Medicinal mushrooms are therapeutic agents in traditional folk medicines. Previous studies have shown that a number of biologically active compounds in medicinal mushrooms contributed therapeutic functions against many diseases. These compounds include mainly large molecular weight (MW) compounds such as polysaccharides, dietary fibre and lipids. Mushroom polysaccharides have attracted great attention in food and pharmacology fields because of their biological activities. Polysaccharides vary in molecular weight, degree of branching and conformational structure. It has been reported that fine structure, molecular weight, and conformation of polysaccharide influence biological activities. The incidence and prevalence of inflammatory bowel disease (IBD) have been increasing worldwide, which is characterized by chronic inflammation of the gastrointestinal tract but without satisfactory treatment. Although there are many studies for the immuno-pharmacological activity of mushroom polysaccharides, their intestinal anti-inflammatory property has not been investigated sufficiently. Therefore, it is very important to elucidate whether there is the relationship among the MW, structure and anti-inflammatory activity of polysaccharide in IBD. Firstly, an exopolysaccharide from a mycelial culture of S. commune was obtained by isolation and purification using DEAE-52 cellulose and Sephadex G-150 column chromatography. The structure, conformation and chemical properties were investigated, including elemental compositions, MW, monosaccharide compositions, fourier transform infrared spectrum, thermogram analysis, nuclear magnetic resonance (NMR) spectrum, circular dichroism (CD) study, methylation analysis, and scanning electron microscope (SEM). The findings indicate that the exopolysaccharide is a homogeneous protein-bound heteropolysaccharide carrying molecular weight of 2900 kDa with a β-type glycosidic linkage. It belongs to a kind of β-(13)-D-glucans consisting of a backbone of β-(13)-linked glucose residues branched with (14) and (16)-β-D-glucopyranosyl residues on main-chain residues. The elemental analysis of this exopolysaccharide discover the element compositions as: C, 25.84%; H, 5.45%; and N, 0.65%. The total carbohydrate, protein and uronic acid contents of exopolysaccharide is 89.0%, 2.20% and 7.52%, respectively. In addition, lipopolysaccharide (LPS) was not detected in the exopolysaccharide. Glucose is the main monosaccharide structural unit in this exopolysaccharide, the content is 57.5%. The degradation temperature of exopolysaccharide is 278.9°C from the thermogram analysis curve. This exopolysaccharide looks like thin film with smooth and glittering surface in SEM photography. It is clear from these images that the exopolysaccharide is linear in structure and branched and coiled in aqueous solution. With these extraction, the preliminary anti-inflammatory activity of S. commune exopolysaccharide was conducted by inhibiting the production of nitric oxide (NO), activity of inducible nitric oxide synthase (iNOS) and activity of 5-lipoxygenase (5-LOX) from RAW 264.7 macrophages. The results showed that exopolysaccharide significantly inhibit LPS-induced iNOS expression levels in a dose-dependent manner(p < 0.05). It inhibits the production of 5-LOX in cells, but not in dose-dependence. Further, in dextran sulfate sodium (DSS)-induced colitis model, the results showed that exopolysaccharide attenuated body weight loss, diarrhea, fecal blood, and the shortening of colon and improved histological changes. Furthermore, exopolysaccharide treatment would reduce NO production and some cytokines' secretion such as IL-4 and IL-17A. These results indicate that exopolysaccharide might be exploited as an effective anti-inflammatory agent for application in IBD. Secondly, ultrasound technology was applied to modify the physicochemical properties (MW and viscosity) of this fungal exopolysaccharide, and fractions of different MWs were obtained through ultrasonic degradation method. Effect of the MW degradation, viscosity and anti-inflammatory property of exopolysaccharide under ultrasonic treatment were optimized with response surface methodology. The best ultrasonic treatment parameters were obtained with a three-variable-three-level Box-Behnken design. The optimized conditions for efficient anti-inflammatory activity include: Initial concentration - 0.4%; ultrasonic power - 600 W; and duration of ultrasonic treatment - 9 min. Under these conditions, the NO inhibition rate is 95 ± 0.03% which agreed closely with the predicted value (96%). Average MW of exopolysaccharide decreased after ultrasonic treatments, but no significant change in the preliminary structure by infrared spectroscopy analysis. The viscosity of degraded exopolysaccharide dropped compared with native exopolysaccharide. The results suggest that ultrasound technology is an effective approach to reduce the MW of exopolysaccharide. Our results also showed that exopolysaccharide from S. commune was degraded into three fractions (low, medium, and high MW) by ultrasonic treatment. The changes of MW, atomic force microscope morphology, X-ray diffraction, particle size distribution and viscosity analysis indicate the triple helical structure of exopolysaccharide was dissociated into single helical structure and random coiled structure by breaking of inter- and intramolecular hydrogen bonds. The medium and high MW exopolysaccharide had the mixture of triple helix and single helix conformation. Moreover, the low MW exopolysaccharide exhibit random coiled conformation. As for their anti-inflammatory effect in DSS-induced colitis mice model, the results showed that medium and high MW exopolysaccharide significantly recovered DSS-induced colitis in body weight loss, shortening of colon lengths, colon weight loss, diarrhea and rectal bleeding, histological score, myeloperoxidase (MPO) activity, NO and cytokines (IFN-γ, IL-10 and IL-17) production in inflamed tissues. Moreover, exopolysaccharide with medium and high MW reduced DSS-induced infiltration of macrophages. These results showed that medium and high MW exopolysaccharide had intestinal anti-inflammatory activity. The degraded exopolysaccharide with medium and high MW had a triple and single-helical structure. These results suggested that the intestinal anti-inflammatory activity of exopolysaccharide from S. commune is related to both helical structure and MW.

