简介：Thec-erbB-2proto-oncogeneencodesa185kDaproteinp185,whichbelongstoepidermalgrowthfactorreceptorfamily.Amplificationofthisgenehasbeenshowntocorrelatewithpoorclinicalprognosisforcertaincancerpatients.ThemonoclonalantibodyA21whichdirectedagainstp185specificallyinhibitsproliferationoftumorcellsoverexpressingp185,henceallowsittobeacandidatefortargetedtherapy.InordertoovercomeseveraldrawbacksofmurineMAb,wecloneditsVHandVLgenesandconstructedthesingle-chainFv(scFv)throughapeptidelinker.TherecombinantscFvA21wasexpressedinEscherichiacoliandpurifiedbytheaffinitycolumn.SubsequentlyitwascharacterizedbyELISA,Westernblot,cellimmunohistochemistryandFACS.Alltheseassaysshowedthebindingactivitytoextracellulardomain(ECD)ofp185.BasedonthosepropertiesofscFvA21,wefurtherconstructedthescFv-Fcfusionmoleculewithahomodimerformandtherecombinantproductwasexpressedinmammaliancells.Inaseriesofsubsequentanalysisthisfusionproteinshowedidenticalantigenbindingsiteandactivitywiththeparentantibody.Theseanti-p185engineeredantibodieshavepromisedtobefurthermodifiedasatumortargetingdrugs,withaviewofapplicationinthediagnosisandtreatmentofhumanbreastcancer.

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简介：Amurinemacrophage-likecelllineJ774,acquired,inresponsetoLPS,anabilitytokilltumornecrosisfactor(TNF)-insensitivetargetP815mastocytomacellswhereasanothercellline,P388D1didnot,LPStriggeredsignalingmechanismsbetweenthetwocelllineswerecomparedwithanaimtoinquireaboutthepossiblenatureoftheabove-mentioneddifference,TheresultswhowedthattwocelllinesrespondtoLPS-treatmentbyparallelactivationofbothphospholipasesCandA2(PLCandPLA2)toapproximatelythesameextent.ThemaximumresponseoftothenzymesofJ774cellswasnotedwithin10minthetreatmentwhereasthatofP388D1cellsrequiredmorethan20min,TheotherpropertiesofLPS-responsiveenzymesstudiedweresimilarbetweentwocelllines,includingActivationofPLCandPLA2andPKCinmacrophagesbyLPS.Ca2+augmentationofenzymeactivation,participationofguaninenucleotidebinding(G)proteinsintheinitialactivationpreocesses,andinhibitionofenzymeactivationbythepriortreatmentofcellswithcholeraorpertussistoxinsetc.Moreover,LPS-triggeredactivationofPLCandPLA2wasfoundtobefollowedbytheincreaseofPKCactivitiesinbothcelllines.Inspiteofthesesimilarities.J774cellspossessedbothbasicandacidicformsofPKCactivities,whileP388D1cellsownedonlyPKCofbasicform,Nevertheless,thequestionwhyJ774cellsbutnotP388D1cells,canacquirethetumoricidalactivity,aganistP815,cellsfollowingLPStreatmentrematinstobeanswered.

简介：小道，肿瘤坏死因素相关的导致apoptosisligand，是一个新奇有势力通过房间表面死亡受体Trail-R1和Trail-R2的激活的房间死亡小径的内长的使活跃之物。它的角色象在导致激活的房间死亡(AICD)的FasL一样，在免疫系统被表明了。然而，小道的机制导致了apoptosis遗体不清楚。在这份报告，重组体小道蛋白质被表示并且净化。导致apoptosis活动和JurkatT房间上的重组体小道的规定机制是探索试管内。Trypan蓝排除试金证明重组体小道蛋白质活跃地以一种剂量依赖者方式杀死了JurkatT房间。在JurkatT房间的导致小道的apoptosis被Bcl-2显著地在Bcl-2基因transfected房间在表示上减少。有PMA(phorbol12十四酸盐13醋酸盐)的处理，PKC使活跃之物，在JurkatT房间的压制的导致小道的apoptosis。由PMA的apoptosis的抑制被预告的处理与二度废除，一个PKC禁止者。总起来说，Bcl-2在表示上和PMA激活PKC，这被建议活跃地下面调整在JurkatT的调停小道的apoptosis房间。

