by Wang He-Xiang. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 161-179). / ACKNOWLEDGEMENTS --- p.i / ABSTRACT --- p.ii / LIST OF CONTENTS --- p.v / LIST OF TABLES --- p.xi / LIST OF FIGURES --- p.xii / LIST OF ABBREVIATIONS --- p.xvi / Chapter CHAPTER 1. --- General Introduction --- p.1 / Chapter CHAPTER 2. --- Literature Review --- p.5 / Chapter 2.1. --- Lectins --- p.5 / Chapter 2.1.1. --- Aspects of lectins --- p.5 / Chapter 2.1.2. --- Isolation and purification of lectins --- p.8 / Chapter 2.1.3. --- Characteristics of lectins --- p.9 / Chapter 2.1.4. --- Effects of lectins on biological activities --- p.10 / Chapter 2.1.4.1. --- The role of lectins in plant defence --- p.11 / Chapter 2.1.4.2. --- The specificity of some legume lectins --- p.13 / Chapter 2.1.4.3. --- Some properties of animal lectins --- p.14 / Chapter 2.1.4.4. --- Hypotensive activity of the lectins --- p.18 / Chapter 2.1.4.5. --- Lectins in immunology --- p.20 / Chapter 2.2. --- Mushroom Lectins and Polysaccharides --- p.24 / Chapter 2.2.1. --- General aspects of mushroom lectins and polysaccharides --- p.24 / Chapter 2.2.2. --- Mushroom lectins --- p.25 / Chapter 2.2.2.1. --- Hericium erinaceum lectin --- p.26 / Chapter 2.2.2.2. --- Lactarius deterrimus lectin --- p.26 / Chapter 2.2.2.3. --- Laetiporus sulfureus lectin --- p.27 / Chapter 2.2.2.4. --- Grifola frondosa lectin --- p.28 / Chapter 2.2.2.5. --- Volvariella volvacea lectin --- p.28 / Chapter 2.2.2.6. --- Flammulina veltipes lectin --- p.29 / Chapter 2.2.2.7. --- Ischnoderma resinosum agglutinin --- p.31 / Chapter 2.2.2.8. --- Lectins from Agaricus spp --- p.31 / Chapter 2.2.3. --- Mushroom polysaccharides --- p.34 / Chapter 2.2.3.1. --- Lentinan --- p.35 / Chapter 2.2.3.2. --- "PSK (trade name, Krestin)" --- p.35 / Chapter 2.2.3.3. --- PSP (Polysaccharopeptide) --- p.37 / Chapter 2.2.3.4. --- PSPC (polysaccharide-peptide complex) --- p.38 / Chapter CHAPTER 3. --- Isolation and Characterization of Two Distinct Lectins from the Cultured Mycelium of the Edible Mushroom Tricholoma mongolicum --- p.44 / Chapter 3.1. --- Introduction --- p.44 / Chapter 3.2. --- Materials and Methods --- p.45 / Chapter 3.2.1. --- Strain and culture condition --- p.45 / Chapter 3.2.2. --- Extraction --- p.46 / Chapter 3.2.3. --- Purification --- p.46 / Chapter 3.2.4. --- Hemagglutination activity --- p.47 / Chapter 3.2.5. --- Test of hemagglutination inhibition by various carbohydrates --- p.47 / Chapter 3.2.6. --- MW estimation by gel filtration and SDS- PAGE --- p.48 / Chapter 3.2.7. --- Glycoprotein staining with PAS reagent --- p.49 / Chapter 3.2.8. --- Carbohydrate content --- p.49 / Chapter 3.2.9. --- Thermal stability --- p.49 / Chapter 3.2.10. --- pH stability --- p.49 / Chapter 3.2.11. --- Effect of cations --- p.50 / Chapter 3.2.12. --- Amino acid analysis --- p.50 / Chapter 3.2.13. --- Antiproliferative activity of lectins --- p.50 / Chapter 3.2.14. --- Statistics --- p.51 / Chapter 3.3. --- Results --- p.51 / Chapter 3.3.1. --- Extraction and purification --- p.51 / Chapter 3.3.2. --- General characteristics of lectins --- p.52 / Chapter 3.3.3. --- Antiproliferative activity of lectins --- p.54 / Chapter 3.4. --- Discussion --- p.55 / Chapter 3.5. --- Summary --- p.58 / Chapter CHAPTER 4. --- The Immunomodulatory and Antitumor Activities of Lectins from the Mushroom Tricholoma mongolicum --- p.79 / Chapter 4.1. --- Introduction --- p.79 / Chapter 4.2. --- Materials and Methods --- p.81 / Chapter 4.2.1. --- Lectins --- p.81 / Chapter 4.2.2. --- Animals --- p.81 / Chapter 4.2.3. --- Assay for antitumor activity --- p.81 / Chapter 4.2.4. --- Assessment of tumor growth and host survival after lectin treatment --- p.82 / Chapter 4.2.5. --- Mitogenic activity of lectins --- p.82 / Chapter 4.2.6. --- Production of nitrite ions in response to lectin treatment --- p.83 / Chapter 4.2.7. --- Preparation of concanavalin A-stimulated lymphokines --- p.84 / Chapter 4.2.8. --- Assay for macrophage activating factor --- p.85 / Chapter 4.2.9. --- Production of tumor necrosis factor (TNF) --- p.86 / Chapter 4.2.10. --- Bioassay for tumor necrosis factor --- p.86 / Chapter 4.2.11. --- Statistics --- p.87 / Chapter 4.3. --- Results --- p.87 / Chapter 4.3.1. --- Antitumor activity --- p.87 / Chapter 4.3.2. --- Assessment of tumor growth and host survival --- p.87 / Chapter 4.3.3. --- Mitogenic activity --- p.88 / Chapter 4.3.4. --- Production of nitrite ions --- p.89 / Chapter 4.3.5. --- Production of macrophage activating factor --- p.89 / Chapter 4.3.6. --- Tumor necrosis factor assay --- p.90 / Chapter 4.4. --- Discussion --- p.90 / Chapter 4.5. --- Summary --- p.94 / Chapter CHAPTER 5. --- Hypotensive and Vasorelaxing Activities of a Lectin (TML-1) from the Edible Mushroom Tricholoma mongolicum --- p.109 / Chapter 5.1. --- Introduction --- p.109 / Chapter 5.2. --- Materials and Methods --- p.111 / Chapter 5.2.1. --- Animals --- p.111 / Chapter 5.2.2. --- In vivo blood pressure measurement in rats --- p.112 / Chapter 5.2.3. --- Study employing blockade of autonomic ganglion transmission --- p.113 / Chapter 5.2.4. --- Study employing alpha-adrenergic blockade --- p.113 / Chapter 5.2.5. --- Study employing beta-adrenergic blockade --- p.114 / Chapter 5.2.6. --- Study employing cholinergic blockade --- p.114 / Chapter 5.2.7. --- Study employing histaminergic blockade --- p.114 / Chapter 5.2.8. --- Study employing inhibitor of the renin- angiotensin system --- p.115 / Chapter 5.2.9. --- Preparation of right atrium for in vitro studies --- p.115 / Chapter 5.2.10. --- Preparation of aorta for in vitro studies --- p.116 / Chapter 5.2.11. --- Adenosine receptor binding assays --- p.116 / Chapter 5.2.12. --- Effect of methylene blue on the hypotensive activity of TML-1 --- p.118 / Chapter 5.2.13. --- Statistics --- p.118 / Chapter 5.3. --- Results --- p.118 / Chapter 5.3.1. --- Blood pressure changes in vivo --- p.118 / Chapter 5.3.2. --- Pharmacological studies using receptor antagonists --- p.119 / Chapter 5.3.3. --- Adenosine receptor binding assay --- p.119 / Chapter 5.3.4. --- Effects on the right atrium in vitro --- p.120 / Chapter 5.3.5. --- Effect of TML-1 on vascular relaxation --- p.120 / Chapter 5.3.6. --- Effect of methylene blue on the hypotensive activity of TML-1 --- p.120 / Chapter 5.4. --- Discussion --- p.120 / Chapter 5.5. --- Summary --- p.123 / Chapter CHAPTER 6. --- A Polysaccharide-Peptide Complex with Immunoenhancing and Antitumor