Ho Hing Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 140-156). / Abstracts in English and Chinese. / Chapter Chapter 1 --- General introduction / Chapter 1.1 --- History of soybean --- p.1 / Chapter 1.2 --- Health benefits of soybean --- p.2 / Chapter 1.3 --- Introduction to flavonoids --- p.2 / Chapter 1.4 --- Bioavailability of flavonoids from foods --- p.3 / Chapter 1.5 --- Pharmacological effects of flavonoids and their glycosides --- p.4 / Chapter 1.5.1 --- Anticarcinogenic activity --- p.4 / Chapter 1.5.2 --- Antioxidative activity --- p.7 / Chapter 1.5.3 --- Cardioprotective activity --- p.9 / Chapter 1.5.4 --- Osteoprotective activity --- p.10 / Chapter 1.5.5 --- Neuroprotective activity --- p.12 / Chapter 1.5.6 --- Antiangiogenic activity --- p.12 / Chapter 1.6 --- Soy leaves --- p.13 / Chapter Chapter 2 --- Isolation and purification of kaempferol glycosides and genistin in soy leaves / Chapter 2.1 --- Introduction --- p.14 / Chapter 2.2 --- Objectives --- p.15 / Chapter 2.3 --- Materials and Methods --- p.16 / Chapter 2.3.1 --- Extraction and isolation --- p.16 / Chapter 2.3.1.1 --- Preparation of soy leaves butanol extract --- p.16 / Chapter 2.3.1.2 --- Preparation of kaempferol glycosides from soy leaves butanol extract --- p.16 / Chapter 2.3.2 --- High performance liquid chromatography (HPLC) analysis --- p.19 / Chapter 2.3.2.1 --- Sample preparation for the HPLC analysis --- p.19 / Chapter 2.3.2.2 --- HPLC analysis --- p.19 / Chapter 2.3.2.3 --- Quantification of the flavonoids and their glycosides --- p.23 / Chapter 2.3.2.4 --- Change in flavonoids and their glycosides in soy leaves --- p.23 / Chapter 2.4 --- Results --- p.24 / Chapter 2.4.1 --- Compound 1 --- p.24 / Chapter 2.4.2 --- Compound 2 --- p.24 / Chapter 2.4.3 --- Compound 3 --- p.25 / Chapter 2.4.4 --- Compound 4 --- p.25 / Chapter 2.4.5 --- Compound 5 --- p.25 / Chapter 2.4.6 --- Compound 6 --- p.26 / Chapter 2.4.7 --- Quantification of flavonoids in soybean and soy leaves --- p.32 / Chapter 2.4.8 --- Age-dependent changes in flavonoids and their glycosides --- p.32 / Chapter 2.5 --- Discussion --- p.35 / Chapter 2.5.1 --- Compound 1 --- p.35 / Chapter 2.5.2 --- Compound 2 --- p.35 / Chapter 2.5.3 --- Compound 3 --- p.37 / Chapter 2.5.4 --- Compound 4 --- p.38 / Chapter 2.5.5 --- Compound 5 --- p.39 / Chapter 2.5.6 --- Compound 6 --- p.40 / Chapter 2.5.7 --- Age-dependent changes in flavonoids and their glycosides --- p.40 / Chapter Chapter 3 --- Hypolipidemic effects of soy leaves in hamsters / Chapter 3.1 --- Introduction --- p.41 / Chapter 3.1.1 --- Different lipoproteins and their functions --- p.41 / Chapter 3.1.2 --- Risk factors of cardiovascular disease --- p.42 / Chapter 3.1.3 --- Animal model --- p.43 / Chapter 3.2 --- Objectives --- p.44 / Chapter 3.3 --- Materials and Methods --- p.45 / Chapter 3.3.1 --- Animals --- p.46 / Chapter 3.3.2 --- Serum lipid and lipoprotein determinations --- p.46 / Chapter 3.3.3 --- Determination of cholesterol in the liver and adipose tissue --- p.46 / Chapter 3.3.4 --- Extraction of neutral and acidic sterols from fecal samples --- p.49 / Chapter 3.3.4.1 --- Determination of neutral sterols --- p.49 / Chapter 3.3.4.2 --- Determination