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Gene expression profiling of cardinal ligament in Hong Kong Chinese women with uterine prolapse.

Liu Yuet Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 165-191). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Abstract --- p.iii / Abbreviations --- p.vi / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Incidences and Prevalence --- p.2 / Chapter 1.2 --- Anatomy of Uterus and its Support Mechanism --- p.3 / Chapter 1.3 --- Pathophysiology of Uterine Prolapse --- p.5 / Chapter 1.4 --- Classification of Uterine Prolapse --- p.6 / Chapter 1.5 --- Etiology of Uterine Prolapse --- p.7 / Chapter 1.6 --- Treatment of Uterine Prolapse --- p.12 / Chapter 1.6.1 --- Conservative Treatment --- p.12 / Chapter 1.6.2 --- Surgical Treatment --- p.13 / Chapter 1.7 --- Molecular Basis of Uterine Prolapse --- p.14 / Chapter 1.7.1 --- Collagen Metabolism --- p.15 / Chapter 1.7.2 --- Extracellular Matrix Metabolism --- p.16 / Chapter 1.7.3 --- Advanced Glycation End-products --- p.18 / Chapter 1.7.4 --- Estrogen and Estrogen Receptors --- p.19 / Chapter 1.8 --- Gene Expression Profiling of Uterine Prolapse --- p.22 / Chapter 1.9 --- Microarray Gene Expression Profiling Analysis --- p.24 / Chapter 1.9.1 --- Types of Microarray --- p.26 / Chapter 1.9.2 --- Comparison of Oligonucleotide and cDNA Arrays --- p.31 / Chapter 1.10 --- Quantitative Real-time PCR --- p.32 / Chapter 1.10.1 --- Principle of TaqMan Real-time PCR --- p.32 / Chapter 1.10.2 --- Other Types of Real-time PCR --- p.33 / Chapter 1.11 --- Project Aims --- p.34 / Chapter 1.12 --- Significance of Study --- p.35 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.37 / Chapter 2.1 --- Materials --- p.37 / Chapter 2.1.1 --- Patients --- p.37 / Chapter 2.1.2 --- Cardinal Ligament Specimen --- p.38 / Chapter 2.2 --- Methods --- p.39 / Chapter 2.2.1 --- Homogenization of Cardinal Ligament Tissues --- p.39 / Chapter 2.2.2 --- Total RNA extraction --- p.39 / Chapter 2.2.3 --- Oligonucleotide Microarray --- p.41 / Chapter 2.2.3.1 --- Two-cycle cDNA Synthesis --- p.41 / Chapter 2.2.3.2 --- Cleanup of Double-stranded cDNA --- p.45 / Chapter 2.2.3.3 --- Synthesis of Biotin-labeled cRNA --- p.45 / Chapter 2.2.3.4 --- Cleanup and Quantification of Biotin-labeled cRNA --- p.46 / Chapter 2.2.3.5 --- Fragmenting the cRNA for Target Preparation --- p.47 / Chapter 2.2.3.6 --- Target Hybridization --- p.47 / Chapter 2.2.3.7 --- "Array Washing, Staining and Scanning" --- p.48 / Chapter 2.2.3.8 --- Statistical Analysis of Microarray Data --- p.49 / Chapter 2.2.4 --- Quantitative Real-time Polymerase Chain Reaction --- p.52 / Chapter 2.2.4.1 --- Primers and Probes --- p.52 / Chapter 2.2.4.2 --- Reverse Transcription --- p.53 / Chapter 2.2.4.3 --- Plate Setup --- p.53 / Chapter 2.2.4.4 --- Real-time PCR Reaction Mixture Setup --- p.54 / Chapter 2.2.4.5 --- Statistical Analysis of Real-time PCR Data --- p.54 / Chapter CHAPTER 3 --- RESULTS --- p.56 / Chapter 3.1 --- Microarray Gene Expression Data Analysis --- p.57 / Chapter 3.1.1 --- Unsupervised Gene Selection --- p.57 / Chapter 3.1.2 --- Supervised Gene Selection --- p.59 / Chapter 3.1.2.1 --- Gene Expression Profiles Distinguish Cardinal Ligament with Uterine Prolapse from Control and Identify Differentially Expressed Genes --- p.59 / Chapter 3.1.2.2 --- Gene Expression Profiles Distinguish Cardinal Ligament with Different Degrees of Uterine Prolapse from Control and Identify Differentially Expressed Genes --- p.72 / Chapter 3.1.2.3 --- Gene Expression Profiles Distinguish Cardinal Ligament with Third-degree Prolapse from First-degree Prolapse and Identify Differentially Expressed Genes --- p.92 / Chapter 3.2 --- Validation of Microarray Data by Quantitative Real-time PCR --- p.96 / Chapter 3.2.1 --- Fold Change of Candidate Genes --- p.97 / Chapter 3.2.2 --- Correlation Between Microarray and Quantitative Real-time PCR Results --- p.102 / Chapter CHAPTER 4 --- DISCUSSIONS --- p.103 / Chapter 4.1 --- Global Gene Expression Profiling using Oligonucleotide Microarray --- p.103 / Chapter 4.1.1 --- Advantages of using Affymetrix GeneChipR Microarray for Gene Expression Profiling --- p.103 / Chapter 4.1.2 --- Microarray analysis software --- p.105 / Chapter 4.1.2.1 --- DNA-Chip Analyzer Software --- p.105 / Chapter 4.1.2.2 --- Comparison of Statistical Methods for Analysis of A ffymetrix GeneChipRMicroarray Data --- p.108 / Chapter 4.2 --- Validation of Microarray Data --- p.111 / Chapter 4.2.1 --- Advantages of using Quantitative Real-time PCR for mRNA Quantification --- p.111 / Chapter 4.3 --- Microarray Gene Expression Data Analysis --- p.115 / Chapter 4.3.1 --- Unsupervised Gene Selection --- p.115 / Chapter 4.3.2 --- Supervised Gene Selection --- p.115 / Chapter 4.3.2.1 --- Gene Expression Profiles Distinguish Cardinal Ligament with Uterine Prolapse from Control and Identify Differentially Expressed Genes --- p.115 / Chapter 4.3.2.2 --- Gene Expression Profiles Distinguish Cardinal Ligament with Different Degrees of Uterine Prolapse from Control and Identify Differentially Expressed Genes --- p.118 / Chapter 4.3.2.3 --- Gene Expression Profiles Distinguish Cardinal Ligament with Third-degree Prolapse from First-degree Prolapse and Identify Differentially Expressed Genes --- p.120 / Chapter 4.4 --- Potential Genes for Further Studies in Uterine Prolapse --- p.120 / Chapter 4.5 --- Implications of This Study --- p.157 / Chapter 4.6 --- Limitations of This Study --- p.160 / Chapter CHAPTER 5 --- CONCLUSIONS --- p.162 / Chapter CHAPTER 6 --- FUTURE PROSPECT --- p.164 / REFERENCES --- p.165

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_325639
Date January 2006
ContributorsLiu, Yuet Man., Chinese University of Hong Kong Graduate School. Division of Obstetrics and Gynaecology.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xii, 191 leaves : ill. ; 30 cm.
CoverageChina, Hong Kong, Hong Kong
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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