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Identification and classification of all potential hemolysin encoding genes and their products from Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai

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Acta Pharmacologica Sinica 2005年第4期26卷 453-461页

作者： Yi-xuan ZHANG~(2,3,4,5) Yan GENG(2,3,4,5) Bo BI~(2,4) Jian-yong HE~4 Chun-fu WU~4 Xiao-kui GUO~3 Guo-ping ZHAO~(2,4,6,7) 2 Research center of Biotechnology,shanghai Institutes for Biological Sciences,chinese Academy of Sciences,shanghai 200233,china 3 Department of Microbiology and Parasitology,shanghai Second Medical University,shanghai 200025,china 4 The School of Pharmaceutical Engineering,Shenyang Pharmaceutical University,Shenyang 110016,china 6 chinese national human genome center at shanghai,ZhangJiang High Tech Park,shanghai 201203,china Research center of Biotechnology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China Department of Microbiology and Parasitology Shanghai Second Medical University Shanghai China The School of Pharmaceutical Engineering Shenyang Pharmaceutical University Shenyang China chinese national human genome center at shanghai ZhangJiang High Tech Park Shanghai China

Aim:To identify and classify all potential hemolysin candidates of Leptospirainterrogans serogroup Icterohaemorrhagiae serovar ***:All of thepotential hemolysin encoding genes were characterized in *** geneswere cloned and expressed in Escherichia *** hemolytic activities of theexpressed proteins were assayed observing the hemolysis on sheep blood *** activities of the hemolysin candidates were measuredby thin-layer chromatography(TLC)and HPLC for *** and secretion of the hemolysins in L interrogans were studied byreverse transcription polymerase chain reaction,Western blot,and enzyme-linkedimmunosorbent *** and Conclusion:The hemolytic activities ofhemolysin candidates(LA0327,LA0378,LA1027,LA1029,LAI650,LA3050,LA3937,LA4004)from L interrogans strain Lai were *** were furtherdivided into two groups,sphingomyelinase hemolysins and non-sphingomyelinasehemolysins,based on their ability to hydrolyze *** of thesehemolysins were actually expressed in living L interrogans and some of themwere secreted into the *** study establishes an essential findcomplete basis for further studying the contribution of hemolysins to the patho-genesis of L interrogans.

关键词： hemolysin Leptospira interrogans sphingo myelinase

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Progress in the detection of human genome structural variations

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Science china(Life Sciences) 2009年第6期52卷 560-567页

作者： WU XueMei1,3 & XIAO HuaSheng1,2 1 center of Functional genomics,Key laboratory of System Biology,shanghai Institutes for Biological Sciences,chinese Academy of Sciences,shanghai 200031,china 2 national Engineering center for Biochip at shanghai,shanghai 201203,china 3 Graduate School of the chinese Academy of Sciences,shanghai 200031,china center of Functional genomics Key Laboratory of System Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China Graduate School of the chinese Academy of Sciences Shanghai China national Engineering center for Biochip at shanghai Shanghai China

The emerging of high-throughput and high-resolution genomic technologies led to the detection of submicroscopic variants ranging from 1 kb to 3 Mb in the human *** variants include copy number variations(CNVs),inversions,insertions,deletions and other complex rearrangements of DNA *** paper briefly reviews the commonly used technologies to discover both genomic structural variants and their potential ***,we highlight the array-based,PCR-based and sequencing-based assays,including array-based comparative genomic hybridization(aCGH),representational oligonucleotide microarray analysis(ROMA),multiplex amplifiable probe hybridization(MAPH),multiplex ligation-dependent probe amplification(MLPA),paired-end mapping(PEM),and next-generation DNA sequencing ***,we discuss the limitations and challenges of current assays and give advices on how to make the database of genomic variations more reliable.

