Wang Ying. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 124-135). / Abstracts in English and Chinese. / Acknowledgements / Abstract / 摘要 / Table of contents / List of figures / List of tables / List of abbreviations / CHAPTER / Chapter 1. --- Introduction / Chapter 1.1 --- Background of SARS and epidemiology / Chapter 1.2 --- SARS symptoms and infected regions / Chapter 1.3 --- SARS virus / Chapter 1.4 --- Treatment for SARS at present / Chapter 1.5 --- Vaccine development is a more effective way to fight against SARS / Chapter 1.6 --- Vaccine candidates / Chapter 1.6.1 --- Truncated S protein as a vaccine candidate / Chapter 1.6.2 --- Full-length N protein as a vaccine candidate / Chapter 1.7 --- E.coli expression system / Chapter 1.8 --- Baculovirus expression system / Chapter 1.8.1 --- Characteristics of baculovirus / Chapter 1.8.2 --- Infection cycle of baculovirus / Chapter 1.8.3 --- Control of viral gene expression in virus-infected cells / Chapter 1.8.4 --- Merits of baculovirus expression system / Chapter 1.9 --- Aim of study / Chapter 2. --- "Bacterial expression and purification of rS1-1000(E), rS401-1000(E) and rN(E)" / Chapter 2.1 --- Introduction / Chapter 2.2 --- Materials / Chapter 2.2.1 --- Reagents for bacterial culture / Chapter 2.2.2 --- Reagents for agarose gel electrophoresis / Chapter 2.2.3 --- 2'-deoxyribonucleoside 5'-triphosphate (dNTP) mix for polymerase chain reaction (PCR) / Chapter 2.2.4 --- Sonication buffer / Chapter 2.2.5 --- Reagents for immobilized metal affinity chromatography (IMAC) purification / Chapter 2.2.6 --- Reagents for gel filtration chromatography / Chapter 2.2.7 --- Reagents for sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) / Chapter 2.2.8 --- Reagents for Western blotting / Chapter 2.3 --- Methods / Chapter 2.3.1 --- General techniques in molecular cloning / Chapter 2.3.2 --- "PCR amplification of the S1-400,S401-1000" / Chapter 2.3.3 --- Construction of clone pET-S 1-400 and PET-s401-1000 / Chapter 2.3.4 --- Construction of clone pAC-N / Chapter 2.3.5 --- Expression / Chapter 2.3.6 --- Inclusion bodies preparation / Chapter 2.3.7 --- Inclusion bodies solubilization using urea / Chapter 2.3.8 --- Protein refolding by rapid dilution and dialysis / Chapter 2.3.9 --- Purification of recombinant protein by nickel ion chelating Sepharose fast flow column (IMAC) / Chapter 2.3.10 --- Gel filtration chromatography for further purification / Chapter 2.3.11 --- Bradford assay for the protein concentration analysis / Chapter 2.3.12 --- Protein analysis / Chapter 2.4 --- Results / Chapter 2.4.1 --- SDS-PAGE analysis of the expressed proteins / Chapter 2.4.2 --- Western blot analysis of the bacterial cell lysate / Chapter 2.4.3 --- Protein purification by IMAC / Chapter 2.4.4 --- Purification of rS401-1000(E) by gel filtration / Chapter 2.4.5 --- Determination of production yield of recombinant fusion proteins / Chapter 2.5 --- Discussion / Chapter 2.5.1 --- Expression vector selected for rS1-400(E) and rS401-1000(E) expression / Chapter 2.5.2 --- Protein expression in E.coli / Chapter 2.5.3 --- Purification process / Chapter 3. --- Baculovirus expression and purification of rS401-1000(ACN) and rN(BMN) protein / Chapter 3.1 --- Introduction / Chapter 3.2 --- Materials / Chapter 3.2.1 --- Reagents for insect cell culture and virus work / Chapter 3.3 --- Methods / Chapter 3.3.1 --- "PCR amplification of N and cloning of S401-1000, N genes into the transfer vector pVL1393" / Chapter 3.3.2 --- Cloning of S401-1000 into transfer vector pFastBac HT B / Chapter 3.3.3 --- Virus works / Chapter 3.3.4 --- Identification of recombinant BmNPV or AcMNPV / Chapter 3.3.5 --- Manipulation of silkworm / Chapter 3.3.6 --- Mouse immunization for polyclonal antibody against rN(E) protein / Chapter 3.4 --- Results / Chapter 3.4.1 --- Expression of rN(BMN) in baculovirus / Chapter 3.4.2 --- Expression of rS401-1000(BMN) and rS401-1000(ACN) in baculovirus / Chapter 3.5 --- Discussion / Chapter 3.5.1 --- The expression level of rN(BMN) in both in vitro and invivo / Chapter 3.5.2 --- The rS401-1000(ACN) protein expression level in vitro / Chapter 3.5.3 --- Failure in generating rS401-1000(BMN) / Chapter 3.5.4 --- Purification process of rN(BMN) by IMAC / Chapter 4. --- "Characterization of recombinant rS1-400(E), rN(E), rN(BMN), rS401_1000(E) and rS401-1000(ACN)" / Chapter 4.1 --- Introduction / Chapter 4.2 --- Materials / Chapter 4.2.1 --- Reagents for enzyme-linked immunosorbent assay (ELISA) / Chapter 4.2.2 --- Reagents for purification of human IgG / Chapter 4.2.3 --- Source and identity of Immune sera / Chapter 4.3 --- Methods / Chapter 4.3.1 --- ELISA / Chapter 4.3.2 --- Purification process of human IgG / Chapter 4.4 --- Results / Chapter 4.4.1 --- Validation of Immune sera using SARS viral lysate / Chapter 4.4.2 --- Immunoreactivities of rS1-400(E) and rN(E) against pooled patients sera and normal human serum / Chapter 4.4.3 --- Immunoreactivity comparison of rN(E) and rN(BMN) / Chapter 4.4.4 --- Comparison of the immunoreactivities of rS401-1000(E) and rS401-1000(ACN) / Chapter 4.4.5 --- Immunoreactivity of SARS related proteins against Anti-SARS Antibody (Equine) / Chapter 4.5 --- Discussion / Chapter 4.5.1 --- Comparison of the immunoreactivities of SARS related proteins expressed in the present study / References

