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Phytochemical and biological studies of phyllanthus species: effects on hepatitis B virus. / CUHK electronic theses & dissertations collectionJanuary 2005 (has links)
A number of recent research studies have been done on different species of the plants of the genus Phyllanthus. The plants are widely distributed in most tropical and subtropical countries and have long been used for the treatment of liver diseases in China and India. / Hepatitis B virus (HBV) is a major pathogen of human viral hepatitis. It has been estimated that 350 million people are chronic carriers of HBV throughout the world. Increasing evidence indicates that persistent viral infection of the liver is associated with cirrhosis and hepatocellular carcinoma. Hepatitis B virus belongs to a family of DNA viruses called hepadnaviruses. The current treatments of HBV infection with interferon or lamivudine have several disadvantages, and there appears to be much room for improvement in terms of medical treatment. / My project research focuses on two poorly characterized Indian Phyllanthus species called Phyllanthus nanus ("PN") and Phyllanthus niruri ("PI"). In my studies, random amplified polymorphic DNA ("RAPD") technique and high performance liquid chromatography ("HPLC") fingerprinting were used to authenticate different species of Phyllanthus. Both aqueous and ethanolic extracts of PN and PI were prepared to study their cytotoxicity in hepatoma cell lines. The effect of these extracts on hepatitis B virus was also examined in the HBV-genome integrated cell lines - PLC/PRF/5 (Alexander) and HepG2 2.2.15. Microparticle enzyme immunoassay ("MEIA") and enzyme-linked immunosorbent assay were used to measure the amount of hepatitis B surface antigen ("HBsAg") and hepatitis B e antigen ("HBeAg") secretion from the cell lines. RT-PCR was used to detect the change in HBsAg mRNA's expression level in the drug-treated cell lines. Real-time PCR was also employed to examine the effect of drug treatment on the level of HBV DNA replication and the amount of virions secreted into the medium. The experimental results showed that both the aqueous and ethanolic extracts of PN and PI exerted suppressive effect on HBsAg secretion and HBsAg mRNA level. The PN and PI ethanolic extracts also showed mild suppression of viral replication in vitro. The ethanolic extract of PN seemed to be more potent in suppressing HBV than the other extracts tested. (Abstract shortened by UMI.) / Lam Wai Yip. / "June 2005." / Advisers: Mary Waye; Vincent Ooi. / Source: Dissertation Abstracts International, Volume: 67-07, Section: B, page: 3594. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 217-234). / 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.
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Phyllanthus urinaria treatment in experimental model of non-alcoholic steatohepatitis. / CUHK electronic theses & dissertations collectionJanuary 2009 (has links)
Immortalized normal hepatocytes AML-12 or primary hepatocytes were cultured in control, and the methionine and choline deficient (MCD) culture medium in the presence or absence of Phyllanthus urinaria for 24 hours. Hepatocyte triglyceride contents, release of alanine aminotransferase, lipoperoxides and reactive oxygen species production were determined in the cell culture study. Age-matched wild-type C57BL/6 and diabetes db/db mice were fed control or MCD diet for 10 days with or without Phyllanthus urinaria. The levels of Hepatic steatosis, necroinflammation, triglycerides and oxidative stress were investigated. Hepatic expression of inflammatory factors and lipid regulatory mediators were assayed. The results demonstrated that Phyllanthus urinaria reduced steatosis and alanine aminotransferase (ALT) levels in culture of hepatocytes in a dose-dependent manner. Phyllanthus urinaria protected the livers against MCD-induced hepatic fat accumulation and steatohepatitis in mice. This effect was associated with repressed levels of hepatic lipid peroxides, reduced expression of cytochrome P450 (CYP) 2e1, pro-inflammatory tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), dampened activation of inflammatory C-jun N-teuninal kinase (JNK) and nuclear factor kappaB (NF-kappaB), increased expression of lipolytic Cyp4a10 and suppressed transcriptional activity of lipogenic CCAAT/enhancer binding protein beta (C/EBPbeta). Hepatic acyl co-enzyme A oxidase (ACO) that regulated hepatic beta-oxidation of fatty acid and other lipid regulators were not affected by Phyllanthus urinaria. / Non-alcoholic steatohepatitis (NASH) results from excessive accumulation of hepatic fat (steatosis) and oxidative stress. Therefore, inhibition of fatty acid cytotoxicity and liver inflammtary change is an important goal in the treatment of NASH. Phyllanthus urinaria, a herbal medicine, has been reported to have potential anti-oxidant property. We tested the effects of Phyllanthus urinaria on nutritional steatohepatitis both in vitro and in vivo, and determined the mechanism of its action. / Our study indicated that Phyllanthus urinaria effectively prevented MCD-induced steatohepatitis. This effect were probably mediated through dampening oxidative stress, ameliorating inflammation and decreasing lipid accumulation. Phyllanthus urinaria deserves further evaluation for its potential therapeutic effect on NASH in humans. / Shen, Bo. / Adviser: Henry Ly Chan. / Source: Dissertation Abstracts International, Volume: 70-09, Section: B, page: . / Thesis submitted in: November 2008. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 128-142). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.
