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Acta Biochimica et Biophysica Sinica 2005年第2期37卷 97-106页

作者： Pei-Zhou JIANG Ming GAN Hua HUANG Xin-Ming SHEN Shuang WANG Kai-Tai YAO Cancer Research Institute.Southern medical university,guangzhou 510515,china department of Histology and Embryology.Southern medical university,guangzhou 510515,china department of parasitology,zhongshan medical college.sun yat-sen university,guangzhou 510080.china Cancer Research Institute Southern Medical UniversityGuangzhou 510515 China department of Histology and Embryology Southern Medical UniversityGuangzhou 510515 China department of parasitology Zhongshan Medical College SUN Yat-sen UniversityGuangzhou 510080 China

Nasopharyngeal carcinoma(NPC)is a malignancy with high incidence in Southern china andSouth-East *** studies indicate that chemical carcinogen promoters,such as 12-O-tetradecanoylphorbol-13-acetate(TPA),are important ... 详细信息

关键词： 12-O-tetradecanoyl-phorbol-13-acetate nasopharyngeal carcinoma proteomics apoptosis

Temporal Control of Cre Recombinase-mediated in Vitro DNA Recombination by Tet-on Gene Expression System

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Acta Biochimica et Biophysica Sinica 2005年第2期37卷 133-138页

作者： Zhong-Min GUO Kang XU Ying YUE Bing HUANG Xin-Yan DENG Nu-Qi ZHONG Xun HONG Xi-Gu CHEN Dong XIAO Center of Experimental Animals.sun yat-sen (zhongshan) university.guangzhou 510080.china The First Affiliated Hospital of sun yat-sen university.sun yat-sen university.Guang-hou 510080.china Center of Experimental Animals Sun Yat-Sen (Zhongshan) University Guangzhou 510080 China The First Affiliated Hospital of sun yat-sen university Sun Yat-Sen University Guangzhou 510080 China

Conditional gene expression and gene deletion are important experimental approaches forexamining the functions of particular gene products in mouse models,These strategies exploiting Cre-medi-ated site-specific DNA recombination have been incorporated into transgenic and gene-targeting proceduresto allow in vivo manipulation of DNA in embryonic stem cells(ES cells)or living *** Cre/lox Psystem has become widely used in conditional gene targeting,conditional gene repair and activation,induc-ible chromosome translocation,and chromosome *** this project,we have employed the univer-sal transgenic system and the liver-specific promoter system for tightly temporal and liver-specific control ofCre gene expression in mice that(1)integrates the advantages of the Tet-on gene expression system and Cre//ox P site-mediated gene activation,and(2)simplifies the scheme of animal crosses through a combination oftwo control elements in a single transgene.A liver-specific apoE promoter was inserted into the promotercloning site upstream of the rtTA cassette of pCore construct to generate the transgene construct pApoErtTA-tetO-Cre,followed by demonstrating stringent regulation of doxycycline(Dox)-induced Cre-mediated re-combination in the lox P-flanked transcription STOP cassette-modified BEL-7402 *** is to say,in theabsence of Dox,the Cre gene is not expressed and will not induce site-specific recombination between twolox P sites,whereas on exposure to Dox,the Cre gene will be expressed and the recombination will ***,these data indicate that the Tet-on gene expression system is able to successfully and stringentlycontrol Cre expression in vitro,which lays a solid foundation for efficient and spatio-temporal Cre geneactivation in transgenic mice.

