published_or_final_version / Dentistry / Doctoral / Doctor of Philosophy

Thesis--Auburn University, 2009. / Abstract. Vita. Includes bibliographical references.

Thesis--Auburn University, 2009. / Abstract. Vita. Includes bibliographical references.

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.

Hoi, Wan Heng. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 212-245). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Cheung, Tsz Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 97-116). / Abstracts in English and Chinese. / Thesis/Assessment Committee Members --- p.ii / Declaration for Plagiarism and Copyright --- p.iii / Abstract --- p.iv / 摘要 --- p.vi / Acknowledgements --- p.viii / Table of Contents --- p.ix / List of Abbreviations --- p.xii / List of Figures --- p.xv / List of Tables --- p.xvi / Chapter CHAPTER TWO: --- Introduction / Chapter 1.1 --- Background Information / Chapter 1.1.1 --- Breast Cancer --- p.1 / Chapter 1.1.2 --- General Statistics of Breast Cancer Worldwide and in Hong Kong --- p.1 / Chapter 1.1.3 --- Risk Factors for Breast Cancer --- p.2 / Chapter 1.1.4 --- Breast Cancer Treatment and Side Effects --- p.2 / Chapter 1.1.5 --- Types of Breast Cancer --- p.3 / Chapter 1.2 --- Estrogen and Estrogen Receptor / Chapter 1.2.1 --- Estrogen --- p.4 / Chapter 1.2.2 --- Estrogen Receptor --- p.5 / Chapter 1.2.3 --- Estrogen Receptor mediated Gene Transcription --- p.5 / Chapter 1.2.4 --- Estrogen Receptor Alpha and Estrogen Receptor Beta --- p.6 / Chapter 1.2.5 --- Estrogen Receptor Positive Breast Cancer and Treatment --- p.7 / Chapter 1.3 --- Estrogen metabolism and Cytochrome P450 family 1 (CYP1) members / Chapter 1.3.1 --- Estrogen Metabolism in Human --- p.9 / Chapter 1.3.2 --- CYP1A1 and CYP1B1 --- p.9 / Chapter 1.3.3 --- Estrogen Metabolism in Breast --- p.10 / Chapter 1.3.4 --- Carcinogenesis of Estrogens and Estrogen Metabolites --- p.13 / Chapter 1.3.5 --- The Importance of CYP1B1 in Carcinogenesis --- p.15 / Chapter 1.4 --- Aryl Hydrocarbon Receptor / Chapter 1.4.1 --- General Information of Aryl Hydrocarbon Receptor --- p.16 / Chapter 1.4.2 --- Signaling/Regulation Pathways of Aryl Hydrocarbon Receptor --- p.17 / Chapter 1.4.3 --- Crosstalk with Estrogen Receptor --- p.17 / Chapter 1.5 --- Introduction of Herba Scutellaria Barbata and its active ingredient Pheophorbide a --- p.19 / Chapter 1.6 --- Hyposthesis and Objectives --- p.21 / Chapter CHAPTER TWO: --- Direct Cytotoxic/Cytostatic Effect of Pheophorbide a / Chapter 2.1 --- Backgrounds --- p.22 / Chapter 2.2 --- Materials / Chapter 2.2.1 --- Chemicals --- p.24 / Chapter 2.2.2 --- Cell Lines --- p.26 / Chapter 2.2.3 --- "Cell Culture Mediums, Buffers and Consumables" / Chapter 2.2.3.1 --- Roswell Park Memorial Institute Tissue Culture Medium1640 (RPMI1640) --- p.26 / Chapter 2.2.3.2 --- RPMI 1640 (Phenol Red-free) --- p.26 / Chapter 2.2.3.3 --- Serum supplement - Fetal Bovine Serum (FBS) --- p.27 / Chapter 2.2.3.4 --- Serum supplement - Charcoal/Dextran Stripped FBS --- p.27 / Chapter 2.2.3.5 --- Antibiotics - Penicillin-Streptomycin (P/S) --- p.27 / Chapter 2.2.3.6 --- Trypsin (0.25%) with EDTA --- p.27 / Chapter 2.2.3.7 --- Trypsin (2.5%) (Phenol Red-free) with EDTA --- p.28 / Chapter 2.2.3.8 --- Dulbeccóةs Phosphate-Buffered Saline (D-PBS) --- p.28 / Chapter 2.2.3.9 --- Tissue Culture Flasks and Multi-well Plate --- p.28 / Chapter 2.2.3.10 --- Trypan Blue Solution --- p.29 / Chapter 2.2.4 --- Reagents for Direct Cytotoxity Test / Chapter 2.2.4.1 --- MTT Assay --- p.29 / Chapter 2.2.4.2 --- Tritiated Thymidine Incorporation Assay --- p.29 / Chapter 2.3 --- Methods / Chapter 2.3.1 --- Cell Culture --- p.30 / Chapter 2.3.2 --- Direct Cytotoxicity/Cytostatic Test / Chapter 2.3.2.1 --- MTT Assay --- p.31 / Chapter 2.3.2.2 --- Tritiated Thymidine Incorporation Assay --- p.32 / Chapter 2.3.3 --- Statistical Analysis --- p.32 / Chapter 2.4 --- Results / Chapter 2.4.1 --- The Cytotoxic Effect of Pheophorbide a --- p.34 / Chapter 2.4.2 --- The Combine Effect of Pheophorbide a with 17-β Estradiol and Tamoxifen Citrate --- p.34 / Chapter 2.5 --- Discussions --- p.48 / Chapter CHAPTER