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DNA microarray for authentication of medicinal dendrobium species. / CUHK electronic theses & dissertations collectionJanuary 2003 (has links)
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.
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Comparative studies on the biological activities of selected Chinese medicine fungi: ganoderma species and cordyceps species : an exploration of whether the different parts of their fruiting bodies bear different properties. / CUHK electronic theses & dissertations collectionJanuary 2006 (has links)
Ganoderma lucidum and Cordyceps sinensis are medicinal fungi commonly used in Chinese medicine. The allied species of G. lucidum, G. sinense and G. tsugae as well as the allied species of C. sinensis, C. militaris are also commercially available as health supplements. The present study aimed at evaluating and comparing the biological activities of the different parts of fruiting bodies of Ganoderma species (whole fruiting body, pileus, stipe, spores and spore oil) and Cordyceps species (whole fruiting body, stroma and larva/pupa). / The extracts of C. sinensis and C. mililaris could stimulate secretion in human airway epithelial cell Calu-3, implying the potentials of these extracts to modulate the mucociliary clearance. The potencies of the stimulatory effects of the stroma of fruiting bodies showed stronger effects than the larva/pupa. All parts of C. sinensis and the stroma of C. mililaris could modulate the proliferation of human peripheral blood mononuclear cells and the different potencies of immunomodulatory effects were also observed. / The results demonstrated that the hot water extracts of G. lucidum, G. sinense and G. tsugae possessed antiproliferative effects on human breast cancer cell lines MCF-7 and MDA-MB-231. The extracts from the stipes showed stronger inhibitory activities on cancer cell proliferation than those from pilei. Furthermore, the extracts from the whole fruiting body and stipe of G. lucidum possessed strong antitumor effects on the nude mice bearing MCF-7 xenografts and the BALB/c mice bearing sarcoma S-180 allografts as well as strong immunomodulatory effects in terms of stimulating splenic lymphocyte proliferative responses. The oral administrations of spores and spore oil did not show significant inhibition on MCF-7 xenografts growth but they inhibited sarcoma allografts growth effectively. / The strong biological effects of the stipe of Ganoderma and the stroma of Cordyceps were showed for the first-time. This study sheds light on how the fruiting bodies of Ganoderma or Cordyceps should be used to achieve the most out of their pharmacological properties. This study also demonstrated the novel application of Cordyceps in promoting secretion in human airway submucosal glands, which reinforces the rationale of using this fungus for treating respiratory diseases. / Yue Gar Lee Grace. / "August 2006." / Advisers: Leung Ping Chung; Kwok Pui Fung; Bik San Clara Lau. / Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1584. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 318-340). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.
