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Effects of CpG oligodeoxynucleotide on transcription factors GATA-3 and T-bet mRNA expression in asthmatic mice

Acta Pharmacologica Sinica 2005年第9期26卷 1117-1122页

作者： Zu-qun WU~(1,2) Yi-ping XU~3 Hua XIANG~4 Hua-hao SHEN~11 department of respiratory medicine,the second Affiliated hospital,4 department of Pathology,the First Affiliated hospital,School of medicine,Zhejiang university,Hangzhou 310009,china 3 department of respiratory medicine,renji hospital,shanghai second medical university shanghai 200001,china department of respiratory medicine the Second Affiliated Hospital Hangzhou China department of respiratory medicine Renji Hospital Shanghai Second Medical University Shanghai China department of Pathology the First Affiliated Hospital School of Medicine Zhejiang University Hangzhou China

Aim:To investigate effects ofCpG oligod'eoxynucleotide(CpG ODN)on the mRNAexpression of transcription factors GATA binding protein 3(GATA-3)and T-boxexpressed in T cells(T-bet)in asthmatic ***:An asthmatic mousemodel was established and treated with CpG *** inflammatory cells andeosinophils in bronchoalveolar lavage fluid(BALF)were counted and inflamma-tory cell infiltration in lung tissue was ***-y and interleukin-4concentrations in BALF and splenocyte culture supernatants were detected us-ing an enzyme-linked immunosorbent *** factor GATA-3 and T-bet mRNA expression in splenocytes and lung tissue were detected by reversetranscription-polymerase chain ***:Total inflammatory cells andeosinophils in BALF were reduced in the CpG ODN-treated group compared withthe asthma group,and inflammatory cell infiltration in lung tissue was also signifi-cantly *** ODN treatment increased the interferon-γ concentrationbut decreased the interleukin-4 concentration in both BALF and splenocyte cul-ture ***-3 mRNA expression was reduced in both lung tissueand splenocytes in the CpG ODN-treated group,while the mRNA ratio ofT-bet toGATA-3 in splenocytes was ***:CpG ODN treatment inhibitsairway inflammatory cell infiltration and regulates interferon-y/interleukin-4 syn-thesis in asthmatic mice,possibly through a mechanism of downregulation ofGATA-3 mRNA expression in both lung tissue and splenocytes.

关键词： asthma inflammation CpG oligodeoxy-nucleotide GATA-3 T-bet mice

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Differential Expression of Three Hypoxia-inducible Factor-α Subunits in Pulmonary Arteries of Rat with Hypoxia-induced Hypertension

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Acta Biochimica et Biophysica Sinica 2005年第10期37卷 665-672页

作者： Qi-Fang LI Ai-Guo DAI department of respiratory medicine,Human Institute of Gerontology,Human Province Geriatric hospital,Changsha 410001,china department of Anesthesiology.renji hospital,shanghai second medical university,shanghai,china department of respiratory medicine Hunan Institute of Gerontology Hunan Province Geriatric HospitalChangsha 410001 China

Hypoxia inducible transcription factor (HIF)-1α plays an important role in the development ofhypoxic pulmonary hypertension,but little is known about HIF-2α and HIF-3α with respect to transcriptionalregulation by *** examine the expression patterns of all HIF-α subunits (HIF-α,HIF-2α andHIF-3α) in pulmonary arteries of rats undergoing systemic hypoxia,five groups of healthy male Wistarrats were exposed to normoxia (N) and hypoxia for 3 (H),7 (H),14 (Hand 21 (*** arterial pressure (mPAP),vessel morphometry and right ventricular hypertrophy index *** were inflation fixed for immunohistochemistry and in situ hybridization,and homogenizedfor Western blot. mPAP increased significantly after 7 d of hypoxia [(18.4±0.4) vs. (14.4±0.4) mmHg,Hvs.N],reached its peak after 14 d of hypoxia,then remained *** artery remodeling and rightventricular hypertrophy developed significantly after 14 d of *** normoxia,HIF-1α and HIF-3αstaining were slightly positive regarding mRNA levels.A substantial alteration of HIF-1α and HIF-3α stainingoccurred in puhnonary arteries after 14 d and 7 d of hypoxia,respectively,but HIF-2α staining showed aninversed trend after 14 d of *** levels of all HIF-α subunits except HIF-3α showed a markedincrease corresponding to the duration of hypoxia,which was obtained by Western *** study foundthat HIF-1α. HIF-2α and HIF-3α may not only confer different target genes,but also play key pathogeneticroles in hypoxic-induced pulmonary hypertension.

