简介:目的:在pEGFP-C2真核表达载体中表达人NDRG2基因的截短部分并进行鉴定。方法:以含有人NDRG2基因的pGKBT7质粒为模板,通过PCR的方法扩增获得截短的人NDRG2基因,然后克隆入表达载体pEGFP-C2;以荧光显微镜观察和WesternBlotting方法对表达产物进行鉴定。结果:表达产物中在分子量67kD左右可见与目的蛋白分子量相符的条带,该条带可被Ndrg2单克隆抗体特异性识别。结论:构建了截短的人NDRG2基因真核表达载体,并在真核细胞中(HEK-293)获得成功表达。
简介:目的:探究胃炎康方5号联合西医治疗慢性萎缩性胃炎的临床疗效。方法:选取在我院接受治疗的慢性萎缩性胃炎患者94例,分为对照组和实验组,对照组患者采用常规的胃三联治疗方法,实验组患者在对照组患者的治疗方法之上再加上胃炎康方5号治疗,观察分析和对比两组患者的治疗效果以及Hp根除率(幽门螺旋杆菌根除率)。结果:对照组患者治疗的总有效率为46.81%,Hp根除率为82.98%,实验组患者治疗的总有效率为80.85%,根除率为93.62%,两组结果相比较,差异显著,有统计学意义(P<0.05)。结论:胃炎康方5号联合西药治疗慢性萎缩性胃炎的临床疗效显著,值得临床推广。
简介:ErbB2,amemberofthereceptortyrosinekinasefamily,isfrequentlyover-expressedinbreastcancer.ProteolysisoftheextracellulardomainofErbB2resultsinconstitutiveactivationofErbB2kinase.RecentstudyreportedthatErbB2isfoundinthenucleus.Here,weshowedthatErbB2isimportedintothenucleusthroughanuclearlocalizationsignal(NLS)-mediatedmechanism.TheNLSsequenceKRRQQKIRKYTMRR(aa655-668)containsthreeclustersofbasicaminoacidsanditissufficienttotargetGFPintothenucleus.However,mutationinanybasicaminoacidclusterofthisNLSsequencesignificantlyaffectsitsnuclearlocalization.Furthermore,itwasfoundthatthisNLSisessentialforthenuclearlocalizationofErbB2sincetheintracellulardomainofErb2lackingNLScompletelyabrogatesitsnucleartranslocation.Takentogether,ourstudyidentifiedanovelnuclearlocalizationsignalandrevealsanovelmechanismunderlyingErbB2nucleartraffickingandlocalization.
简介:以土生鳞伞(PholiotaterrestrisOverh.)子实体为生物吸附剂吸附水溶液中的Cd2+,分析吸附剂用量、初始pH值、初始重金属浓度、反应时间这4个因素对吸附的影响,并采用Langmuir和Freundlich等温吸附模型及准一级、准二级动力学模型拟合土生鳞伞的生物吸附特性。结果表明:水溶液中Cd2+的去除率随吸附剂用量和反应时间的增加而增大,当Cd2+的浓度为10g/L时,土生鳞伞子实体吸附剂用量为2g/L,反应2h即可达到平衡吸附,且pH=6时吸附量最大。土生鳞伞子实体对Cd2+的吸附更符合Langmuir平衡模型,相关系数r2>0.99,土生鳞伞子实体对Cd2+的最大吸附量为8.67mg/g。准二级动力学方程比准一级动力学方程能更好地描述土生鳞伞子实体对Cd2+的吸附。
简介:包括谷物尺寸和圆锥花序形态学,圆锥花序的建筑学直接决定谷物产量。圆锥花序直立,为在中国的北部分完成理想的植物建筑学被选择,引起了米饭breeders的增加的注意。这里,稠密、直立的圆锥花序2(dep2)异种,显示出稠密、直立的圆锥花序显型,被识别。没有任何已知的功能的领域,DEP2编码植物特定的蛋白质。表示介绍DEP2表明它高度在年轻纸巾被表示,与在年轻圆锥花序的大多数丰富。词法并且表示分析显示在DEP2的那个变化主要影响脊柱和主要、第二等的分支的快速的延伸,但是不损害圆锥花序primordia的开始或形成。进一步的分析建议在dep2的圆锥花序长度的减少被一个缺点在圆锥花序的指数的延伸期间在房间增长引起。尽管有在dep2异种的一种更紧缩的植物类型,在谷物生产的重要改变都没在野类型和dep2异种之间被发现。因此,DEP2的学习不仅加强我们圆锥花序建筑学的分子的基因基础的理解而且为米饭繁殖有重要含意。