Activities from Cultured Mycelia of the Mushroom Tricholoma mongolicum --- p.134 / Chapter 6.1. --- Introduction --- p.134 / Chapter 6.2. --- Materials and Methods --- p.135 / Chapter 6.2.1. --- Extraction --- p.135 / Chapter 6.2.2. --- Purification --- p.135 / Chapter 6.2.3. --- PSP for purpose of comparison --- p.136 / Chapter 6.2.4. --- Polysaccharide and protein contents --- p.136 / Chapter 6.2.5. --- MW determination of F1 using gel filtration --- p.136 / Chapter 6.2.6. --- Animals --- p.136 / Chapter 6.2.7. --- Antiproliferative activity assay --- p.137 / Chapter 6.2.8. --- Mitogenic activity --- p.137 / Chapter 6.2.9. --- Production of nitrite ions --- p.138 / Chapter 6.2.10. --- Macrophage activating factor assay --- p.138 / Chapter 6.2.11. --- Antitumor activity assay --- p.139 / Chapter 6.2.12. --- Statistics --- p.139 / Chapter 6.3. --- Results --- p.140 / Chapter 6.3.1. --- Purification of polysaccharide-peptide complex --- p.140 / Chapter 6.3.2. --- Antiproliferative activity --- p.140 / Chapter 6.3.3. --- Mitogenic activity in vitro --- p.140 / Chapter 6.3.4. --- Molecular weight of Fl --- p.141 / Chapter 6.3.5. --- Mitogenic activity in vivo --- p.141 / Chapter 6.3.6. --- Production of nitrite ions --- p.141 / Chapter 6.3.7. --- Production of macrophage activating factor --- p.141 / Chapter 6.3.8. --- Antitumor activity in vivo --- p.142 / Chapter 6.4. --- Discussion --- p.142 / Chapter 6.5. --- Summary --- p.144 / GENERAL DISCUSSION --- p.155 / CONCLUSIONS --- p.158 / REFERENCES

Athymic nude mice were employed as the in vivo model to study the detailed mechanism of how the three sclerotial polysaccharides act to inhibit the growth of human xenografted tumors in vivo. Using immunohistochemical staining, it was found that the presence of F4/80 + macrophages was related to the reduction of tumor size of the HL-60 xenograft. mRNA extracted from the spleens were reverse-transcribed to cDNA and detected by real-time PCR so that a variety of genes related to the toll-like receptors being up-regulated or down-regulated due to the injection of mushroom sclerotial polysaccharides were determined. Combining the results from dectin-1 regulation, it was concluded that both hot water-soluble sclerotial polysaccharides, PTRW and PRW, having a structure of polysaccharide-protein complexes were responsible for activating and thus binding to CR3 or toll-like receptors while PRSon with structure of pure beta-glucan was responsible for activating the expression of dectin-1 receptor, which led to the subsequent activation of host immune system in immunopotentiation and antitumor activities. / In the future, further investigation of the detailed structure of mushroom sclerotial polysaccharides is required to explain the immunomodulatory mechanism so that the effective dosage for immunomodulation as well as antitumor effects can be determined. Furthermore, phage display can be applied to find out any novel glucan receptors specific to the mushroom sclerotial polysaccharides. / In vitro antitumor study indicated that PTRW had a significant (p<0.05) inhibitory effect (>40%) on the human monocytic leukemic cells (THP-1) in addition to HL-60 and K562 cells. In vitro immunomodulatory study showed