简介：TRAF2isacriticaladaptormoleculeforTNFreceptorsininflammatoryandimmunesignaling.Uponreceptorengagement,TRAF2isrecruitedtoCD40andtranslocatestolipidraftsinaRINGfinger-dependentprocess,whichenablestheactivationofdownstreamkinases.TRAF1candisplaceTRAF2andCD40fromraftfractions,anditpromotestheabilityofTRAF2tosustainsignalactivation.ReplacementoftheRINGfingerofTRAF2witharaft-targetingsignalrestoresJNKactivationandassociationwiththecytoskeletalproteinFilamin,butnotNF-KBactivation.TRAF1-/-dendriticcellsshowattenuatedresponses

简介：<正>Usingsubtractioncloning,weidentifiedthehumanN-MycDownstream-RegulatedGene-2(hNDRG2),locatedat14q11.2,asacandidatetumorsuppressorgene.Semi-quantitativeRT-PCRshowedthattheexpressionofhNDRG2in15of27(56%)humanGBMtissuesandall6humanglioblastomacelllineswassignificantlylowerthanthatinthenormalbrain.TheexpressionofhNDRG2alsowasevaluatedin60lung-carcinomapatients.17of26casesofsquamouscarcinomaand4of11casesofsmallcelllungcancerdisplayed

简介：GelatinaseA(MMP-2)isconsideredtoplayacriticalroleincellmigrationandinvasion.Theproteinaseiscercetedfromthecellasaninactivezymogen.InvivoitispostulatedthatactivationofprogelationaseA(proMMP-2)takesplaceonthecellsurfacemediatedbymembrane-typematrixmetalloproteinases(MT-MMPs).RecentstudieshavedemonstratedthatproMMP-2isrecruitedtothecellsurfacebyinteractingwithtissueinhibitorofmetalloproteinases-2(TIMP-2)boundtoMT1-MMPbyformingaternarycomplex.FreeMT1-MMPcloselylocatedtotheternarycomplexthenactivatesproMMP-2onthecellsurface.MT1-MMPisfoundinculturedinvasivecancercellsattheinvadopodia.TheMT-MMP/TIMP-2/MMP-2systemthusprovideslocalizedexpressionofproteolysisoftheextracellularmatrixrequiredforcellmigration.

简介：在哺乳动物的胚胎，内部房间团(ICM)的分离和trophectoderm(TE)的第一种房间命运选择，，被抄写因素，Oct4和Cdx2的互相对抗的效果调整pluripotency因素，Nanog，是必要的指定epiblast。我们分析了Nanog和Cdx2的倡导者，并且发现了这二个抄写因素同样相互地被调整。用有有条件的TE区别的一根胚胎的干细胞线，我们证明Nanogoverexpression压制TE标记的upregulation，当时Nanog击倒的upregulatesTE标记的表示。我们推进Nanog和Cdx2绑在并且镇压的表演对方的倡导者。而Nanog大美人在ICM导致可检测的Cdx2表示，我们不管多么不观察胚囊开发的公开混乱，显示Nanog起到在ICM和TE的分离的Oct4的一个谄媚的作用。