简介:One-cellmouseembryosfromKMstrainandB6C3F1strainwereculturedinM16medium,inwhich2-cellblockgenerallyoccurs.EmbryosofKMstrainexhibited2-cellblock,whereasB6C3F1embryos,whichareregardedasanonblockingstrain,proceededtothe4-cellstageinourculturecondition.Itisoftenassumedthattheblockofearlydevelopmentisduetothefailureofzygoticgeneactivation(ZGA)inculturedembryos.Inthisstudyweexaminedproteinsynthesispatternsbytwo-dimensionalgelelectrophoresisof[35S]methionineradiolabeled2-cellembryos.Embryosfromtheblockingstrainandthenonblockingstrainwerecomparedintheirdevelopmentbothinvitroandinvivo.ThedetectionofTRCexpression,amarkerofZGA,at42hposthCGinKMembryosdevelopedinvitrosuggestedthatZGAwasalsoinitiatedeveninthe2-cellarrestedembryos.Nevertheless,asignificantdelayofZGAwasobservedinKMstrainascomparedwithnormallydevelopedB6C3F1embryos.AttheverybeginningofmajorZGAasearlyas36hposthCG,TRChasalreadybeenexpressedinB6C3F1embryosdevelopedinvitroandKMembryosdevelopedinvivo.Butfor2-cellblockedKMembryos,TRCwasstillnotdetectableevenat38hposthCG.Theseevidencessuggestthat2-cell-blockedembryosdoinitiateZGA,andthat2-cellblockphenomenonisduenottothedisabilityininitiatingZGA,buttoadelayofZGA.
简介:Humantumornecrosisfactorα(hTNFα),apleiotropiccytokinewithactivitiesrangingfromhostdefensemechanismsininfectionandinjurytoseveretoxicityinsepticshockorotherrelateddiseases,isapromisingtargetfordrugscreening.UsingtheSELEX(systematicevolutionofligandsbyexponentialenrichment)process,weisolatedoligonucleotideligands(aptamers)withhighaffinitiesforhTNFα.Aptamerswereselectedfromastartingpoolof40randomizedsequencescomposedofabout1015RNAmolecules.RepresentativeaptamersweretruncatedtotheminimallengthwithhighaffinityforhTNFαandwerefurthermodifiedbyreplacementof2'-OHwith2'-Fand2'-NH2atallribopurinepositions.ThesemodifiedRNAaptamerswereresistanttonuclease.ThespecificityoftheseaptamersforhTNFαwasconfirmed,andtheiractivitytoinhibitthecytotoxicityofhTNFαonmouseL929cellswasdetermined.Resultsdemonstratedthatfour2'-NH2-modifiedaptamersboundtohTNFαwithhighaffinityandblockedthebindingofhTNFαtoitsreceptor,thusprotectingtheL929cellsfromthecytotoxicityofhTNFα.OligonucleotideaptamersdescribedherearepotentialtherapeuticsanddiagnosticsforhTNFc-relateddiseases.
