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

This reseach firstly investigated iodine-induced apoptotic effects and the underlying mechanism in thyroid cancer cells. Results indicated that apoptosis induced by iodine, especially at high dose of iodine (100 muM), was mitochondrial-mediated, with the loss of mitochondrial membrane potential, Bak up-regulation, caspase 3 activation and cytochrome C release from mitochondria. Iodine treatment decreased the level of mutant p53 including the R273H mutant that possesses anti-apoptotic features while increased the p21 level. The block of p21 significantly prevented iodine-induced apoptosis. High doses of iodine also stimulated the transient activation of the subfamily members of MAPKs (ERK1/2, p38 and JNK1/2). The results showed the three subfamily members of MAPKs all worked as anti-apoptotic factors. Surprisingly, high doses of iodine promoted instead of suppressed the expression of anti-apoptotic protein Bcl-xL expression. The increase of Bc1-xL was likely to compensate the damage induced by iodine since the inhibition of Bc1-xL accelerated iodine-mediated apoptosis. Collectively, iodine induced mitochondrial-mediated apoptosis in thyroid cancer cells. This apoptotic pathway was involved in the activation of MAPKs pathways, which may subsequently up-regulate p21, Bc1-xL, and down-regulate anti-apoptotic mutant p53 expression. The findings provide solid molecular evidence to explain the epidemiological observation that iodine insufficiency promotes the thyroid tumor development. It may also reveal some novel molecular targets for the treatment of thyroid cancer. / Thyroid cancer is the most common endocrine malignancy and exhibits the full range of malignant behaviors from the relatively indolent occult differentiated thyroid cancer to uniformly aggressive and lethal anaplastic thyroid cancer. Iodine is a well known key element in thyroid normal function maintenance and thyroid cancer development. However, the mechanisms of iodine in thyroid cancer cells development are limited. Recent researches have indicated that iodine could induce cancer cells apoptosis, staying clear from the dysfunction of iodide-specific transportation systems in thyroid cancer cells. Thus, iodine-induced apoptosis may be an effective pathway for iodine to affect thyroid cancer development, but we know little about them. / To further explore iodine on the apoptotic effects of chemotherapeutic agents in thyroid cancer, anaplastic thyroid cancer cell line ARO was used. Anaplastic thyroid cancer is lethal because of its rapid progression and poor response to chemotherapy and radioiodine therapy. The study examined the effect of moderate dose of iodine (50 muM) on the apoptosis of ARO cells treated with doxorubicin (Dox) and histone deacetylase inhibitor sodium butyrate (NaB). The cytotoxic effect of either Dox or NaB alone was limited, but co-administration of NaB and Dox (NaB-Dox) significantly increased mitochondrial-mediated apoptosis. The effects of iodine to apoptosis-induced by the two agents were diversified. Iodine reduced the apoptosis induced by Dox or NaB-Dox but promoted apoptosis induced by NaB. To explain this diversifying finding, the experiment found that iodine exaggerated NaB-mediated Bcl-xL down-regulation. In contrast, it reduced the effect of Dox on the decrease of Bcl-xL expression. Further experiments showed that iodine regulated the level of Bcl-xL in ERK- or/and p38-related pathways. The balance between ERK and p38 may determine the iodine-modulated Bcl-xL expression. The high ERK/p38 activity ratio up-regulated Bc1-xL and enabled the tumor cells to resist chemotherapy, whereas the low ERK/p38 down-regulated Bc1-xL and sensitized the tumor cells to chemotherapy. Taken together, iodine plays a critical role in apoptosis of thyroid cancer cells induced by chemotherapeutic agents. The balance between ERK and p38 may determine cell survival and death through modulating Bcl-xL expression in thyroid cancer cells. The findings provide some new insights into the roles of iodine in chemotherapeutic agents-induced apoptosis in thyroid cancer cells. / To summarize, iodine-induced apoptotic effects on thyroid cancer cells is a key pathway for iodine to influence thyroid cancer development and chemotherapy. Meanwhile MAPKs-related mutant p53, p21 and Bcl-xL expression are critical in deciding thyroid cancer cells survival and death. Moreover, iodine can influence chemotherapeutic agents-induced apoptosis through ERK/p38-mediated Bcl-xL expression. / Liu, Xiaohong. / "December 2009." / Adviser: Charles Andrew van Hasselt. / Source: Dissertation Abstracts International, Volume: 72-01, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 111-146). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Leung, Ching Hei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 101-110). / Abstracts in English and Chinese. / Acknowledgements --- p.i / 摘要（Abstract in Chinese) --- p.iii / Abstract --- p.v / List of Abbreviations --- p.vii / List of Figures and Tables --- p.ix / Table of Content --- p.xi / Chapter CHAPTER 1 --- Introduction / Chapter 1.1 --- History and development of photodynamic therapy --- p.1 / Chapter 1.2 --- Basic principle of photodynamic therapy: the beauty of the treatment --- p.3 / Chapter 1.3 --- "Photosensitizers: From discovery, synthesis to modifications" --- p.6 / Chapter 1.4 --- Enhancement of selective retention of PS in cancerous tissue --- p.10 / Chapter 1.5 --- Development of silicon (IV) phthalocyanine derivatives --- p.14 / Chapter 1.6 --- Death mechanisms in photodynamic therapy --- p.17 / Chapter 1.7 --- Objectives of the present study --- p.18 / Chapter CHAPTER 2 --- Materials and Methods / Chapter 2.1 --- Synthesis of BAM-SiPc --- p.20 / Chapter 2.2 --- Preparation of BAM-SiPc solution for photodynamic treatment --- p.20 / Chapter 2.3 --- Cell line and culture conditions --- p.21 / Chapter 2.4 --- Animal tumor model --- p.23 / Chapter 2.5 --- PDT laser source --- p.23 / Chapter 2.6 --- In vitro photodynamic activity assay --- p.23 / Chapter 2.6.1 --- Preparation of cells for photodynamic treatment / Chapter 2.6.2 --- In vitro photodynamic treatment / Chapter 2.6.3 --- Cell viability assay / Chapter 2.7 --- "Determination of reactive oxygen species production by 2',7'- dichlorofluorescein diacetate (DCFDA) assay" --- p.28 / Chapter 2.8 --- Analysis of cell cycle arrest --- p.28 / Chapter 2.9 --- Biodistribution of BAM-SiPc --- p.29 / Chapter 2.10 --- In vivo photodynamic treatment --- p.30 / Chapter 2.11 --- Assay for plasma enzyme activities --- p.30 / Chapter 2.12 --- Determination of cellular uptake of BAM-SiPc --- p.31 / Chapter 2.13 --- Metabolism of BAM-SiPc --- p.31 / Chapter 2.14 --- Histochemical staining --- p.32 / Chapter 2.14.1 --- Preparation of paraffin-embedded tissue section / Chapter 2.14.2 --- Haematoxylin and Eosin (H & E) staining / Chapter 2.14.3 --- Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay / Chapter 2.15 --- Conjugation of BAM-SiPc with LDL --- p.34 / Chapter 2.15.1 --- Analysis of the phototoxicity and cellular uptake of BAM- SiPc in the presence of LDL / Chapter 2.15.2 --- Gel filtration analysis of the mixture of LDL and BAM- SiPc / Chapter 2.16 --- Statistical analysis --- p.35 / Chapter CHAPTER 3 --- Results / Chapter 3.1 --- In vitro photodynamic activity assays --- p.36 / Chapter 3.2 --- Tissue distribution of BAM-SiPc in HepG2- bearing nude mice --- p.39 / Chapter 3.3 --- Anti-tumor activities of in vivo PDT with BAM-SiPc --- p.42 / Chapter 3.3.1 --- In vivo effect of PDT treatment with BAM-SiPc on HepG2 and HT29 tumor growth / Chapter 3.3.2 --- Dosage effect on anti-tumor activities by BAM-SiPc mediated PDT / Chapter 3.4 --- Analysis of intrinsic toxicity induced by BAM-SiPc mediated PDT --- p.48 / Chapter 3.4.1 --- H & E staining of liver sections of nude mice after in vivo PDT / Chapter 3.4.2 --- Plasma enzyme activity assays of PDT treated mice / Chapter 3.5 --- BAM-SiPc metabolism in in vitro culture cells and liver homogenate --- p.53 / Chapter 3.5.1 --- Cellular uptake of BAM-SiPc / Chapter 3.5.2 --- BAM-SiPc metabolism in cultured normal liver cells and cancer cells / Chapter 3.5.3 --- BAM-SiPc metabolism by mice liver homogenate / Chapter 3.6 --- Death mechanism induced by BAM-SiPc mediated PDT --- p.62 / Chapter 3.6.1 --- Events related to cell death induced by in vitro BAM-SiPc mediated PDT / Chapter 3.6.2 --- Death mechanism exerted by in vivo BAM-SiPc mediated PDT / Chapter 3.7 --- Effect on phototoxicity of BAM-SiPc in the presence of LDL --- p.70 / Chapter 3.7.1 --- Effect on phototoxicity of BAM-SiPc after mixing BAM- SiPc