Abstract not available

Herbs have been the basis for medical treatments through much of human history, and even now such herbalism is still widely practiced around the world. Most frequently and traditionally, water is used as the extraction solvent for preparing medicinal herbs to generate decoction or infusion for medicinal purpose. In other words, in most cases, multiple chemical components in water extracts should be responsible for therapeutic (toxic and side, if any) effects of medicinal herbs. Phytochemical analysis of water extracts for quality control of medicinal herbs is therefore important to ensure their safeties and efficacies. Unfortunately, however, it is not given enough attention in the modern research whereas the relative current studies are intensively focused on organic solvent-extracts of medicinal herbs. In this project, analysis of medicinal herbs’ water extracts is thus focused. Various analytical approaches have been exhaustively developed for qualitative and quantitative analysis of chemicals in water extracts of medicinal herbs. However, many research challenges in methodology still exist. Polysaccharides and small molecules are two most important kinds of chemcials in water extracts of medicinal herbs, so they also widely regarded as markers for quality evaluation. For analysis of small molecules, the levels of quantitative determination are always far unsatisfactory, normally less than 10%. For analysis of polysaccharides, the existed problems are even more serious in both sample preparation and chemical analysis. Ethanol precipitation is always the first step for crude polysaccharide preparation. But it is just directly used without optimization and its capacity has never been evaluated. Following that, chemical analysis of natural polysaccharide also suffers severe methodological bottlenecks and many drawbacks occurre in qualitative and quantitative characterization. Besides, polysaccharides and small molecules in medicinal herbs are always individually investigated but hardly studied together before. Concerning these issues, here several approaches and stratigies were accordingly proposed to improve the current situations using decoctions of some traditional Chinese medicines (TCMs) as the research objects and examples. In detail, first, a quantitative method was developed for quality evaluation of Huang-Lian-Jie-Du-Tang. In this study, quantitative levels of small molecules were greatly improved, compared with the current analogous studies for quality evaluation of medicinal herbs. Then, shifting to polysaccharides, availability of ethanol precipitation for natural polysaccharide precipitation was critically evaluated. Parameters which could affect the ethanol precipitation results, such as structural features, molecular size of polysaccharide, and ethanol concentration were systematically investigated. Successively, a novel and rapid HPGPC-based strategy for quality control of saccharide-dominant medicinal herbs was proposed using Dendrobium officinale as the example. Polysaccharides in the decoction of Dendrobium officinale were qualitatively and quantitatively determined. The methodological superiority of the developed method compared with conventional approaches was highlighted. To facilitate this study, research on chemistry, bioactivity and quality control of Dendrobium was systematically reviewed in advance. After that, small molecules and polysaccharides in in Angelicae Sinensis Radix and Chuanxiong Rhizoma were compared together. Lastly, effects of ginseng polysaccharides on the in vivo pharmacokinetics of ginsenoside Rg1 on induced immunosuppressive model rats was investigated to provide a chemically holistic view for Du-Shen-Tang. By these studies, the above mentioned predicament in chemical analysis on both small molecuels and polysaccharides in water extracts of medicinal herbs were methodologically improved to varying degrees. Concerning small molecules and polysaccharides from multiple perspectives, the successive studies are helpful for enhancing quality evaluation and scientific understanding of medicinal herbs’ decoctions.