THREE: --- Mechanistic Study of Pheophorbide a / Chapter 3.1 --- Backgrounds --- p.53 / Chapter 3.2 --- Materials / Chapter 3.2.1 --- Real time PCR / Chapter 3.2.1.1 --- General Chemicals and Equipments --- p.54 / Chapter 3.2.1.2 --- RNA isolation --- p.55 / Chapter 3.2.1.3 --- Reverse Transcription --- p.55 / Chapter 3.2.1.4 --- Real Time PCR --- p.56 / Chapter 3.2.2 --- Western Blotting / Chapter 3.2.2.1 --- Microsome Isolation --- p.58 / Chapter 3.2.2.2 --- Measurement of Protein Concentration --- p.58 / Chapter 3.2.2.3 --- Western Blotting --- p.58 / Chapter 3.2.3 --- Estrogen Metabolism Assay / Chapter 3.2.3.1 --- Chemicals --- p.59 / Chapter 3.2.3.2 --- Estrogen Metabolites Extraction --- p.60 / Chapter 3.2.3.3 --- Liquid Chromatography/Mass Spectrometry --- p.60 / Chapter 3.3 --- Methods / Chapter 3.3.1 --- Real time PCR / Chapter 3.3.1.1 --- Cell Culture --- p.61 / Chapter 3.3.1.2 --- RNA Isolation and Reverse Transcription --- p.61 / Chapter 3.3.1.3 --- Real Time PCR --- p.62 / Chapter 3.3.2 --- Western Blotting / Chapter 3.3.2.1 --- Cell Culture --- p.63 / Chapter 3.3.2.2 --- Microsome Isolation --- p.63 / Chapter 3.3.2.3 --- Measurement of Protein Concentration --- p.64 / Chapter 3.3.2.4 --- Western Blotting --- p.64 / Chapter 3.3.3 --- Estrogen Metabolism Assay / Chapter 3.3.3.1 --- Preparation of Calibration Standard --- p.65 / Chapter 3.3.3.2 --- Cell Culture --- p.66 / Chapter 3.3.3.3 --- Estrogen Metabolites Extraction --- p.66 / Chapter 3.3.3.4 --- Liquid Chromatography/Mass Spectrometry --- p.67 / Chapter 3.3.4 --- Statistical Analysis --- p.68 / Chapter 3.4 --- Results --- p.69 / Chapter 3.5 --- Discussions --- p.80 / Chapter CHAPTER FOUR: --- Overall Conclusion and Future Directions / Chapter 4.1 --- Significance of the Study --- p.87 / Chapter 4.2 --- Overall Conclusion --- p.87 / Chapter 4.3 --- Limitation and Difficulties of the Study --- p.89 / Chapter 4.4 --- Future Directions --- p.89 / Appendices / "Appendix I The Melting Curve of real time PCR for β-actin, CYP1A1 and CYP1B1" --- p.92 / Appendix II The Calibration Curve of BSA for Protein Concentration Measurement --- p.93 / Appendix III The Representative Peak of Estradiol Metabolite Standards with corresponding Retention Time --- p.94 / Appendix IV The Calibration Curve of Different Estrogen Metabolites for LC/MS --- p.95 / Appendix V The Accuracy and Precision of Quality Control of Estradiol Metabolites --- p.96 / Bibliography --- p.97

Both mRNA and protein expression levels of P-glycoprotein, one of the major factors involved in drug resistance, was decreased in Pa-treated R-HepG2 cells. The chemo-sensitivity of these MDR cells towards doxorubicin would be enhanced by pretreatment of Pa. / In the study, 35 TCMs with historical background in treating liver diseases were screened. S. barbata was chosen for intensive studies based on its significant anti-hepatoma activity. Using bioassay-guided purification approach, an active component, pheophorbide a (Pa) - a chlorophyll derivative, was isolated from Scutellaria barbata. / Motivated by the severe health hazards worldwide caused by liver cancer, and the pronounced side effects of some recent anti-hepatoma agents in clinical treatment, we have initiated a research project in screening safe and effective agents from Traditional Chinese Medicine (TCM) for the treatment of hepatoma. The main objective of this research is to define the in vitro and in vivo anti-proliferative activities and to identify the action mechanisms of a TCM, the aerial part of Scutellaria barbata , in human hepatoma cells (HepG2 and Hep3B cells). / Pa exhibited anti-proliferative effects on HepG2 and Hep3B cells, through cell-cycle arrest and apoptosis, with IC50 values being 12.5 and 25.7 muM respectively. However, Pa produced insignificant cytotoxic effect on WRL-68 cells, a normal hepatic cell line. Pa also caused cell death in R-HepG2 cells, a multi-drug resistant (MDR) cell line developed from HepG2 cells. Microarray analysis indicated that a hypothetical protein FLJ10803 was found to be down-regulated upon the treatment of Pa on HepG2 cells. The sub-cellular localization