with LDL / Chapter 3.7.2 --- Gel filtration for analysis of the LDL-BAM-SiPc mixture / Chapter CHAPTER 4 --- Discussion / Chapter 4.1 --- Anti-cancer effect of BAM-SiPc on different cancer cell lines --- p.76 / Chapter 4.2 --- Tissue distribution of BAM-SiPc in HepG2 bearing nude mice --- p.77 / Chapter 4.3 --- In vivo effect of BAM-SiPc mediated PDT on HepG2 and HT29 tumor growth --- p.80 / Chapter 4.4 --- Analysis of the safety of using BAM-SiPc as a potential agent in PDT --- p.83 / Chapter 4.5 --- Metabolism of BAM-SiPc --- p.84 / Chapter 4.6 --- Mechanism of the apoptosis triggered by BAM-SiPc mediated PDT --- p.88 / Chapter 4.7 --- Death mechanism induced by in vivo PDT with BAM-SiPc --- p.93 / Chapter 4.8 --- Phototoxicity of BAM-SiPc in the presence of LDL --- p.94 / Chapter CHAPTER 5 --- Conclusion and Future perspective / Chapter 5.1 --- Conclusion --- p.97 / Chapter 5.2 --- Future perspective --- p.98 / References

Lai, Sze Tsai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 94-104). / Abstracts in English and Chinese. / Thesis Committees --- p.i / Statement --- p.ii / Abstract --- p.iii / 摘要 --- p.v / Acknowledgments --- p.vi / List of Abbreviations --- p.vii / Table of Contents --- p.ix / List of Tables --- p.xiii / List of Figures --- p.xiv / Chapter 1 --- Literature review --- p.1 / Chapter 1.1 --- Introduction to Cordyceps --- p.1 / Chapter 1.2 --- Fungal proteomics --- p.2 / Chapter 1.2.1 --- Extraction method --- p.2 / Chapter 1.2.2 --- Proteomic study of Cordyceps --- p.3 / Chapter 1.3 --- Ingredients of Cordyceps and their related biological activities --- p.5 / Chapter 1.3.1 --- Polysaccharides --- p.5 / Chapter 1.3.2 --- Nucleosides --- p.6 / Chapter 1.3.2.1 --- Cordycepin --- p.6 / Chapter 1.3.2.2 --- Adenosine --- p.8 / Chapter 1.4 --- Cordyceps and their related biological activities --- p.9 / Chapter 1.4.1 --- Cordyceps militaris --- p.9 / Chapter 1.4.2 --- Cordyceps sinensis --- p.10 / Chapter 1.5 --- Proteomic analysis of proteome change --- p.12 / Chapter 1.5.1 --- Proteomic tools used to study the change in protein expression --- p.12 / Chapter 1.5.2 --- Two-dimensional gel electrophoresis --- p.13 / Chapter 1.5.3 --- Mass spectrometry --- p.13 / Chapter 1.6 --- Objective --- p.16 / Chapter 2 --- Methodology --- p.17 / Chapter 2.1 --- Cultivation of Cordyceps militaris --- p.17 / Chapter 2.2 --- Proteomic study on Cordyceps militaris --- p.17 / Chapter 2.2.1 --- Extraction of proteins from Cordyceps militaris --- p.17 / Chapter 2.2.2 --- Protein quantification --- p.18 / Chapter 2.2.3 --- 2D Gel electrophoresis --- p.19 / Chapter 2.2.4 --- Image analysis --- p.20 / Chapter 2.2.5 --- In gel digestion and MALDI-ToF MS --- p.20 / Chapter 2.3 --- Preparation of Cordyceps extracts for anti-proliferation assay on cell lines --- p.21 / Chapter 2.3.1 --- Types of the extracts of Cordyceps --- p.21 / Chapter 2.3.2 --- Preparation of the extracts of Cordyceps --- p.21 / Chapter 2.4 --- Anti-proliferation assay on cell lines for extract screening --- p.22 / Chapter 2.4.1 --- Cell lines and culturing condition --- p.22 / Chapter 2.4.2 --- Viable cell count using trypan blue exclusion method --- p.22 / Chapter 2.4.3 --- Anti-proliferation assay on SV7 tert using MTT assay --- p.23 / Chapter 2.4.4 --- Determination of the IC5o values --- p.24 / Chapter 2.4.5 --- Statistical Analysis --- p.24 / Chapter 2.5 --- Anti-proliferation assay on other cell lines using the two screened extracts --- p.24 / Chapter 2.5.1 --- Cell lines and culturing condition --- p.24 / Chapter 2.5.2 --- "Anti-proliferation assay on on HepG2, H292, Neuro2a，WIL2-NS cells using MTT assay" --- p.25 / Chapter 2.6 --- Proteomic studies for SV7tert and Hs68 after the treatment of extracts --- p.25 / Chapter 2.6.1 --- Protein sample preparation of SV7tert and Hs68 --- p.25 / Chapter 2.6.2 --- Protein quantification --- p.26 / Chapter 2.6.3 --- 2D Gel electrophoresis --- p.26 / Chapter 2.6.4 --- Image analysis --- p.26 / Chapter 2.7 --- Western Immunoblotting --- p.26 / Chapter 2.7.1 --- Protein sample preparation of SV7tert and Hs68 --- p.26 / Chapter 2.7.2 --- SDS-PAGE --- p.27 / Chapter 2.7.3 --- Protein Blotting --- p.27 / Chapter 2.7.4 --- Membrane Blocking and Antibody Incubations --- p.28 / Chapter 2.7.5 --- Detection of Proteins --- p.28 / Chapter 3 --- Results --- p.29 / Chapter 3.1 --- Proteins identification in Cordyceps militaris --- p.29 / Chapter 3.1.1 --- 2D gel electrophoresis analysis and resolution --- p.29 / Chapter 3.1.2 --- Identification and categorization of proteins of mycelia and fruiting body of Cordyceps militaris --- p.30 / Chapter 3.2 --- Investigation of anti-proliferating activity of extracts using MTT assays on SV7tert and Hs68 cell lines --- p.44 / Chapter 3.2.1 --- Mycelia extract from Cordyceps militaris --- p.44 / Chapter 3.2.2 --- Fruiting body extract from Cordyceps militaris --- p.44 / Chapter 3.2.3 --- Mycelia extract from Cordyceps sinensis --- p.47 / Chapter 3.2.4 --- Fruiting body extract from Cordyceps sinensis --- p.47 / Chapter 3.2.5 --- Screening of extracts --- p.50 / Chapter 3.3 --- "Investigation of anti-proliferating activity of extracts using MTT assays on HepG2，H292, Neuro2a and WIL2-NS cell lines" --- p.51 / Chapter 3.3.1 --- Mycelia extract from Cordyceps militaris --- p.51 / Chapter 3.3.2 --- Fruiting body extract from Cordyceps militaris --- p.51 / Chapter 3.4 --- Changes in total protein expression profiles in SV7tert and Hs68 cell lines --- p.56 / Chapter 3.4.1 --- Corresponding extract treatment of cell lines --- p.56 / Chapter 3.4.2 --- 2D gel electrophoresis analysis of protein from cells (SV7tert or Hs68) --- p.56 / Chapter 3.4.2.1 --- SV7tert study --- p.57 / Chapter 3.4.2.2 --- Hs68 study --- p.57 / Chapter 3.4.3 --- Protein identification --- p.65 / Chapter 3.4.3.1 --- Changes in protein expressions in SV7tert after mycelia extract treatment --- p.65 / Chapter 3.4.3.2 --- Changes in protein expressions in Hs68 after mycelia extract treatment --- p.65 / Chapter 3.4.3.3 --- Changes in protein expressions in SV7tert after fruiting body extract treatment --- p.66 / Chapter 3.4.3.4 --- Changes in protein expressions in Hs68 after fruiting body extract treatment --- p.66 / Chapter 3.5 --- Western immunoblotting --- p.71 / Chapter 3.5.1 --- Corresponding extract treatment of cell lines --- p.71 / Chapter 3.5.2 --- Normalization of protein loading using anti-actin antibody --- p.73 / Chapter 3.5.3 --- Detection of caspase 3 by use of antibody --- p.74 / Chapter 3.5.4 --- Detection of cleaved caspase 3 by use of antibody --- p.74 / Chapter 4 --- Discussion --- p.77 / Chapter 4.1 --- Identification of proteins in Cordyceps militaris --- p.77 / Chapter 4.2 --- Difficulties in identifying the proteins in Cordyceps militaris --- p.80 / Chapter 4.3 --- Investigation of anti-proliferating activity of extracts --- p.80 / Chapter 4.4 --- Changes in cell total protein expression profiles in SV7tert and Hs68 cell lines --- p.81 / Chapter 4.4.1 --- Protein alterations in SV7tert treated with mycelia extract --- p.82 / Chapter 4.4.1.1 --- Heat shock protein 27 (Hsp27) --- p.82 / Chapter 4.4.1.2 --- Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) --- p.83 / Chapter 4.4.2 --- Protein alterations in Hs68 with mycelia extract treatment --- p.84 / Chapter 4.4.2.1 --- Chain B of triosephosphate isomerase - Triosephophate isomerase 1 --- p.84 / Chapter 4.4.2.2 --- Glutathione transferase --- p.85 / Chapter 4.4.3 --- Protein alterations in SV7tert with fruiting body extract treatment --- p.86 / Chapter 4.4.3.1 --- Calreticulin precusor --- p.86 / Chapter 4.4.3.2 --- Nucleophosmin 1 isoform 2 (B23) --- p.87 / Chapter 4.4.3.3 --- Heat shock 70kDa protein 8 isoform 1 - Heat shock 70kDa protein (Hsp70) --- p.88 / Chapter 4.4.3.4 --- Voltage-dependent anion channel 2 (VDAC2) --- p.89 / Chapter 4.4.3.5 --- "Tumor protein, translationally controlled (TCTP)" --- p.90 / Chapter 4.4.3.6 --- RAN binding protein 1 (RANBP1) --- p.91 / Chapter 4.4.4 --- Protein alteration in Hs68 with mycelia extract treatment --- p.92 / Chapter 4.5 --- Conclusion --- p.93 / References --- p.94

