Shum Ka Chiu. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 174-182). / Abstracts in English and Chinese. / Acknowledgement --- p.ii / Abstract --- p.iii / 摘要 --- p.vi / Table of content --- p.viii / List of figures --- p.xvi / List of tables --- p.xxii / Abbreviations --- p.xxv / Chapter Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Importance of authentication of Traditional Chinese Medicines --- p.1 / Chapter 1.1.1 --- Confusing nomenclatures --- p.1 / Chapter 1.1.2 --- Similar morphologies of different medicinal materials --- p.2 / Chapter 1.1.3 --- Toxicities of medicinal materials --- p.2 / Chapter 1.1.4 --- Conservation of natural products --- p.2 / Chapter 1.2 --- TCM listed in the Pharmacopoeia of People's Republic of China --- p.3 / Chapter 1.3 --- Overview of mis-use and intoxication of TCM --- p.4 / Chapter 1.4 --- Ordinances regulating Chinese medicines as natural products --- p.7 / Chapter 1.4.1 --- Laws governing Chinese medicine --- p.7 / Chapter 1.4.2 --- Laws governing endangered species --- p.8 / Chapter 1.5 --- Current technologies in the authentication of Traditional Chinese Medicines and their limitations --- p.9 / Chapter 1.6 --- Historical applications of Radix Aconiti --- p.12 / Chapter 1.7 --- Modern applications of Radix Aconiti --- p.16 / Chapter 1.8 --- Research on Radix Aconiti and its chemical components --- p.17 / Chapter 1.8.1 --- Chemistry --- p.17 / Chapter 1.8.2 --- Pharmacology --- p.19 / Chapter 1.8.3 --- Molecular interaction --- p.22 / Chapter 1.9 --- Brief review on the systematics and phylogeny of Aconitum --- p.23 / Chapter 1.10 --- Historical applications of Radix Aucklandiae and related materials --- p.25 / Chapter 1.11 --- Modern applications of Radix Aucklandiae and related material --- p.27 / Chapter 1.12 --- Research on Aucklandiae and related material and their chemical components --- p.28 / Chapter 1.12.1 --- Chemistry --- p.28 / Chapter 1.12.2 --- Pharmacology --- p.29 / Chapter 1.13 --- Brief review on the systematics and phylogeny of Aucklandia and related medicinal species --- p.31 / Chapter 1.14 --- Authentication by DNA sequencing --- p.33 / Chapter 1.14.1 --- Introduction --- p.33 / Chapter 1.14.2 --- Criteria of sequence markers --- p.36 / Chapter 1.14.3 --- Model used to process polymorphism in DNA sequences --- p.37 / Chapter 1.15 --- Screening for novel markers --- p.38 / Chapter 1.15.1 --- Reason for screening novel markers --- p.38 / Chapter 1.15.2 --- Basic principle --- p.39 / Chapter 1.16 --- Introduction to gas chromatography- mass spectrometry --- p.40 / Chapter 1.16.1 --- Basic principles and components of GC-MS --- p.41 / Chapter 1.16.2 --- Advantages and limitations of GC-MS --- p.42 / Chapter 1.16.3 --- Usage of GC-MS on natural product analysis --- p.43 / Chapter 1.16.4 --- Chemometric analysis --- p.44 / Chapter 1.17 --- Objectives --- p.46 / Chapter Chapter 2. --- Materials and Methods --- p.47 / Chapter 2.1 --- Plant samples --- p.47 / Chapter 2.1.1 --- Samples of Aconitum --- p.47 / Chapter 2.1.2 --- Samples of Aucklandia and related species --- p.51 / Chapter 2.2 --- DNA extraction method --- p.58 / Chapter 2.2.1 --- Reagents --- p.58 / Chapter 2.2.2 --- Methods --- p.59 / Chapter 2.3 --- Chemical extraction methods --- p.61 / Chapter 2.4 --- Chemical standard extraction and purification method --- p.62 / Chapter 2.5 --- DNA sequencing --- p.63 / Chapter 2.5.1 --- Reagents --- p.63 / Chapter 2.5.2 --- Methods --- p.65 / Chapter 2.6 --- Genomic subtraction --- p.70 / Chapter 2.7 --- Search for species-specific markers from the subtraction library --- p.74 / Chapter 2.8 --- Gas chromatography- mass spectrometry --- p.74 / Chapter 2.9 --- GC-MS chemometric analysis --- p.75 / Chapter Chapter 3. --- Authentication of Aconitum by DNA Sequencing --- p.76 / Chapter 3.1 --- Introduction --- p.76 / Chapter 3.2 --- Methods --- p.77 / Chapter 3.3 --- Results - 5S spacer --- p.77 / Chapter 3.3.1 --- Sequence information --- p.77 / Chapter 3.3.2 --- Sequence similarity --- p.78 / Chapter 3.3.3 --- Phylogram study --- p.81 / Chapter 3.4 --- Results -psbA-trnH --- p.85 / Chapter 3.4.1 --- Sequence information --- p.85 / Chapter 3.4.2 --- Sequence similarity --- p.85 / Chapter 3.4.3 --- Phylogram study --- p.87 / Chapter 3.5 --- Discussion --- p.91 / Chapter 3.5.1 --- Overview of nuclear ribosomal 5S spacer --- p.91 / Chapter 3.5.2 --- Extensive polymorphism of 5S spacer --- p.91 / Chapter 3.5.3 --- Distribution of samples in the phylograms constructed by 5S spacer --- p.93 / Chapter 3.5.4 --- Utility of 5S spacer for authentication --- p.94 / Chapter 3.5.5 --- Overview of psbA-trnH spacer --- p.94 / Chapter 3.5.6 --- Distribution of samples in the phylograms constructed by psbA-trnH spacer --- p.95 / Chapter 3.5.7 --- A distinctive region of inversion --- p.96 / Chapter 3.5.8 --- Utility of psbA-trnH for authentication --- p.97 / Chapter Chapter 4. --- Screening for Novel Markers for Authentication of Aconitum --- p.98 / Chapter 4.1 --- Introduction --- p.98 / Chapter 4.2 --- Methods --- p.99 / Chapter 4.3 --- Results - subtracted clones --- p.99 / Chapter 4.4 --- Results - SSH6 --- p.104 / Chapter 4.4.1 --- Sequence information --- p.104 / Chapter 4.4.2 --- Sequence similarity --- p.105 / Chapter 4.5 --- Results-SSH15 --- p.107 / Chapter 4.5.1 --- Sequence information --- p.107 / Chapter 4.5.2 --- Sequence similarity --- p.107 / Chapter 4.5.3 --- Phylogram study --- p.109 / Chapter 4.6 --- Results-SSH45 --- p.113 / Chapter 4.6.1 --- Sequence information --- p.113 / Chapter 4.6.2 --- Sequence similarity --- p.113 / Chapter 4.6.3 --- Phylogram study --- p.115 / Chapter 4.7 --- Discussion --- p.119 / Chapter 4.7.1 --- Utility of subtraction in screening markers --- p.119 / Chapter 4.7.2 --- SSH6 --- p.121 / Chapter 4.7.3 --- SSH15 --- p.122 / Chapter 4.7.4 --- SSH45 --- p.123 / Chapter 4.7.5 --- Hybridization in Aconitum --- p.124 / Chapter 4.7.6 --- Inferring species identities of samples from the market --- p.126 / Chapter 4.8 --- Conclusion --- p.128 / Chapter Chapter 5. --- Assessment of Aucklandia lappa and Related Species by GC-MS --- p.129 / Chapter 5.1 --- Introduction --- p.129 / Chapter 5.2 --- Methods --- p.130 / Chapter 5.3 --- Results --- p.130 / Chapter 5.3.1 --- Extraction of essential oil --- p.130 / Chapter 5.3.2 --- GC-MS analysis --- p.131 / Chapter 5.3.3 --- Peak alignment and hierarchical