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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 departmentofrespiratorymedicine theSecondAffiliatedHospitalZhejiangUniversityHangzhou310009China

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

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Acta Pharmacologica Sinica 2004年第05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 Shanghai 200025 China

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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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 departmentofrespiratorymedicine HunanInstituteofGerontologyHunanProvinceGeriatricHospitalChangsha410001China

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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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 1department of Laboratory medicine Changzheng Hospital Second Military Medical University Shanghai 200003 China 1department of Laboratory medicine Changzheng Hospital Second Military Medical University Shanghai 200003 China 2Institute of Clinical Chemistry and Pathobiochemistry Central Laboratory RWTH-University Hospital Pauwelsstr 30 D-52074 Aachen Germany 2Institute of Clinical Chemistry and Pathobiochemistry Central Laboratory RWTH-University Hospital Pauwelsstr 30 D-52074 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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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 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

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 People's Hospital Shanghai 200060 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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Effects of Chinese Jianpi herbs on cell apoptosis and related gene expression in human gastric cancer grafted onto nude mice

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World Journal of Gastroenterology 2002年第5期8卷 792-796页

作者： Ai-Guang Zhao Hai-Lei Zhao Jin-Kun Yang Lai-Di Tang department of Oncology,Longhua hospital,shanghai university of Traditional Chinese medicine,shanghai 200032,china Xiao-Jie Jin renji hospital,shanghai second medical university,shanghai 200032,china department of Oncology Longhua Hospital Shanghai University of Traditional Chinese Medicine Shanghai 200032 China. aiguang@***

AIM: To explore the mechanism of the Sijunzi decoction and another Chinese herbal recipe (SRRS) based mainly on the Sijunzi decoction in treatment of gastric ***: A human gastric adenocarcinoma cell line SGC7901 grafted onto nude mouse was used as the animal model. The mice were divided into 3 groups, one control and the two representative experimental conditions. Ahimals in the two experimental groups received either Sijunzi decoction or SRRS over a 40-day period starting at 1st day after grafting. Control animals received saline on an identical schedule. Animals were killed 41 days after being *** effect of therapy was assessed by two ways: (1)tumor size was periodically measured during the life of the animals; (2) tumor weight was determined by a electron balance immediately after the animals killed. For detection of apoptotic cells, apoptotic indices(AI) were examined by the terminal deoxynucleotidyl transferase-mediated deoxyuddine tdphosphate fluorescence nick end labeling (TUNEL) *** alterations were observed with electron microscopy. S-P immunohistochemical method was used to detect the expression of Ki-67 in xenografts. Expression of bcl-2 and p53 was semiquantitatively detected using a reverse transcriptase-polymerase chain reaction (RT-PCR)***: When compared with controls, tumor growth (size and weight) was significantly inhibited by treatment with the Sijunzi decoction (P＜0.05) or SRRS (P＜0.01). The tumor inhibitory rate in the Sijunzi decoction group was 34.33 % and SRRS group 46.53 %. AI of human gastric cancer xenografts in nude mice was significantly increased to 16.24±3.21% using TUNEL method and 11.38±6.46 % by FACScan in the Sijunzi decoction group compared with the controls (TUNEL: 2.63±1.03 %, P＜0.01; FACScan: 7.15± 1.32 %, P＜0.05). SRRS group was also found a significantly increased AI by using TUNEL method and flow cytometry analysis compared with the controls (TUNEL: 13.18±3.05 %,P＜0.05; FACSc

关键词：胃癌健脾中药作用机制细胞凋亡基因表达动物实验

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3^H-脱氧葡萄糖掺入判断高能X线照射A549肺腺癌多细胞肿瘤球体的疗效

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复旦学报（医学版） 2003年第4期30卷 333-337页

作者：史德刚黄钢苗积生林祥通贺晓东苏斌复旦大学附属华山医院核医学科上海200040 上海第二医科大学附属仁济医院核医学科上海200001 上海市肺科医院核医学科上海市肺科医院放疗科上海200433

目的用3H-脱氧葡萄糖(3H-DG)掺入判断6MV X线照射A549肺腺癌多细胞肿瘤球体(MTS)的疗效。方法 MTS分别作低剂量分割和单次照射,中等剂量单次照射。MTS消化成单个细胞后用流式细胞仪检测细胞周期和凋亡比率。96孔盘每孔1个MTS,加37 kBq ... 详细信息