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Molecular authentication of Chinese medicinal herbs.January 1997 (has links)
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
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Transcriptome based gene discovery in Artemisia annua L.January 2009 (has links)
Qi, Yan. / Thesis submitted in: December 2008. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 63-79). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.III / ABSTRACT --- p.IV / TABLE OF CONTENTS --- p.VII / LIST OF ABBREVIATIONS --- p.XI / Chapter CHAPTER 1. --- LITERATURE REVIEW --- p.1 / Chapter 1.1 --- the Plant of Artemisia annua L --- p.1 / Chapter 1.2 --- The disease of malaria --- p.3 / Chapter 1.2.1 --- The life cycle of Plasmodium parasites --- p.4 / Chapter 1.2.2 --- The Artemisinin-based combination therapies (ACTs) for the treatment of malaria --- p.5 / Chapter 1.3 --- Artemisinin --- p.8 / Chapter 1.3.1 --- The content and distribution of artemisinin --- p.8 / Chapter 1.3.2 --- The mechanism of artemisinin action --- p.9 / Chapter 1.3.2.1 --- The proposed non-specific mechanisms of action --- p.10 / Chapter 1.3.2.2 --- The proposed parasite-specific mechanisms of action --- p.11 / Chapter 1.3.3 --- The biosynthesis of artemisnin in vivo --- p.12 / Chapter 1.3.4 --- The biosynthesis of artemisinin in vitro --- p.16 / Chapter 1.4 --- Trichomes --- p.18 / Chapter 1.4.1 --- Non-glandular trichomes --- p.19 / Chapter 1.4.2 --- Glandular trichome --- p.20 / Chapter 1.4.3 --- Trichomes of Artemisia annua L --- p.21 / Chapter 1.5 --- DNA Sequencing Methods --- p.24 / Chapter 1.5.1 --- The basic principle of pyrosequencing --- p.25 / Chapter 1.5.2 --- 454 pyrosequencing and its application --- p.27 / Chapter CHAPTER 2. --- MATERIALS AND METHODS --- p.32 / Chapter 2.1 --- Chemicals --- p.32 / Chapter 2.2 --- Plant materials --- p.32 / Chapter 2.3 --- Preparation of the cDNA sample for 454 sequencing --- p.33 / Chapter 2.3.1 --- Scanning electron microscopy --- p.33 / Chapter 2.3.2 --- Isolation of glandular trichomes --- p.34 / Chapter 2.3.3 --- cDNA synthesis and normalization --- p.34 / Chapter 2.4 --- 454-EST SEQUENCING AND PROCESSING --- p.36 / Chapter 2.5 --- Analysis of 454 sequencing data --- p.37 / Chapter 2.6 --- Establishment of regeneration system of A. annua L --- p.37 / Chapter 2.6.1 --- Shoots induction from leaf discs --- p.37 / Chapter 2.6.2 --- The sensitivity of the explants to Kanamycin --- p.38 / Chapter 2.6.3 --- Rooting of the regenerated seedlings --- p.38 / Chapter CHAPTER 3. --- RESULTS AND DISCUSSION --- p.40 / Chapter 3.1 --- Glandular trichome isolation and cDNA preparation --- p.40 / Chapter 3.1.1 --- The distribution of glandular trichomes on A. annua --- p.40 / Chapter 3.1.2 --- The isolation of glandular trichomes --- p.42 / Chapter 3.1.3 --- The preparation of ds cDNA for 454 sequencing --- p.43 / Chapter 3.2 --- Pre-process of 454 pyrosequencing data --- p.44 / Chapter 3.3 --- Functional annotation of the 454-EST data --- p.47 / Chapter 3.4 --- Comparison of two sequencing runs --- p.49 / Chapter 3.5 --- Analysis of the 454 ESTs involved in secondary metabolisms --- p.50 / Chapter 3.6 --- Selection of the candidate genes --- p.55 / Chapter 3.7 --- Establishment of regeneration system of A. annua L --- p.57 / Chapter 3.7.1 --- Shoots induction from leaf discs --- p.57 / Chapter 3.7.2 --- Roots induction from shoots --- p.57 / Chapter 3.7.3 --- Sensitivity of A. annua to Kan --- p.59 / Chapter CHAPTER 4. --- CONCLUSION --- p.61 / REFERENCES --- p.63
