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Molecular authentication of three traditional Chinese medicines: crocodile meat, fish air-bladder and radix stellariae.January 2007 (has links)
Cheung, Chun Wai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 111-128). / Abstracts in English and Chinese. / Acknowledgement --- p.ii / Abstract --- p.iv / 摘要 --- p.vii / Table of content --- p.ix / List of Figures --- p.xvii / List of Tables --- p.xix / Abbreviations --- p.xxi / Chapter Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Complementary and Alternative Medicine (CAM) and Traditional Chinese Medicine (TCM) --- p.1 / Chapter 1.2 --- The development of Traditional Chinese Medicine --- p.2 / Chapter 1.3 --- Quality control of Traditional Chinese Medicine --- p.3 / Chapter 1.4 --- Problems of adulteration --- p.5 / Chapter 1.4.1 --- Confusion by common names --- p.5 / Chapter 1.4.2 --- Erroneous and intentional adulteration --- p.6 / Chapter 1.5 --- Authentication of Traditional Chinese Medicine using DNA techniques --- p.7 / Chapter 1.6 --- Crocodile meat --- p.10 / Chapter 1.6.1 --- Crocodile meat as Traditional Chinese Medicine --- p.10 / Chapter 1.6.2 --- Crocodile meat as exotic meat --- p.10 / Chapter 1.6.3 --- Effects of crocodile meat on mice --- p.12 / Chapter 1.6.4 --- Adulteration of crocodile meat in Hong Kong --- p.13 / Chapter 1.6.5 --- Authentication of crocodile meat --- p.14 / Chapter 1.6.5.1 --- SCAR analysis --- p.14 / Chapter 1.6.5.2 --- 12S and 16S ribosomal DNA --- p.14 / Chapter 1.7 --- Fish air-bladder --- p.15 / Chapter 1.7.1 --- Fish air-bladder as Traditional Chinese Medicine --- p.15 / Chapter 1.7.2 --- A case study --- p.16 / Chapter 1.7.3 --- Authentication of fish air-bladder --- p.17 / Chapter 1.8 --- Radix Stellariae --- p.18 / Chapter 1.8.1 --- Stellaria dichotoma L. var. lanceolata Bge --- p.18 / Chapter 1.8.2 --- Radix Stellariae as Traditional Chinese Medicine --- p.19 / Chapter 1.8.3 --- Chemicals in Radix Stellariae and their uses --- p.19 / Chapter 1.8.4 --- Adulteration of Radix Stellariae --- p.20 / Chapter 1.8.5 --- Authentication of Radix Stellariae --- p.21 / Chapter 1.8.5.1 --- Internal Transcribed Spacers (ITS) --- p.21 / Chapter 1.8.5.2 --- trnH-psbA intergenic spacer --- p.23 / Chapter 1.9 --- Objectives --- p.25 / Chapter Chapter 2. --- Materials and Methods --- p.26 / Chapter 2.1 --- Samples used in the study --- p.26 / Chapter 2.1.1 --- Crocodile and monitor lizard samples --- p.26 / Chapter 2.1.2 --- Sequence from NCBI database --- p.26 / Chapter 2.1.3 --- Fish air-bladder samples --- p.30 / Chapter 2.1.4 --- Radix Stellariae samples and samples of related species --- p.33 / Chapter 2.1.5 --- Sequences from NCBI database --- p.33 / Chapter 2.2 --- Reagents and equipments --- p.36 / Chapter 2.2.1 --- Sample preparation and DNA extraction --- p.36 / Chapter 2.2.2 --- Polymerase Chain Reaction --- p.38 / Chapter 2.2.3 --- Agarose gel electrophoresis and Gene Clean --- p.39 / Chapter 2.2.4 --- Cloning --- p.40 / Chapter 2.2.5 --- Cycle sequencing --- p.41 / Chapter 2.3 --- Experimental procedures --- p.42 / Chapter 2.3.1 --- Sample preparation --- p.42 / Chapter 2.3.2 --- DNA extraction --- p.42 / Chapter 2.3.3 --- Polymerase Chain Reaction --- p.44 / Chapter 2.3.4 --- Agarose gel electrophoresis --- p.47 / Chapter 2.3.5 --- Gene Clean --- p.47 / Chapter 2.3.6 --- Cloning --- p.48 / Chapter 2.3.7 --- Cycle sequencing and sequence analyses --- p.51 / Chapter Chapter 3. --- Crocodile meat - Results and Discussion --- p.54 / Chapter 3.1 --- Results --- p.54 / Chapter 3.1.1 --- SCAR analysis --- p.54 / Chapter 3.1.2 --- Sequence analyses --- p.55 / Chapter 3.1.3 --- The dendrograms --- p.56 / Chapter 3.2 --- Discussion --- p.60 / Chapter 3.2.1 --- SCAR as a quick and inexpensive method for the authentication of crocodile meat --- p.60 / Chapter 3.2.2 --- DNA sequencing - A useful tool to identify the source species of the crocodile meat --- p.61 / Chapter 3.2.3 --- Adulteration of crocodile meat in Hong Kong --- p.63 / Chapter 3.2.4 --- Source species of the genuine crocodile meats and the adulterants --- p.63 / Chapter 3.2.5 --- Regulation of labeling of food in Hong Kong --- p.69 / Chapter 3.2.6 --- Source species of the lizard head and tail from AFCD --- p.69 / Chapter 3.3 --- Summary --- p.70 / Chapter Chapter 4. --- Fish air-bladders - Results and Discussion --- p.72 / Chapter 4.1 --- Results --- p.72 / Chapter 4.1.1 --- Identities of sample BH and F1 --- p.73 / Chapter 4.1.2 --- Identity of sample BS --- p.74 / Chapter 4.1.3 --- Identities of samples GD and ZG --- p.74 / Chapter 4.1.4 --- Identity of sample GG --- p.74 / Chapter 4.1.5 --- "Identities of samples HB, HT and SH" --- p.75 / Chapter 4.1.6 --- Identity of sample JL --- p.75 / Chapter 4.1.7 --- Identity of sample MS --- p.76 / Chapter 4.1.8 --- Identity of sample RE --- p.76 / Chapter 4.2 --- Discussion --- p.77 / Chapter 4.2.1 --- Sample RE was confirmed to have originated from rabbit ears --- p.77 / Chapter 4.2.2 --- Identities of the dry fish air-bladders sold in Hong Kong --- p.79 / Chapter 4.2.3 --- Identities of the fresh fish air-bladders sold in Hong Kong --- p.82 / Chapter 4.2.4 --- Limitations of the use of DNA sequences for source species identification --- p.83 / Chapter 4.2.5 --- Variation in prices of fish air-bladders --- p.87 / Chapter 4.3 --- Summary --- p.88 / Chapter Chapter 5. --- Radix Stellariae - Results and Discussion --- p.89 / Chapter 5.1 --- Results --- p.89 / Chapter 5.1.1 --- Sequence analyses --- p.90 / Chapter 5.1.2 --- The dendrograms --- p.90 / Chapter 5.2 --- Discussion --- p.97 / Chapter 5.2.1 --- Identities of the samples obtained from the market --- p.97 / Chapter 5.2.2 --- Identity of sample Sdl4 --- p.97 / Chapter 5.2.3 --- Identities of samples Sd02R and Sd04 --- p.100 / Chapter 5.2.4 --- Myosoton aquaticum in the Stellaria-Myosoton clade --- p.104 / Chapter 5.2.5 --- Medicinal uses of the substitutes of Radix Stellariae --- p.105 / Chapter 5.3 --- Summary --- p.106 / Chapter Chapter 6. --- Conclusion --- p.107 / Reference --- p.111 / Appendix 1. 12S rDNA sequences of crocodilian and Varanus species obtained from NCBI database for sequence analyses --- p.129 / Appendix 2. 16S rDNA sequences of crocodilian and Varanus species obtained from NCBI database for sequence analyses --- p.130 / "Appendix 3. ITS sequences of the species in the genera Arenaria, Myosoton, Silene, and Stellaria obtained from NCBI database for sequence analyses" --- p.131 / Appendix 4. 7rnH-psbA intergenic spacer sequences of Silene species obtained from NCBI database for sequence analyses --- p.132 / Appendix 5. Sequence alignment of 12S rRNA gene sequences of crocodile and monitor lizard samples --- p.133 / Appendix 6. Sequence alignment of 16S rRNA gene sequences of crocodile and lizard samples --- p.141 / Appendix 7. Sequence alignment of coxl sequences of fish air-bladder samples --- p.149 / Appendix 8. Sequence alignment of 12S rRNA gene sequences of fish air-bladder samples --- p.151 / Appendix 9. Sequence alignment of 16S rRNA gene sequences of fish air-bladder samples --- p.153 / Appendix 10. Sequence alignment of coxl region of Vibrio parahaemolyticus and the coxl primers --- p.155 / Appendix 11. Sequence alignment of ITS sequences of Radix Stellariae and related samples --- p.156 / Appendix 12. Sequence alignment of trnH-psbA of Radix Stellariae and related samples --- p.163 / Appendix 13. Search results of coxl sequences of the fish air-bladder samples in BOLD-IDS --- p.167 / Appendix 14. Search results of coxl sequences of the fish air-bladder samples in NCBI nucleotide BLAST --- p.168 / Appendix 15. Search results of 12S rDNA sequences of the fish air-bladder samples in NCBI nucleotide BLAST --- p.169 / Appendix 16. Search results of 16S rDNA sequences of the fish air-bladder samples in NCBI nucleotide BLAST --- p.170 / Appendix 17. Sequence similarities (%) of coxl sequences of the fish air-bladder samples --- p.171 / Appendix 18. Sequence similarities (%) of 12S rDNA sequences of the fish air-bladder samples --- p.172 / Appendix 19. Sequence similarities (%) of 16S rDNA sequences of the fish air-bladder samples --- p.173
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