun Zhi --- p.36
Chapter 1.3.6 --- Toxicology of Yun Zhi --- p.42
Chapter 1.4 --- Danshen (Salvia miltiorrhiza) --- p.43
Chapter 1.4.1 --- General Features of Danshen --- p.43
Chapter 1.4.2 --- Traditional Uses of Danshen --- p.46
Chapter 1.4.3 --- Active Ingredients of Danshen --- p.47
Chapter 1.4.4 --- Pharmacological Actions of Danshen --- p.50
Chapter 1.4.4.1 --- Cardiovascular Effects --- p.50
Chapter 1.4.4.2 --- Scavenging Effects on Free Radicals --- p.52
Chapter 1.4.4.3 --- Hepatoprotective Effects --- p.54
Chapter 1.4.4.4 --- Anti-tumor Effects --- p.56
Chapter 1.4.4.5 --- Renal Protective Effects --- p.56
Chapter 1.4.5 --- Human Clinical Studies --- p.57
Chapter 1.4.6 --- Toxicity of Danshen --- p.59
Chapter 1.5 --- Aims and Scopes of This Investigation --- p.60
Chapter CHAPTER 2: --- MATERIALS AND METHODS
Chapter 2.1 --- Normal Subjects --- p.62
Chapter 2.1.1 --- Inclusion and Exclusion Criteria of Recruitment --- p.62
Chapter 2.1.2 --- Study Design and Procedure --- p.63
Chapter 2.1.3 --- Treatment and Blinding --- p.65
Chapter 2.1.4 --- Blood Sampling --- p.66
Chapter 2.1.5 --- Blood Processing for Assessment of Immunological Functions --- p.67
Chapter 2.2 --- Materials --- p.69
Chapter 2.2.1 --- Endotoxin Assay --- p.69
Chapter 2.2.2 --- Reagents for Whole Blood Assay --- p.69
Chapter 2.2.2.1 --- Plain RPMI 1640 Medium --- p.69
Chapter 2.2.2.2 --- Phosphate-Buffered Saline (PBS) --- p.69
Chapter 2.2.2.3 --- Mitogens --- p.70
Chapter 2.2.3 --- Reagents for Total RNA Extraction --- p.70
Chapter 2.2.3.1 --- Ficoll-Paque Density Gradient Solution --- p.70
Chapter 2.2.3.2 --- RNA Extraction Kit --- p.70
Chapter 2.2.3.3 --- RNase-Free DNase Set --- p.71
Chapter 2.2.3.4 --- β-Mercaptoethanol (β-ME) Solution --- p.71
Chapter 2.2.4 --- Reagents for Flow Cytometric Analysis of T/B/NK Cell Ratios --- p.71
Chapter 2.2.4.1 --- MultiTEST IMK Kit with TruCOUNT Tubes --- p.71
Chapter 2.2.4.2 --- FACSFlo´wёØ Sheath Fluid --- p.74
Chapter 2.2.4.3 --- CaliBRITE 3 and APC Beads --- p.74
Chapter 2.2.5 --- Immunoassay Kits for Measuring Cytokines Level --- p.75
Chapter 2.2.5.1 --- Enzyme-linked Immunosorbent Assay (ELISA) Kits of Cytokines --- p.75
Chapter 2.2.5.2 --- Human Thl/Th2 Cytokine Cytometric Bead Array (CBA) Kit-II --- p.75
Chapter 2.2.6 --- Reagents and Buffers for Gel Electrophoresis --- p.78
Chapter 2.2.6.1 --- Ethidium Bromide (EtBr) --- p.78
Chapter 2.2.6.2 --- Gel Loading Solution (5X) --- p.78
Chapter 2.2.6.3 --- Tris-Acetate-EDTA (TAE) Buffer --- p.78
Chapter 2.2.6.4 --- Agarose Gel --- p.78
Chapter 2.2.6.5 --- 100 base pair DNA Ladder --- p.79
Chapter 2.2.7 --- Kits and Reagents for Messenger RNA (mRNA) Expression Array --- p.79
Chapter 2.2.7.1 --- Human Inflammatory Cytokine/Receptor GEArraýёØ Q Series Kit --- p.79
Chapter 2.2.7.2 --- Deoxynucleoside Triphosphates (dNTPs) --- p.84
Chapter 2.2.7.3 --- Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLVRT) --- p.84
Chapter 2.2.7.4 --- Rnasin Ribonuclease Inhibitor --- p.84
Chapter 2.2.7.5 --- Biotin-16-2'-deoxy-uridine-5'-triphosphate (Biotin-16-dUTP) --- p.85
Chapter 2.2.7.6 --- Salmon Sperm DNA Solution --- p.85
Chapter 2.2.7.7 --- 100 % Sodium Dodecyl Sulfate (SDS) Solution --- p.86
Chapter 2.2.7.8 --- 20X SSC --- p.86
Chapter 2.2.7.9 --- ECL Films (Hyperfilm 226}0ёØ ECL 226}0ёØ) --- p.86
Chapter 2.3 --- Methods
Chapter 2.3.1 --- Endotoxin Assay --- p.87
Chapter 2.3.2 --- Whole Blood Assay (WBA) --- p.88
