by Fu Sai-chuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 102-111). / Acknowledgements / List of Abbreviation / Abstract / Prologue --- p.1 / Chapter / Chapter 1. --- Introduction --- p.2 / Chapter 1.1 --- Historical background of Ling Zhi (靈芝） --- p.4 / Chapter 1.2 --- Biological description of Ling Zhi (靈芝） --- p.6 / Chapter 1.3 --- Chemical composition of Ganoderma lucidum --- p.6 / Chapter 1.3.1 --- The triterpenoids --- p.7 / Chapter 1.3.2 --- Matrix polysaccharides --- p.8 / Chapter 1.4 --- Medicinal properties and biological activities --- p.9 / Chapter 1.4.1 --- Medicinal properties --- p.9 / Chapter 1.4.2 --- Biological activities --- p.10 / Chapter 1.4.2.1 --- Anti-tumour activity --- p.10 / Chapter 1.4.2.2 --- Enhancement of protein and nucleic acid synthesis --- p.11 / Chapter 1.4.2.3 --- Effects on nervous system --- p.11 / Chapter 1.4.2.4 --- Effects on respiratory system --- p.12 / Chapter 1.4.2.5 --- Effects on cardiovascular system --- p.12 / Chapter 1.4.2.6 --- Effects on immune system --- p.14 / Chapter 1.4.2.7 --- Hepatoprotection and detoxicant actions --- p.14 / Chapter 1.4.3 --- Summary --- p.15 / Chapter 1.5 --- The relationship between antioxidant defense/immune system and disease --- p.15 / Chapter 2. --- Materials and Methods --- p.20 / Chapter 2.1 --- Cell culture reagents / Chapter 2.1.1 --- Preparation of RPMI-1640 medium --- p.20 / Chapter 2.1.2 --- Preparation of fetal calf serum (FCS) --- p.20 / Chapter 2.1.3 --- Preparation of phosphate-buffered saline (PBS) --- p.20 / Chapter 2.1.4 --- Mitogens and test samples --- p.21 / Chapter 2.1.5 --- Trypan blue exclusion test --- p.21 / Chapter 2.1.6 --- Liquid scintillation counting --- p.21 / Chapter 2.2 --- Animal care --- p.22 / Chapter 2.3 --- Fractionation of Ganoderma lucidum extracts --- p.22 / Chapter 2.4 --- Carbon tetrachloride (CC14)- induced hepatotoxicity in mice --- p.23 / Chapter 2.4.1 --- Pretreatment scheme and CCl4 treatment --- p.23 / Chapter 2.4.2 --- Preparation of liver homogenates --- p.23 / Chapter 2.4.3 --- Biochemical assays for assessing hepatotoxicity --- p.25 / Chapter 2.4.3.1 --- Determination of plasma alanine aminotransferase (ALT) --- p.25 / Chapter 2.4.3.2 --- Determination of tissue glutathione (GSH) content --- p.25 / Chapter 2.4.3.3 --- Depletion of tissue GSH in liver homogenates by tert- butylhydroperoxide (tBHP) --- p.26 / Chapter 2.4.3.4 --- Determination of tissue malondialdehyde (MDA) content --- p.26 / Chapter 2.5 --- Preparation of murine hepatocytes --- p.27 / Chapter 2.5.1 --- Liver perfusion --- p.27 / Chapter 2.5.2 --- Isolation of murine hepatocytes --- p.29 / Chapter 2.5.3 --- Incubation protocol --- p.29 / Chapter 2.5.4 --- Determination of cellular glutathione by HPLC method --- p.30 / Chapter 2.6 --- Preparation of splenocytes and lymphocytes --- p.31 / Chapter 2.6.1 --- Isolation of murine splenocytes --- p.31 / Chapter 2.6.2 --- Mitogenic assay by measuring 3H-thymidine uptake --- p.31 / Chapter 2.6.3 --- B cell- and T cell- enrichment in splenocyte preparation --- p.32 / Chapter 2.6.4 --- Isolation of human peripheral blood lymphocytes (HPBL) from cord blood --- p.34 / Chapter 2.7 --- Chromatographic methods --- p.34 / Chapter 2.7.1 --- Fast Protein Liquid Chromatography (FPLC) with Superose 12 column --- p.34 / Chapter 2.7.2 --- Removal of lipopolysaccharide (LPS) with Detoxi Gel --- p.35 / Chapter 2.7.3 --- Desalting by Sephadex G-25 --- p.35 / Chapter 2.7.4 --- Determination of FDNB-conjugated GSH and GSSG by HPLC reverse phase chromatography --- p.36 / Chapter 2.8 --- Measurement of in