by Mo Flora. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1991. / Includes bibliographical references (leaves 168-184). / ACKNOWLEDGEMENT --- p.i / ABSTRACT --- p.ii / TABLE OF CONTENTS --- p.iv / Chapter CHAPTER ONE --- INTRODUCTION / Chapter 1.1 --- Stroke as a major disabling disease --- p.1 / Chapter 1.2 --- Classification of stroke --- p.4 / Chapter 1.3 --- Risk factors attributing to stroke --- p.15 / Chapter 1.4 --- Experimental methods to induce cerebral ischemia --- p.19 / Chapter 1.4.1 --- The establishment of animal models for stroke --- p.21 / Chapter 1.4.2 --- Gerbil as a putative model --- p.25 / Chapter 1.5 --- Mechanisms of focal ischemia damage --- p.30 / Chapter 1.6 --- Potential biochemical markers for cerebral ischemia --- p.38 / Chapter 1.7 --- Aim of investigation --- p.48 / Chapter CHAPTER TWO --- MATERIALS AND METHODS / Chapter 2.1 --- Common chemicals --- p.49 / Chapter 2.2 --- Common bench solutions --- p.52 / Chapter 2.3 --- Animals / Chapter 2.3.1 --- Gerbils --- p.52 / Chapter 2.3.2 --- Rabbit --- p.53 / Chapter 2.4 --- Establishment of an animal model / Chapter 2.4.1 --- Surgical methods for common carotid artery (CCA) ligation --- p.54 / Chapter 2.5 --- Methods to determine stroke conditions of gerbils / Chapter 2.5.1 --- Ocular fundus examination --- p.56 / Chapter 2.5.2 --- Stroke index --- p.56 / Chapter 2.5.3 --- Inclined plane method --- p.59 / Chapter 2.6 --- Preparation of gerbil brain for subsequent analysis / Chapter 2.6.1 --- Preparation of gerbil brain slices --- p.61 / Chapter 2.6.2 --- "2,3,5-triphenytetrazolium chloride (TTC) for quantitative staining of brain slices" --- p.61 / Chapter 2.6.3 --- Preparation of normal and stroke gerbil brain extract --- p.62 / Chapter 2.7 --- Polyacrylamide gel electrophoresis (PAGE) using a discontinuous buffer system / Chapter 2.7.1 --- Stock reagents --- p.63 / Chapter 2.7.2 --- Separation gel preparation --- p.65 / Chapter 2.7.3 --- Stacking gel preparation --- p.66 / Chapter 2.7.4 --- Electrophoresis conditions --- p.67 / Chapter 2.7.5 --- Staining and destaining --- p.67 / Chapter 2.8 --- Two dimensional slab gel electrophoresis / Chapter 2.8.1 --- Equipment --- p.70 / Chapter 2.8.2 --- Chemical --- p.70 / Chapter 2.8.3 --- Procedure --- p.74 / Chapter 2.9 --- Production of rabbit polyclonal antibodies against isolated stroke protein / Chapter 2.9.1 --- Isolation of stroke protein band from SDS-PAGE slab gel --- p.78 / Chapter 2.9.2 --- Production of anti-stroke protein serum in rabbits --- p.79 / Chapter 2.10 --- Western blotting method / Chapter 2.10.1 --- Reagents --- p.80 / Chapter 2.10.2 --- Procedures --- p.81 / Chapter 2.11 --- Extraction of total cellular RNA by lithium chloride method / Chapter 2.11.1 --- Reagents --- p.83 / Chapter 2.11.2 --- Procedures --- p.84 / Chapter 2.11.3 --- Checking the purity of the extracted RNA --- p.85 / Chapter 2.12 --- Purification of mRNA / Chapter 2.12.1 --- Reagents --- p.85 / Chapter 2.12.2 --- Procedure --- p.86 / Chapter 2.13 --- Verification of purity of mRNA / Chapter 2.13.1 --- Reagents --- p.87 / Chapter 2.13.2 --- Procedure --- p.88 / Chapter 2.14 --- Translation of gerbil brain mRNA in reticulocyte lysates and analysis of its product by SDS PAGE / Chapter 2.14.1 --- Reagents --- p.89 / Chapter 2.14.2 --- Procedures --- p.89 / Chapter CHAPTER THREE --- ESTABLISHMENT OF AN ANIMAL STROKE MODEL / Chapter 3.1 --- Foreword --- p.92 / Chapter 3.2 --- Preliminary studies / Chapter 3.2.1 --- Introduction --- p.92 / Chapter 3.2.2 --- Results --- p.93 / Chapter 3.2.3 --- Discussion --- p.96 / Chapter 3.3 --- Survival rate analysis / Chapter 3.3.1 --- Introduction --- p.97 / Chapter 3.3.2 --- Result --- p.98 / Chapter 3.3.3 --- Discussion --- p.102 / Chapter 3.4 --- Neurologic signs of ischemia / Chapter 3.4.1 --- Introduction --- p.103 / Chapter 3.4.2 --- Result --- p.105 / Chapter 3.4.3 --- Discussion --- p.111 / Chapter 3.5 --- Ocular fundus examination / Chapter 3.5.1 --- Introduction --- p.112 / Chapter 3.5.2 --- Result --- p.114 / Chapter 3.5.3 --- Discussion --- p.116 / Chapter 3.6 --- Inclined plane method / Chapter 3.6.1 --- Introduction --- p.117 / Chapter 3.6.2 --- Result --- p.118 / Chapter 3.6.3 --- Discussion --- p.121 / Chapter 3.7 --- Histologic examination using TTC as staining agent / Chapter 3.7.1 --- Introduction --- p.122 / Chapter 3.7.2 --- Result --- p.124 / Chapter 3.7.3 --- Discussion --- p.129 / Chapter CHAPTER FOUR --- IDENTIFICATION OF ALTERED PROTEIN PATTERN IN THE - BRAINS OF STROKE GERBILS BY ELECTROPHORETIC METHODS / Chapter 4.1 --- Separation of soluble brain extracts by SDS-PAGE analysis / Chapter 4.1.1 --- Introduction --- p.130 / Chapter 4.1.2 --- Result --- p.132 / Chapter 4.1.3 --- Discussion --- p.140 / Chapter 4.2 --- Two dimensional electrophoretic analysis of soluble brain extracts from stroke gerbils / Chapter 4.2.1 --- Introduction --- p.142 / Chapter 4.2.2 --- Result --- p.143 / Chapter 4.2.3 --- Discussion --- p.148 / Chapter CHAPTER FIVE --- ISOLATION OF STROKE-ASSOCIATED PROTEIN FROM BRAINS OF STROKE GERBILS BY IMMUNOCHEMICAL METHOD / Chapter 5.1 --- Introduction --- p.149 / Chapter 5.2 --- Result --- p.151 / Chapter 5.3 --- Discussion --- p.153 / Chapter CHAPTER SIX --- DETECTION OF NEW PROTEIN TRANSLATED FROM MESSENGER RIBONUCLEIC ACID FROM BRAINS OF STROKE GERBIL / Chapter 6.1 --- Introduction / Chapter 6.1.1 --- Extraction of stroke gerbil brain messenger ribonucleic acid --- p.154 / Chapter 6.1.2 --- Translation of mRNA --- p.154 / Chapter 6.2 --- Results / Chapter 6.2.1 --- Yield of total cellular RNA --- p.157 / Chapter 6.2.2 --- Verification of purity of mRNA --- p.157 / Chapter 6.2.3 --- Autoradiographic patterns of translated proteins --- p.159 / Chapter 6.3 --- Discussion --- p.163 / Chapter CHAPTER SEVEN --- GENERAL DISCUSSION --- p.165 / BIBLIOGRAPHY --- p.168