of FLJ10803 was demonstrated by over-expression of the GFP fusion protein in HepG2 cells. / The anti-tumor effects of Pa could be enhanced by photodynamic therapy (PDT) approach, presumably due to the rapid generation of reactive oxygen species in the drug-binding site. Pa-PDT showed potent cytotoxicity on hepatoma cell lines, HepG2 and Hep3B, with IC50 values being 0.4 and 1.5 muM, respectively. The antitumor effects were confirmed by studies using animal model, where Pa treatment (300mug/kg/day, s.c.) could significantly inhibit the growth of Hep3B cells in nude mice after PDT treatment in vivo. Fluorescent imaging showed that Pa was located at the mitochondria, and the induction of cell death was found to be initiated by the mitochondrial dependent apoptotic pathway. Results of 2D-gel analysis suggested that Pa-PDT activated an immune-marker expression pathway that results in an over expression of HLA class I proteinsin Pa-PDT treated HepG2 cells. / To conclude, Pa may be a candidate for further development into an anti-hepatomic agent for clinical application. / Tang, Ming Kuen. / "September 2007." / Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4742. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 227-243). / 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.

Darbo tikslas: pagaminti pusiau kietas vaisto formas su baikalinės kalpokės (Scutellaria baicalensis Georgi) ekstraktu ir atlikti jų biofarmacinį vertinimą. Darbo uždaviniai: 1. Išvystyti ir validuoti metodiką baikalinės kalpokės ekstrakto veikliųjų junginių (baikalino, baikaleino ir vogonino) kokybinei ir kiekybinei analizei. 2. Pagaminti pusiau kietas vaisto formas (kremus, gelius ir gelifikuotus kremus) su baikalinės kalpokės ekstraktu ir atlikti jų biofarmacinį vertinimą. 3. Ištirti baikalinės kalpokės veikliųjų junginių atsipalaidavimą iš sumodeliuotų pusiau kietų vaisto formų tyrimais in vitro. Metodai: Ekstraktas analizuotas pritaikant fenolinių junginių ir flavonoidų nustatymą spektrofotometriniais metodais, spektrofotometrinį metodą laisvųjų DPPH• radikalų surišimo aktyvumui pagal troloksą nustatyti, efektyviosios skysčių chromatografijos metodiką biologiškai aktyviems junginiams – baikalinui, baikaleinui ir vogoninui – nustatyti kokybiškai ir kiekybiškai. Pusiau kietų vaisto formų gamyba: gelifikuotos sistemos gaminamos, naudojant karbomerą 980 kaip gelifikuojančią medžiagą ir įterpiant baikalinės kalpokės ekstraktą ištirpintą 30 % etanolyje; kremai gaminami naudojant skirtingus pagrindus, sudarytus iš polietileno ir vazelino aliejaus mišinys (PLW) ir iš hidrogenizuoto polideceno, polietileno ir tokoferolio mišinio (PLW PAO E), sumaišant ir homogenizuojant sistemas automatine maišykle; gelifikuoti kremai gaminami paruošiant dvi skirtingas fazes, kurios vėliau... [toliau žr. visą tekstą] / Objective of work: To prepare and analyze semisolid preparations with baikal skullcap (Scutellaria baicalensis Georgi) extract and evaluate their quality. Main tasks: 1. To develop and validate a suitable method for main active compounds (baicalin, baicalein and wogonin) of baikal skullcaps extract quality and quantity analysis. 2. To formulate semisolid dosage forms (creams, gels and jellified creams) containing extract of baikal skullcap and evaluate their quality. 3. To determine the release of baikal skullcap active compounds in vitro, from designed semisolid dosage forms. Methods: The analysis of the extract is executed by using spectrophotometric methods for analysis of total flavonoids, total phenolic compounds measuring, DPPH• free radical scavenging activity measuring according to trolox and capillary HPLC. Formulation of semisolids: jellified systems are prepared using carbomer 980 as a jellifying agent and adding baikal skullcap extract which is dissolved in 30 % ethanol; creams are prepared by stirring and homogenizing with an unguator using different bases: a mixture of polyethylene and vaseline (PLW) and a mixture of polyalphaolefine, polyethylene and vitamin E (PLW PAO E); jellified creams are made from two different phases, later they are jellified with poloxamer 407. Quality of the systems is evaluated: by detecting viscosity, measuring pH values and by testing release of active compounds in vitro. Results: Total amount of flavonoids in ethanolic 30 %... [to full text]