目的用3H-脱氧葡萄糖(3H-DG)掺入判断6MV X线照射A549肺腺癌多细胞肿瘤球体(MTS)的疗效。方法 MTS分别作低剂量分割和单次照射,中等剂量单次照射。MTS消化成单个细胞后用流式细胞仪检测细胞周期和凋亡比率。96孔盘每孔1个MTS,加37 kBq 3H-DG后掺入60 min,吸出MTS抽滤后测量β计数。结果 2～8 Gy低剂量照射后未见明显细胞周期阻滞和凋亡,分割照射组MTS的摄取减低,4.84 Gy单次照射后的摄取明显高于其他各组(P30 Gy照射后,细胞G2-M期阻滞百分率增加,各照射组3H-DG摄取均小于对照组(P3H-DG的能力降低;15 Gy照射后第10天有活细胞增殖,25或30 Gy照射后第40天有活细胞增殖。结论用3H-DG可以监测高能X线照射A549肺腺癌MTS的疗效,不同剂量照射后球体细胞的再增殖时间对于设计放疗方案和安排18F-FDG显像时间有指导作用。

关键词：肺腺癌细胞肺癌 3^H-脱氧葡萄糖高能X线照射多细胞肿瘤球体疗效放射疗法

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A549肺腺癌多细胞球体^(32)P内照射和高能X线外照射后放射生物学效应

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复旦学报（医学版） 2004年第6期31卷 632-636页

作者：史德刚黄钢苗积生林祥通贺晓东苏斌复旦大学附属华山医院核医学科上海第二医科大学附属仁济医院核医学科上海200001 上海市肺科医院核医学科上海200433 上海市肺科医院放疗科上海200433

目的　探讨A5 4 9肺腺癌多细胞球体内照射合适的3 2 P吸收剂量 ,以便临床对肺癌实体瘤作间质治疗选择核素内照射吸收剂量时给予参考。方法　 96孔盘每孔一个球体 ,15Gy3 2 P内照射 2 4h后当天用直线加速器产生的 6MV高能X线作另外一组... 详细信息

目的　探讨A5 4 9肺腺癌多细胞球体内照射合适的3 2 P吸收剂量 ,以便临床对肺癌实体瘤作间质治疗选择核素内照射吸收剂量时给予参考。方法　 96孔盘每孔一个球体 ,15Gy3 2 P内照射 2 4h后当天用直线加速器产生的 6MV高能X线作另外一组球体的连续 15Gy外照射 ,剂量率 2Gy/min。照射结束后第 8和第 15d用3 H 脱氧葡萄糖 (3 H DG)和3 H胸腺嘧啶核苷 (3 H TdR)摄取实验 ,流式细胞仪、光镜和电镜观察。结果　① 15Gy内、外照射结束 8d后3 H DG摄取明显低于对照组 (P 3 H Tdr摄取低于对照组 (P 36 ;15d后 ,内、外照射组和对照组3 H DG摄取比较P >0 .0 5 ,照射组的3 H Tdr摄取低于对照组 (P 3 2

关键词：外照射内照射对照组摄取 ^32P ^3H-TdR 多细胞球体高能电镜观察差别

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DNA Vaccine of SARS-Cov S Gene Induces Antibody Response in Mice

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Acta Biochimica et Biophysica Sinica 2004年第1期36卷 37-41页

作者： Ping ZHAO, Jin-Shan KE, Zhao-Lin QIN, Hao REN, Lan-Juan ZHAO, Jian-Guo YU, Jun GAO, Shi-Ying ZHU, and Zhong-Tian QI (department of Microbiology, second Military medical university, shanghai 200433, china department of Infectious Diseases, PLA hospital No.88, Tai'an 271000, china ) department of Microbiology Second Military Medical University Shanghai 200433 China.

The spike (S) protein, a main surface antigen of SARS-coronavirus (SARS-CoV), is one of themost important antigen candidates for vaccine design. In the present study, three fragments of the truncated S protein were expressed in E. coli, and analyzed with pooled sera of convalescence phase of SARS patients. The full length S gene DNA vaccine was constructed and used to immunize BALB/c mice. The mouse serum IgG antibody against SARS-CoV was measured by ELISA with E. coli expressed truncated S protein or SARS-CoV lysate as diagnostic antigen. The results showed that all the three fragments of S protein expressed by E. coli was able to react with sera of SARS patients and the S gene DNA candidate vaccine could induce the production of specific IgG antibody against SARS-CoV efficiently in mice with seroconversion ratio of 75% after 3 times of immunization. These findings lay some foundations for further understanding the immunology of SARS-CoV and developing SARS vaccines.

关键词： severe acute respiratory syndrome-coronavirus (SARS-CoV) spike protein DNA vaccine antibody

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