Parkash Amarender. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 158-172). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Table of contents --- p.ii / Abstract --- p.xi / 撮要 --- p.xiv / List of Abbreviations --- p.xvi / List of Tables --- p.xvii / List of Figures --- p.xix / Chapter CHAPTER 1. --- INTRODUCTION / Chapter 1.1 --- Ribosome-inactivating proteins (RIPs) --- p.3 / Chapter 1.2 --- General Properties of RIPs --- p.5 / Chapter 1.2.1 --- Structure --- p.5 / Chapter 1.2.1.1 --- Type I and Type II RIPs --- p.5 / Chapter 1.2.1.2 --- Small RIPs --- p.10 / Chapter 1.2.2 --- Distribution --- p.12 / Chapter 1.2.3 --- Physicochemical properties --- p.15 / Chapter 1.3 --- Enzymatic activities of RIPs --- p.17 / Chapter 1.3.1 --- N-glycosidase activity --- p.17 / Chapter 1.3.2 --- Polynucleotide:adenosine glycosidase activity --- p.21 / Chapter 1.3.3 --- Ribonuclease (RNase) activity --- p.24 / Chapter 1.3.4 --- Deoxyribonucleolytic (DNase) activity --- p.25 / Chapter 1.3.5 --- Multiple depurination --- p.26 / Chapter 1.3.6 --- Inhibition of protein synthesis --- p.27 / Chapter 1.4 --- Biological activities of RIPs --- p.29 / Chapter 1.4.1 --- Interaction of ribosome-inactivating proteins with cells --- p.29 / Chapter 1.4.1.1 --- Internalization of type 1 ribosome-inactivating proteins --- p.29 / Chapter 1.4.1.2 --- Internalization of type 2 ribosome-inactivating proteins --- p.32 / Chapter 1.4.2 --- Effects on laboratory animals --- p.33 / Chapter 1.4.3 --- Immunosuppressive activity --- p.33 / Chapter 1.4.4 --- Abortifacient activity --- p.34 / Chapter 1.4.5 --- Antiviral activity --- p.35 / Chapter 1.5 --- Physiological roles of RIPs --- p.37 / Chapter 1.6 --- Applications of RIPs --- p.39 / Chapter 1.6.1 --- Possible uses in experimental and clinical medicine --- p.39 / Chapter 1.6.1.1 --- Anti-tumor therapy --- p.40 / Chapter 1.6.1.2 --- Immune disorders --- p.42 / Chapter 1.6.1.3 --- Neuroscience research --- p.43 / Chapter 1.6.2 --- Applications in agriculture --- p.44 / Chapter 1.7 --- Arginine/Glutamate Rich Polypeptides (AGRPs) --- p.46 / Chapter 1.8 --- Objectives of the present study --- p.48 / Chapter 1.8.1 --- Rationale of the study --- p.48 / Chapter 1.8.2 --- Outline of the thesis --- p.50 / Chapter Chapter 2 --- Materials and methods / Chapter 2.1 --- Introduction --- p.52 / Chapter 2.2 --- Materials and methods --- p.54 / Chapter 2.2.1 --- Materials --- p.54 / Chapter 2.2.2 --- Preparation of crude extract --- p.55 / Chapter 2.2.3 --- Purification of proteins --- p.55 / Chapter 2.2.4 --- Molecular weight determination with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) --- p.61 / Chapter 2.2.5 --- Protein determination --- p.64 / Chapter 2.2.6 --- N-terminal amino acid sequence --- p.64 / Chapter 2.2.7 --- Preparation of rabbit reticulocyte lysate --- p.65 / Chapter 2.2.8 --- Assay for cell-free protein synthesis- inhibiting activity --- p.65 / Chapter 2.2.9 --- Assay for N-glycosidase activity --- p.66 / Chapter 2.2.10 --- Assay for ribonuclease activity --- p.70 / Chapter 2.2.11 --- Assay for antifungal activity --- p.71 / Chapter 2.2.12 --- Assay for dehydrogenase activity --- p.71 / Chapter Chapter 3 --- Purification and characterization of proteins from their respective sources. / Chapter 3.1. --- Purification and Characterization of Hispidin from Hairy melon (Benincasa hispida var. chieh-qua) / Chapter 3.1.1. --- Introduction --- p.73 / Chapter 3.1.2. --- Results --- p.76 / Chapter 3.1.2.1. --- Purification --- p.78 / Chapter 3.1.2.2. --- Molecular weight determination --- p.84 / Chapter 3.1.2.3. --- N-terminal amino acid sequence --- p.85 / Chapter 3.1.2.4. --- Assay for cell-free protein synthesis-inhibiting activity --- p.86 / Chapter 3.1.2.5. --- Assay for N-glycosidase activity --- p.87 / Chapter 3.1.2.6. --- Assay for ribonuclease activity --- p.88 / Chapter 3.1.2.7. --- Assay for dihydrodiol dehydrogenase activity --- p.88 / Chapter 3.1.2.8. --- Assay for antifungal activity --- p.89 / Chapter 3.1.2.9. --- "Assessment of purity, yield and activity" --- p.91 / Chapter 3.1.3. --- Discussion --- p.92 / Chapter 3.2. --- Purification and Characterization of Momorchin from Dried Bitter Gourd (Momordica charantia) Seeds / Chapter 3.2.1. --- Introduction --- p.95 / Chapter 3.2.2. --- Results --- p.99 / Chapter 3.2.2.1. --- Purification --- p.100 / Chapter 3.2.2.2. --- Molecular weight determination --- p.103 / Chapter 3.2.2.3. --- N-terminal amino acid sequence --- p.104 / Chapter 3.2.2.4. --- Assay for cell-free protein synthesis- inhibiting activity --- p.105 / Chapter 3.2.2.5. --- Assay for ribonuclease activity --- p.105 / Chapter 3.2.2.6. --- Assay for N-glycosidase activity --- p.106 / Chapter 3.2.2.7. --- "Assessment of purity, yield and activity" --- p.107 / Chapter 3.2.3. --- Discussion --- p.108 / Chapter 3.3.3. --- Purification and Characterization of Luffacylin from Sponge Gourd (Luffa cylindrica) / Chapter 3.3.1. --- Introduction --- p.110 / Chapter 3.3.2. --- Results --- p.113 / Chapter 3.3.2.1. --- Purification --- p.115 / Chapter 3.3.2.2. --- Molecular weight determination --- p.119 / Chapter 3.3.2.3. --- N-terminal amino acid sequencing --- p.120 / Chapter 3.3.2.4. --- Assay for cell-free protein synthesis- inhibiting activity --- p.121 / Chapter 3.3.2.5. --- Assay for ribonuclease activity --- p.121 / Chapter 3.3.2.6. --- Assay for N-glycosidase activity --- p.122 / Chapter 3.3.2.7. --- Assay for antifungal activity --- p.123 / Chapter 3.3.2.8. --- "Assessment of purity, activity and yield" --- p.124 / Chapter 3.3.3. --- Discussion --- p.125 / Chapter 3.4. --- Purification and Characterization of α and β Benincasin from fresh Winter Melon {Benincasa hispida var. dong-gua) Seeds / Chapter 3.4.1. --- Introduction --- p.127 / Chapter 3.4.2. --- Results --- p.129 / Chapter 3.4.2.1. --- Purification --- p.130 / Chapter 3.4.2.2. --- Molecular weight determination --- p.135 / Chapter 3.4.2.3. --- N-terminal amino acid sequence --- p.136 / Chapter 3.4.2.4. --- Assay for cell-free protein synthesis- inhibiting activity --- p.137 / Chapter 3.4.2.5. --- Assay for ribonuclease activity --- p.137 / Chapter 3.4.2.6. --- Assay for antifungal activity --- p.138 / Chapter 3.4.2.7. --- "Assessment of purity, activity and yield" --- p.140 / Chapter 3.4.3. --- Discussion --- p.141 / Chapter 3.5. --- Purification and characterization of Moschins from Pumpkin (Cucurbita moschata) Seeds / Chapter 3.5.1. --- Introduction --- p.143 / Chapter 3.5.2. --- Results --- p.145 / Chapter 3.5.2.1. --- Purification --- p.146 / Chapter 3.5.2.2. --- Molecular weight determination --- p.149 / Chapter 3.5.2.3. --- N-terminal amino acid sequence --- p.150 / Chapter 3.5.2.4. --- Assay for cell-free protein synthesis- inhibiting activity --- p.151 / Chapter 3.5.2.5. --- Assay for ribonuclease activity --- p.151 / Chapter 3.5.2.6. --- "Assessment of purity, activity and yield" --- p.152 / Chapter 3.5.3. --- Discussion --- p.153 / Chapter Chapter 4 --- General Discussion and Conclusion --- p.154 / References --- p.158

