简介:Withtheexpansionofthegeneexpressionprofiledatabase,inthecaseofaslittleaspossibletoloseinformationortoretainthemostcriticalinformation,geneextractionhasbecomeamaindirectionforthescholars.Thispaperexcludes1561irrelevantgenesthroughthedefinitionofweighteddistancefirstly,andthenremoves252redundantgenesbyPearson'scorrelationcoefficient.Finallybycomparingthetwomethods,stepwiseregressionafterclusteringandonlystepwiseanalysis,weobtainthebestcombinationof8genes.
简介:AIM:Totransfectthecatcornealendothelialcells(CECs)withrecombinanthumanβ-nervegrowthfactorgeneadeno-associatedvirus(AAV-β-NGF)andtoobservetheeffectoftheexpressedβ-NGFproteinontheproliferationactivityofcatCECs.METHODS:TheendotheliumofcatcorneawastornunderthemicroscopeandrapidlycultivatedinDulbecco’smodifiedEagle’smedium(DMEM)toformsinglelayerCECsandthepassage2endothelialcellswereusedinthisexperiment.TherecombinanthumanAAV-β-NGFwasconstructed.TherecombinanthumanAAV-β-NGFwastransferredintocatCECsdirectly.Threegroupswereasfollowing:normalCECcontrolgroup,CEC-AAVcontrolgroupandrecombinantCECAAV-β-NGFgroup.Forty-eighthoursaftertransfection,thetotalRNAwasextractedfromtheCECbyTrizol.Theexpressionoftheβ-NGFtargetgenedetectedbyfluorescencequantitativepolymerasechainreaction;proliferationactivityofthetransfectedCECdetectedat48hbyMTTassay;thepercentageofG1cellsamongCECsaftertransfectwasdetectedbyflowcytometrymethod(FCM);cellmorphologywasobservedunderinvertedphasecontrastmicroscope.RESULTS:Thetornendotheliumculturetechniquerapidlycultivatedsinglelayercatcornealendothelialcells.Theself-designedprimersforthetargetgeneandreferencegenewereefficientandspecialconfirmedthroughelectrophoresisanalysisandDNAsequencing.Forty-eighthoursaftertransfect,thehumanβ-NGFgenemRNAdetectedbyfluorescencequantitativepolymerasechainreactionshowedthattherewasnosignificantdifferencebetweennormalCECcontrolgroupandCECAAVcontrolgroup(P>0.05);therewassignificantdifferencebetweentwocontrolgroupsandrecombinantCEC-AAV-β-NGFgroup(P<0.05).MTTassayshowedthattransfectofrecombinantAAV-β-NGFpromotedtheproliferationactivityofcatCEC,whiletherewasnosignificantdifferencebetweennormalCECcontrolgroupandCEC-AAVcontrolgroup(P>0.05).FCMresultshowedthatthepercentageofG1cellsinCECAAV-NGFgroupwas76.8%whi
简介:为增加的目的在里面polycation基因搬运人的vivo稳定性,我们准备了一种pH敏感poly(乙烯乙二醇)-poly(-benzyl-L-glutamate-co-glutamic酸)(木钉针网阵列(65),65表示glutamic酸的臼齿的比率在poly(-benzyl-L-glutamate-co-glutamic酸))。木钉针网阵列(65)显示出低cytotoxicity并且能高效地防护DNA/PEI(1:1)polyplexes的正电荷。transfection被提高由于部分收费在7.4的pH在HeLa房间线防护。有几乎没有transfection效率当第三的粒子的表面费用转到时在7.4的pH否定。然而,transfection效率在transfection的开始在6.0的pH由culturing恢复了很多。共焦的显微镜的观察和流动cytometry结果证明DNA/PEIpolyplexes应该高效地被释放并且在pH的endocytosized6.0,因为pH木钉针网阵列(65)的触发的deshielding行动。由于好biocompatibility和合适的pH触发了shielding/deshielding性质,木钉针网阵列(65)能是为在vivo使用的polycationic基因搬运人的一个潜在的防护系统。
简介:目的分析一个大前庭水管综合征家系的临床特征和SLC26A4基因检测特点。方法对一个大前庭水管综合征家系进行病史采集和听力学检测,绘制耳聋家系系谱图,提取受检者的基因组DNA,对所有家系成员进行耳聋基因芯片分析和SLC26A4基因全外显子及外显子侧翼序列的检测。结果该家系共5代合计30人(男17人,女13人),现存26人,其中第四代7人为耳聋患者,6人均为语前感音神经性聋。1人为语后感音神经性聋,颞骨CT显示均为前庭水管扩大,第五代1人为耳聋患者,表现为前庭水管扩大合并语前感音神经性聋。在家系中共发现SLC26A4基因c.754C〉T、c.919-2A〉G、c.1264-12T〉A和c.1548_1549insC四种已知致病突变类型。耳聋患者均为复合杂合突变。10人为SLC26A4基因携带者。结论该家系的8例耳聋患者由SLC26A4基因复合杂合突变导致前庭水管扩大,病因学分析可为患者提供预见性的临床措施,并为该家系的后代遗传咨询与婚育指导提供理论依据及科学手段,有效地防止聋儿后代出生。