Chapter 2.3.3 --- Isolation and Preparation of Plasma and Peripheral Blood Mononuclear Cells (PBMC) from EDTA Blood --- p.88
Chapter 2.3.4 --- Total RNA extraction --- p.89
Chapter 2.3.5 --- Flow Cytometric Analysis of T/B/NK Cell Ratios --- p.90
Chapter 2.3.6 --- Immunoassays of Plasma Samples or Culture Supernatant in WBA --- p.92
Chapter 2.3.6.1 --- Enzyme-linked Immunosorbent Assay (ELISA) --- p.92
Chapter 2.3.6.2 --- Human Thl/Th2 Cytokine Cytometric Bead Assay (CBA) --- p.93
Chapter 2.3.7 --- mRNA Expression Study --- p.94
Chapter 2.3.7.1 --- Agarose Gel Electrophoresis --- p.94
Chapter 2.3.7.2 --- cDNA Expression Array Analysis --- p.95
Chapter 2.3.8 --- Statistical Analysis --- p.96
Chapter CHAPTER 3: --- ENDOTOXIN LEVEL OF YUN ZHI-DANSHEN CAPSULES & SAFETY MEASURES ON STUDY POPULATION IN THE CLINICAL TRIAL
Chapter 3.1 --- Introduction --- p.98
Chapter 3.2 --- Results --- p.101
Chapter 3.2.1 --- Endotoxin Level of the Yun Zhi and Danshen Active Capsule --- p.101
Chapter 3.2.2 --- Study Population --- p.103
Chapter 3.2.3 --- Dropout Cases --- p.103
Chapter 3.2.4 --- Safety Parameters --- p.104
Chapter 3.2.5 --- Compliance Rates --- p.104
Chapter 3.3 --- Discussion --- p.109
Chapter CHAPTER 4: --- FLOW CYTOMETRIC ANALYSIS OF T/B/NK CELL RATIOS OF HEALTHY SUBJECTS TAKING YUN ZHI-DANSHEN CAPSULES
Chapter 4.1 --- Introduction --- p.112
Chapter 4.2 --- Results --- p.118
Chapter 4.2.1 --- The Percentage and Absolute Count of T Lymphocytes (CD3+) --- p.118
Chapter 4.2.2 --- The Percentage and Absolute Count of T Helper (TH) Lymphocytes (CD3+ CD4+) --- p.121
Chapter 4.2.3 --- The Percentage and Absolute Count of Cytotoxic T (CTL) and T Suppressor (Ts) Lymphocytes (CD3+ CD8+) --- p.124
Chapter 4.2.4 --- The Ratio of T Helper Lymphocytes (CD3+ CD4+) and Cytotoxic T (CTL) and T Suppressor (Ts) Lymphocyes (CD3+ CD8+) --- p.127
Chapter 4.2.5 --- The Percentage and Absolute Count of B Lymphocytes (CD19+) --- p.129
Chapter 4.2.6 --- The Percentage and Absolute Count of NK Lymphocytes (CD3- CD 16+ and/or CD56+) --- p.132
Chapter 4.2.7 --- The Absolute Count of Lymphocytes (CD45+) --- p.135
Chapter 4.3 --- Discussion --- p.138
Chapter CHAPTER 5: --- PLASMA CONCENTRATION OF SOLUBLE CYTOKINE RECEPTOR AND EX VIVO CYTOKINE PRODUCTION OF HEALTHY SUBJECTS TAKING YUN ZHI-DANSHEN CAPSULES
Chapter 5.1 --- Introduction --- p.142
Chapter 5.2 --- Results --- p.147
Chapter 5.2.1 --- Plasma Concentration of Soluble IL-2 Receptor --- p.147
Chapter 5.2.2 --- Ex vivo Cytokine Production --- p.147
Chapter 5.2.3 --- Mitogen Induced IL-6 Production --- p.150
Chapter 5.2.4 --- Mitogen Induced IFN- γ Production --- p.150
Chapter 5.2.5 --- Mitogen Induced TNF- a Production --- p.153
Chapter 5.2.6 --- Mitogen Induced IL-10 Production --- p.153
Chapter 5.3 --- Discussion --- p.156
Chapter CHAPTER 6: --- "GENE EXPRESSION OF CYTOKINES, CHEMOKINES AND RECEPTORS OF PBMC OF HEALTHY SUBJECTS TAKING YUN ZHI- DANSHEN CAPSULES"
Chapter 6.1 --- Introduction --- p.162
Chapter 6.2 --- Results --- p.165
Chapter 6.2.1 --- Gene Expression of IL-2 Receptor β chain --- p.165
Chapter 6.2.2 --- Gene Expression of IL-2 Receptor γ chain --- p.165
Chapter 6.2.3 --- Gene Expression of IL-6 Receptor --- p.166
Chapter 6.2.4 --- "Gene Expression of Other Cytokines, Chemokines and Receptors" --- p.169
Chapter 6.3 --- Discussion --- p.172
Chapter CHAPTER 7: --- CONCLUDING REMARKS AND FUTURE
PERSPECTIVES --- p.176
APPENDICES --- p.184
REFERENCES --- p.192