Tin-hung Ho. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1990. / Bibliography: leaves 137-151. / ACKNOWLEDGEMENT --- p.1 / CONTENT --- p.3 / ABBREVIATIONS --- p.7 / ABSTRACT --- p.8 / Chapter CHAPTER 1 --- INTRODUCTION --- p.10 / Chapter CHAPTER 2 --- LITERATURE REVIEW / Chapter 2.1 --- The cervix and cervical cancer --- p.12 / Chapter 2.2 --- Human papillomaviruses --- p.23 / Chapter 2.3 --- Culture of cancer cells --- p.36 / Chapter 2.4 --- Methods for the detection of HPV infection --- p.40 / Chapter CHAPTER 3 --- MATERIALS AND METHODS / Chapter 3.1 --- Characterization of cervical carcinoma cell lines / Chapter 3.1.1 --- Materials and solutions --- p.47 / Chapter 3.1.2 --- Establishment of cervical carcinoma cell lines --- p.50 / Chapter 3.1.3 --- Morphological studies of cervical carcinoma cells --- p.52 / Chapter 3.1.4 --- Examination of cervical carcinoma cells cultured on collagen gel --- p.54 / Chapter 3.1.5 --- Growth kinetics study --- p.55 / Chapter 3.1.6 --- Plating efficiency test --- p.56 / Chapter 3.1.7 --- Spheroid formation assay --- p.56 / Chapter 3.1.8 --- Chromosome number study --- p.57 / Chapter 3.2 --- Immunocytochemical studies / Chapter 3.2.1 --- Materials and solutions --- p.58 / Chapter 3.2.2 --- Immunocytochemical test for keratin --- p.59 / Chapter 3.2.3 --- Test for HPV capsid antigens --- p.60 / Chapter 3.3 --- Molecular studies of HPV in cervical carcinoma cells / Chapter 3.3.1 --- Materials and solutions --- p.62 / Chapter 3.3.2 --- Preparation of HPV DNA probes --- p.66 / Chapter 3.3.3 --- DNA extraction from the cervical carcinoma cells --- p.74 / Chapter 3.3.4 --- Detection of HPV DNA sequences by the use of hybridization techniques --- p.76 / Chapter 3.3.5 --- Copy number and physical state studies of HPV --- p.81 / Chapter 3.3.6 --- Study of the transcriptional activity of HPV DNA in cultured cervical carcinoma cells --- p.83 / Chapter CHAPTER 4 --- RESULTS / Chapter 4.1 --- Characterization of cervical carcinoma cell lines / Chapter 4.1.1 --- Morphological studies --- p.89 / Chapter 4.1.2 --- Examination of cervical carcinoma cells cultured on collagen gel --- p.90 / Chapter 4.1.3 --- Growth kinetics study --- p.93 / Chapter 4.1.4 --- Plating efficiency test --- p.94 / Chapter 4.1.5 --- Spheroid formation assay --- p.95 / Chapter 4.1.6 --- Chromosome number study --- p.98 / Chapter 4.2 --- Immunocytochemical studies / Chapter 4.2.1 --- Immunocytochemical test for keratin --- p.99 / Chapter 4.2.2 --- Test for HPV capsid antigen --- p.99 / Chapter 4.3 --- Molecular studies of HPV in cervical carcinoma cell lines / Chapter 4.3.1 --- Preparation of HPV DNA probes --- p.101 / Chapter 4.3.2 --- Detection of HPV DNA by the use of hybridization techniques --- p.102 / Chapter 4.3.3 --- Copy number and physical state studies --- p.105 / Chapter 4.3.4 --- Analysis of the transcriptional activity --- p.108 / Chapter CHAPTER 5 --- DISCUSSIONS / Chapter 5.1 --- Characterization of cervical carcinoma cell lines / Chapter 5.1.1 --- Morphological features of two cervical carcinoma cell lines --- p.110 / Chapter 5.1.2 --- Other characteristics of the cell lines --- p.112 / Chapter 5.2 --- Immunocytochemical studies / Chapter 5.2.1 --- Test for keratin antigens --- p.117 / Chapter 5.2.2 --- Test for HPV capsid antigens --- p.117 / Chapter 5.3 --- Molecular studies of HPV in cervical carcinoma cell lines / Chapter 5.3.1 --- Establishment of methods --- p.121 / Chapter 5.3.2 --- Detection of HPV DNA sequences by nucleic acid hybridizations --- p.123 / Chapter 5.3.3 --- Copy number and physical state studies --- p.128 / Chapter 5.3.4 --- Transcriptional analysis of HPV DNA in cervical carcinoma cell lines --- p.132 / CONCLUSION --- p.134 / REFERENCES --- p.137 / ILLUSTRATIONS --- p.152

by Mo Flora. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1991. / Includes bibliographical references (leaves 168-184). / ACKNOWLEDGEMENT --- p.i / ABSTRACT --- p.ii / TABLE OF CONTENTS --- p.iv / Chapter CHAPTER ONE --- INTRODUCTION / Chapter 1.1 --- Stroke as a major disabling disease --- p.1 / Chapter 1.2 --- Classification of stroke --- p.4 / Chapter 1.3 --- Risk factors attributing to stroke --- p.15 / Chapter 1.4 --- Experimental methods to induce cerebral ischemia --- p.19 / Chapter 1.4.1 --- The establishment of animal models for stroke --- p.21 / Chapter 1.4.2 --- Gerbil as a putative model --- p.25 / Chapter 1.5 --- Mechanisms of focal ischemia damage --- p.30 / Chapter 1.6 --- Potential biochemical markers for cerebral ischemia --- p.38 / Chapter 1.7 --- Aim of investigation --- p.48 / Chapter CHAPTER TWO --- MATERIALS AND METHODS / Chapter 2.1 --- Common chemicals --- p.49 / Chapter 2.2 --- Common bench solutions --- p.52 / Chapter 2.3 --- Animals / Chapter 2.3.1 --- Gerbils --- p.52 / Chapter 2.3.2 --- Rabbit --- p.53 / Chapter 2.4 --- Establishment of an animal model / Chapter 2.4.1 --- Surgical methods for common carotid artery (CCA) ligation --- p.54 / Chapter 2.5 --- Methods to determine stroke conditions of gerbils / Chapter 2.5.1 --- Ocular fundus examination --- p.56 / Chapter 2.5.2 --- Stroke index --- p.56 / Chapter 2.5.3 --- Inclined plane method --- p.59 / Chapter 2.6 --- Preparation of gerbil brain for subsequent analysis / Chapter 2.6.1 --- Preparation of gerbil brain slices --- p.61 / Chapter 2.6.2 --- "2,3,5-triphenytetrazolium chloride (TTC) for quantitative staining of brain slices" --- p.61 / Chapter 2.6.3 --- Preparation of normal and stroke gerbil brain extract --- p.62 / Chapter 2.7 --- Polyacrylamide gel electrophoresis (PAGE) using a discontinuous buffer system / Chapter 2.7.1 --- Stock reagents --- p.63 / Chapter 2.7.2 --- Separation gel preparation --- p.65 / Chapter 2.7.3 --- Stacking gel preparation --- p.66 / Chapter 2.7.4 --- Electrophoresis conditions --- p.67 / Chapter 2.7.5 --- Staining and destaining --- p.67 / Chapter 2.8 --- Two dimensional slab gel electrophoresis / Chapter 2.8.1 --- Equipment --- p.70 / Chapter 2.8.2 --- Chemical --- p.70 / Chapter 2.8.3 --- Procedure --- p.74 / Chapter 2.9 --- Production of rabbit polyclonal antibodies against isolated stroke