Lin, Ka Yin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 108-122). / Abstracts in English and Chinese. / Abstract (In English) --- p.ii / Abstract (In Chinese) --- p.iii / Acknowledgment --- p.V / Abstracts of Publications --- p.vi / Abbreviations --- p.viii / List of Figures --- p.ix / List of Table --- p.X / Contents --- p.xi / Chapter Chapter I --- Introduction --- p.Page / Chapter 1.1 --- Pluripotent Stem Cell --- p.1 / Chapter 1.1.1 --- Characteristic of pluripotent stem cells --- p.1 / Chapter 1.1.2 --- Origin of pluripotent stem cells --- p.1 / Chapter 1.1.2.1 --- Embryonic carcinoma cells --- p.2 / Chapter 1.1.2.2 --- Embryonic stem cells --- p.2 / Chapter 1.1.2.3 --- Epiblast stem cells --- p.2 / Chapter 1.1.2.4 --- Embryonic germ cells and adult germline stem cells --- p.3 / Chapter 1.1.2.5 --- Induced pluripotent stem cells --- p.3 / Chapter 1.1.3 --- Pluripotency in Embryonic Stem Cells --- p.4 / Chapter 1.1.3.1 --- Extrinsic signal governing pluripotency --- p.5 / Chapter 1.1.3.1.1 --- LIF signaling --- p.5 / Chapter 1.1.3.1.2 --- BMP signaling --- p.6 / Chapter 1.1.3.1.3 --- Other signaling pathways --- p.6 / Chapter 1.1.3.2 --- Intrinsic sternness factors --- p.7 / Chapter 1.1.3.2.1 --- Oct4 Expression in Embryonic Stem cells --- p.7 / Chapter 1.1.3.2.2 --- Sox-2 Expression in Embryonic Stem Cells --- p.8 / Chapter 1.1.3.2.3 --- Nanog Expression in Embryonic Stem Cells --- p.9 / Chapter 1.1.3.2.4 --- "Transcriptional Regulation of Oct-4, Nanog and Sox-2 in Embryonic Stem Cells" --- p.10 / Chapter 1.1.3.2.5 --- Others pluripotent genes --- p.11 / Chapter 1.1.3.2.5.1 --- Utfl --- p.11 / Chapter 1.1.3.2.5.2 --- Rexl --- p.11 / Chapter 1.1.3.2.5.3 --- Esrrb --- p.12 / Chapter 1.1.3.2.5.4 --- Eras --- p.12 / Chapter 1.1.3.2.5.5 --- Tell --- p.12 / Chapter 1.1.3.2.5.6 --- Dnm3tl --- p.13 / Chapter 1.1.3.2.5.7 --- Dppa3 --- p.13 / Chapter 1.1.3.2.5.8 --- Dppa4 --- p.14 / Chapter 1.1.3.2.5.9 --- Dppa5 --- p.14 / Chapter 1.1.3.2.5.10 --- Klf2 --- p.15 / Chapter 1.2 --- Somatic cell reprogramming --- p.16 / Chapter 1.2.1 --- Definition of reprogramming --- p.16 / Chapter 1.2.2 --- The history of reprogramming --- p.16 / Chapter 1.2.2.1 --- Reprogramming by nuclear transfer --- p.17 / Chapter 1.2.2.2 --- Reprogramming by fusion with ES or EC cells --- p.18 / Chapter 1.2.2.3 --- Reprogramming with defined factor --- p.19 / Chapter 1.3 --- Induced pluripotent stem cells --- p.20 / Chapter 1.3.1 --- Transcription factor used for reprogramming to iPS cells --- p.20 / Chapter 1.3.1.1 --- Klf4 --- p.20 / Chapter 1.3.1.2 --- c-Myc --- p.21 / Chapter 1.3.2 --- Cornerstone of iPSC generation --- p.22 / Chapter 1.3.3 --- Major events in the reprogramming process --- p.23 / Chapter 1.3.4 --- Gene delivery systems for ips cell generation --- p.26 / Chapter 1.3.5 --- Culture system for embryonic stem cells and iPSC --- p.28 / Chapter 1.3.4.1 --- Feeder and serum used cell culture system --- p.28 / Chapter 1.3.4.2 --- Serum-free culture condition --- p.29 / Chapter 1.3.5 --- Differentiation potential of iPSC --- p.30 / Chapter 1.3.5.1 --- In vitro stringency tests --- p.30 / Chapter 1.3.5.2 --- In vivo stringency test --- p.30 / Chapter 1.3.5.3 --- In utero stringency test --- p.31 / Chapter 1.4 --- Mouse Lewis lung carcinoma-D 122 --- p.32 / Chapter 1.5 --- Dendritic cell vaccine in cancer immunotherapy --- p.33 / Chapter 1.5 --- Green Fluorescence protein Reporters --- p.35 / Chapter 1.5.1 --- GFP reporters in embryos and stem cell --- p.35 / Chapter 1.5.2 --- copGFP --- p.35 / Chapter 1.6 --- Aim of study --- p.36 / Chapter Chapter II --- Methods and Materials / Chapter 2.1 --- Materials --- p.37 / Chapter 2.1.1 --- Synthetic oligos used in polymerase chain reaction (PCR) --- p.37 / Chapter 2.1.2 --- DNA clones used in the study --- p.39 / Chapter 2.1.3 --- Materials for DNA manipulation --- p.39 / Chapter 2.1.4 --- Materials for RNA manipulation --- p.39 / Chapter 2.1.5 --- Antibodies --- p.40 / Chapter 2.1.6 --- Kits --- p.41 / Chapter 2.1.7 --- Bacteria strain and culture reagents 41 / Chapter 2.1.8 --- Culture media and reagents --- p.42 / Chapter 2.1.8.1 --- General culture media and reagents --- p.42 / Chapter 2.1.8.2 --- Traditional ES medium --- p.42 / Chapter 2.1.8.3 --- Feeder-free Serum-free ESGRO medium --- p.42 / Chapter 2.1.9 --- Cell lines used --- p.43 / Chapter 2.1.10 --- Instrumentation --- p.43 / Chapter 2.2 --- Methods --- p.44 / Chapter 2.2.1 --- Cell culture --- p.44 / Chapter 2.2.1.1 --- Routine cell culture --- p.44 / Chapter 2.2.1.2 --- Resuscitation and culture from frozen stock --- p.44 / Chapter 2.2.1.3 --- Passage of cells --- p.44 / Chapter 2.2.1.4 --- Cryopreservation of cells --- p.45 / Chapter 2.2.1.5 --- Mouse ES cells culture --- p.45 / Chapter 2.2.1.5.1 --- Passage and maintenance of SNL --- p.45 / Chapter 2.2.1.5.2 --- Inactivation and plating of SNLs (Feeder preparation) --- p.45 / Chapter 