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| 1 | A complete sequence and comparative analysis of a SARS-associated virus(Isolate BJ01)显示文摘The genome sequence of the Severe Acute Respiratory Syndrome (SARS)-associated virus provides essential information for the identification of pathogen(s), exploration of etiology and evolution, interpretation of transmission and pathogenesis, development of diagnostics, prevention by future vaccination, and treatment by developing new drugs. We report the complete genome sequence and comparative analysis of an isolate (BJ01) of the coronavirus that has been recognized as a pathogen for SARS. The genome is 29725 nt in size and has 11 ORFs (Open Reading Frames). It is composed of a stable region encoding an RNA-dependent RNA polymerase (composed of 2 ORFs) and a variable region representing 4 CDSs (coding sequences) for viral structural genes (the S, E, M, N proteins) and 5 PUPs (putative uncharacterized proteins). Its gene order is identical to that of other known coronaviruses. The sequence alignment with all known RNA viruses places this virus as a member in the family of Coronaviridae. Thirty putative substitutions have been identified by comparative analysis of the 5 SARS- associated virus genome sequences in GenBank. Fifteen of them lead to possible amino acid changes (non-synonymous mutations) in the proteins. Three amino acid changes, with predicted alteration of physical and chemical features, have been detected in the S protein that is postulated to beinvolved in the immunoreactions between the virus and its host. Two amino acid changes have been detected in the Mprotein, which could be related to viral envelope formation. Phylogenetic analysis suggests the possibility of non-human origin of the SARS-associated viruses but provides noevidence that they are man-made. Further efforts should focus on identifying the etiology of the SARS-associated virus and ruling out conclusively the existence of otherpossible SARS-related pathogen(s). | QIN E'de ZHU Qingyu YU Man FAN Baochang CHANG Guohui SI Bingyin YANG Bao PENG Wenming JIANG Tao LIU Bohua DENG Yongqiang LIU Hong ZHANG Yu WANG Cui LI Yuquan GAN Yonghua LI Xiaoyu L Fushuang TAN Gang CAO Wuchun, YANG Ruifu Institute of Microbiology and Epidemiology, Chinese Academy of Military Medical Sciences, Beijing 100071, China WANG Jian, LI Wei, XU Zuyuan, LI Yan, WU Qingfa, LIN Wei, CHEN Weijun, TANG Lin, DENG Yajun, HAN Yujun, LI Changfeng, LEI Meng, LI Guoqing, LI Wenjie, L Hong, SHI Jianping, TONG Zongzhong, ZHANG Feng, LI Songgang, LIU Bin, LIU Siqi, DONG Wei, WANG Jun, Gane K-S Wong, YU Jun & YANG Huanming* Beijing Genomics Institute, Chinese Academy of Sciences, Beijing 101300 National Center for Genome Information, Beijing 101300, China | 2003 | Chinese Science Bulletin2003，48，10： | 121 |
| 2 | Alterations in metastatic properties of hepatocellular carcinoma cell following H-ras oncogene transfection显示文摘AIM To demonstrate the relationship betweenH-ras oncogene and hepatocellular carcinoma(HCC) metastasis.METHODS Activated H-ras oncogene wastransfected into SMMC 7721, a cell line derivedfrom human HCC, by calcium phosphatetransfection method. Some metastasis-relatedparameters were detected in vitro, includingadhesion assay, migration assay, expression ofcollagenase ⅣV (c ⅣV ase) and epidermal growthfactor receptor (EGFR).RESULTS The abilities of H-ras-transfected cellclones in adhesion to laminin (LN) or fibronectin(FN), migration, c Ⅳ ase secretion increasedmarkedly, and the expression of EGFR elevatedmoderately. More importantly, these alterationswere consistent positively with the expressionof p21, the protein product of H-ras oncogene.CONCLUSION H-ras oncogene could inducethe metastatic phenotype of HCC cell in vitro toraise its metastatic potential. | Qing Wang~1 Zhi Ying Lin~2 Xiao Li Feng~3 ~1Department of Microbiology,Medical Center of Fudan University.the former Shanghai Medical University,Shanghai 200032,China ~2Liver Cancer Institute,Zhongshan Hospital,Shanghai 200032,China ~3Shanghai Institute of Biochemistry,Academy Sinica,Shanghai 200031,ChinaQing Wang earned master degree from Shanghai Medical University in 1996,now a senior lecturer of microbiology,specialized in the role of oncogcncs on tumor metastasis,having 8 papers published. | 2001 | World Journal of Gastroenterology2001，7，3： | 48 |