protein / Chapter 2.9.1 --- Isolation of stroke protein band from SDS-PAGE slab gel --- p.78 / Chapter 2.9.2 --- Production of anti-stroke protein serum in rabbits --- p.79 / Chapter 2.10 --- Western blotting method / Chapter 2.10.1 --- Reagents --- p.80 / Chapter 2.10.2 --- Procedures --- p.81 / Chapter 2.11 --- Extraction of total cellular RNA by lithium chloride method / Chapter 2.11.1 --- Reagents --- p.83 / Chapter 2.11.2 --- Procedures --- p.84 / Chapter 2.11.3 --- Checking the purity of the extracted RNA --- p.85 / Chapter 2.12 --- Purification of mRNA / Chapter 2.12.1 --- Reagents --- p.85 / Chapter 2.12.2 --- Procedure --- p.86 / Chapter 2.13 --- Verification of purity of mRNA / Chapter 2.13.1 --- Reagents --- p.87 / Chapter 2.13.2 --- Procedure --- p.88 / Chapter 2.14 --- Translation of gerbil brain mRNA in reticulocyte lysates and analysis of its product by SDS PAGE / Chapter 2.14.1 --- Reagents --- p.89 / Chapter 2.14.2 --- Procedures --- p.89 / Chapter CHAPTER THREE --- ESTABLISHMENT OF AN ANIMAL STROKE MODEL / Chapter 3.1 --- Foreword --- p.92 / Chapter 3.2 --- Preliminary studies / Chapter 3.2.1 --- Introduction --- p.92 / Chapter 3.2.2 --- Results --- p.93 / Chapter 3.2.3 --- Discussion --- p.96 / Chapter 3.3 --- Survival rate analysis / Chapter 3.3.1 --- Introduction --- p.97 / Chapter 3.3.2 --- Result --- p.98 / Chapter 3.3.3 --- Discussion --- p.102 / Chapter 3.4 --- Neurologic signs of ischemia / Chapter 3.4.1 --- Introduction --- p.103 / Chapter 3.4.2 --- Result --- p.105 / Chapter 3.4.3 --- Discussion --- p.111 / Chapter 3.5 --- Ocular fundus examination / Chapter 3.5.1 --- Introduction --- p.112 / Chapter 3.5.2 --- Result --- p.114 / Chapter 3.5.3 --- Discussion --- p.116 / Chapter 3.6 --- Inclined plane method / Chapter 3.6.1 --- Introduction --- p.117 / Chapter 3.6.2 --- Result --- p.118 / Chapter 3.6.3 --- Discussion --- p.121 / Chapter 3.7 --- Histologic examination using TTC as staining agent / Chapter 3.7.1 --- Introduction --- p.122 / Chapter 3.7.2 --- Result --- p.124 / Chapter 3.7.3 --- Discussion --- p.129 / Chapter CHAPTER FOUR --- IDENTIFICATION OF ALTERED PROTEIN PATTERN IN THE - BRAINS OF STROKE GERBILS BY ELECTROPHORETIC METHODS / Chapter 4.1 --- Separation of soluble brain extracts by SDS-PAGE analysis / Chapter 4.1.1 --- Introduction --- p.130 / Chapter 4.1.2 --- Result --- p.132 / Chapter 4.1.3 --- Discussion --- p.140 / Chapter 4.2 --- Two dimensional electrophoretic analysis of soluble brain extracts from stroke gerbils / Chapter 4.2.1 --- Introduction --- p.142 / Chapter 4.2.2 --- Result --- p.143 / Chapter 4.2.3 --- Discussion --- p.148 / Chapter CHAPTER FIVE --- ISOLATION OF STROKE-ASSOCIATED PROTEIN FROM BRAINS OF STROKE GERBILS BY IMMUNOCHEMICAL METHOD / Chapter 5.1 --- Introduction --- p.149 / Chapter 5.2 --- Result --- p.151 / Chapter 5.3 --- Discussion --- p.153 / Chapter CHAPTER SIX --- DETECTION OF NEW PROTEIN TRANSLATED FROM MESSENGER RIBONUCLEIC ACID FROM BRAINS OF STROKE GERBIL / Chapter 6.1 --- Introduction / Chapter 6.1.1 --- Extraction of stroke gerbil brain messenger ribonucleic acid --- p.154 / Chapter 6.1.2 --- Translation of mRNA --- p.154 / Chapter 6.2 --- Results / Chapter 6.2.1 --- Yield of total cellular RNA --- p.157 / Chapter 6.2.2 --- Verification of purity of mRNA --- p.157 / Chapter 6.2.3 --- Autoradiographic patterns of translated proteins --- p.159 / Chapter 6.3 --- Discussion --- p.163 / Chapter CHAPTER SEVEN --- GENERAL DISCUSSION --- p.165 / BIBLIOGRAPHY --- p.168

Wong Pak Ki. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 86-88). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.iv / Dedication --- p.v / Table of Contents --- p.vi / List of Abbreviations --- p.ix / List of Appendices --- p.xi / List of Figures --- p.xiv / List of Tables --- p.xx / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1 --- Ginseng and Ginsenosides --- p.1 / Chapter 1.2 --- Instrumental Analysis of Ginsenosides --- p.6 / Chapter 1.2.1 --- Thin Layer Chromatography --- p.6 / Chapter 1.2.2 --- Infrared Spectroscopy --- p.7 / Chapter 1.2.3 --- Colorimetry --- p.7 / Chapter 1.2.4 --- Gas Chromatography --- p.7 / Chapter 1.2.5 --- High Performance Liquid Chromatography --- p.8 / Chapter 1.3 --- Objective of the Study --- p.9 / Chapter Chapter 2: --- Experimental --- p.13 / Chapter 2.1 --- History of Electrophoresis and Capillary Electrophoresis --- p.13 / Chapter 2.1.1 --- Electroosmotic Flow (EOF) --- p.14 / Chapter 2.1.2 --- Electrophoretic Migration --- p.18 / Chapter 2.2 --- Reagents and Materials --- p.20 / Chapter 2.2.1 --- Reagents and Glassware --- p.20 / Chapter 2.2.2 --- Instrumentation --- p.20 / Chapter 2.2.3 --- Preparation of Solutions and Wavelength Selection --- p.22 / Chapter 2.2 --- Procedures --- p.23 / Chapter Chapter 3: --- Results and Discussions --- p.24 / Chapter 3.1 --- Initial Selection of the Running Electrolyte --- p.24 / Chapter 3.2 --- Inclusion Additives in the Aqueous Buffer Solution --- p.29 / Chapter 3.2.1 --- Reasons for Addition of Buffer Additives --- p.29 / Chapter 3.2.1.1 --- Cyclodextrin --- p.29 / Chapter 3.3 --- Addition of Surfactants --- p.33 / Chapter 3.3.1 --- Sodium Dodecyl Sulfate (SDS) --- p.35 / Chapter 3.3.2 --- Sodium Cholate --- p.41 / Chapter 3.4 --- Addition of Organic Modifier --- p.43 / Chapter 3.5 --- Effect of pH --- p.46 / Chapter 3.6 --- Effect of the Concentration of the Borate/Phosphate Solution --- p.51 / Chapter 3.7 --- Effect of Capillaries with Different Inner Diameters (I.D.) --- p.54 / Chapter 3.7.1 --- Effect of pH --- p.54 / Chapter 3.7.2 --- Effect of the Buffer Concentration --- p.60 / Chapter 3.7.3 --- Comparison of Migration Time between Capillaries of 50μm and 75μm Inner Diameter --- p.62 / Chapter 3.8 --- Optimization of Other Experimental Parameters --- p.66 / Chapter 3.8.1 --- Applied Voltage --- p.66 / Chapter 3.8.2 --- The Time of Injection --- p.68 / Chapter 3.8.3 --- The Operating Temperature --- p.70 / Chapter 3.9 --- Intra-day and Inter-day Reproducibility --- p.72 / Chapter 3.10 --- Quantitative Analysis of the Ginsenosides --- p.74 / Chapter 3.11 --- Application of the Developed Methodology --- p.78 / Chapter 3.11.1 --- Experimental Procedures --- p.79 / Chapter Chapter 4: --- Conclusion --- p.83 / References --- p.86 / Appendices --- p.89