2.2.1.5.3 --- Cryopreservation (freezing) of SNLs --- p.46 / Chapter 2.2.1.6 --- Mouse ES cells culture in feeder-free culture conditions --- p.46 / Chapter 2.2.1.6.1 --- Preparation of gelatin coated plates --- p.46 / Chapter 2.2.1.6.2 --- Thawing mouse ES cells --- p.46 / Chapter 2.2.1.6.3 --- Passage of mouse ES cells --- p.47 / Chapter 2.2.1.6.4 --- Freezing mouse ES cells --- p.47 / Chapter 2.2.1.7 --- ES cells differentiation-Formation of embryoid bodies (EBs) --- p.47 / Chapter 2.2.1.8 --- Direct differentiation by retinoic acid --- p.48 / Chapter 2.2.1.9 --- Generation of iPS --- p.48 / Chapter 2.2.2 --- Cell transfections --- p.48 / Chapter 2.2.2.1 --- Lipofectamine 2000 transfection --- p.48 / Chapter 2.2.2.2 --- Nucleofection --- p.49 / Chapter 2.2.2.2.1 --- Optimization of nucleofection --- p.49 / Chapter 2.2.2.2.2 --- Nucleofection condition --- p.49 / Chapter 2.2.3 --- Nucleic acid --- p.49 / Chapter 2.2.3.1 --- Genomic DNA isolation --- p.49 / Chapter 2.2.3.2 --- Restriction Enzyme Digestion --- p.50 / Chapter 2.2.3.3 --- RNA and genomic DNA quantification --- p.50 / Chapter 2.2.3.4 --- Reversed transcription polymerase chain reaction (RT-PCR) --- p.50 / Chapter 2.2.3.4.1 --- RNA isolation and Reverse transcription (RT) --- p.50 / Chapter 2.2.3.4.2 --- Polymerase chain reaction (PCR) --- p.51 / Chapter 2.2.3.4.3 --- Real-time polymerase chain reaction (qRT- PCR) --- p.52 / Chapter 2.2.3.5 --- Agarose gel electrophoresis --- p.53 / Chapter 2.2.3.6 --- Genomic PCR for bisulfite sequencing --- p.53 / Chapter 2.2.4 --- Bacteria and Plasmid preparation --- p.54 / Chapter 2.2.4.1 --- Preparation of competent cells --- p.54 / Chapter 2.2.4.2 --- Heat-shock transformation --- p.54 / Chapter 2.2.4.3 --- Midi prep of plasmid --- p.54 / Chapter 2.2.5 --- Cell Staining --- p.55 / Chapter 2.2.5.1 --- Alkaline phosphatase staining --- p.55 / Chapter 2.2.5.2 --- Immunofluorescence --- p.55 / Chapter 2.2.6 --- Flow cytometry --- p.56 / Chapter 2.2.7 --- Animal Handling --- p.56 / Chapter Chapter III --- Results / Chapter 3.1 --- Generation of Nanog-reporter-GFP-D 122 --- p.57 / Chapter 3.2 --- Nucleofection optimization for D122 --- p.60 / Chapter 3.3 --- Generation ofD122-iPS --- p.65 / Chapter 3.3.1 --- Plasmid construct used in the study --- p.65 / Chapter 3.3.2 --- Protocol of D122-iPS generation --- p.67 / Chapter 3.3.3 --- Reprogramming Efficiency of D12´2ؤreNanog cells --- p.69 / Chapter 3.4 --- Expression of pluripotency markers upon reprogramming --- p.70 / Chapter 3.4.1 --- Alkaline Phosphatase staining --- p.70 / Chapter 3.4.2 --- Nanog-GFP expression --- p.72 / Chapter 3.4.3 --- Pluripotency gene expression upon reprogramming --- p.74 / Chapter 3.4.4 --- GFP positive D122 reNanog Colonies --- p.79 / Chapter 3.5 --- Characterization of the D122-iPS-lC --- p.80 / Chapter 3.5.1 --- Morphology of D122-iPS-lC --- p.80 / Chapter 3.5.2 --- Pluripotency gene expression --- p.82 / Chapter 3.5.3 --- Pluripotency markers SSEA-1 and Oct4 --- p.85 / Chapter 3.5.4 --- Bisulfite genomic sequencing --- p.88 / Chapter 3.5.5 --- Differentiation of the D122-iPS-lC --- p.90 / Chapter 3.5.5.1 --- Embryoid body formation by hanging drop --- p.90 / Chapter 3.5.5.2 --- Retinoic acid induced differentiation --- p.92 / Chapter Chapter IV --- Discussion / Chapter 4.1 --- General Discussion --- p.96 / Chapter 4.1.1 --- Cancer immunotherapy and dendritic cells --- p.96 / Chapter 4.1.2 --- Dendritic vaccine and tumor antigen --- p.97 / Chapter 4.1.3 --- Induced pluripotent stem cell technology and dendritic cells --- p.98 / Chapter 4.1.4 --- Tumor antigen presentation and dendritic cells --- p.98 / Chapter 4.1.5 --- D122 and cancer immunotherapy --- p.99 / Chapter 4.1.6 --- Method to introduce transcription factors for reprogramming --- p.100 / Chapter 4.1.7 --- Kinetics of reprogramming --- p.101 / Chapter 4.1.8 --- Culture condition for reprogramming D122_reNanog --- p.102 / Chapter 4.1.9 --- Reprogramming efficiency --- p.103 / Chapter 4.1.10 --- Establishment of D122-iPS-lC --- p.103 / Chapter 4.1.11 --- Differentiation of D122-iPS-1C --- p.104 / Chapter 4.2 --- Future Work --- p.106 / Chapter 4.3 --- Conclusion --- p.107 / Chapter Chapter V --- Bibliography --- p.108 / Appendix --- p.124

No description available.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet. / Härtill 4 uppsatser.