关键词： structural variation cytogenetic microarray PCR paired end mapping(PEM) next-generation sequencing

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Bacterial Type Ⅰ Glutamine Synthetase of the Rifamycin SV Producing Actinomycete,Amycolatopsis mediterranei U32,is the Only Enzyme Responsible for Glutamine Synthesis under Physiological Conditions

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Acta Biochimica et Biophysica Sinica 2006年第12期 821-830页

作者： Wen-Tao PENG~(1&) Jin WANG~1 Ting WU~2 Jian-Qiang HUANG~1 Jui-Shen CHIAO~1 Guo-Ping ZHAO~(1,3,4) 1 laboratory of Molecular Microbiology,Institute of Plant Physiology and Ecology,shanghai Institutes for Biological Sciences,chinese Academy of Sciences,shanghai 200032,china 2 Department of Medicinal Chemistry,School of Pharmacy,Fudan University,shanghai 200032,china 3 State Key laboratory of Genetic Engineering/Department of Microbiology,School of Life Science,Fudan University,shanghai 200433,china 4 laboratory of health and disease genomics,chinese national Haman genome center at shanghai,shanghai 201203,china

The structural gene for glutamine synthetase,glnA,from Amycolatopsis mediterranei U32 wascloned via screening a genomic library using the analog gene from Streptomyces *** clone wasfunctionally verified by complementing for glutamine requirement of an Escherichia coli glnA null mutantunder the control of a lac *** analysis showed an open reading frame encoding a protein of466 amino acid *** deduced amino acid sequence bears significant homologies to other bacterialtype Ⅰ glutamine synthetases,specifically,71% and 72% identical to the enzymes of *** and Myco-bacterium tuberculosis,*** of this glnA gene in *** U32 led to glutamineauxotrophy with no detectable glutamine synthetase activity in *** contrast,the cloned glnA+gene cancomplement for both phenotypes in *** thus suggested that in *** U32,the glnA geneencoding glutamine synthetase is uniquely responsible for in vivo glutamine synthesis under our laboratorydefined physiological conditions.

关键词： glnA glutamine synthetase Amycolatopsis mediterranei rifamycin SV

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Identification and Analysis of Genes Present in Leptospira interrogans serovar lai but Absent in L. biflexa serovar monvalerio

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Acta Biochimica et Biophysica Sinica 2004年第12期36卷 832-839页

作者： Ping HE, Xiang-Yan ZHANG, Xiao-Kui GUO*, Bao-Yu HU, Xiao-Tian HUANG, Yang YANG, and Guo-Ping ZHAO1,2* Department of Microbiology and Parasitology, shanghai Second Medical University, shanghai 200025, china 1 Research center of Biotechnology, Institutes for Biological Sciences, chinese Academy of Sciences, shanghai 200031, china 2chinese national human genome center at shanghai, Zhangjiang High Tech Park, shanghai 201203, china Department of Microbiology and Parasitology Shanghai Second Medical UniversityShanghai 200025 China Research center of Biotechnology Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200031 China chinese national human genome center at shanghaiZhangjiang High Tech Park Shanghai 201203 China

Genes present in virulent bacterial strains but absent in avirulent close relatives can be of great biologic and clinical interest. This project aimed to identify strain specific DNA sequences of Leptospira interrogans serovar lai, which is absent in the saprophytic L. biflexa serovar monvalerio, via suppression subtractive hybridization with the former as the tester while the latter as the driver. The mixture of PCR amplified DNA fragments from two subtractive hybridization experiments were cloned into pMD18-T vector and the positive clones were identified by dot blotting against the chromosome DNA of the two strains individually. After DNA sequencing and analysis, the distribution of these genomic fragment sequences in a panel of pathogenic and nonpathogenic leptospires was investigated employing dot blot analysis. Among the 188 positive clones randomly chosen, 24 contained the tester strain specific genomic regions, of which, 5 were non-coding fragments while the others contained 23 distinct protein coding sequences. Besides 9 genes encoding functional proteins, 12 genes encode unknown proteins and the rest two genes encode proteins with recognizable domain structures, one for a putative leucine-rich repeats (LRR) family protein while the other as an outer-membrane protein. Our experiment results indicated that suppression subtractive hybridiza- tion is effective for screening specific DNA sequences between two leptospiral strains, and some of these sequences might be responsible for virulence determination. Further analysis of these DNA sequences will provide important information on the pathogenesis of Leptospira.