that both PRW and PRSon had significant proliferative effects (p<0.05) on human normal spleen monocyte/macrophage cell, MD. Moreover, PRSon was shown to have a significant increase (p<0.05) in the growth of human natural killer cells, NK-92M1; however, PTRW showed a significant inhibition (p<0.05) on this cell line. / Mushroom sclerotia have a rich source of polysaccharides when compared with fruit bodies. It was previously reported that the polysaccharides from novel mushroom sclerotia, namely, Pleurotus tuber-regium and Polyporus rhinocerus, had potent in vitro and in vivo antitumor effects. In this project, hot water-soluble sclerotial polysaccharides of Pleurotus tuber-regium (PTRW), hot water-soluble and sonication-assisted cold alkali-soluble sclerotial polysaccharides of Polyporus rhinocerus (PRW and PRSon, respectively) were chosen for investigation of their in vivo and in vitro immunomodulatory effects. / Polysaccharides have long been proposed to exert their antitumor and thus immunomodulating functions through glucan receptors and among the four being discovered, Dectin-1 has drawn most attention recently. In the in vivo study, PRSon showed an increase in the expression of Dectin-1 on mice spleen MNCs while PTRW showed an increase in the expression of the previously widely-reported complement receptor (CR3). There was also an increase of Dectin-1 expression on PEC in the mice injected with PRSon. In the in vitro study, the three mushroom sclerotial polysaccharides were incubated with NK-92M1, MD and THP-1 cells. There was a significant increase (p<0.05) of Dectin-1 expression on NK-92MI cells incubated with PTRW. On the other hand, PTRW caused a significant decrease ( p<0.05) of Dectin-1 expression while PRSon showed a significant increase (p<0.05) on THP-1 cells. The cytokine profile of extra-cellular media indicated that the inhibition of THP-1 cells by PTRW should be related to the innate immunity. In the in vitro study, human primary immune cells, CD56+ NK cells were used to incubate with sclerotial polysaccharides and there was a significant stimulation (p<0.05) of their growth when compared with the control. / The in vivo immunomodulatory study was carried out by injecting the abovementioned sclerotial polysaccharides intraperitoneal to 7-8 weeks old healthy male BALB/c mice. The spleens excised from groups injected with PTRW and PRW were found to have significant increase of weight ( p<0.001). Flow cytometric analysis revealed that the NK cell population in spleen mononuclear cells (MNCs) of mice injected with PRW and PRSon was increased when compared with the control. In addition, ail three sclerotial polysaccharides showed a large increase of T helper cell population as well as CD4+/CD8+ ratio in spleen MNCs. On the other hand, the macrophage population in peritoneal exudates cells (PEC) was found to be increased in the groups of mice injected with PTRW and PRW. / Lai, Kin Ming Connie. / Adviser: Cheung Chi Keung. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3412. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 120-137). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