Wong Pak Ki. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 86-88). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.iv / Dedication --- p.v / Table of Contents --- p.vi / List of Abbreviations --- p.ix / List of Appendices --- p.xi / List of Figures --- p.xiv / List of Tables --- p.xx / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1 --- Ginseng and Ginsenosides --- p.1 / Chapter 1.2 --- Instrumental Analysis of Ginsenosides --- p.6 / Chapter 1.2.1 --- Thin Layer Chromatography --- p.6 / Chapter 1.2.2 --- Infrared Spectroscopy --- p.7 / Chapter 1.2.3 --- Colorimetry --- p.7 / Chapter 1.2.4 --- Gas Chromatography --- p.7 / Chapter 1.2.5 --- High Performance Liquid Chromatography --- p.8 / Chapter 1.3 --- Objective of the Study --- p.9 / Chapter Chapter 2: --- Experimental --- p.13 / Chapter 2.1 --- History of Electrophoresis and Capillary Electrophoresis --- p.13 / Chapter 2.1.1 --- Electroosmotic Flow (EOF) --- p.14 / Chapter 2.1.2 --- Electrophoretic Migration --- p.18 / Chapter 2.2 --- Reagents and Materials --- p.20 / Chapter 2.2.1 --- Reagents and Glassware --- p.20 / Chapter 2.2.2 --- Instrumentation --- p.20 / Chapter 2.2.3 --- Preparation of Solutions and Wavelength Selection --- p.22 / Chapter 2.2 --- Procedures --- p.23 / Chapter Chapter 3: --- Results and Discussions --- p.24 / Chapter 3.1 --- Initial Selection of the Running Electrolyte --- p.24 / Chapter 3.2 --- Inclusion Additives in the Aqueous Buffer Solution --- p.29 / Chapter 3.2.1 --- Reasons for Addition of Buffer Additives --- p.29 / Chapter 3.2.1.1 --- Cyclodextrin --- p.29 / Chapter 3.3 --- Addition of Surfactants --- p.33 / Chapter 3.3.1 --- Sodium Dodecyl Sulfate (SDS) --- p.35 / Chapter 3.3.2 --- Sodium Cholate --- p.41 / Chapter 3.4 --- Addition of Organic Modifier --- p.43 / Chapter 3.5 --- Effect of pH --- p.46 / Chapter 3.6 --- Effect of the Concentration of the Borate/Phosphate Solution --- p.51 / Chapter 3.7 --- Effect of Capillaries with Different Inner Diameters (I.D.) --- p.54 / Chapter 3.7.1 --- Effect of pH --- p.54 / Chapter 3.7.2 --- Effect of the Buffer Concentration --- p.60 / Chapter 3.7.3 --- Comparison of Migration Time between Capillaries of 50μm and 75μm Inner Diameter --- p.62 / Chapter 3.8 --- Optimization of Other Experimental Parameters --- p.66 / Chapter 3.8.1 --- Applied Voltage --- p.66 / Chapter 3.8.2 --- The Time of Injection --- p.68 / Chapter 3.8.3 --- The Operating Temperature --- p.70 / Chapter 3.9 --- Intra-day and Inter-day Reproducibility --- p.72 / Chapter 3.10 --- Quantitative Analysis of the Ginsenosides --- p.74 / Chapter 3.11 --- Application of the Developed Methodology --- p.78 / Chapter 3.11.1 --- Experimental Procedures --- p.79 / Chapter Chapter 4: --- Conclusion --- p.83 / References --- p.86 / Appendices --- p.89

by Cheung Yuk Yin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves xx-xxviii). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 摘要 --- p.iii / Table of Content --- p.iv / Abbreviations / for Amino Acids --- p.viii / for Standard Genetic Code --- p.ix / for Units --- p.x / for Prefixes --- p.xi / for Terms commonly used in the report --- p.xii / List of Figures --- p.xiii / List of Tables --- p.xiv / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Epidemiology of Hepatitis B Virus (HBV) --- p.1 / Chapter 1.2 --- Relationship between Hepatitis B Virus and Hepatocellular Carcinoma --- p.2 / Chapter 1.3 --- Brief Description of HBV Genome --- p.2 / Chapter 1.4 --- Possible Roles of HBx in Hepatocellular Carcinoma --- p.4 / Chapter 1.5 --- Novel Interacting Partner of HBx - HBx-lnteracting Protein (XIP) --- p.6 / Chapter 1.6 --- Objective --- p.6 / Chapter Chapter 2 --- Methodology / Chapter 2.1 --- Information of the HBx and XIP Clones --- p.7 / Chapter 2.2 --- "Information of the Expression Vectors (pRSETA, 6xHis-pRSETA and pET8C)" --- p.7 / Chapter 2.3 --- Sub-Cloning of HBx and XIP into Different Vectors --- p.9 / Chapter 2.3.1 --- Design of Primers for Cloning of HBx and XIP into Different Vectors --- p.9 / Chapter 2.3.2 --- Polymerase Chain Reaction (PCR) Protocol --- p.12 / Chapter 2.3.3 --- Enzyme Digestion Reaction Protocol --- p.14 / Chapter 2.3.4 --- Ligation Protocol --- p.16 / Chapter 2.3.5 --- Preparation of Competent Cells --- p.17 / Chapter 2.3.6 --- Transformation --- p.18 / Chapter 2.3.7 --- Gel Extraction Protocol --- p.19 / Chapter 2.3.7.1 --- Life Technologies CONCERT´ёØ Rapid Gel Extraction System --- p.19 / Chapter 2.3.7.2 --- QIAGEN Gel Extraction Kit --- p.20 / Chapter 2.3.8 --- Plasmid Preparation Protocol --- p.22 / Chapter 2.3.8.1 --- Life Technologies CONCERT´ёØ Rapid Plasmid Minipreps --- p.22 / Chapter 2.3.8.2 --- QIAGEN Plasmid Maxi Kit --- p.23 / Chapter 2.4 --- Expression of HBx and XIP in E. coli Strain C41 (DE3) --- p.25 / Chapter 2.4.1 --- Transformation --- p.25 / Chapter 2.4.2 --- Expression of HBx and 6xHis-HBx in E. coli Strain C41 (DE3) --- p.26 / Chapter 2.4.3 --- Expression of XIP in E. coli Strain C41 (DE3) --- p.27 / Chapter 2.5 --- Preparation of Buffers for Chromatography and Circular Dichroism Spectrum Measurement --- p.28 / Chapter 2.6 --- Purification and Refolding of HBx