| 3 | Expression of vascular endothelial growth factor and its role in oncogenesis of human gastric carcinoma显示文摘AIM To establish the role of vascular endothelial growth factor (VEGF) in the oncogenesisof human gastric carcinoma more directly.METHODS The expression of VEGF and its receptor kinase-domain insert containing receptor (KDR) in human gastric cancer tissue were observed by immunohistochemical staining. VEGF levels were manipulated in human gastric cancer cell using eukaryotic expression constructs designed to express the complete VEGF165 complimentary DNA in either the sense or antisense orientation. The biological changes of the cells were observed in which VEGF was up-regulated or downregulated.RESULTS VEGF-positive rate was 50%, and VEGF was mainly localized in the cytoplasm and membrane of the tumor cells, while KDR was mainly located in the membrane of vascular endothelial cells in gastric cancer tissues and peri-cancerous tissue. In 2 cases of 50 specimens, the gastric cancer cells expressed KDR,localized in both the cytoplasm and membrane.Introduction of VEGF165 antisense into human gastric cancer cells ( SGC-7901, immunofluorescence intensity,31.6%)) resulted in a significant reduction in VEGFspecific messenger RNA and total and cell surface VEGF protein ( immunofluorescence intensity, 8.9%)(P<0.05). Conversely, stable integration of VEGF165 in the sense orientation resulted in an increase in cellular and cell surface VEGF (immunofluorescence intensity,75.4%) (P<0.05). Lowered VEGF levels were associated with a marked decrease in the growth of nude mouse xenografted tumor (at 33 days postimplantation, tomor volume: 345.40 ± 136.31 mm3) (P<0.05 vs control SGC7901 group: 1534.40 ± 362.88 mm3), whereas up-regulation of VEGF resulted in increased xenografted tumor size (at 33 days postimplantation, tomor volume: 2350.50 ± 637.70mm3) (P<0.05 vs control SGC-7901 group).CONCLUSION This study provides direct evidence that VEGF plays an important role in the oncogenesis of human gastric cancer. | Du-Hu Liu Xue-Yong Zhang Dai-Ming Fan Yu-Xin Huang Jin-Shan Zhang Wei-Quan Huang Yuan-Qiang Zhang Qing-Sheng Huang Wen-Yu Ma Yu-Bo Chai Ming Jin Institute of Digestive Disease,Xijing Hospital,~2 Department of Gastroenterology,Tangdu Hospital,~3Department of Histology and Embryology,~4 Department of Microbiology,~5 Department of Biochemistry,Fourth Military Medical University,Xi’an 710033,Shaanxi Province,China | 2001 | World Journal of Gastroenterology2001，7，4： | 37 |
| 4 | Formation and Water Stability of Aggregates in Red Soils as Affected by Organic Matter显示文摘FormationandWaterStabilityofAggregatesinRedSoilsasAffectedbyOrganicMatter￥ZHANGMINGKUI;HEZHENLI;CHENGUOCHAO;HUANGCHANGYONGIan... | ZHANG MINGKUI HE ZHENLI CHEN GUOCHAO HUANG CHANGYONGI andM. J. WILSON￣2(￣1DePartment of Land Use and Applied Chemistry, Zhejiang Agricultural University, Hangzhou 31O029(China))(￣2Soils and Soil Microbiology Division, Macaulay Land Use Research Inst | 1996 | Pedosphere1996，6，1： | 38 |
| 5 | Diversity and ecological distribution of endophytic fungi associated with medicinal plants显示文摘A total of 973 isolates of endophytic fungi were recovered from 1144 tissue fragments of the six me-dicinal plant species belonging to 4 families collected in the Beijing Botanical Garden. Of these isolates 778 sporulated and were identified into 21 taxa by morphological characteristics. Among the taxa 11 belonged to Coelomycetes, 6 to Ascomycetes, and 4 to Hyphomycetes. Various numbers of endophytic fungi (5―8 taxa) were obtained from each plant. Alternaria alternata was the dominant species in the 6 plants, and Microsphaeropsis conielloides was also dominant in Eucommia ulmoides. There were high colonization rates (47.9%―63.1%) and isolation rates (0.7―0.93) of endophytic fungi, and they were conspicuously higher in twigs than those in leaves in the 6 plants examined. The colonization and isolation rates of endophytic fungi increased with the twig age. The results based on the analyses of cluster and Sorenson's similarity coefficients indicated that some endophytic fungi showed a certain degree of host and tissue preference. | SUN JianQiu1,2,3, GUO LiangDong1, ZANG Wei2,3, PING WenXiang4 & CHI DeFu3 1 Systematic Mycology and Lichenology Laboratory, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China 2 College of Life Science and Engineering, Qiqihar University, Qiqihar 161006, China 3 College of Forestry, Northeast Forestry University, Harbin 150040, China 4 College of Life Science, Heilongjiang University, Harbin 150080, China | 2008 | Science China(Life Sciences)2008，51，8： | 34 |