简介:设计习俗的核酸酶技术例如定期聚类短palindromic重复(CRISPR)联系了的interspaced(Cas)系统提供为昆虫编辑工具的吸引人的染色体功能的遗传。指向的基因mutagenesis由CRISPR/Cas9系统调停了在包括的几份昆虫订单被完成了双翅目,鳞翅目和翘目。然而,小成功都没在这些由于genomic信息和胚胎的microinjection技术的缺乏在农业害虫被报导昆虫种类。这里,我们报导CRISPR/Cas9系统在重要鳞翅类的害虫Spodopteralitura导致了有效基因mutagenesis。我们指向了S。litura腹A(Slabd--一)是重要胚胎的发展基因并且在决定昆虫的腹的片断的身份起一个重要作用的基因。指导Cas9送信人RNA和Slabd-A-specific单人赛领队RNA(sgRNA)的注射进S。成功地导致的litura胚胎典型abd--缺乏的显型,它在幼虫的阶段期间显示出异常分割和宫外的色素沉着。聚合酶链基于反应的分析表明Cas9/sgRNA建筑群有效地在S导致了指向的mutagenesis。litura。这些结果证明CRISPR/Cas9系统是为在象S那样的鳞翅类的害虫的染色体操作的一个强大的工具。litura。
简介:丝胶是在Bombyxmori的中间的丝绸腺明确地表示的一种水溶性的蛋白质。当sericin-1基因倡导者被克隆,转基因的向量被构造表示外国蛋白质时,特定的电子螺旋管,Bmhel-8,在Bombyxmori和Bombyxmandarina的一些遗传型在sericin-1基因倡导者顺序被识别。给Bmhel-8电子螺旋管transposon在一些遗传型仅仅是在场的,它能是在sericin-1倡导者的突变而产生之遗传的变化的来源。sericin-1倡导者顺序的长度是约1063或643bp。顺序或等位基因的更大的尺寸被归功于到Bmhel-8的存在。蚕遗传型能为也是同型结合的更矮或更大的倡导者顺序或异质接合,包含两等位基因。在sericin-1倡导者的Bmhel-8展出enhancer活动,在BmE房间线由一个双酶的记者系统示威了。而且,Bmhel-8在sericin-1显示enhancer活动倡导者驱动的基因表达式系统但是不调整sericin-1的织物特定的表达式。
简介:Background:Thoughthemechanismsofskeletalmuscleregenerationaredeeplyunderstood,thoseinvolvedinmusclecontusion,oneofthemostcommonmuscleinjuriesinsportsmedicineclinics,arenot.Theobjectiveofthisstudyistoexplorethemechanismsinvolvedinmuscleregenerationaftercontusioninjury.Methods:Inthisstudy,atotalof72micewereused.Eightofthemwererandomlychosenforthecontrolgroup,whiletherestweresubjectedtomusclecontusion.Subsequently,theirgastrocnemiusmuscleswereharvestedatdifferenttimepoints.Thechangesinmusclemorphologywereassessedbyhematoxylinandeosin(HE)stain.Inaddition,thegeneexpressionwasanalyzedbyreal-timepolymerasechainreaction.Results:Thedatashowedthattheexpressionofmanygenes,i.e.,specificmarkersofimmunecellsandsatellitecells,regulatoryfactorsformuscleregeneration,cytokines,andchemokines,increasedintheearlystagesofrecovery,especiallyinthefirst3days.Furthermore,therewerestrictrulesintheexpressionofthesegenes.However,almostallthegenesreturnedtonormalat14dayspost-injury.Conclusion:Thesequenceofimmunecellsinvadedaftermusclecontusionwasneutrophils,M1macrophagesandM2macrophages.SomeCC(CCL2,CCL3,andCCL4)andCXC(CXCL10)chemokinesmaybeinvolvedinthechemotaxisoftheseimmunecells.HGFmaybetheprimaryfactortoactivatethesatellitecellsaftermusclecontusion.Moreover,2weeksareneededtorecoverwhenacutecontusionhappensasusedinthisstudy.
简介:摘要目的探讨儿童肾癌的特点、诊断、治疗和预后。方法回顾性分析了自2005~2014年我院收治7例肾癌患儿,并进行相关文献复习。结果7例中男性患儿5例,女性2例,平均年龄10.1岁;左侧3例,右侧4例。4例因血尿就诊,2例因腹痛就诊,1例因外伤后B超发现肾脏占位。7例Robson分期均在I期~II期内,其中4例为Xp11.2易位/TFE3基因融合相关性肾癌。随访3月~8年,未见肿瘤复发或转移,生存率100%。结论肾癌在儿童较少见。诊断上以B超结合CT或MRI检查明确诊断,需与肾母细胞瘤鉴别。治疗上采用开放或腹腔镜下瘤肾切除术,术中行淋巴结活检。术后患儿的预后与肿瘤分期有一定关系。
简介:重点词汇1.papern.纸;纸张pape袁示'纸;纸张'时,是不可数名词。apieceofpaper一张纸拓展(1)paper可作定语,如:papermoney纸币。(2)paper意为'试卷;论文'时,是可数名词。Pleasehandoutthepapers.请分发试卷。2.pollutev.污染Diditpollutetheenvironment?它对环境有污染吗?拓展(1)pollution月.污染;污染物air/noisepollution空气/噪音污染(2)pollutedadj.被污染的pollutedwater被污染了的水3.buildv.建筑;建造