cluster analysis --- p.133 / Chapter 5.3.4 --- Purification of chemical markers from Aucklandia lappa --- p.148 / Chapter 5.3.5 --- Standardization of the purified chemical markers --- p.148 / Chapter 5.3.6 --- Quantitative analysis of chemical markers --- p.152 / Chapter 5.4 --- Discussion --- p.154 / Chapter 5.4.1 --- Analysis of chemical composition --- p.154 / Chapter 5.4.2 --- A comparison on chemometric methods --- p.154 / Chapter 5.4.3 --- Similarity of chemical profiles --- p.156 / Chapter 5.4.4 --- Dendrogram analysis --- p.157 / Chapter 5.4.5 --- Utility of GC-MS in authentication of A. lappa and related species --- p.159 / Chapter 5.4.6 --- Limitations --- p.159 / Chapter 5.4.7 --- Comparison with molecular data --- p.161 / Chapter 5.4.8 --- Contents of dehydrocostuslactone and costunolide --- p.163 / Chapter 5.4.9 --- Locality study --- p.164 / Chapter 5.5 --- Conclusion --- p.165 / Chapter Chapter 6. --- General Discussion --- p.167 / Chapter 6.1 --- DNA sequencing --- p.168 / Chapter 6.2 --- Genomic subtraction --- p.169 / Chapter 6.3 --- Future work on molecular authentication --- p.170 / Chapter 6.4 --- Future work on authentication of Aconitum --- p.170 / Chapter 6.5 --- Gas chromatography- mass spectrometry --- p.171 / Chapter 6.6 --- Future work on authentication by GC-MS --- p.172 / Chapter 6.7 --- Future work on authentication of Aucklandia lappa and related species … --- p.173 / References --- p.174 / Appendix A. Sequence Alignment of 5S Spacer from Aconitum Species --- p.183 / Appendix B. Sequence Alignment of psbA- trnH Spacer from Aconitum Species --- p.188 / Appendix C. Sequences of Subtracted Clones from Aconitum --- p.191 / Appendix D. Sequence Alignment of SSH6 from Aconitum Species --- p.194 / Appendix E. Sequence Alignment of SSH15 from Aconitum Species --- p.195 / Appendix F. Sequence Alignment of SSH45 from Aconitum Species --- p.200 / Appendix G. Gas Chromatograms of Essential Oil Extracts of Aucklandia lappa and Related Species --- p.202

Yip Pui Ying. / Thesis submitted in: November 2004. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 147-184). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iii / Acknowledgment --- p.v / Abbreviations --- p.vii / Table of contents --- p.viii / List of Figures --- p.xii / List of Tables --- p.xvii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1. --- The importance of characterization of Chinese medicinal materials and the development of Chinese medicine in Hong Kong --- p.1 / Chapter 1.2. --- Methods for characterization of Chinese medicinal materials --- p.5 / Chapter 1.3. --- Molecular characterization of Chinese medicinal materials --- p.8 / Chapter 1.3.1. --- DNA sequencing --- p.9 / Chapter 1.3.2. --- DNA fingerprinting --- p.14 / Chapter 1.3.3. --- Nucleic acid hybridization --- p.19 / Chapter 1.4. --- Objectives --- p.20 / Chapter Chapter 2 --- Characterization of Plant and Fungal Materials by rDNA ITS Sequence Analysis --- p.22 / Chapter 2.1. --- Introduction --- p.22 / Chapter 2.2. --- Materials and Methods --- p.22 / Chapter 2.2.1. --- Chinese medicinal materials used in this study --- p.22 / Chapter 2.2.1.1. --- Plants and fungi for interspecific ITS study --- p.22 / Chapter 2.2.1.2. --- Plant for intraspecific ITS study and locality study --- p.33 / Chapter 2.2.2. --- Extraction of total DNA --- p.35 / Chapter 2.2.3. --- PCR amplification of ITS1 and ITS2 regions of rRNA gene --- p.35 / Chapter 2.2.4. --- Purification of PCR products --- p.38 / Chapter 2.2.5. --- Cloning using pCR-Script´ёØ Amp SK(+) Cloning Kit --- p.38 / Chapter 2.2.5.1. --- Polishing --- p.38 / Chapter 2.2.5.2. --- Ligation of inserts into pCR-Script´ёØ Amp SK(+) cloning vector --- p.38 / Chapter 2.2.5.3. --- Transformation --- p.40 / Chapter 2.2.5.4. --- PCR screening of white colonies --- p.40 / Chapter 2.2.5.5. --- Purification of PCR screening products --- p.41 / Chapter 2.2.6. --- Sequencing of ITS regions --- p.41 / Chapter 2.2.6.1. --- Cycle sequencing reaction --- p.41 / Chapter 2.2.6.2. --- Purification of sequencing extension products --- p.41 / Chapter 2.2.6.3. --- Electrophoresis by genetic analyzer --- p.42 / Chapter 2.2.7. --- Sequence analysis and alignment --- p.42 / Chapter 2.3. --- Results --- p.42 / Chapter 2.3.1. --- Extraction of total DNA --- p.42 / Chapter 2.3.2. --- PCR amplification of ITS1 and ITS2 regions of rRNA gene --- p.44 / Chapter 2.3.2.1. --- Interspecific ITS study --- p.44 / Chapter 2.3.2.2. --- Intraspecific ITS study --- p.46 / Chapter 2.3.3. --- Sequence analysis and alignment --- p.47 / Chapter 2.3.3.1. --- Interspecific ITS study --- p.47 / Chapter 2.3.3.2. --- Intraspecific ITS study --- p.56 / Chapter 2.4. --- Discussions --- p.60 / Chapter 2.4.1. --- rDNA regions used for studying Chinese medicinal materials --- p.60 / Chapter 2.4.2. --- The results agreed with previously published works --- p.60 / Chapter 2.4.3. --- Explanation of interspecific results within the Ganoderma genus --- p.60 / Chapter 2.4.4. --- Implications from interspecific comparisons --- p.60 / Chapter 2.4.5. --- Implications from intraspecific comparisons --- p.61 / Chapter Chapter 3 --- .Characterization of Astragalus membranaceus by DNA Fingerprinting / Chapter 3.1 --- Introduction --- p.62 / Chapter 3.2 --- Materials and Methods --- p.62 / Chapter 3.2.1 --- Extraction of total DNA --- p.62 / Chapter 3.2.2 --- Generation and detection of DNA fingerprints by AP-PCR --- p.63 / Chapter 3.2.3 --- Analysis of DNA fingerprints --- p.63 / Chapter 3.3 --- Results --- p.63 / Chapter 3.3.1 --- Generation of DNA fingerprints by AP-PCR --- p.63 / Chapter 3.3.2 --- Fingerprint analysis --- p.69 / Chapter 3.4 --- Discussion --- p.85 / Chapter 3.4.1 --- RP-PCR has been used on Chinese medicinal materials --- p.85 / Chapter 3.4.2 --- AP-PCR used instead of RAPD --- p.85 / Chapter 3.4.3 --- Reproducibility and amount of bands --- p.86 / Chapter 3.4.4 --- Alternatives of electrophoresis process --- p.88 / Chapter 3.4.5 --- Explanation of results --- p.88 / Chapter 3.4.6 --- Distinguishing Neimengu and Shanxi samples --- p.89 / Chapter 3.4.7 --- Further studies --- p.90 / Chapter Chapter 4 --- Characterization of Plant and Fungal Materials by DNA-DNA Hybridization on Microarrays --- p.91 / Chapter 4.1 --- Introduction --- p.91 / Chapter 4.2 --- Materials and Methods --- p.92 / Chapter 4.2.1 --- Samples for microarray