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
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_318227 |
| Date | January 1994 |
| Contributors | Fu, Sai-chuen., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Publisher | Chinese University of Hong Kong |
| Source Sets | The Chinese University of Hong Kong |
| Language | English |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, [8], 111 leaves : ill. (some mounted 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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