Li Qian. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 102-117). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.vi / List of Abbreviations --- p.ix / List of tables and figures --- p.ix / Content --- p.x / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1 --- Traditional Chinese Medicine: Agrimony --- p.1 / Chapter 1.2 --- Hepatocellular carcinoma (HCC) and its risk factors --- p.4 / Chapter 1.3 --- Basic concepts relevant to cancer prevention --- p.6 / Chapter 1.3.1 --- Multistage process of carcinogenesis --- p.6 / Chapter 1.3.2 --- Chemical carcinogenesis --- p.7 / Chapter 1.3.3 --- Possible chemopreventive strategies --- p.8 / Chapter 1.3.4 --- Phase I and phase II systems in chemical carcinogenesis --- p.10 / Chapter Chapter 2 --- Materials and methods --- p.12 / Chapter 2.1 --- Preparation of aqueous extract of Agrimonia pilosa --- p.12 / Chapter 2.2 --- In vivo study --- p.13 / Chapter 2.2.1 --- Animal model for hepatocarcinogenesis --- p.13 / Chapter 2.2.1.1 --- Chemical carcinogens --- p.13 / Chapter 2.2.1.2 --- Animals --- p.16 / Chapter 2.2.1.3 --- Animal treatment and sacrifice --- p.17 / Chapter 2.2.2 --- Histological and immunohistochemical study --- p.20 / Chapter 2.2.3 --- Preparation of liver homogenates and microsomes from rat --- p.23 / Chapter 2.2.4 --- Determination of protein concentration --- p.24 / Chapter 2.2.5 --- COX-2 Activity Assay --- p.25 / Chapter 2.2.6 --- Cytochrome P450 2E1 Assay --- p.26 / Chapter 2.2.7 --- Spectrophotometry Assay for GST --- p.28 / Chapter 2.2.8 --- Isolation of total RNA from liver homogenate --- p.29 / Chapter 2.2.9 --- Semi-quantitative RT-PCR analysis --- p.32 / Chapter 2.3 --- In vitro study --- p.36 / Chapter 2.3.1 --- Cell cultures --- p.36 / Chapter 2.3.2 --- Cytotoxicity assay - Neutral Red Assay --- p.38 / Chapter 2.3.3 --- Cell cycle distribution analysis by flow cytometry --- p.39 / Chapter 2.3.4 --- DNA fragmentation --- p.40 / Chapter Chapter 3 --- Results --- p.43 / Chapter 3.1 --- In vivo study --- p.43 / Chapter 3.1.1 --- Body weight and relative liver weight --- p.43 / Chapter 3.1.2 --- Gross Morphological changes --- p.46 / Chapter 3.1.3 --- Hematoxylin & Eosin (H&E) staining for histological detection --- p.50 / Chapter 3.1.4 --- Effect of AP on DEN-CCl4-induced GST-P positive foci formation and GST-P mRNA expression --- p.60 / Chapter 3.1.5 --- Effects of AP on COX-2 --- p.72 / Chapter 3.1.6 --- Effects of AP on phase I and phase II enzymes --- p.76 / Chapter 3.2 --- In vitro study --- p.80 / Chapter 3.2.1 --- Effects of AP on proliferation of H4IIE cells detected by Neutral Red Assay --- p.80 / Chapter 3.2.2 --- Assessment of cell cycle distribution by flow cytometry --- p.82 / Chapter 3.2.3 --- DNA Fragmentation Assay --- p.88 / Chapter Chapter 4 --- Discussion --- p.90 / Chapter 4.1 --- In vivo study --- p.90 / Chapter 4.1.1 --- Morphological changes during the induction of hepatocarcinogenesis --- p.90 / Chapter 4.1.2 --- Effects of AP on GST-P foci and its mRNA --- p.91 / Chapter 4.1.3 --- Effects of AP on COX-2 enzyme activity and mRNA expression --- p.93 / Chapter 4.1.4 --- Modulation effects of AP on CYP2E1 and GST enzyme activity --- p.95 / Chapter 4.2 --- In vitro study: effects of AP on cancer cell proliferation --- p.97 / Chapter 4.3 --- Summary --- p.99 / References --- p.102

Zhang Yongwen. / "February 2003." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (p. 174-188). / 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.