关键词： hypoxia inducible factor-α hypoxia hypertension lung

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Upregulation of heparin-binding growth-associated molecule after spinal cord injury in adult rats

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Acta Pharmacologica Sinica 2004年第5期25卷 69-74页

作者： Yan-ting WANG2, Shu HAN2, Kai-hua ZHANG2, Yu JIN3, Xiao-ming XU2,4, Pei-hua LU2,52department of Neurobiology, shanghai second medical university, shanghai 200025, china 3department of Surgery,Xinhua hospital Affiliated to shanghai second medical university 4Kentucky Spinal Cord Injury Research Center,department of Neurological Surgery, university of Louisville School of medicine, Louisville, KY, 40292, USA department of Neurobiology Shanghai Second Medical University 上海 200025

AIM: To investigate whether traumatic spinal cord injury (SCI) induces changes of gene expression of heparin-binding growth-associated molecule (HB-GAM). METHODS: In a spinal cord transection model, HB-GAM ex-pression and cellular localization were examined using Northern blot, RT-PCR, immunohistochemistry and immun-ofluorescence double-labeling methods. RESULTS: HB-GAM mRNA was significantly upregulated in spinal cordtissues rostral and caudal to the injury at 7 d after SCI. HB-GAM gene expression was markedly increased at 3 d,peaked at 7 d, and declined to the baseline level at 28 d post-injury. During its peak expression, HB-GAM was co-localized in astrocytes, oligodendrocytes, and neurons in spinal cord tissues within 7 mm from the site of spinaltransection. CONCLUSION: SCI induces HB-GAM expression at both mRNA and protein levels in areas close tothe injury. Both neurons and glial cells expressed HB-GAM implying that HB-GAM played a role in the process ofinjury and/or repair following SCI.

关键词： heparin-binding growth-associated molecule spinal cord injuries immunohistochemistry reversetranscriptase polymerase chain reaction astrocytes neurons

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Design,synthesis and antitumor evaluation of a new series of N-substi-tuted-thiourea derivatives

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Acta Pharmacologica Sinica 2006年第9期27卷 1259-1271页

作者： Jian LI~2 Jin-zhi TAN~2 Li-li CHEN~2 Jian ZHANG~2 Xu SHEN~(2,3)Chang-lin MEI~(4,6)Li-li FU~4 Li-ping LIN~5 Jian DING~5 Bing XIONG~2 Xi-shan XIONG~4 Hong LIU~(2,6)Xiao-min LUO~(2,6)Hua-liang JIANG~(2,3)2 Drug Discovery and Design Centre,State Key Laboratory of Drug Research,shanghai Institute of Materia Medica,shanghai Institutes for Biological Sciences,Chinese Academy of Sciences,shanghai 201203.china 3 School of Pharmacy.East china university of Science and Technology,shanghai 200237,china 4 Division of Nephrology,department of Internal medicine,Changzheng hospital,second Military medical university,shanghai 200003,china 5 Division of Antitumor Pharmacology,shanghai Institute of Materia Medica,shanghai 201203.china Drug Discovery and Design Centre State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China School of Pharmacy East China University of Science and Technology Shanghai China Division of Nephrology Department of Internal Medicine Changzheng Hospital Second Military Medical University Shanghai China Division of Antitumor Pharmacology Shanghai Institute of Materia Medica Shanghai China