In a mouse model of pulmonary thromboembolism induced by a collagen-adrenaline mixture, the SFE extract and ligustilide reduced the paralysis-death ratio, and the anti-thrombotic response of senkyunolide A was more pronounced. The effect of BDPH was not significant. Neither the SFE extract nor the three phthalides prolonged bleeding time in tail-transected mice.
In a rat myocardial ischemia-reperfusion model involving coronary artery ligation, 7-day pre-treatment with the SFE extract and ligustilide reduced ventricular arrhythmias in isolated hearts. BDPH and senkyunolide A were without significant effects.
In rat platelet-rich plasma, platelet aggregation induced by collagen and U46619 but not by adenosine diphosphate was inhibited by the SFE extract. Ligustilide inhibited the responses of all three agonists, while BDPH and senkyunolide A inhibited the collagen response only.
Raw Rhizoma Chuanxiong herb and its crude extract as obtained by supercritical fluid extraction (SFE) comprised mainly phthalides. The SFE extract and three representative phthalides, butylidenephthalide (BDPH), ligustilide and senkyunolide A, were studied on vasorelaxation, myocardial ischemia, platelet aggregation and thrombosis. The mechanisms underlying BDPH-mediated vasorelaxation were also explored.
Rhizoma Chuanxiong, the dried rhizome of Ligusticum chuanxiong Hort., is a common traditional Chinese medicine used for the treatment of cardiovascular diseases. Surprisingly, the scientific basis of its action is poorly understood. The current study aims to establish the pharmacological basis of the cardiovascular effects of Rhizoma Chuanxiong and its active constituents by examining their effects in several cardiovascular domains.
The current study demonstrated various cardiovascular actions of Rhizoma Chuanxiong, and thereby established the pharmacological basis of the effects of the herb. Phthalides, in particular BDPH, ligustilide and senkyunolide A, were important contributors to such actions. Future investigation of the SFE extract and/or individual phthalides related to the progression from in vitro and in vivo effectiveness to clinical efficacy is much anticipated.
The SFE extract, BDPH, ligustilide and senkyunolide A produced vasorelaxation on isolated preparations of rat aorta, rat saphenous vein and pig coronary artery. BDPH-mediated relaxation appeared to involve both extracellular Ca 2+-dependent (L-type voltage-operated, receptor-operated and store-operated Ca2+ channels) and independent (NO modulation, Ca2+ release from internal stores and Ca2+ desensitization) mechanisms. BDPH was also observed to augment relaxation induced by sodium nitroprusside and forskolin through mechanisms that remain undefined.
Chan Sun Kin Sunny.
"July 2005."
Advisers: G. Lin; R. L. Jones.
Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1575.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2005.
Includes bibliographical references (p. 190-209).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstracts in English and Chinese.
School code: 1307.

Conclusion 1. The results of the research partly supported hypothesis 1a. There was a significant reduction in both endothelium-dependent arterial relaxation following a surgical menopause. The results of the research partly supported hypothesis 1b. There was a significant reduction in endothelium-dependent arterial relaxation but no significant effect on endothelium-independent arterial relaxation.