study --- p.92 / Chapter 4.2.2 --- Extraction of total DNA --- p.95 / Chapter 4.2.3 --- Amplification and sequencing of ITS 1 region of rRNA gene --- p.95 / Chapter 4.2.4 --- Preparation of labeled probe --- p.95 / Chapter 4.2.5 --- Amplification of ITS1 fragments --- p.97 / Chapter 4.2.6 --- Preparation of slides --- p.103 / Chapter 4.2.7 --- Hybridization and washing --- p.104 / Chapter 4.2.8 --- Scanning and data analysis --- p.105 / Chapter 4.3 --- Results --- p.105 / Chapter 4.3.1 --- DNA extraction --- p.105 / Chapter 4.3.2 --- Amplification and sequencing of ITS1 region of rRNA gene --- p.107 / Chapter 4.3.3 --- Preparation of labeled probe and amplification of ITS1 fragments… --- p.112 / Chapter 4.3.4 --- Preparation of slides --- p.112 / Chapter 4.3.5 --- Scanning and data analysis --- p.116 / Chapter 4.4 --- Discussion --- p.134 / Chapter 4.4.1 --- Implications --- p.134 / Chapter 4.4.2 --- Applying the findings --- p.134 / Chapter 4.4.3 --- Ways to maximize specificity --- p.137 / Chapter 4.4.4 --- Optimisation --- p.138 / Chapter 4.4.5 --- Microarray may be more advantageous over sequencing --- p.138 / Chapter Chapter Five --- General Discussion and Summary --- p.140 / Chapter 5.1. --- Objectives of this study --- p.140 / Chapter 5.2. --- rDNA ITS sequencing --- p.140 / Chapter 5.2.1. --- Description of the approach and summary of the results --- p.140 / Chapter 5.2.2. --- Implications from the results --- p.140 / Chapter 5.2.3. --- Advantages and limitations of DNA sequencing --- p.141 / Chapter 5.3. --- AP-PCR fingerprinting --- p.141 / Chapter 5.3.1. --- Description of the approach and summary of the results --- p.141 / Chapter 5.3.2. --- Advantages and limitations of DNA fingerprinting --- p.142 / Chapter 5.4. --- DNA-DNA hybridization on microarrays --- p.143 / Chapter 5.4.1. --- Description of the approach and summary of the results --- p.143 / Chapter 5.4.2. --- Implications from the results --- p.143 / Chapter 5.4.3. --- Advantages and limitations of DNA hybridization on microarrays. --- p.144 / Chapter 5.5. --- Overall summary --- p.144 / Chapter 5.6. --- Future studies --- p.146 / References --- p.147 / Appendix --- p.185

Lam Rosanna Yen Yen. / Thesis submitted in: September 2004. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 142-154). / Abstracts in English and Chinese. / Abstract --- p.i / Chinese Abstract --- p.iii / Acknowledgements --- p.v / Table of Contents --- p.vi / List of Abbreviations --- p.xii / List of Figures --- p.xv / List of Tables --- p.xix / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Reactive oxygen species --- p.1 / Chapter 1.1.1 --- Intracellular sources of ROS --- p.1 / Chapter 1.1.2 --- Extracellular sources of ROS --- p.2 / Chapter 1.1.3 --- Superoxide anion radicals --- p.2 / Chapter 1.1.4 --- Hydrogen peroxide --- p.3 / Chapter 1.1.5 --- Hydroxyl radicals --- p.3 / Chapter 1.1.6 --- Singlet oxygen --- p.4 / Chapter 1.1.7 --- Peroxyl radicals and peroxides --- p.4 / Chapter 1.1.8 --- Damage of cellular structures by ROS --- p.5 / Chapter 1.2 --- Antioxidative defence in the body --- p.6 / Chapter 1.2.1 --- Antioxidant proteins --- p.6 / Chapter 1.2.2 --- Antioxidant enzymes --- p.6 / Chapter 1.2.3 --- Antioxidant compounds --- p.7 / Chapter 1.2.3.1 --- Vitamin E --- p.8 / Chapter 1.2.3.2 --- Vitamin C --- p.9 / Chapter 1.2.3.3 --- Glutathione --- p.9 / Chapter 1.2.3.4 --- Urate --- p.9 / Chapter 1.2.3.4.1 --- Purine metabolism --- p.10 / Chapter 1.2.3.4.2 --- Xanthine oxidase --- p.12 / Chapter 1.2.4 --- Oxidative stress and antioxidant defence mechanisms in RBC --- p.12 / Chapter 1.2.5 --- Oxidative stress and antioxidant defence mechanisms in LDL --- p.16 / Chapter 1.3 --- Human diseases originated from pro-oxidant conditions --- p.16 / Chapter 1.3.1 --- Atherosclerosis --- p.17 / Chapter 1.3.2 --- Ischemia /reperfusion injury --- p.17 / Chapter 1.3.3 --- Glucose-6-phosphate dehydrogenase deficiency --- p.18 / Chapter 1.3.4 --- DNA mutation --- p.18 / Chapter 1.3.5 --- Other pro-oxidant state related diseases --- p.19 / Chapter 1.4 --- Hyperuricemia and gout: diseases originated from an extreme antioxidant condition --- p.19 / Chapter 1.4.1 --- Inhibition of XOD as a treatment method for hyperuricemia --- p.20 / Chapter 1.4.2 --- Relationship between ROS injury and hyperuricemia --- p.22 / Chapter 1.5 --- Antioxidants in human nutrition --- p.23 / Chapter 1.6 --- Chinese medicinal therapeutics --- p.23 / Chapter 1.6.1 --- Rhubarb --- p.25 / Chapter 1.6.2 --- Aloe --- p.26 / Chapter 1.6.3 --- Ginger --- p.27 / Chapter 1.6.4 --- Objectives of the project --- p.30 / Chapter 1.6.5 --- Strategies applied to achieve the objectives of the present project --- p.30 / Chapter Chapter 2 --- Materials and methods --- p.31 / Chapter 2.1 --- XOD inhibition assay --- p.31 / Chapter 2.1.1 --- Assay development --- p.31 / Chapter 2.1.2 --- Dose-dependent study --- p.32 / Chapter 2.1.3 --- Reversibility of the enzyme inhibition --- p.32 / Chapter 2.1.4 --- Lineweaver-Burk plots --- p.33 / Chapter 2.2 --- Lipid peroxidation inhibition assay of mouse liver microsomes --- p.34 / Chapter 2.2.1 --- Preparation of mouse liver microsomes --- p.34 / Chapter 2.2.2 --- Basis of assay --- p.34 / Chapter 2.2.3 --- Assay procedures --- p.35 / Chapter 2.3 --- AAPH-induced hemolysis inhibition assay --- p.36 / Chapter 2.3.1 --- Preparation of RBC --- p.36 / Chapter 2.3.2 --- Basis of assay --- p.36 / Chapter 2.3.3 --- Assay procedures --- p.37 / Chapter 2.4 --- Lipid peroxidation inhibition assay of RBC membrane --- p.38 / Chapter 2.4.1 --- Preparation of RBC membrane --- p.38 / Chapter 2.4.2 --- Basis of assay --- p.39 / Chapter 2.4.3 --- Assay procedures --- p.40 / Chapter 2.5 --- ATPase protection assay --- p.41 / Chapter 2.5.1 --- Preparation of RBC membrane --- p.41 / Chapter 2.5.2 --- Preparation of malachite green (MG) reagent --- p.41 / Chapter 2.5.3 --- Basis of assay --- p.41 / Chapter 2.5.4 --- Assay procedures --- p.42 / Chapter 2.5.5 --- Determination of ATPase activities --- p.43 / Chapter 2.5.6 --- Assay buffers --- p.43 / Chapter 2.6 --- Sulfhydryl group protection assay --- p.44 / Chapter 2.6.1 --- Preparation of RBC membrane --- p.44 / Chapter 2.6.2 --- Basis of assay --- p.45 / Chapter 2.6.3 --- Assay procedures --- p.45 / Chapter 2.7 --- Lipid peroxidation inhibition assay of LDL by the AAPH method --- p.46 / Chapter 2.7.1 --- Basis