by Cheung Yuk Yin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves xx-xxviii). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 摘要 --- p.iii / Table of Content --- p.iv / Abbreviations / for Amino Acids --- p.viii / for Standard Genetic Code --- p.ix / for Units --- p.x / for Prefixes --- p.xi / for Terms commonly used in the report --- p.xii / List of Figures --- p.xiii / List of Tables --- p.xiv / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Epidemiology of Hepatitis B Virus (HBV) --- p.1 / Chapter 1.2 --- Relationship between Hepatitis B Virus and Hepatocellular Carcinoma --- p.2 / Chapter 1.3 --- Brief Description of HBV Genome --- p.2 / Chapter 1.4 --- Possible Roles of HBx in Hepatocellular Carcinoma --- p.4 / Chapter 1.5 --- Novel Interacting Partner of HBx - HBx-lnteracting Protein (XIP) --- p.6 / Chapter 1.6 --- Objective --- p.6 / Chapter Chapter 2 --- Methodology / Chapter 2.1 --- Information of the HBx and XIP Clones --- p.7 / Chapter 2.2 --- "Information of the Expression Vectors (pRSETA, 6xHis-pRSETA and pET8C)" --- p.7 / Chapter 2.3 --- Sub-Cloning of HBx and XIP into Different Vectors --- p.9 / Chapter 2.3.1 --- Design of Primers for Cloning of HBx and XIP into Different Vectors --- p.9 / Chapter 2.3.2 --- Polymerase Chain Reaction (PCR) Protocol --- p.12 / Chapter 2.3.3 --- Enzyme Digestion Reaction Protocol --- p.14 / Chapter 2.3.4 --- Ligation Protocol --- p.16 / Chapter 2.3.5 --- Preparation of Competent Cells --- p.17 / Chapter 2.3.6 --- Transformation --- p.18 / Chapter 2.3.7 --- Gel Extraction Protocol --- p.19 / Chapter 2.3.7.1 --- Life Technologies CONCERT´ёØ Rapid Gel Extraction System --- p.19 / Chapter 2.3.7.2 --- QIAGEN Gel Extraction Kit --- p.20 / Chapter 2.3.8 --- Plasmid Preparation Protocol --- p.22 / Chapter 2.3.8.1 --- Life Technologies CONCERT´ёØ Rapid Plasmid Minipreps --- p.22 / Chapter 2.3.8.2 --- QIAGEN Plasmid Maxi Kit --- p.23 / Chapter 2.4 --- Expression of HBx and XIP in E. coli Strain C41 (DE3) --- p.25 / Chapter 2.4.1 --- Transformation --- p.25 / Chapter 2.4.2 --- Expression of HBx and 6xHis-HBx in E. coli Strain C41 (DE3) --- p.26 / Chapter 2.4.3 --- Expression of XIP in E. coli Strain C41 (DE3) --- p.27 / Chapter 2.5 --- Preparation of Buffers for Chromatography and Circular Dichroism Spectrum Measurement --- p.28 / Chapter 2.6 --- Purification and Refolding of HBx and His-Tagged HBx --- p.28 / Chapter 2.6.1 --- Washing of HBx and His-Tagged HBx Inclusion Bodies --- p.28 / Chapter 2.6.2 --- His-Tagged HBx Purification by Affinity Chromatography --- p.29 / Chapter 2.6.3 --- HBx Purification by Size Exclusion Chromatography --- p.30 / Chapter 2.6.4 --- Refolding of HBx and His-Tagged HBx by Oxidative Dialysis --- p.30 / Chapter 2.7 --- Purification of XIP --- p.33 / Chapter 2.7.1 --- Screening of Chromatographic Conditions for the Purification of XIP --- p.33 / Chapter 2.7.2 --- XIP 1st Step of Purification by Hydrophobic Interaction Chromatography --- p.34 / Chapter 2.7.3 --- XIP 2nd step of Purification by Size Exclusion Chromatography --- p.34 / Chapter 2.8 --- Chemical Denaturation Experiment of HBx and XIP --- p.36 / Chapter 2.8.1 --- Preparation of Urea Buffers for the Chemical Denaturation of HBx --- p.37 / Chapter 2.8.2 --- Preparation of Different GdnHCI Buffer for the Chemical Denaturation of XIP --- p.38 / Chapter 2.8.3 --- Calculation for Chemical Denaturation Experiment --- p.39 / Chapter 2.8.3.1 --- Protein Concentration Calculation --- p.39 / Chapter 2.8.3.2 --- Residual Molar Elipticity Calculation --- p.39 / Chapter 2.8.3.3 --- Free Energy Change (ΔGu) Calculation --- p.40 / Chapter 2.9 --- Two-dimensional Heteronuclear Nuclear Magnetic Resonance (NMR) Experiment --- p.41 / Chapter 2.10 --- Interaction Confirmation between HBx and XIP --- p.42 / Chapter 2.10.1 --- "Transfection of pEGFP, pEGFP-HBx and pEGFP-XIP into HepG2" --- p.42 / Chapter 2.10.2 --- Yeast Two Hybrid System for Confirmation of HBx and XIP Interaction --- p.44 / Chapter 2.10.2.1 --- Preparation of Y187 Competent Cells --- p.44 / Chapter 2.10.2.2 --- Transformation of pGBKT7-HBx and pACT2-XIP into Y187 --- p.45 / Chapter 2.10.2.3 --- β-galactosidase Colony Lift Assay --- p.46 / Chapter Chapter 3 --- "Expression, Purification and Characterization of Hepatitis B Virus X Protein (HBx)" / Chapter 3.1 --- Introduction --- p.47 / Chapter 3.2 --- Construction of Recombinant HBx-pRSETA and 6xHis-HBx-pRSETA Plasmids --- p.48 / Chapter 3.3 --- Expression of 6xHis-HBx in E. coli C41 (DE3) using M9ZB Medium --- p.52 / Chapter 3.4 --- Expression of HBx in E. coli C41 (DE3) using M9ZB Medium --- p.54 / Chapter 3.5 --- Purification and Refolding of 6xHis-HBx Fusion Proteins --- p.56 / Chapter 3.6 --- Purification and Refolding of HBx Proteins --- p.60 / Chapter 3.7 --- Structural Characterization of Refolded HBx --- p.65 / Chapter 3.7.1 --- Introduction --- p.55 / Chapter 3.7.2 --- Experimental Analysis of HBx Secondary Structure --- p.66 / Chapter 3.7.3 --- Chemical Unfolding Experiment of HBx --- p.68 / Chapter 3.8 --- Discussion --- p.70 / Chapter 3.8.1 --- "HBx was Expressed, Purified and Characterized instead of 6xHis-HBx" --- p.71 / Chapter 3.8.2 --- High Concentration of DTT was used to Minimize Formation of HBx Aggregates --- p.72 / Chapter 3.8.3 --- Oxidative Refolding to Ensure Proper Disulfide Bond Formation --- p.73 / Chapter 3.8.4 --- Computational Prediction and Experimental Prediction of Secondary Structure of HBx --- p.75 / Chapter 3.9 --- Concluding Remarks --- p.77 / Chapter Chapter 4 --- "Expression, Purification and Characterization of HBx-lnteracting Protein (XIP)" / Chapter 4.1 --- Introduction --- p.78 / Chapter 4.2 --- Construction of Recombinant XIP-pET8C --- p.78 / Chapter 4.3 --- Expression of XIP in E. coli C41 (DE3) using M9ZB and M9 Mediums --- p.82 / Chapter 4.4 --- Screening of Chromatographic Conditions for the Purification of XIP --- p.83 / Chapter 4.4.1 --- Introduction --- p.83 / Chapter 4.4.2 --- Purification Details --- p.83 / Chapter 4.5 --- Purification of XIP by HiTrap Phenyl HP 5-ml