关键词： conditional gene regulation Cre recombinase human apolipoprotein E gene(apoE)promoter Tet-on gene expression system universal transgenic system

Protective effect of membrane cofactor protein against complementdependent injury

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Acta Pharmacologica Sinica 2005年第8期26卷 987-991页

作者： Dong XU, Shou-jian HUANG~1, Jin-qun WANG, Chu-kun WU department of Pharmacology, sun yat-sen medical college of sun yat-sen university, guangzhou 510080, china department of Pharmacology Sun Yat-Sen Medical College of Sun Yat-Sen University Guangzhou China

Aim: To evaluate the protective role of membrane cofactor protein (MCP CD46) on complement-dependent injury. Methods: MCP was separated by ion exchange chromatography on a DEAE sephadex A-50 column from pig erythrocyte ghosts. Its protective effect was tested in models such as cobra venom factor (CVF)-induced platelet metamorphosis and aggregation, human serum-induced injury in isolated working guinea pig heart and reverse passive Arthus reaction. Results: MCP inhibited CVF-induced platelet metamorphosis with an IC50of 56.7 mg/L±2.6 mg/L, and prevented injury induced by activated complement in isolated working guinea pig hearts. In the rat model of reverse Arthus reaction, MCP relieved the skin lesions induced by immune complexes. Conclusion: MCP has a protective effect against complement-dependent injury.

关键词： CD46 antigen complement heart reverse Arthus reaction

Oridonin induces apoptosis via PI3K/Akt pathway in cervical carcinoma HeLa cell line

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Acta Pharmacologica Sinica 2007年第11期28卷 1819-1826页

作者： Hong-zhen HU~2,Yue-bo YANG~3,Xiang-dong XU~4,Hong-wei SHEN~5,Yi-min SHU~6,Zi REN~7,Xiao-mao LI~3,Hui-ming SHEN~3,Hai-tao ZENG~(3,8)~2 department of Gynecology and Obstetrics,Yijishan Hospital,Wannan medical college,Wuhu 241001,china ~3 department of Gynecologyand Obstetrics,the Third Affiliated Hospital,sun yat-sen university,guangzhou 510080.china ~4 department of Surgery,the First AffiliatedHospital,sun yat-sen university,guangzhou 510080.china ~2 department of Gynecology and Obstetrics,the First Affiliated Hospital,sunyat-sen university,guangzhou 510080.china ~6 department of Obstetrics and Gynecology,IVF Program,Stanford university medicalCenter,Palo Alto,CA 94304,USA ~7 department of Gynecology and Obstetrics,Guangdong Provincial People's Hospital,guangzhou510080.china department of Gynecology and Obstetrics Yijishan Hospital Wannan Medical College Wuhu China department of Gynecology and Obstetrics the Third Affiliated Hospital Sun Yat-sen University Guangzhou China department of Surgery the First Affiliated Hospital Sun Yat-sen University Guangzhou China department of Gynecology and Obstetrics the First Affiliated Hospital Sun Yat-sen University Guangzhou China department of Obstetrics and Gynecology IVF Program Stanford University Medical Center Palo Alto USA department of Gynecology and Obstetrics Guangdong Provincial People's Hospital Guangzhou China

Aim:To investigate the apoptosis-inducing effect of oridonin,a diterpenoid iso-lated from Rabdosia rubescens,in the human cervical carcinoma HeLa cell ***:A morphological analysis,nuclear condensation,and fragmentation ofchromatin were monitored using Hoechst 33342 *** viability was as-sessed using the 3-(4,5-dimethylthiazol-(2)-yl)-2,5-diphenyl tetrazolium bromide(MTT) *** apoptosis and the apoptosis-related activation in the HeLa cellline were evaluated by flow cytometry and Western ***:Oridoninsuppressed the proliferation of the HeLa cell line in a dose-and *** treatment downregulated the activation of protein kinase B(Akt),the expression of forkhead box class O (FOXO) transcription factor,andglycogen synthase kinase 3 (GSK3).Oridonin also induced the release of cyto-chrome c accompanied by the activation of caspase-3 and poly-adenosine diphos-phate-ribose polymerase *** addition,Z-D(OMe)-E(OMe)-V-D(OMe)-FMK (z-DEVD-fmk),an inhibitor of caspases,prevented caspase-3 activation andabrogated oridonin-induced cell ***,oridonin treatment of the HeLa cellline downregulated the expression of the inhibitor of the apoptosis ***:Our results showed that oridonin-induced apoptosis involved sev-eral molecular *** may suppress constitutively activated targetsof phosphatidylinositol 3-kinase (Akt,FOXO,and GSK3) in the HeLa cell line,inhibiting the proliferation and induction of caspase-dependent apoptosis.