of acidic sterols --- p.50 / Chapter 3.3.4.3 --- GLC analysis of neutral and acidic sterols --- p.51 / Chapter 3.3.5 --- Statistics --- p.51 / Chapter 3.4 --- Results --- p.54 / Chapter 3.4.1 --- Growth and food intake --- p.54 / Chapter 3.4.2 --- "Effects of SLP and SLEE supplementation on serum triacylglycerol (TG), total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C)" --- p.54 / Chapter 3.4.3 --- Effects ofSLP and SLEE supplementation on non-HDL-C and ratio of non-HDL-C to HDL-C --- p.55 / Chapter 3.4.4 --- Effects of SLP amd SLEE supplementations on concentration of hepatic cholesterol --- p.58 / Chapter 3.4.5 --- Effects of SLP and SLEE supplementations on perirenal adipose tissue cholesterol --- p.58 / Chapter 3.4.6 --- Effects of SLP and SLEE supplementations on fecal neutral and acidic sterols --- p.61 / Chapter 3.5 --- Discussion --- p.64 / Chapter Chapter 4 --- Effects of soy leaves and its flavonoid glycosides on haemolysis and on LDL oxidation / Chapter 4.1 --- Introduction --- p.67 / Chapter 4.1.1 --- Role of low density lipoprotein oxidation in the development of atherosclerosis --- p.68 / Chapter 4.1.2 --- LDL oxidation --- p.70 / Chapter 4.1.3 --- Thiobarbituric acid reactive substances (TBARS) as an index of LDL oxidation --- p.71 / Chapter 4.1.4 --- Antioxidant and LDL oxidation --- p.74 / Chapter 4.2 --- Objective --- p.75 / Chapter 4.3 --- Materials and methods --- p.76 / Chapter 4.3.1 --- Isolation of LDL from human serum --- p.76 / Chapter 4.3.2 --- LDL oxidation --- p.77 / Chapter 4.3.3 --- Determine the formation of thiobarbituric acid-reactive substances (TBARS) --- p.77 / Chapter 4.3.4 --- Assay for erythrocyte haemolysis --- p.78 / Chapter 4.3.5 --- Statistics --- p.79 / Chapter 4.4 --- Results --- p.80 / Chapter 4.4.1 --- Effects of three different soy leaves extracts and flavonoid glycosides on LDL oxidation --- p.80 / Chapter 4.4.2 --- Effects of three soy leaves extracts and flavonoid glycosides on erythrocyte haemolysis --- p.80 / Chapter 4.5 --- Discussion --- p.85 / Chapter Chapter 5 --- Relaxing effects of soy leaves and its flavonoids / Chapter 5.1 --- Introduction --- p.89 / Chapter 5.1.1 --- Smooth muscle contraction --- p.90 / Chapter 5.1.1.1 --- Sliding filament mechanism --- p.91 / Chapter 5.1.2 --- Intracellular mechanisms involved in the regulation of smooth muscle contraction --- p.92 / Chapter 5.1.2.1 --- Voltage-gated Ca2+ channels --- p.92 / Chapter 5.1.2.2 --- Protein kinase C (PKC) mediated smooth muscle contraction --- p.93 / Chapter 5.1.2.3 --- Thromboxane A2 receptor-mediated calcium channel --- p.94 / Chapter 5.2 --- Objectives --- p.96 / Chapter 5.3 --- Materials and methods --- p.97 / Chapter 5.3.1 --- Drugs preparation --- p.97 / Chapter 5.3.2 --- Vessel preparation --- p.97 / Chapter 5.3.3 --- Contraction experiments --- p.99 / Chapter 5.3.3.1 --- Relaxant responses of soy leaves butanol extract on the contraction induced by different constrictors --- p.99 / Chapter 5.3.3.2 --- Relaxant responses of soy leaves butanol extract on U46619 and PGF2a- induced contraction --- p.99 / Chapter 5.3.3.3 --- "Relaxant responses of genistein, genistin and the kaempferol glycosides on U46619-induced contraction" --- p.100 / Chapter 5.3.4 --- Statistics --- p.100 / Chapter 5.4 --- Results --- p.102 / Chapter 5.4.1 --- Effect of soy leaves butanol extract --- p.102 / Chapter 5.4.2 --- Role of endothelium in extract-induced relaxation --- p.102 / Chapter 5.4.3 --- Effect of the soy leaves butanol extract on contractile response to prostaglandins --- p.103 / Chapter 5.4.4 --- Effects of kaempferol glycosides and kaempferol --- p.111 / Chapter 5.4.5 --- Effects of genistein and genistin --- p.111 / Chapter 5.5 --- Discussion --- p.118 / Chapter Chapter 6 --- Effect of soy leaves on mammary tumor / Chapter 6.1 --- Introduction --- p.123 / Chapter 6.1.1 --- Carcinogenesis --- p.123 / Chapter 6.1.1.1 --- In itiation --- p.124 / Chapter 6.1.1.2 --- Promotion --- p.124 / Chapter 6.1.1.3 --- Progression --- p.125 / Chapter 6.2 --- Objective --- p.126 / Chapter 6.3 --- Materials and methods --- p.127 / Chapter 6.3.1 --- Animal --- p.127 / Chapter 6.3.2 --- Determination of estrus cycle --- p.128 / Chapter 6.3.3 --- Statistics --- p.129 / Chapter 6.4 --- Results --- p.131 / Chapter 6.4.1 --- Incident rate of tumor induction --- p.131 / Chapter 6.4.2 --- Number of tumor induced --- p.131 / Chapter 6.5 --- Discussion --- p.136 / Chapter Chapter 7 --- Conclusions --- p.136 / References --- p.140

Ho, Sai Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 144-164). / Abstracts in English and Chinese. / TABLE OF CONTENTS --- p.ii / ABSTRACT --- p.xii / 撮要 --- p.xv / LIST OF ABBREIVIATIONS --- p.xvi / LIST OF TABLES --- p.xvii / LIST OF FIGURES --- p.xix / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Overview of lectins --- p.1 / Chapter 1.1.1 --- History of lectins --- p.1 / Chapter 1.1.2 --- Definitions of lectins --- p.2 / Chapter 1.1.3 --- Classification and nomenclature of lectins based on structure --- p.2 / Chapter 1.1.4 --- Classification and nomenclature of lectins based on carbohydrate-bindingspecificity --- p.4 / Chapter 1.1.5 --- Structure of plant lectins --- p.4 / Chapter 1.1.6 --- Biological function of plant lectins --- p.5 / Chapter 1.1.6.1 --- Anti-viral activity of plant lectiins --- p.5 / Chapter 1.1.6.2 --- Lectins as plant defense proteins --- p.6 / Chapter 1.1.6.3 --- Insecticidal activity of plant lectins --- p.7 / Chapter 1.1.6.4 --- Anti-fungal activity of plant lectins --- p.7 / Chapter 1.1.6.5 --- Mitogenic activity of plant lectins --- p.7 / Chapter 1.1.6.6 --- Anti-tumor and anti-proliferative activity of plant lectins --- p.9 / Chapter 1.1.7 --- Background of legume lectins --- p.11 / Chapter 1.1.7.1 --- Structure of legume lectins --- p.11 / Chapter 1.1.7.2 --- Functions and activities of legume lectins --- p.12 / Chapter 1.2 --- Overview of serine protease inhibitors in plants --- p.14 / Chapter 1.2.1 --- Classification of serine protease inhibitor --- p.15 / Chapter 1.2.2 --- The main functions of plant serine protease inhibitors --- p.17 / Chapter 1.2.3 --- Commercial application of serine protease inhibirtors --- p.19 / Chapter 1.2.3.1 --- Medical application --- p.19 / Chapter 1.2.3.2 --- Transgenic application in agriculture --- p.22 / Chapter 1.3 --- Overview of Pathogenesis-related proteins in plants --- p.25 / Chapter 1.3.1 --- Overview of