简介:目的:探讨1个凝血因子Ⅹ(FⅩ)缺陷症家系的分子发病机制。方法:对先证者及家系成员凝血、抗凝及纤溶功能筛查以及凝血因子活性及抗原含量检测进行表型诊断;以WesternBlotting检测血浆中FⅩ抗原含量和分子量大小;以中和试验检测FⅩ的抑制物。以PCR方法对F10基因所有外显子及侧翼序列和5’端非翻译区进行扩增,产物纯化后直接测序进行基因诊断;构建F10基因突变表达质粒,瞬时转染HEK293T细胞,测定表达产物的FⅩ促凝活性(FⅩ:C)和FⅩ抗原含量(FⅩ:Ag)。结果:先证者FⅩ:C和FⅩ:Ag分别为〈1%和53.36%,中和试验结果阴性,诊断为交叉反应物质阳性(CRM+)的FⅩ缺陷症。F10基因分析发现2个杂合突变:IVS5+1G〉A和Asp368del。Asp368del体外表达显示FⅩ:C和FⅩ:Ag分别为(0.52±0.04)%和(85.9±5.0)%,为CRM+突变。结论:F10基因双重杂合突变IVS5+1G〉A和Asp368del导致该家系遗传性FⅩ缺陷症。剪接位点突变IVS5+1G〉A导致内含子无法正常剪接,影响FⅩ正常表达。Asp368del突变蛋白能够正常表达,但功能降低。
简介:Objective:Fusogenicendogenousretroviralsyncytinplaysanimportantroleintheformationofsyncytiotrophoblastsinhumanplacenta.Apartfromitsexpressioninplacenta,brainandtestis,syncytinhasalsobeenfoundinmanycancers.Althoughsyncytinhasbeenproposedtoserveasapositiveprognosticmarkerinsomecancers,theunderlyingmechanismisunclear.Theaimofthisstudyistoevaluatetheeffectsofsyncytinexpressionontheinvasivephenotypeofmelanomacells.Methods:Theeukaryoticexpressionplasmidforsyncytin-EGFPwasconstructedandtransfectedintoB16F10melanomacells.TheeffectofsyncytinontheinvasionpotentialoftumorcellswasevaluatedinB16F10sublinecellsthatstablyexpressedsyncytin-EGFPfusionproteinorEGFPalone.Results:TheB16F10sublinesthatstablyexpressedsyncytin-EGFPorEGFPalonewereestablishedrespectivelyandconfirmedbyimmunofluorescentandimmunoblottingassay.SyncytinexpressioninB16F10cellswasassociatedwithdecreasedcellproliferation,migrationandinvasion.Multinucleatedgiantcellsthatcontainedasmanyasfivenucleiwereinducedinsyncytin-expressingcells.Inaddition,syncytinexpressiondidnotalterthesensitivityofB16F10cellstotrichosanthin,atoxinthatdamagessyncytiotrophoblastsmoreefficientlythanothertissues.Conclusions:Theseresultssuggestthatsyncytinexpressioninsomecancersmayconfinetheirinvasionpotentialandthusserveasapositiveprognosticfactor.更多还原
简介:介绍了一种适用于微细金属线恒张力放线的主动放线机的结构原理,详细论述了以单片机PIC18F66J10为主控芯片,以SLA7026为步进电机驱动芯片的硬件电路设计和以PI控制算法为主的系统软件设计方法。