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Molecular authentication of Chinese herbs derived from Aristolochia.January 2008 (has links)
Lam, Hilary. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 182-191). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.I / ABSTRACT --- p.III / TABLE OF CONTENTS --- p.V / LIST OF FIGURES --- p.X / LIST OF TABLES --- p.XIX / LIST OF ABBREVIATIONS --- p.XXII / Chapter Chapter 1: --- LITERATURE REVIEW --- p.1 / Chapter 1. --- Aristolochia --- p.1 / Chapter 1.1 --- "Aristolochia, as a plant" --- p.1 / Chapter 1.2 --- The chemicals in Aristolochia --- p.1 / Chapter 1.3 --- "Aristolochia, as herbal remedies" --- p.3 / Chapter 1.4 --- The Aristolochia poisoning cases --- p.4 / Chapter 1.5 --- The mechanism of AAs --- p.6 / Chapter 1.6 --- Renaming CHN to AAN --- p.6 / Chapter 1.7 --- Banning Aristolochia herbs --- p.7 / Chapter 1.8 --- The possible cause of ANN --- p.8 / Chapter 1.8.1 --- Misuse of Chinese Medicine --- p.8 / Chapter 1.8.2 --- Substitution --- p.9 / Chapter 1.8.3 --- The complexities of the herbal nomenclature --- p.9 / Chapter 1.8.4 --- Adulteration --- p.11 / Chapter 1.9 --- Methods for authenication --- p.12 / Chapter 1.9.1 --- Traditional methods for authentication --- p.12 / Chapter 1.9.2 --- The advantage of using molecular methods --- p.13 / Chapter 1.9.2.1 --- DNA fingerprinting --- p.13 / Chapter 1.9.2.2 --- DNA sequencing --- p.15 / Chapter 1.10 --- Method selection rationale --- p.15 / Chapter 1.11 --- The need for molecular authentication of six medicinal herbs --- p.17 / Chapter 1.11.1 --- The herb Mutong --- p.17 / Chapter 1.11.1.1 --- The poisoning cases reported --- p.19 / Chapter 1.11.1.2 --- Other authentication studies of Mutong --- p.19 / Chapter 1.11.2 --- The herb Muxiang --- p.20 / Chapter 1.11.2.1 --- Chemical profile --- p.21 / Chapter 1.11.2.2 --- Other authentication studies of Muxiang --- p.21 / Chapter 1.11.3 --- The herb Baiying --- p.22 / Chapter 1.11.3.1 --- The poisoning cases reported --- p.23 / Chapter 1.11.3.2 --- Other authentication studies of Baiying --- p.24 / Chapter 1.11.4 --- The herb Fangj --- p.i 24 / Chapter 1.11.4.1 --- Chemical profile --- p.25 / Chapter 1.11.4.2 --- The poisoning cases reported --- p.26 / Chapter 1.11.5 --- The herb Madouling --- p.26 / Chapter 1.11.6 --- The herb Zhushalian --- p.27 / Chapter 1.12 --- Aristolochia specific markers --- p.28 / Chapter 1.13 --- Significance of the research --- p.29 / Chapter Chapter 2: --- OBJECTIVE --- p.30 / Chapter Chapter 3: --- MATERIALS AND METHODS --- p.31 / Chapter 3.1 --- Samples source --- p.31 / Chapter 3.2 --- Total DNA extraction --- p.39 / Chapter 3.2.1 --- Cetyltriethylammonium bromide extraction --- p.39 / Chapter 3.2.2 --- Commercial kit extraction --- p.40 / Chapter 3.3 --- DNA amplification --- p.42 / Chapter 3.4 --- DNA fingerprinting --- p.43 / Chapter 3.4.1 --- DNA concentration determination --- p.43 / Chapter 3.4.2 --- ISSR fingerprinting --- p.44 / Chapter 3.5 --- Agarose gel electrophoresis --- p.45 / Chapter 3.6 --- Purification of PCR product --- p.46 / Chapter 3.7 --- Cloning of PCR product --- p.47 / Chapter 3.7.1 --- Ligation --- p.47 / Chapter 3.7.2 --- Transformation --- p.48 / Chapter 3.7.3 --- Cell cultivation --- p.48 / Chapter 3.7.4 --- Plasmid extraction --- p.49 / Chapter 3.7.5 --- Insert confirmation --- p.49 / Chapter 3.8 --- DNA sequencing --- p.50 / Chapter 3.8.1 --- Cycle sequencing --- p.50 / Chapter 3.8.2 --- Purification of cycle sequencing product --- p.51 / Chapter 3.8.3 --- DNA analysis --- p.51 / Chapter 3.9 --- Sequence analysis --- p.52 / Chapter Chapter 4: --- AUTHENICATION OF MUTONG --- p.53 / Chapter 4.1 --- Results --- p.53 / Chapter 4.1.1 --- Sequence alignment --- p.54 / Chapter 4.1.1.1 --- trnL-trnF sequences --- p.54 / Chapter 4.1.1.2 --- psbA-trnH sequences --- p.55 / Chapter 4.1.2 --- Percentage similarity analysis --- p.64 / Chapter 4.1.3 --- Dendrogram analysis --- p.67 / Chapter 4.2 --- Discussion --- p.73 / Chapter 4.2.1 --- Evaluation of chloroplast trnL-trnF region in differentiation of Mutong --- p.73 / Chapter 4.2.2 --- Evaluation of chloroplast psbA-trnH region in differentiation of Mutong --- p.74 / Chapter 4.2.3 --- Evaluation of using DNA sequencing in differentiation of Mutong --- p.75 / Chapter 4.3 --- Conclusion --- p.77 / Chapter Chapter 5: --- AUTHENICATION OF MUXIANG --- p.78 / Chapter 5.1 --- Results --- p.78 / Chapter 5.1.1 --- Sequence alignment --- p.79 / Chapter 5.1.1.1 --- trnL-trnF sequences --- p.79 / Chapter 5.1.1.2 --- psbA-trnH sequences --- p.80 / Chapter 5.1.2 --- Percentage similarity analysis --- p.88 / Chapter 5.1.3 --- Dendrogram study --- p.91 / Chapter 5.2 --- Discussion --- p.97 / Chapter 5.2.1 --- Evaluation of chloroplast trnL-trnF region in differentiation of Muxiang --- p.97 / Chapter 5.2.2 --- Evaluation of chloroplast psbA-trnH region in differentiation of Muxiang --- p.99 / Chapter 5.3 --- Conclusion --- p.100 / Chapter Chapter 6: --- AUTHENICATION OF BAIYING --- p.102 / Chapter 6.1 --- Results --- p.102 / Chapter 6.1.1 --- Sequence alignment --- p.103 / Chapter 6.1.2 --- Percentage similarity analysis --- p.107 / Chapter 6.1.3 --- Dendrogram analysis --- p.107 / Chapter 6.2 --- Discussion --- p.109 / Chapter 6.2.1 --- Evaluation of chloroplast psbA-trnH region in differentiation of Solarium and Aristolochia --- p.109 / Chapter 6.2.2 --- Molecular authentication of Baiying --- p.112 / Chapter 6.3 --- Conclusion --- p.113 / Chapter Chapter 7: --- AUTHENICATION OF FANGJI --- p.114 / Chapter 7.1 --- Results --- p.114 / Chapter 7.1.1 --- Sequence alignment --- p.115 / Chapter 7.1.1.1 --- trnL-trnF sequence --- p.115 / Chapter 7.1.1.2 --- psbA-trnH sequence --- p.116 / Chapter 7.1.2 --- Percentage similarity analysis --- p.123 / Chapter 7.1.3 --- Dendrogram study --- p.126 / Chapter 7.2 --- Discussion --- p.132 / Chapter 7.2.1 --- Evaluation of chloroplast trnL-trnF region in differentiation of Fangji --- p.132 / Chapter 7.2.2 --- Evaluation of chloroplast psbA-trnH region in differentiation of Fangji --- p.133 / Chapter 7.3 --- Conclusion --- p.133 / Chapter Chapter 8: --- AUTHENICATION OF MADOULING --- p.135 / Chapter 8.1 --- Results --- p.135 / Chapter 8.1.1 --- Sequence alignment --- p.136 / Chapter 8.1.1.1 --- trnL-trnF sequence --- p.136 / Chapter 8.1.1.2 --- psbA-trnH sequence --- p.136 / Chapter 8.1.2 --- Percentage similarity analysis --- p.143 / Chapter 8.1.3 --- Dendrogram study --- p.146 / Chapter 8.2 --- Discussion --- p.152 / Chapter 8.2.1 --- Evaluation of chloroplast trnL-trnF region in differentiation of Madouling --- p.152 / Chapter 8.2.2 --- Evaluation of chloroplast psbA-trnH region in differentiation of Madouling --- p.153 / Chapter 8.3 --- Conclusion --- p.153 / Chapter Chapter 9: --- AUTHENICATION OF ZHUSHALIAN --- p.155 / Chapter 9.1 --- Results --- p.155 / Chapter 9.1.1 --- Sequence alignment --- p.156 / Chapter 9.1.1.1 --- trnL-trnF sequence --- p.156 / Chapter 9.1.1.2 --- psbA-trnH sequence --- p.157 / Chapter 9.1.2 --- Percentage similarity analysis --- p.157 / Chapter 9.1.3 --- Dendrogram study --- p.162 / Chapter 9.2 --- Discussion --- p.166 / Chapter 9.2.1 --- Evaluation of chloroplast trnL-trnF region in differentiation of Zhushalian --- p.166 / Chapter 9.2.2 --- Evaluation of chloroplast psbA-trnH region in differentiation of Zhushalian --- p.171 / Chapter 9.3 --- Conclusion --- p.171 / Chapter Chapter 10: --- ARISTOLOCHIA SPECIFIC MARKER --- p.172 / Chapter 10.1 --- ISSR fingerprinting --- p.172 / Chapter 10.2 --- Discussion --- p.178 / Chapter Chapter 11: --- CONCLUSION --- p.180 / BIBLIOGRAPHY --- p.182 / APPENDIX - MATERIALS PREPARATION --- p.192