Eighty-two patients with breast cancer, twenty-seven patients with nasopharyngeal carcinoma and sixty-five patients with rheumatoid arthritis in this study were selected based on voluntary, randomization and double blind grouping criteria. / In nasopharyngeal carcinoma patients, the decrease in percentage and the absolute count of T lymphocytes in the TCM group was significantly lower than those in the placebo group. Besides, the decrease of the absolute count of T helper and T suppressor in the TCM group was significantly lower than that in the placebo group (all p < 0.05). The decrease may be due to radiotherapy. However, there was no significant difference in plasma sIL-2R and soluble tumor necrosis factor receptor 2 (sTNFR2) between the TCM group and the placebo group. / In rheumatoid arthritis patients, there was no significant difference in plasma. C-reactive protein (CRP), in the percentage, absolute count, and the ratio of CD4+/CD8+/NK/B lymphocytes between the TCM group and the placebo group. / Results showed that the absolute count of T helper lymphocytes (CD4+), the ratio of T helper lymphocytes (CD4+)/T suppressor and cytotoxic lymphocytes (CD8+), and the percentage and the absolute count of B lymphocytes were significantly elevated in the patients with breast cancer after taking Yunzhi-Danshen capsules, while plasma soluble interleukin-2 receptor (sIL-2R) concentration was significantly decreased (all p < 0.05). / This study shows that the selected traditional Chinese medicine have determinable immunomodulatory effects in patients with cancer and rheumatoid arthritis. (Abstract shortened by UMI.) / Traditional Chinese medicine (TCM) has been used to treat chronic diseases and tumor allegedly by immunomodulatory mechanisms. Breast cancer and nasopharyngeal cancer are prevalent carcinoma diseases in Hong Kong. The immune system of such patients could be adversely affected during the course of conventional chemotherapy or radiotherapy. Rheumatoid arthritis is an inflammatory autoimmune disease of the joints. The aim of this study was to assess the immunomodulatory effects of TCM Yunzhi-Danshen in auxiliary treatment of both kinds of cancer patients, and Lingzhi (Ganoderma Lucidum)-San Miao San ( Atractylodes lancea, Phellodendron amurense and Achyranthes bidentata B1) in supplementation treatment of patients with rheumatoid arthritis. / by Bao Yixi. / "July 2005." / Adviser: Wai-Kei Lam. / Source: Dissertation Abstracts International, Volume: 67-01, Section: B, page: 0166. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (p. 150-167). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Apesar de existir novas técnicas e múltiplas alternativas terapêuticas para o câncer de próstata localmente avançado, esta enfermidade se constitui em um grande problema de saúde pública mundial, resultando em índices significativos de morbidade e mortalidade, gerando desta forma um desafio para urologistas e oncologistas. Existem múltiplas e bem sucedidas estratégias de tratamento da doença localizada, tais como: a prostatectomia radical, a radioterapia externa conformacional, a braquiterapia e a crioablação. Em contraste, o tratamento da doença metastática e localmente avançada, freqüentemente necessita da alguma forma de bloqueio hormonal. Não existe consenso em vários aspectos da terapia hormonal para tumores localmente avançados tais como: o tipo de bloqueio androgênico a ser usado, terapia hormonal precoce ou tardia, associação com outras modalidades terapêuticas e o uso de bloqueio intermitente. Foi realizada uma revisão crítica deste tipo de tratamento, bem como as indicações atuais de bloqueio hormonal nos tumores de próstata localmente avançado. Não existem estudos prospectivos e randomizados que comparem as diversas formas de tratamento cirúrgico versus radioterápico do câncer de próstata localmente avançado. A hormonioterapia adjuvante à prostatectomia radical, na doença localmente avançada, parece reduzir a progressão tumoral bioquímica, porém, não há estudo que evidencie melhora na sobrevida livre de metástase ou na sobrevida global. O bloqueio androgênico neoadjuvante à prostatectomia radical aumenta a proporção dos pacientes com doença órgão-confinada e margens cirúrgicas negativas, porém sem efeito nas taxas de falha bioquímica do tratamento. A terapia hormonal adjuvante à radioterapia em pacientes portadores de câncer de próstata localmente avançado oferece vantagens na sobrevida global. A terapia hormonal neoadjuvante à radioterapia, em estudos multicêntricos e randomizados, resulta em melhor controle local do tumor bem como prolonga a sobrevida doença-específica. Não há, porém evidência de melhora na sobrevida global. O tratamento por tempo prolongado com bloqueadores hormonais adjuvante à radioterapia mostrou-se superior em relação à sobrevida global e sobrevida livre de doença quando comparado a um período curto de bloqueio, principalmente em pacientes com tumores indiferenciados (Gleason 8-10). Os análogos LHRH, orquiectomia ou o dietilestilbestrol se mostraram como opções de monoterapia, igualmente eficazes, para os pacientes que iniciam terapia hormonal de primeira linha, no tratamento da doença localmente avançada. Não existe evidência que justifique o bloqueio androgênico máximo como terapia hormonal de primeira linha ao invés de monoterapia. Existem vantagens potenciais na qualidade de vida e nos custos do tratamento quando realizada a ablação intermitente, mas a sua eficácia a longo prazo necessita ser confirmada / Despite new techniques and multiple therapeutic alternatives, locally advanced prostate cancer is a serious public health problem, resulting in significant morbidity and mortality rates, that remains a great challenge for urologists and