Conclusions. SBL showed its efficacy in treating the animal model of allergic rhinitis. Its mechanisms may be related to its suppressive action on PCA reaction, the production of TXB2 and the expression of eNOS, as well as its modulation of cytokines, including IL-4, IL-6, IL-8, GM-CSF and TNF-alpha, release from mast cells. The clinical trial showed that SBL had more beneficial action on the quality of life, in comparison to the placebo, in the domains of RE and BP. Some symptoms evaluations of PAR patients, including GF, NB and SF were more markedly improved in the SBL group when compared with the placebo group. Furthermore, the use of SBL, with the study dose and treatment period, was safe. However, the accurate efficacy and mechanisms of SBL are largely unknown and need further investigation. (Abstract shortened by UMI.) / Introduction. Although great progress in treatment of allergic rhinitis have made in recent years, remarkably increasing prevalence and cost in epidemiology studies strongly suggest the difficulties in the management of allergic rhinitis. Shi-Bi-Lin (SBL) is a formula modified from the traditional Chinese herbal formula Cang-Er-Zi-San (CEZS) and a classic European formula SinupretRTM. CEZS has been used for the treatment of allergic rhinitis for several centuries in East Asia communities, and SinupretRTM has been used in treating paranasal sinusitis and rhinitis widely in Europe for decades. However, its therapeutic mechanisms remain unclear. We examined the efficacy and the possible mechanism of SBL in an animal model of allergic rhinitis and in cell culture study using Human Mast Cell Line (HMC-1) and Peripheral Blood Mononuclear Cells (PBMC). In addition, a clinical trial was conducted to examine its clinical efficacy and safety. / Results. In the animal study, SBL showed a potent effect in relieving the symptoms of nasal obstruction, sneezing and nasal scratching (P<0.05 or P<0.01), but had no convincing effect in decreasing the nasal discharge (P>0.05). In PCA test, IgG1 increased in a modest manner in the SBL-treated group when compared with the sham group (P<0.05 or P<0.01). Eosinophil infiltration and the expression of eNOS in nasal mucosa, but not iNOS, were obviously lower in the SBL treated group (P<0.05 or P<0.01) in comparison to the sham group. The levels of thromboxane B (TXB)2 in the nasal lavage fluid, but not histamine and peptide leukotrienes (p-LTs), showed significantly lower than that of the sham group (P<0.05). In vitro study showed that SBL modulated the cytokines, including interleukin (IL)-4, IL-6, IL-8, Granulocyte/Macrophage Colony-Stimulating Factor (GM-CSF) and tumor necrosis factor (TNF)-alpha, release from human mast cell line (HMC-1). However, the mRNA expressions of these cytokines were not significantly altered. As the controls, dexamethasone, desloratadine and budesonide had more potently inhibitory effects on cytokines release from HMC-1. The component herbs generally had stimulatory effects on the cytokine release from HMC-1 and variable effects on PBMC. In the clinical trial, a total of 84 patients were recruited in the clinical trial and 77 of them completed the trial. Although no significant differences of each domain between the SBL and placebo groups were detected, findings supported the efficacy of SBL were obtained. / by Zhao Yu. / "July 2005." / Advisers: C. A. Van Hasselt; Ping-Chung Leung; Kong-Sang Woo. / Source: Dissertation Abstracts International, Volume: 67-01, Section: B, page: 0172. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references. / 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. / Abstract in English and Chinese. / School code: 1307.

by Zhang Yanbo. / "December 2003." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (p. 163-185). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Song Jingzheng. / "June 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 170-186). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

by Ngan Fai Ngor Karenda. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (leaves 128-134). / Acknowledgements --- p.i / Abstract --- p.ii / Table of Contents --- p.iii / Abbreviations --- p.viii / Chapter Chapter 1 --- Authentication of Chinese Medicinal Herbs / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.2 --- Traditional Identification of Chinese Herbs / Chapter 1.2.1 --- Morphology --- p.3 / Chapter 1.2.2 --- Histology --- p.4 / Chapter 1.2.3 --- Chemical Analysis --- p.4 / Chapter 1.2.4 --- Proteins and Isozymes --- p.6 / Chapter 1.3 --- Molecular Technology in Authentication / Chapter 1.3.1 --- Restriction Fragment Length Polymorphism (RFLP) --- p.6 / Chapter 1.3.2 --- Polymerase Chain Reactions (PCRs) / Chapter 1.3.2.1 --- Random-Primed PCRs --- p.8 / Chapter 1.3.2.2 --- Simple Sequence Repeats --- p.10 / Chapter 1.3.2.3 --- Amplified Fragment Length Polymorphism (AFLP) --- p.11 / Chapter 1.4 --- Objectives and Strategies of the Study --- p.13 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Reagents and Buffers / Chapter 2.1.1 --- Buffers