Chan Chun-pong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 171-176). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 摘要 --- p.iii / Abbreviation --- p.iv / Table of contents --- p.v / List of figures --- p.xi / List of tables --- p.xvi / Chapter Chapter 1 --- introduction / Chapter 1.1 --- Traditional Chinese medicines (TCMs) --- p.2 / Chapter 1.2 --- Liver disorders in Asia Pacific region --- p.3 / Chapter 1.3 --- Classification of liver disorders --- p.7 / Chapter 1.3.1 --- Hepatitis --- p.8 / Chapter 1.3.1.1 --- Hepatitis A virus infection --- p.8 / Chapter 1.3.1.2 --- Hepatitis B virus infection --- p.9 / Chapter 1.3.1.3 --- Hepatitis C virus infection --- p.11 / Chapter 1.3.1.4 --- Hepatitis D virus infection --- p.12 / Chapter 1.3.1.5 --- Hepatitis E virus infection --- p.12 / Chapter 1.3.2 --- Cancer of the liver --- p.13 / Chapter 1.3.2.1 --- Hepatocellular carcinoma --- p.13 / Chapter 1.3.2.2 --- Cholangiocarcinoma --- p.14 / Chapter 1.3.2.3 --- Metastatic liver cancer --- p.14 / Chapter 1.4 --- Conventional treatment of liver disorders --- p.14 / Chapter 1.5 --- Role of traditional Chinese medicines in hepatoprotective functions --- p.16 / Chapter 1.5.1 --- Abri Herba (Abrus Cantoniensis Hance) --- p.17 / Chapter 1.5.2 --- Rhizoma Coptidis (Coptidis chinensis Franch) --- p.18 / Chapter 1.5.3 --- Fructus Forsythia (Forsythia suspense (Thunb) Vahl) --- p.22 / Chapter 1.6 --- Molecular studies of hepatoprotective effects of TCMs --- p.26 / Chapter 1.6.1 --- Roles of detoxofication enzymes in hepatoprotection --- p.27 / Chapter 1.6.2 --- Studies of growth-related genes in cell cycle control --- p.29 / Chapter 1.7 --- Aims of project --- p.32 / Chapter 1.8 --- Application of the project --- p.33 / Chapter Chapter 2 --- Methods and materials --- p.34 / Chapter 2.1 --- Screening of traditional Chinese medicines --- p.35 / Chapter 2.2 --- Preparation of TCMs --- p.35 / Chapter 2.2.1 --- Preparation of aqueous extracts of TCMs --- p.35 / Chapter 2.2.2 --- Preparation of active components of TCMs --- p.36 / Chapter 2.3 --- In vitro assays --- p.40 / Chapter 2.3.1 --- Cell culture --- p.40 / Chapter 2.3.2 --- Cytotoxicity test --- p.40 / Chapter 2.4 --- Screening of expressed gene induced by TCMs --- p.41 / Chapter 2.4.1 --- RNA preparation --- p.41 / Chapter 2.4.2 --- cDNA array hybridization --- p.42 / Chapter 2.4.3 --- Reverse Transcription --- p.43 / Chapter 2.5 --- Confirmation of expressed genes induced by TCMs --- p.44 / Chapter 2.5.1 --- Semi-quantitative PCR analysis --- p.44 / Chapter 2.5.2 --- Northern blot analysis --- p.46 / Chapter 2.6 --- Studies of effects of TCMs in gene expression --- p.47 / Chapter 2.6.1 --- Dosage-course study --- p.47 / Chapter 2.6.2 --- Time-course study --- p.48 / Chapter Chapter 3 --- Results --- p.50 / Chapter 3.1 --- "Cytotoxicity test of A.H., R.C. and F.F" --- p.51 / Chapter 3.2 --- "Molecular screening of expressed gene induced by A.H., R.C., F.F" --- p.58 / Chapter 3.3 --- Confirmation of expressed gene using semi-quantitative RT- PCR --- p.70 / Chapter 3.3.1 --- Dosage-course and time-course studies of A.H. using RT- PCR --- p.70 / Chapter 3.3.2 --- Dosage-course and time-course studies of R.C. using RT- PCR --- p.94 / Chapter 3.3.3 --- Dosage-course and time-course studies of A.H. using RT- PCR --- p.113 / Chapter 3.4 --- Confirmation of expressed gene using northern blot anaylsis --- p.118 / Chapter 3.4.1 --- Dosage-course and time-course studies of effects of A.H. and L- abrine in Northern blot analysis --- p.118 / Chapter 3.4.2 --- Dosage-course and time-course studies of effects of R.C. and berberine in Northern blot analysis --- p.129 / Chapter 3.4.3 --- Dosage-course and time-course studies of effects of F.Fin Northern blot analysis --- p.147 / Chapter Chapter 4 --- Discussion --- p.152 / Chapter 4.1 --- "Roles of A.H., R.C. and F.F. in treatment and prevention of liver disorders" --- p.153 / Chapter 4.2 --- "Cytotoxicity effect A.H., R.C., and F.F. in liver cells" --- p.153 / Chapter 4.3 --- Effects of herbal medicines on the transcription of mRNA in liver cells --- p.155 / Chapter 4.3.1 --- Effects of treatment of A.H. in liver at transcriptional level … --- p.155 / Chapter 4.3.2 --- Effects of treatment of R.C. in liver at transcriptional level … --- p.156 / Chapter 4.3.3 --- Effects of treatment of R.C. in liver at transcriptional level --- p.157 / Chapter 4.4 --- Comparison of results of RT-PCR and Northern blot analysis --- p.157 / Chapter 4.4.1 --- Comparison of the effects of time and dosage-course studies of DTD expression induced by A.H. and L-abrine --- p.157 / Chapter 4.4.2 --- Comparison of the effects of time and dosage-course studies of p21;cip;waf1 expression induced by A.H. and L-abrine --- p.158 / Chapter 4.4.3 --- Comparison of the effects of time and dosage-course studies of c-myc responsive protein; rcl expression induced by R.C. and berberine --- p.159 / Chapter 4.4.4 --- Comparison of the effects of time and dosage-course studies of GST Ya expression induced by R.C. and berberine --- p.160 / Chapter 4.4.5 --- Comparison of the effects of time and dosage-course studies of GST 7-7 expression induced by F.F --- p.160 / Chapter 4.5 --- Biochemical significance of genes induced by hepatoprotective TCMs --- p.161 / Chapter 4.5.1 --- Roles of significant expression of detoxifying enzymes induced by TCMs in liver cells --- p.161 / Chapter 4.5.2 --- Roles of induction of growth-related c-myc responsive protein; rcl in R.C. treated liver cells --- p.167 / Chapter 4.5.3 --- Roles of increased p21;cip;waf1 expression in A.H. treated liver cells --- p.168 / Chapter 4.6 --- Conclusion --- p.169

引發獲得性免疫缺陷綜合癥（AIDS）的人類免疫缺陷病毒（HIV）是一種逆轉錄病毒。過去的十餘年間，高效抗逆轉錄病毒治療療法（HARRT），在抗病毒感染方面取得了很大的成功。高效抗逆轉錄病毒治療療法是一種將多種抗逆轉錄病毒藥物複合的藥物聯用療法。然而，因為病毒的逆轉錄過程極易突變，導致HIV已經可以對大多數使用的抑製藥物產生抗藥性。因此，有越來越多的需要去尋找新型的抗病毒複製機理，例如將人體細胞蛋白作為載體，來達到克服病毒抗藥性的目的。 / HIV-1的複製離不開宿主細胞的剪接因子，例如SR蛋白。選擇性剪接因子ASF/SF2，一個典型的調控pre-RNA剪接的SR蛋白，在HIV-1的pre-mRNA剪接和複製中起到了很重要的調控作用。ASF/SF2和其他SR蛋白一樣，都被丝氨酸/苏氨酸蛋白激酶（SRPK）磷酸化，磷酸化位點位於C端的丝氨酸/苏氨酸結構域（RS domain）。SRPK通過磷酸化來調節ASF/SF2在細胞中的分佈。對於SRPK 和ASF/SF2複合物的結構學和功能學研究指出，ASF/SF2的docking motif和SRPK1的遠離活性位點的docking groove存在很強的相互作用。而這種相互作用是調節磷酸化過程關鍵。所以，在我們的研究過程中，我們希望通過阻斷2個蛋白的相互作用來干擾ASF/SF2的磷酸化，進而抑制其在HIV-1 pre-mRNA剪接過程中的活性。 / 我們採用以結構為基礎的藥物模擬篩選，來選擇潛在的抑制物，達到通過抑制物與docking groove的相互作用來阻斷ASF/SF2和SRPK1的相互作用，以達到抑制磷酸化的目的。我們使用的數據庫來自于ZINC數據庫（UCSF），包括天然產物數據庫和SPECS。我們採用AutoDock Vina 和AutoDock 4.2 二個模擬軟件來栓選數據庫中351473个化合物。并從中選出50個潛在的化合物用作之後的化學生物學測試。體外的激酶活性試驗顯示，6個化合物對ASF/SF2的磷酸化有抑製作用。 / 體外的HIV-1 pre-mRNA剪接實驗顯示，5個化合物在逆轉錄PCR（RT-PCR）中有一定得抑制效果。和DMSO對照組相比，在抑製劑作用下剪接產物的生成被抑制。HIV-1病毒合胞體感染實驗顯示，有一個化合物對病毒的感染起到了一定的抑制作用。 / 其他的測試實驗還在進行中，包括對SRPK1和抑制物複合物的結構研究，從而更好的研究抑制物的作用機理。以及，採用表面等離子共振波譜來進行動力學研究和其他關於化合物在病毒複製過程中的實驗測試。 / Human immunodeficient virus (HIV) is a retrovirus that cause acquired immunodeficiency syndrome (AIDS). Highly active antiretroviral therapy (HAART) is a treatment of HIV infection that uses combinations of antiretroviral drugs and has achieved great success in the past two decades. However, since the reverse transcription process of viral RNA is notoriously prone to error, HIV-1 can acquire resistance to nearly all known inhibitors and has started to develop resistance to HAART. Therefore, there is an ongoing search for new drugs with novel inhibitory mechanism such as targeting cellular proteins essential for HIV-1 replication to overcome drug resistance of the virus. / HIV-1 mRNA undergoes complex splicing and the expression of the integrated HIV-1 provirus is largely dependent on the host’s splicing machinery which assembly requires splicing factors such as serine-arginine rich proteins (SR proteins). Alternative splicing factor/splicing factor 2 (ASF/SF2), a prototypic SR protein that is essential for pre-mRNA splicing, has been shown to play critical roles during HIV-1 pre-mRNA splicing and replication. ASF/SF2, like other SR proteins, is phosphorylated by SR protein-specific kinases (SRPKs) at its C-terminal arginine/serine (RS) domain, which governs its localization and metabolism. Structural and functional studies of SRPK1 in complex with ASF/SF2 has revealed that a docking groove on SRPK1 that is distal to the active site interacts strongly with a docking motif and the RS domain of ASF/SF2, leading to high affinity binding as well as regulating the mechanism of phosphorylation. In this study, we propose that by blocking this interaction, we might interfere the phosphorylation of ASF/SF2 and inhibit its activity during splicing of HIV-1 pre-mRNA. / Structure-based in silico screening method is adopted to identify potential inhibitors that bind to the docking groove of SRPK1 to block the binding and phosphorylation of ASF/SF2. The compound libraries being used include the Natual Products Database and SPECS database from ZINC (UCSF). 