oncologists. Several therapeutic strategies to treat localized prostate cancer have been successful such as conformational external beam radiation therapy, brachytherapy and cryoablation. In contrast, treatment of metastatic and locally advanced tumors may often involve androgenic suppression. However, there are no consensus on several aspects of hormonal therapy for locally advanced tumors such as the type of antiandrogenic drug to be used, early versus delayed hormonal therapy, association with other therapeutic modalities and the use of intermittent blockade. We set out to critically review important aspects and current indications of hormonal blockade in the locally advanced prostate tumors. There are no prospective and randomized study that compares current forms of surgical treatment versus radiation therapy of locally advanced prostate cancer. After radical prostatectomy, adjuvant hormonal therapy in the locally advanced disease reduces biochemical failure rates, although no benefit has been shown regarding metastatic free survival or overall suvival. Neoadjuvant androgen blockade enhances the proportion of patients with organ-confined disease and negative surgical margins but no benefit is seen regarding biochemical free recurrence. Neoadjuvant hormonal therapy to the radiotherapy improves local tumor control as well as it prolongs the diseasespecific survival, although there are no survival advantage. Adjuvant hormonal therapy offers overall survival advantage in patients with locally advanced prostate cancer treated with radiotherapy Long term adjuvant hormonal blockade offers survival benefit for patients with high Gleason score (8-10). LHRH analogues, bilateral orquiectomy and dietilestilbestrol were shown are equally effective as adjuvant therapy for patients with locally disease advanced. There are evidences that maximum androgenic blockade are not more efficient than monotherapy. Potential quality of life and costs advantages of intermittent ablation could be considered an alternative treatment for this group of patient

According to the results, Pa-PDT showed inhibitory effect on MDA-MB-231 cells in vitro with an IC50 value of 0.5 muM at 24 h. Pa-PDT was demonstrated to activate intracellular mitogen activated protein kinases (MAPK) pathways via reactive oxygen species (ROS) production. Pa-PDT IS also believed to induce extracellular signal-regulated kinase (ERK)-mediated autophagy and endoplasmic reticulum stress. Pa-PDT in combination with Tamoxifen is demonstrated to exert a synergetic effect in inhibiting cancer growth. The combination treatment induces both intrinsic and extrinsic apoptosis. Regarding the direct cancer cell killing activity, two dimensional gel electrophoresis screening revealed that Pa-PDT regulates proteins which involve in human leukocyte antigen (HLA) class I-restricted antigen-processing machinery. This activation of antigen presentation was confirmed by Western blot analysis and immunostaining. Furthermore, a cross-presentation of antigen with HLA class I proteins and 70-kDa heat shock protein was found in Pa-PDT-treated cells, as shown by the fluorescent microscopic observation and immunoprecipitation assay. Moreover, the immunogenicity of breast cancer cells was increased by Pa-PDT treatment that triggered phagocytic activity by human macrophages. Our findings provide the first evidence that Pa-PDT can trigger both apoptosis and anti-tumour immunity. / Cancer is one of the most lethal diseases worldwide. Treatments of cancer comprise surgical intervention, radiotherapy or chemotherapy; however, their side effects are still need to be overcome. In order to search for anti-cancer treatments with milder side effects and higher efficiency, traditional Chinese medicine (TCM) has been investigated. Previous study in our laboratory reported that pheophorbide a (Pa), an active compound purified from Scutellaria barbata, combined with photodynamic therapy (PDT) approach produces anti-tumour effect in a wide range of human cancers. Because of the lack of protocols for curing late phase breast cancer, my project is to investigate the therapeutic potential of Pa-PDT and its action mechanism on human breast cancer. A human breast cancer cell line MDA-MB-231, which is estrogen receptor nude and resistant to a conventional breast cancer drug tamoxifen, was used as an in vitro tumour model in my study to mimic the late stage of breast cancer. / Pheophorbide a (Pa) has been proposed to be a potential photosensitizer for the photodynamic therapy of human cancer. However, the immunomodulatory effect of Pa, in the absence of irradiation, has not yet been investigated. The present study revealed that Pa possessed immunostimulating effect on a murine macrophages cell line RAW 264.7. Pa could stimulate the growth of RAW 264.7 cells with the maximal effect at 0.5 muM after 48 h of treatment, where MAPK family including c-Jun N-tenninal kinase (JNK), ERK and p38 MAPK were activated by Pa treatment in a dose-dependent manner. Moreover, the induction of interleukin-6 and tumour necrosis factor-a secretion, and the enhancement of phagocytic activity were observed in Pa-treated RAW 264.7 cells. The results were similar in Pa-treated human immune competent cells (e.g. CD4+ and CD14+ cells) at higher Pa concentrations (from 1 to 10 muM). The present work is the first report to demonstrate the potential immunomodulatory effects of Pa on immune competent cells, apart from its well-known anti-tumour activity. / Bui Xuan, Ngoc Ha. / "December 2010." / Advisers: Fung Kwok Pui; Wong Chun Kwok. / Source: Dissertation Abstracts International, Volume: 73-04, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 123-144). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.