简介:Objective:AllelicpolymorphismsofCCR5△32、CCR2b-64I,CX3CR1-249I280MandSDF1-3’AassociatedwithHIV-1infectionanddiseaseprogressionwereinvestigatedinindigenousUygurpopulationsfromtheXinjiangUygurAutonomousRegionofChina.Mithods:Thestudypopulationcomprised316healthyUygursubjectswithanagerangeof1-80yearsold,fromwhomwholeperipheralbloodsampleswerecollectedandnonewereHIV-1seropositive.GenomicDNAsampleswerepurifiedusingaQiagenBloodKit.GenotypingoftheaforementionedfouralleleswasperformedusingPCRorPCR/RFLPassay,andfurtherconfirmedbydirectDNAsequencing.Results:TheallelicfrequenciesinChineseUygurpopulationwereasfollows:3.48%forCCR5△32;19.45%forCCR2b-64I;13.8%forCX3CRI-249I280Mhaplotype,and20.41%forSDFI-3’A.MutantalleledistributionsamongUygurpopulationswereinaccordancewiththeHardy-Weinbergequilibrium.NostatisticaldifferencewasfoundbetweenthefrequencyofthethreeHIVcoreceptorsandtheirrespectiveligandgenes.Conclusion:ThefrequencyofSDF1-3’AandCX3CR1-249I280MhaplotypemorecloselymatchedthehanChinese.ThefequencyofCCR5△32inUygurpopulationswasbetweenCaucasianandHanfrequencies,themorecloselymatchingthefrequencyinMedi-Asiapeople.NogeneticlinkagebetweenanytwoofthethreeHIVcoreceptorgeneswasfound,butobviousgeneticlinkagesexistedbetweenCX3CR1-249IandCX3CR1-280M,withevenhigherlinkagedegreesthanCaucasianpeople.
简介:内容摘要 :中国标准动车组项目开关柜项目开关柜线束数量大、元器件多,提升开关柜内线束施工标准是提高其标准化、规范化及美观程度的重要手段。根据车辆各接线点实际位置,通过反复改良分线码线路径及施工细节,制定出开关柜线束分线、码线及整体恢复的施工标准。该标准在原有的施工要求基础上对每一施工步骤做出具体数据明确,实现了开关柜电气线束的施工标准的统一。
简介:目的:构建带Myc标签的LC3B—PLA2基因的真核表达质粒,获得LC3B—PLA2融合蛋白,并应用LC3B—PLA2研究Atg4B对LC3B的切割作用。方法:以本实验室保存的乳腺文库为模板,PCR扩增获得LCgB序列,与PLA2G]O拼接后插入pCMV—Myc载体,用构建的重组质粒转染HEK293T细胞,Western印迹检测融合蛋白的表达,用LC3B—PLA2与Atg4B共转染的方法检测Atg4B对LC3B的切割作用。结果:菌液PCR、重组质粒双酶切及测序均表明重组质粒构建成功,Western印迹结果表明融合蛋白在HEK293T细胞中获得表达,LC3B—PLA2真核表达蛋白能够应用于Atg4B对LC3B切割作用的研究。结论:构建了pCMV—Myc—LC3B—PLA2真核表达质粒,LC3B—PLA2融合蛋白在Atg4B对LC3B切割作用的研究中至关重要,为进一步研究Atg4B在自噬过程中的作用奠定了基础。
简介:MicroRNAs(miRNAs)aresmall,non-codingRNAsthatnegativelyadjustgeneexpressioninmultifariousbiologicalprocesses.However,theregulatoryeffectsofmiRNAsonSchwanncellsremainpoorlyunderstood.PreviousmicroarrayanalysisresultshaveshownthatmiRNAexpressionisalteredfollowingsciaticnervetransaction,therebyaffectingproliferationandmigrationofSchwanncells.ThisstudyinvestigatedwhethermiR-148b-3pcouldregulatemigrationofSchwanncellsbydirectlytargetingcullin-associatedandneddylation-dissociated1(Cand1).Up-regulatedexpressionofmiR-148b-3ppromotedSchwanncellmigration,whereassilencingofmiR-148b-3pinhibitedSchwanncellmigrationinvitro.FurtherexperimentsconfirmedthatCandlwasadirecttargetofmiR-148b-3p,andCandlknockdownreversedsuppressionofthemiR-148b-3pinhibitoronSchwanncellmigration.TheseresultssuggestedthatmiR-148b-3ppromotedmigrationofSchwanncellsbydirectlytargetingCandlinvitro.