关键词： cervical carcinoma oridonin apoptosis Akt

Non-invasive Prenatal Gene Diagnosis: Progress through Cell-free Fetal DNA and RNA in Maternal Plasma and Urine

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中山大学学报（医学科学版） 2008年第S2期29卷 140-142页

作者： GUO Xun-yang, GUO Yi-bin ( department of medical Genetics, zhongshan School of Medicine, sun yat-sen university, guangzhou 510080, china )

Non-invasive prenatal gene diagnosis has been developed rapidly in the recent years, and numerous medical researchers are focusing on it. Such techniques could not only achieve prenatal diagnosis accurately, but also prevent tangential illness in fetuses and thus, reduce the incidence of diseases. Moreover, it is non-invasive prenatal gene diagnosis that prevents potential threaten and danger to both mothers and fetuses. Therefore, it is welcomed by clinical gynecologist and obstetrian, researchers of medical genetics, and especially, pregnancies. This review article touches briefly on the advanced development of using cell-free DNA, RNA in maternal plasma and urine for non-invasive prenatal gene diagnosis.

关键词： non-invasive prenatal gene diagnosis cell-free fetal DNA and RNA DNA and RNA detection maternal urine maternal plasma

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Effect of curcumin on multidrug resistance in resistant human gastric carcinoma cell line SGC7901/VCR

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Acta Pharmacologica Sinica 2005年第8期26卷 1009-1016页

作者： Xiao-qing TANG~1, Hu BI~1, Jian-qiang FENG~2, Jian-guo CAO~3 ~1 department of Physiology, Nanhua university, Hengyang 421001, china ~2 department of Physiology, zhongshan medical college, sun yat sen university, guangzhou 510080, china ~3 Institute of Oncology, Nanhua university, Hengyang 421001, china department of Physiology Nanhua University Hengyang China department of Physiology Zhongshan Medical College Sun Yatsen University Guangzhou China Institute of Oncology Nanhua University Hengyang China

Aim: To investigate the reversal effects of curcumin on multidrug resistance （MDR） in a resistant human gastric carcinoma cell line. Methods: The cytotoxic effect of vincristine （VCR） was evaluated by MTT assay. The cell apoptosis induced by VCR was determined by propidium iodide （PI）-stained flow cytometry （FCM） and a morphological assay using acridine orange （AO）/ethidium bromide （EB） dual staining. P-glycoprotein （P-gp） function was demonstrated by the accumulation and efflux of rhodamine 123 （Rh123） using FCM. The expression of P-gp and the activation of caspase-3 were measured by FCM using fluorescein isothiocyanate （FITC）-conjugated anti-P-gp and anti-cleaved caspase-3 antibodies, respectively. Results: Curcumin, at concentrations of 5 μmol/L, 10 μmol/L, or 20 μmol/L, had no cytotoxic effect on a parent human gastric carcinoma cell line （SGC7901） or its VCR-resistant variant cell line （SGC7901/VCR）. The VCR-IC50value of the SGC7901/VCR cells was 45 times more than that of the SGC7901ceUs and the SGC7901/VCR cells showed apoptotic resistance to VCR. SGC7901/VCR cells treated with 5 μmol/L, 10μmol/L, or 20 μmol/L curcumin decreased the IC50value of VCR and promoted VCR-mediated apoptosis in a dose-dependent manner. Curcumin （10 μmol/L） increased Rh123 accumulation and inhibited the effiux of Rh123 in SGC7901/VCR cells, but did not change the accumulation and efflux of Rh123 in SGC7901 ceils. P-gp was overexpressed in SGC7901/VCR cells, whereas it was downregulated after a 24-h treatment with curcumin （10 μmol/L）. Resistant cells treated with 1 μmol/L VCR alone showed 77% lower levels of caspase-3 activation relative to SGC7901 cells, but the activation of caspase-3 in the resistant cell line increased by 44% when cells were treated with VCR in combination with curcumin. Conclusion: Curcumin can reverse the MDR of the human gastric carcinoma SGC7901/VCR cell line. This might be associated with decreased P-gp function and expression, and the promotion of caspase-3