Aim:To design and synthesize a novel class of protein tyrosine kinase inhibitors,featuring the N-(2-oxo-1,2-dihydroquinolin-3-yl-methyl)-thiourea ***:First,compounds 1 and 2 were identified using the virtual screeningapproach in conjunction with binding assay based on surface plasmon ***,3 regions of compounds 1 and 2 were selected for chemical *** compounds were characterized potent inhibitory activities toward thehuman lung adenocarcinoma cell line SPAC ***:Forty new compounds(1-2,3a-g,4a-w,and 5a-l)were designed,synthesized and *** com-pounds(1,3e,41,4w,5a,and 5b)were found to show promising inhibitory activityagainst the SPAC1 tumor cell *** inhibitory activity of compound 5aincreases approximately 10 times more than that of the original compound ***:This study provides a promising new template with potential antitu-mot activity.

关键词： N-substituted-thiourea derivatives antitumor SPAC1 tyrosine kinase inhibitor virtual screening

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The expression and antigenicity identification of recombinant rat TGF-β1 in bacteria

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Cell Research 2001年第2期11卷 95-100页

作者： GAO CHUN FANG, XIAN TAO KONG, AXEL M GRESSNER, RALF WEISKIRCHEN department of Laboratory medicine, Changzheng hospital, second Military medical university, shanghai 200003, china Institute of Clinical Chemistry and Pathobiochemistry, Central Laboratory, R department of Laboratory medicine Changzheng Hospital Second Military Medical University Shanghai China Institute of Clinical Chemistry and Pathobiochemistry Central Laboratory RWTH-University Hospital Aachen Germany

In order to study structure-function details of TGF-β1, the recombinant mature form of rat TGF-β1 was expressed in bacteria. Synthesis of the 112 amino-acid carboxyl-terminal part of TGF-β1 (amino acid 279-390) was controlled by an inducible gene expression system based on bacteriophage T7 RNA polymerase. This system allowed an active and selective synthesis of recombinant TGF-β1. The molecular weight of expressed TGF-β1 monomer determined on SDS-polyacrylamide gel under reducing conditions was about 13 kD. Serial detergent washes combined with a single gel-filtration purification step were sufficient to purify the expression product to homogeneity. Amino-terminal sequencing revealed that the N-terminal of the recombinant protein was identical to the published data. In Western blot analysis the recombinant polypeptide showed excellent antigenicity against polyclonal TGF-β1 antibody. The mature recombinant rat TGF-β1 expressed in this study provides a useful tool for future detailed structural and functional studies.

关键词： TGF-β1 recombinant expression inclusion bodies purification antigenicity.

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Human C-C chemokine receptor 3 monoclonal antibody inhibits pulmonary inflammation in allergic mice

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Acta Pharmacologica Sinica 2007年第11期28卷 1791-1796页

作者： Kai WANG~2,Hua-hao SHEN~2,Wen LI,Hua-qiong HUANGdepartment of respiratory medicine,the second Affiliated hospital,medical School of Zhejiang university,Hangzhou 310009,china department of respiratory medicine the Second Affiliated Hospital Medical School of Zhejiang University Hangzhou China

Aim:To evaluate the effect of C-C chemokine receptor 3 (CCR3) blockade onpulmonary inflammation and mucus production in allergic ***:Weused the synthetic peptide of the CCR3 NH2-terminal as the immunizing antigenand generated murine monoclonal antibody against the human *** addition,the generated antibody was administered to mice sensitized and challenged *** inflammatory cells in bronchoalveolar lavage,cytokine levels,pulmonary histopathology,and mucus secretion were ***:TheWestern blotting analysis indicated that the generated antibody bound to *** allergic mice treated with the antihuman CCR3 antibody exhib-ited a significant reduction of pulmonary inflammation accompanied with the al-teration of ***:The antibody we generated was specific *** inhibition of airway inflammation and mucus overproduction by theantibody suggested that the blockade of CCR3 is an appealing therapeutical tar-get for *** present research may provide an experimental basis for thefurther study of this agent.