Lai Kin Ming Connie. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 121-141). / Abstracts in English and Chinese. / Chapter Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Introduction on growth cycle of mushroom --- p.1 / Chapter 1.2 --- Literature review of mushroom biological activities --- p.3 / Chapter 1.2.1 --- Various bioactivities of mushroom --- p.3 / Chapter 1.2.2 --- Components responsible for various bioactivities of mushrooms --- p.3 / Chapter 1.3 --- Mushroom polysaccharides and polysaccharide-protein complexes --- p.5 / Chapter 1.3.1 --- Polysaccharides important for antitumor effects --- p.5 / Chapter 1.3.2 --- Polysaccharide-protein complexes important for antitumor effects --- p.7 / Chapter 1.4 --- Structure-function relationship of antitumor activities of polysaccharides --- p.8 / Chapter 1.4.1 --- Effect of molecular mass --- p.8 / Chapter 1.4.2 --- Effect of linkages --- p.9 / Chapter 1.4.3 --- Effect of degree of branching --- p.9 / Chapter 1.4.4 --- Effect of conformation --- p.10 / Chapter 1.5 --- Immunomodulatory effects of mushroom polysaccharides and polysaccharide-protein complexes --- p.11 / Chapter 1.5.1 --- Immunomodulatory effects of polysaccharides --- p.11 / Chapter 1.5.1.1 --- Bioactive polysaccharides in Lentinus edodes --- p.11 / Chapter 1.5.1.2 --- Bioactive polysaccharides in Ganoderma lucidum --- p.12 / Chapter 1.5.2 --- Immunomodulatory effects of polysaccharide-protein complexes --- p.12 / Chapter 1.5.2.1 --- Bioactive polysaccharide-protein complexes in Trametes versicolor --- p.13 / Chapter 1.5.3 --- Immunotherapeutic effects of mushroom polysaccharides --- p.14 / Chapter 1.6 --- Cell cycle and apoptosis --- p.14 / Chapter 1.6.1 --- Introduction of cell cycle --- p.14 / Chapter 1.6.2 --- Cell cycle regulation --- p.15 / Chapter 1.6.3 --- Antitumor effects through apoptotic gene regulation --- p.17 / Chapter 1.7 --- Mushroom sclerotium with antitumor activity --- p.20 / Chapter 1.7.1 --- Literature review on Pleurotus tuber-regium --- p.20 / Chapter 1.7.2 --- Literature review on Poria cocos --- p.22 / Chapter 1.7.3 --- Literature review on Polyporus rhinocerus --- p.23 / Chapter 1.8 --- Objectives --- p.23 / Chapter Chapter 2. --- Materials and Methods --- p.25 / Chapter 2.1 --- Materials --- p.25 / Chapter 2.1.1 --- Mushroom sclerotia --- p.25 / Chapter 2.1.2 --- Animal Model --- p.25 / Chapter 2.1.3 --- Cell lines --- p.27 / Chapter 2.2 --- Methods --- p.28 / Chapter 2.2.1 --- Extraction Scheme for mushroom sclerotia --- p.28 / Chapter 2.2.1.1 --- Hot water extraction only --- p.28 / Chapter 2.2.1.2 --- Sequential extraction scheme --- p.28 / Chapter 2.2.2 --- Measurement of monosaccharide profile --- p.31 / Chapter 2.2.2.1 --- Acid Depolymerisation --- p.31 / Chapter 2.2.2.2 --- Neutral Sugar Derivatization --- p.31 / Chapter 2.2.2.3 --- Gas Chromatography (GC) --- p.32 / Chapter 2.2.3 --- High Pressure Liquid Chromatography (HPLC) --- p.33 / Chapter 2.2.3.1 --- Size exclusion chromatography --- p.33 / Chapter 2.2.3.2 --- Anion exchange chromatography --- p.34 / Chapter 2.2.4 --- Linkage analysis by methylation --- p.34 / Chapter 2.2.4.1 --- Preparation of partially methylated polysaccharides --- p.34 / Chapter 2.2.4.2 --- Preparation of partially methylated alditol acetates (PMAAs) --- p.37 / Chapter 2.2.4.3 --- Gas chromatography-Mass spectrometry (GC-MS) analysis --- p.37 / Chapter 2.2.5 --- Determination of total sugar by phenol-sulphuric acid Method --- p.38 / Chapter 2.2.6 --- Determination of acidic sugars by measurement of uronic acid content --- p.39 / Chapter 2.2.7 --- Determination of protein content by Lowry-Folin method --- p.40 / Chapter 2.2.8 --- Chemical modification by carboxymethylation --- p.41 / Chapter 2.2.9 --- In vitro antitumor assay --- p.41 / Chapter 2.2.9.1 --- Trypan blue exclusion assay --- p.42 / Chapter 2.2.9.2 --- MTT Assay --- p.42 / Chapter 2.2.10 --- Cell cycle analysis by Flow Cytometry --- p.43 / Chapter 2.2.11 --- In vivo antitumor and immunomodulatory assay --- p.44 / Chapter 2.2.11.1 --- Measurement on tumor growth --- p.44 / Chapter 2.2.11.2 --- Blood sampling for immunostimulatory effects --- p.45 / Chapter 2.2.12 --- Mouse Cytokine Array --- p.45 / Chapter 2.2.13 --- Quantification of Mouse IL-13 by ELISA --- p.46 / Chapter 2.2.14 --- Enumeration of peritoneal cells --- p.47 / Chapter 2.2.15 --- Enumeration of splenocytes --- p.49 / Chapter 2.2.16 --- Statistical methods --- p.50 / Chapter Chapter 3. --- Results and Discussion --- p.51 / Chapter 3.1 --- Yield of crude mushroom sclerotial extracts --- p.51 / Chapter 3.2 --- Chemical composition of crude mushroom sclerotial extracts --- p.57 / Chapter 3.2.1 --- Total carbohydrate content --- p.57 / Chapter 3.2.2 --- Uronic acid content --- p.58 / Chapter 3.2.3 --- Soluble protein content --- p.58 / Chapter 3.3 --- Monosaccharide profiles of mushroom sclerotial extracts by GC --- p.60 / Chapter 3.4 --- Chromatographic analyses of mushroom sclerotial extracts --- p.65 / Chapter 3.4.1 --- Molecular weight profile by size exclusion chromatography (SEC) --- p.65 / Chapter 3.4.2 --- Charge distribution by ion exchange chromatography (IEC) --- p.73 / Chapter 3.5 --- Antitumor effects of mushroom sclerotial extracts from hot water extraction alone --- p.73 / Chapter 3.5.1 --- In vitro antiproliferative study by HL-60 --- p.73 / Chapter 3.5.2 --- In vitro antiproliferative study by MCF-7 --- p.74 / Chapter 3.5.3 --- In vivo antitumor study by BALB/c mice --- p.75 / Chapter 3.6 --- Antitumor effects of extracts from sequential extraction scheme --- p.76 / Chapter 3.6.1 --- In vitro antiproliferative study by HL-60 --- p.76 / Chapter 3.6.2 --- In vitro antiproliferative study by MCF-7 --- p.78 / Chapter 3.6.3 --- In vivo antitumor study by BALB/c mice --- p.80 / Chapter 3.7 --- Comparison of in vitro and in vivo activities of mushroom sclerotial extracts --- p.82 / Chapter 3.8 --- Dose-response relationship of hot water extract from PR on cancer cell lines --- p.85 / Chapter 3.8.1 --- In vitro dose-response antiproliferation of PR-W and PR-HWE on HL-60 --- p.85 / Chapter 3.8.2 --- In vitro dose-response antiproliferation of PR-W on K562 and S180 --- p.88 / Chapter 3.8.3 --- In vivo dose-response relationship of PR-W on S180 --- p.91 / Chapter 3.9 --- Flow cytometric analysis of PR-W on cancer cell lines --- p.92 / Chapter 3.9.1 --- Antiproliferative effect of PR-W on HL-60 --- p.92 / Chapter 3.9.2 --- Antiproliferative effect of PR-W on K562 --- p.95 / Chapter 3.9.3 --- Proposed mechanisms of cell cycle arrest by PR-W --- p.98 / Chapter 3.10 --- Host-mediated antitumor mechanism of PR-W --- p.100 / Chapter 3.10.1 --- Mouse cytokine array --- p.100 / Chapter 3.10.2 --- Quantification of IL-13 by ELISA --- p.105 / Chapter 3.10.3 --- Immunostimulatory effects of PR-W on mice --- p.109 / Chapter 3.11 --- Correlation between antitumor activity and structure of mushroom sclerotial extract from hot water extraction alone --- p.114 / Chapter Chapter 4. --- Conclusions and Future works --- p.118 / List of References --- p.121 / Related Publications --- p.142

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&lt;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&lt;0.05) on human normal spleen monocyte/macrophage cell, MD. Moreover, PRSon was shown to have a significant increase (p&lt;0.05) in the growth of human natural killer cells, NK-92M1; however, PTRW showed a significant inhibition (p&lt;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&lt;0.05) of Dectin-1 expression on NK-92MI cells incubated with PTRW. On the other hand, PTRW caused a significant decrease ( p&lt;0.05) of Dectin-1 expression while PRSon showed a significant increase (p&lt;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&lt;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&lt;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.