of assay --- p.46 / Chapter 2.7.2 --- Assay procedures --- p.46 / Chapter 2.8 --- Lipid peroxidation inhibition assay of LDL by the hemin method --- p.47 / Chapter 2.8.1 --- Basis of assay --- p.47 / Chapter 2.8.2 --- Assay procedures --- p.47 / Chapter 2.9 --- Protein assay --- p.48 / Chapter 2.10 --- Statistical analysis --- p.48 / Chapter 2.11 --- Test compounds --- p.48 / Chapter Chapter 3 --- Xanthine oxidase inhibition assay: results and discussion --- p.49 / Chapter 3.1 --- Introduction --- p.49 / Chapter 3.2 --- Results --- p.54 / Chapter 3.3 --- Discussion --- p.59 / Chapter Chapter 4 --- Lipid peroxidation inhibition in mouse liver microsomes: results and discussion --- p.64 / Chapter 4.1 --- Introduction --- p.64 / Chapter 4.2 --- Results --- p.64 / Chapter 4.3 --- Discussion --- p.69 / Chapter Chapter 5 --- Assays on protection of RBC from oxidative damage: results and discussion --- p.71 / Chapter 5.1 --- Introduction --- p.71 / Chapter 5.2 --- Results --- p.75 / Chapter 5.2.1 --- AAPH-induced hemolysis inhibition assay --- p.75 / Chapter 5.2.2 --- Lipid peroxidation inhibition assay of RBC membranes --- p.82 / Chapter 5.2.3 --- Ca2+-ATPase protection assay --- p.88 / Chapter 5.2.4 --- Na+/K+-ATPase protection assay --- p.95 / Chapter 5.2.5 --- Sulfhydryl group protection assay --- p.100 / Chapter 5.3 --- Discussion --- p.110 / Chapter 5.3.1 --- AAPH-induced hemolysis inhibition assay --- p.110 / Chapter 5.3.2 --- Lipid peroxidation inhibition assay of RBC membranes --- p.111 / Chapter 5.3.3 --- Ca2+-ATPase protection assay --- p.113 / Chapter 5.3.4 --- Na+/K+-ATPase protection assay --- p.114 / Chapter 5.3.5 --- Sulfhydryl group protection assay --- p.115 / Chapter 5.3.6 --- Chapter summary --- p.117 / Chapter Chapter 6 --- Lipid peroxidation inhibition assay of LDL: results and discussion --- p.118 / Chapter 6.1 --- Introduction --- p.118 / Chapter 6.2 --- Results --- p.118 / Chapter 6.3 --- Discussion --- p.134 / Chapter Chapter 7 --- General discussion --- p.137 / References --- p.142

Yu, Wai Lam. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 79-88). / Abstracts in English and Chinese. / Acknowledgements --- p.IV / Abstract --- p.V / 論文摘要 --- p.VII / Table of Contents --- p.IX / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Cancer --- p.1 / Chapter 1.2 --- Hepatocellular Carcinoma (HCC) --- p.2 / Chapter 1.2.1 --- Risk factors causing HCC --- p.3 / Chapter 1.2.2 --- Molecular mechanism of HCC --- p.7 / Chapter 1.2.3 --- Treatment of HCC --- p.8 / Chapter 1.3 --- Tetrandrine (Tet) - A Natural Compound Derived from Traditional Chinese Medicine (TCM) --- p.10 / Chapter 1.3.1 --- Traditional Chinese Medicine (TCM) --- p.10 / Chapter 1.3.2 --- Tetrandrine (Tet) --- p.12 / Chapter 1.4 --- Molecular View of Apoptosis --- p.14 / Chapter 1.4.1 --- Overview of apoptosis --- p.14 / Chapter 1.4.2 --- Caspase cascade --- p.15 / Chapter 1.4.3 --- Bcl-2 protein family --- p.18 / Chapter 1.4.4 --- The role of mitochondria in apoptosis --- p.20 / Chapter 1.5 --- Anti-cancer Agents Inducing Apoptosis Are New Targets --- p.22 / Chapter 1.6 --- Aim of Study --- p.26 / Chapter Chapter 2 --- Materials and Methods --- p.27 / Chapter 2.1 --- Cell Culture And Treatment --- p.27 / Chapter 2.1.1 --- Cell lines used --- p.27 / Chapter 2.1.2 --- Tetrandrine (Tet) --- p.28 / Chapter 2.1.3 --- Chemicals and reagents 2 --- p.83 / Chapter 2.1.4 --- Solution preparation --- p.29 / Chapter 2.1.5 --- Procedures --- p.30 / Chapter 2.2 --- Cell viability --- p.32 / Chapter 2.2.1 --- Chemicals and reagents . --- p.32 / Chapter 2.2.2 --- Solution preparation --- p.32 / Chapter 2.2.3 --- Procedures --- p.32 / Chapter 2.3 --- Apoptosis detection --- p.34 / Chapter 2.3.1 --- Chemicals and reagents --- p.34 / Chapter 2.3.2 --- Solution preparation --- p.35 / Chapter 2.3.3 --- Procedures --- p.36 / Chapter 2.4 --- Gene expression in tet-induced apoptotic cells --- p.39 / Chapter 2.4.1 --- Chemicals and reagents --- p.39 / Chapter 2.4.2 --- Solution preparation --- p.40 / Chapter 2.4.3 --- Procedures --- p.40 / Chapter 2.5 --- Protein expression in tet-induced apoptotic cells --- p.44 / Chapter 2.5.1 --- Chemicals and reagents --- p.44 / Chapter 2.5.2 --- Solution preparation --- p.45 / Chapter 2.5.3 --- Procedures --- p.48 / Chapter 2.6 --- Cell cycle analysis of tet-treated cells --- p.54 / Chapter 2.5.1 --- Chemicals and reagents --- p.54 / Chapter 2.5.2 --- Solution preparation --- p.54 / Chapter 2.5.3 --- Procedures --- p.54 / Chapter Chapter 3 --- Result --- p.56 / Chapter Chapter 4 --- Discussion --- p.70 / Chapter 4.1 --- Dose- and Time- Dependent Inhibitory Effects of Tet were found on HuH-7 And JHH-4 Cell Lines --- p.70 / Chapter 4.2 --- Tet Is More Selective Towards Liver Cancer Cells --- p.71 / Chapter 4.3 --- The Cell Death in HuH-7 Cells Induced by Tet is Mediated Through Apoptosis --- p.72 / Chapter 4.4 --- Hepatocellular Carcinoma (HCC)Tet Induces G1 Phase Cell Cycle Arrest as Part of Its Mechanism in Inducing Apoptosis in HuH-7 Cells --- p.73 / Chapter 4.5 --- Tet Could Probably Induce G1 Phase Cell Cycle Arrest in JHH-4 Cells --- p.75 / Chapter 4.6 --- "Tet-induced Apoptosis Involves the Intrinsic, Caspase-Dependent Pathway in Both the HuH-7 and JHH-4 Cell Lines" --- p.75 / Chapter 4.7 --- Proteins in Bcl-2 Family are Involved in the Inhibitory Mechanism of Tet --- p.77 / Reference --- p.79

Lee, Kit Lin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 95-105). / Abstracts in English and Chinese. / Abstract --- p.i / 撮要 --- p.iii / Acknowledgements --- p.v / Table of contents --- p.vii / List of Tables --- p.x / List of Figures --- p.xi / List of Abbreviations --- p.xiii / Chapter Chapter 1: --- Literature Review --- p.1 / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.1.1 --- History of Botanical Studies in Hong Kong --- p.1 / Chapter 1.1.2 --- Plant Species Named after Hong Kong --- p.2 / Chapter 1.2 --- Botanical Background of Asarum Plants --- p.5 / Chapter 1.2.1 --- Plant Species under the Family of Aristolochiaceae --- p.5 / Chapter 1.2.2 --- Herba Asari --- p.6 / Chapter 1.2.3 --- Classification of Asarum hongkongense --- p.8 / Chapter 1.2.4 --- Growing Habitat of Asarum hongkongense --- p.8 / Chapter 1.3 --- Medicinal Properties of Asari --- p.10 / Chapter 1.4 --- Chemical Constituents of Asari --- p.10 / Chapter 1.5 --- Aristolochic acid and Health Issues --- p.12 / Chapter 1.5.1 --- Aristolochic Acid Intoxication --- p.12 / Chapter 1.5.2 --- Description of Aristolochic Acid --- p.13 / Chapter 1.5.3 --- Toxicities of Aristolochic Acid --- p.13 / Chapter 1.5.4 --- Aristolochic Acid-Containing Plants --- p.15 / Chapter 1.5.5 --- Control of Aristolochic Acid-Containing Products --- p.17 / Chapter 1.5.6 --- Control of Aristolochic Acid-Containing Products in Hong Kong --- p.18 / Chapter 1.6 --- Objectives of Study --- p.19 / Chapter Chapter 2: --- Macroscopic Features of Asarum hongkongense --- p.20 / Chapter 2.1 --- Introduction --- p.20 / Chapter 2.2 --- Plant Material --- p.20 / Chapter 2.2.1 --- Asarum hongkongense --- p.20 / Chapter 2.2.2 --- Herba Asari --- p.23 / Chapter 2.3 --- Macroscopic Characteristics of Aarum hongkongense --- p.23 / Chapter 2.3.1 --- Leaf --- p.23 / Chapter 2.3.2 --- Root and Rhizome --- p.25 / Chapter 2.3.3 --- Flower --- p.27 / Chapter 2.4 --- Macroscopic Characteristics of Herba Asari heterotropoidis (Liaoxixin) --- p.31 / Chapter 2.4.1 --- Leaf --- p.31 / Chapter 2.4.2 --- Root and Rhizome --- p.31 / Chapter 2.4.3 --- Flower --- p.34 / Chapter 2.5 --- Discussion --- p.36 / Chapter Chapter 3: --- Microscopic Features of Asarum hongkongense --- p.38 / Chapter 3.1 --- Introduction --- p.38 / Chapter 3.2 --- Plant Materials --- p.39 / Chapter 3.3 --- "Chemical,Reagents and Instrumentation" --- p.39 / Chapter 3.4 --- Methods --- p.39 / Chapter 3.5 --- Microscopic Characteristics of Asarum hongkongense --- p.40 / Chapter 3.5.1 --- Transverse Section of Leaf --- p.40 / Chapter 3.5.2 --- Surface View of Leaf --- p.40 / Chapter 3.5.3 --- Transverse Section of Root --- p.43 / Chapter 3.5.4 --- Transverse Section of Rhizome --- p.43 / Chapter 3.5.5 --- Powder --- p.47 / Chapter 3.5.5.1 --- Pollens --- p.47 / Chapter 3.5.5.2 --- Vessels --- p.47 / Chapter 3.5.5.3 --- Starch Grains --- p.47 / Chapter 3.6 --- Microscopic Characteristics of Herba Asari heterotropoidis (Liaoxixin) --- p.49 / Chapter 3.6.1 --- Transverse Section of Leaf --- p.49 / Chapter 3.6.2 --- Surface View of Leaf --- p.49 / Chapter 3.6.3 --- Transverse Section of Root --- p.53 / Chapter 3.6.4 --- Transverse Section of Rhizome --- p.53 / Chapter 3.6.5 --- Powder --- p.56 / Chapter 3.6.5.1 --- Starch Grains --- p.56 / Chapter 3.6.5.2 --- Vessels --- p.56 / Chapter 3.7 --- Discussion --- p.58 / Chapter Chapter 4: --- Molecular DNA Sequencing of Asarum hongkongense --- p.61 / Chapter 4.1 --- Introduction --- p.61 / Chapter 4.2 --- Sample Preparation --- p.64 / Chapter 4.3 --- Method --- p.64 / Chapter 4.3.1 --- Extraction of Total DNA --- p.64 / Chapter 4.3.2 --- PCR Amplification of ITS1 and ITS2 Regions of rRNA Gene --- p.65 / Chapter 4.3.3 --- Purification of PCR Products --- p.65 / Chapter 4.3.4 --- Sequencing of ITS Regions --- p.66 / Chapter 4.3.4.1 --- Cycle Sequencing Reaction --- p.66 / Chapter 4.3.4.2 --- Purification of Sequencing Extension Products --- p.67 / Chapter 4.3.4.3 --- Electrophoresis by Genetic Analyzer --- p.67 / Chapter 4.3.4.4 --- Sequence Analysis and Alignment --- p.67 / Chapter 4.4 --- Results and Discussion --- p.68 / Chapter 4.4.1 --- Extraction of Total DNA --- p.68 / Chapter 4.4.2 --- PCR Amplification of ITS1 and ITS2 Regions of rRNA Gene --- p.68 / Chapter 4.4.3 --- Sequence Analyses --- p.68 / Chapter Chapter 5: --- Determination of Aristolochic Acid of Asarum hongkongense --- p.80 / Chapter 5.1 --- Introduction --- p.80 / Chapter 5.2 --- Sample Preparation --- p.81 / Chapter 5.3 --- Standard Preparation --- p.81 / Chapter 5.4 --- Experimental --- p.83 / Chapter 5.4.1 --- Chemical and Reagents --- p.83 / Chapter 5.4.2 --- Methods --- p.83 / Chapter 5.4.2.1 --- High-Performance Liquid Chromatography --- p.83 / Chapter 5.4.2.2 --- Mass Spectrometry --- p.85 / Chapter 5.4.3 --- Other Instrumentation --- p.85 / Chapter 5.5 --- Method Validation --- p.85 / Chapter 5.5.1 --- Calibration --- p.85 / Chapter 5.5.2 --- Precision --- p.87 / Chapter 5.5.3 --- Recovery Test --- p.88 / Chapter 5.5.4 --- Limit of Detection --- p.89 / Chapter 5.6 --- Results and Discussion --- p.90 / Chapter Chapter 6: --- Conclusion --- p.92 / References --- p.94

Hung Sau Ling. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 131-137). / Table of Contents --- p.1 / Acknowledgements --- p.V / Abbreviations --- p.VI / Aim of investigation --- p.IX / Abstract --- p.XI / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1 --- Cancer Chemotherapy --- p.2 / Chapter 1.2 --- Plants as sources of useful drugs --- p.4 / Chapter 1.3 --- Potent antitumor compounds found in Goniothalamus giganteus --- p.7 / Chapter 1.4 --- Brief introduction of GONIOTRIOL --- p.8 / Chapter 1.5 --- The study on the antitumor activities of the antitumor compounds --- p.9 / Chapter 1.6 --- Biochemistry study of the anticancer agents --- p.10 / Chapter Chapter 2 --- Materials and Methods --- p.18 / Chapter 2.1 --- Materials --- p.19 / Chapter 2.1.1 --- Animals --- p.19 / Chapter 2.1.2 --- "Buffers, Culture Media and Chemicals" --- p.19 / Chapter 2.1.3 --- Cell lines --- p.20 / Chapter 2.1.4 --- Dye solutions --- p.21 / Chapter 2.1.5 --- Reagents and buffers for Agarose gel --- p.21 / Chapter 2.1.6 --- Synthetic goniotriol and its derivatives --- p.21 / Chapter 2.2 --- Methods --- p.23 / Chapter 2.2.1 --- Radioactive Precursor Incorporation Assays --- p.23 / Chapter 2.2.2 --- MTT assay --- p.24 / Chapter 2.2.3 --- Neutral Red assay --- p.24 / Chapter 2.2.4 --- Isolation and preparation of cells --- p.25 / Chapter 2.2.5 --- Assay for the solvent effect --- p.25 / Chapter 2.2.6 --- Assay for the in vitro antitumor activity THC88 on different cell lines --- p.27 / Chapter 2.2.7 --- Assay of the effect of THC86 on solid sarcoma Scl80 in vivo --- p.28 / Chapter 2.2.8 --- Assay of the effect of THC86 on peritoneal Scl80 in vivo --- p.28 / Chapter 2.2.9 --- Assay of the effect of THC89 on peritoneal EAT in vivo --- p.28 / Chapter 2.2.10 --- Assay of synthetic compound (THC89 and THC87) on the mitogenic activity of spleen lymphocytes --- p.29 / Chapter 2.2.11 --- Assay of synthetic compound (THC87) on the proliferation of murine bone marrow cells from compound- treated mice --- p.30 / Chapter 2.2.12 --- "Assay of synthetic compounds (Ml, P51 and P1) on nonmalignant cell-line" --- p.31 / Chapter 2.2.13 --- Assay of antitumor activity of synthetic compound (THC86)on PU5-1.8 --- p.31 / Chapter 2.2.14 --- Assay of the cytocidal effect of THC86 --- p.32 / Chapter 2.2.15 --- "Assay on the effect of THC86 on the synthesis of DNA, RNA and protein" --- p.32 / Chapter 2.2.16 --- Direct DNA cleavage by THC86 --- p.33 / Chapter 2.2.17 --- DNA fragmentation assay / Chapter 2.2.18 --- Assay of the effect of the synthetic compound (THC86) on different growth fraction of the cells / Chapter 2.2.19 --- Mitosis Study / Chapter 2.2.20 --- Assay for the stability of the synthetic compounds / Chapter Chapter 3 --- Structure / activity relationship of the synthetic compounds --- p.36 / Chapter 3.1 --- Results --- p.37 / Chapter 3.1.1 --- In vitro antitumor activity of the synthetic compounds --- p.37 / Chapter 3.2 --- Discussion --- p.45 / Chapter Chapter 4 --- Antitumor activities of the synthetic compounds --- p.63 / Chapter 4.1 --- Results --- p.64 / Chapter 4.1.1 --- Solvent effect in the screening process --- p.64 / Chapter 4.1.2 --- The effect of the synthetic compound (THC88) on different cell lines --- p.69 / Chapter 4.1.3 --- In vivo anti-tumor activities of the synthetic compounds --- p.71 / Chapter 4.1.3a --- Effect of THC86 on solid sarcoma Sc180 in vivo --- p.71 / Chapter 4.1.3b --- Effect of THC86 on peritoneal Scl80 in vivo --- p.71 / Chapter 4.1.3c --- Effect of THC89 on peritoneal EAT in vivo --- p.72 / Chapter 4.1.4 --- Cytotoxic effect of the tested compounds on normal cells --- p.77 / Chapter 4.1.4a --- Cytotoxic effect of THC89 on normal splenocytes in vitro --- p.77 / Chapter 4.1.4b --- Effect of THC87 on the proliferation of splenocytes --- p.77 / Chapter 4.1.4c --- Effect of THC87 on the proliferation of murine bone marrow cells --- p.78 / Chapter 4.1.4d --- Cytotoxic effect on non-malignant cell-line BALB/c 3T3/A31 --- p.78 / Chapter 4.2 --- Discussion --- p.85 / Chapter Chapter 5 --- The study on the antiproliferative mechanisms of the synthetic compounds --- p.88 / Chapter 5.1 --- Results --- p.89 / Chapter 5.1.1 --- "Effect of the synthetic compounds on Cell Growth, DNA, RNA and Protein" --- p.89 / Chapter 5.1.1a --- Effect of THC86 on PU5-1.8 (macrophage-like tumor) --- p.89 / Chapter 5.1.1b --- Cytocidal effect of THC86 on EAT --- p.89 / Chapter 5.1.1c --- "Effect of the synthetic compounds on synthesis of DNA, RNA and protein" --- p.90 / Chapter 5.1.2 --- Study of the synthetic compounds on the interactions of DNA --- p.101 / Chapter 5.1.2a --- DNA cleavage assay --- p.101 / Chapter 5.1.2b --- DNA fragmentation assay --- p.101 / Chapter 5.1.3 --- Effect of the synthetic compounds on different growth fraction of the cells --- p.104 / Chapter 5.1.4 --- Mitosis study of the synthetic compounds --- p.106 / Chapter 5.1.5 --- Investigation of the stability of the synthetic compounds in culture medium --- p.112 / Chapter 5.2 --- Discussion --- p.117 / Chapter Chapter 6 --- General Discussion --- p.122 / References --- p.131

Law Sui-Lin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 99-109). / Abstracts in English and Chinese. / CONTENTS --- p.i / ABSTRACT --- p.v / 撮要 --- p.vii / ACKNOWLEDGEMENTS --- p.ix / LIST OF FIGURES & TABLES --- p.xi / ABBREVIATIONS --- p.xv / Chapter Chapter 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- The Transition of Hyperplastic to Hypertrophic Growth During Heart Development --- p.1 / Chapter 1.2 --- The Controversial Capability of Heart Regeneration --- p.3 / Chapter 1.3 --- Challenges in Treating Heart Diseases --- p.5 / Chapter 1.4 --- A New Insight Behind Traditional Chinese Medicine (TCM) for Treating Heart Diseases --- p.7 / Chapter 1.5 --- The Potential Mitogenic TCMs on Cardiomyocytes --- p.10 / Chapter 1.5.1 --- Radix Astragali --- p.11 / Chapter 1.5.2 --- Radix Ophiopogonis --- p.12 / Chapter Chapter 2 --- MATERIALS & METHODS --- p.14 / Chapter 2.1 --- Materials --- p.14 / Chapter 2.2 --- Cell Culture --- p.16 / Chapter 2.2.1 --- Primary neonatal rat cardiomyocytes cell culture --- p.16 / Chapter 2.2.1.1 --- Mayer's hemalum-eosin staining --- p.17 / Chapter 2.2.2 --- Primary rat fibroblasts cell culture --- p.18 / Chapter 2.2.3 --- H9C2 cardiac cell culture --- p.18 / Chapter 2.3 --- TCMs Preparation and Treatment --- p.19 / Chapter 2.3.1 --- Preparation of TCMs powder from aqueous extracts --- p.19 / Chapter 2.3.2 --- Preparation of culture medium with TCMs powder --- p.19 / Chapter 2.3.3 --- Pre-treatment of undifferentiated and differentiated H9C2 cardiac cells with TCMs --- p.20 / Chapter 2.3.4 --- Post-treatment of differentiated H9C2 cardiac cells with TCMs --- p.20 / Chapter 2.4 --- Assessment of DNA Synthesis and Proliferation --- p.21 / Chapter 2.4.1 --- Tritiated thymidine incorporation assay --- p.21 / Chapter 2.4.2 --- 5-Bromo-2'-deoxy-uridine (BrdU) assay --- p.22 / Chapter 2.4.3 --- Cell counting --- p.23 / Chapter 2.4.4 --- Statistical analysis --- p.23 / Chapter 2.5 --- Screening of Differentially Expressed Genes in H9C2 Cells after TCM Treatment by cDNA Microarray --- p.25 / Chapter 2.5.1 --- Total RNA extraction --- p.25 / Chapter 2.5.2 --- RNA labeling --- p.26 / Chapter 2.5.2.1 --- Synthesis of fluorescence labeled probe --- p.26 / Chapter 2.5.2.2 --- Purification of fluorescence labeled probe --- p.27 / Chapter 2.5.3 --- Microarray hybridization --- p.28 / Chapter 2.5.3.1 --- Concentration of fluorescence labeled probe --- p.28 / Chapter 2.5.3.2 --- Hybridization --- p.28 / Chapter 2.5.3.3 --- Post-hybridization treatment --- p.29 / Chapter 2.5.4 --- Data collection --- p.29 / Chapter 2.5.4.1 --- Scanning of slide --- p.29 / Chapter 2.5.4.2 --- Image processing: spots finding and quantification --- p.30 / Chapter 2.5.5 --- Data normalization and analysis --- p.30 / Chapter 2.6 --- Confirmation of Differentially Expressed Genes in H9C2 Cells after TCM Treatment by RT-PCR --- p.32 / Chapter 2.6.1 --- DNase I digestion of total RNA sample --- p.32 / Chapter 2.6.2 --- First-strand cDNA synthesis --- p.32 / Chapter 2.6.3 --- RT-PCR of the candidate genes --- p.33 / Chapter Chapter 3 --- RESULTS --- p.36 / Chapter 3.1 --- Neonatal Primary Rat Cardiomyocytes --- p.36 / Chapter 3.1.1 --- Preparation of high-purity neonatal primary rat cardiomyocytes --- p.36 / Chapter 3.1.2 --- Neonatal primary rat cardiomyocytes ceased to undergo DNA replication after 6-day in vitro culturing --- p.38 / Chapter 3.1.3 --- Both MD and HQ promoted the growth of day 1 primary rat cardiomyocytes in dose- and