and His-Tagged HBx --- p.28 / Chapter 2.6.1 --- Washing of HBx and His-Tagged HBx Inclusion Bodies --- p.28 / Chapter 2.6.2 --- His-Tagged HBx Purification by Affinity Chromatography --- p.29 / Chapter 2.6.3 --- HBx Purification by Size Exclusion Chromatography --- p.30 / Chapter 2.6.4 --- Refolding of HBx and His-Tagged HBx by Oxidative Dialysis --- p.30 / Chapter 2.7 --- Purification of XIP --- p.33 / Chapter 2.7.1 --- Screening of Chromatographic Conditions for the Purification of XIP --- p.33 / Chapter 2.7.2 --- XIP 1st Step of Purification by Hydrophobic Interaction Chromatography --- p.34 / Chapter 2.7.3 --- XIP 2nd step of Purification by Size Exclusion Chromatography --- p.34 / Chapter 2.8 --- Chemical Denaturation Experiment of HBx and XIP --- p.36 / Chapter 2.8.1 --- Preparation of Urea Buffers for the Chemical Denaturation of HBx --- p.37 / Chapter 2.8.2 --- Preparation of Different GdnHCI Buffer for the Chemical Denaturation of XIP --- p.38 / Chapter 2.8.3 --- Calculation for Chemical Denaturation Experiment --- p.39 / Chapter 2.8.3.1 --- Protein Concentration Calculation --- p.39 / Chapter 2.8.3.2 --- Residual Molar Elipticity Calculation --- p.39 / Chapter 2.8.3.3 --- Free Energy Change (ΔGu) Calculation --- p.40 / Chapter 2.9 --- Two-dimensional Heteronuclear Nuclear Magnetic Resonance (NMR) Experiment --- p.41 / Chapter 2.10 --- Interaction Confirmation between HBx and XIP --- p.42 / Chapter 2.10.1 --- "Transfection of pEGFP, pEGFP-HBx and pEGFP-XIP into HepG2" --- p.42 / Chapter 2.10.2 --- Yeast Two Hybrid System for Confirmation of HBx and XIP Interaction --- p.44 / Chapter 2.10.2.1 --- Preparation of Y187 Competent Cells --- p.44 / Chapter 2.10.2.2 --- Transformation of pGBKT7-HBx and pACT2-XIP into Y187 --- p.45 / Chapter 2.10.2.3 --- β-galactosidase Colony Lift Assay --- p.46 / Chapter Chapter 3 --- "Expression, Purification and Characterization of Hepatitis B Virus X Protein (HBx)" / Chapter 3.1 --- Introduction --- p.47 / Chapter 3.2 --- Construction of Recombinant HBx-pRSETA and 6xHis-HBx-pRSETA Plasmids --- p.48 / Chapter 3.3 --- Expression of 6xHis-HBx in E. coli C41 (DE3) using M9ZB Medium --- p.52 / Chapter 3.4 --- Expression of HBx in E. coli C41 (DE3) using M9ZB Medium --- p.54 / Chapter 3.5 --- Purification and Refolding of 6xHis-HBx Fusion Proteins --- p.56 / Chapter 3.6 --- Purification and Refolding of HBx Proteins --- p.60 / Chapter 3.7 --- Structural Characterization of Refolded HBx --- p.65 / Chapter 3.7.1 --- Introduction --- p.55 / Chapter 3.7.2 --- Experimental Analysis of HBx Secondary Structure --- p.66 / Chapter 3.7.3 --- Chemical Unfolding Experiment of HBx --- p.68 / Chapter 3.8 --- Discussion --- p.70 / Chapter 3.8.1 --- "HBx was Expressed, Purified and Characterized instead of 6xHis-HBx" --- p.71 / Chapter 3.8.2 --- High Concentration of DTT was used to Minimize Formation of HBx Aggregates --- p.72 / Chapter 3.8.3 --- Oxidative Refolding to Ensure Proper Disulfide Bond Formation --- p.73 / Chapter 3.8.4 --- Computational Prediction and Experimental Prediction of Secondary Structure of HBx --- p.75 / Chapter 3.9 --- Concluding Remarks --- p.77 / Chapter Chapter 4 --- "Expression, Purification and Characterization of HBx-lnteracting Protein (XIP)" / Chapter 4.1 --- Introduction --- p.78 / Chapter 4.2 --- Construction of Recombinant XIP-pET8C --- p.78 / Chapter 4.3 --- Expression of XIP in E. coli C41 (DE3) using M9ZB and M9 Mediums --- p.82 / Chapter 4.4 --- Screening of Chromatographic Conditions for the Purification of XIP --- p.83 / Chapter 4.4.1 --- Introduction --- p.83 / Chapter 4.4.2 --- Purification Details --- p.83 / Chapter 4.5 --- Purification of XIP by HiTrap Phenyl HP 5-ml Column --- p.87 / Chapter 4.6 --- Purification of XIP by HiLoad 26/60 Superdex 75 Prep Grade --- p.89 / Chapter 4.7 --- Structural Characterization of XIP --- p.92 / Chapter 4.7.1 --- CD Spectrum --- p.92 / Chapter 4.7.2 --- Chemical Denaturation Experiment of XIP --- p.93 / Chapter 4.7.3 --- Two-Dimensional Heteronuclear Nuclear Magnetic Resonance (NMR) Spectrum of 15N Labeled XIP --- p.95 / Chapter 4.8 --- Discussion --- p.97 / Chapter 4.8.1 --- Purification Method Development --- p.97 / Chapter 4.8.2 --- "Do Different Protein Cosolutes, Protein Stabilizers and Detergents Help XIP to Adopt a Stable Conformation?" --- p.99 / Chapter 4.9 --- Concluding Remarks --- p.101 / Chapter Chapter 5 --- In vivo Studies of HBx and XIP Interactions / Chapter 5.1 --- Investigation of Sub-Cellular Localization of HBx and XIP in Liver Cells --- p.102 / Chapter 5.1.1 --- Introduction --- p.102 / Chapter 5.1.2 --- "Construction of Recombinant HBx-pECFP-C1, HBx-pEGFP-C1, HBx-pEYFP-C1 and XIP-pECFP-C1, XIP-pEGFP-C1, XIP-pEYFP-C1" --- p.103 / Chapter 5.1.3 --- Transfection of pEGFP-C1 HBx and pEGFP-C1 XIP into HepG2 to Find Out HBx and XIP Sub-Cellular Localization --- p.106 / Chapter 5.1.3.1 --- Introduction --- p.107 / Chapter 5.1.3.2 --- Investigation of EGFP Proteins Expression using the Confocal Microscope and the Leica TCS Software --- p.108 / Chapter 5.1.4 --- Discussion and Future Prospects --- p.111 / Chapter 5.2 --- Interaction of HBx and XIP Studied by Yeast Two-Hybrid System --- p.113 / Chapter 5.2.1 --- Introduction --- p.113 / Chapter 5.2.2 --- Construction of Recombinant HBx-pGBKT7 and XIP-pACT2 Plasmids --- p.114 / Chapter 5.2.3 --- Confirmation of HBx and XIP Interaction by Yeast Two-Hybrid System --- p.117 / Chapter 5.2.4 --- Discussion --- p.121 / Chapter Chapter 6 --- Conclusion --- p.123 / Appendix I Sequence of HBx and XIP --- p.I / Chapter II --- Vector Sequences --- p.II / Chapter III --- Vector Maps --- p.VI / Chapter IV --- Electrophoresis Markers --- p.XI / Chapter V --- Agarose Gel Electrophoresis --- p.XII / Chapter VI --- SDS-PAGE Eectrophoresis --- p.XIII / Chapter VII --- Medium for Bacterial Culture --- p.XV / Chapter VIII --- Medium for Cell Culture --- p.XVII / Chapter IX --- Medium for Yeast Culture --- p.XVIII / Chapter X --- Buffers for Yeast Transformation --- p.XIX / Reference --- p.XX