Conclusion 2. The results of the research partly supported hypothesis 2a. The addition of unopposed oestrogen significantly improved endothelium-dependent but not endothelium-independent arterial relaxation. The results of the research supported hypothesis 2b. The addition of oestradiol combined with progestogen (norethisterone acetate) reversed the reduction in arterial relaxation caused by a surgical menopause. The results of the research partly supported hypothesis 2c. The addition of tibolone reversed the reduction endothelium-dependent but not endothelium-independent arterial relaxation. The results of the research partly supported hypothesis 2d. The addition of oestradiol combined with a progestogen (norethisterone acetate) reversed the reduction in endothelium-dependent but not endothelium-independent arterial relaxation.
Conclusion 3. The results of the research partly supported hypothesis 3a. Endothelium-dependent arterial relaxation but no endothelium-independent arterial relaxation was improved after the addition of menopausal hormone therapy using oestrogen combined with a progestogen in a continuous manner. The results of the research did not support hypothesis 3b. Neither endothelium-dependent arterial relaxation nor the endothelium-independent arterial relaxation was improved by cyclical menopausal HT.
Conclusion 4. The results of the research did not support hypothesis 4. The addition of menopausal hormone therapy using combined oestrogen with progestogen did not improve arterial relaxation in postmenopausal women with established coronary heart disease.
Hypothesis 2. This hypothesis examined three different types of commonly used menopausal HT. That unopposed oestrogen (2a), oestrogen combined with a progestogen (2b and 2d) or a synthetic steriod that has oestrogenic, progestogenic as well as androgenic activity (tibolone, 2c), reverse the reduction in arterial relaxation following menopause in Hong Kong Chinese women.
Hypothesis 3. That menopausal hormone therapy using oestrogen combined with progestogen given in either continuous (3a) or cyclical (3b) regimens improves arterial relaxation in postmenopausal Hong Kong Chinese women.
Hypothesis 4. That menopausal hormone therapy using combined oestrogen with progestogen improves arterial relaxation in postmenopausal Hong Kong Chinese women with established coronary heart disease.
Menopausal HT can in general at least partially reverse changes in arterial relaxation in postmenopausal women. Different types of menopausal HT exhibit different effects on arterial relaxation. In healthy vessels, menopause HT mainly reverses the endothelium-dependent vascular effect, but it remains unclear how menopausal HT affects the endothelium-independent vascular effect. However, with established coronary heart disease, menopausal HT cannot reverse the changes in vascular reactivity.
Summary. Menopause results in a reduction in arterial relaxation. However, GnRHa temporarily induced menopause in young women, the endothelium-independent vasodilatation was not impaired. This difference can be partly explained by the difference in age as vascular reactivity is age dependent. Secondly, GnRHa works with an initial phase of increase in oestrogen production resulting in a shorter duration of hypo-oestrogenism resulting in the lack of impairment on endothelium-independent vasodilatation.
This thesis tested the following hypotheses: Hypothesis 1. That vascular reactivity decreases after the menopause as shown in premenopausal Hong Kong Chinese women with either a surgical (1a) or a medically induced (1b) menopause.
This thesis will examine the effects of menopause and menopausal HT on arterial reactivity which is an indirect measurement of vascular function. Previous studies have shown that oestrogen is a potent coronary artery vasodilator, and this effect may be mediated via both endothelium-dependent and endothelium-independent mechanisms. One method of assessing vascular reactivity is to use ultrasound measurement of changes in brachial artery diameter in response to certain stimuli. Using this technique, changes in both endothelium-dependent and endothelium-independent vasodilatation can be measured. Increased rather than decreased arterial relaxation after stimulus can be viewed as a favourable response.
Yim, So-fan.
Adviser: C. J. Haines.
Source: Dissertation Abstracts International, Volume: 68-09, Section: B, page: 5873.
Thesis (M.D.)--Chinese University of Hong Kong, 2006.
Includes bibliographical references (p. 159-194).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
School code: 1307.

by Fung Yin Lee, Annie.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1995.
Includes bibliographical references (leaves 181-189).