关键词： Leptospira interrogans serovar lai suppression subtractive hybridization (SSH) specific genes

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中国百日咳诊疗与预防指南(2024版)

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中华医学杂志 2024年第15期104卷 1258-1279页

作者：中华医学会感染病学分会儿科感染学组国家卫生健康委能力建设和继续教育儿科专委会感染组中国临床实践指南联盟方法学专委会国家儿童医学中心(上海) 国家传染病医学中心曾玫邵祝军张文宏夏君蔡洁皓陈明亮冯天行姚开虎不详复旦大学附属儿科医院感染传染科国家儿童医学中心(上海)上海200032 中国疾病预防控制中心传染病预防控制所北京102206 复旦大学附属华山医院感染科国家传染病医学中心上海200040 宁波诺丁汉大学GRADE中心宁波315100 复旦大学附属儿科医院感染传染科复旦大学附属闵行医院检验科上海临床研究中心儿科首都医科大学附属北京儿童医院北京市儿科研究所感染与微生物研究室国家儿童医学中心(北京)

百日咳再现是全球性的公共卫生问题,2018—2022年我国国内百日咳报告发病率接近20世纪80年代末的报告水平。实际上,我国的百日咳发病率被严重低估,主要因为国内尚未全面建立百日咳主动监测工作,不典型百日咳病例在临床上容易被漏诊,许... 详细信息

百日咳再现是全球性的公共卫生问题,2018—2022年我国国内百日咳报告发病率接近20世纪80年代末的报告水平。实际上,我国的百日咳发病率被严重低估,主要因为国内尚未全面建立百日咳主动监测工作,不典型百日咳病例在临床上容易被漏诊,许多医疗机构未开展百日咳的实验室诊断;同时,我国还面临百日咳鲍特菌对一线抗菌药物大环内酯类高度耐药的临床问题。为了更好地指导和规范我国百日咳病例的临床诊疗、监测和防控工作,由感染病学、流行病学、免疫规划及指南方法学专家组成指南制订小组,从实践角度提出有关百日咳诊断、治疗、预防,尤其是疫苗免疫策略的12个临床问题,经过研究问题构建、证据检索和综合、证据评价以及证据到推荐意见讨论,制订推荐意见和实施建议,为从事百日咳诊疗和防控的临床医师、微生物实验室专业人员、医院感染控制专业人员以及公共卫生领域从事传染病防控和免疫规划的专业人员提供参考。

关键词：百日咳诊断治疗预防疫苗接种

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Characterization of a novel toxin-antitoxin module, VapBC, encoded by Leptospira interrogans chromosome

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Cell Research 2004年第3期14卷 208-216页

作者： Yi Xuan ZHANG, Xiao Kui GUO, Chuan WU, Bo BI, Shuang Xi REN, Chun Fu WU, Guo Ping ZHAO Pharmaceutical Department, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenhe District, Shenyang 110016, china. Research center of Biotechnology, shanghai Institutes for Biological Sciences, chinese Academy of Sciences, 500 Caobao Road, shanghai 200233, china. Department of Microbiology and Parasitology, shanghai Second Medical University, 280 Chongqingnan Road, shanghai 200025, china. chinese national human genome center at shanghai (CHGCS), 250 Bibo Road, ZhangJiang High Tech Park, shanghai, 201203, china. Pharmaceutical Department Shenyang Pharmaceutical University 103 Wenhua Road Shenhe District Shenyang China Research center of Biotechnology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China Department of Microbiology and Parasitology Shanghai Second Medical University Shanghai China chinese national human genome center at shanghai (CHGCS) 250 Bibo Road ZhangJiang High Tech Park Shanghai China

Comparative genomic analysis of the coding sequences （CDSs） of Leptospira interrogans revealed a pair of closely linked genes homologous to the vapBC loci of many other bacteria with respect to both deduced amino acid sequences and operon organizations. Expression of single vapC gene in Escherichia coli resulted in inhibition of bacterial growth, whereas co-expression of vapBC restored the growth effectively. This phenotype is typical for three other characterized toxin-antitoxin systems of bacteria, i.e., mazEF[1], relBE[2] and chpIK[3]. The VapC proteins of bacteria and a thermophilic archeae, Solfolobus tokodaii, form a structurally distinguished group of toxin different from the other known toxins of bacteria. Phylogenetic analysis of both toxins and antitoxins of all categories indicated that although toxins were evolved from divergent sources and may or may not follow their speciation paths （as indicated by their 16s RNA sequences）, co-evolution with their antitoxins was obvious.

关键词： Leptospria toxin-antitoxin system VapBC co-evolution speciation.