关键词： curcumin multidrug resistance apoptosis P-glycoprotein caspase-3

Temporal and Tight Hepatitis C Virus Gene Activation in Cultured Human Hepatoma Cells Mediated by a Cell-Permeable Cre Recombinase

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Acta Biochimica et Biophysica Sinica 2004年第10期36卷 687-694页

作者： Dong XIAO*, Kang XU1, Ying YUE, Zhong-Min GUO, Bing HUANG, Xin-Yan DENG, Huan TANG2, and Xi-Gu CHEN* Center of Experimental Animals, zhongshan university, guangzhou 510080, china 1The first affiliated hospital of sun yat-sen university, zhongshan university, guangzhou 510080, china 2The Third Military medical university, Chongqing 400010, china Center of Experimental Animals Zhongshan UniversityGuangzhou 510080 China The first affiliated hospital of sun yat-sen university Zhongshan UniversityGuangzhou 510080 China The Third Military medical universityChongqing 400010 China

Conditional gene expression has greatly facilitated the examination of the functions of particular gene products. Using the Cre/lox P switching expression system, we plan to develop efficient conditional transgeneactivation of hepatitis C virus core protein (HCV-C) cDNA (nucleotide 342–914) in the transgenic mice to overcome “immune tolerance” formed during the embryonic period and “immune escape” against hepatitis virus antigen in our project. To use this system in vivo, the dormant transgenic construct, i.e., pApoE-SCS-EGFP-HCV-C, was generated using techniques of standard molecular biology. The liver- specific human apoE promoter was here used to target expression of genes of interest (EGFP and HCV-C) to murine liver. Prior to generating the transgenic mice, the availability of Cre/lox P system and construct functionality were successfully verified by a cell-free recombination system and via checking the expression of EGFP and HCV-C in the human hepatoma cells at the mRNA and protein levels. These results suggest that the Cre/lox P system could tightly control expression of EGFP and HCV-C in vitro, which laid a solid foundation to conditionally activate expression of target gene(s) in transgenic mice by Cre-mediated site-specific recombination.

关键词： hepatitis C virus Cre/lox P switching expression system cell-free and intracellular recombination systems cell-permeable Cre recombinase EGFP human hepatoma cells

PPAR gamma inhibits growth of rat hepatic stellate cells and TGF beta-induced connective tissue growth factor expression

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Acta Pharmacologica Sinica 2006年第6期27卷 715-723页

作者： Kai sun Qian WANG~2 Xiao-hui HUANG department of Hepatobiliary Surgery,the First Affiliated Hospital of sun yat-sen university,guangzhou 510080.china department of Hepatobiliary Surgery the First Affiliated Hospital of Sun Yat-Sen University Guangzhou China

Aim:To investigate the effect of peroxisome proliferator-activated receptor gamma(PPARγ)activation on the growth of rat hepatic stellate cells(HSC)and trans-forming growth factor beta(TGF-β)-induced connective tissue growth factor(CTGF)***:After being treated with various amounts of thePPARγ natural ligand 15-deoxy-A 12,14-prostaglandin J2(15-d-PGJ2)or syntheticligand GW7845,the status of HSC proliferation and apoptosis were detected byMTT assay and flow ***,HSC were treated with PPARγ-specific antagonist GW9662 prior to the addition of 15-d-PGJ2or GW7845 andwere subsequently stimulated with TGF-β*** mRNA and protein levels ofCTGF expression were detected by semi-quantitative RT-PCR and Western blot-ting *** changes in the HSC were observed by ***:15-d-PGJ2and GW7845 markedly inhibited HSC prolifera-tion and induced cell apoptosis in a dose-dependent ***,PPARγligands significantly suppressed TGF-β1-induced CTGF expression(at both tran-scriptional and post-transcriptional levels)in HSC,and the inhibitory effect wasdramatically,if not completely,abolished by pretreatment with GW9662,sug-gesting that the inhibition was indeed mediated by PPARγ.Moreover,morpho-logical observation revealed that PPARγ activation caused obvious changes ofHSC from activated to quiescent ***:The PPARγligand hasa potent inhibitory effect on the growth of HSC and TGF-β1-induced CTGFexpression,which makes it a potential antifibrotic candidate for the treatment andprevention of hepatic fibrosis.