for Total DNA Extraction --- p.15 / Chapter 2.1.2 --- Reagents for Agarose Gel Electrophoresis --- p.16 / Chapter 2.1.3 --- Reagents for Polyacrylamide Gel Electrophoresis --- p.17 / Chapter 2.1.4 --- Reagents for Plasmid and Single-Stranded DNA Preparation --- p.17 / Chapter 2.1.5 --- Media for Bacterial Culture --- p.19 / Chapter 2.1.6 --- Reagents for Preparation of Competent Cells --- p.20 / Chapter 2.2 --- DNA Isolation / Chapter 2.2.1 --- Sample Preparation --- p.21 / Chapter 2.2.2 --- Cetyl triethylammonium bromide (CTAB) Extraction --- p.21 / Chapter 2.2.3 --- Cesium Chloride Gradient Ultracentrifugation --- p.21 / Chapter 2.3 --- Phenol/Chloroform Extraction --- p.22 / Chapter 2.4 --- Ethanol Precipitation --- p.23 / Chapter 2.5 --- DNA Concentration/Purity Estimation --- p.23 / Chapter 2.6 --- Random-Primed Polymerase Chain Reactions / Chapter 2.6.1 --- Random Amplified Polymorphic DNA (RAPD) --- p.24 / Chapter 2.6.2 --- Arbitarily-Primed Polymerase Chain Reaction (AP-PCR) --- p.24 / Chapter 2.7 --- rDNA Amplification --- p.24 / Chapter 2.8 --- Agarose Gel Electrophoresis of DNA --- p.25 / Chapter 2.9 --- Purification of rDNA / Chapter 2.9.1 --- from Agarose Gel using Geneclean II Kit (Bio 101 Inc.) --- p.25 / Chapter 2.9.2 --- using Microspin´ёØ Columns --- p.26 / Chapter 2.10 --- Preparation of Escherichia coli Competent Cells --- p.26 / Chapter 2.11 --- Ligation and Transformation of Escherichia coli --- p.27 / Chapter 2.12 --- Isolation of Plasmid DNA --- p.27 / Chapter 2.13 --- Screening of Plasmid DNA by Restriction Digestion --- p.28 / Chapter 2.14 --- Isolation of Plasmid DNA / Chapter 2.14.1 --- Minipreparation of Plasmid using Magic´ёØ Miniprep DNA Purification Kit from Promega --- p.28 / Chapter 2.14.2 --- Megapreparation of Plasmid using Qiagen-tip100 --- p.28 / Chapter 2.15 --- Single-Stranded DNA Preparation / Chapter 2.15.1 --- Transfection --- p.29 / Chapter 2.15.2 --- Single-Stranded DNA Isolation --- p.29 / Chapter 2.16 --- DNA Sequencing / Chapter 2.16.1 --- Plasmid Sequencing using T7 Sequencing Kit --- p.30 / Chapter 2.16.2 --- Cycle Sequencing from PCR Products --- p.30 / Chapter 2.16.3 --- Cycle Sequencing from PCR Products or Plasmid --- p.31 / Chapter 2.16.4 --- DNA Sequencing Electrophoresis --- p.31 / Chapter Chapter 3 --- Studies of Panax Species by Random-Primed PCRs / Chapter 3.1 --- Introduction --- p.34 / Chapter 3.2 --- Materials and Methods / Chapter 3.2.1 --- Plant Materials --- p.39 / Chapter 3.2.2 --- DNA Extraction and Random-Primed PCRs --- p.39 / Chapter 3.2.3 --- Data Analysis --- p.39 / Chapter 3.3 --- Results and Discussion / Chapter 3.3.1 --- DNA Isolation --- p.40 / Chapter 3.3.2 --- DNA Fingerprinting --- p.41 / Chapter 3.3.3 --- Relationship between the Six Panax Species --- p.45 / Chapter Chapter 4 --- Studies of Acorus by Random-Primed PCRs / Chapter 4.1 --- Introduction --- p.48 / Chapter 4.2 --- Materials and Methods / Chapter 4.2.1 --- Plant Materials --- p.49 / Chapter 4.2.2 --- DNA Extraction and Random-Primed PCRs --- p.50 / Chapter 4.3 --- Results and Discussion / Chapter 4.3.1 --- Acorus DNA --- p.50 / Chapter 4.3.2 --- Reproducibility of Random-Primed PCRs --- p.51 / Chapter 4.3.3 --- DNA Fingerprinting --- p.53 / Chapter Chapter 5 --- Studies of Epimedium by Random-Primed PCRs / Chapter 5.1 --- Introduction --- p.70 / Chapter 5.2 --- Materials and Methods / Chapter 5.2.1 --- Plant Materials --- p.71 / Chapter 5.2.2 --- DNA Extraction and Random-Primed PCRs --- p.71 / Chapter 5.3 --- Results and Discussion / Chapter 5.3.1 --- DNA Extraction --- p.71 / Chapter 5.3.2 --- DNA Fingerprinting --- p.72 / Chapter Chapter 6 --- Application of AP-PCR in Commercial Ginseng Products / Chapter 6.1 --- Introduction --- p.90 / Chapter 6.2 --- Materials and Methods / Chapter 6.2.1 --- Materials --- p.91 / Chapter 6.2.2 --- DNA Extraction and Random-Primed PCRs --- p.91 / Chapter 6.2.3. --- Data Analysis --- p.91 / Chapter 6.3 --- Results and Discussion / Chapter 6.3.1 --- DNA Isolation --- p.92 / Chapter 6.3.2 --- AP-PCR Analysis --- p.93 / Chapter Chapter 7 --- Ribosomal DNA as a Marker in Authentication of Panax Species / Chapter 7.1 --- Introduction --- p.99 / Chapter 7.2 --- Materials and Methods / Chapter 7.2.1 --- Plant Materials --- p.100 / Chapter 7.2.2 --- DNA Extraction and rDNA Amplification --- p.101 / Chapter 7.2.3 --- rDNA Sequencing --- p.101 / Chapter 7.2.4 --- Generation of Restriction Fragment Length Polymorphisms / Chapter 7.2.4.1 --- Restriction Digestion of rDNA Fragment --- p.102 / Chapter 7.2.4.2 --- Polyacrylamide Gel Electrophoresis (PAGE) --- p.103 / Chapter 7.2.4.3 --- Silver Staining for Nucleic Acids --- p.103 / Chapter 7.2.5 --- Data Analysis --- p.104 / Chapter 7.3 --- Results and Discussion / Chapter 7.3.1 --- rDNA Amplification and Plasmid Isolation --- p.104 / Chapter 7.3.2 --- rDNA Sequencing / Chapter 7.3.2.1 --- Sequence Comparison between the Six Panax species and the Two Adulterants --- p.107 / Chapter 7.3.3 --- Restriction Fragment Length Polymorphisms / Chapter 7.3.3.1 --- Restriction Profiles between Ginsengs and their Adulterants --- p.113 / Chapter 7.3.3.2 --- Restrciton Profiles of Ginsengs from Different Sources --- p.118 / Chapter 7.3.4 --- Panax Phylogeny --- p.121 / Chapter Chapter 8 --- General Discussion / Chapter 8.1 --- Advantages of Random-Primed PCRs --- p.124 / Chapter 8.2 --- Weaknesses of the Random-Primed PCRs --- p.125 / Chapter 8.3 --- Molecular Markers for Phylogenetic Studies --- p.126 / Chapter 8.4 --- Specific PCR-RFLP Patterns in Authentication --- p.126 / Chapter 8.5 --- Conclusions --- p.127 / References --- p.128 / Appendix --- p.135