| 6 | Specific CEA-producing colorectal carcinoma cell killing with recombinant adenoviral vector containing cytosine deaminase gene显示文摘瞄准:到杀死 CEA 积极颜色明确地使用 E 关口 i cytosine 脱氨基酶(CD ) 的表面的癌房间自杀基因,新复制缺乏的 recombinant adenoviral 向量在在哪个 CD,基因在 CEA 倡导者下面被控制被构造并且它的在里面 vitro 细胞毒素的效果被评估。方法:梭原生质标志包含 CD 基因和 CEA 基因的规章的顺序在 293 房间紧张与侵入人体气管粘膜的病菌染色体 DNA 的右手臂被构造并且重新结合。点弄污, PCR 被用来识别积极的匾。侵入人体气管粘膜的病菌的纯化被执行与在 CsCl 步坡度和滴定极端集中与匾形成试金被测量。细胞毒素的效果是有 MTT 方法的 assayed, 5-FC 的百分之五十抑制集中(IC (50 )) 用一个曲线试穿参数被计算。表面的癌房间线,是生产 CEA,和 CEA-nonproducing Hela 房间衬里的人的颜色在 cytological 测试被使用。确定的 recombinant 侵入人体气管粘膜的病菌向量 AdCMVCD, CD 基因在 CMV 倡导者下面在被控制,被用作病毒控制。量的结果被表示为平均数的吝啬的 +/- SD。统计分析用 ANOVA 测试被执行。结果:需要的 recombinant 侵入人体气管粘膜的病菌向量被称为 AdCEACD。点弄污和 PCR 的结果证明 recombinant 侵入人体气管粘膜的病菌包含了 CEA 倡导者和 CD 基因。病毒 titer 是大约 5.0 X 10 (14 ) pfu/L (在纯化以后的 -1) 。 生产CEA Lovo 房间对 5-FC 敏感并且与 AdCEACD 和 AdCMVCD 在感染以后有一样的细胞毒素的效果(在父母 Lovo 房间,感染 100 M.O.I AdCEACD 的 Lovo 房间和感染 10 M.O.I AdCMVCD 的 Lovo 房间的 5-FC 的 IC ( 50 )价值是 >15000 , 216.5+/-38.1 和 128.8+/-25.4 micromol.L (-1), P<0.001 ,分别地),并且当侵入人体气管粘膜的病菌的 m.o.i 被提高时, 5-FC 的 cytotoxicity 因此增加了( 5-FC 的 IC ( 50 )的价值被归结为 27.9+/-4.2 micromol.L (在 1000 M.O.I AdCEACD 的-1)感染了 Lovo 房间和 24.8+/-7.1 micromol.L (在 100 M.O.I AdCMVCD 的-1)感染了 Lovo 房间, P<0.05 , P<0.01 ,分别地)。CEA-nonproducing Hela 房间没与 AdCEACD 在感染以后有效果,但是 Hela 房间与 AdCMVCD 在感染以后有细胞毒素的敏感到 5-FC (在在 10 M.O.I 感染 AdCMVCD 的父母 Hele 房间和 Hela 房间的 5-FC 的 IC (50 ) 是 >15000 和 214.5+/-31.3 micromol.L (-1), P<0.001 ) 。AdCEACD/5-FC 系统也有旁观者效果,并且当 transfected 房间的比例仅仅是 10% 时,生存能力是大约 30% 。结论:recombinant 侵入人体气管粘膜的病菌向量 AdCEACD 有房间的特性类型特定的基因交货。AdCEACD/5-FC 系统可以变得一新,为 CEA 积极的瘤的基因治疗的有势力和特定的途径,特别结肠癌。 | Li-Zong Shen Wen-Xi Wu Qiang Ding Yi-Bing Hua,Department of General Surgery,The First Affiliated Hospital of Nanjing Medical University,Nanjing,210029,Jiangsu Province,China De-Hua Xu Zhong-Cheng Zheng Xin-Yuan Liu,Shanghai Institute of Biochemistry and Cell Biology,The Chinese Academy of Sciences,Shanghai,200031,China Kun Yao,Department of Microbiology and Immunology,Nanjing Medical University,Nanjing,210029,Jiangsu Province,China | 2002 | World Journal of Gastroenterology2002，8，2： | 29 |
| 7 | Production of proinflammatory cytokines in the human THP-1 monocyte cell line following induction by Tp0751,a recombinant protein of Treponema pallidum显示文摘The tissue destruction characteristic of syphilis infection may be caused by inflammation due to Treponema pallidum and the ensuing immune responses to the pathogen.T.pallidum membrane proteins are thought to be potent inducers of inflammation during the early stages of infection.However,the actual membrane proteins that induce inflammatory cytokine production are not known,nor are the molecular mechanisms responsible for triggering and sustaining the inflammatory cascades.In the present study,Tp0751 recombinant protein from T.pallidum was found to induce the production of proinflammatory cytokines,including TNF-α,IL-1βand IL-6,in a THP-1 human monocyte cell line.The signal transduction pathways involved in the production of these cytokines were then further investigated.No inhibition of TNF-a,IL-1β,or IL-6 production was observed following treatment with the SAPK/JNK specific inhibitor SP600125 or with an ERK inhibitor PD98059.By contrast,anti-TLR2 mAb,anti-CD14 mAb,and the p38 inhibitor SB203580 significantly inhibited the production of all three cytokines.In addition,pyrrolidine dithiocarbamate (PDTC),a specific inhibitor of NF-κB,profoundly inhibited the production of these cytokines.Tp0751 treatment strongly activated NF-κB,as revealed by Western blotting.However,NF-κB translocation was significantly inhibited by treatment with PDTC.These results indicated that TLR2,CD14,MAPKs/p38,and NF-κB might be implicated in the inflammatory reaction caused by T.pallidum infection. | LIU ShuangQuan1,2,WANG ShiPing1,WU YiMou3,ZHAO FeiJun3,ZENG TieBing3,ZHANG YueJun3,ZHANG QiuGui2 & GAO DongMei1 1 Department of Parasitology,Xiangya School of Medicine,Central South University,Changsha 410078 ,China 2 The First Affiliated Hospital,University of South China,Hengyan 421001,China 3 Department of Microbiology and Immunology,University of South China,Hengyan 421001,China | 2010 | Science China(Life Sciences)2010，53，2： | 28 |