Methods. The intestinal absorption and metabolism of W and OA as well as the potential interactions among B, Wand OA were investigated at in vitro, in situ and in vivo levels. Various models were employed including Caco-2 cell monolayer model, in vitro enzymatic kinetics study, rat in situ single-pass intestinal perfusion model and in vivo pharmacokinetic study in rats. / Purpose. Scutellariae baicalensis Georgi is a medicinal plant widely distributed in Asia. Its dried root, Radix Scutellariae (RS), has been extensively used in Chinese and Japanese medicine. Six flavones including baicalein (B), wogonin (W), oroxylin A (OA) and their corresponding glucuronic acid conjugates (BG, WG, OAG) are the major bioactive components in RS. Our previous studies on B revealed an extensive first-pass metabolism during its absorption. Hence, it is expected that W and OA which have the similar structures as B, may share similar absorption and metabolic pathways as B. The present project aims to (1) establish an assay method for better quality control of RS; (2) provide further biopharmaceutic characterizations ofW and OA in RS; (3) investigate the potential pharmacokinetic interactions among B, Wand OA. / Results. Similar to B, Wand OA showed favorable permeability in both the Caco-2 cell and the rat in situ single-pass perfusion models. However, they experienced extensive first-pass metabolism, mainly in the form of glucuronidation. Intracellularly formed WG and OAG could be effluxed to both the apical side (lumen side) and basolateral side (mesenteric blood side) mainly by MRPs, which was confirmed by inhibition transport studies in Caco-2 cells and transfected MDCK cells. The glucuronidation rate of OA was higher than that of W, which was observed by enzymatic kinetics studies by sub-cellular fractions with intrinsic clearances (Vmax/K m, mul/min/mg) of 456 to 4170 for W and 509∼5038 for OA. UGT 1A9 was the most potent metabolic enzyme for hepatic glucuronidation, while UGTs 1A8 and 1AlO were responsible for the intestinal glucuronidation of W and OA. The in vivo rat pharmacokinetics studies showed that W and OA may be readily absorbed and extensively metabolized with no parent compound detectable in blood after oral administration of W and OA. A new metabolite of W was identified to be the glucuronic acid conjugate at 5-0H of W. After co-administration of B, W and OA, decreased formation of BG, WG and OAG was observed in in vitro enzymatic kinetics study. Further studies in absorption models of Caco-2 cell monolayer and rat in situ single-pass intestinal perfusion demonstrated the enhancement in absorption of B, W and OA and decrease of BG, WG and OAG after the co-administration of B, W and OA. The ultimate pharmacokinetics interaction study revealed that glucuronides were the predominant form in systemic circulation and the AUC of OAG significantly increased after co-administration of B, Wand OA. Conclusion: Similar to B, Wand OA may be well absorbed followed by extensive first-pass metabolism, which was mediated by various UGT isozymes. During absorption, the intracellularly formed WG and OAG were mainly effluxed by MRPs to both the lumen and mesenteric blood side of the intestine. Both in vitro and in situ models indicated that interactions among B, W and OA would lead to decreased glucuronidation and increased absorption of parent flavones. Due to extensive metabolism in vivo, only glucuronides appeared in systemic circulation after co-administration of B, W and OA in rats. The resulted increased systemic exposure of OAG indicated that the co-administration might lead to the enhancement of bioavailability for the studied flavones in the form of glucuronides. / Li, Chenrui. / Adviser: Zuo Zhong. / Source: Dissertation Abstracts International, Volume: 73-03, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 201-236). / 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. / Abstract also in Chinese.