Ip Yee-man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 85-89). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Abstract --- p.iii / Table of Contents --- p.vi / Abbreviations --- p.xi / List of Figures --- p.xiii / List of Tables --- p.xvii / Chapter / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Background --- p.1 / Chapter 1.2 --- Quality control of Chinese herbal medicine --- p.2 / Chapter 1.3 --- Rhizoma Chuanxiong --- p.4 / Chapter 1.3.1. --- General description --- p.4 / Chapter 1.3.2. --- Chemical constituents --- p.4 / Chapter 1.3.3. --- Pharmacology --- p.7 / Chapter 1.3.4 --- Instrumental analysis --- p.9 / Chapter 1.3.4.1 --- Thin Layer Chromatography (TLC) --- p.9 / Chapter 1.3.4.2 --- Gas Chromatography (GC) --- p.10 / Chapter 1.3.4.3 --- High Performance Liquid Chromatography (HPLC) --- p.10 / Chapter 1.3.4.4 --- Capillary Electrophoresis (CE) --- p.10 / Chapter 1.4 --- Objectives of the study --- p.11 / Chapter 2. --- "Isolation, Characterization and Identification of Reference Compounds" --- p.13 / Chapter 2.1 --- General experiment procedures --- p.13 / Chapter 2.1.1. --- Solvents for chromatographic separation --- p.13 / Chapter 2.1.2 --- Chromatographic methods --- p.13 / Chapter 2.1.2.1 --- Adsorption column chromatography --- p.13 / Chapter 2.1.2.2 --- Thin layer chromatography --- p.13 / Chapter 2.1.2.3 --- Preparative layer chromatography --- p.14 / Chapter 2.1.3 --- Determination of physical data --- p.14 / Chapter 2.1.3.1 --- Infrared (IR) absorption spectra --- p.14 / Chapter 2.1.3.2 --- Nuclear Magnetic Resonance (NMR) spectra --- p.14 / Chapter 2.1.3.3 --- Mass spectra (MS) --- p.15 / Chapter 2.1.3.4 --- X-ray crystallography --- p.15 / Chapter 2.1.4 --- Authentic reference compounds --- p.15 / Chapter 2.2 --- "Procurement, extraction and initial fractionation of Rhizoma Chuanxiong" --- p.15 / Chapter 2.3 --- Chromatographic separation of the chloroform extract --- p.16 / Chapter 2.3.1 --- Chromatographic separation of fraction F1002 --- p.16 / Chapter 2.3.1.1 --- Spectral data for the characterization of compound 1 [5-(hydroxymethyl)- 2- furancarboxaldehyde] --- p.17 / Chapter 2.3.2 --- Column chromatographic separation of fraction F1003A --- p.17 / Chapter 2.3.2.1 --- Spectral data for the characterization of compound 2 (oleic acid) --- p.18 / Chapter 2.3.2.2 --- Physical data for the characterization of compound 3 (ferulic acid) --- p.18 / Chapter 2.3.3 --- Preparative layer chromatographic separation of Fraction F1010 --- p.19 / Chapter 2.3.3.1 --- Spectral data for the characterization of compound 4 (daucosterol) --- p.19 / Chapter 2.4 --- Column chromatographic separation of the hexane extract --- p.19 / Chapter 2.4.1 --- Removal of fatty acids in fraction F2005 and F2006 by partition --- p.20 / Chapter 2.4.2 --- Column chromatographic separation of fraction F2005M --- p.20 / Chapter 2.4.2.1 --- Spectral data for the characterization of compound 5 (butylidenephthalide) --- p.20 / Chapter 2.4.2.2 --- Spectral data for the characterization of compound 6 (butylphthalide) --- p.21 / Chapter 2.4.3 --- Column chromatographic separation of fraction F2006M --- p.21 / Chapter 2.4.3.1 --- "Spectral data for the characterization of compound 7 (Z, Z'-6.6', 7.3'a- diligustilide)" --- p.21 / Chapter 2.4.4 --- Colum chromatographic separation of fraction --- p.22 / Chapter 2.4.4.1 --- Spectral data for the characterization of compound 8 (pregnenolone) --- p.22 / Chapter 2.4.4.2 --- "Spectral data for the characterization of compound 9 [5,5- oxydimethylenebis(2-furaldehyde)]" --- p.23 / Chapter 2.5 --- Results and Discussion --- p.24 / Chapter 2.5.1 --- Identification of compound 1 [5-(hydroxymethyl)-2- furancarboxaldehyde] --- p.24 / Chapter 2.5.2 --- Identification of compound 2 (oleic acid) --- p.25 / Chapter 2.5.3 --- Identification of compound 3 (ferulic acid) --- p.26 / Chapter 2.5.4 --- Identification of compound 4 (daucosterol) --- p.26 / Chapter 2.5.5 --- Identification of compound 5 (butylidenephthalide) --- p.27 / Chapter 2.5.6 --- Identification of compound 6 (butylphthalide) --- p.28 / Chapter 2.5.7 --- "Identification of compound 7 (Z, Z'-6.6', 7.3'a-diligustilide)" --- p.30 / Chapter 2.5.8 --- Identification of compound 8 (pregnenolone) --- p.31 / Chapter 2.5.9 --- "Identification of compound 9 [5,5'-oxydimethylenebis(2-furaldehyde)]" --- p.32 / Chapter 2.6 --- Conclusions --- p.34 / Chapter 3. --- Analysis of Rhizoma Chuanxiong by Capillary Electrophoresis --- p.35 / Chapter 3.1 --- Introduction --- p.35 / Chapter 3.1.1 --- Capillary electrophoreis system --- p.35 / Chapter 3.1.2 --- Principles of separation --- p.36 / Chapter 3.1.3 --- Considerations on development of analysis method --- p.41 / Chapter 3.2 --- Experimental --- p.43 / Chapter 3.2.1 --- Reagents and materials --- p.43 / Chapter 3.2.2 --- Reference compounds --- p.43 / Chapter 3.2.3 --- Instrumentation and apparatus --- p.44 / Chapter 3.2.4 --- Experimental procedures --- p.45 / Chapter 