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Suppressive effect on the secretion of hepatitis B surface antigen (HBsAg) by Phyllanthus urinaria in alexander cells.

Tang Yuk Kwan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 130-144). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.iv / List of Abbreviations --- p.v / List of Figures --- p.ix / List of Tables --- p.xii / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Hepatitis B Virus (HBV) --- p.1 / Chapter 1.1.1 --- Epidemiology --- p.1 / Chapter 1.1.2 --- History of HBV --- p.2 / Chapter 1.1.3 --- Hepatitis B in Hong Kong --- p.3 / Chapter 1.2 --- Virology --- p.4 / Chapter 1.2.1 --- Hepadnavirus Family --- p.4 / Chapter 1.2.2 --- HBV Particles Types --- p.4 / Chapter 1.2.3 --- HBV Genome --- p.6 / Chapter 1.2.4 --- HBV Life Cycle --- p.8 / Chapter 1.2.5 --- Hepatitis B Surface Antigen --- p.14 / Chapter 1.3 --- HBV Immunobiology During Viral Infection --- p.17 / Chapter 1.3.1 --- Acute Hepatitis B Virus Infection --- p.19 / Chapter 1.3.2 --- Chronic Hepatitis B Virus Infection --- p.23 / Chapter 1.3.3 --- Cytokines in Suppression of HBV Infection --- p.23 / Chapter 1.3.4 --- The mechanism of HBV Persistence --- p.26 / Chapter 1.4 --- HBV Therapy --- p.28 / Chapter 1.4.1 --- Interferon alpha (IFN-α) --- p.29 / Chapter 1.4.2 --- Lamivudine --- p.31 / Chapter 1.5 --- Phyllanthus --- p.32 / Chapter 1.5.1 --- In vitro study --- p.32 / Chapter 1.5.2 --- In vivo study --- p.33 / Chapter 1.5.3 --- Clinical Trials --- p.34 / Chapter 1.5.4 --- Anti-tumor Effect of P. amarus --- p.35 / Chapter 1.5.5 --- Active component(s) of P. am arus --- p.35 / Chapter 1.6 --- Alexander cells --- p.37 / Chapter 1.7 --- Objectives --- p.38 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials --- p.39 / Chapter 2.1.1 --- Phyllanthus urinaria --- p.39 / Chapter 2.2 --- In vitro study --- p.39 / Chapter 2.2.1 --- Materials --- p.39 / Chapter 2.2.1.1 --- Cell Lines --- p.39 / Chapter 2.2.2 --- Reagents and Buffers --- p.40 / Chapter 2.2.2.1 --- Phosphate Buffered Saline (PBS) --- p.40 / Chapter 2.2.2.2 --- Reagents and Buffers for mRNA Genes Expression Analyses --- p.40 / Chapter 2.2.2.3 --- Reagents and Buffers for Protein Analyses --- p.41 / Chapter 2.2.2.4 --- Reagents for Purification of Splenocytes --- p.43 / Chapter 2.2.3 --- Methods --- p.43 / Chapter 2.2.3.1 --- MTT Assay --- p.43 / Chapter 2.2.3.2 --- IMX Assay --- p.43 / Chapter 2.2.3.3 --- Semi-quantitative RT-PCR --- p.44 / Chapter 2.2.3.3.1 --- Extraction of RNA --- p.44 / Chapter 2.2.3.3.2 --- Reverse Transcription Polymerase Chain Reaction (RT-PCR) --- p.45 / Chapter 2.2.3.3.3 --- Polymerase Chain Reaction (PCR) --- p.46 / Chapter 2.2.3.4 --- Protein Analysis --- p.48 / Chapter 2.2.3.4.1 --- Extraction of Protein --- p.48 / Chapter 2.2.3.4.2 --- Determination of Protein Concentrations --- p.48 / Chapter 2.2.3.4.3 --- Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.49 / Chapter 2.2.3.4.4 --- Electroblotting of Protein --- p.50 / Chapter 2.2.3.4.5 --- Immunoblotting of Protein --- p.50 / Chapter 2.2.3.4.6 --- Enhanced Chemiluminescence (ECL) Assay --- p.51 / Chapter 2.2.3.5 --- Assay of preSl Promoter Activity --- p.51 / Chapter 2.2.3.5.1 --- Cloning of preSl Promoter from HBV Genome --- p.51 / Chapter 