ABSTRACT --- p.i
LIST OF ABBREVIATIONS --- p.iv
ACKNOWLEDGEMENT --- p.v
TABLE OF CONTENTS --- p.vi
LIST OF FIGURES --- p.ix
INTRODUCTION --- p.1
LITERATURE REVIEW --- p.4
Chapter I. --- A. Arterial pressure --- p.4
Chapter B. --- Regulation of arterial pressure --- p.7
Chapter II. --- Hypertension --- p.14
Chapter III. --- Treatment of hypertension --- p.29
Chapter IV. --- Plants and their effects on blood pressure --- p.48
Chapter V. --- Characteristics of the three plants being studied --- p.50
MATERIALS AND METHODS --- p.55
Chapter A. --- Preparative procedures --- p.55
Chapter 1. --- Preparation of plant extracts --- p.55
Chapter 2. --- Animal preparation for invivo blood pressure measurement --- p.56
Chapter 3. --- Preparation of right atria for in vitro studies --- p.56
Chapter 4. --- Preparation of artery strips for in vitro studies --- p.57
Chapter 5. --- Preparation for diuretic studies --- p.58
Chapter B. --- Experiments done --- p.60
Chapter 1. --- Cumulative dose response of individual plant extract --- p.60
Chapter 2. --- Combination of plant extracts --- p.60
Chapter 3. --- Pharmacological antagonists studies --- p.64
Chapter a. --- Autonomic ganglion transmission --- p.64
Chapter b. --- Alpha adrenergic activity --- p.64
Chapter c. --- Beta adrenergic activity --- p.65
Chapter d. --- Cholinergic activity --- p.65
Chapter e. --- Histaminergic activity --- p.65
Chapter f. --- Serotoninergic activity --- p.65
Chapter 4. --- Urinary and sodium excretionin water loaded rats --- p.66
Chapter 5. --- Studies on chronotropic and inotropic effects on isolated right atrium --- p.66
Chapter a. --- Effect of individual plant extract --- p.66
Chapter b. --- Effect of combination of plant extracts --- p.66
Chapter 6. --- Effect of plant extract on contractile responses of rat tail artery strips --- p.70
Chapter a. --- Effect of individual plant extract --- p.70
Chapter b. --- Effect of combination of plant extracts --- p.70
Chapter 7. --- Effect of acute oral feeding of plant extracts on blood pressure of rats --- p.71
Chapter C. --- Statistics --- p.71
RESULTS
Chapter A. --- Preparation of plant extracts --- p.72
Chapter B. --- Effect of plant extracts on blood pressure changes --- p.72
Chapter 1. --- Individual plant extract --- p.72
Chapter 2. --- Combination of two plant extracts --- p.73
Chapter 3. --- Combination of three plant extracts --- p.76
Chapter C. --- Pharmacological antagonist studies --- p.79
Chapter 1. --- Autonomic ganglion transmission --- p.79
Chapter 2. --- Alpha adrenergic activity --- p.79
Chapter 3. --- Beta adrenergic activity --- p.81
Chapter 4. --- Cholinergic activity --- p.82
Chapter 5. --- Histaminergic activity --- p.83
Chapter 6. --- Serotoninergic activity --- p.84
Chapter D. --- Urinary and sodium excretion in water loaded rats --- p.85
Chapter E. --- Chronotropic and inotropic studies of isolated right atrium --- p.88
Chapter 1. --- Effect of individual plant extract --- p.88
Chapter 2. --- Effect of combination of plant extracts --- p.89
Chapter F. --- Effect of plant extracts on contractile responses of rat tail artery strips --- p.101
Chapter G. --- Effect of acute oral feeding of plant extracts on MAP of rats --- p.102
DISCUSSION --- p.156
Chapter A. --- Comment on preparation of plant extracts --- p.156
Chapter B. --- The hypotensive effects of the plant extracts --- p.157
Chapter C. --- The mechanism of action --- p.159
Chapter D. --- The renal effect of plant extracts --- p.161
Chapter E. --- The interaction of the hypotensive effect of plant extracts --- p.164
Chapter F. --- In vitro studies --- p.167
Chapter G. --- The oral effect of the plant extracts --- p.174
SUMMARY --- p.176
CONCLUSION --- p.179
REFERENCES --- p.181
APPENDIX --- p.190
"Appendix I To study the hypotensive effects of trypsin treated green bean, rue and kelp" --- p.191
"Appendix II To study the hypotensive effects of ether treated green bean, rue and kelp" --- p.194

Yuen Yee Man.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2005.
Includes bibliographical references (leaves 108-125).
Abstracts in English and Chinese.