Column --- p.87 / Chapter 4.6 --- Purification of XIP by HiLoad 26/60 Superdex 75 Prep Grade --- p.89 / Chapter 4.7 --- Structural Characterization of XIP --- p.92 / Chapter 4.7.1 --- CD Spectrum --- p.92 / Chapter 4.7.2 --- Chemical Denaturation Experiment of XIP --- p.93 / Chapter 4.7.3 --- Two-Dimensional Heteronuclear Nuclear Magnetic Resonance (NMR) Spectrum of 15N Labeled XIP --- p.95 / Chapter 4.8 --- Discussion --- p.97 / Chapter 4.8.1 --- Purification Method Development --- p.97 / Chapter 4.8.2 --- "Do Different Protein Cosolutes, Protein Stabilizers and Detergents Help XIP to Adopt a Stable Conformation?" --- p.99 / Chapter 4.9 --- Concluding Remarks --- p.101 / Chapter Chapter 5 --- In vivo Studies of HBx and XIP Interactions / Chapter 5.1 --- Investigation of Sub-Cellular Localization of HBx and XIP in Liver Cells --- p.102 / Chapter 5.1.1 --- Introduction --- p.102 / Chapter 5.1.2 --- "Construction of Recombinant HBx-pECFP-C1, HBx-pEGFP-C1, HBx-pEYFP-C1 and XIP-pECFP-C1, XIP-pEGFP-C1, XIP-pEYFP-C1" --- p.103 / Chapter 5.1.3 --- Transfection of pEGFP-C1 HBx and pEGFP-C1 XIP into HepG2 to Find Out HBx and XIP Sub-Cellular Localization --- p.106 / Chapter 5.1.3.1 --- Introduction --- p.107 / Chapter 5.1.3.2 --- Investigation of EGFP Proteins Expression using the Confocal Microscope and the Leica TCS Software --- p.108 / Chapter 5.1.4 --- Discussion and Future Prospects --- p.111 / Chapter 5.2 --- Interaction of HBx and XIP Studied by Yeast Two-Hybrid System --- p.113 / Chapter 5.2.1 --- Introduction --- p.113 / Chapter 5.2.2 --- Construction of Recombinant HBx-pGBKT7 and XIP-pACT2 Plasmids --- p.114 / Chapter 5.2.3 --- Confirmation of HBx and XIP Interaction by Yeast Two-Hybrid System --- p.117 / Chapter 5.2.4 --- Discussion --- p.121 / Chapter Chapter 6 --- Conclusion --- p.123 / Appendix I Sequence of HBx and XIP --- p.I / Chapter II --- Vector Sequences --- p.II / Chapter III --- Vector Maps --- p.VI / Chapter IV --- Electrophoresis Markers --- p.XI / Chapter V --- Agarose Gel Electrophoresis --- p.XII / Chapter VI --- SDS-PAGE Eectrophoresis --- p.XIII / Chapter VII --- Medium for Bacterial Culture --- p.XV / Chapter VIII --- Medium for Cell Culture --- p.XVII / Chapter IX --- Medium for Yeast Culture --- p.XVIII / Chapter X --- Buffers for Yeast Transformation --- p.XIX / Reference --- p.XX

Ip Yee-man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 85-89). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Abstract --- p.iii / Table of Contents --- p.vi / Abbreviations --- p.xi / List of Figures --- p.xiii / List of Tables --- p.xvii / Chapter / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Background --- p.1 / Chapter 1.2 --- Quality control of Chinese herbal medicine --- p.2 / Chapter 1.3 --- Rhizoma Chuanxiong --- p.4 / Chapter 1.3.1. --- General description --- p.4 / Chapter 1.3.2. --- Chemical constituents --- p.4 / Chapter 1.3.3. --- Pharmacology --- p.7 / Chapter 1.3.4 --- Instrumental analysis --- p.9 / Chapter 1.3.4.1 --- Thin Layer Chromatography (TLC) --- p.9 / Chapter 1.3.4.2 --- Gas Chromatography (GC) --- p.10 / Chapter 1.3.4.3 --- High Performance Liquid Chromatography (HPLC) --- p.10 / Chapter 1.3.4.4 --- Capillary Electrophoresis (CE) --- p.10 / Chapter 1.4 --- Objectives of the study --- p.11 / Chapter 2. --- "Isolation, Characterization and Identification of Reference Compounds" --- p.13 / Chapter 2.1 --- General experiment procedures --- p.13 / Chapter 2.1.1. --- Solvents for chromatographic separation --- p.13 / Chapter 2.1.2 --- Chromatographic methods --- p.13 / Chapter 2.1.2.1 --- Adsorption column chromatography --- p.13 / Chapter 2.1.2.2 --- Thin layer chromatography --- p.13 / Chapter 2.1.2.3 --- Preparative layer chromatography --- p.14 / Chapter 2.1.3 --- Determination of physical data --- p.14 / Chapter 2.1.3.1 --- Infrared (IR) absorption spectra --- p.14 / Chapter 2.1.3.2 --- Nuclear Magnetic Resonance (NMR) spectra --- p.14 / Chapter 2.1.3.3 --- Mass spectra (MS) --- p.15 / Chapter 2.1.3.4 --- X-ray crystallography --- p.15 / Chapter 2.1.4 --- Authentic reference compounds --- p.15 / Chapter 2.2 --- "Procurement, extraction and initial fractionation of Rhizoma Chuanxiong" --- p.15 / Chapter 2.3 --- Chromatographic separation of the chloroform extract --- p.16 / Chapter 2.3.1 --- Chromatographic separation of fraction F1002 --- p.16 / Chapter 2.3.1.1 --- Spectral data for the characterization of compound 1 [5-(hydroxymethyl)- 2- furancarboxaldehyde] --- p.17 / Chapter 2.3.2 --- Column chromatographic separation of fraction F1003A --- p.17 / Chapter 2.3.2.1 --- Spectral data for the characterization of compound 2 (oleic acid) --- p.18 / Chapter 2.3.2.2 --- Physical data for the characterization of compound 3 (ferulic acid) --- p.18 / Chapter 2.3.3 --- Preparative layer chromatographic separation of Fraction F1010 --- p.19 / Chapter 2.3.3.1 --- Spectral data for the characterization of compound 4 (daucosterol) --- p.19 / Chapter 2.4 --- Column chromatographic separation of the hexane extract --- p.19 / Chapter 2.4.1 --- Removal of fatty acids in fraction F2005 and F2006 by partition --- p.20 / Chapter 2.4.2 --- Column chromatographic separation of fraction F2005M --- p.20 / Chapter 2.4.2.1 --- Spectral data for the characterization of compound 5 (butylidenephthalide) --- p.20 / Chapter 2.4.2.2 --- Spectral data for the characterization of compound 6 (butylphthalide) --- p.21 / Chapter 2.4.3 --- Column chromatographic separation of fraction F2006M --- p.21 / Chapter 2.4.3.1 --- "Spectral data for the characterization of compound 7 (Z, Z'-6.6', 7.3'a- diligustilide)" --- p.21 / Chapter 2.4.4 --- Colum chromatographic separation of fraction --- p.22 / Chapter 2.4.4.1 --- Spectral data for the characterization of compound 8 (pregnenolone) --- p.22 / Chapter 2.4.4.2 --- "Spectral data for the characterization of compound 9 [5,5- oxydimethylenebis(2-furaldehyde)]" --- p.23 / Chapter 2.5 --- Results and Discussion --- p.24 / Chapter 2.5.1 --- Identification of compound 1 [5-(hydroxymethyl)-2- furancarboxaldehyde] --- p.24 / Chapter 2.5.2 --- Identification of compound 2 (oleic acid) --- p.25 / Chapter 2.5.3 --- Identification