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Design,synthesis,antitumor evaluations and molecular modeling studies of novel 3,5-substituted indolin-2-one derivatives

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Acta Pharmacologica Sinica 2007年第1期28卷 140-152页

作者： Hai-hong LI~2 Xiu-hua ZHANG~3 Jin-zhi TAN~2 Li-li CHEN~2 Hong LIU~(2,5) Xiao-min LUO~(2,5) Xu SHEN~(2,4) Li-ping LIN~(3,5) Kai-xian CHEN~2 Jian DING~3 Hua-liang JIANG~(2,4)2 center for Drug Discovery and Design,State Key laboratory of Drug Research,shanghai Institute of Materia Medica,shanghai Institutes for Biological Sciences,and Graduate School,chinese Academy of Sciences,shanghai 201203,china 3 Division of Anti-tumor Pharmacology,shanghai Institute of Materia Medica,shanghai Institutes for Biological Sciences,chinese Academy of Sciences,shanghai 201203,china 4 School of Pharmacy,East china University of Science and Technology,shanghai 200237,china center for Drug Discovery and Design State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences and Graduate School Chinese Academy of Sciences Shanghai China Division of Anti-tumor Pharmacology 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

Aim:To design and synthesize a novel class of antitumor agents,featuring the 3,5-substituted indolin-2-one ***:Based on enzyme binding fea-tures of（Z）-SU5402,introducing a β-pyrrole group at the 3-position of the indolin-2-one core,a series of novel 3,5-substituted indolin-2-ones were designed *** human carcinoma cell lines of A-43l,A-549,MDA-MB-468,and Autosomal Dominant Polycystic Kidney disease were chosen for the cellproliferation ***:Twenty new compounds（1a-t）with E configurationhave been designed,synthesized and *** structural features weredetermined by nuclear magnetic resonance（NMR）spectra,low-and high-resolu-tion mass spectra,and confirmed by X-ray *** the enzymeassay showed a weak inhibition effect against the epidermal growth factor receptor,vascular endothelial growth factor receptor,fibroblast growth factor receptor andplatelet-derived growth factor receptor tyrosine kinases,the cell-based antitumoractivity was *** 1 g and lh showed higher inhibitory activitytoward the A-549 and MDA-MB-468 cell lines with IC50of 0.065-9.4 μmol/***:This study provides a new template for further development ofpotent antitumor drugs.

关键词： protein-tyrosine kinase indolin-2-one antitumor drug screening assays drug design molecular models

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Construction of a small peptide library related to inhibitor OM99-2 and its structure-activity relationship to β-secretase

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Acta Pharmacologica Sinica 2006年第12期27卷 1586-1593页

作者： Bin HU Bing XIONG Bei-ying QIU Xin LI Hai-ping YU Kun XIAO Xin WANG Jia LI Jing-kang SHEN~2 State Key laboratory of Drug Research and national center for Drug Screening,shanghai Institute of Materia Medica,shanghai Institutes for Biological Sciences,chinese Academy of Sciences,shanghai 201203,china State Key laboratory of Drug Research and national center for Drug Screening Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

Aim:To develop probes for detecting the binding specificity between β-secretaseand substrate,and provide reliable biological activity data for further researchingencircling substrate-based ***:To prepare the inhibitors,thehydroxyethylene(HE)segment including P1and P1′ was synthesized after multi-step reactions;the combination of all segments was then completed through solidphase *** human β-secretase ectodomain(amino acid resi-dues 1-460)was expressed as a secreted protein with a C-terminal His tag in insectcells using baculovirus infection,and all compounds were evaluated in thisβ-secretase enzyme *** order to understand the interaction in detail,the theoretical methods,namely molecular dynamics(MD)simulation and mo-lecular mechanics-generalized-born surface area(MM-GBSA)analysis,were per-formed on the complex of β-secretase and OM99-2 to obtain the geometrical andenergetical ***:We designed and constructed a positional scan-ning combinatorial library including 16 compounds;all members of the librarywere synthesized based on HE dipeptide ***-activity relationshipstudies at the P4-P1and P1′-P4′ positions led to the discoveries of P and P’sidesbinding specificity and potent inhibitors 14,18,and *** binding free energyon the whole system and every residue were compared to the biological ***:The removal of P4′ yielded inhibitor 22(A3*B2)with highpotency;further truncation of P3′ gave inhibitor 18(A3*B1)with equal activity,implying that the right side of the inhibitors play a less important role and could beeasily simplified,while change on the P side may cause substantial results.