简介:摘要目的研究miR-125b-1-3p在轮状病毒(rotavirus)复制过程中的作用和调控机制。方法将MA104细胞中的miR-125b-1-3p分别上调和下调后感染轮状病毒,通过RT-PCR分析miR-125b-1-3p的表达情况和轮状病毒拷贝数;免疫荧光分析miR-125b-1-3p对轮状病毒蛋白表达情况的影响;Western blot分析miR-125b-1-3p参与调控的相关蛋白表达情况。结果轮状病毒感染细胞后,miR-125b-1-3p的表达水平明显上调,上调miR-125b-1-3p后,轮状病毒的VP7、NSP3基因拷贝数下降,下调miR-125b-1-3p后,轮状病毒的VP7、NSP3基因拷贝数明显升高;上调miR-125b-1-3p的表达水平后轮状病毒蛋白荧光数下降,下调miR-125b-1-3p后,病毒蛋白荧光数增加;轮状病毒感染细胞16 h后PI3K/Akt通路活性受到抑制,上调miR-125b-1-3p能够抑制PI3K/Akt通路的活化。结论miR-125b-1-3p通过调控PI3K/Akt通路抑制轮状病毒的复制。研究结果为探讨miR-125b-1-3p和PI3K/Akt通路之间的具体调控机制研究提供了实验基础,为轮状病毒抗感染治疗提供了靶点。
简介:AbstractStroke is a devastating disease that occurs when a blood vessel in the brain is either blocked or ruptured, consequently leading to deficits in neurological function. Stroke consistently ranked as one of the top causes of mortality, and with the mean age of incidence decreasing, there is renewed interest to seek novel therapeutic treatments. The Scavenger Receptor Class B type 1 (SR-B1) is a multifunctional protein found on the surface of a variety of cells. Research has found that that SR-B1 primarily functions in an anti-inflammatory and antiatherosclerotic capacity. In this review, we discuss the characteristics of SR-B1 and focus on its potential correlation with the modifiable risk factors of stroke. SR-B1 likely has an impact on stroke through its interaction with smoking, diabetes mellitus, diet, physical inactivity, obesity, hypercholesterolemia, atherosclerosis, coronary heart disease, hypertension, and sickle cell disease, all of which are critical risk factors in the pathogenesis of stroke.