TITLE PAGE --- p.1
ACKNOWLEGDEMENTS --- p.2
ABSTRACT --- p.3
摘要 --- p.5
table of contents --- p.7
list of figures and tables --- p.13
CHAPTER 1 GENERAL INTRODUCTION --- p.16
Chapter 1.1 --- role of PHYTOESTROGENS in soy and red WINE the PREVENTION OF CARDIOVASCULAR DISEASES (CVD) --- p.17
Chapter 1.1.1 --- INTRoduction and Classification of Phytoestrogens --- p.17
Chapter 1.1.2 --- estrogenic1ty of phytoestrogens and theIr abundancesin Plasma --- p.18
Chapter 1.1.3 --- phytoestrogens as one of the active components In cvd Protection --- p.21
Chapter 1.1.4 --- effects of Phytoestrogens on LDL Receptor and Apolipoprotein A-1 --- p.22
Chapter 1.2 --- role of estrogen receptors (ers) in gene regulation --- p.24
Chapter 1.2.1 --- "structure, Classification and tissue distribution of ERS" --- p.24
Chapter 1.2.2 --- ligands for ERS --- p.25
Chapter 1.2.3 --- mechaniSMS OF LIgands-ERS complex in GENE regulation --- p.27
Chapter 1.2.4 --- ligand-independent ER activation --- p.28
Chapter 1.3 --- aims and scopes of investigation --- p.29
Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.30
Chapter 2.1 --- chemicals and materials --- p.30
Chapter 2.1.1 --- Chemicals --- p.30
Chapter 2.1.2 --- Plasmids --- p.30
Chapter 2.2 --- mammalian cell culture maintainence --- p.30
Chapter 2.2.1 --- Maintenance of Cells --- p.31
Chapter 2.2.2 --- Preparation of Cell Stock --- p.31
Chapter 2.2.3 --- Cell Recovery from Liquid Nitrogen Stock --- p.31
Chapter 2.3 --- manipulation of dna --- p.31
Chapter 2.3.1 --- isolation of HEPG2 cells genonmic DNA --- p.31
Chapter 2.3.2 --- separation and purification of dna from agarose gel --- p.31
Chapter 2.3.3 --- Restriction digestionof DNA --- p.32
Chapter 2.3.4 --- Ligation of DNA Fragments --- p.32
Chapter 2.3.5 --- Transformation of --- p.32
Chapter 2.3.6 --- Small Scale Plasmids Purification from DH5a --- p.32
Chapter 2.4 --- construction of expression and reporter plasmids --- p.33
Chapter 2.4.1 --- Construction of Estrogen Receptorα (Erα) Expression Vectors --- p.33
Chapter 2.4.2 --- construction of reporter vectors of LDLR promoter and the Respective Mutants --- p.33
Chapter 2.4.3 --- Construction of Reporter Vectors of APOAI Promoter and the Respective Mutants --- p.33
Chapter 2.5 --- determination of promoter transcrtiption activities --- p.34
Chapter 2.5.1 --- Transient Transfection of Cell with ERa Expression Vector and Promoter Reporter using Lipofectamine PLUS Reagent --- p.34
Chapter 2.5.2 --- Dual Luciferase Assay --- p.34
Chapter 2.6 --- semi-quantitative and quantitative rt-pcr assay --- p.34
Chapter 2.6.1 --- Transient transfection of Cell with ERa Expression Vector Using Lipofectamine PLUS Reagent --- p.34
Chapter 2.6.2 --- "Isolation of RNA using TRIzol® Reagent (Life Technology, USA)" --- p.35
Chapter 2.6.3 --- Quantitation of RNA --- p.35
Chapter 2.6.4 --- First Strand cDNA Synthesis --- p.35
Chapter 2.6.5 --- Sem卜Quantitative PCR Reactions --- p.35
Chapter 2.6.6 --- Quantitative PCR Reactions --- p.36
Chapter 2.7 --- western blotting analysis --- p.36
Chapter 2.8 --- statistical methods --- p.36
Chapter CHAPTER 3 --- REGULATION BY PHYSIOLOGICAL LEVEL OF 17B-ESTRADIOL ON APOLIPOPROTEIN A-I AND LOW-DENSITY- LIPOPROTEIN RECEPTOR IN HEPG2 CELLS --- p.37
Chapter 3.1 --- introduction --- p.37
Chapter 3.2 --- results --- p.39
Chapter 3.2.1 --- Determination of transient transfection functionality of estrogen receptors in hepg2 cells --- p.39
Chapter 3.2.2 --- Effect of 17β-Estradiolon LDLR promoter transcription activity --- p.39
Chapter 3.2.3 --- Effect of 17β-Estradiol on apoai promoter transcription activity --- p.40
Chapter 3.2 --- discussion --- p.47
Chapter CHAPTER 4 --- SOY ISOFLAVONES AND RESVERATROL DISPLAY DIFFERENT MECHANISM IN THE UP-REGULATION OF LOVV-DENSITY-LIPOPROTEIN RECEPTOR IN HEPG2 CELLS --- p.49
Chapter 4.1 --- introduction --- p.49
Chapter 4.2 --- results --- p.54
Chapter 4.2.1 --- Association of ERα and isoflavones or resveratrol on LDLR promoter transcription activity --- p.54