| 8 | Screening and breeding of high taxol producing fungi by genome shuffling显示文摘To apply the fundamental principles of genome shuffling in breeding of taxol-producing fungi, Nodulisporium sylviform was used as starting strain in this work. The procedures of protoplast fusion and genome shuffling were studied. Three hereditarily stable strains with high taxol production were obtained by four cycles of genome shuffling. The qualitative and quantitative analysis of taxol produced was confirmed using thin-layer chromatography (TLC), high performance liquid chromatography (HPLC) and LC-MS. A high taxol producing fungus, Nodulisporium sylviform F4-26, was obtained, which produced 516.37 μg/L taxol. This value is 64.41% higher than that of the starting strain NCEU-1 and 31.52%―44.72% higher than that of the parent strains. | ZHAO Kai, PING WenXiang, ZHANG LiNa, LIU Jun, LIN Yan, JIN Tao & ZHOU DongPo Laboratory of Microbiology, College of Life Science, Heilongjiang University, Harbin 150080, China | 2008 | Science China(Life Sciences)2008，51，3： | 25 |
| 9 | Deletion of Helicobacter pylori vacuolating cytotoxin gene by introduction of directed mutagenesis显示文摘AIM: To construct a vacA-knockout Helicobacter pylori mutant strain, whose only difference from the wild strain is its disrupted vacA gene.METHODS AND RESULTS: A clone containing kanamycin resistance gene used for homologous recombination was constructed in a directional cloning procedure into pBluescript ⅡSK, and then transformed into vacA+ H pylori by electroporation.Colonies growing on the selective media containing kanamycin were harvested for chromosomal DNA extraction,and the allelic exchange was determined by polymerase chain reactions and sequencing. Loss of vacuolating activity of the vaci-knockout strain was confirmed by examining the gastric cells co-cultured with cell-free supernatants from H pylori wild strain or the mutant.CONCLUSION: We constructed a vacA-knockout strain of H pylori through direct mutagenesis, which creates an important precondition for the future research on virulence comparison with gene expression analysis. | Jian-Ping Yuan,Tao Li,Xiao-Dong Shi,Bao-Yu Hu,Gui-Zhen Yang,Shan-Qing Tong,Xiao-Kui Guo,Department of Microbiology and Parasitology,Shanghai Second Medical University,Shanghai 200025,China. | 2003 | World Journal of Gastroenterology2003，9，10： | 20 |
| 10 | TRAF-mediated regulation of immune and inflammatory responses显示文摘The tumor necrosis factor (TNF) receptor-associated factor (TRAF) family consists of six mammalian members,and is shown to participate in signal transduction of a large number of receptor families including TNF receptor family (TNFR) and Toll-like receptors-interleukin-1 receptors (TLR-IL-1R) family.Upon receptor activation,TRAFs are directly or indirectly recruited to the intracellular domains of these receptors.They subsequently engage other signaling proteins to activate inhibitor of κB kinase (IKK) complex,TRAF family member-associated NF-κB activator (TANK)-binding kinase 1 (TBK1) and inducible I κB kinase (IKK-i) (also known as IKKε),ultimately leading to activation of transcription factors such as NF-κB and interferon-regulatory factor (IRF) to induce immune and inflammatory responses. | WANG YaYa1,ZHANG Peng2,LIU YingFang2 & CHENG GenHong1 1 Department of Microbiology,Immunology & Molecular Genetics,University of California Los Angeles,609 Charles E.Young Drive East,Los Angeles,California 90095,USA 2 National Laboratory of Biomacromolecules,Institute of Biophysics,Chinese Academy of Sciences,Beijing 100101,China | 2010 | Science China(Life Sciences)2010，53，2： | 19 |
| 11 | Antibiotic Resistance of Probiotic Strains of Lactic Acid Bacteria Isolated from Marketed Foods and Drugs显示文摘Objective To identify the antimicrobial resistance of commercial lactic acid bacteria present in microbial foods and drug additives by analyzing their isolated strains used for fermentation and probiotics. Methods Antimicrobial susceptibility of 41 screened isolates was tested with disc diffusion and E-test methods after species-level identification. Resistant strains were selected and examined for the presence of resistance genes by PCR. Results Distribution of resistance was found in different species. All isolates were susceptible to chloramphenicol, tetracycline, ampicillin, amoxicillin/clavulanic acid, cephalothin, and imipenem. In addition, isolates