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Immunomodulatory effect of CUF2 and kuan dong hua in a rat model of house dust mite-induced allergic asthma.January 2007 (has links)
Ng, Chor Fung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 130-144). / Abstracts in English and Chinese. / ABSTRACT (ENGLISH VERSION) --- p.i / ABSTRACT (CHINESE VERSION) --- p.iv / ACKNOWLEDGEMENTS --- p.vi / TABLE OF CONTENTS --- p.viii / LIST OF TABLES AND FIGURES --- p.xii / ABBREVIATIONS --- p.xiv / Chapter CHAPTER 1. --- GENERAL INTRODUCTION --- p.1 / Chapter 1.1 --- Definition of asthma --- p.1 / Chapter 1.2 --- Asthma epidemiology --- p.2 / Chapter 1.3 --- Pathogenesis of Asthma --- p.3 / Chapter 1.3.1 --- Gene-environment interaction --- p.3 / Chapter 1.3.2 --- Allergens and atopic sensitization --- p.4 / Chapter 1.3.3 --- Other environmental factors --- p.5 / Chapter 1.4 --- House dust mite (HDM) --- p.5 / Chapter 1.4.1 --- Characteristics of HDM allergens --- p.5 / Chapter 1.4.2 --- HDM and asthma --- p.6 / Chapter 1.5 --- Pathophysiology of asthma --- p.8 / Chapter 1.5.1 --- Airway inflammation --- p.8 / Chapter 1.5.1.1 --- Cellular mechanism --- p.8 / Chapter 1.5.1.2 --- Characteristics of chronic inflammation --- p.9 / Chapter 1.5.1.3 --- Inflammatory cells in airway inflammation --- p.10 / Chapter 1.5.1.3.1 --- Mast cell --- p.10 / Chapter 1.5.1.3.2 --- Macrophages --- p.11 / Chapter 1.5.1.3.3 --- T lymphocytes --- p.12 / Chapter 1.5.1.3.4 --- Eosinophils --- p.12 / Chapter 1.5.1.3.5 --- Epithelial cells --- p.13 / Chapter 1.5.1.4 --- Cytokines in asthma --- p.14 / Chapter 1.5.1.4.1 --- Inflammatory cytokines --- p.14 / Chapter 1.5.1.4.1.1 --- Interleukin-4 --- p.14 / Chapter 1.5.1.4.1.2 --- Interleukin-5 --- p.14 / Chapter 1 5.1.4.1.3 --- Interleukin-6 --- p.15 / Chapter 1.5.1.4.1.4 --- Granulocyte Monocyte Colony Stimulating Factor (GM-CSF) --- p.15 / Chapter 1.5.1.4.1.5 --- Tumor Necrosis Factor-α (TNF-α) --- p.16 / Chapter 1.5.1.4.2 --- Anti-inflammatory cytokines --- p.17 / Chapter 1.5.1.4.2.1 --- Interleukin-10 --- p.17 / Chapter 1.5.1.4.2.2 --- Interferon-γ(IFN-γ) --- p.17 / Chapter 1.5.2 --- Airway hyperresponsiveness (AHR) --- p.18 / Chapter 1.5.3 --- A irway remodeling --- p.19 / Chapter 1.6 --- Asthma therapy --- p.21 / Chapter 1.6.1 --- β2-agonists --- p.21 / Chapter 1.6.2 --- Cromolyn and nedocromil --- p.21 / Chapter 1.6.3 --- Theophylline --- p.22 / Chapter 1.6.4 --- Leukotriene modifiers --- p.22 / Chapter 1.6.5 --- Corticosteroids --- p.23 / Chapter 1.7 --- Traditional Chinese Medicine --- p.24 / Chapter 1.7.1 --- Introduction --- p.24 / Chapter 1.7.2 --- Traditional Chinese Medicine (TCM) --- p.24 / Chapter 1.7.3 --- "Chinese herbal formula, CU Formula 2 (CUF2) and Kuan Dong Hua" --- p.26 / Chapter 1.8 --- Objectives of our studies --- p.28 / Chapter CHAPTER 2. --- ESTABLISHMENT OF A HDM-INDUCED ASTHMATIC ANIMAL MODEL IN SD RATS --- p.32 / Chapter 2.1 --- Introduction --- p.32 / Chapter 2.2 --- Materials and methods --- p.33 / Chapter 2.2.1 --- Buffers and solutions --- p.33 / Chapter 2.2.2 --- Animals --- p.33 / Chapter 2.2.3 --- Preparation of aluminum hydroxide gel --- p.34 / Chapter 2.2.4 --- HDMAllergen --- p.34 / Chapter 2.2.5 --- Sensitization Procedure --- p.35 / Chapter 