3.2.4.1 --- Preparation of running buffer solution --- p.45 / Chapter 3.2.4.2 --- Preparation of standard solutions --- p.46 / Chapter 3.2.4.3 --- Preparation of Rhizoma Chuanxiong extracts --- p.47 / Chapter 3.2.4.4 --- Flushing of capillaries --- p.47 / Chapter 3.2.4.5 --- Conditions of separation --- p.48 / Chapter 3.3 --- Results and Discussion --- p.48 / Chapter 3.3.1 --- Preliminary experiments --- p.48 / Chapter 3.3.1.1 --- Addition of surfactants --- p.51 / Chapter 3.3.2 --- Effect of buffer concentration --- p.54 / Chapter 3.3.3 --- Effect of SDS concentration --- p.59 / Chapter 3.3.4 --- Addition of organic modifier --- p.63 / Chapter 3.3.5 --- Reproducibility of the proposed method --- p.68 / Chapter 3.3.6 --- Quantitative analysis of seven standard compounds --- p.70 / Chapter 3.3.7 --- Application of the developed methodology --- p.74 / Chapter 3.3.8 --- Conclusions --- p.83 / References --- p.85 / Appendices / Appendix 1.1.1 1H-NMR spectrum of 5-(hydroxymethyl)-2-furancarboxaldehyde --- p.90 / Appendix 1.1.2 13C-NMR spectrum of 5-(hydroxyinethyl)-2-furancarboxaldehyde --- p.90 / Appendix 1.2 X-ray crystallographic data of ferulic acid --- p.91 / Appendix 1.3 13C-NMR spectrum of butylidenephthalide --- p.96 / Appendix 1.4.1 1 H-NMR spectrum of butylphthalide --- p.97 / Appendix 1.4.2 13C-NMR spectrum of butylphthalide --- p.97 / "Appendix 1.5 X-ray crystallographic data of z, z', 6.6'， 7.3'a-diligustilide" --- p.98 / "Appendix 1.6 X-ray crystallographic data of 5,5'-oxydimethylenebis(2-furaldehyde)" --- p.105 / Appendix 2.1 Details of quantitative analysis of 5-(hydroxymethyl)-2-furancarboxaldehyde --- p.112 / Appendix 2.2 Details of quantitative analysis of ligustrazin hydrochloride --- p.112 / "Appendix 2.3 Details of quantitative analysis of 5,5'-oxydimethylenebis(2-furaldehyde)" --- p.113 / Appendix 2.4 Details of quantitative anlaysis of ferulic acid --- p.113 / Appendix 2.5 Details of quantitative analysis of butylphthalide --- p.114 / Appendix 2.6 Details of quantitative analysis of butylidenephthalide --- p.114 / "Appendix 2.7 Details of quantitative anlaysis of z,z', 6.6', 7.3'a-diligustilide" --- p.115 / Appendix 3.1 Quantitative analysis of Chuanxiong sample from Hong Kong (HK1) --- p.115 / Appendix 3.2 Quantitaive analysis of Chuanxiong sample from Hong Kong (HK2) --- p.116 / Appendix 3.3 Quantitative analysis of Chuanxiong sample from Sichuan (SC1) --- p.116 / Appendix 3.4 Quantitative analysis of Chuanxiong sample from Sichuan (SC2) --- p.117 / Appendix 3.5 Quantitative anlaysis of Chuanxiong samplefrom Fujian (FJ) --- p.117

by Ricardo Wong Chi Ho. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 128-138). / Abstracts in English and Chinese. / Acknowledgments --- p.i / Abstract --- p.ii / 論文摘要 --- p.iv / Table of Contents --- p.vi / List of Figures --- p.xi / List of Tables --- p.xiii / List of Abbreviations --- p.xiv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Overview of Serine Protease Inhibitors --- p.1 / Chapter 1.2 --- Classification of Serine Protease Inhibitors --- p.2 / Chapter 1.2.1 --- Kunitz Type Serine Protease Inhibitors --- p.7 / Chapter 1.2.2 --- Bowman-Birk Type Serine Protease Inhibitors --- p.11 / Chapter 1.2.3 --- Squash Type Serine Protease Inhibitors --- p.16 / Chapter 1.3 --- Role of Serine Protease Inhibitors in Plants --- p.20 / Chapter 1.4 --- Nutritional Fact of Serine Protease Inhibitors --- p.22 / Chapter 1.5 --- Possible Applications of Serine Protease Inhibitors --- p.25 / Chapter 1.5.1 --- Medical Applications --- p.25 / Chapter 1.5.2 --- Agricultural Applications --- p.29 / Chapter 1.6 --- Rationale of the Present Study --- p.31 / Chapter Chapter 2 --- Screening of Seeds for Inhibitory Activities Against Serine Proteases --- p.33 / Chapter 2.1 --- Introduction --- p.33 / Chapter 2.2 --- Materials and Methods --- p.37 / Chapter 2.2.1 --- Materials --- p.37 / Chapter 2.2.2 --- Extraction Method --- p.37 / Chapter 2.2.3 --- Assays for Proteases Inhibitory Activities --- p.38 / Chapter 2.2.3.1 --- Assay for Chymotrypsin Activity --- p.38 / Chapter 2.2.3.2 --- Assay for Trypsin Activity --- p.38 / Chapter 2.2.3.3 --- Assay for Elastase Activity --- p.39 / Chapter 2.2.3.4 --- Assay for Subtilisin Activity --- p.39 / Chapter 2.2.3.5 --- Assays for Protease Inhibitory Activities --- p.40 / Chapter 2.2.4 --- Determination of Protein Concentration --- p.41 / Chapter 2.3 --- Results --- p.42 / Chapter 2.3.1 --- Extraction --- p.42 / Chapter 2.3.2 --- Serine Proteases Inhibitory Activities --- p.42 / Chapter 2.4 --- Discussion --- p.47 / Chapter Chapter 3 --- Isolation of Chymotrypsin Inhibitor and Trypsin Inhibitors from Momordica cochinchinensis Seeds --- p.49 / Chapter 3.1 --- Introduction --- p.49 / Chapter 3.2 --- Materials and Methods --- p.56 / Chapter 3.2.1 --- Materials --- p.56 / Chapter 3.2.2 --- Protein Extraction --- p.57 / Chapter 3.2.3 --- SP-Sepharose Chromatography --- p.57 / Chapter 3.2.4 --- Reversed Phase High Pressure Liquid Chromatography --- p.58 / Chapter 3.2.5 --- Assays for Chymotrypsin and Trypsin Inhibitory Activities --- p.60 / Chapter 3.2.6 --- Titration of Chymotrypsin --- p.61 / Chapter 3.2.7 --- Tricine Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis --- p.62 / Chapter 3.2.8 --- Coupling of Trypsin-Sepharose 4B Affinity Column --- p.63 / Chapter 3.2.9 --- Affinity Chromatography on Trypsin-Sepharose 4B --- p.64 / Chapter 3.3 --- Results --- p.65 / Chapter 3.3.1 --- SP-Sepharose Chromatography --- p.65 / Chapter 3.3.2 --- Reversed Phase High Pressure Liquid Chromatography --- p.67 / Chapter 3.3.3 --- Summary of Purification --- p.71 / Chapter 3.3.4 --- Titration of Chymotrypsin --- p.74 / Chapter 3.3.5 --- Tricine Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis --- p.74 / Chapter 3.3.6 --- Affinity Chromatography on Trypsin-Sepharose 4B --- p.78 / Chapter 3.4 --- Discussion --- p.81 / Chapter Chapter 4 --- Characterization of Chymotrypsin Inhibitor and Trypsin Inhibitors --- p.88 / Chapter 4.1 --- Introduction --- p.88 / Chapter 4.2 --- Materials and Methods --- p.90 / Chapter 4.2.1 --- Materials --- p.90 / Chapter 4.2.2 --- Determination of Molecular Weight --- p.90 / Chapter 4.2.3 --- Amino Acid Sequence Analysis --- p.91 / Chapter 4.2.4 --- Surface Plasmon Resonance Measurement --- p.92 / Chapter 4.2.4.1 --- Immobilization of Ligands on the Surface of Optical Biosensors --- p.92 / Chapter 4.2.4.2 --- Determination of Kinetics Constants --- p.93 / Chapter 4.2.4.3 --- pH Dependence of the Inhibition by Chymotrypsin Inhibitor --- p.93 / Chapter 4.2.4.4 --- Data Analysis --- p.94 / Chapter 4.2.5 --- Effect of Chymotrypsin Inhibitor on the Estereolytic Activity and Proteolytic Activity of Chymotrypsin --- p.95 / Chapter 4.2.6 --- Specificities of the Inhibitors % --- p.96 / Chapter 4.2.7 --- Binding Ratio of CI to Different Proteases --- p.97 / Chapter 4.2.8 --- Effects of the Proteases on Their Corresponding Inhibitors --- p.97 / Chapter 4.2.8.1 --- Tricine Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis --- p.97 / Chapter 4.2.8.2 --- Assay for Chymotrypsin Inhibitory Activity --- p.98 / Chapter 4.3 --- Results --- p.99 / Chapter 4.3.1 --- Molecular Weight of the Inhibitors --- p.99 / Chapter 4.3.2 --- N-terminal Amino Acid Sequence --- p.99 / Chapter 4.3.3 --- Surface Plasmon Resonance Measurement --- p.102 / Chapter 4.3.3.1 --- Kinetics of Chymotrypsin Inhibitor --- p.102 / Chapter 4.3.3.2 --- Kinetics of Trypsin Inhibitors --- p.106 / Chapter 4.3.3.3 --- pH Dependence of the Inhibition by Chymotrypsin Inhibitor --- p.106 / Chapter 4.3.4 --- Effect of Chymotrypsin Inhibitor on the Estereolytic Activity and Proteolytic Activity of Chymotrypsin --- p.106 / Chapter 4.3.5 --- Specificities of the Inhibitors --- p.110 / Chapter 4.3.6 --- Binding Ratio of CI to Different Proteases --- p.112 / Chapter 4.3.7 --- Effects of the Proteases on Their Corresponding Inhibitors --- p.112 / Chapter 4.4 --- Discussion --- p.119 / Chapter Chapter 5 --- Conclusion --- p.125 / References --- p.128

by Choy Ka-Fai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 107-125). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.I / 論文摘要 --- p.II / ABSTRACT --- p.III / ABBREVIATIONS --- p.V / TABLE OF CONTENTS --- p.VI / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter CHAPTER 2 --- PURIFICATION OF GRASS CARP ALDH FROM MITOCHONDRIA --- p.18 / Chapter CHAPTER 3 --- PURIFICATION & CHARACTERIZATION OF GRASS CARP ALDH --- p.49 / Chapter CHAPTER 4 --- CONCLUSION --- p.104 / REFERENCES --- p.107

by Tang Wai Kwan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 91-110). / Abstracts in English and Chinese. / Acknowledgements / 論文摘要 / Abstract / Abbreviations / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Aldehyde Dehydrogenase Extended Family --- p.1 / Chapter 1.1.1 --- Phylogenetic Tree --- p.2 / Chapter 1.1.2 --- Physiological Functions --- p.4 / Chapter 1.1.3 --- Structural Conservations --- p.7 / Chapter 1.2 --- ALDH-1 and ALDH-2 --- p.9 / Chapter 1.3 --- Antiquitin --- p.11 / Chapter 1.4 --- Osmoregulation --- p.14 / Chapter 1.4.1 --- Osmoprotectant --- p.14 / Chapter 1.4.2 --- Betaine Aldehyde Dehydrogenase --- p.15 / Chapter 1.5 --- Objectives of the Present Study --- p.18 / Chapter Chapter 2 --- Purification and Characterization of Seabream ALDH-2 and Antiquitin --- p.20 / Chapter 2.1 --- Introduction --- p.20 / Chapter 2.2 --- Materials --- p.21 / Chapter 2.3 --- Methodology / Chapter 2.3.1 --- Preparation of Crude Tissue Extract --- p.22 / Chapter 2.3.2 --- Synthesis of α-Cyanocinnamate Sepharose --- p.22 / Chapter 2.3.3 --- Synthesis of p-Hydroxyacetophenone Sepharose --- p.23 / Chapter 2.3.4 --- Purification of ALDH-2 --- p.23 / Chapter 2.3.5 --- Purification of Antiquitin --- p.24 / Chapter 2.3.6 --- Enzyme and Protein Assays --- p.24 / Chapter 2.3.7 --- Electrophoretic Procedures / Chapter 2.3.7.1 --- Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.26 / Chapter 2.3.7.2 --- Native PAGE --- p.27 / Chapter 2.3.7.3 --- Isoelectric focusing (IEF) --- p.27 / Chapter 2.3.8 --- N-terminal Amino Acid Sequencing --- p.28 / Chapter 2.4 --- Results / Chapter 2.4.1 --- Tissue Distribution of ALDH --- p.29 / Chapter 2.4.2 --- Purification and Molecular Properties of ALDH-2 --- p.31 / Chapter 2.4.3 --- Kinetic Properties of ALDH-2 --- p.42 / Chapter 2.4.4 --- Purification and Molecular Properties of Antiquitin --- p.49 / Chapter 2.4.5 --- Kinetic Properties of Antiquitin --- p.54 / Chapter Chapter 3 --- Discussion / Chapter 3.1 --- Tissue Distribution --- p.66 / Chapter 3.2 --- N-terminal Amino Acid Sequencing --- p.67 / Chapter 3.3 --- Purification of Seabream ALDH --- p.68 / Chapter 3.3.1 --- Separation of Two ALDH isoforms --- p.69 / Chapter 3.3.2 --- Binding Affinity of α-Cyanocinnamate Sepharose --- p.70 / Chapter 3.3.3 --- Purification --- p.72 / Chapter 3.4 --- Electrophoretic Properties --- p.73 / Chapter 3.5 --- pH and Temperature Stability --- p.74 / Chapter 3.6 --- Substrate Specificity --- p.77 / Chapter 3.7 --- Possible Functions of Antiquitin --- p.80 / Chapter 3.8 --- Future Prospects --- p.84 / Chapter Chapter 4 --- Conclusion --- p.90 / Chapter Chapter 5 --- References --- p.91