351,473 compounds have been screened using the program Autodock Vina as well as Autodock 4.0. Until now 50 potential candidates of inhibitor have been selected for biochemical analyses. In vitro kinase assays showed that six compounds exhibit inhibitory activity against the phosphorylation of ASF/SF2. / To test the effect of the selected inhibitors on the splicing of HIV-1 mRNA, ex vivo splicing assay has been performed. Current results showed that the synthesis of splicing products extracted from drug-treated cells was less efficient when compared to untreated cells. Biological assays testing the inhibitory effects of the compounds on viral infection are currently underway. Our preliminary result suggested that one of the compounds could indeed inhibit HIV-1 viral infection. / Other biochemical and biological analyses including structural study of kinase-inhibitor complexes to understand the mode of inhibition; measurement of binding kinetics using surface plasmon resonance spectroscopy (SPR); and biological assays testing the inhibitory effects of the compounds on replication are underway. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Yu, Xiyao. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 95-107). / Abstracts also in Chinese. / Abstract --- p.I / 摘要 --- p.III / Acknowledgements --- p.V / TABLE OF CONTENTS --- p.VI / LIST OF FIGURES --- p.IX / LIST OF TABLES --- p.XI / Chapter Chapter I --- : Introduction --- p.1 / Chapter 1.1 --- HIV, HAART and HIV Drug Resistance --- p.2 / Chapter 1.2 --- HIV-1 alternative splicing mechanism --- p.9 / Chapter 1.3 --- SR Protein Family --- p.13 / Chapter 1.4 --- Functional roles of SR protein in HIV pre-mRNA splicing --- p.16 / Chapter 1.5 --- Phosphorylation States of SR Proteins --- p.18 / Chapter 1.6 --- SR protein Kinase --- p.20 / Chapter 1.7 --- Interaction between SRPK1 and ASF/SF2 --- p.23 / Chapter 1.8 --- IDC16 and SPRIN340 --- p.26 / Chapter 1.9 --- Structure-based drug screening --- p.27 / Chapter 1.10 --- AutoDock Suite --- p.29 / Chapter 1.11 --- Kinase-substrate interaction inhibitors --- p.30 / Chapter 1.12 --- Focus of study --- p.34 / Chapter Chapter II --- : Materials and Methods --- p.35 / Chapter 2.1 --- Materials --- p.36 / Chapter 2.1.1 --- Bacterial strain --- p.36 / Chapter 2.1.2 --- Antibodies --- p.36 / Chapter 2.1.3 --- Cell line --- p.36 / Chapter 2.1.4 --- Plasmid --- p.36 / Chapter 2.1.5 --- Reagents --- p.38 / Chapter 2.2 --- Expression and purification of Recombinant protein --- p.38 / Chapter 2.3 --- In silico screening of inhibitors --- p.44 / Chapter 2.4 --- Kinase Glo Assay --- p.45 / Chapter 2.5 --- In vitro kinase assay --- p.45 / Chapter 2.6 --- Cell Culture --- p.46 / Chapter 2.7 --- MTT Assay --- p.46 / Chapter 2.8 --- Immunocytochemistry --- p.47 / Chapter 2.9 --- Ex vivo splicing assay --- p.47 / Chapter 2.10 --- Surface plasmon resonance spectroscope --- p.48 / Chapter Chapter III --- : Results --- p.50 / Chapter 3.1 --- In silico screening of inhibitors --- p.51 / Chapter 3.2 --- Selected Compounds Inhibits SRPK1 in Vitro --- p.60 / Chapter 3.2.1 --- Protein purification --- p.60 / Chapter 3.2.2 --- Inhibits ASF/SF2 Phosphorylation by SRPK --- p.66 / Chapter 3.3 --- Surface Plasmon Resonance Binding Competition Assay --- p.76 / Chapter 3.4 --- Inhibitors Alters HIV-1 Alternative Splicing ex Vivo --- p.79 / Chapter 3.5 --- Cytotoxic effect of candidate compound on HeLa cells --- p.84 / Chapter 3.6 --- Nature compound alters ASF/SF2 localization --- p.86 / Chapter Chapter IV --- : Discussion and Conclusion --- p.89 / References --- p.95

Background. Hypertension carries a high risk of cardiovascular complications. Patient medication non-compliance has been identified to be an major factor for suboptimal blood pressure control in clinical practice. Different strategies have been proposed to improve patient medication compliance but their effects on clinical outcomes were inconsistent. Methods . A telephone survey was conducted to examine patient medication compliance with anti-hypertensive drugs in Hong Kong. I then established a Pharmacist-managed Compliance Clinic in a public out-patient setting and provided individualized patient education to non-compliant patients identified by physicians. A telephone follow-up was arranged at 4-week after intervention followed by a more in-depth reassessment on subsequent physician clinic visit day. The immediate endpoint was patient compliance rate. Intermediate endpoint was systolic and diastolic blood pressure control. Other outcome measures were control of other cardiovascular risk factors and level of healthcare resources utilization. / Conclusion. Pharmacist-managed Compliance Clinic is effective in improving patient medication compliance and has positive impact on clinical outcomes. (Abstract shortened by UMI.) / Results. A total of 853 patients were successfully contacted and completed the patient survey. According to our definition, 80.4% of patients interviewed