2.2.3.5.2 --- Transfection --- p.52 / Chapter 2.2.3.5.3 --- Luciferase Assay --- p.53 / Chapter 2.2.3.6 --- Pilot Experiments for the Investigation of Immunostimulatory Effect of (aq) P. urinaria --- p.53 / Chapter 2.2.3.6.1 --- Purification of Splenocytes --- p.53 / Chapter 2.2.3.6.2 --- Cytotoxicity Assay of Splenocytes --- p.54 / Chapter 2.2.3.6.3 --- [3Hl-thymidine Uptake Assay --- p.54 / Chapter 2.2.3.6.4 --- Purification of Macrophages --- p.55 / Chapter 2.3 --- In vivo Assay --- p.56 / Chapter 2.3.1 --- Materials --- p.56 / Chapter 2.3.1.1 --- Balb/c Mice --- p.56 / Chapter 2.3.1.2 --- Reagents for Cytokine Assay --- p.56 / Chapter 2.3.1.3 --- Reagents for Flow Cytometric Analysis of T Cell Subpopulations --- p.57 / Chapter 2.3.2 --- Methods --- p.57 / Chapter 2.3.2.1 --- Purification of Splenocytes --- p.57 / Chapter 2.3.2.2 --- Enzyme Assay --- p.57 / Chapter 2.3.2.3 --- Flow Cytometric Analysis of T Cell Subpopulations --- p.59 / Chapter 2.3.2.4 --- Cytokine Assays --- p.60 / Chapter 2.4 --- Statistical Analysis --- p.63 / Chapter Chapter 3 --- Results / Chapter 3.1 --- In vitro Assays --- p.64 / Chapter 3.1.1 --- Study of the Cytotoxicity of Crude Extract of (aq) P. urinaria on Alexander Cells by the MTT Assay --- p.64 / Chapter 3.1.2 --- Study of the Cytotoxicity of Crude Extract of (aq) P. urinaria on WRL 68 Cells by the MTT Assay --- p.66 / Chapter 3.1.3 --- Study of the Cytotoxicity of Crude Extract of (aq) P. urinaria on Primary Mouse Macrophages by the MTT Assay --- p.67 / Chapter 3.1.4 --- Effect on HBsAg Secretion by the IMX Assay --- p.69 / Chapter 3.1.5 --- Effect of Viral Gene Expression by Semi-quantitative RT-PCR --- p.73 / Chapter 3.1.6 --- Effect of Intracellular Viral Proteins Expression by Western Blotting Analyses --- p.75 / Chapter 3.1.7 --- preSl Promoter Activity in Alexander Cell Line and Hep3B Cell Line --- p.77 / Chapter 3.1.8 --- Effect of (aq) P. urinaria on preSl Promoter Activity in Hep3B Cell Line --- p.80 / Chapter 3.1.9 --- Pilot Experiments for the Investigation of Immunomodulatory Effect of (aq) P. urinaria --- p.82 / Chapter 3.1.10 --- Study of the Cytotoxicity of Crude Extract of (aq) P. urinaria on Primary Splenocytes by the MTT Assay --- p.85 / Chapter 3.2 --- In vivo Assay --- p.88 / Chapter 3.2.1 --- Study of the Immunomodulatory Effect of the Crude Extract of (aq) P. urinaria on Normal Balb/c Mice by [3H]-thymidine Uptake Assay --- p.88 / Chapter 3.2.2 --- Study of the Toxicity of Crude Extract of (aq) P. urinaria on Heart and Liver of Normal Balb/c Mice by Enzyme Assay --- p.91 / Chapter 3.2.3 --- Flow Cytometric Analysis of T Cell Subpopulations --- p.94 / Chapter 3.2.4 --- Analysis of Stimulatory Effect on Four Different Cytokines by ELISA --- p.100 / Chapter Chapter 4 --- Discussion / Chapter 4.1 --- In vitro Assay --- p.110 / Chapter 4.2 --- In vivo Assay --- p.116 / Chapter 4.3 --- Conclusions --- p.127 / Chapter 4.4 --- Future Prospects --- p.129 / References --- p.130

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324376
Date January 2003
ContributorsTang, Yuk Kwan., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xvii, 144 leaves : ill. (some col.) ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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