Antitumor activities of tremella aurantialba polysaccharides.

Choi Pui-yu.Thesis (M.Phil.)--Chinese University of Hong Kong, 2002.Includes bibliographical references (leaves 113-123).Abstracts in English and Chinese.Abstract --- p.iAbstract (Chinese Version) --- p.iiiAcknowledgements --- p.vList of Abbreviations --- p.viTable of Contents --- p.viiiList of Tables --- p.xiiList of Figures --- p.xiiiChapter Chapter 1 --- Introduction --- p.1Chapter Chapter 2 --- Literature ReviewChapter 2.1 --- Mushroom Polysaccharides with Antitumor Activities --- p.7Chapter 2.1.1 --- Antitumor β-Glucans --- p.7Chapter 2.1.2 --- Antitumor Heteroglycans --- p.9Chapter 2.1.3 --- Antitumor Polysaccharide-Protein Complexes --- p.12Chapter 2.2 --- Antitumor Activities and Structural Characteristics of Mushroom Polysaccharides --- p.15Chapter 2.3 --- Antitumor Effects of Mushroom Polysaccharides In Vitro --- p.19Chapter 2.4 --- Antitumor Effects of Mushroom Polysaccharides In Vivo --- p.21Chapter 2.5 --- Immunomodulatory Activities --- p.24Chapter 2.6 --- Activation of Cytokines by Mushroom Polysaccharides --- p.28Chapter 2.7 --- Induction of Nitric Oxide Synthase by Mushroom Polysaccharides --- p.32Chapter 2.8 --- Tremella aurantialba --- p.34Chapter Chapter 3 --- Materials and Methods --- p.35Chapter 3.1 --- Extraction --- p.35Chapter 3.1.1 --- Extraction of Crude Tremella aurantialba Polysaccharide --- p.35Chapter 3.1.2 --- Fractionation --- p.38Chapter 3.1.3 --- Polysaccharide and Protein Content Determination --- p.38Chapter 3.1.3.1 --- Phenol-Sulfuric Assay --- p.39Chapter 3.1.3.2 --- Lowry-Folin Method --- p.39Chapter 3.1.4 --- Gas Chromatography (GC) --- p.40Chapter 3.1.5 --- Modified Carbazole Assay --- p.41Chapter 3.1.6 --- High Performance Liquid Chromatography (HPLC) --- p.42Chapter 3.2 --- In Vitro Studies --- p.43Chapter 3.2.1 --- Maintenance of Cell Lines --- p.43Chapter 3.2.2 --- Effect on Cancer Cell Lines --- p.43Chapter 3.2.2.1 --- Trypan Blue Exclusion Methods --- p.44Chapter 3.2.2.2 --- MTT Assay --- p.44Chapter 3.2.3 --- Effect on Normal Cell Line --- p.45Chapter 3.2.4 --- Coulter Counter --- p.46Chapter 3.3 --- In Vivo Studies --- p.47Chapter 3.3.1 --- Animals --- p.47Chapter 3.3.2 --- Maintenance of Sarcoma 180 Cell Line --- p.47Chapter 3.3.3 --- Effect of TAP Fractions on Sarcoma 18 Solid Tumor --- p.48Chapter 3.3.3.1 --- Injection of TAP Fractions 24 h After Transplantation --- p.48Chapter 3.3.3.2 --- Injection of TAP Fractions 72 h After Transplantation --- p.49Chapter 3.4 --- Effect of TAP Fractions on Modulating mRNA Expression of Cytokines and Nitric Oxide Synthase --- p.51Chapter 3.4.1 --- Treatment of Mice --- p.51Chapter 3.4.2 --- Isolation of Splenocytes and Peritoneal Exduate Cells --- p.51Chapter 3.4.3 --- Extraction of Total mRNA from Splenocyte and Peritoneal Exduate Cells --- p.52Chapter 3.4.4 --- Reverse Transcription-Polymerase Chain Reaction (RT-PCR) --- p.53Chapter 3.4.4.1 --- Reverse Transcription --- p.53Chapter 3.4.4.2 --- Polymerase Chain Reaction --- p.56Chapter 3.4.5 --- DNA Sequencing --- p.57Chapter 3.5 --- Statistical Analysis --- p.58Chapter Chapter 4 --- Results --- p.59Chapter 4.1 --- Isolation and Characterization of TAP Fractions --- p.59Chapter 4.1.1 --- Percentage Yield of TAP Fractions --- p.59Chapter 4.1.2 --- Polysaccharide and Protein Content of TAP Fractions --- p.59Chapter 4.1.3 --- Relative Monosaccharide Contents in TAP Fractions --- p.60Chapter 4.1.4 --- Results of HPLC --- p.60Chapter 4.2 --- Effects of TAP Fractions In Vitro --- p.69Chapter 4.2.1 --- Effects of TAP Fractions on Suspension Cancer Cell Lines --- p.69Chapter 4.2.2 --- Effects of TAP Fractions on Adhesion Cancer Cell Lines --- p.69Chapter 4.2.3 --- Effects of TAP Fractions on Normal Cell Line --- p.70Chapter 4.2.4 --- Effect of TAP 2 on HL-60 Cell Line as Evaluated by Coulter Counter --- p.70Chapter 4.3 --- Antitumor Effect of TAP Fractions In Vivo --- p.78Chapter 4.4 --- Effect of TAP Fractions on Modulating mRNA Expressions of Cytokines and Nitric Oxide Sythase (NOS) --- p.83Chapter 4.4.1 --- Results of RT-PCR --- p.83Chapter 4.4.2 --- Sequencing --- p.84Chapter Chapter 5 --- Discussion --- p.91Chapter 5.1 --- Characterization of TAP Fractions --- p.91Chapter 5.2 --- Antitumor Effects of TAP Fractions In Vitro --- p.96Chapter 5.3 --- Furhter Study --- p.109Chapter Chapter 6 --- Conclusion --- p.111References --- p.11