Chen, Yu. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 75-78). / Abstracts in English and Chinese. / Isolation and Identification of Differentially Expressed Protein in Serum of Patients with Sleep Disorders --- p.I / Abstract --- p.IV / 論文摘要 --- p.VII / Acknowledgements --- p.IX / Table of Contents --- p.X / List of Figures --- p.XII / List of Tables --- p.XII / List of Abbreviations --- p.XIII / Chapter Chapter 1: --- Introduction --- p.2 / Chapter 1.1 --- Definition of narcolepsy --- p.2 / Chapter 1.2 --- Symptoms of narcolepsy --- p.2 / Chapter 1.2.1 --- Excessive Daytime Sleepiness (EDS) --- p.2 / Chapter 1.2.2 --- Cataplexy --- p.2 / Chapter 1.2.3 --- Associated features --- p.3 / Chapter 1.3 --- Prevalence of narcolepsy --- p.4 / Chapter 1.4 --- Pathophysiology and molecular genetics of narcolepsy --- p.7 / Chapter 1.4.1 --- Pathophysiology of narcolepsy --- p.7 / Chapter 1.4.2 --- Molecular genetics research --- p.8 / Chapter 1.5 --- Diagnostic criteria for narcolepsy --- p.12 / Chapter 1.6 --- Treatment of narcolepsy --- p.16 / Chapter 1.7 --- The Burden of narcolepsy --- p.18 / Chapter 1.8 --- Human blood serum/plasma --- p.19 / Chapter 1.9 --- Cerebrospinal fluid (CSF) --- p.23 / Chapter 1.10 --- Aims of study --- p.26 / Chapter Chapter 2: --- Materials and Methods --- p.28 / Chapter 2.1 --- Participants and measurements --- p.28 / Chapter 2.1.1 --- Participants --- p.28 / Chapter 2.1.2 --- Diagnosis measurements --- p.28 / Chapter 2.2 --- "Serum extraction, albumin and IgG depletion" --- p.30 / Chapter 2.2.1 --- Albumin and IgG Depletion Kit --- p.30 / Chapter 2.2.2 --- Chemicals and reagents --- p.30 / Chapter 2.2.3 --- Preparation of solutions --- p.30 / Chapter 2.2.4 --- Procedure --- p.30 / Chapter 2.3 --- Reversed Phase High Performance Liquid Chromatography (RP-HPLC) --- p.32 / Chapter 2.3.1 --- RP-HPLC method --- p.32 / Chapter 2.3.2 --- Chemicals and reagents --- p.33 / Chapter 2.3.3 --- Preparation of mobile phases --- p.33 / Chapter 2.3.4 --- Procedure --- p.33 / Chapter 2.4 --- MALDI-TOF/TOF Mass Spectrometry --- p.35 / Chapter 2.4.1 --- Chemicals and reagents --- p.35 / Chapter 2.4.2 --- Preparation of solutions --- p.35 / Chapter 2.4.3 --- Procedure --- p.35 / Chapter 2.5 --- SDS-PAGE and double staining --- p.37 / Chapter 2.5.1 --- Chemicals and reagents --- p.37 / Chapter 2.5.2 --- Preparation of solutions --- p.37 / Chapter 2.5.3 --- Procedure --- p.39 / Chapter 2.6 --- N-terminal amino acid analysis --- p.42 / Chapter 2.6.1 --- Procedure --- p.42 / Chapter 2.6.2 --- Sequence analysis --- p.42 / Chapter 2.7 --- CSF analysis --- p.43 / Chapter Chapter 3: --- Results --- p.45 / Chapter 3.1 --- Albumin and IgG depletion of human serum samples --- p.45 / Chapter 3.2 --- Peak identification --- p.47 / Chapter 3.2.1 --- Peak identification on HPLC profiles --- p.47 / Chapter 3.2.2 --- Statistical results --- p.51 / Chapter 3.2.3 --- Family cases analysis --- p.54 / Chapter 3.3 --- MALDI-TOF/TOF Mass Spectrometry --- p.56 / Chapter 3.4 --- SDS-PAGE and double staining --- p.58 / Chapter 3.5 --- Protein sequence analysis --- p.60 / Chapter 3.6 --- Cerebrospinal fluid (CSF) analysis --- p.62 / Chapter Chapter 4: --- Discussion --- p.65 / Chapter 4.1 --- RP-HPLC methods --- p.65 / Chapter 4.2 --- The detected peptide fragment and Hlark --- p.66 / Chapter 4.2.1 --- "Human Lark protein (Hlark, hlark)" --- p.66 / Chapter 4.2.2 --- Circadian clocks --- p.67 / Chapter 4.2.3 --- "Hlark, circadian rhythm and narcolepsy" --- p.71 / Chapter 4.3 --- Familial and genetic analysis --- p.72 / Chapter 4.4 --- Clinical implications --- p.73 / Chapter 4.5 --- Conclusion --- p.74 / References --- p.75

Yau Wing Yiu, Winifred. / On t.p. "alpha"s appear as the Greek letter. / Thesis submitted in: October 2006. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 91-98). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract (Chinese version) --- p.iv / Acknowledgements --- p.vi / Table of Contents --- p.vii / List of Abbreviations --- p.xii / List of Figures --- p.xiv / List of Tables --- p.xvi / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Peroxisome proliferator-activated receptors (PPARs) --- p.1 / Chapter 1.1.1 --- What are PPARs? --- p.1 / Chapter 1.1.2 --- PPAR ligands - peroxisome proliferators --- p.1 / Chapter 1.1.3 --- PPAR isoforms --- p.2 / Chapter 1.2 --- Biological roles of PPARα --- p.3 / Chapter 1.2.1 --- Lipid metabolism --- p.3 / Chapter 1.2.2 --- Glucose metabolism --- p.4 / Chapter 1.2.3 --- Inflammation --- p.5 / Chapter 1.2.4 --- Oxidative stress --- p.5 / Chapter 1.2.5 --- Cell proliferation and apoptosis --- p.6 / Chapter 1.3 --- PPARα in health and diseases --- p.6 / Chapter 1.3.1 --- Wound-healing --- p.6 / Chapter 1.3.2 --- Anti-atherogenesis --- p.7 / Chapter 1.3.3 --- Neuroprotection --- p.7 / Chapter 1.3.4 --- Carcineogenesis --- p.7 / Chapter 1.4 --- PPARα-regulated and starvation inducible gene (PPSIG) --- p.8 / Chapter 1.4.1 --- PPSIG is a PPARα target gene --- p.8 / Chapter 1.4.2 --- Computer-assisted predictions on PPSIG --- p.9 / Chapter 1.4.3 --- Current characterization of PPSIG --- p.10 / Chapter 1.5 --- Objectives of the present study --- p.11 / Chapter Chapter 2 --- Materials and Methods --- p.12 / Chapter 2.1 --- Materials --- p.12 / Chapter 2.2 --- Animals and treatment --- p.13 / Chapter 