Chapter 4.2.2 --- Association of ERβ and isoflavones or resveratrol on LDLR promoter transcription activity --- p.54
Chapter 4.2.3 --- "Role of MAP Kinase, PKA and PKC in isoflavones and resveratrol induced LDLR promoter transcription" --- p.55
Chapter 4.2.4 --- Identification of promoter regions responsible for induction of LDLR transcription by isoflavones in the presence OF ERα --- p.55
Chapter 4.2.5 --- Identification of promoter regions responsible for induction of LDLR TRANSCRIPTION BY resveratrol IN THE ABSENCE OF ERα --- p.56
Chapter 4.3 --- DISCUSSION --- p.75
Chapter CHAPTER 5 --- SOY ISOFLAVONES AND RESVERATROL UP-REGULATE APOLIPOPROTEIN A-I SIMILAR TO 17B-ESTRADIOL IN HEPG2 CELLS --- p.80
Chapter 5.1 --- INTRODUCTION --- p.80
Chapter 5.2 --- RESULTS --- p.84
Chapter 5.2.1 --- Association of ERα phytoestrogens on APCAI gene expression --- p.84
Chapter 5.2.2 --- Association of ERβ and isoflavones or resveratrol on APOAI promoter transcription activity --- p.85
Chapter 5.2.3 --- "Role of MAP Kinase, PKA and PKC in isoflavones and resveratrol in APOAI promoter transcription in the presence of ERα" --- p.85
Chapter 5.2.4 --- Identification of promoter regions responsible for induction of APOAI transcription by isoflavones and resveratrol in the presence of ERα --- p.85
Chapter 5.3 --- DISCUSSION --- p.100
Chapter CHAPTER 6 --- GENERAL DISCUSSION --- p.103
Chapter CHAPTER 7 --- SUMMARY --- p.106
BIBLIOGRAPHY --- p.108
APPENDIX 1 ABBREVIATIONS --- p.126
APPENDIX 2 MATERIALS AND METHODS --- p.129
APPENDIX 3 PRIMER LISTS --- p.145
APPENDIX 4 REAGENTS AND BUFFERS --- p.147

Background Iron deficiency (ID) is the world‟s most prevalent micronutrient deficiency and predominantly affects developing countries, also South Africa. In areas with low fish consumption and high n-6 PUFA vegetable oil intake, there is a risk for having inadequate n-3 PUFA status. Both iron and n-3 PUFA play important roles in the immune response, and supplementation is a strategy to alleviate deficiencies. However, little is known about potential interactive effects between concurrent iron and n-3 PUFA supplementation on the immune system. This is also important in the context that iron supplementation may be unsafe and may increase morbidity and mortality. Aim The overall aim of this thesis was to assess the effects of iron and docosahexaenoic (DHA)/eicosapentaenoic acid (EPA) supplementation, alone and in combination, on the immune system of ID children. More specifically, these effects were investigated on the occurrence and duration of illness and school-absenteeism due to illness, peripheral blood mononuclear cell (PBMC), red blood cell (RBC) and plasma total phospholipid fatty acid composition, iron status, fatty acid-derived immune modulators and targeted PBMC gene expression. Furthermore, association of PBMC, RBC and plasma total phospholipid fatty acid composition with allergic disease, were also examined. Design In a 2-by-2 factorial, randomised, double-blind, placebo-controlled trial, South African children (n = 321, aged 6–11 y) were randomly assigned to receive oral supplements of either 1) iron (50 mg as ferrous sulphate) plus placebo; 2) DHA/EPA (420/80 mg) plus placebo; 3) iron plus DHA/EPA (420/80 mg); or 4) placebo plus placebo for 8.5 mo, four times per week. Absenteeism and illness symptoms were recorded and biochemical parameters for compliance as well as parameters fundamental to immune function were assessed at baseline and endpoint. Furthermore, in a cross-sectional design, associations of allergic disease with baseline fatty acid composition of PBMC, RBC and plasma were examined. Results The combination of iron and DHA/EPA significantly attenuated respiratory illness caused by iron supplementation. DHA/EPA supplementation alone improved respiratory symptoms at school, but increased headache-related absenteeism. DHA/EPA and iron supplementation individually tended to increase and decrease anti-inflammatory DHA and EPA-derived mediators, respectively. Furthermore the anti-inflammatory DHA-derived immune mediator, 17HDHA was