简介:Domaindatabaseisessentialfordomainpropertyresearch.Eliminatingredundantinformationindatabasequeryisveryimportantfordatabasequality.Herewereportthemanualconstructionofanon-redundanthumanSH2domaindatabase.Thereare119humanSH2domainsin110SH2-containingproteins.HumanSH2swerealignedwithClustalX,andahomologoustreewasgenerated.Inthistree,proteinswithsimilarknownfunctionwereclassifiedintothesamegroup.Someproteinsinthesamegrouphavebeenreportedtohavesimilarbindingmotifsexperimentally.Thetreemightprovidecluesaboutpossiblefunctionsofhypotheticalproteinsforfurtherexperimentalverification.
简介:Inordertostudythemechanismoftheeffectofheparinonapoptosisincarcinomacells,thenasopharyngealcarcinomacelllineCNE2wasusedtoidentifytheeffectofheparinonapoptosisassociatedwiththeexpressionofc-myc,bax,bcl-2proteinsbyuseofHoechst33258staining,terminaldeoxynucleotidyltransferase-mediateddUTPnick-endlabeling(TUNEL),agarosegelelectrophoresis,andflowcytometry,aswellasWesternblotanalysis.TheresultsshowedthatheparininducedapoptosisofCNE2cellsincludingthemorphologicchangessuchasreductioninthevolume,andthenuclearchromatincondensation,aswellasthe“ladderpattern”revealedbyagarosegelelectrophoresisofDNAinaconcentration-dependentmanner.ThenumberofTUNEL-positivecellswasdramaticallyincreasedto33.6±1.2%from2.8±0.3%bytreatmentwithheparinindifferentconcentrations(10~40kU/L).Theapoptoticindexwasincreasedto32.5%from3.5%bydetectingSubG1peaksonflowcytometry.Westernblotanalysisshowedthatlevelsofbcl-2,baxandc-mycweresignificantlyoverexpressedbytreatmentwiththeincreaseofheparinconcentrations.TheseresultssuggestthatheparininducesapoptosisofCNE2cells,whichmayberegulatedbydifferentialexpressionofapoptosis-relatedgenes.
简介:导致死亡的肿瘤坏死的能力因素相关的导致apoptosisligand(小道)大部分被描述了有选择地杀死许多癌症房间,但是有治疗的主要担心之一是药抵抗和可能的有毒的副作用的出现。这里,我们报导那条小道在Jurkat和SUPT1T房间线并且在人的高强风然而并非在健康的导出题目的外部血mononuclear房间导致apoptosis。在平行,有小道和Tyrphostin(AG-490)的治疗,选择Januskinase2禁止者,生产cytotoxicity的明显的改进,与控制相比或到由Stat3phosphorylation的重要抑制描绘了落后于独自一个对待的样品,并且与cIAP-1和cIAP-2mRNA层次的戏剧的减少联系了。由特定的小干扰RNA的cIAP-1和cIAP-2的Downregulation显著地放大减少小道的cytotoxicity。所有一起,这些调查结果强烈显示cIAP-1和cIAP-2downregulation是在调停的发信号的小径的基本的步T上的小道和AG-490的组合效果房间白血病。这些调查结果可以帮助在小道敏感的白血病影响的病人的治疗为不太有毒的药理学策略的发展打开新线路。
简介:Apoptosismanifestsintwomajorexecutionprogramsdownstreamofthedeathsignal:thecaspasepathwayandorganelledysfunction.Animportantantiapoptosisfactor,Bcl-2protein,contributesincaspasepathwayofapoptosis.Calcium,animportantintracellularsignalelementincells,isalsoobservedtohavechangesduringapoptosis,whichmaybeaffectedbyBcl-2protein.WehavepreviouslyreportedthatinHarringtonine(HT)inducedapoptosisofHL-60cells,there'schangeofintracellularcalciumdistribution,ovingfromcytoplastespeciallyGolgi'sapparatustonucleusandaccumulatingtherewiththehighestconcentration.Wereportherethatcaspase-3becomesactivatedinHT-inducedapoptosisofHL-60cells,whichcanbeinhibitedbyoverexpressionofBcl-2protein.NosignofapoptosisorintracellularcalciummovementfromGolgi'sapparatustonucleusinHL-60cellsoverexpressingBcl-2ortreatedwithAc-DEVD-CHO,aspecificinhibitorofcaspase-3.Theresultsindicatethatactivatedcaspase-2canpromotethemovementofintracellularcalciumfromGolgi'sapparatustonucleus,andtheprocessisinhibitedbyAc-DEVD-CHO(inhibitorofcaspase-3),andthatBcl-2caninhibitthemovementandaccumulationofintracellularcalciuminnucleusthroughitsinhibitiononcaspase-3.Calciumrelocalizationinapoptosisseemstobeirreversible,whichisdifferentfromtheintracellularcalciumchangescausedbygrowthfactor.