time-dependent manners --- p.40 / Chapter 3.1.4 --- HQ is more potent than MD in promoting the growth of day 7 primary rat cardiomyocytes --- p.43 / Chapter 3.2 --- H9C2 Cardiac cells --- p.45 / Chapter 3.2.1 --- Proliferative effect of MD and HQ on undifferentiated H9C2 cardiac cells --- p.45 / Chapter 3.2.2 --- Pre-treatment of HQ on H9C2 cardiac cells during differentiation --- p.50 / Chapter 3.2.3 --- Pre-treatment of MD and HQ on differentiated H9C2 cardiac cells --- p.52 / Chapter 3.2.4 --- Post-treatment of MD on differentiated H9C2 cardiac cells…… --- p.55 / Chapter 3.3 --- Primary Rat Fibroblasts --- p.57 / Chapter 3.3.1 --- Proliferative effect of MD and HQ on primary rat fibroblasts --- p.58 / Chapter 3.4 --- Screening of Differentially Expressed Genes in H9C2 Cells after HQ Treatment by cDNA Microarray --- p.60 / Chapter 3.4.1 --- Differentially expressed genes in undifferentiated H9C2 cardiac cells after HQ treatment --- p.60 / Chapter 3.4.2 --- Differentially expressed genes in differentiated H9C2 cardiac cells after HQ treatment --- p.66 / Chapter 3.4.3 --- Comparison of differentially expressed genes in both undifferentiated and differentiated H9C2 cardiac cells after HQ treatment --- p.72 / Chapter 3.5 --- Confirmation of Differentially Expressed Genes in H9C2 Cells after HQ Treatment by RT-PCR --- p.73 / Chapter 3.5.1 --- "Preferential up-regulation of N-G, N-G-dimethylarginine dimethylaminohydrolase mRNA expression level in undifferentiated H9C2 cardiac cells after HQ treatment " --- p.74 / Chapter 3.5.2 --- Preferential up-regulation of heme oxygenase-3 mRNA expression level in undifferentiated H9C2 cardiac cells after HQ treatment --- p.75 / Chapter 3.5.3 --- Preferential up-regulation of cyclin B mRNA expression level in differentiated H9C2 cardiac cells after HQ treatment --- p.76 / Chapter Chapter 4 --- DISCUSSION --- p.77 / Chapter 4.1 --- HQ Being a More Effective Mitogenic TCM than MD on Cardiomyocytes Exerted its Effect in Dose- and Time Dependent --- p.79 / Chapter 4.2 --- Mitogenic Effect of Both MD and HQ might Possibly Due to the Regulation of Intrinsic Factors --- p.82 / Chapter 4.3 --- HQ Rather Than MD Showed a Higher Specificity in Promoting DNA Synthesis in Cardiomyocytes --- p.83 / Chapter 4.4 --- The Differentially Expressed Genes were Supported by The Clinical Functions of HQ --- p.85 / Chapter 4.5 --- Relating the Differentially Expressed Genes with Cardiac Growth and Development --- p.87 / Chapter 4.6 --- The Hypothetic Mechanisms of Action that HQ Exerted on Cardiac Growth and Development --- p.92 / Chapter 4.7 --- Future Prospect --- p.94 / Chapter 4.7.1 --- In vivo study of HQ on the proliferation of rat cardiomyocytes from neonatal to postnatal development --- p.94 / Chapter 4.7.2 --- The study of transgenic mice carrying the target gene regulated by HQ on cardiac growth and development --- p.96 / Chapter 4.7.3 --- The determination of active component of HQ on cardiac growth and development --- p.97 / REFERENCES --- p.99 / APPENDIX --- p.110

Law Ka Yee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 214-225). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.I / ABSTRACT --- p.II / TABLE OF CONTENTS --- p.VII / LIST OF FIGURES --- p.X / LIST OF TABLES A --- p.XII / APPENDIX --- p.XIII / Chapter CHAPTER ONE --- LITERATURE REVIEW --- p.1 / Chapter 1.1 --- Chinese herbs --- p.1 / Chapter 1.1.1 --- Introduction --- p.1 / Chapter 1.1.2 --- Leigongteng --- p.2 / Chapter 1.1.2.1 --- Leigongteng and its importance --- p.2 / Chapter 1.1.2.2 --- Chemical components and pharmacological effects of Leigongteng --- p.4 / Chapter 1.1.3 --- Problems in authentication of Leigongteng --- p.5 / Chapter 1.1.3.1 --- Taxonomic problems of Tripterygium --- p.5 / Chapter 1.1.3.2 --- Confusion caused by other species --- p.7 / Chapter 1.1..3.3 --- Adulterants --- p.9 / Chapter 1.2 --- Celastraceae --- p.10 / Chapter 1.2.1 --- Introduction --- p.10 / Chapter 1.2.2 --- Controversial taxonomic issue --- p.12 / Chapter 1.2.1.1 --- Subfamilies of Celastraceae --- p.12 / Chapter 1.2.1.2 --- Subfamily Tripterygioideae --- p.13 / Chapter 1.3 --- Molecular authentication --- p.14 / Chapter 1.4 --- Molecular systematics --- p.18 / Chapter 1.4.1 --- DNA sequence markers --- p.19 / Chapter 1.4.2 --- Molecular phylogeny --- p.25 / Chapter 1.4.2.1 --- Tree-building method --- p.25 / Chapter 1.4.2.2. --- Measures of support --- p.28 / Chapter 1.5 --- Objectives --- p.29 / Chapter CHAPTER TWO --- MATERIALS AND METHODS --- p.31 / Chapter 2.1 --- Plant and herb samples --- p.31 / Chapter 2.2 --- DNA extraction --- p.41 / Chapter 2.2.1 --- Modified CTAB extraction --- p.41 / Chapter 2.2.2 --- Commercial kit extraction --- p.42 / Chapter 2.3 --- Polymerase chain reaction (PCR) condition --- p.43 / Chapter 2.4 --- DNA gel electrophoresis --- p.44 / Chapter 2.5 --- PCR product purification --- p.45 / Chapter 2.5.1 --- GEL-M´ёØ gel extraction system --- p.45 / Chapter 2.6 --- Ligation and transformation --- p.46 / Chapter 2.6.1 --- Ligation and transformation --- p.46 / Chapter 2.6.2 --- Cell cultivation --- p.47 / Chapter 2.6.3 --- Plasmid extraction --- p.47 / Chapter 2.7 --- Determination of DNA concentration --- p.49 / Chapter 2.8 --- Cycle sequencing --- p.49 / Chapter 2.9 --- Sequence analysis --- p.50 / Chapter 2.10 --- Materials preparation --- p.51 / Chapter CHAPTER THREE --- MOLECULAR AUTHENTICATION OF LEIGONGTENG --- p.54 / Chapter 3.1. --- Authentication based on internal transcribed spacer (ITS) region --- p.54 / Chapter 3.1.1 --- Sequence alignment --- p.54 / Chapter 3.1.2 --- ITS region nucleotide differences significant in authentication of Leigongteng --- p.55 / Chapter 3.1.3 --- Relationship of samples --- p.70 / Chapter 3.1.4 --- Comparison of sequences --- p.75 / Chapter 3.2 --- Authentication based on 5s-rDNA region --- p.78 / Chapter 3.2.1 --- Sequence alignment --- p.78 / Chapter 3.2.2 --- 5s-rDNA nucleotide differences significant in authentication of Leigongteng --- p.78 / Chapter 3.2.3 --- Relationship of samples --- p.88 / Chapter 3.2.4 --- Comparison of sequences --- p.90 / Chapter 3.3 --- Authentication based on psbA-trnH region --- p.93 / Chapter 3.3.1 --- Sequence alignment --- p.93 / Chapter 3.3.2 --- psbA-trnH nucleotide differences significant in authentication of Leigongteng --- p.101 / Chapter 3.3.3 --- Relationship of samples --- p.113 / Chapter 