Cao Hui. / Publication date from spine. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1995. / Includes bibliographical references (leaves 180-194). / Acknowledgments --- p.v / Abstract --- p.vii / List of Tables --- p.xv / List of Figures --- p.xvii / Abbreviations and symbols --- p.xx / Chapter Chapter 1. --- General introduction / Chapter 1.1. --- Historical background --- p.1 / Chapter 1.2. --- Pharmacognostical development --- p.2 / Chapter 1.3. --- Importance of herb authentication --- p.3 / Chapter 1.4. --- Objective of study --- p.5 / Chapter Chapter 2. --- Literature review / Chapter 2.1. --- Botanical and taxonomic aspects --- p.9 / Chapter 2.1.1. --- Morphology --- p.9 / Chapter 2.1.2. --- Scientific names --- p.11 / Chapter 2.2. --- Chemical aspects --- p.13 / Chapter 2.3. --- Pharmacological aspects --- p.14 / Chapter 2.3.1. --- Antibacterial effect --- p.14 / Chapter 2.3.2. --- Antiphlogistic effect --- p.14 / Chapter 2.3.3. --- Antipyretic effect --- p.15 / Chapter 2.3.4. --- Effect in gastrointestinal propulsion --- p.15 / Chapter 2.3.5. --- Antineoplastic activity --- p.15 / Chapter 2.3.6. --- Hepatoprotective effect --- p.15 / Chapter 2.3.7. --- Inhibitory activity on enzymes --- p.17 / Chapter 2.3.8. --- Cardiovascular effect --- p.17 / Chapter 2.3.9. --- Acute toxicity (LD50) --- p.18 / Chapter 2.4. --- Pharmacognostical aspects --- p.18 / Chapter Chapter 3. --- Kudidan in Ben-cao literature / Chapter 3.1. --- Introduction --- p.23 / Chapter 3.2. --- Name evolution --- p.23 / Chapter 3.3. --- Natural distribution --- p.24 / Chapter 3.4. --- Characteristics --- p.25 / Chapter 3.5. --- Substitutions investigation --- p.26 / Chapter 3.6. --- Summary --- p.29 / Chapter Chapter 4. --- Morphological differences / Chapter 4.1. --- Plant identification --- p.36 / Chapter 4.1.1. --- Introduction --- p.36 / Chapter 4.1.2. --- Collection of voucher materials --- p.36 / Chapter 4.1.3. --- Plant morphology --- p.36 / Chapter 4.2. --- Macroscopical identification --- p.46 / Chapter 4.2.1. --- Introduction --- p.46 / Chapter 4.2.2. --- Materials and methods --- p.46 / Chapter 4.2.2.1. --- Commercial samples --- p.46 / Chapter 4.2.2.2. --- Macroscopical characteristics --- p.46 / Chapter 4.2.3. --- Results --- p.49 / Chapter Chapter 5. --- Histological identification / Chapter 5.1. --- Introduction --- p.58 / Chapter 5.2. --- Materials and methods --- p.59 / Chapter 5.2.1. --- Commercial samples --- p.59 / Chapter 5.2.1.1. --- Kudidan --- p.59 / Chapter 5.2.1.2. --- Pugongying --- p.59 / Chapter 5.2.1.3. --- Substitutes --- p.59 / Chapter 5.2.2. --- Authentic plant materials for comparison --- p.60 / Chapter 5.2.3. --- Methods --- p.60 / Chapter 5.2.3.1. --- Paraffin method --- p.60 / Chapter 5.2.3.2. --- Light microscopy --- p.62 / Chapter 5.2.3.3. --- Quantitative microscopy --- p.63 / Chapter 5.2.3.4. --- Scanning electron microscopy --- p.64 / Chapter 5.3. --- Results --- p.64 / Chapter 5.3.1. --- Microscopical characters of comparative plants --- p.64 / Chapter 5.3.2. --- Internal structures of herbs --- p.83 / Chapter 5.4. --- Discussion --- p.83 / Chapter Chapter 6. --- Chemical analysis / Chapter 6.1. --- Introduction --- p.99 / Chapter 6.2. --- Materials and methods --- p.100 / Chapter 6.2.1. --- Authentic samples --- p.100 / Chapter 6.2.2. --- Commercial samples --- p.100 / Chapter 6.2.3. --- Methods --- p.100 / Chapter 6.2.3.1. --- Isolation and characterization of standard substances --- p.100 / Chapter 6.2.3.2. --- Extraction of plant materials --- p.102 / Chapter 6.2.3.3. --- Thin layer chromatography --- p.102 / Chapter 6.3. --- Results and discussion --- p.104 / Chapter 6.3.1. --- TLC synopsis --- p.104 / Chapter 6.3.2. --- TLC analysis --- p.105 / Chapter 6.3.2.1. --- Qualitative evaluation of authentic plants --- p.105 / Chapter 6.3.2.2. --- Qualitative evaluation of commercial samples --- p.107 / Chapter 6.4. --- Summary --- p.107 / Chapter Chapter 7. --- Molecular fingerprinting / Chapter 7.1. --- Introduction --- p.115 / Chapter 7.2. --- Materials and methods --- p.120 / Chapter 7.2.1. --- Plant materials --- p.121 / Chapter 7.2.2. --- Herbal materials --- p.121 / Chapter 7.2.3. --- Total genomic DNA preparation --- p.121 / Chapter 7.2.3.1. --- CsCl/EtBr gradient method --- p.121 / Chapter 7.2.3.2. --- CTAB/CsCl gradient method --- p.123 / Chapter 7.2.3.3. --- CTAB miniprep method --- p.124 / Chapter 7.2.4. --- Qualitative analysis of genomic DNA --- p.125 / Chapter 