were considered to be compliant. Factors associated with medication compliance included multiple drug therapy, presence of drug adverse effects, patient's awareness of preventive nature of medication, rapport between patient and physician, and full-time working status. A causal model was successfully established with latent factors identified for medication non-compliance. The factors included patient's functional status, provision of health advice and concern from physician, and patient's knowledge regarding reasons for drug taking. Another two hundreds hypertensive patients were followed at the Pharmacist-managed Compliance Clinic. On average, each patient attended 1.3 pharmacist visits. The non-compliance rate fell from 100% to 20% after a single pharmacist intervention. Significant improvement was observed in patients' mean blood pressures readings as well as the diabetic and lipid control. Positive impacts on healthcare resources utilization were also observed. / Chan Man Chi Grace. / "June 2005." / Adviser: Juliana C.N. Chan. / Source: Dissertation Abstracts International, Volume: 67-07, Section: B, page: 3730. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 126-151). / 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. / Abstract in English and Chinese. / School code: 1307.

I established the phage antibody library platform for the identification of specific antibodies. In the first part of my study, I tried to identify antibody against SARS-CoV. Two fragments on the spike protein, which is responsible for inducing viral entry, was chosen as target for the selection of antibody. An antibody was identified which can selectively recognize the SARS-CoV infected cells, but not non-infected cells. Although this antibody was found to retain no neutralizing ability, this specific antibody may have potential to develop for diagnostic purpose. / I utilized the phage system-based cloning method as an attractive approach to screen and identify virus-specific antibodies that can be encoded by the human genome. Once a useful phage clone is identified, unlimited amounts of human monoclonal virus-specific antibodies can be manufactured, and potentially applied clinically for prophylactic and therapeutic uses. The study focuses on two of these new infections, both of which cause severe respiratory disease: SARS and avian influenza. / Identification of specific antibodies, either for diagnostic or therapeutic use, was successfully demonstrated in the two infectious disease models. The phage antibody platform offers a fast and cost-effective method to identify phage antibodies, which can easily be converted to human viral specific monoclonal antibodies for clinical use. / In the 21st century, a number of novel infectious diseases emerged suddenly and spread rapidly, endangering the lives and well-being of people around the world. Severe acute respiratory syndrome (SARS) is a life threatening form of atypical pneumonia that ravaged Hong Kong, Taiwan, China, Canada and many cities in 2003. In the same year, novel avian influenza viruses infected human beings on two continents. Both of these diseases originated in animals and crossed over into the human population. These emerging diseases pose significant public health threats while providing a chilling reminder that another influenza pandemic could occur at any time. Thus, the development of effective therapeutics to control the disease is of paramount importance. Although several vaccines against SARS and avian influenza are available nowadays, the poor clinical performance and frequent mutation of viral strains may limit the practical use and value of the vaccines. Moreover, there are no promising antiviral drugs available for the treatment. Therefore, I aimed to develop an immunotherapy as an alternative treatment option against these diseases. / In the second part of my study, the extracellular domain of matrix protein of avian influenza virus was chosen as target for the selection of antibody. I successfully identified an antibody which can neutralize the avian influenza virus infection. This promising result indicated this antibody has potential to develop for therapeutic use and these antibodies can be easily manufactured in unlimited amounts for clinical application. / Leung, Ka Man. / Adviser: Kwok Pui Fung. / Source: Dissertation Abstracts International, Volume: 71-01, Section: B, page: 0212. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 112-123). / 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 Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese.