by Liu Fang. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 159-178). / ACKNOWLEGEMENTS --- p.i / ABSTRACT --- p.ii / TABLE OF CONTENTS --- p.v / LIST OF TABLES --- p.ix / LIST OF FIGURES --- p.xi / ABBREVIATIONS --- p.xiv / Chapter Chapter 1. --- General Introduction --- p.1 / Chapter Chapter 2. --- Literature Review --- p.7 / Chapter 2.1. --- Biologically active polysaccharides --- p.7 / Chapter 2.2. --- Antitumor activities of polysaccharides --- p.11 / Chapter 2.2.1. --- In vivo studies --- p.11 / Chapter 2.2.2. --- In vitro studies --- p.15 / Chapter 2.3. --- Antitumor mechanisms of polysaccharides --- p.17 / Chapter 2.4. --- Structure and antitumor activities of polysaccharides --- p.25 / Chapter 2.4.1. --- The effect of molecular mass --- p.26 / Chapter 2.4.2. --- The impact of branching configuration --- p.21 / Chapter 2.4.3. --- The relationship between antitumor activity and conformation --- p.28 / Chapter 2.4.4. --- Improvement of antitumor activity by chemical modification --- p.29 / Chapter 2.5. --- Other biological activities --- p.30 / Chapter 2.5.1. --- Antiviral activity --- p.30 / Chapter 2.5.2. --- Antimicrobial activity --- p.31 / Chapter 2.5.3. --- Free radical scavenging activity --- p.32 / Chapter 2.5.4. --- Hepatic protective effect --- p.32 / Chapter Chapter 3. --- Isolation and Characterization of a Polysaccharide-Protein Complex (PSPC) from Tricholoma lobayense --- p.34 / Chapter 3.1. --- Introduction --- p.34 / Chapter 3.2. --- Materials and methods --- p.36 / Chapter 3.2.1. --- Strain --- p.36 / Chapter 3.2.2. --- Culture conditions --- p.36 / Chapter 3.2.3. --- Extraction of T. lobayense --- p.39 / Chapter 3.2.4. --- Purification of polysaccharide-protein complex --- p.40 / Chapter 3.2.5. --- Molecular mass determination --- p.43 / Chapter 3.2.6. --- High performance liquid chromatography --- p.43 / Chapter 3.2.7. --- SDS-polyacrylamide gel electrophoresis --- p.44 / Chapter 3.2.8. --- Ultraviolet scanning --- p.44 / Chapter 3.2.9. --- Chemical analysis --- p.45 / Chapter 3.2.10. --- Experimental animals --- p.47 / Chapter 3.2.11. --- In vivo antitumor assay --- p.48 / Chapter 3.2.12. --- Safety tests --- p.49 / Chapter 3.2.13. --- Statistical analysis --- p.51 / Chapter 3.3. --- Results --- p.51 / Chapter 3.3.1. --- Extraction and purification --- p.51 / Chapter 3.3.2. --- Biochemical analysis --- p.52 / Chapter 3.3.3. --- Chemical analysis --- p.60 / Chapter 3.3.4. --- In vivo antitumor activity --- p.68 / Chapter 3.3.5. --- Safety evaluation --- p.68 / Chapter 3.4. --- Discussion --- p.75 / Chapter 3.5. --- Summary --- p.84 / Chapter Chapter 4. --- "Immunomodulating, Antitumor and other Biological Activities of Polysaccharide-Protein Complex (PSPC) from Tricholoma lobayense" --- p.85 / Chapter 4.1. --- Introduction --- p.85 / Chapter 4.2. --- Materials and methods --- p.87 / Chapter 4.2.1. --- Experimental animals --- p.87 / Chapter 4.2.2. --- Cultivation of tumor cells --- p.87 / Chapter 4.2.3. --- Preparation of peritoneal exudate cells and splenocytes --- p.87 / Chapter 4.2.4. --- Mitogenic response of T cells --- p.89 / Chapter 4.2.5. --- Responses of peritoneal exudate cells --- p.89 / Chapter 4.2.6. --- In vitro antitumor assay --- p.92 / Chapter 4.2.7. --- Transmission electron microscope --- p.93 / Chapter 