2.2.6 --- Intratracheal instillation challenge --- p.35 / Chapter 2.2.7 --- Bronchoalveolar lavage (BAL) and BAL Cell counting --- p.36 / Chapter 2.2.8 --- Lung Histopathological Analysis --- p.37 / Chapter 2.2.9 --- Measurement of cytokine and chemokine by Enzyme-Linked Immunosorbent Assay (ELISA) --- p.39 / Chapter 2.2.10 --- Statistical Analysis --- p.40 / Chapter 2.3 --- Results --- p.41 / Chapter 2.3.1 --- Cellular Analysis of BALF --- p.41 / Chapter 2.3.2 --- Histopathology --- p.42 / Chapter 2.3.3 --- Cytokine and chemokine --- p.43 / Chapter 2.4 --- Discussion --- p.44 / Chapter CHAPTER 3. --- IMMUNOMODULATORY EFFECT OF CUF2 AND KUAN DONG HUA IN A RAT MODEL OF HDM-INDUCED ASTHMA --- p.65 / Chapter 3.1 --- Introduction --- p.65 / Chapter 3.2 --- Materials and methods --- p.67 / Chapter 3.2.1 --- Herbal materials and extraction method --- p.67 / Chapter 3.2.2 --- "Antigen sensitization, challenge, and treatment" --- p.68 / Chapter 3.2.3 --- Bronchoalveolar lavage and cell differential counts --- p.69 / Chapter 3.2.4 --- Histological Studies --- p.69 / Chapter 3.2.5 --- Measurement of BALF cytokines and chemokines --- p.70 / Chapter 3.2.6 --- "Body weight, thymus index and spleen index" --- p.70 / Chapter 3.2.7 --- Statistical analysis --- p.70 / Chapter 3.3 --- Results --- p.71 / Chapter 3.3.1 --- Effect of herbs and DXA on total cells and eosinophils in BALF --- p.71 / Chapter 3.3.2 --- Effect of herb and DXA on lung histology --- p.72 / Chapter 3.3.3 --- Effect of herbs and DXA on cytokine and chemokine level in BALF --- p.73 / Chapter 3.3.4 --- "Effect of herb and DXA on body weight, thymus index and spleen index" --- p.75 / Chapter 3.4 --- Discussion --- p.77 / Chapter CHAPTER 4. --- IMMUNOMODULATORY EFFECT OF KUAN DONG HUA ON HUMAN MAST CELLS (HMC-1) --- p.109 / Chapter 4.1 --- Introduction --- p.109 / Chapter 4.2 --- Materials and methods --- p.110 / Chapter 4.2.1 --- Reagents --- p.110 / Chapter 4.2.2 --- Cell line and Cell Culture --- p.111 / Chapter 4.2.3 --- Herb and extraction procedure --- p.111 / Chapter 4.2.4 --- Cell Viability Assay --- p.112 / Chapter 4.2.5 --- Assay of cytokine secretion --- p.113 / Chapter 4.2.6 --- Quantitative Analysis of cytokines --- p.113 / Chapter 4.2.7 --- Bacterial endotoxin contamination --- p.114 / Chapter 4.2.8 --- Statistical analysis --- p.115 / Chapter 4.3 --- Results --- p.116 / Chapter 4.3.1 --- Effect of Kuan Dong Hua on cell viability of HMC-I --- p.116 / Chapter 4.3.2 --- Effect of Kuan Dong Hua on cytokine release from HMC-I --- p.116 / Chapter 4.3.3 --- Effect of endotoxin contamination in the extract --- p.117 / Chapter 4.4 --- Discussion --- p.118 / Chapter CHAPTER 5. --- GENERAL CONCLUSION --- p.125 / Chapter 5.1 --- Conclusion --- p.125 / Chapter 5.2 --- Limitations of this study and Future work --- p.128 / REFERENCES --- p.130 / APPENDICES --- p.145 / Appendix A. Wright-Giemsa Stain for cytospin preparations --- p.145 / Appendix B. Hematoxylin & eosin (H&E) staining --- p.145 / Appendix C. Congo Red staining --- p.146 / Appendix D. Periodic acid-Schiff (PAS) staining --- p.146
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Molecular authentication of baihuasheshecao and icefish.January 2012 (has links)