resistant to vancomycin, rifampicin, streptomycin, bacitracin, and erythromycin were detected, although the incidence of resistance to these antibiotics was relatively low. In contrast, most strains were resistant to ciprofloxacin, amikacin, trimethoprim/sulphamethoxazole, and gentamycin. The genes msrC, vanX, and dfrA were detected in strains of Enterococcus faecium, Lactobacillus plantarum, Streptococcus thermophilus, and Lactococcus lactis. Conclusion Antibiotic resistance is present in different species of probiotic strains, which poses a threat to food safety. Evaluation of the safety of lactic acid bacteria for human consumption should be guided by established criteria, guidelines and regulations. | CHANG LIU, ZHUO-YANG ZHANG, KE DONG, JIAN-PING YUAN, AND XIAO-KUI GUO P1 P Department of Medical Microbiology and Parasitology, Institutes of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China | 2009 | Biomedical and Environmental Sciences2009，22，5： | 19 |
| 12 | Humoral and cellular immunogenecity of DNA vaccine based on hepatitis B core gene in rhesus monkeys显示文摘INTRODUCTIONHepatitis B virus (HBV) is the most commonetiologic agent for infectious liver diseases. It isestimated that there are more than 250 millionchronic HBV carriersin the world today and thereis a significant association among persistentinfection, liver cirrhosis and hepatocellularcarcinoma[1-3]. | Zu Hu Huang1 Hui Zhuang2 Shan Lu3 Ren Hua Guo1 Guo Min Xu2 Jie Cai1 Wan Fu Zhu2 1Department of Infectious Diseases. The First Affiliated Hospital of Nanjing Medical University, Nenjing 210029, Jiangsu Province. China2Faculty of Microbiology, Beijing University, Beijing 100000, China3University of Massachusetts Medical Center | 2001 | World Journal of Gastroenterology2001，7，1： | 19 |
| 13 | Virulence and potential pathogenicity of coccoid Helicobacter pylori induced by antibiotics显示文摘AIM To explore the virulence and the potential pathogenicity of coccoid Helicobacter pylori (H. pylori) transformed from spiral form by exposure to antibiotic.METHODS Three strains of H. pylori, isolated from gastric biopsy specimens of confirmed peptic ulcer, were converted from spiral into coccoid from by exposure to metronidazole.Both spiral and coccoid form of H. pylori were tested for the urease activity, the adherence to Hep-2 cells and the vacuolating cytotoxicity to Hela cells, and the differences of the protein were analysed by SDS-PAGE and Western blot,The mutation of the genes including ureA, ureB,hpaA; vacA and cagA, related with virulence,was detected by means of PCR and PCR-SSCP.RESULTS In the coccoid H. pylori, the urease activity, the adherence to Hep-2 cells and the vacuolating cytotoxicity to Hela cells alldecreased. In strain F44, the rate and index of adherence reduced from 70.0% ± 5.3% to 33% ±5.1% and from 2.6 ±0.4 to 0.96 ±0.3 (P<0.01),respectively. The invasion of coccoid H. pylori into Hep-2 cell could be seen under electronmicroscope. SDS-PAGE showed that the content of the protein with the molecular weight over Mr74 000 decreased, and the hybriditional signal in band Mr 125 000 weakened, while the band Mr 110000 and Mr63000 strengthened in coccoid H. pylori as shown in Western blot. The results of PCR were all positive, and PCR-SSCP indicated that there may exist the point mutation in gene hpaA or vacA.CONCLUSION The virulence and the proteins with molecular weight over Mr74 000 in coccoid H. pylori decrease, but no deletion exists in amplification fragments from ureA, ureB, hpaA,vacA and cagA genes, suggesting that coccoid H. pylori may have potential pathogenicity. | Fei Fei She1 Dong Hui Su1 Jian Yin Lin2 Lin Ying Zhou3 1Department of Microbiology, Fujian Medical University. Fuzhou 350004, Fujian Province, China2Department of Molecular Medicine, Fujian Medical University, Fuzhou 350004, Fujian Province, China 3Laboratory of Electron Microscope, Fujian Medical University, Fuzhou 350004. Fujian Province. ChinaFei Fei She. graduated from Fujian Medical University as a postgraduate in 1991, now associate professor of microbiology and immunology, specialized in molecular biology of pathogen, having 15 papers published. | 2001 | World Journal of Gastroenterology2001，7，2： | 18 |