PR-5 family Thaumatin-like proteins (TLPs) --- p.27 / Chapter 1.3.1.1 --- Structural similarities among TLPs --- p.28 / Chapter 1.3.1.2 --- Antifungal activity of TLP --- p.31 / Chapter 1.3.2 --- Overview of Chinase-like proteins (CLPs) --- p.33 / Chapter 1.3.2.1 --- Classification of chitinase --- p.34 / Chapter 1.3.2.1.1 --- On the basis of amino acid sequence of glycosyl hydrolase --- p.34 / Chapter 1.3.2.1.2 --- On the basis of amino acid sequence of plant chitinase --- p.35 / Chapter 1.3.2.2 --- Antifungal activity of CLP --- p.36 / Chapter 1.3.3 --- Anti-freeze property of PR proteins --- p.38 / Chapter 1.3.4 --- Application of PR proteins in agriculture --- p.40 / Chapter 1.4 --- Rationale of the present study --- p.42 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials --- p.43 / Chapter 2.2 --- Preparation of crude extract --- p.44 / Chapter 2.2.1 --- Hokkaido large black soybean --- p.44 / Chapter 2.2.2 --- Emperor banana --- p.45 / Chapter 2.3 --- Purification --- p.45 / Chapter 2.4 --- Chromatography --- p.46 / Chapter 2.4.1 --- DEAE-cellulose chromatography --- p.46 / Chapter 2.4.2 --- Affi-gel Blue gel --- p.47 / Chapter 2.4.3 --- SP-Sepharse --- p.48 / Chapter 2.4.4 --- Mono Q HR 5/5 and Mono S HR 5/5 --- p.49 / Chapter 2.4.5 --- Superdex 75 and superdex 200 --- p.50 / Chapter 2.5 --- Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.50 / Chapter 2.6 --- Protein concentration determination --- p.54 / Chapter 2.7 --- Preparation of rabbit reticulocyte lysate --- p.54 / Chapter 2.8 --- Determination of N-terminal amino acid sequence --- p.56 / Chapter 2.9 --- Assay of inhibition of hemagglutinating activity by different carbohydrates --- p.56 / Chapter 2.10 --- Thermal stability determination assays --- p.57 / Chapter 2.10.1 --- Stability at various temperatures --- p.57 / Chapter 2.10.2 --- Stability at 100°C --- p.57 / Chapter 2.11 --- Assay of pH dependence of hemagglutinating activity --- p.58 / Chapter 2.12 --- Assay of ion dependence of hemagglutinating activity --- p.58 / Chapter 2.13 --- Assay of antifungal activity --- p.58 / Chapter 2.14 --- Assay of trypsin inhibitory activity --- p.60 / Chapter 2.15 --- Assay of antibacterial activity --- p.61 / Chapter 2.16 --- Assay for cytotoxic activity on cancer cell lines --- p.61 / Chapter 2.17 --- Assay for HIV-1 reverse transcriptase (RT) inhibitory activity --- p.62 / Chapter 2.18 --- Assay of mitogenic activity --- p.63 / Chapter Chapter 3 --- Purification and Characterization of Defense-Related Proteins from their Respective Sources / Chapter 3.1 --- Purification and Characterization of a Lectin from the Seeds of Hokkaido large black soybean / Chapter 3.1.1 --- Introduction --- p.65 / Chapter 3.1.2 --- Results --- p.66 / Chapter 3.1.3 --- Purification --- p.68 / Chapter 3.1.3.1 --- Affinity chromatography on Affi-gel Blue gel --- p.69 / Chapter 3.1.3.2 --- Anion-exchange chromatography on DEAE-cellulose --- p.70 / Chapter 3.1.3.3 --- Anion-exchange chromatography on Mono Q column --- p.71 / Chapter 3.1.3.4 --- Gel filtration on Superdex 200 column --- p.72 / Chapter 3.1.3.5 --- Hemagglutinating activity at each purification step --- p.73 / Chapter 3.1.4 --- Characterization of Lectin --- p.74 / Chapter 3.1.4.1 --- Molecular mass determination --- p.74 / Chapter 3.1.4.2 --- N-terminal amino acid sequencing --- p.76 / Chapter 3.1.4.3 --- Assay of inhibition of hemagglutinating activity by different carbohydrates --- p.77 / Chapter 3.1.4.4 --- Thermal stability --- p.78 / Chapter 3.1.4.5 --- Assay of pH dependence of hemagglutinating activity --- p.80 / Chapter 3.1.4.6 --- Assay of ion dependence of hemagglutinating activity --- p.81 / Chapter 3.1.4.7 --- Assay for HIV-1 reverse transcriptase (RT) inhibitory activity --- p.82 / Chapter 3.1.4.8 --- Assay of mitogenic activity --- p.83 / Chapter 3.1.4.9 --- Assay of antibacterial activity --- p.84 / Chapter 3.1.5 --- Discussion --- p.86 / Chapter 3.2 --- Purification and Characterization of a Trypsin inhibitor from the Seeds of Hokkaido large black soybean / Chapter 3.2.1 --- Introduction --- p.93 / Chapter 3.2.2 --- Results --- p.94 / Chapter 3.2.3 --- Purification --- p.95 / Chapter 3.2.3.1 --- Anion-exchange chromatography on Mono Q column --- p.96 / Chapter 3.2.3.2 --- Gel filtration on Superdex 75 column --- p.98 / Chapter 3.2.3.3 --- Trypsin inhibitory activity at each purification step --- p.99 / Chapter 3.2.4 --- Characterization of trypsin inhibitory --- p.100 / Chapter 3.2.4.1 --- Molecular mass determination --- p.100 / Chapter 3.2.4.2 --- N-terminal amino acid sequencing --- p.102 / Chapter 3.2.4.3 --- Assay for HIV-1 reverse transcriptase (RT) inhibitory activity --- p.103 / Chapter 3.2.4.4 --- Antiproliferative effect on MCF-7 and Hep G2 cells --- p.104 / Chapter 3.2.4.5 --- pH and thermal stability --- p.105 / Chapter 3.2.5 --- Discussion --- p.106 / Chapter 3.3 --- Purification and Characterization of a Thaumatin-like protein and Chitinase-like protein from Emperor Banana / Chapter 3.3.1 --- Introduction --- p.108 / Chapter 3.3.2 --- Results --- p.109 / Chapter 3.3.3 --- Purification --- p.111 / Chapter 3.3.3.1 --- Affinity chromatography on Affi-gel Blue gel --- p.112 / Chapter 3.3.3.2 --- Cation exchange chromatography on Mono S column --- p.113 / Chapter 3.3.3.3 --- Gel filtration on Superdex 75 column --- p.114 / Chapter 3.3.3.3.1 --- Fraction MS 2 --- p.114 / Chapter 3.3.3.3.2 --- Fraction MS 4 --- p.115 / Chapter 3.3.3.3.3 --- Fraction MS 5 --- p.118 / Chapter 3.3.4 --- Characterization of the thaumatin-like protein --- p.121 / Chapter 3.3.4.1 --- N-terminal amino acid sequence determination --- p.121 / Chapter 3.3.4.2 --- Assay for antifungal activity --- p.122 / Chapter 3.3.4.3 --- Thermal stability --- p.124 / Chapter 3.3.4.4 --- pH stability --- p.125 / Chapter 3.3.4.5 --- Resistance to trypsin digestion --- p.125 / Chapter 3.3.4.6 --- Anti-HIV-1 reverse transcriptase activity --- p.126 / Chapter 3.3.4.7 --- Discussion --- p.127 / Chapter 3.3.5 --- Characterization of the two chitinase-like protein --- p.131 / Chapter 3.3.5.1 --- N-terminal amino acid sequence determination --- p.131 / Chapter 3.3.5.1.1 --- Emperor banana MS2 CLP --- p.131 / Chapter 3.3.5.1.2 --- Emperor banana MS4 CLP --- p.132 / Chapter 3.3.5.2 --- Assay for antifungal activity --- p.133 / Chapter 3.3.5.3 --- Discussion --- p.136 / Chapter Chapter 4 --- general discussion --- p.138 / References --- p.144