vivo antibody production --- p.39 / Chapter 2.8.1 --- Immunization and pretreatment scheme --- p.39 / Chapter 2.8.2 --- Direct plaque assay --- p.39 / Chapter 2.9 --- Biochemical analysis --- p.39 / Chapter 2.9.1 --- Determination of protein content --- p.40 / Chapter 2.9.2 --- Determination of hexose content --- p.40 / Chapter 2.9.3 --- Determination of uronic acid content --- p.40 / Chapter 2.9.4 --- Determination of sulphate content --- p.41 / Chapter 2.9.5 --- Determination of hexosamine content --- p.42 / Chapter 2.10 --- Statistical analysis --- p.42 / Chapter 3. --- Results --- p.44 / Chapter 3.1 --- Extraction and fractionation of Ganoderma lucidum --- p.44 / Chapter 3.2 --- Hepatoprotective effect of Ganoderma lucidum fractions aganist CCl4-induced hepatotoxicity --- p.47 / Chapter 3.2.1 --- Hepatoprotection of Ganoderma lucidum fractions --- p.47 / Chapter 3.2.2 --- Effect of Ganoderma lucidum fraction pretreatment on hepatic malondialdehyde (MDA) level in CCl4-treated mice --- p.47 / Chapter 3.2.3 --- Effect of Ganoderma lucidum fraction pretreatment on hepatic glutathione (GSH) level in CCl4-treated mice --- p.51 / Chapter 3.3 --- Effect of GL2 on isolated hepatocytes --- p.51 / Chapter 3.3.1 --- Protection against phorone-induced hepatocytotoxicity --- p.51 / Chapter 3.3.2 --- Protection against menadione-induced hepatocytotoxicity --- p.55 / Chapter 3.4 --- Immunomodulatory effect of Ganoderma lucidum fractions --- p.55 / Chapter 3.4.1 --- Discovery of a mitogenic principle from Ganoderma lucidum fractions --- p.55 / Chapter 3.4.2 --- B cell-specific mitogenic activity --- p.58 / Chapter 3.4.3 --- Mitogenic activity in Ganoderma lucidum distinguishable from possible LPS contamination --- p.65 / Chapter 3.4.3.1 --- The use of polymyxin B sulphate --- p.65 / Chapter 3.4.3.2 --- The use of Detoxi Gel --- p.69 / Chapter 3.4.3.3 --- The combined use of polymyxin B sulphate and Detoxi Gel --- p.69 / Chapter 3.5 --- Enhancement of humoral response to sheep red blood cells (SRBC) by the mitogenic GL2 fraction --- p.72 / Chapter 3.6 --- Physical-chemical characterization of GL2 fraction --- p.76 / Chapter 3.6.1 --- Estimation of molecular size by FPLC size exclusion chromatography with Superose 12 column --- p.76 / Chapter 3.6.2 --- Sugar composition of GL2 fraction --- p.76 / Chapter 3.6.3 --- Comparison of elution profiles in Superose12 column with respect to mitogenic activity and sugar composition --- p.80 / Chapter 4. --- Discussions --- p.83 / Chapter 4.1 --- Enhancement of antioxidant status by Ganoderma lucidum fractions --- p.83 / Chapter 4.2 --- The significance of enhancement of GSH status --- p.89 / Chapter 4.3 --- Immunomodulatory actions of Ganoderma lucidum --- p.95 / Conclusions --- p.99 / Appendix --- p.100 / Reference --- p.102

Leung Yiu Kwong, Michael. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 155-161). / Acknowledgements --- p.i / Abbreviations --- p.ii / Aim and scope of this dissertation --- p.v / Abstract --- p.vi / Table of contents --- p.viii / Introduction --- p.1 / Chapter 1.1 --- Introduction --- p.2 / Chapter 1.2 --- Tumor Biology --- p.3 / Chapter 1.3 --- The Defence Mechanisms --- p.4 / Chapter 1.3.1 --- Non-specific defence mechanisms --- p.5 / Chapter 1.3.2 --- Specific defence mechanisms --- p.6 / Chapter 1.4 --- Effector Mechanisms in Anti-tumor Immunity --- p.7 / Chapter 1.4.1 --- B-cell --- p.8 / Chapter 1.4.2 --- "Natural killer (NK) cells (Non-T, Non-B)" --- p.8 / Chapter 1.4.3 --- Macrophages --- p.9 / Chapter 1.4.4 --- Cytolytic T-lymphocytes (CTLs) --- p.10 / Chapter 1.5 --- Cancer Treatment --- p.10 / Chapter 1.5.1 --- Surgery --- p.10 / Chapter 1.5.2 --- Radiotherapy --- p.12 / Chapter 1.5.3 --- Drug therapy --- p.12 / Chapter 1.5.4 --- Gene therapy --- p.13 / Chapter 1.5.5 --- lmmunotherapy --- p.13 / Chapter 1.6 --- Non-cytotoxic Antitumor Polysaccharides of Fungi --- p.14 / Chapter 1.6.1 --- Yeast polysaccharides --- p.14 / Chapter 1.6.2 --- Lichen polysaccharides --- p.15 / Chapter 1.6.3 --- Fungal polysaccharides --- p.18 / Chapter 1.7 --- Fungi and their Polysaccharides --- p.20 / Chapter 1.7.1 --- Reserve carbohydrates --- p.20 / Chapter 1.7.2 --- Structural polysaccharides --- p.21 / Chapter 1.8 --- The Architecture of the Fungal Cell Wall --- p.22 / Materials and Methods --- p.26 / Chapter 2.1 --- Materials --- p.27 / Chapter 2.1.1 --- Animals --- p.27 / Chapter 2.1.2 --- Mushrooms --- p.27 / Chapter 2.1.3 --- "Buffers, culture media and chemicals" --- p.27 / Chapter 2.1.4 --- Cell lines --- p.34 / Chapter 2.2 --- Methods --- p.35 / Chapter 2.2.1 --- Screening of β-(l→3)-D-glucan --- p.35 / Chapter 2.2.2 --- Extraction and Fractionation of Flammulina velutipes --- p.35 / Chapter 2.2.3 --- Characterisation of Flammulina velutipes --- p.38 / Chapter 2.2.3.1 --- The determination of carbohydrate content of F.V fractions --- p.38 / Chapter 2.2.3.2 --- The determination of protein content of F.V. fractions --- p.39 / Chapter 2.2.3.3 --- The determination of uronic acid content of F.V fractions --- p.39 / Chapter 2.2.3.4 --- The determination of component sugar units of F.V fractions --- p.39 / Chapter 2.2.3.5 --- Periodate uptake of F.V. fractions --- p.40 / Chapter 2.2.3.6 --- Limulus amebocyte lysate (LAL) coagulation assay --- p.40 / Chapter 2.2.3.7 --- The digestion of F.V. fractions with laminarinase --- p.41 / Chapter 2.2.3.8 --- The Secondary and tertiary structure determination of FH and SFA1 --- p.42 / Chapter 2.2.3.9 --- Molecular weight estimation of FH and SFA1 --- p.43 / Chapter 2.2.3.10 --- Vascular dilation and hemorrhage (VDH) activity of F.V. fractions / Chapter 2.2.4 --- Isolation and preparation of cells --- p.43 / Chapter 2.2.4.1 --- Bone marrow cell --- p.43 / Chapter 2.2.4.2 --- Peritoneal exudate cell (PEC) --- p.44 / Chapter 2.2.4.3 --- Splenocytes --- p.44 / Chapter 2.2.4.4 --- Depleting mouse T-cells by anti-mouse T-cell antigen antibody plus complement treatment --- p.45 / Chapter 2.2.4.5 --- Depleting mouse B-cells by Cedarlane column kit --- p.45 / Chapter 2.2.5 --- Assays for the cytotoxicity of Flammulina velutipes --- p.45 / Chapter 2.2.5.1 --- Brine shrimp assay --- p.45 / Chapter 2.2.5.2 --- In vitro cytotoxicity of FH and SFA1 on bone marrow cells of female BALB/c mice --- p.46 / Chapter 2.2.5.3 --- In vivo cytotoxicity of FH and SFA1 on female BALB/c mice --- p.47 / Chapter 2.2.6 --- "Assays for the immunomodulatory activities of Flamm""lina velutipes" --- p.47 / Chapter 2.2.6.1 --- In vitro mitogenic activities of FH and SFA1 on murine lymphocytes --- p.47 / Chapter 2.2.6.2 --- In vitro mitogenic activities of FH and SFA1 with PMB on murine lymphocytes --- p.48 / Chapter 2.2.6.3 --- In vitro mitogenic activities of FH and on T-cell depleted murine lymphocytes --- p.48 / Chapter 2.2.6.4 --- In vitro mitogenic activities of FH on B-cell depleted murine lymphocytes --- p.49 / Chapter 2.2.6.5 --- In vitro co-mitogenic activitiy of FH and SFA1 on murine lymphocytes --- p.49 / Chapter 2.2.6.6 --- In vitro mitogenic activities of FH and SFA1 on murine bone marrow cells --- p.50 / Chapter 2.2.6.7 --- In vivo mitogenic activities of FH and SFA1 on murine lymphocytes --- p.50 / Chapter 2.2.6.8 --- Effect of FH and SFA1 on the enhancement of first antibody production of SRBC immunised mice --- p.51 / Chapter 2.2.6.9 --- Effect of FM and SFA1 on the in vitro phagocytic activity of murine macrophage --- p.51 / Chapter 2.2.6.10 --- Effect of FM and SFA1 on the in vivo phagocytic activity of murine macrophage --- p.51 / Chapter 2.2.6.11 --- In vivo migration of macrophage in FH- and SFAl-treated mice --- p.53 / Chapter 2.2.6.12 --- Effect of FH and SFA1 on the enhancement of murine PEC cytostatic activity --- p.53 / Chapter 2.2.6.13 --- Effect of FH and SFA1 on the Fc receptor expression of peritoneal exudate cells --- p.54 / Chapter 2.2.6.14 --- Effect of FH and SFA1 on murine serum cytokine level --- p.55 / Chapter 2.2.6.15 --- Effect of FH and SFA1 on murine serum TNF level --- p.55 / Chapter 2.2.6.16 --- Effect of FH and SFA1 on the augmentation of SRBC lysing ability of murine serum --- p.56 / Chapter 2.2.7 --- Assays for the anti-tumor activities of Flammulina velutipes --- p.57 / Chapter 2.2.7.1 --- In vitro anti-tumor activity of FH and SFA1 --- p.57 / Chapter 2.2.7.2 --- Effect of FH and SFA1 on the growth of murine transplantable tumor invivo --- p.58 / Chapter 2.2.8 --- Statistical analysis --- p.59 / "Screening, Purification, Fractionation and Characterisation of β-(l→3)-D-glucan(s) from Flammulina velutipes" --- p.60 / Introduction --- p.61 / Results --- p.62 / Chapter 3.1 --- Screening of β-(l→3)-D-Glucan --- p.62 / Chapter 3.2 --- Extraction and Fractionation of Flammulina velutipes --- p.62 / Chapter 3.3 --- The Determination of Carbohydrate Content of F.V. Fractions --- p.65 / Chapter 3.4 --- The Determination of Protein Content of F.V. Fractions --- p.65 / Chapter 3.5 --- The Determination of Uronic Acid Content of F.V. Fractions --- p.69 / Chapter 3.6 --- The Determination of Component Sugar Units of F.V. Fractions --- p.69 / Chapter 3.7 --- Periodate Uptake of F.V. Fractions --- p.69 / Chapter 3.8 --- Limulus Amebocyte Lysate (LAL) Coagulation Assay --- p.73 / Chapter 3.9 --- The Digestion of F.V. Fractions with Laminarinase --- p.73 / Chapter 3.10 --- The Secondary and tertiary Structure Determination of FH and SFA1 --- p.80 / Chapter 3.11 --- Molecular Weight Estimation of FH and SFA1 --- p.82 / Chapter 3.12 --- "Vascular Dilation and Hemorrhage (VDH) Activity of FH, SFA1 and lFA1" --- p.82 / Discussion --- p.90 / The Toxicity of Flammulina velutipes --- p.96 / Introduction --- p.97 / Results --- p.97 / Chapter 4.1 --- Lack of Cytotoxicity of Flammulina velutipes to Brine Shrimp --- p.97 / Chapter 4.2 --- Lack of Cytotoxicity of Flammulina velutipes to Murine Bone Marrow Cells --- p.99 / Chapter 4.3 --- Lack of Cytotoxicity of Flammulina velutipes to Mouse --- p.99 / Discussion --- p.102 / The Immunomodulatory Activities of Flammulina velutipes --- p.103 / Introduction --- p.104 / Results --- p.105 / Chapter 5.1 --- Effect of Flammulina velutipes on Murine Lymphocytes --- p.105 / Chapter 5.2 --- Effect of Flammulina velutipes on Murine Macrophage --- p.115 / Chapter 5.3 --- Effect of Flammulina velutipes on Murine Serum Cytokine and Complement Level --- p.125 / Discussion --- p.133 / The Anti-tumor Activities of Flammulina velutipes --- p.136 / Introduction --- p.137 / Results --- p.137 / Chapter 6.1 --- In Vitro Anti-Tumor Activity of FH and SFA1 --- p.137 / Chapter 6.2 --- Effect of FH and SFAI on the Growth of Murine TransplantableTumors --- p.138 / Discussion --- p.145 / General Discussion --- p.146 / General Discussion and Future Perspectives --- p.147 / References --- p.154