关键词： PPAR gamma transforming growth factor beta connective tissue growth factor hepatic stellate cells

Proteomic analysis of differential protein expression in early process of pancreatic regeneration in pancreatectomized rats

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Acta Pharmacologica Sinica 2006年第5期27卷 568-578页

作者： Ming YANG~(2,4) Wei LIU~(3,4) Chun-you WANG~(2,5) Tao LIU~2 Feng ZHOU~2 Jing TAO~2 Yang WANG~3 Ming-tao LI~3 ~2department of General Surgery,Union Hospital,Tongji medical college,Huazhong university of Science and Technology,Wuhan 430022,china ~3Proteomics Laboratory,zhongshan medical college.sun yat-sen university,guangzhou 510089.china department of General Surgery Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China Proteomics Laboratory Zhongshan Medical College Sun Yat-sen University Guangzhou China

Aim:A broad-range proteomic approach was applied to investigate the complex-ity of the mechanisms involved in pancreatic regeneration for identification ofnew targets of diabetes treatment and potential markers of pancreatic stem ***:A regeneration pancreatic model was induced by 90% partial pancreate-ctomy (Px) in *** in the protein expression in regenerating rat pancreason the third day after Px,as compared with rats that received sham surgery,wereanalyzed by using 2-D gel electrophoresis (2-DE),mass spectrometry (MS),andmass ***:2-DE revealed 91 spots with at least 1.5-fold in-creases in expression at 3d after pancreatectomy and 53 differentially expressedproteins that were identified by peptide mass fingerprinting (PMF).These in-cluded cell growth-related,lipid and energy metabolism-related,protein and aminoacid metabolism-related proteins,and signal transduction ***,CK8,L-plastin,hnRNP A2/B1,and AGAT are associated with embryogenesis and celldifferentiation,and may be new potential pancreatic stem cells ***:The proteome profiling technique provided a broad-based and effec-tive approach for the rapid assimilation and identification of adaptive proteinchanges during pancreas regeneration induced by *** dataclarify the global proteome during the pancreatic proliferation and differentiationprocesses,which is important for better understanding of pancreatic regenerationand for discovering of protein biomarkers for pancreatic stem cells.

关键词： partial pancreatectomy regeneration metabolism stem cell pancreas proteomics

An adaptive spatial clustering method for automatic brain MR image segmentation

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Progress in Natural Science:Materials International 2009年第10期19卷 1373-1382页

作者： Jingdan Zhang,Daoqing Dai Center for Computer Vision and department of Mathematics,sun yat-sen(zhongshan) university,guangzhou 510275,china Center for Computer Vision and department of Mathematics Sun Yat-Sen (Zhongshan) University Guangzhou 510275 China

In this paper,an adaptive spatial clustering method is presented for automatic brain MR image segmentation,which is based on a competitive learning algorithm-self-organizing map(SOM).We use a pattern recognition approach in terms of feature generation and classifier ***,a multi-dimensional feature vector is constructed using local spatial ***,an adaptive spatial growing hierarchical SOM(ASGHSOM) is proposed as the classifier,which is an extension of SOM,fusing multi-scale segmentation with the competitive learning clustering algorithm to overcome the problem of overlapping grey-scale intensities on boundary ***,an adaptive spatial distance is integrated with ASGHSOM,in which local spatial information is considered in the clustering process to reduce the noise effiect and the classification *** proposed method is validated by extensive experiments using both simulated and real MR data with varying noise level,and is compared with the state-of-the-art algorithms.

关键词： Self-organizing map Growing hierarchical SOM Adaptive spatial distance Image segmentation MR images