Asthma has long been considered as one of the most common health problems in the world. In Hong Kong, the prevalence of childhood asthma has increased from 4.8% in 1989, to 10.2% in 2002. In spite of the popularity of using Chinese herbs to treat asthma in Hong Kong, evidence on the effectiveness of herbal treatments is lacking. The Chinese herbal formula CUF2 is an innovative formula developed in the Institute of Chinese Medicine, The Chinese University of Hong Kong and it is composed of 5 commonly used Chinese herbs: Radix astragali, Cordyceps sinesis, Radix Scutellaria, Bulbus fritillariae cirrhosae and Radix stemonae. These herbs are chosen because of their well-known effects on either reducing coughing and sputum production, or anti-inflammatory and immunomodulatory activities. Based on the theoretical benefits of CUF2, we conducted a series of animal, in vitro and clinical studies to explore the efficacy, safety and mechanism of action of CUF2. / Following the establishment of the animal model, we have investigated the effect of CUF2 using this model of asthma. We found that 28 days pretreatment with CUF2 could reduce total cell number and eosinophilia in bronchoalveolar lavage fluid (BALF), prevent the eosinophil infiltration of airways, decrease pulmonary inflammatory cells, and reduce mucus and goblet cell hyperplasia. Especially in the reduction of goblet cell hyperplasia, we demonstrated that there was no significant difference between the effects of high dose CUF2 and dexamethasone (DEX). The eosinophilic immune-inflammatory responses in the airways in OA-sensitized/challenged rats were completely blocked by DEX returning to almost the same as those in normal rats, but the loss of thymus index and body weight were also observed. In contrast to the overall immunosuppressive effects of DEX, decreased production of inflammatory cytokines and chemokines [interleukin (IL)-4, tumor necrosis factor (TNF)-alpha, macrophage inflammatory protein (MIP)-2 and monocyte chemotactic protein-1 (MCP-1)], increased production of IL-10 and interferon-gamma (IFN-gamma) in BALF and no suppression of body weight and thymus index were demonstrated in the CUF2-treated groups. There was a dose-response relationship with more prominent effects seen with higher doses of CUF2. These findings indicate that the CUF2 has anti-airway inflammatory activity and exhibits immunomodulatory effect on Th1/Th2 responses in ovalbumin sensitized rats after allergen challenge, and this may imply its potential application to patients with allergic asthma. / In order to evaluate the clinical efficacy and safety, we conducted a multicenter, randomized, double blind, parallel and placebo controlled clinical trial. The same Chinese herbal formula was used in this clinical trial in 85 children aged 7-15 years with mild to moderate perennial asthma as an adjuvant therapy for 6 months. The primary outcome measure was the steroid dosage reduction. Other outcome measures included changes in disease severity score (DSS), lung function, serum concentrations of total IgE and the levels of some key allergy and inflammatory markers in peripheral blood, fractional exhaled nitric oxide (FENO), frequency of asthma attacks and quality of life (QOL). To assess safety, we did urinalysis, complete blood count, liver function and renal function at baseline and the end of the study. Drug compliance and adverse effects were also checked at each monthly visit. All patients were maintained on inhaled corticosteroid at their usual dose and dosing interval, and continued to receive short-acting, inhaled beta2-agonists as needed. There were no serious adverse events reported in the 6-month study period by any of the subjects. Hematological (except eosinophils count) and biochemical profiles (including renal function and liver function) remained within normal limits in the CUF2 group and placebo group at the end of the study. CUF2 was well tolerated in asthmatic children. Both CUF2 group and placebo group showed an improvement in most of clinical parameters. The dosage of inhaled corticosteroid was successfully reduced in both groups. Both groups had similar decrease in DSS, improved QOL and improved lung function parameter PEFR (L/min). Although these parameters showed no statistically significant difference between two groups, the percentage of eosinophils and lymphocytes were significantly decreased in CUF2 group as compared with the placebo group. The CUF2 group also showed improved diary symptom score, reduced expression of TNF-alpha and slight increase in anti-inflammation cytokine IL-18 in the blood. A trend of greater improvement in frequency of upper respiratory infection (URI) in CUF2 group was noted, but no statistical significance was attained. The changes in lung function parameter FEV1%, FENO, frequency of asthma attacks and serum concentrations of total IgE, IgE HDM, IgE cat, cockroach, TARC, LTB4 