关键词： asthma C-C chemokine interleukin-4 lungs inflammation

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Therapeutic Angiogenesis:The Combination Point of Chinese and Western medicine in Prevention and Treatment of Ischemic Heart Disease

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Chinese Journal of Integrative medicine 2009年第5期15卷 323-324页

作者：吴宗贵 department of Cardiology Changzheng HospitalSecond Military Medical University

At present, the conventional medicine and revascularization （including interventional treatment and coronary bypass surgery） in ischemic heart disease have limited the clinical efficacy on the progress of the left ventricular remodeling and heart failure after myocardial infarction, and the mortality accounts about 50% of cardiovascular diseases. With the potential role of repairing injured or necrotic myocardium and improving the blood supply in ischemic region, heart cell therapy and therapeutic angiogenesis have become new highlights in disease treatment and the field of cardiovascular new strategies for the prevention and treatment of complications of endstage ischemic heart diseases.

关键词： Therapeutic Angiogenesis

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Murine fertilized ovum, blastomere and morula cells lacking SP phenotype

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Science china(Life Sciences) 2007年第6期50卷 762-765页

作者： XU YiXin1, HE ZhiYing2, ZHU HaiYing2, CHEN XueSong2, LI JianXiu2, ZHANG HongXia2, PAN XingHua3 & HU YiPing2 1 ChangZheng hospital, affiliated to second Military medical university, shanghai 200003, china 2 department of Cell Biology, second Military medical university, shanghai 200433, china 3 department of Genetics, Yale university School of medicine, New Haven, CT 06519, USA ChangZheng hospital affiliated to Second Military Medical University Shanghai China department of Cell Biology Second Military Medical University Shanghai China department of Genetics Yale University School of Medicine New Haven USA

In the field of stem cell research, SP (side population) phenotype is used to define the property that cells maintain a high efflux capability for some fluorescent dye, such as Hoechst 33342. Recently, many researches proposed that SP phenotype is a phenotype shared by some stem cells; some progenitor cells,; that SP phenotype is regarded as a candidate purification marker for stem cells. In this research, murine fertilized ova (including conjugate; single nucleus fertilized ova), 2-cell stage; 8-cell stage blastomeres, morulas; blastocysts were isolated; directly stained by Hoechst 33342 dye. The results show that fertilized ovum, blastomere; morula cells do not demonstrate any ability to efflux the dye. However, the inner cell mass (ICM) cells of blastocyst exhibit SP phenotype, which is consistent with the result of embryonic stem cells (ESCs) in vitro. These results indicate that the SP phenotype of ICM-derived ESCs is an intrinsic property; independent of the culture condition in vitro,; that SP phenotype is one of the characteristics of at least some pluripotent stem cells, but is not shared by totipotent stem cells. In addition, the result that the SP phenotype of ICM cells disappeared when the inhibitor verapamil was added into medium implies that the SP phenotype is directly associated with ABCG2. These results suggest that not all the stem cells demonstrate SP phenotype,; that SP phenotype might act as a purification marker for partial stem cells such as some pluripotent embryonic stem cells; multipotent adult stem cells, but not for all stem cells exampled by the totipotent stem cells in the very early stage of mouse embryos.