of compound 3 (ferulic acid) --- p.26 / Chapter 2.5.4 --- Identification of compound 4 (daucosterol) --- p.26 / Chapter 2.5.5 --- Identification of compound 5 (butylidenephthalide) --- p.27 / Chapter 2.5.6 --- Identification of compound 6 (butylphthalide) --- p.28 / Chapter 2.5.7 --- "Identification of compound 7 (Z, Z'-6.6', 7.3'a-diligustilide)" --- p.30 / Chapter 2.5.8 --- Identification of compound 8 (pregnenolone) --- p.31 / Chapter 2.5.9 --- "Identification of compound 9 [5,5'-oxydimethylenebis(2-furaldehyde)]" --- p.32 / Chapter 2.6 --- Conclusions --- p.34 / Chapter 3. --- Analysis of Rhizoma Chuanxiong by Capillary Electrophoresis --- p.35 / Chapter 3.1 --- Introduction --- p.35 / Chapter 3.1.1 --- Capillary electrophoreis system --- p.35 / Chapter 3.1.2 --- Principles of separation --- p.36 / Chapter 3.1.3 --- Considerations on development of analysis method --- p.41 / Chapter 3.2 --- Experimental --- p.43 / Chapter 3.2.1 --- Reagents and materials --- p.43 / Chapter 3.2.2 --- Reference compounds --- p.43 / Chapter 3.2.3 --- Instrumentation and apparatus --- p.44 / Chapter 3.2.4 --- Experimental procedures --- p.45 / Chapter 3.2.4.1 --- Preparation of running buffer solution --- p.45 / Chapter 3.2.4.2 --- Preparation of standard solutions --- p.46 / Chapter 3.2.4.3 --- Preparation of Rhizoma Chuanxiong extracts --- p.47 / Chapter 3.2.4.4 --- Flushing of capillaries --- p.47 / Chapter 3.2.4.5 --- Conditions of separation --- p.48 / Chapter 3.3 --- Results and Discussion --- p.48 / Chapter 3.3.1 --- Preliminary experiments --- p.48 / Chapter 3.3.1.1 --- Addition of surfactants --- p.51 / Chapter 3.3.2 --- Effect of buffer concentration --- p.54 / Chapter 3.3.3 --- Effect of SDS concentration --- p.59 / Chapter 3.3.4 --- Addition of organic modifier --- p.63 / Chapter 3.3.5 --- Reproducibility of the proposed method --- p.68 / Chapter 3.3.6 --- Quantitative analysis of seven standard compounds --- p.70 / Chapter 3.3.7 --- Application of the developed methodology --- p.74 / Chapter 3.3.8 --- Conclusions --- p.83 / References --- p.85 / Appendices / Appendix 1.1.1 1H-NMR spectrum of 5-(hydroxymethyl)-2-furancarboxaldehyde --- p.90 / Appendix 1.1.2 13C-NMR spectrum of 5-(hydroxyinethyl)-2-furancarboxaldehyde --- p.90 / Appendix 1.2 X-ray crystallographic data of ferulic acid --- p.91 / Appendix 1.3 13C-NMR spectrum of butylidenephthalide --- p.96 / Appendix 1.4.1 1 H-NMR spectrum of butylphthalide --- p.97 / Appendix 1.4.2 13C-NMR spectrum of butylphthalide --- p.97 / "Appendix 1.5 X-ray crystallographic data of z, z', 6.6'， 7.3'a-diligustilide" --- p.98 / "Appendix 1.6 X-ray crystallographic data of 5,5'-oxydimethylenebis(2-furaldehyde)" --- p.105 / Appendix 2.1 Details of quantitative analysis of 5-(hydroxymethyl)-2-furancarboxaldehyde --- p.112 / Appendix 2.2 Details of quantitative analysis of ligustrazin hydrochloride --- p.112 / "Appendix 2.3 Details of quantitative analysis of 5,5'-oxydimethylenebis(2-furaldehyde)" --- p.113 / Appendix 2.4 Details of quantitative anlaysis of ferulic acid --- p.113 / Appendix 2.5 Details of quantitative analysis of butylphthalide --- p.114 / Appendix 2.6 Details of quantitative analysis of butylidenephthalide --- p.114 / "Appendix 2.7 Details of quantitative anlaysis of z,z', 6.6', 7.3'a-diligustilide" --- p.115 / Appendix 3.1 Quantitative analysis of Chuanxiong sample from Hong Kong (HK1) --- p.115 / Appendix 3.2 Quantitaive analysis of Chuanxiong sample from Hong Kong (HK2) --- p.116 / Appendix 3.3 Quantitative analysis of Chuanxiong sample from Sichuan (SC1) --- p.116 / Appendix 3.4 Quantitative analysis of Chuanxiong sample from Sichuan (SC2) --- p.117 / Appendix 3.5 Quantitative anlaysis of Chuanxiong samplefrom Fujian (FJ) --- p.117

by Ricardo Wong Chi Ho. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 128-138). / Abstracts in English and Chinese. / Acknowledgments --- p.i / Abstract --- p.ii / 論文摘要 --- p.iv / Table of Contents --- p.vi / List of Figures --- p.xi / List of Tables --- p.xiii / List of Abbreviations --- p.xiv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Overview of Serine Protease Inhibitors --- p.1 / Chapter 1.2 --- Classification of Serine Protease Inhibitors --- p.2 / Chapter 1.2.1 --- Kunitz Type Serine Protease Inhibitors --- p.7 / Chapter 1.2.2 --- Bowman-Birk Type Serine Protease Inhibitors --- p.11 / Chapter 1.2.3 --- Squash Type Serine Protease Inhibitors --- p.16 / Chapter 1.3 --- Role of Serine Protease Inhibitors in Plants --- p.20 / Chapter 1.4 --- Nutritional Fact of Serine Protease Inhibitors --- p.22 / Chapter 1.5 --- Possible Applications of Serine Protease Inhibitors --- p.25 / Chapter 1.5.1 --- Medical Applications --- p.25 / Chapter 1.5.2 --- Agricultural Applications --- p.29 / Chapter 1.6 --- Rationale of the Present Study --- p.31 / Chapter Chapter 2 --- Screening of Seeds for Inhibitory Activities Against Serine Proteases --- p.33 / Chapter 2.1 --- Introduction --- p.33 / Chapter 2.2 --- Materials and Methods --- p.37 / Chapter 2.2.1 --- Materials --- p.37 / Chapter 2.2.2 --- Extraction Method --- p.37 / Chapter 2.2.3 --- Assays for Proteases Inhibitory Activities --- p.38 / Chapter 2.2.3.1 --- Assay for Chymotrypsin Activity --- p.38 / Chapter 2.2.3.2 --- Assay for Trypsin Activity --- p.38 / Chapter 2.2.3.3 --- Assay for Elastase Activity --- p.39 / Chapter 2.2.3.4 --- Assay for Subtilisin Activity --- p.39 / Chapter 2.2.3.5 --- Assays for Protease Inhibitory Activities --- p.40 / Chapter 2.2.4 --- Determination of Protein Concentration --- p.41 / Chapter 2.3 --- Results --- p.42 / Chapter 2.3.1 --- Extraction --- p.42 / Chapter 2.3.2 --- Serine Proteases Inhibitory Activities --- p.42 / Chapter 2.4 --- Discussion --- p.47 / Chapter Chapter 3 --- Isolation of Chymotrypsin Inhibitor and Trypsin Inhibitors from Momordica cochinchinensis Seeds --- p.49 / Chapter 3.1 --- Introduction --- p.49 / Chapter 3.2 --- Materials and Methods --- p.56 / Chapter 3.2.1 --- Materials --- p.56 / Chapter 3.2.2 --- Protein Extraction --- p.57 / Chapter 3.2.3 --- SP-Sepharose Chromatography --- p.57 / Chapter 3.2.4 --- Reversed Phase High Pressure Liquid Chromatography --- p.58 / Chapter 3.2.5 --- Assays for Chymotrypsin and Trypsin Inhibitory Activities --- p.60 / Chapter 3.2.6 --- Titration of Chymotrypsin --- p.61 / Chapter 3.2.7 --- Tricine Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis --- p.62 / Chapter 3.2.8 --- Coupling of Trypsin-Sepharose 4B Affinity Column --- p.63 / Chapter 3.2.9 --- Affinity Chromatography on Trypsin-Sepharose 4B --- p.64 / Chapter 3.3 --- Results --- p.65 / Chapter 3.3.1 --- SP-Sepharose Chromatography --- p.65 / Chapter 3.3.2 --- Reversed Phase High Pressure Liquid Chromatography --- p.67 / Chapter 3.3.3 --- Summary of Purification --- p.71 / Chapter 3.3.4 --- Titration of Chymotrypsin --- p.74 / Chapter 3.3.5 --- Tricine Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis --- p.74 / Chapter 3.3.6 --- Affinity Chromatography on Trypsin-Sepharose 4B --- p.78 / Chapter 3.4 --- Discussion --- p.81 / Chapter Chapter 4 --- Characterization of Chymotrypsin Inhibitor and Trypsin Inhibitors --- p.88 / Chapter 4.1 --- Introduction --- p.88 / Chapter 4.2 --- Materials and Methods --- p.90 / Chapter 4.2.1 --- Materials --- p.90 / Chapter 4.2.2 --- Determination of Molecular Weight --- p.90 / Chapter 4.2.3 --- Amino Acid Sequence Analysis --- p.91 / Chapter 4.2.4 --- Surface Plasmon Resonance Measurement --- p.92 / Chapter 4.2.4.1 --- Immobilization of Ligands on the Surface of Optical Biosensors --- p.92 / Chapter 4.2.4.2 --- Determination of Kinetics Constants --- p.93 / Chapter 4.2.4.3 --- pH Dependence of the Inhibition by Chymotrypsin Inhibitor --- p.93 / Chapter 4.2.4.4 --- Data Analysis --- p.94 / Chapter 4.2.5 --- Effect of Chymotrypsin Inhibitor on the Estereolytic Activity and Proteolytic Activity of Chymotrypsin --- p.95 / Chapter 4.2.6 --- Specificities of the Inhibitors % --- p.96 / Chapter 4.2.7 --- Binding Ratio of CI to Different Proteases --- p.97 / Chapter 4.2.8 --- Effects of the Proteases on Their Corresponding Inhibitors --- p.97 / Chapter 4.2.8.1 --- Tricine Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis --- p.97 / Chapter 4.2.8.2 --- Assay for Chymotrypsin Inhibitory Activity --- p.98 / Chapter 4.3 --- Results --- p.99 / Chapter 4.3.1 --- Molecular Weight of the Inhibitors --- p.99 / Chapter 4.3.2 --- N-terminal Amino Acid Sequence --- p.99 / Chapter 4.3.3 --- Surface Plasmon Resonance Measurement --- p.102 / Chapter 4.3.3.1 --- Kinetics of Chymotrypsin Inhibitor --- p.102 / Chapter 4.3.3.2 --- Kinetics of Trypsin Inhibitors --- p.106 / Chapter 4.3.3.3 --- pH Dependence of the Inhibition by Chymotrypsin Inhibitor --- p.106 / Chapter 4.3.4 --- Effect of Chymotrypsin Inhibitor on the Estereolytic Activity and Proteolytic Activity of Chymotrypsin --- p.106 / Chapter 4.3.5 --- Specificities of the Inhibitors --- p.110 / Chapter 4.3.6 --- Binding Ratio of CI to Different Proteases --- p.112 / Chapter 4.3.7 --- Effects of the Proteases on Their Corresponding Inhibitors --- p.112 / Chapter 4.4 --- Discussion --- p.119 / Chapter Chapter 5 --- Conclusion --- p.125 / References --- p.128

by Choy Ka-Fai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 107-125). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.I / 論文摘要 --- p.II / ABSTRACT --- p.III / ABBREVIATIONS --- p.V / TABLE OF CONTENTS --- p.VI / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter CHAPTER 2 --- PURIFICATION OF GRASS CARP ALDH FROM MITOCHONDRIA --- p.18 / Chapter CHAPTER 3 --- PURIFICATION & CHARACTERIZATION OF GRASS CARP ALDH --- p.49 / Chapter CHAPTER 4 --- CONCLUSION --- p.104 / REFERENCES --- p.107

by Tang Wai Kwan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 91-110). / Abstracts in English and Chinese. / Acknowledgements / 論文摘要 / Abstract / Abbreviations / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Aldehyde Dehydrogenase Extended Family --- p.1 / Chapter 1.1.1 --- Phylogenetic Tree --- p.2 / Chapter 1.1.2 --- Physiological Functions --- p.4 / Chapter 1.1.3 --- Structural Conservations --- p.7 / Chapter 1.2 --- ALDH-1 and ALDH-2 --- p.9 / Chapter 1.3 --- Antiquitin --- p.11 / Chapter 1.4 --- Osmoregulation --- p.14 / Chapter 1.4.1 --- Osmoprotectant --- p.14 / Chapter 1.4.2 --- Betaine Aldehyde Dehydrogenase --- p.15 / Chapter 1.5 --- Objectives of the Present Study --- p.18 / Chapter Chapter 2 --- Purification and Characterization of Seabream ALDH-2 and Antiquitin --- p.20 / Chapter 2.1 --- Introduction --- p.20 / Chapter 2.2 --- Materials --- p.21 / Chapter 2.3 --- Methodology / Chapter 2.3.1 --- Preparation of Crude Tissue Extract --- p.22 / Chapter 2.3.2 --- Synthesis of α-Cyanocinnamate Sepharose --- p.22 / Chapter 2.3.3 --- Synthesis of p-Hydroxyacetophenone Sepharose --- p.23 / Chapter 2.3.4 --- Purification of ALDH-2 --- p.23 / Chapter 2.3.5 --- Purification of Antiquitin --- p.24 / Chapter 2.3.6 --- Enzyme and Protein Assays --- p.24 / Chapter 2.3.7 --- Electrophoretic Procedures / Chapter 2.3.7.1 --- Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.26 / Chapter 2.3.7.2 --- Native PAGE --- p.27 / Chapter 2.3.7.3 --- Isoelectric focusing (IEF) --- p.27 / Chapter 2.3.8 --- N-terminal Amino Acid Sequencing --- p.28 / Chapter 2.4 --- Results / Chapter 2.4.1 --- Tissue Distribution of ALDH --- p.29 / Chapter 2.4.2 --- Purification and Molecular Properties of ALDH-2 --- p.31 / Chapter 2.4.3 --- Kinetic Properties of ALDH-2 --- p.42 / Chapter 2.4.4 --- Purification and Molecular Properties of Antiquitin --- p.49 / Chapter 2.4.5 --- Kinetic Properties of Antiquitin --- p.54 / Chapter Chapter 3 --- Discussion / Chapter 3.1 --- Tissue Distribution --- p.66 / Chapter 3.2 --- N-terminal Amino Acid Sequencing --- p.67 / Chapter 3.3 --- Purification of Seabream ALDH --- p.68 / Chapter 3.3.1 --- Separation of Two ALDH isoforms --- p.69 / Chapter 3.3.2 --- Binding Affinity of α-Cyanocinnamate Sepharose --- p.70 / Chapter 3.3.3 --- Purification --- p.72 / Chapter 3.4 --- Electrophoretic Properties --- p.73 / Chapter 3.5 --- pH and Temperature Stability --- p.74 / Chapter 3.6 --- Substrate Specificity --- p.77 / Chapter 3.7 --- Possible Functions of Antiquitin --- p.80 / Chapter 3.8 --- Future Prospects --- p.84 / Chapter Chapter 4 --- Conclusion --- p.90 / Chapter Chapter 5 --- References --- p.91