| 14 | Construction of hpaA gene from a clinical isolate of Helicobacter pyloriand identification of fusion protein显示文摘AIM: To clone hpaA gene from a clinical strain of Helicobacter pylori and to construct the expression vector of the gene and to identify immunity of the fusion protein.METHODS: The hpaA gene from a clinical isolate Y06 of H.pylori was amplified by high fidelity PCR. The nucleotide sequence of the target DNA amplification fragment was sequenced after T-A cloning. The recombinant expression vector inserted with hpaA gene was constructed. The expression of HpaA fusion protein in E. coli BL21DE3 induced by IPTG at different dosages was examined by SDS-PAGE.Western blot with commercial antibody against whole cell of H. pylorias well as immunodiffusion assay with selfprepared rabbit antiserum against HpaA fusion protein were applied to determine immunity of the fusion protein. ELISA was used to detect the antibody against HpaA in sera of 125 patients infected with H. pylori and to examine HpaA expression of 109 clinical isolates of H. pylori.RESULTS: In comparison with the reported corresponding sequences, the homologies of nucleotide and putative amino acid sequences of the cloned hpaA gene were from 94.25-97.32 % and 95.38-98.46 %, respectively. The output of HpaA fusion protein in its expression system of pET32ahpaA-BL21DE3 was approximately 40 % of the total bacterial proteins. HpaA fusion protein was able to combine with the commercial antibody against whole cell of H. pyloriand to induce rabbit producing specific antiserum with 1:4immunodiffusion titer after the animal was immunized with the fusion protein. 81.6 % of the serum samples from 125patients infected with H.pylori(102/125) were positive for HpaA antibody and all of the tested isolates of H.pylori(109/109) were detectable for HpaA.CONCLUSION: A prokaryotic expression system with high efficiency of H.pylorihpaA gene was successfully established.The HpaA expressing fusion protein showed satisfactory immunoreactivity and antigenicity. High frequencies of HpaA expression in different H. pyloriclinicalstrains and specific antibody production in H. pyloriinfected patients indicate that HpaA is an excellent and ideal antigen for developing H. pylori vaccine. | Ya-Fei Mao Jie Yan Li-Wei Li Shu-Ping Li Department of Medical Microbiology and Parasitology,College of Medical Sciences,Zhejiang University,Hangzhou 310031,Zhejiang Province,China | 2003 | World Journal of Gastroenterology2003，9，7： | 17 |
| 15 | Immunization with chlamydial plasmid protein pORF5 DNA vaccine induces protective immunity against genital chlamydial infection in mice显示文摘To validate the immune protective efficacy of pORF5 DNA vaccine and to analyze potential mechanisms related to this protection. In this study, pORF5 DNA vaccine was constructed and evaluated for its protective immunity in a mouse model of genital chlamydial infection. Groups of BALB/c mice were immunized intranasally with pORF5 DNA vaccine. Humoral and cell mediated immune responses were evaluated. The clearance ability of chlamydial challenge from the genital tract and the chlamy- dia-induced upper genital tract gross pathology and histopathological characterization were also de- tected. The results showed that the total and the IgG2a anti-pORF5 antibody levels in serum were sig- nificantly elevated after pcDNA3.1-pORF5 vaccination, as were the total antibody and IgA levels in vaginal fluids. pcDNA3.1-pORF5 induced a significantly high level of Th1 response as measured by robust gamma interferon (IFN-γ). Minimal IL-4 was produced by immune T cells in response to the re-stimulation with pORF5 protein or the inactive elementary body in vitro. pcDNA3.1-pORF5-vacci- nated mice displayed significantly reduced bacterial shedding upon a chlamydial challenge and an accelerated resolution of infection. 100% of pcDNA3.1-pORF5 vaccinated mice successfully resolved the infection by day 24. pcDNA3.1-pORF5-immunized mice also exhibited protection against patho- logical consequences of chlamydial infection. The stimulated index was significantly higher than that of mice immunized with pcDNA3.1 and PBS (P<0.05). Together, these results demonstrated that immu- nization with pORF5 DNA vaccine is a promising approach for eliciting a protective immunity against a genital chlamydial challenge. | LI ZhongYu1,2, WANG ShiPing1, Wu YiMou2, ZHONG GuangMing3 & CHEN Ding3 1 Xiangya School of Medicine, Central South University, Changsha 410078, China 2 Department of Microbiology and Immunology, University of South China, Hengyang 421001, China 3 University of Texas Health Science Center at San Antonio, TX 78229, USA | 2008 | Science China(Life Sciences)2008，51，11： | 16 |
