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