3.3.4 --- Comparison of sequences --- p.115 / Chapter 3.4 --- Authentication based on trnL-F region --- p.118 / Chapter 3.4.1 --- Sequence alignment --- p.118 / Chapter 3.4.2 --- trnL-F region nucleotide differences significant in authentication of Leigongteng --- p.121 / Chapter 3.4.3 --- Relationship of samples --- p.139 / Chapter 3.4.4 --- Comparison of sequences --- p.141 / Chapter 3.5 --- Discussion --- p.144 / Chapter 3.5.1 --- Molecular markers --- p.144 / Chapter CHAPTER FOUR --- PHYLOGENETIC STUDIES ON TRIPTERYGIUM --- p.151 / Chapter 4.1 --- Combine loci of ITS and 5s-rDNA regions --- p.152 / Chapter 4.1.1 --- Homogenity test --- p.152 / Chapter 4.1.2 --- Sequence alignment --- p.152 / Chapter 4.1.3 --- Phylogenetic study --- p.173 / Chapter 4.2 --- psbA-trnH region --- p.174 / Chapter 4.2.1 --- Sequence alignment --- p.174 / Chapter 4.3 --- Discussion --- p.177 / Chapter CHAPTER FIVE --- PHYLOGENETIC STUDIES ON TRIPTERYGIOIDEAE AND CELASTRACEAE --- p.191 / Chapter 5.1 --- ITS regions --- p.191 / Chapter 5.1.1 --- Sequence alignment --- p.191 / Chapter 5.1.2 --- Phylogenetic analysis --- p.205 / Chapter 5.2 --- Discussion --- p.206 / Chapter 5.2.1 --- Subfamily Tripterygioideae --- p.206 / Chapter 5.2.2 --- Subfamilies of Celastraceae --- p.210 / Chapter CHAPTER SIX --- CONCLUSION --- p.212 / BILBIOGRAPHY --- p.214

All the results showed that TCMs can provide a source for discovering anti-angiogenic agents for the treatment of cancer, and all these experiments in the zebrafish and mammalian models further confirmed the value of zebrafish model in anti-angiogenic drug discovery. / Angiogenesis refers to the formation of new blood capillaries from pre-existing ones, and is essential in a series of normal physiological processes such as embryonic development and pathological responses. However, persistent unregulated angiogenesis causes "angiogenic diseases" such as diabetic retinopathy, tumor growth and metastasis, rheumatoid arthritis, and inflammatory diseases. The linkage between angiogenesis, tumor growth and metastasis was first hypothesized by Dr. Judah Folkman in the 1970s, and now this controversial idea is widely accepted and the inhibition of angiogenesis, or anti-angiogenesis, is considered as a promising anticancer therapeutic strategy. Bevacizumab (Avastin RTM by Genentech Inc.), the first approved anti-angiogenic drug by U.S. FDA in 2004, is a humanized monoclonal antibody to inhibit endothelial cell proliferation and angiogenesis for the treatment of metastatic colorectal cancer, non-small cell lung cancer, advanced breast cancer, glioblastoma, metastatic renal cell cancer. / Anti-angiogenic therapy in cancer treatment has led to the development of compounds designed to control a tumor's growth by blocking its ability to develop a blood supply. The development of agents with different mechanisms of action requires powerful preclinical models for the analysis and optimization of the therapy. Some in vitro and in vivo anti-angiogenic assays are already developed, for example, Human Umbilical Vein Endothelial Cell (HUVEC) assay, Chorioallantoic Membrane assay, Matrigel plug assay et al. Zebrafish, as a relatively new model organism, is firmly established as a powerful research platform for many areas of biology and drug discovery, allowing the testing of bioactive compounds in a whole organism and in cells undergoing normal cell-cell and cell-matrix interactions. Many anti- and pro-angiogenic molecules tested in zebrafish demonstrated similar effects to those observed in humans or other mammalian models. Besides providing a powerful platform for drug screening, zebrafish model can also be used for probing biological processes, and generate insights into mechanisms. / Cancer is a generic term for a large group of diseases that can affect any part of the body, which causes a vast medical problem and is a leading cause of death worldwide nowadays. However, for many years the main methods of treating cancer have been surgery, radiotherapy and chemotherapy. Among these treatments, chemotherapy has played a major role in cancer therapy for half a century. Despite improving managements and efforts, it is not surprising that the prognosis has not greatly improved because of the limitations of current therapies, such as toxicity, inherent and acquired resistance, and metastatic spread. This calls for novel cancer therapies and new group of anticancer agents for selectively targeting cancers without or with lower toxicity to normal tissues. / Traditional Chinese medicines (TCMs) have long been recognized as a rich source for discovering drugs, and various TCMs and their components have shown anti-angiogenic properties. In this thesis study, as a continuing pursuit for elucidating the anti-angiogenic properties of TCMs, our attention is focused on those with effects of anti-inflammation, anti-rheumatoid arthritis and anti-cancer. On zebrafish screening model, three of the selected TCMs, Rheum palmatum, Alpinia oxyphylla (seeds), and Kaempferia galanga showed potential anti-angiogenic activity, indicating the existence of potent anti-angiogenic components in these herbs. The ethyl acetate fraction of R. palmatum showed strong inhibition of vessel formation in zebrafish embryos. Further testing of the anthraquinones of this herb showed three of them displayed potent anti-angiogenic activities. The most potent compound---rhein could inhibit HUVEC migration and affect the mRNA expression of vegfa, kdr, angiopoietin1/2 and tie1/2; The n-hexane and ethyl acetate fractions of A. oxyphylla and K. galangal showed anti-angiogenic potentials both in zebrafish and HUVEC assays. The n-hexane and ethyl acetate fractions of A. oxyphylla could both inhibit the proliferation, migration and tube formation processes of HUVEC. And the most potential component, trans-ethyl-p-methoxycinnamate from K. galanga, could inhibit HUVEC migration and tube formation, and reduce all gene expressions involved in angiogenesis process except for vegfa. / He, Zhiheng. / Adviser: Wei Ge. / Source: Dissertation Abstracts International, Volume: 73-02, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 87-108). / 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, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Thesis (PhD) -- University of Western Sydney, Nepean, 1998. / Bibliography : p. 164-171.