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
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324376 |
| Date | January 2003 |
| Contributors | Tang, Yuk Kwan., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xvii, 144 leaves : ill. (some col.) ; 30 cm. |
| Rights | Use 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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