7.2.5. --- Quantitative analysis of genomic DNA --- p.126 / Chapter 7.2.6. --- Genomic DNA fingerprinting --- p.126 / Chapter 7.2.6.1. --- DNA amplification --- p.126 / Chapter 7.2.6.1.1. --- AP-PCR --- p.127 / Chapter 7.2.1.1.2. --- RAPD --- p.128 / Chapter 7.2.6.2. --- Data analysis --- p.129 / Chapter 7.3. --- Results --- p.129 / Chapter 7.3.1. --- Studies on extraction of genomic DNA --- p.129 / Chapter 7.3.2. --- Genomic DNA fingerprinting by AP-PCR --- p.130 / Chapter 7.3.3. --- Genomic DNA fingerprinting by RAPD --- p.131 / Chapter 7.4. --- Discussion --- p.131 / Chapter 7.4.1. --- DNA extraction --- p.132 / Chapter 7.4.2. --- DNA fingerprinting of Kudidan --- p.136 / Chapter 7.4.3. --- Phylogenetic relationship between two genera Elephantopus and Pseudo-elephantopus of by DNA fingerprinting --- p.141 / Chapter Chapter 8. --- General summary and conclusion / Chapter 8.1. --- General summary --- p.165 / Chapter 8.1.1. --- Ben-cao investigation --- p.166 / Chapter 8.1.2. --- Investigation of commercial samples --- p.166 / Chapter 8.1.3. --- Histological characteristics --- p.167 / Chapter 8.1.4. --- Chemical analysis --- p.168 / Chapter 8.1.5. --- DNA fingerprinting --- p.168 / Chapter 8.2. --- Conclusion --- p.169 / Appendices / Chapter A) --- Solutions --- p.171 / Chapter B) --- Chinese characters cited in this Thesis --- p.173 / Chapter a) --- Herbal names --- p.173 / Chapter b) --- Book names --- p.175 / Chapter c) --- Personal names --- p.176 / Chapter d) --- Place names --- p.177 / Chapter e) --- Miscellaneous names --- p.179 / Bibliography --- p.180

Chan, Chun Kit. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 147-161). / Abstracts in English and Chinese. / Chapter Chaper 1 --- Introduction --- p.1 / Chapter 1.1 --- General introduction to cerebral stroke --- p.1 / Chapter 1.2 --- Different types of cerebral stroke --- p.2 / Chapter 1.3 --- Statistics --- p.3 / Chapter 1.4 --- Symptoms of cerebral stroke --- p.4 / Chapter 1.5 --- Complications of cerebral stroke --- p.5 / Chapter 1.6 --- Risks and preventions of cerebral stroke --- p.6 / Chapter 1.7 --- Cerebral stroke treatment --- p.8 / Chapter 1.8 --- Post stroke rehabilitation --- p.11 / Chapter 1.9 --- Mechanisms of stroke --- p.15 / Chapter 1.9.1 --- Energy production failure and loss of ionic homeostasis --- p.15 / Chapter 1.9.2 --- Excitotoxicity --- p.16 / Chapter 1.9.3 --- Calcium ions mediated toxicity --- p.17 / Chapter 1.9.4 --- Passive neuronal cell death --- p.18 / Chapter 1.9.5 --- Oxidative stress --- p.19 / Chapter 1.9.6 --- Inflammation --- p.22 / Chapter 1.9.7 --- Apoptosis --- p.25 / Chapter 1.10 --- Potential therapeutic agents for cerebral stroke --- p.24 / Chapter 1.10.1 --- Anti-oxidative enzyme and free radical scavengers --- p.24 / Chapter 1.10.2 --- Ions channel blockers and glutamate antagonists --- p.26 / Chapter 1.10.3 --- Anti-inflammatory agent --- p.28 / Chapter 1.10.4 --- Anti-apoptotic agent --- p.28 / Chapter 1.11 --- Experimental model of cerebral ischemia-reperfusion --- p.29 / Chapter 1.11.1 --- In vitro model (oxygen and glucose deprivation model) --- p.29 / Chapter 1.11.2 --- In vivo model (Middle cerebral artery occlusion) --- p.31 / Chapter 1.12 --- Traditional Chinese Medicine (TCM) --- p.32 / Chapter 1.12.1 --- General Introduction to Traditional Chinese Medicine --- p.32 / Chapter 1.12.2 --- TCM and cerebral stroke --- p.33 / Chapter 1.12.3 --- Chinese compound formula --- p.34 / Chapter 1.12.4 --- Introduction to individual herb --- p.34 / Chapter 1.12.4.1 --- Astragali Radix (Pinyin name: Huangqi) --- p.34 / Chapter 1.12.4.2 --- Rhizoma Chuanxiong (Pinyin name: Chuanxiong) --- p.35 / Chapter 1.12.4.3 --- Radix Salviae Miltorrhizae (Pinyin name: Danshen) --- p.35 / Chapter 1.12.4.4 --- Cassia Obtusifolia Linne (Pinyin name: Jue Ming Zi) --- p.36 / Chapter 1.12.4.5 --- Radix Glycyrrhizae (Pinyin name: Gancao) --- p.37 / Chapter 1.12.4.6 --- Radix Angelicae Sinensis (Pinyin name: Dongquai) --- p.37 / Chapter 1.12.4.7 --- Paeoniae Veitchii Radix (Pinyin name: Chi Shao) --- p.38 / Chapter 1.12.5 --- Salvianolic acid B --- p.39 / Chapter 1.13 --- Aim of study --- p.40 / Chapter Chapter 2 --- Materials and Methods --- p.41 / Chapter 2.1 --- Materials --- p.41 / Chapter 2.1.1 --- Drug --- p.41 / Chapter 2.1.1.1 --- Herbal Medicine --- p.41 / Chapter 2.1.1.2 --- Herbal extraction of PSR --- p.42 / Chapter 2.1.1.3 --- Herbal extraction of individual herb --- p.43 / Chapter 2.1.1.4 --- Salvianolic acid B --- p.43 / Chapter 2.1.2 --- Chemical --- p.44 / Chapter 2.1.3 --- Animal --- p.48 / Chapter 2.2 --- Methods --- p.49 / Chapter 2.2.1 --- (AAPH)- induced erythrocyte hemolysis --- p.49 / Chapter 2.2.2 --- Cell Culture study --- p.51 / Chapter 2.2.2.1 --- Cell Line --- p.51 / Chapter 2.2.2.2 --- Cell differentiation --- p.52 / Chapter 2.2.2.3 --- In vitro model of ischemia - Oxygen glucose deprivation (OGD) experiment --- p.53 / Chapter 2.2.2.4 --- Cell viability assay --- p.54 / Chapter 2.2.3 --- In vivo Study --- p.54 / Chapter 2.2.3.1 --- Cerebral blood flow (CBF) measurement --- p.54 / Chapter 2.2.3.2 --- In vivo transient focal cerebral ischemia model - Middle cerebral artery occlusion (MCAo) --- p.55 / Chapter 2.2.3.3 --- Administration of PSR --- p.57 / Chapter 2.2.3.4 --- Administration of salvianolic acid B (SAB) --- p.59 / Chapter 2.2.3.5 --- Measurement of brain infarct volume --- p.60 / Chapter 2.2.3.6 --- In vivo anti-oxidative enzyme activity determination in the brain --- p.61 / Chapter 2.2.3.6.1 --- Brain tissue preparation --- p.61 / Chapter 2.2.3.6.2 --- Tissue homogenization and protein extraction --- p.61 / Chapter 2.2.3.6.3 --- Protein concentration determination --- p.63 / Chapter 2.2.3.6.4 --- Catalase activity determination in the brain --- p.63 / Chapter 2.2.3.6.5 --- Glutathione Peroxidase (GPx) activity determination in the brain --- p.64 / Chapter 2.2.3.6.6 --- The Superoxide Dismutase (SOD) activity determination in the brain --- p.65 / Chapter 2.2.3.7 --- Behavioral Evaluation --- p.66 / Chapter 2.2.3.7.1 --- Neurological behavioural test --- p.66 / Chapter 2.2.3.7.2 --- Shuttle box escape experiment --- p.67 / Chapter 2.3 --- Statistical analyses --- p.71 / Chapter Chapter 3 --- Results --- p.72 / Chapter 3.1 --- In vitro model of ischemia - Oxygen glucose and deprivation (OGD) experiment --- p.72 / Chapter 3.2 --- AAPH assay of PSR --- p.75 / Chapter 3.3 --- AAPH assay of individual herb --- p.77 / Chapter 3.4 --- Brain slices after middle cerebral artery occlusion (MCAo) experiment --- p.81 / Chapter 3.5 --- Brain infarct volume of single dose protocol --- p.83 / Chapter 3.6 --- Neurological behavioural test of single dose protocol --- p.85 / Chapter 3.7 --- Brain infarct volume of double doses protocol --- p.87 / Chapter 3.8 --- Neurological behavioural test of double doses protocol --- p.89 / Chapter 3.9 --- Determination of superoxide dismutase (SOD) activity in the brain --- p.91 / Chapter 3.10 --- Determination of glutathione peroxidase (GPx) activity in the brain --- p.93 / Chapter 3.11 --- Determination of catalase activity in the brain --- p.95 / Chapter 3.12 --- Brain infarction volume of Salvianolic acid B (SAB) treatment --- p.98 / Chapter 3.13 --- Neurological behavioural test of SAB treatment --- p.100 / Chapter 3.14 --- Shuttle box performance in training and testing series --- p.102 / Chapter 3.15 --- Change in shuttle box performance (% avoidance c.f. last day of training) in testing series --- p.104 / Chapter 3.16 --- Escape latency in testing and training series --- p.107 / Chapter 3.17 --- Change in escape latency (c.f. last day of training) in testing series --- p.109 / Chapter 3.18 --- Brain infarct volume of shuttle box escape experiment --- p.112 / Chapter 3.19 --- Neurological score in shuttle box escape experiment --- p.114 / Chapter Chapter 4 --- Discussion --- p.117 / Chapter 4.1 --- The protective effect of PSR in in vitro oxygen and glucose deprivation (OGD) on human neuroblastoma SH-SY5Y cell line --- p.117 / Chapter 4.1.1 --- OGD model and cell line --- p.117 / Chapter 4.1.2 --- Protective effect of PSR in OGD experiment --- p.118 / Chapter 4.1.3 --- Free radical scavenging property of PSR --- p.120 / Chapter 4.2 --- The protective effects of PSR in in vivo middle cerebral artery (MCAo) model --- p.121 / Chapter 4.2.1 --- The shortcomings of in vitro OGD model --- p.121 / Chapter 4.2.2 --- Development of in vivo MCAo model and TTC staining --- p.122 / Chapter 4.2.3 --- Protective effect of PSR in MCAo experiment (single dose protocol) --- p.124 / Chapter 4.2.4 --- Protective effect of PSR in MCAo experiment (double doses protocol) --- p.125 / Chapter 4.2.5 --- The effect of PSR toward neurological deficits --- p.127 / Chapter 4.2.6 --- Anti-oxidative effects of PSR in MCAo model --- p.128 / Chapter 4.3 --- The protective effects of SAB in in vivo middle cerebral artery (MCAo) model --- p.130 / Chapter 4.3.1 --- Free radical scavenging property of different herbs --- p.130 / Chapter 4.3.2 --- Selection of pure compound that used to treat stroke --- p.131 / Chapter 4.3.3 --- Protective effect of Salvianolic B in MCAo experiment --- p.132 / Chapter 4.3.4 --- The effect of SAB toward neurological deficits --- p.133 / Chapter 4.4 --- The effects of PSR and SAB on stroked rats' performance in shuttle box escape experiment --- p.134 / Chapter 4.4.1 --- Establishment of shuttle box escape experiment --- p.134 / Chapter 4.4.2 --- Effects of PSR and SAB on avoidance performance --- p.135 / Chapter 4.4.3 --- Effects of PSR and SAB on escape latency --- p.138 / Chapter 4.5 --- Assessment on the contribution of SAB to the protective effect of PSR --- p.140 / Chapter 4.6 --- Comparison of acute and chronic testing --- p.140 / Chapter 4.6.1 --- The protective effect of the drugs (Histopathological examination) --- p.140 / Chapter 4.6.2 --- The severity of motor deficit (Neurological score) --- p.141 / Chapter Chapter 5 --- Conclusion and Future prospect --- p.143 / Chapter 5.1 --- Conclusion --- p.143 / Chapter 5.2 --- Future prospect --- p.144 / References --- p.147