higher in the DHA/EPA plus placebo and iron plus DHA/EPA groups than in the iron plus placebo group. Also, the pro-inflammatory arachidonic acid (AA)-derived modulators (5- and 15-hydroxyeicosapentaenoic acid) were significantly lower in the iron plus DHA/EPA group compared to the placebo plus placebo groups. In the study population, 27.2% of the children had allergic disease and AA in PBMC phospholipids was significantly lower in the allergic children than in the non-allergic children. In RBC phospholipids dihomo-gamma-linolenic acid (DGLA) and the ratio of DGLA: linoleic acid (LA) correlated negatively and the n-6:n-3 PUFA ratio positively with total immunoglobulin E (tIgE). Furthermore, trans-C18:1n-9, tended to be higher in the allergic group. Conclusion DHA/EPA prevented respiratory illness caused by iron supplementation and although DHA/EPA on its own reduced respiratory morbidity when the children were present at school, surprisingly it increased the likelihood of being absent with headache and fever. The biochemical findings compliment the clinical results and support previous observations about DHA/EPA supplementation to reduce inflammation, but add to the current knowledge base that a relatively high oral dose of non-haem iron modulates circulating lipid-derived immune modulators and related gene expression. Furthermore, when supplementing with iron and DHA/EPA combined, in this ID population with low fish intake, the anti-inflammatory effect of DHA/EPA is maintained concurrently with attenuation of respiratory morbidity. This finding support the notion that excess iron (probably as non-transferrin bound iron) becomes available for pathogens and is probably why we found that iron increased respiratory infectious morbidity. The improved clinical outcome with combined supplementation seems to be related to increased lipid-mediator synthesis gene expression and the availability of DHA/EPA, leading to a more pro-resolving profile and enhanced immune competence. Overall these results give better insight into immune function and infectious morbidity in relation to n-3 PUFA and iron status and treatment, as well as the possible association of fatty acid status with allergic disease in young South-African school children.
PhD (Nutrition), North-West University, Potchefstroom Campus, 2015

PURPOSE: Cytochrome P450 2W1 (CYP2W1) is a monooxygenase detected in 30% of colon cancers, whereas its expression in nontransformed adult tissues is absent, rendering it a tumor-specific drug target for development of novel colon cancer chemotherapy. Previously, we have identified duocarmycin synthetic derivatives as CYP2W1 substrates. In this study, we investigated whether two of these compounds, ICT2705 and ICT2706, could be activated by CYP2W1 into potent antitumor agents. EXPERIMENTAL DESIGN: The cytotoxic activity of ICT2705 and ICT2706 in vitro was tested in colon cancer cell lines expressing CYP2W1, and in vivo studies with ICT2706 were conducted on severe combined immunodeficient mice bearing CYP2W1-positive colon cancer xenografts. RESULTS: Cells expressing CYP2W1 suffer rapid loss of viability following treatment with ICT2705 and ICT2706, whereas the CYP2W1-positive human colon cancer xenografts display arrested growth in the mice treated with ICT2706. The specific cytotoxic metabolite generated by CYP2W1 metabolism of ICT2706 was identified in vitro. The cytotoxic events were accompanied by an accumulation of phosphorylated H2A.X histone, indicating DNA damage as a mechanism for cancer cell toxicity. This cytotoxic effect is most likely propagated by a bystander killing mechanism shown in colon cancer cells. Pharmacokinetic analysis of ICT2706 in mice identified higher concentration of the compound in tumor than in plasma, indicating preferential accumulation of drug in the target tissue. CONCLUSION: Our findings suggest a novel approach for treatment of colon cancer that uses a locoregional activation of systemically inactive prodrug by the tumor-specific activator enzyme CYP2W1.