Radix Astragali (the dried root of Astragalus membranaceous (Fisch) Bge.) and Dendrobii Officinalis Caulis (the dried stem of Dendrobium officinale Kimura et Migo) are two traditional Chinese tonic herbs. They are commonly used in the formula with other Chinese herbs for tonifying Qi, nourishing Yin, and treating various kinds of diseases, such as cancer, diabetes, inflammation, etc. The polysaccharides are considered the majority of the chemical components of decoction boiled from a formula including these two medicinal herbs. The previous study showed that the polysaccharides isolated from Radix Astragali (named RAP) and Dendrobii Officinalis Caulis (named DOP) have various pharmacological activities and most of their activities are closely related to their immunomodulating effects. Nonetheless, the exact mechanism of their immunomodulating effects, especially on macrophages is not known clearly. In the current study, we have conducted a comprehensive investigation of the bioactive properties and molecular mechanism of immunomodulating activities of DOP and RAP. We aimed to clarify the molecular immunomodulating mechanism of RAP on macrophages and the actual anti-fatigue activity of DOP in vivo. Results can be summarized as follows: RAP itself did not have any cytotoxic effect on mouse mammary carcinoma 4T1 cells, but it significantly enhanced cytotoxicity of the supernatant of RAW264.7cells on 4T1 cells. Furthermore, RAP enhanced the production of NO and cytokines in RAW264.7 cells, and significantly up-regulated gene expressions of TNF-α, IL-6, iNOS. All these bioactivities were blocked by the inhibitor of TLR4 (Toll-like receptor 4), suggesting that TLR4 is a receptor of RAP and mediates its immunomodulating activity. Further analyses demonstrated that RAP rapidly activated TLR4-related MAPKs, including phosphorylated ERK, phosphorylated JNK, and phosphorylated p38, and induced translocation of NF-κB as well as degradation of IκB-α. In addition, RAP induced higher gene expression of M1 marker, including iNOS, IL-6, TNF-a, CXCL10, compared with those of control group. RAP-induced BMDMs were polarized from M2 to M1 phenotypes. RAP stimulated RAW264.7 cells to express Notch1, Notch2, Jaddge1, Dll1 and SOCS3. Notch signaling pathway played an important role in the RAP-induced polarization of M1 phenotype macrophages. The RAP-induced BMDMs exhibited anti-cancer effect when they were transplanted with 4T1 cells together in vivo and it decreased tumor volume and tumor weight. DOP, the authentication marker of Dendrobii Officinalis Caulis, has immunomodulating activity in macrophage cell line RAW 264.7. DOP enhanced cell proliferation, TNF-α secretion, and phagocytosis in a dose-dependent manner. It induced the proliferation of lymphocytes alone and with mitogens. For further study the anti-fatigue effect of DOP in vivo, the weight-loaded swimming test was used, because it is an effective method for evaluation of the extent of fatigue. The results indicated that DOP treatment significantly increased the swimming endurance time, body weight, and food intake, compared to the positive control Rhodiola rosea extract. Moreover, the weight-loaded swimming test decreased the levels of glycogen in gastrocnemius muscle, SOD, GSH-Px in serum, and increased the levels of LDH, BUN, MDA, CK, TG, and LD in serum. All of these indicators of fatigue were inhibited to a certain extent by both DOP and Rhodiola rosea extract, and DOP's effects are stronger. Furthermore, DOP-feeding mice showed significantly increased cell variability of T lymphocytes and B lymphocytes, compared with control mice. In conclusion, RAP may induce cytokine production of RAW264.7 cells through TLR4-mediated activation of MAPKs and NF-κB. RAP-induced BMDMs were polarized from M2 to M1 phenotypes through Notch signaling pathway. The unique and dominant polysaccharide DOP is proven to be major, active polysaccharide markers of D. officinale, and showed stronger anti-fatigue activity than Rhodiola rosea extract. As such, DOP has promising potential for pharmaceutical development into anti-fatigue health product.

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

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

by Liu Chi. / "July, 2003." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (p. 270-283). / 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 Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.