Wong, Ngan Yuk. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 114-128). / Abstracts in English and Chinese. / Thesis/Assessment Committee --- p.i / Declaration --- p.ii / Abstract (in English) --- p.iii / Abstract (in Chinese) --- p.v / Acknowledgments --- p.vi / Table of Contents --- p.vii / List of Abbreviations --- p.xi / List of Figures --- p.xiv / List of Tables --- p.xvi / Chapter 1. --- Literature review --- p.1 / Chapter 1.1. --- Introduction to Cordyceps --- p.1 / Chapter 1.2. --- Ingredients of Cordyceps and their related biological activities --- p.4 / Chapter 1.2.1. --- "Amino acids, peptides, proteins and polyamines" --- p.4 / Chapter 1.2.1.1. --- Proteins --- p.4 / Chapter 1.2.2. --- Saccharides and sugar derivatives --- p.7 / Chapter 1.2.2.1. --- Polysaccharides --- p.7 / Chapter 1.2.3. --- Nucleosides --- p.9 / Chapter 1.2.3.1. --- Cordycepin --- p.9 / Chapter 1.2.3.2. --- Adenosine --- p.12 / Chapter 1.2.4. --- Fatty acids and sterols --- p.14 / Chapter 1.2.5. --- Vitamins and inorganics --- p.15 / Chapter 1.3. --- Cordyceps and their related biological activities --- p.15 / Chapter 1.3.1. --- Cordyceps militaris --- p.15 / Chapter 1.3.2. --- Cordyceps sinensis --- p.17 / Chapter 1.4. --- Proteomic tools used to study the change in protein expression profiles --- p.21 / Chapter 1.4.1. --- Proteomic tools in studies of the change in protein expression --- p.21 / Chapter 1.4.2. --- Two-dimensional gel electrophoresis --- p.22 / Chapter 1.4.3. --- Mass spectrometry --- p.22 / Chapter 1.4.4. --- Current challenges --- p.23 / Chapter 2. --- Methodology --- p.25 / Chapter 2.1. --- Cultivation of Cordyceps militaris --- p.25 / Chapter 2.2. --- Preparation of Cordyceps extracts for anti-proliferation assay on cell lines --- p.26 / Chapter 2.2.1. --- Types of the extracts of Cordyceps --- p.26 / Chapter 2.2.2. --- Preparation of the Cordyceps extracts --- p.26 / Chapter 2.3. --- Anti-proliferation assay on cell lines for extract screening --- p.26 / Chapter 2.3.1. --- Cell lines and culturing condition --- p.26 / Chapter 2.3.2. --- Viable cell count using trypan blue exclusion method --- p.27 / Chapter 2.3.3. --- Anti-proliferation assay on cell lines using MTT assay --- p.28 / Chapter 2.3.4. --- Determination of the IC50 values --- p.30 / Chapter 2.3.5. --- Statistical Analysis --- p.30 / Chapter 2.4. --- Proteomic studies for HepG2 and Hs68 after the treatment of extracts --- p.30 / Chapter 2.4.1. --- Protein sample preparation of HepG2and Hs68 --- p.30 / Chapter 2.4.2. --- Protein quantitation --- p.31 / Chapter 2.4.3. --- 2D Gel electrophoresis --- p.33 / Chapter 2.4.4. --- Image analysis --- p.34 / Chapter 2.4.5. --- In gel digestion and MALDI-ToF MS --- p.35 / Chapter 2.5. --- Cell cycle analysis --- p.36 / Chapter 2.5.1. --- Cell samples preparation --- p.36 / Chapter 2.5.2. --- Popidium iodide staining --- p.36 / Chapter 2.5.3. --- Flow cytometry --- p.37 / Chapter 2.5.4. --- Statistical Analysis --- p.38 / Chapter 2.6. --- Western blotting --- p.38 / Chapter 2.6.1. --- Protein sample preparation of HepG2 and Hs68 --- p.38 / Chapter 2.6.2. --- SDS-PAGE --- p.38 / Chapter 2.6.3. --- Protein Transblotting --- p.39 / Chapter 2.6.4. --- Membrane Blocking and Antibody Incubations --- p.39 / Chapter 2.6.5. --- Detection of Proteins --- p.40 / Chapter 3. --- Results --- p.41 / Chapter 3.1. --- Investigation of anti-proliferating effect of Cordyceps extracts on HepG2 and Hs68 using MTT assays --- p.41 / Chapter 3.1.1. --- Cordyceps militaris fruiting body extract - CMFB --- p.41 / Chapter 3.1.2. --- Cordyceps militaris mycelia extract - CMM --- p.41 / Chapter 3.1.3. --- Cordyceps sinensis fruiting body extract - CSFB --- p.45 / Chapter 3.1.4. --- Cordyceps sinensis mycelia extract - CSM --- p.45 / Chapter 3.1.5. --- "Comparison of the anti-proliferation effects of the Cordyceps extracts CMFB, CMM, CSFB and CSM" --- p.49 / Chapter 3.2. --- "Investigation of anti-proliferating effect of Cordyceps militaris extracts on H292, Neuro2a and WIL2-NS using MTT assays" --- p.51 / Chapter 3.2.1. --- Cordyceps militaris fruiting body extract - CMFB --- p.51 / Chapter 3.2.2. --- Cordyceps militaris mycelia extract - CMM --- p.51 / Chapter 3.3. --- Changes in total protein expression profiles in cell lines --- p.56 / Chapter 3.3.1. --- Protein samples preparation --- p.56 / Chapter 3.3.2. --- 2D gel electrophoresis analysis of protein from cell lines --- p.56 / Chapter 3.3.2.1. --- HepG2 --- p.57 / Chapter 3.3.2.2. --- Hs68 --- p.58 / Chapter 3.3.3. --- Differentially expressed proteins identification --- p.66 / Chapter 3.3.3.1. --- HepG2 --- p.66 / Chapter 3.3.3.2. --- Hs68 --- p.67 / Chapter 3.4. --- Cell cycle analysis --- p.76 / Chapter 3.4.1. --- Cell samples preparation --- p.76 / Chapter 3.4.2. --- HepG2 --- p.76 / Chapter 3.4.3. --- Hs68 --- p.77 / Chapter 3.4.4. --- H292 --- p.77 / Chapter 3.5. --- Western blotting --- p.81 / Chapter 3.5.1. --- Protein samples preparation --- p.81 / Chapter 3.5.2. --- Detection of actin for protein loading normalization --- p.83 / Chapter 3.5.3. --- Detection of procaspase-3 and cleaved caspase-3 --- p.83 / Chapter 3.5.4. --- Detection of procaspase-7 and cleaved caspase-7 --- p.84 / Chapter 3.5.5. --- Detection of procaspase-9 and cleaved caspase-9 --- p.84 / Chapter 4. --- Discussion --- p.86 / Chapter 4.1. --- Anti-tumor proliferating effect of Cordyceps extracts --- p.86 / Chapter 4.2. --- Changes in total protein expression profiles in cell lines --- p.87 / Chapter 4.2.1. --- Differentially expressed proteins in HepG2 treated with fruiting body extract --- p.88 / Chapter 4.2.1.1. --- Heat shock 90kDa protein 1 beta (HSP90(β) --- p.89 / Chapter 4.2.1.2. --- Far upstream element-binding protein 1 (FUBP-1) --- p.90 / Chapter 4.2.1.3. --- RuvB-like 1 (RuvbLl) --- p.90 / Chapter 4.2.1.4. --- Acidic protein rich in leucine (APRIL) --- p.91 / Chapter 4.2.1.5. --- SET protein --- p.92 / Chapter 4.2.1.6. --- Enolase 1 (α-enolase) --- p.94 / Chapter 4.2.1.7. --- Aldolase A --- p.96 / Chapter 4.2.1.8. --- DNA-binding protein B --- p.96 / Chapter 4.2.1.9. --- Peroxiredoxin 1 (Prx 1) --- p.97 / Chapter 4.2.1.10. --- Proteasome activator subunit 1 iso form 1 --- p.99 / Chapter 4.2.1.11. --- Dehydrogenase/ reductase member 2 isoform 2 --- p.99 / Chapter 4.2.1.12. --- Protein disulfide isomerase- related protein 5 --- p.100 / Chapter 4.2.1.13. --- Annexin IV --- p.100 / Chapter 4.2.1.14. --- Enoyl Coenzyme A hydratase --- p.101 / Chapter 4.2.2. --- Differentially expressed proteins in HepG2 treated with mycelia extract --- p.102 / Chapter 4.2.2.1. --- Alpha actinin 4 --- p.102 / Chapter 4.2.2.2. --- SET translocation isoform 1 --- p.103 / Chapter 4.2.2.3. --- Acidic (leucine-rich) nuclear phosphoprotein 32 family member B (ANP32b) --- p.103 / Chapter 4.2.2.4. --- Endoplasmic reticulum protein 29 isoform 1 precursor (ERp29) --- p.103 / Chapter 4.2.2.5. --- Heterogeneous nuclear ribonucleoprotein H3 isoform b (hnRNP 2H9A) --- p.104 / Chapter 4.2.3. --- Differentially expressed proteins in Hs68 treated with fruiting body extract --- p.105 / Chapter 4.2.3.1. --- Lamin A/C isoform 2 --- p.105 / Chapter 4.2.3.2. --- Vimentin --- p.106 / Chapter 4.2.3.3. --- Tropomyosin 1 alpha chain isoform 4 --- p.107 / Chapter 4.2.3.4. --- Rho GDP dissociation inhibitor (GDI) alpha (RhoGDIα) --- p.109 / Chapter 4.2.3.5. --- Dihydropyrimidinase-like 2 (DRP-2) --- p.109 / Chapter 4.2.3.6. --- Keratin 7 (K7) --- p.110 / Chapter 4.2.4. --- Differentially expressed proteins in Hs68 treated with mycelia extract --- p.111 / Chapter 4.3. --- Cell cycle analysis --- p.111 / Chapter 4.4. --- Western blotting --- p.113 / Chapter 5. --- References --- p.114
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326996 |
| Date | January 2010 |
| Contributors | Wong, Ngan Yuk., Chinese University of Hong Kong Graduate School. Division of Life Sciences. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xvi, 128 leaves : ill. ; 30 cm. |
| Rights | Use 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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