and LTC4D4E4 showed no statistically significant difference CUF2 group and placebo group. Overall, our data demonstrated that CUF2 treatment had some immunomodulatory effect in childhood asthma. Our findings should support further investigations of Chinese herbal medicine in the area of asthma without steroid therapy. / In the animal study, firstly we attempted to establish a novel murine model of asthma. We adopted a modified sensitization procedure using 10-point subcutaneous and intraperitoneal injections of Ovalbumin (OA) with freshly prepared Al(OH)3 and successfully induced severe airway allergic reactions in young Sprague Dawley (SD) rats. In this SD rat model, allergen exposure triggered accumulation of inflammation cells and eosinophils in the airway submucosa and goblet cells hyperplasia in mucosa, lung function test revealed obstructive lung function changes that included increase of lung resistance (RL) and decrease of dynamic lung compliance (Cdyn). Cytokine and chemokine assays showed that there was a change of the TH1/Th2 balance as illustrated by the high Th2 (interleukin-4)/Th1 (interferon-gamma) ratio. These results demonstrated the feasibility and validity of the SD rat model for studying allergic asthma. This SD model is much cheaper and readily available than the Brown Norway rat model and may facilitate further drug trial in asthma. / In the in vitro study, we investigated the effect of CUF2 on the release of cytokines and/or gene expression using human mast cell line HMC-1, human bronchial epithelial cell line BEAS-2B, peripheral blood mononuclear cells (PBMCs) from healthy subjects and airway cells present in induced sputum from asthmatic patients. We have shown (1) the CUF2 had no cytotoxic effects in final working concentration; (2) CUF2 had inhibitory effects on IL-6, TNF-alpha and granulocyte-macrophage colony-stimulating factor (GM-CSF) secretion from HMC-1 in a dose-dependent manner. However, no reduction of IL-8 production in HMC-1 was demonstrated. (3) In addition, study of the effect of CUF2 on the expression of cytokine gene from HMC-1 showed that IL-4, IL-6 and GM-CSF mRNA expressions were down regulated at 24 hours, 24 hours, 16 hours and 24 hours of time points, respectively. No effects on IL-8 and TNF-alpha mRNA expression was observed. (4) Furthermore, CUF2 also significantly inhibited in vitro IL-6 and GM-CSF secretion in TNF-alpha stimulated BEAS-2B cell and reduced GM-CSF production in airway cells present in induced sputum from asthmatic children. (5) We observed that CUF2 enhanced TNF-alpha and IL-6 production but did not alter the levels of GM-CSF and IL-8 in mitogen-stimulated PBMCs from health subject. These findings suggest that pharmacological activities of the CUF2 may be mediated by regulating the production of cytokines in human mast cell, bronchial epithelial cell, airway cell and PBMCs. / In this study, a novel animal asthma model has been established. This model has extensively characterized and exhibited several inflammatory, immunological features that resemble those of human asthma and may facilitate further drug trial in asthma. CUF2 showed its efficacy treating the animal model of allergic asthma. In vitro study also provided evidence of its beneficial dichotomous effects on cytokine and chemokine production in HMC-1, PBMCs and airway cells. A multi-center, randomized, double blind, placebo controlled clinical trial showed that CUF2 had a certain degree of clinical efficacy. Furthermore, the use of CUF2, with the study dose and treatment period, was safe. The efficacy of individual ingredient and the mechanism of CUF2 have not been clarified and further investigations are warranted. In conclusion, our results provided evidence of the potential beneficial effect of CUF2 on immune system functions and supported the potential use of TCM as therapeutic drugs for allergic inflammatory diseases. / by Wong Yeuk Oi. / "September 2005." / Advisers: Yn Tz Sung; Kowk Pui Fung. / Source: Dissertation Abstracts International, Volume: 67-11, Section: B, page: 6296. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 330-349). / 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.