2.3 --- Cloning of PPSIG into pThioHis and pTYB expression vectors --- p.13 / Chapter 2.3.1 --- PCR amplification of PPSIG cDNA insert --- p.13 / Chapter 2.3.1.1 --- PPSIG cDNA insert for pThioHis vector --- p.13 / Chapter 2.3.1.2 --- PPSIG cDNA insert for pTYB vector --- p.15 / Chapter 2.3.2 --- Restriction enzyme digestion of PPSIG cDNA insert and pThioHis vector --- p.18 / Chapter 2.3.3 --- Restriction enzyme digestion of PPSIG cDNA insert and pTYB vector --- p.20 / Chapter 2.3.4 --- Ligation and transformation --- p.20 / Chapter 2.3.5 --- Screening for recombinants by phenol/chloroform method --- p.21 / Chapter 2.3.6 --- Confirmation of recombinant plasmid by restriction enzyme digestion --- p.22 / Chapter 2.3.6.1 --- Digestion of pThioHis-PPSIG plasmid with Xba I and Sac II --- p.22 / Chapter 2.3.6.2 --- Digestion of pTYB-PPSIG plasmid with EcoR V --- p.22 / Chapter 2.3.7 --- Transformation into expression E. coli strains --- p.23 / Chapter 2.4 --- Over expression of PPSIG proteins in E. coli --- p.23 / Chapter 2.5 --- Semi-purification of PPSIG fusion proteins by preparative SDS-PAGE --- p.24 / Chapter 2.6 --- Rabbit immunization --- p.25 / Chapter 2.7 --- Northern blotting analysis --- p.26 / Chapter 2.7.1 --- Probe preparation --- p.26 / Chapter 2.7.2 --- "Formaldehyde-agarose gel electrophoresis, blotting of RNA and hybridization" --- p.26 / Chapter 2.8 --- Subcellular fractionation --- p.29 / Chapter 2.9 --- Western blotting of liver microsomes --- p.31 / Chapter 2.10 --- Immunoprecipitation --- p.32 / Chapter 2.11 --- Mass spectrometry --- p.33 / Chapter 2.11.1 --- Trypsin digestion and peptide extraction --- p.33 / Chapter 2.11.2 --- Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry --- p.34 / Chapter Chapter 3 --- Results --- p.36 / Chapter 3.1 --- Cloning of PPSIG into pThioHis and pTYB vectors --- p.36 / Chapter 3.1.1 --- Cloning of PPSIG into pThioHis vector --- p.36 / Chapter 3.1.2 --- Cloning of PPSIG into pTYB vector --- p.36 / Chapter 3.2 --- Protein expression of Thio-PPSIG and Intein-PPSIG --- p.41 / Chapter 3.3 --- Identification of recombinant Thio-PPSIG and Intein-PPSIG by mass spectrometry --- p.49 / Chapter 3.4 --- Preparation and characterization of Thio-PPSIG and Intein-PPSIG antisera --- p.61 / Chapter 3.5 --- Identification of native PPSIG and its induction pattern --- p.65 / Chapter 3.5.1 --- PPSIG was highly inducible upon 72-h starvation in a PPARα dependent manner --- p.65 / Chapter 3.5.2 --- "PPSIG showed slight induction upon 2-wk Wy-14,643 treatment" --- p.71 / Chapter 3.6 --- Confirmation of the specificity of PPSIG antiserum --- p.74 / Chapter Chapter 4 --- Discussion --- p.81 / References --- p.91 / Appendix A Deduced amino acid sequences of PPSIG fusion proteins --- p.99 / Chapter A1 --- Deduced amino acid sequence of Thio-PPSIG from pThioHis-PPSIG plasmid --- p.99 / Chapter A2 --- Deduced amino acid sequence of Intein-PPSIG from pTYB-PPSIG plasmid --- p.101 / Appendix B Mass spectra of trypsin digested native PPSIG --- p.104 / Chapter B1 --- Mass spectrum of trypsin digested native PPSIG immunoprecipitated from liver microsomes from PPARα wild-type mice fed with normal diet (starvation experiment) --- p.104 / Chapter B2 --- Mass spectrum of trypsin digested native PPSIG immunoprecipitated from liver microsomes from PPARα wild-type mice starved for 72 hours (starvation experiment) --- p.105 / Chapter B3 --- "Mass spectrum of trypsin digested native PPSIG immunoprecipitated from liver microsomes from PPARα wild-type mice fed with control diet (Wy-14,643 feeding experiment)" --- p.106 / Chapter B4 --- "Mass spectrum of trypsin digested native PPSIG immunoprecipitated from liver microsomes from PPARα wild-type mice fed with 0.1% (w/w) Wy-14,643 for 2 weeks (Wy-14,643 feeding experiment)" --- p.107

Stroke är den vanligast förekommande sjukdomen i Sverige med cirka 30 000 insjuknanden per år. Ur ett svenskt och internationellt perspektiv är stroke den största orsaken till en icke medfödd funktionsnedsättning hos vuxna individer. Motorisk funktionsnedsättning efter stroke påverkar patienters förmåga att utföra aktiviteter i dagliga livet (ADL). Det inverkar på patienters upplevelse av livet och livskvalité. En begränsning av kroppskontroll medför även en förändrad relation till kroppen. År 2012 var cirka 20 procent av patienter beroende av stöd i ADL, tre månader efter insjuknandet i stroke. Studiens syfte är att beskriva upplevelser av påverkan på ADL hos strokepatienter med nedsatt motorisk funktion. Studien är en litteraturöversikt och bygger på kvalitetsgranskade kvalitativa primärkällor. Databaserna Cinahl och PubMed har använts för att identifiera vetenskapliga artiklar publicerade tidigast år 2000. Det resulterade i att nio kvalitativa artiklar med västerländskt ursprung inkluderades till studien. Fem teman framträdde ur artiklarna vilka utgör grunden för resultatet: upplevelse av att plötsligt drabbas av stroke, upplevelse av att vara isolerad, upplevelse av att vara beroende, upplevelse av otrygghet och upplevelse av förändrad identitet. Tidigare forskning bekräftar studiens resultat och beskriver insjuknandet i stroke via upplevelser av utsatthet och av att inte vara samma person. Forskningen lyfter även vikten att vara i ett betydelsefullt sammanhang. De negativa upplevelserna som patienter beskriver kan tänkas begränsas genom insatser från samhället och ett värdigt bemötande från sjuksköterskor med patienters livsvärld i fokus. / Program: Sjuksköterskeutbildning