4.2.8. --- Evaluation of other biological activities --- p.94 / Chapter 4.2.9. --- Statistical analysis --- p.99 / Chapter 4.3. --- Results --- p.99 / Chapter 4.3.1. --- Immunomodulating activity --- p.99 / Chapter 4.3.2. --- In vitro antitumor action --- p.107 / Chapter 4.3.3. --- Observation on tumor regression induced by PSPC --- p.107 / Chapter 4.3.4. --- Other biological actions --- p.112 / Chapter 4.4. --- Discussion --- p.121 / Chapter 4.4.1. --- Immunomodulating activity --- p.121 / Chapter 4.4.2. --- Antitumor activity --- p.125 / Chapter 4.4.3. --- Other biological activities --- p.127 / Chapter 4.5. --- Summary --- p.130 / Chapter Chapter 5. --- Induction of Gene Expression of Immunomodulating Cytokines by Polysaccharide-Protein Complex (PSPC) from Tricholoma lobayense --- p.132 / Chapter 5.1. --- Introduction --- p.132 / Chapter 5.2. --- Materials and methods --- p.135 / Chapter 5.2.1. --- Experimental animals --- p.135 / Chapter 5.2.2. --- Preparation of peritoneal exudate cells and splenocytes --- p.136 / Chapter 5.2.3. --- RNA extraction --- p.137 / Chapter 5.2.4. --- Reverse transcription- polymerase chain reaction --- p.137 / Chapter 5.2.5. --- Dot blot --- p.138 / Chapter 5.2.6. --- Hybridization --- p.141 / Chapter 5.3. --- Results --- p.142 / Chapter 5.3.1. --- mRNA phenotyping of cytokines and cytokine receptors in normal mice --- p.142 / Chapter 5.3.2. --- mRNA phenotyping of cytokines and cytokine receptors in tumor-bearing mice --- p.142 / Chapter 5.4. --- Discussion --- p.150 / Chapter 5.5. --- Summary --- p.153 / Chapter Chapter 6. --- General Discussion and Conclusion --- p.155 / REFERENCES --- p.159

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

Poon Shuk-ching. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 113-125). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / Abstract (Chinese Version) --- p.iv / Table of Contents --- p.vi / List of Tables --- p.x / List of Figures --- p.xi / List of Abbreviations --- p.xiii / Chapter Chapterl 1 --- ntroduction --- p.1 / Chapter Chapter 2 --- Literature Review --- p.5 / Chapter 2.1 --- Mushroom Polysaccharides From Basidiomycetes --- p.5 / Chapter 2.1.1 --- Antitumor and Immunomodulatory Activity --- p.6 / Chapter 2.1.2 --- Antiviral Activity --- p.9 / Chapter 2.1.3 --- Hypoglycermic Activity --- p.11 / Chapter 2.1.4 --- Free Radical Scavenging Activity --- p.11 / Chapter 2.2 --- Mushroom Dictyophora indusiata --- p.13 / Chapter 2.2.1 --- Nutritional Value --- p.13 / Chapter 2.2.2 --- Structural Characteristic of Dictyophora indusiata Polysaccharides --- p.14 / Chapter 2.2.3 --- Biological Activity --- p.16 / Chapter 2.3 --- In vivo Antitumor Study --- p.19 / Chapter 2.4 --- Induction of Cytokines Production in Immune System --- p.21 / Chapter 2.5 --- In vitro Antitumor Study --- p.25 / Chapter 2.6 --- Cell Cycle Regulation --- p.28 / Chapter Chapter 3 --- Materials & Methods --- p.34 / Chapter 3.1 --- Extraction --- p.34 / Chapter 3.1.1 --- Extraction of Dictyophora indusiata Polysaccharides --- p.34 / Chapter 3.1.2 --- Purification of Dictyophora indusiata Polysaccharides --- p.35 / Chapter 3.1.2.1 --- Preparation of DEAE-cellulose Ion Exchanger --- p.35 / Chapter 3.1.2.2 --- Fractionation --- p.35 / Chapter 3.2. --- Characterization of Dictyophora indusiata Polysaccharides --- p.39 / Chapter 3.2.1 --- Polysaccharide Content Determination --- p.39 / Chapter 3.2.2 --- Protein Content Determination --- p.39 / Chapter 3.2.3 --- Gas Chromatography (GC) --- p.40 / Chapter 3.2.4 --- Uronic Acid Content Determination --- p.42 / Chapter 3.2.5 --- High Performance Liquid Chromatography (HPLC) --- p.43 / Chapter 3.3 --- In vivo Studies --- p.44 / Chapter 3.3.1 --- Animals --- p.44 / Chapter 3.3.2 --- Maintenance of Sarcoma 180 Cell Line --- p.44 / Chapter 3.3.3 --- Effect of DI3 Fraction on Sarcoma 180 Solid Tumor --- p.45 / Chapter 3.3.4 --- Effect of DI3c Fraction on Tumor Necrosis Factor-Alpha (TNF-α) and Interleukin 2 (IL-2) Production --- p.47 / Chapter 3.3.4.1 --- Treatment of Mice --- p.47 / Chapter 3.3.4.2 --- Preparation of Mouse Serum --- p.47 / Chapter 3.3.4.3 --- Enzyme-linked Immunosorbent Assay (ELISA) for TNF-α Production --- p.48 / Chapter 3.3.4.4 --- Enzyme-linked Immunosorbent Assay (ELISA) for IL-2 Production --- p.49 / Chapter 3.4 --- In vitro Studies --- p.51 / Chapter 3.4.1 --- Maintenance of Cell Lines --- p.51 / Chapter 3.4.2 --- Effect on Cancer Cell Lines --- p.52 / Chapter 3.4.3 --- Cytotoxicity on Normal Cell Line --- p.52 / Chapter 3.4.4 --- Trypan Blue Exclusion Method --- p.53 / Chapter 3.4.5 --- MTT Assay --- p.54 / Chapter 3.4.6 --- BrdU Incorporation --- p.55 / Chapter 3.5 --- Statistical Analysis --- p.56 / Chapter Chapter 4 --- Results --- p.57 / Chapter 4.1 --- Extraction and Fractionation of Polysaccharides from Dictyophora indusiata --- p.57 / Chapter 4.1.1 --- Percentage Yield of Crude DI Polysaccharides --- p.57 / Chapter 4.1.2 --- Percentage Yield of DI3 Fractions --- p.57 / Chapter 4.2 --- Characterization of DI3 Fractions --- p.62 / Chapter 4.2.1 --- Polysaccharide and Protein Contents of DI3 Fractions --- p.62 / Chapter 4.2.2 --- Relative Monosaccharide and Uronic Acid Content in Different DI3 Fractions --- p.62 / Chapter 4.2.3 --- Estimated Molecular Weight of DI3 Fractions --- p.65 / Chapter 4.3 --- Antitumor Effect of Dictyophora indusiata Polysaccharides In vivo --- p.70 / Chapter 4.3.1 --- In vivo Antitumor Effect of Crude DI Polysaccharides --- p.70 / Chapter 4.3.2 --- In vivo Antitumor Effect of Various Fractions of DI3 --- p.70 / Chapter 4.3.3 --- Effect of DI3c on TNP-α and IL-2 Production in Mice --- p.78 / Chapter 4.4 --- In vitro Effects of DI3 Fractions on Cell Density and Viability on Normal and Cancer Cell Lines --- p.86 / Chapter 4.4.1 --- Effects of DI3 Fractions on Cell Density and Viability of Human Leukemic HL-60 and K-562 and Mouse Sarcoma 180 Cells --- p.86 / Chapter 4.4.2 --- Effects of DI3 Fractions on the Growth of Human Liver Cancer HepG2 and Normal Monkey Kidney Vero Cells --- p.86 / Chapter 4.4.3 --- Effect of DI3b Fraction on Proliferation of HL-60 Cells Determined by BrdU Incorporation --- p.94 / Chapter Chapter 5 --- Discussions --- p.96 / Chapter 5.1 --- Extraction and Characterization of DI3 Fractions --- p.96 / Chapter 5.2 --- Antitumor Effects of Dictyophora indusiata Polysaccharides --- p.101 / Chapter 5.3 --- Further Studies --- p.109 / Chapter Chapter 6 --- Conclusion --- p.111 / References --- p.113