关键词： β-secretase hydroxyethylene OM99-2 Alzheimer’s disease structure-activity relationship

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Preliminary studies on differential defense responses induced during plant communication

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Cell Research 2005年第3期15卷 187-192页

作者： Jin Ying PENG1,3,Zhong Hai LI2,Hui XIANG1,3,Jian Hua HUANG1,3,Shi Hai JIA1,3,Xue Xia MIAO1,Yong Ping HUANG1* 1Institute of Plant Physiology and Ecology,shanghai Institute for Biological Sciences,Graduate School of the chinese Academy of Sciences,chinese Academy of Sciences,300 Fenglin Road,shanghai 200032,china 2chinese national human genome center at shanghai,250 Bibo Road,shanghai 201203,china 3Graduate School of the chinese Academy of Sciences,Beijing 100039,china Institute of Plant Physiology and Ecology Shanghai Institute for Biological Sciences Graduate School of the Chinese Academy of Sciences Chinese Academy of Sciences Shanghai China Graduate School of the chinese Academy of Sciences Beijing China chinese national human genome center at shanghai Shanghai China

We compared the expression patterns of three representative genes in undamaged tomato and tobacco plants in response to exposure to either tomato or tobacco fed on by Helicoverpa armigera (cotton bollworm). When tomato and tobacco, two species of one family, were incubated in the chambers with the tomato plants damaged by the cotton bollworm, the expression of the PR1, BGL2, and PAL genes was up-regulated in leaves of both plants. However, the levels of gene expression were significantly higher in the tomato than that in the tobacco. In addition, the activities of enzymes, peroxidase, polyphenol oxidase, and lipoxygenase were found to be higher in the tomato than those in the tobacco. Similar results were obtained when the damaged plants were replaced by the tobacco.

关键词： plant-plant communication tomato (Lycopersicon esculentum) tobacco (Nicotiana tabacum) cotton boll- worm (Helicoverpa armigera) inducible defenses.

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Association of DRD2 polymorphisms and chlorpromazine-induced extrapyramidal syndrome in chinese schizophrenic patients

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Acta Pharmacologica Sinica 2006年第8期27卷 966-970页

作者： Sheng-nan WU~(2,3,5,6) Rui GAO~(2,3,6) Qing-he XING~(2,3,6) Hua-fang LI~4 Yi-feng SHEN~4 Niu-fan GU~4 Guo-yin FENG~4 Lin HE~(2,3,7)2 Bio-X Life Science Research center,shanghai Jiaotong University,shanghai 200030,china 3 Institute of Nutritional Sciences,SIBS,chinese Academy of Sciences,shanghai 200031,china 4 shanghai Institute of Mental health,shanghai 200030,china 5 shanghai Institute of Hypertension,Ruijin Hospital,Medical School of shanghai Jiaotong University,shanghai 200025,china Bio-X Life Science Research center Shanghai Jiaotong University Shanghai China Institute of Nutritional Sciences SIBS Chinese Academy of Sciences Shanghai China shanghai Institute of Hypertension Ruijin Hospital Medical School of Shanghai Jiaotong University Shanghai China shanghai Institute of Mental health Shanghai China

Aim:Extrapyramidal syndrome (EPS) is most commonly affected by typical antip-sychotic drugs that have a high affinity with the D2 ***,manyresearch groups have reported on the positive relationship between the geneticvariations in the DRD2 gene and the therapeutic response in schizophrenia pa-tients as a result of the role of variations in the receptor in modulating *** this study,we evaluate the role DRD2 plays in chlorpromazine-induced EPS in schizophrenic ***:We identified seven SNP(singlenucleotide polymorphism) (-141Cins>del,TaqIB,TaqID,Ser311Cys,rs6275,rs6277and TaqIA) in the DRD2 gene in 146 schizophrenic inpatients (59 with EPS and 87without EPS according to the Simpson-Angus Scale) treated with chlorpromazineafter 8 *** alleles of all loci were determined by PCR (polymerase chainreaction).Results:Polymorphisms TaqlD,Ser311Cys and rs6277 were not poly-morphic in the population recruited in the present *** statistical signifi-cance was found in the allele distribution of-141Cins>del,TaqIB,rs6275 andTaqIA or in the estimated haplotypes (constituted by TaqIB,rs6275 and TaqIA) inlinkage disequilibrium between the two ***:Our results did notlend strong support to the view that the genetic variation of the DRD2 gene playsa major role in the individually variable adverse effect induced by chlorpromazine,at least in chinese patients with *** results confirmed a previousstudy on the relationship between DRD2 and EPS in Caucasians.

关键词： dopamine receptors basal ganglia disease polymorphism chlorpromazine schizophrenia

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