| 16 | Bacteriological and electron microscopic examination of primary intrahepatic stones显示文摘BACKGROUND: Primary intrahepatic cholelithiasis is usually combined with biliary tract infection. This research was undertaken to investigate the relationship between intrahepatic stones and biliary tract infection. METHODS: Thirty-five bile samples and 30 stones specimens were cultured for bacteria and 12 stones specimens were examined with a scan electron microscope (SEM) or a transmission electron microscope (TEM). RESULT: 94.2% bile samples and 96.7% stones specimens were positive in bacteria culture. Bacteria were found in stones under SEM and TEM. CONCLUSION: Bacteria in stones are associated with the infection of the biliary tract. | Shuo-Dong Wu, Hong Yu and Ji-Mei Sun Department of General Surgery Department of Microbiology Center Second Affiliated Hos- pital China Medical University, Shenyang 110004, China | 2006 | Hepatobiliary & Pancreatic Diseases International2006，5，2： | 16 |
| 17 | Characterizing and estimating rice brown spot disease severity using stepwise regression,principal component regression and partial least-square regression显示文摘Detecting plant health conditions plays a key role in farm pest management and crop protection. In this study, measurement of hyperspectral leaf reflectance in rice crop (Oryzasativa L.) was conducted on groups of healthy and infected leaves by the fungus Bipolaris oryzae (Helminthosporium oryzae Breda. de Hann) through the wavelength range from 350 to 2 500 nm. The percentage of leaf surface lesions was estimated and defined as the disease severity. Statistical methods like multiple stepwise regression, principal component analysis and partial least-square regression were utilized to calculate and estimate the disease severity of rice brown spot at the leaf level. Our results revealed that multiple stepwise linear regressions could efficiently estimate disease severity with three wavebands in seven steps. The root mean square errors (RMSEs) for training (n=210) and testing (n=53) dataset were 6.5% and 5.8%, respectively. Principal component analysis showed that the first principal component could explain approximately 80% of the variance of the original hyperspectral reflectance. The regression model with the first two principal components predicted a disease severity with RMSEs of 16.3% and 13.9% for the training and testing dataset, respec-tively. Partial least-square regression with seven extracted factors could most effectively predict disease severity compared with other statistical methods with RMSEs of 4.1% and 2.0% for the training and testing dataset, respectively. Our research demon-strates that it is feasible to estimate the disease severity of rice brown spot using hyperspectral reflectance data at the leaf level. | LIU Zhan-yu1, HUANG Jing-feng1, SHI Jing-jing1, TAO Rong-xiang2, ZHOU Wan3, ZHANG Li-li3 (1Institute of Agriculture Remote Sensing and Information System Application, Zhejiang University, Hangzhou 310029, China) (2Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China) (3Plant Inspection Station of Hangzhou City, Hangzhou 310020, China) | 2007 | Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)2007，8，10： | 13 |
| 18 | Isolation and Identification of a Specific cDNA Mapping to the Bam HI-I2 and -LFragments within the Inverted Repeats ofUnique Long Re-gion (IRL) in the Genom e ofMarek′s Disease Herpesvirus (MDV) Oncogenic Strain Beijing-1显示文摘Objective\ To understand the transcription of BamHI L DNA fragment from genome of strong virulent GA strain of Marek′s disease herpesvirus (MDV) in lymphoblastoid tumor tissue induced by oncogenic strain Beijing 1 (a specific local strain in China) of MDV. Methods\ Two oligonucleotide primers were synthesized according to the reported sequence of \%meq\% gene an ideal oncogenic candidate and our previously determined sequence of BamHI L fragment of Marek′s disease herpesvirus (MDV), respectively. Reverse transcriptase PCR(RT PCR) assay was performed by using these primers and the mRNA as a template which was isolated from visceral lymphoblastoid tumors obtained from chickens artificially infected with strain Beijing 1 of oncogenic MDV. Southern blot molecular hybridization was further carried out to detect the