简介:1病历摘要患者男性,25岁。因发热、咳嗽、咳痰、乏力消瘦2个月余人院。患者2006年4月1日因劳累受凉后出现发热,体温最高达39.5℃,无明显时间规律性,伴咽痛、咳嗽、咳痰,无胸痛,痰为白色黏痰,量20~30ml/d,在当地诊所静滴抗感染药(药物不详)治疗3d后仍不规律发热,咽痛好转,咳嗽、咳痰无减轻,于2006年4月17日就诊广州市某三甲医院,胸部x线片示双肺间质性病变,左上肺有肺大泡形成及纵隔数个淋巴结肿大,未行特殊治疗。为进一步诊治,患者于2006年5月11日至我院就诊,呼吸科以肺部感染收住院。查血常规白细胞6.47×10^9/L,其中淋巴细胞百分比30.8%、中性粒细胞百分比69.2%,血红蛋白117g/L,血小板331×10^9/L,纯结核蛋白衍生物PPD(-),结核抗体弱阳性,血沉82mm/h;痰未找见抗酸杆菌、真菌,普通培养无致病菌生长;
简介:【背景】沙巴拟刀角瓢虫和越南斧瓢虫是自东南亚地区引进的粉虱类害虫的优势捕食性天敌,然而其控制烟粉虱的有效性尚未明确。【方法】在室内研究了2种瓢虫幼虫捕食烟粉虱若虫时取食、爬行与休息等行为的时间分配,以及2种瓢虫幼虫和成虫在既定时间内对烟粉虱的取食时间和取食量。【结果】随着瓢虫幼虫龄期的增大,其取食烟粉虱低龄和高龄若虫的时间和休息时间均逐渐减少,而爬行寻找猎物的时间逐渐增多。沙巴拟刀角瓢虫幼虫取食烟粉虱各虫态的时间显著长于越南斧瓢虫幼虫,取食量也显著大于后者;而沙巴拟刀角瓢虫成虫取食烟粉虱的时间长于越南斧瓢虫成虫,但取食量明显低于后者。【结论与意义】2种瓢虫对烟粉虱都有较好的捕食作用,但其时间分配策略有所不同。因此,应根据田间烟粉虱的发生时期,选择合适的瓢虫进行释放。
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简介:目的:对目前最常用的检测微小RNA(miRNA)的茎环实时定量PCR法和PAP实时定量PCR法进行比较。方法:分别用茎环实时定量PCR法和PAP实时定量PCR法检测人乳腺癌细胞MCF-7中U6和23种miRNA的表达,利用定量PCR分析软件和琼脂糖凝胶电泳方法,将2种方法在引物设计难度、特异性与灵敏度,以及检测通量方面进行比较。结果:茎环法的特异性和灵敏度比PAP法高,但引物设计难度大,检测通量低;PAP法引物设计难度较低,检测通量较高,但特异性和灵敏度较差。结论:茎环法实时定量PCR适于有针对性地检测小规模miRNA,而PAP法则适于大规模miRNA筛选实验。