Yip Wai Nga. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 93-101). / Abstracts in English and Chinese. / Abstract (English version) --- p.i / Abstract (Chinese version) --- p.v / Acknowledgements --- p.viii / Table of contents --- p.ix / List of figures --- p.xii / List of abbreviation --- p.xv / Chapter Chapter I: --- Introduction --- p.1 / Chapter I.1 --- Transepithelial ion transport --- p.1 / Chapter I.1.1 --- Fluid secretion in colon --- p.1 / Chapter I.1.2. --- Cellular mechanism of chloride secretion --- p.3 / Chapter 1.2. --- Regulation of chloride secretion in T84 cells --- p.6 / Chapter I.2.1 --- Human colonic T84 cells as the study model --- p.6 / Chapter I.2.2 --- Signal transduction pathways of chloride secretion in T84 cells --- p.13 / Chapter I.3. --- Pharmacological actions of Scutellariae Radix --- p.13 / Chapter I.3.1. --- What is Scutellariae Radix? --- p.13 / Chapter I.3.2. --- Some biological and pharmacological actions of Scutellariae Radix --- p.13 / Chapter I.4. --- Effects of Scutellariae Radix and its major flavonoid baicalein on ion transport in T84 cells --- p.15 / Chapter I.4.1. --- Effects of Scutellariae Radix extract on ion transport in T84 cells --- p.15 / Chapter I.4.2. --- Biological effects of baicalein --- p.15 / Chapter I.5 --- "Relationship of Coptidis rhizoma and its active ingredient berberine, with Scutellariae radix in traditional remedies" --- p.18 / Chapter I.6 --- Aim of study --- p.20 / Chapter Chapter II: --- Methods and Materials --- p.21 / Chapter II.1. --- Culture technique of the T84 cells --- p.21 / Chapter II.2. --- Conventional short-circuit current (Isc) measurement --- p.24 / Chapter II.2.1. --- Experimental setup --- p.24 / Chapter II.2.2. --- Preparation of the permeable supports --- p.27 / Chapter II.2.3. --- Cell seeding --- p.27 / Chapter II.2.4. --- Short-circuit measurement --- p.29 / Chapter II.2.5 --- Short-circuit measurement in nystatin-permeabilized T84 monolayers --- p.30 / Chapter II.3. --- Measurement of protein kinase A activity --- p.31 / Chapter II.4. --- Solutions and chemicals --- p.32 / Chapter II.5. --- Statistical analysis --- p.33 / Chapter Chapter III: --- Result --- p.34 / Chapter III.1. --- Effect of SRE on transepithelial ion transport processes in T84 monolayers --- p.35 / Chapter III.1.1 --- Effect of SRE and baicalein on baseline Isc --- p.35 / Chapter III.1.2 --- Effect of ion channel blockers on SRE-stimulated Isc --- p.39 / Chapter III.1.3 --- Effect of K+ channel blockers on SRE-stimulated Isc --- p.42 / Chapter III.1.4 --- Effect of SRE in C1- free solution --- p.48 / Chapter III.2. --- Effect of SRE on apical C1- conductance and basolateral K+ conductance in nystatin-permeabilized T84 monolayers --- p.51 / Chapter III.2.1 --- Effect of SRE and baicalein on baseline IC1 --- p.51 / Chapter III.2.2 --- Study of apical C1- conductance in T84 monolayers --- p.54 / Chapter III.2.3 --- Interaction of SRE and forskolin --- p.61 / Chapter III.2.4 --- Study of basolateral K+ conductance in T84 monolayers --- p.62 / Chapter III.3 --- Effect of SRE and baicalein on PKA activities in T84 cells --- p.64 / Chapter III.3.1 --- Effect of Scutellariae Radix on PKA activity --- p.66 / Chapter III.3.2 --- Effect of baicalein on PKA activity --- p.69 / Chapter III.3.3. --- Effect of berberine on PKA activity --- p.69 / Chapter III.3.3. --- Interaction of baicalein and berberine on PKA activity --- p.74 / Chapter Chapter IV: --- Discussion --- p.77 / Chapter IV.1 --- "Scutellariae Radix，Coptidis Rhizoma, and gastrointestinal secretory function" --- p.77 / Chapter IV.2 --- SRE- and baicalein-induced increase in Isc --- p.79 / Chapter IV.3 --- Cellular signaling mechanisms underlying the effect of SRE and baicalein --- p.82 / Chapter IV.4 --- "Interaction between Scutellariae Radix and Coptidis Rhizoma - the ""ying and yang"" hypothesis" --- p.89 / Chapter IV.5 --- Summary --- p.91