product of RT PCR with digoxigenin labeled nucleotide probe from BamHI I2 and L fragment in the gene library of MDV strain GA, respectively. Results\ Two probes could simultaneously hybridize this cDNA amplified by RT PCR with a length of about 730 bp. Conclusion\ It is suggested that \%meq\% transcription could extend from the right hand end of BamHI I2 to the adjacent BamHI L, and the BamHI L region was likely to be transcribed in MDV induced lymphoblastoid tumors. | Lu Chun, Wu Jianping, Zhang Xunhai, et al. Lu Chun, Wu Jianping, Zhang Xunhai, et al. Department of Microbiology and Immunology, Nanjing Medical University, Nanjing\ 210029 Acta Universitatis Medicinalis Nanjing, 19(6):447 | 1999 | The Journal of Biomedical Research1999，23，2： | 13 |
| 19 | Effect of Protein Kinase C on Proliferation and Apoptosis of T Lymphocytes in Idiopathic Thrombocytopenic Purpura Children显示文摘It is well-documented that T lymphocyte proliferation and apoptosis are abnormal in idiopathic thrombocytopenic purpura (ITP) children. However, the underlying regulation mechanisms especially in terms of signal transduction remain unknown. In this paper, we reported the changes of protein kinase C (PKC) activity in peripheral blood T lymphocytes and the effect of PKC on T lymphocyte proliferation and apoptosis. We demonstrated that in ITP children, the activator (PMA) and inhibitor (H-7) of PKC affected on T lymphocyte proliferation and apoptosis dramatically, but they altered little in healthy children. PKC activity was significantly enhanced in ITP children together with an increased expression of FasL on CD3+ T, CD4+ T and CD8+T cells, resulting in a positive correlation between PKC activity and the expression of FasL on T cells. While the PKC activity and the platelet count were negatively correlated. Taken together, our findings suggest that the PKC activation may enhance T lymphocytes activity, suppress T cell apoptosis and be involve in thrombocytes damage as a mechanism related to immune pathogenesis of ITP. | Changlin Wu~(1,2) Fang Liu~1 Xuemin Zhou~1 Zhengwei Cheng~1 Xiaomeng Yang~1 Hong Xiao~1 Qun Chen~1 Kangrong Cai~1 ~1Department of Microbiology and Immunology,Guangdong Medical College,Zhanjiang,Guangdong,524023,China ~2Corresponding to:Dr.Changlin Wu,Department of Microbiology and Immunology,Guangdong Medical College,2 Wen-Ming East Road,Zhanjiang,Guangdong,524023,China. | 2005 | Cellular & Molecular Immunology2005，2，3： | 12 |
| 20 | Stability of randomly amplified polymorphic DNA fingerprinting in genotyping clinical isolates of Helicobacter pylori显示文摘AIM: Hpylorigenomes are highly diversified. This project was designed to genotype Hpyloriisolates by the polymerase chain reaction (PCR)-based randomly amplified polymorphic DNA (RAPD) fingerprinting technique and to verify its stability by Southern blotting and DNA sequencing.METHODS: Clinical isolates of Hpyloriwere cultured from gastric antra and cardia of 73 individuals, and genomic DNA was prepared for each isolate. RAPD was carried out under optimized conditions. 23S rDNA was regarded as an internal control, and a 361 bp rDNA fragment (RDF) was used as a probe to screen the RAPD products by Southern blotting.Ten RDFs from different clinical isolates and the flanking regions (both upstream and downstream) of four RDFs were amplified and sequenced.RESULTS: Hpyloriisolates from different individuals had different RAPD profiles, but the profiles for isolates cultured from different gastric sites of a given individual were identical in all but one case. Isolates from 27 individuals were RDF positive by Southern blotting. Sequences of the RDFs and their flanking regions were almost the same between the RDF positive and negative isolates as determined by Southern blotting. There was no binding site for random PCR primer inside the sequences.CONCLUSION: RAPD is very useful in genotyping H pylori grossly on a large scale. However, it seems unstable in amplification of low yield fragments, especially those that do not appear as visible bands on the agarose gel stained with EB, since the palmer is partially matched to the template. | Feng-Chan Han Han-Chong Ng Bow Ho Department of Microbiology,Faculty of Medicine,National University of Singapore,5 Science Drive 2,Singapore 117595,Republic of Singapore | 2003 | World Journal of Gastroenterology2003，9，9： | 9 |
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