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简介：Amurinemacrophage-likecellline,J774,acquried,inresponsetoLPS,anabilitytokilltumornecrosisfactor（TNF）-insensitivetargetP815mastocytomacells,whereasanothercellline,P388D1,didnot.LPS-triggeredsignalingmechanismsbetweenthetwocelllineswerecomparedwithanaimtoinquireaboutthepossiblenatureoftheabove-mentioneddifference.TheresultsshowedthattwocelllinesrespondtoLPS-treatmentbyparallelactivationofbothphospholipasesCandA2（PLCandPLA2）toapproximatelythesameextent.ThemaximumresponseofbothenzymesofJ774cellswasnotedwithin10minofthetreatment,whereasthatofP388D1cellsrequiredmorethan20min.TheotherpropertiesofLPS-responsiveenzymesstudiedweresimilarbetweentwocelllines,ineludingActivationofPLCandPLA2andPKCinmacrophagesbyLPSCa2+augmentationofenzymeactivation,participationofguaninenucleotidebinding（G）proteinsintheinitialactivationprocesses,andinhibitionofenzymeactivationbythepriortreatmentofcellswithcholeraorpartussistoxinsetc.Moreover,LPS-triggeredactivationofPLCandPLA2wasfoundtobefollowedbytheincreaseofPKCactivitiesinbothcelllines.Inspiteofthesesimilarities,J774cellspossessedbothbasicandacidicformsofPKCactivities,whileP388D1cellsownedonlyPKCofbasicform.Nevertheless,thequestionwhyJ774cells,butnotP388D1cells,canacquirethetumoricidalactiyity,aganistP815cellsfollowingLPS-treatmentremainstobeanswered.

简介：ErbB2,amemberofthereceptortyrosinekinasefamily,isfrequentlyover-expressedinbreastcancer.ProteolysisoftheextracellulardomainofErbB2resultsinconstitutiveactivationofErbB2kinase.RecentstudyreportedthatErbB2isfoundinthenucleus.Here,weshowedthatErbB2isimportedintothenucleusthroughanuclearlocalizationsignal(NLS)-mediatedmechanism.TheNLSsequenceKRRQQKIRKYTMRR(aa655-668)containsthreeclustersofbasicaminoacidsanditissufficienttotargetGFPintothenucleus.However,mutationinanybasicaminoacidclusterofthisNLSsequencesignificantlyaffectsitsnuclearlocalization.Furthermore,itwasfoundthatthisNLSisessentialforthenuclearlocalizationofErbB2sincetheintracellulardomainofErb2lackingNLScompletelyabrogatesitsnucleartranslocation.Takentogether,ourstudyidentifiedanovelnuclearlocalizationsignalandrevealsanovelmechanismunderlyingErbB2nucleartraffickingandlocalization.

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：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.

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简介：Polyamines在在植物，而是他们的准确角色调整各种各样的发展过程被含有并且他们怎么管理这些过程尚待逃犯。我们这里报导浓密的Arabidopsis和矮子异种，bud2，哪个从编码S-adenosylmethioninedecarboxylases(SAMDC)的小基因家庭的一个成员的完全的删除的结果为在polyamine简历的不可缺少的中介的形成必要合成小径。bud2植物在开花期，根，和叶柄扩大了脉管的系统，并且polyamines的改变的动态平衡。bud2和samdc1的双异种，另一个SAMDC成员的击倒的异种，是胚胎致命，证明SAMDC为植物胚胎开始是必要的。我们的结果建议polyamines为更高的植物的正常生长和发展被要求。

简介：MicroRNAs是否定地在post-transcriptional水平调制基因表示的短规章的RNA，并且深深地涉及癌症的几种类型的致病。调查特定的miRNAs和他们的目标基因是否参予喉的癌的分子的致病，oligonucleotidemicroarrays被用来与正常纸巾相比在喉的癌纸巾估计microRNAs和mRNAs的微分表示侧面。oncogenicmiRNA，microRNA-21(miR-21)，被发现是在喉的癌纸巾的upregulated。由特定的antisenseoligonucleotides的miR-21击倒而miR-21的overexpression提高了细胞的生长活动，由殖民地形成试金检测了，禁止了HEp-2细胞的增长潜力。miR-21抑制引起的房间数字减小由于G1-S阶段转变的控制的损失，而不是apoptosis的显著增加。随后，miR-21的新目标基因，BTG2，被发现是在喉的癌纸巾的downregulated。BTG2被知道充当一个平底锅房间周期管理者和肿瘤suppressor。这些调查结果显示miR-21的那异常表情可以由维持BTG2的底层贡献喉的癌的恶意的显型。oncogenicmiR-21和它的目标基因的鉴定，BTG2，为癌症诊断和治疗在喉的癌是潜在地珍贵的。