Chan, Hoi Ling. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 206-220). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABBREVIATIONS --- p.ii / ABSTRACT --- p.viii / 撮要 --- p.xii / PUBLICATIONS --- p.xiv / TABLE OF CONTENTS --- p.xv / Chapter CHAPTER 1: --- GENERAL INTRODUCTION / Chapter 1.1 --- Hematopoiesis & Leukemia --- p.1 / Chapter 1.1.1 --- An Overview on Hematopoiesis Development --- p.1 / Chapter 1.1.2 --- Leukemia --- p.6 / Chapter 1.1.2.1 --- General Symptoms of Leukemia --- p.7 / Chapter 1.1.2.2 --- Classification of Leukemia --- p.8 / Chapter 1.1.2.3 --- Conventional Treatment against Leukemia --- p.15 / Chapter 1.1.2.4 --- Novel Approaches --- p.19 / Chapter 1.2 --- The Chinese Medicinal Herb-Banlangen (板藍根） --- p.24 / Chapter 1.2.1 --- An Overview on Natural Indigo Compounds Derived from Banlangen --- p.24 / Chapter 1.2.2 --- Tryptanthrin --- p.29 / Chapter 1.2.2.1 --- Anti-bacterial Activity of Tryptanthrin --- p.29 / Chapter 1.2.2.2 --- Anti-tumor Activity of Tryptanthrin --- p.31 / Chapter 1.2.2.3 --- Anti-inflammatory Activity of Tryptanthrin --- p.33 / Chapter 1.2.2.4 --- Cutting Edges of Tryptanthrin as a Drug --- p.34 / Chapter 1.2.2.5 --- Metabolism of Tryptanthrin --- p.35 / Chapter 1.3 --- Aims and Scopes of This Investigation --- p.37 / Chapter CHAPTER 2: --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.39 / Chapter 2.1.1 --- Animals --- p.39 / Chapter 2.1.2 --- Cell Lines --- p.39 / Chapter 2.1.3 --- "Cell Culture Medium, Buffers and Other Reagents" --- p.41 / Chapter 2.1.4 --- Reagents for 3H-Thymidine Incorporation Assay --- p.45 / Chapter 2.1.5 --- Reagents and Buffers for Flow Cytometry --- p.46 / Chapter 2.1.6 --- Reagents for DNA Extraction --- p.49 / Chapter 2.1.7 --- Reagents for Measuring Caspase Activity --- p.50 / Chapter 2.1.8 --- Reagents for Total RNA Isolation --- p.53 / Chapter 2.1.9 --- Reagents and Buffers for Reversed Transcription-PCR --- p.54 / Chapter 2.1.10 --- Reagents and Buffers for Real Time-PCR --- p.59 / Chapter 2.1.11 --- Reagents and Buffers for Gel Electrophoresis of Nucleic Acids --- p.59 / Chapter 2.1.12 --- "Reagents, Buffers and Materials for Western Blot Analysis" --- p.61 / Chapter 2.2 --- Methods --- p.70 / Chapter 2.2.1 --- Culture of the Tumor Cell Lines --- p.70 / Chapter 2.2.2 --- "Isolation, Preparation and Culture of Mouse Peritoneal Macrophages" --- p.70 / Chapter 2.2.3 --- Determination of Cell Viability --- p.71 / Chapter 2.2.4 --- Determination of Cell Proliferation by [3H]-TdR Incorporation Assay --- p.72 / Chapter 2.2.5 --- Determination of Anti-leukemia Activity In Vivo --- p.73 / Chapter 2.2.6 --- Analysis of Cell Cycle Profile/DNA Content by Flow Cytometry --- p.74 / Chapter 2.2.7 --- Measurement of Apoptosis --- p.75 / Chapter 2.2.8 --- Determination of the Mitochondrial Membrane Potential --- p.77 / Chapter 2.2.9 --- Measurement of Caspase Activity --- p.78 / Chapter 2.2.10 --- Study of Intracellular Accumulation of Reactive Oxygen Species --- p.79 / Chapter 2.2.11 --- Gene Expression Study --- p.80 / Chapter 2.2.12 --- Protein Expression Study --- p.83 / Chapter 2.2.13 --- Measurement of Cell Differentiation --- p.87 / Chapter CHAPTER 3: --- STUDIES ON THE ANTI-PROLIFERATIVE EFFECT OF TRYPTANTHRIN AND INDIRUBIN-3'-OXIME ON MYELOID LEUKEMIA CELLS / Chapter 3.1 --- Introduction --- p.90 / Chapter 3.2 --- Results --- p.94 / Chapter 3.2.1 --- Effects of Indirubin-3'-oxime and Tryptanthrin on the Proliferation of Myeloid Leukemia Cell Lines of Human and Murine Origins In Vitro --- p.94 / Chapter 3.2.2 --- Kinetic and Reversibility Studies of the Anti-proliferative Effect of Tryptanthrin on Murine Myelomonocytic Leukemia WEHI-3B JCS Cells In Vitro --- p.108 / Chapter 3.2.3 --- Cytotoxic Effect of Tryptanthrin on Murine Myelomonocytic Leukemia WEHI-3B JCS Cells In Vitro --- p.113 / Chapter 3.2.4 --- Cytotoxicity of Tryptanthrin on Non-Cancer Cell Line and Primary Myeloid Cells In Vitro --- p.115 / Chapter 3.2.5 --- Effects of Tryptanthrin on the Cell Cycle Profile of the Murine Myelomonocytic Leukemia WEHI-3B JCS Cells In Vitro --- p.118 / Chapter 3.2.6 --- Effects of Tryptanthrin on the Expression of Cell Cycle Related Genes in Murine Myelomonocytic Leukemia WEHI-3B JCS Cells In Vitro --- p.123 / Chapter 3.2.7 --- Expression of CDK-inhibitors in Tryptanthrin-treated Murine Myeloid Leukemia WEHI-3B JCS Cells --- p.126 / Chapter 3.2.8 --- Effects of Tryptanthrin on the In Vivo Tumorigenicity of the Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.128 / Chapter 3.2.9 --- In Vivo Anti-tumor Effect of Tryptanthrin on Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.130 / Chapter 3.3 --- Discussion --- p.132 / Chapter CHAPTER 4: --- STUDIES ON THE APOPTOSIS-INDUCING EFFECT OF TRYPTANTHRIN ON MURINE MYELOMONOCYTIC LEUKEMIA WEHI-3B JCS CELLS / Chapter 4.1 --- Introduction --- p.139 / Chapter 4.2 --- Results --- p.143 / Chapter 4.2.1 --- Induction of DNA Fragmentation by Tryptanthrin in the Murine Myelomonocytic Leukemia WEHI-3B Cells In Vitro --- p.143 / Chapter 4.2.2 --- Induction of Phosphatidylserine Externalization by