Leung, Jackie. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 117-131). / Abstract also in Chinese. / Abstract --- p.i / 論文摘要 --- p.iii / Acknowledgement --- p.iv / List of publications --- p.vi / List of Tables --- p.vi / List of Figures --- p.vi / Table of Contents --- p.ix / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1. --- Liver cancer --- p.1 / Chapter 1.1.1. --- Hepatocellular Carcinoma (HCC) --- p.2 / Chapter 1.2. --- Lung Cancer --- p.5 / Chapter 1.3. --- Pteris semipinnata L --- p.8 / Chapter 1.4. --- Extract of PsL: 5F --- p.10 / Chapter 1.5. --- Animal models in chemotherapy researches --- p.13 / Chapter 1.5.1. --- Model of HCC --- p.13 / Chapter 1.5.2. --- Model of lung cancer --- p.15 / Chapter 1.6. --- Apoptosis: Significance of programmed cell death --- p.17 / Chapter 1.6.1. --- The extrinsic pathway --- p.18 / Chapter 1.6.2. --- The intrinsic pathway --- p.19 / Chapter 1.7. --- Apoptotic molecules related to this study --- p.22 / Chapter 1.7.1. --- Bcl-2 family --- p.22 / Chapter 1.7.1.1. --- Bax --- p.22 / Chapter 1.7.1.2. --- Bcl-2 --- p.23 / Chapter 1.7.2. --- Nuclear factor kappa B --- p.25 / Chapter 1.7.3. --- Inducible nitric oxide synthase --- p.27 / Chapter 1.8. --- Side-effects of chemotherapy --- p.29 / Chapter 1.8.1. --- Chemotherapy and liver dysfunction --- p.30 / Chapter 1.8.2. --- Nephrotoxicity of chemotherapeutic agents --- p.31 / Chapter 1.9. --- Aim of study --- p.33 / Chapter Chapter 2: --- Materials and Methodology --- p.34 / Chapter 2.1. --- Animals --- p.34 / Chapter 2.1.1. --- HCC model --- p.34 / Chapter 2.1.2. --- Lung cancer model --- p.35 / Chapter 2.2. --- Tumors induction --- p.36 / Chapter 2.2.1. --- HCC induction in C3H/HeJ mice --- p.36 / Chapter 2.2.2. --- Lung cancer induction in A/J mice --- p.37 / Chapter 2.3. --- 5F preparation --- p.38 / Chapter 2.4. --- 5F treatment --- p.39 / Chapter 2.5. --- Harvest of samples and tissues --- p.41 / Chapter 2.6. --- Tumor assessment --- p.43 / Chapter 2.7. --- Investigation of apoptosis and cell proliferation --- p.44 / Chapter 2.8. --- Immunohistochemistry --- p.47 / Chapter 2.9. --- Biochemical test --- p.51 / Chapter 2.9.1. --- Liver Function Tests (LFT) --- p.52 / Chapter 2.9.1.1. --- Aspartate aminotransferase (AST) & Alanine aminotransferase (ALT) --- p.52 / Chapter 2.9.2. --- Renal Function Test (RFT) --- p.53 / Chapter 2.9.2.1. --- Serum creatinine level (CRE) --- p.53 / Chapter 2.9.2.2. --- Blood Urea Nitrogen index (BUN) --- p.54 / Chapter 2.10. --- Statistical analysis --- p.55 / Chapter Chapter 3: --- Results --- p.56 / Chapter 3.1. --- Anti-tumor effect of 5F is dose- dependent --- p.56 / Chapter 3.2. --- 5F reduces cell proliferation and induces apoptosis in-vivo --- p.60 / Chapter 3.3. --- Effects of 5F on apoptotic signaling molecules --- p.68 / Chapter 3.3.1. --- 5F up-regulates pro-apoptotic Bax and Bak --- p.68 / Chapter 3.3.2. --- 5F down-regulates anti-apoptotic NF-kappa B and Bcl-2 --- p.76 / Chapter 3.3.3. --- 5F up-regulated iNOS in HCC but not in lung cancer --- p.88 / Chapter 3.3.4. --- Regulation on Erk1/2 was associated with treatment of 5F --- p.93 / Chapter 3.4. --- Side-effect studies of 5F --- p.97 / Chapter Chapter 4: --- Discussion --- p.105 / Chapter Chapter 5: --- Conclusion --- p.116 / Bibliography --- p.117

Chan, Man Hung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 111-126). / Abstract also in Chinese. / Examination Committee List --- p.ii / Declaration --- p.iii / Acknowledgements --- p.iv / 摘要（Abstract in Chinese) --- p.vi / Abstract --- p.viii / List of Abbreviations --- p.x / List of Figures and Tables --- p.xii / Table of Content --- p.xiv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Background of photodynamic therapy (PDT) --- p.2 / Chapter 1.1.1 --- History of PDT --- p.2 / Chapter 1.1.2 --- Photochemistry --- p.3 / Chapter 1.1.3 --- Principal stages of PDT --- p.5 / Chapter 1.1.4 --- Light sources of PDT --- p.6 / Chapter 1.2 --- Anti-tumor effect of PDT --- p.8 / Chapter 1.2.1 --- Mode of cell death --- p.8 / Chapter 1.2.2 --- PDT-induced anti-tumor immunity --- p.9 / Chapter 1.3 --- Clinical applications of PDT --- p.11 / Chapter 1.3.1 --- Photofrin® --- p.11 / Chapter 1.3.2 --- Clinical applications of PDT --- p.13 / Chapter 1.3.3 --- Challenges of PDT for clinical applications --- p.15 / Chapter 1.4 --- The development of new photosensitizers --- p.16 / Chapter 1.4.1 --- Targeted PDT --- p.16 / Chapter 1.4.2 --- Phthalocyanine --- p.18 / Chapter 1.5 --- Objective of my study --- p.21 / Chapter Chapter 2 --- Materials and Methods --- p.23 / Chapter 2.1 --- Synthesis of glucosylated silicon(IV) phthalocyanine (SiPcGlu) --- p.24 / Chapter 2.2 --- In vitro studies --- p.24 / Chapter 2.2.1 --- Cell line and culture conditions --- p.24 / Chapter 2.2.2 --- Photodynamic treatment --- p.25 / Chapter 2.2.3 --- Cell viability assay --- p.27 / Chapter 2.2.4 --- Light dose effect on the photocytotoxicity of SiPcGlu-PDT --- p.27 / Chapter 2.2.5 --- Determination of reactive oxygen species (ROS) production by SiPcGlu-PDT --- p.29 / Chapter 2.2.6 --- Effect of antioxidants on the photocytotoxicity of SiPcGlu-PDT --- p.29 / Chapter 2.2.7 --- Determination of ROS production after SiPcGlu-PDT --- p.30 / Chapter 2.2.8 --- Glucose competitive assay --- p.30 / Chapter 2.2.9 --- Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay --- p.30 / Chapter 2.2.10 --- DNA fragmentation analysis by gel electrophoresis --- p.31 / Chapter 2.2.11 --- Annexin-V & propidium iodide staining assay --- p.32 / Chapter 2.2.12 --- Subcellular localization studies --- p.33 / Chapter 2.2.13 --- Detection of mitochondrial superoxide production --- p.34 / Chapter 2.2.14 --- Assessment of mitochondrial membrane potential --- p.34 / Chapter 2.2.15 --- Caspase-3 activity assay --- p.35 / Chapter 2.2.16 --- "Western blot analyses for cytochrome c, caspase-3, PARP and glucose-regulated protein 78 (GRP78)" --- p.36 / Chapter 2.2.17 --- Ca2+ release from endoplasmic reticulum (ER) --- p.37 / Chapter 2.3 --- In vivo studies --- p.37 / Chapter 2.3.1 --- HT29 tumor-bearing nude mice model --- p.37 / Chapter 2.3.2 --- In vivo photodynamic treatment --- p.39 / Chapter 2.3.3 --- Biodistribution of SiPcGlu --- p.39 / Chapter 2.3.4 --- Assay for plasma enzyme activities --- p.40 / Chapter 2.4 --- Statistical analysis --- p.41 / Chapter Chapter 3 --- Results --- p.42 / Chapter 3.1 --- In vitro studies --- p.43 / Chapter 3.1.1 --- SiPcGlu-PDT induced cytotoxicity on HT29 cells --- p.43 / Chapter 3.1.2 --- Light dose effect on cytotoxicity by SiPcGlu-PDT --- p.46 / Chapter 3.1.3 --- SiPcGlu-PDT induced ROS production --- p.48 / Chapter 3.1.4 --- SiPcGlu-PDT induced cell death through Type I and II photoreactions --- p.48 / Chapter 3.1.5 --- ROS production after SiPcGlu-PDT --- p.51 / Chapter 3.1.6 --- Glucose competitive Assay --- p.55 / Chapter 3.1.7 --- SiPcGlu-PDT induced apoptosis in HT29 cells --- p.57 / Chapter 3.1.8 --- Subcellular localization of SiPcGlu --- p.61 / Chapter 3.1.9 --- SiPcGlu-PDT induced mitochondrial changes --- p.66 / Chapter 3.1.10 --- SiPcGlu-PDT induced caspase activation --- p.68 / Chapter 3.1.11 --- SiPcGlu-PDT increased expression of ER chaperone GRP78 --- p.72 / Chapter 3.1.12 --- SiPcGlu-PDT induced release of Ca2+ from ER --- p.72 / Chapter 3.2 --- In vivo studies --- p.75 / Chapter 3.2.1 --- In vivo photodynamic activities --- p.75 / Chapter 3.2.2 --- Tissue distribution of SiPcGlu --- p.77 / Chapter 3.2.3 --- Analysis of intrinsic toxicity --- p.77 / Chapter Chapter 4 --- Discussion --- p.80 / Chapter 4.1 --- Physical Properties of SiPcGlu --- p.81 / Chapter 4.2 --- In vitro studies --- p.82 / Chapter 4.2.1 --- SiPcGlu-PDT exhibits a high potency in killing HT29 cells --- p.82 / Chapter 4.2.2 --- ROS production is responsible for the cytotoxic effect of SiPcGlu-PDT --- p.83 / Chapter 4.2.3 --- SiPcGlu-PDT induced apoptosis in HT29 cells --- p.85 / Chapter 4.2.4 --- SiPcGlu is localized in various membranous organelles --- p.87 / Chapter 4.2.5 --- SiPcGlu-PDT induced mitochondria-mediated apoptosis --- p.89 / Chapter 4.2.6 --- SiPcGlu-PDT induced ER stress --- p.93 / Chapter 4.3 --- In vivo studies --- p.96 / Chapter 4.3.1 --- SiPcGlu failed to target to tumor tissues --- p.96 / Chapter 4.3.2 --- SiPcGlu-PDT induced retardation in tumor growth --- p.99 / Chapter 4.3.3 --- SiPcGlu is a safe photosensitizer for PDT --- p.101 / Chapter Chapter 5 --- Conclusion and Future Perspectives --- p.103 / Chapter 5.1 --- Conclusion --- p.104 / Chapter 5.2 --- Future Perspectives --- p.106 / Chapter 5.2.1 --- In vitro studies --- p.106 / Chapter 5.2.1.1 --- Lysosomal pathway to cell death --- p.106 / Chapter 5.2.2 --- In vivo studies --- p.107 / Chapter 5.2.2.1 --- Pharmacokinetic studies --- p.107 / Chapter 5.2.2.2 --- Eradication of HT29 tumor by repeated dose of SiPcGlu --- p.108 / Chapter 5.2.2.3 --- SiPcGlu-PDT-induced anti-tumor immunity --- p.108 / Chapter 5.2.2.4 --- Enhancement of tumor selectivity by conjugating with biomolecules --- p.109 / References --- p.110

Thesis (M.S.)--University of South Florida, 2009. / Title from PDF of title page. Document formatted into pages; contains 35 pages. Includes bibliographical references.

Background. Both basal-like subtype and BRCA1-related breast cancers tend to have a poor overall prognosis and lack of effective treatments. Given that the lung cancer drug gefitinib and the leukemia drug dasatinib inhibit proteins also belonging to the molecular signature of this subtype, we and others hypothesized that they might be useful therapies for those two breast cancer subgroups. / Methods. Eight breast cancer cell lines were characterized by immunohistochemistry and western blotting and were treated with both drugs. Response was measured by using the sulphorhodamine B (SRB) assay. / Results. Two out of six basal-like cell lines were sensitive to gefitinib and five of six to dasatinib. BRCA1-related breast cancers were also responsive to dasatinib (three out of four). Moreover, EGFR and caveolin-1 act as markers for dasatinib sensitivity, but do not appear to be the primary targets of this drug. The presence of SRC but not ABL is necessary to achieve a response to dasatinib. / Conclusion. Dasatinib is more effective in the treatment of basal-like breast cancers than gefitinib and acts by inhibiting SRC and other molecules that are yet to be determined.

Dissertation (Ph.D.)--University of South Florida, 2007. / Title from PDF of title page. Document formatted into pages; contains 192 pages. Includes vita. Includes bibliographical references.