关键词： ABCG2/BCRP, stem cell, fertilized ovum, blastocyst, inner cell mass, embryonic stem cell, SP phenotype

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Onset time of complications in patients with severe acute pancreatitis receiving nonoperative therapy

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Hepatobiliary & Pancreatic Diseases International 2002年第1期1卷 143-145页

作者： Zeng-Yi Gong Yao-Qing Tang From the department of Surgery, Putuo District People’s hospital, shanghai 200060, china department of Surgery, Ruijin hospital, shanghai second medical university, shanghai 200025 china department of Surgery Putuo District Poeple's Hospital Shanghai 200060 China department of Surgery Ruijin Hospital Shanghai Second Medical Univ. Shanghai 200025 China

Objective: To retrospectively analyze 85 hospitalizednonoperative patients with severe acute pancreatitis(SAP) and to find out the stages of the ***: We statistically calculated the time of onsetof complications in these patients from 1987 to ***: The 95% confidence interval of total averagefor the complications of acute respiratory distress syn-drome (ARDS), shock and kidney failure was between2 to 4 days, and for encephalopathy, hemorrhage ofthe digestive tract, bacterial infection, fungous infec-tion and abscess between 3 to 6 days, 3 to 5 days, 13to 16 days, 13 to 16 days, and 14 to 23 days respec-tively. The 95% confidence interval of total averagein 18 deaths (21%) was between 5 to 6 days. ARDS,kidney failure, and shock occurred within 4 days, en-cephalopathy within 6 days (average 4.8±0.9 days),abscesses after 14 days, systemic bacterial infectionand fungous infection within half a month (average14.6±1.1days, 14.8±0.9days respectively), and deathwithin 6 ***: According to the time of the occurrence ofcomplications, we divide the courses of the disease intothree stages: early phase(first 4 days) with ARDS,kidney failure, shock, encephalopathy and hemorrhageof the digestive tract; middle phase (5-15 days) withbacterial infection and fungous infection; late phase(15 days after the onset) with abscess.

关键词： severe acute pancreatitis complication phase nonoperation

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Inhibition of hepatitis B virus by oxymatrine in vivo

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World Journal of Gastroenterology 2001年第1期7卷 49-52页

作者： Xiao Song Chen1 Guo Jun Wang1 Xiong Cai1 Hong Yu Yu2 Yi Ping Hu3 1department of Infectious Diseases, Changzheng hospital, the second Military medical university, shanghai 200003, china2department of Pathology, 3department of Cell Biology, department of Basic medicine, the second Military medical university, shanghai 200433, china department of Cell Biology the Second Military Medical University Shanghai 200433China. Yphu@***

AIM To investigate the anti-HBV effect ofoxymatrine (oxy) in *** HBV transgenic mice were producedby micro-injection of a 4.2kb fragmentcontaining the complete HBV *** level of HBsAg and HBcAg in thetransgenic mice liver was determined byimmunohistochemical *** Four groups (6 mice in each group)were injected intraperitoneally with oxy at thedosage of 100,200, and 300 mg/kg or with salineonce a day for 30 days. Both HBsAg and HBcAgwere positive in livers of all the six mice in thecontrol group (injected with saline), and werepositive in livers of two mice in 100 mg/kg groupand 300mg/kg group. In 200mg/kg group,HBsAg and HBcAg were negative in livers of allthe six mice. Based on the results, 200 mg/kg isthe ideal dosage to explore the effect of oxy atdifferent time points. According to the oxytreatment time, mice were divided into fourgroups: 10 d, 20 d, 30 d and 60 d (4 mice in eachgroup). Each mouse underwent liver biopsy twoweeks before the treatment of oxy. Down-regulation of HBsAg and HBcAg appeared aftertreatment of oxymatrine for 10 d and 20 d, Dane-like particles disappeared after the treatment ofoxy for 20d under electron microscopy,however, the expression level of HBsAg andHBcAg returned to normal 60 d later after *** oxymatrine can reduce thecontents of HBsAg and HBcAg in transgenic miceliver, longer treatment time and larger dosagedo not yield better effects.

关键词： Alkaloids Animals Antiviral Agents DNA, Viral Dose-Response Relationship, Drug Gene Expression Regulation, Viral Hepatitis B Hepatitis B Core Antigens Hepatitis B Surface Antigens Hepatitis B virus development Mice Mice, Transgenic Research Support, Non-U.S. Gov't Virus Replication

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