Tryptanthrin in Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.145 / Chapter 4.2.3 --- Change of Mitochondrial Membrane Potential of Tryptanthrin- treated Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.147 / Chapter 4.2.4 --- Induction of Caspase Activity in Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.150 / Chapter 4.2.5 --- Induction of Reactive Oxygen Species in Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.155 / Chapter 4.2.6 --- Expression of Bcl-2 Family Proteins in the Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.160 / Chapter 4.2.7 --- Effects of Tryptanthrin on the mRNA Expression of Bcl-2 Family Proteins in Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.163 / Chapter 4.2.8 --- Expression of Fas and Fas Ligand Proteins in Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.167 / Chapter 4.2.9 --- Expression of Pro-Apoptotic Protein in Tryptanthrin- treated Murine Myelomonocytic Leukemia WEHI-3B JCS cells --- p.170 / Chapter 4.2 --- Discussion --- p.173 / Chapter CHAPTER 5: --- STUDIES ON THE DIFFERENTIATION-INDUCING ABILITY OF TRYPTANTHRIN ON MURINE MYELOMONOCYTIC LEUKEMIA WEHI-3B JCS CELLS / Chapter 5.1 --- Introduction --- p.184 / Chapter 5.2 --- Results --- p.186 / Chapter 5.2.1 --- Morphological Studies on Tryptanthrin-treated Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.186 / Chapter 5.2.2 --- Effects of Tryptanthrin on the Cell Size and Granularity of the Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.189 / Chapter 5.2.3 --- Effects of Tryptanthrin on Induction of NBT-reducing Activity in the Murine Myelomonocytic Leukemia WEHI-3B JCS Cells --- p.191 / Chapter 5.3 --- Discussion --- p.195 / Chapter CHAPTER 6: --- CONCLUSIONS AND FUTURE PERSPECTIVES --- p.198 / REFERENCES --- p.206

[Truncated abstract] Traditional medicines are used by millions of people throughout the world as their primary source of medical care. A range of materials are in used traditional medicines including plant and animal parts. Even though the traditional medicine trade is estimated to be worth sixty billion dollars annually the trade remains largely unregulated. Unscrupulous practices by vendors to increase their profit margins such as substituting and adulterating expensive material with cheaper varieties go unchecked. This can be dangerous to consumers because some substitutions involve poisonous material. Also, animal parts from endangered species can find their way into traditional medicines, therefore there needs to be a way to identify them in traditional medicines to prosecute poachers. The traditional techniques used for the identification of material used in Traditional Chinese Medicine (TCM) include, morphological, histological, chemical and immunological analysis. However, these techniques have their limitations. This makes applying multiple techniques essential to provide thorough authentication of the material. DNA profiling provides a technique well suited to analysing material used in TCM. DNA profiling is advantageous over other techniques used to authenticate material used in TCM because it requires only a small sample amount, can determine the cultivator, be used on all forms of TCM and potentially distinguish the components of mixtures. ... Therefore, profiles of different species/individual are different and species? can be distinguished. Commercially sold traditional medicines are processed which is likely to degrade the DNA of the sample making extraction and amplification difficult. Here an organic Phenol:Chloroform extraction technique extracted DNA from commercial dried root samples. The extracted DNA was amplifiable using RAPD primers. The RAPD primers used here produced enough polymorphic bands to distinguish different plant species. They were used to distinguish commercial samples that were sold as three different species within the Panax genus, Panax ginseng, Panax quinquefolium and Panax notoginseng and genetically unrelated plant material; Potato and Eleutherococcus senticosus. Liquid samples and mixtures were also profiled with the RAPD primers to determine whether the RAPD primers provide enough distinguishing ability to analyse these forms of TCM. DNA was extracted from the liquid samples, one a ginseng drink and the other an ginseng extractum. However, there was no reliability in the production of PCR products. The analysis of the mixture samples found that not enough polymorphic bands were produced by the RAPD primers used here to identify Panax species within mixtures of two Panax species. While when P. ginseng was mixed with a genetically unrelated sample there was enough polymorphism to differentiate the two samples in the mixture. The results of this research show that RAPD analysis provides a simple and inexpensive technique to begin analysis of materials used in TCM. Using RAPD analysis it is possible to distinguish Panax plant species from each other. However, the RAPD primers used here did not provide enough reproducibility or polymorphism to analyse liquid and mixtures of Panax species plants.