Yu, Jing. / "November 2011." / Thesis (M.Phil.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 161-172). / Abstracts in English and Chinese. / Abstract --- p.I / 摘要 --- p.III / Acknowledgements --- p.V / Contents --- p.VI / List of Figures --- p.VIII / List of Tables --- p.X / Abbreviations and Symbols --- p.XII / Chapter CHAPTER 1 --- Introduction --- p.1 / Chapter 1.1 --- Phylogenetic study of Hedyotis --- p.2 / Chapter 1.1.1 --- Rubiaceae --- p.2 / Chapter 1.1.2 --- Controversial taxonomic issues --- p.9 / Chapter 1.2 --- Traditional Chinese medicine (TCM) --- p.19 / Chapter 1.2.1 --- Introduction --- p.19 / Chapter 1.2.2 --- Baihuasheshecao --- p.20 / Chapter 1.2.3 --- Authentication of Baihuasheshecao --- p.23 / Chapter 1.3 --- Icefishes in the Hong Kong market --- p.23 / Chapter 1.3.1 --- Introduction --- p.23 / Chapter 1.4 --- Molecular approach --- p.25 / Chapter 1.4.1 --- Introduction to molecular phylogeny --- p.25 / Chapter 1.4.2 --- FINS (Forensically Informative Nucleotide Sequencing) for species identification --- p.27 / Chapter 1.4.3 --- DNA sequence markers --- p.28 / Chapter 1.5 --- Objectives --- p.38 / Chapter CHAPTER 2 --- MATERIALS AND METHODOLOGY --- p.41 / Chapter 2.1 --- Materials --- p.42 / Chapter 2.2 --- DNA extraction --- p.50 / Chapter 2.3 --- Polymerase chain reaction (PCR) method --- p.51 / Chapter 2.4 --- Gel electrophoresis --- p.54 / Chapter 2.5 --- PCR production purification --- p.54 / Chapter 2.6 --- Ligation and transformation and transformation of PCR product --- p.56 / Chapter 2.7 --- DNA sequencing and sequence analyses --- p.58 / Chapter CHAPTER 3 --- USING FORENSICALLY INFORMATIVE NUCLEOTIDE SEQUENCING (FINS) TECHNOLOGY FOR SPECIES IDENTIFICATION --- p.64 / Chapter 3.1 --- Authentication of Baihuasheshecao by FINS Analysis --- p.65 / Chapter 3.1.1 --- Authentication using FINS technology --- p.65 / Chapter 3.1.2 --- Relative effectiveness of DNA regions for FINS analysis --- p.70 / Chapter 3.1.3 --- Phylogenetic interpretation --- p.72 / Chapter 3.2 --- Authentication of Salangids (Icefishes) by FINS Analysis --- p.74 / Chapter 3.2.1 --- Analysis based on mitochondrial ribosome DNA region --- p.74 / Chapter 3.2.2 --- Analysis based on mitochondrial 16S rRNA --- p.77 / Chapter 3.2.3 --- Analysis based on combined regions --- p.79 / Chapter 3.2.4 --- Phylogenetic analysis --- p.81 / Chapter 3.2.5 --- Discussion --- p.85 / Chapter 3.3 --- Conclusions --- p.88 / Chapter CHAPTER 4 --- PHYLOGENTIC STUDY OF HEDYOTIS IN CHINA AND THEIR POSITION IN SPERMACOCEAE --- p.89 / Chapter 4.1 --- Phylogentic study of Hedyotis species in Chinese --- p.90 / Chapter 4.1.1 --- Nuclear ITS region --- p.90 / Chapter 4.1.2 --- Plastid trnL intron and trnL-V intergenic spacer region --- p.94 / Chapter 4.1.3 --- Plastid trnH-psbA intergenic spacer region --- p.98 / Chapter 4.1.4 --- Plastid rbcL region --- p.102 / Chapter 4.1.5 --- Plastid matK region --- p.106 / Chapter 4.1.7 --- Combined analysis --- p.114 / Chapter 4.2 --- The phylogenetic position of Hedyotis (species in China) in the tribe of Spermacoceae s.1 --- p.121 / Chapter 4.2.1 --- Plastid trnL - F intergenic spacer region --- p.121 / Chapter 4.2.2 --- Plastid rbcL region --- p.133 / Chapter 4.2.3 --- Plastid rps16 region --- p.141 / Chapter 4.3 --- Discussion --- p.153 / Chapter 4.3.1 --- Comparison of phylogenetic utility of the six DNA regions --- p.153 / Chapter 4.3.2 --- Diplophragma section --- p.154 / Chapter 4.3.3 --- "Hedyotis, Dimetia, Euoldendandia and Gonotheca sections" --- p.156 / Chapter 4.3.4 --- The position of Hedyotis (species in China) in Spermacoceae --- p.158 / Chapter 4.4 --- Conclusions --- p.160 / REFERENCES --- p.161 / APPENDIX --- p.173
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Bioassay development for identification of cyclooxygenase-2 inhibitors of natural origin /Ringbom, Therese. January 2002 (has links)
Diss. (sammanfattning) Uppsala : Univ., 2002. / Härtill 4 uppsatser.
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