简介:摘要:目的 探讨在膀胱癌中MiR-29b-3p靶向调节COL1A1的表达及临床意义。方法 利用Oncomine数据库对COL1A1在膀胱癌组织与正常组织、表浅性膀胱癌组织与浸润性膀胱癌组织中的表达进行差异分析;并使用cBioPortal数据库分析COL1A1与患者总体生存期的关系;TCGAportal数据库分析高表达与低表达COL1A1患者之间的生存曲线,了解预后情况,同时分析COL1A1基因表达水平与膀胱癌分期、膀胱癌分级的相关性。数据库预测microRNA并通过OncomiR数据库分析microRNA表达水平及与膀胱癌患者临床数据相关性。结果 COL1A1在膀胱癌中高表达,随着分级及分期的增加,COL1A1的表达量递增;生存分析显示,COL1A1高表达的膀胱癌患者预后不佳,生存时间缩短;通过数据库预测COL1A1上游miRNA MiR-29b-3P的证据最强;生存分析显示,MiR-29b-3P高表达的膀胱癌患者预后更佳,生存时间延长。结论 COL1A1在膀胱癌组织中呈高表达,其高表达患者预后较差,MiR-29b-3P高表达患者预后更佳,生信分析表明膀胱癌中MiR-29b-3p靶向调节COL1A1表达抑制膀胱癌进展,为进一步实验验证奠定研究基础,可能作为膀胱癌治疗靶点的潜力。
简介:AbstractBackground:Long non-coding RNA (lncRNA) actin filament-associated protein 1 antisense RNA 1 (AFAP1-AS1) functions as a competing endogenous RNA to regulate target genes expression by sponging microRNAs (miRs) to play cancer-promoting roles in cancer stem cells. However, the regulatory mechanism of AFAP1-AS1 in cervical cancer (CC) stem cells is unknown. The present study aimed to provide a new therapeutic target for the clinical treatment of CC.Methods:Hyaluronic acid receptor cluster of differentiation 44 variant exon 6 (CD44v6)(+) CC cells were isolated by flow cytometry (FCM). Small interfering RNAs of AFAP1-AS1 (siAFAP1-AS1) were transfected into the (CD44v6)(+) cells. The levels of AFAP1-AS1 were measured by quantitative real-time PCR (qRT-PCR). Sphere formation assay, cell cycle analysis, and Western blotting were used to detect the effect of siAFAP1-AS1. RNA pull-down and luciferase reporter assay were used to verify the relationship between miR-27b-3p and AFAP1-AS1 or vascular endothelial growth factor (VEGF)-C.Results:CD44v6(+) CC cells had remarkable stemness and a high level of AFAP1-AS1. However, AFAP1-AS1 knockdown with siAFAP1-AS1 suppressed the cell cycle transition of G(1)/S phase and inhibited self-renewal of CD44v6(+) CC cells, the levels of the stemness markers octamer-binding transcription factor 4 (OCT4), osteopontin (OPN), and cluster of differentiation 133 (CD133), and the epithelial-mesenchymal transition (EMT)-related proteins Twist1, matrix metalloprotease (MMP)-9, and VEGF-C. In the mechanism study, miR-27b-3p/VEGF-C signaling was demonstrated to be a key downstream of AFAP1-AS1 in the CD44v6(+) CC cells.Conclusions:LncRNA AFAP1-AS1 knockdown inhibits the CC cell stemness by upregulating miR-27b-3p to suppress VEGF-C.
简介:摘要:目的:分析COL1A1在膀胱癌及癌旁组织中表达差异,研究COL1A1下调抑制膀胱癌细胞增殖和迁移的机制。方法:采用实时荧光定量 PCR检测 8例膀胱癌组织、癌旁组织和膀胱癌细胞系T24、UM-UC-3,以及人正常膀胱上皮细胞系SV-HUC-1中COL1A1 mRNA及预测miRNA表达水平;敲减COL1A1和阴性对照转染T24,通过 CCK-8、Transwell实验检测细胞的增殖、迁移水平;生物信息学预测COL1A1上游miRNA, miRNA-29b-3p mimic转染 T24细胞后实时荧光定量 PCR检测检COL1A1 mRNA表达水平;采用双荧光素酶实验验证miRNA-29b-3p和 COL1A1之间的关系。结果:与癌旁组织相比,COL1A1在膀胱癌组织中表达明显升高。COL1A1敲减后,明显抑制 T24增殖、迁移;生信预测miRNA-29b-3p与COL1A1靶向结合证据最强;与癌旁组织相比,miRNA-29b-3p在膀胱癌组织中表达明显降低,miRNA-29b-3p过表达之后,COL1A1 mRNA表达水平明显降低。双荧光素酶报告实验显示,miRNA-29b-3p靶向 COL1A1 mRNA的 3'非编码区。结论:相较于癌旁组织,COL1A1在膀胱癌组织中高表达,miRNA-29b-3p在膀胱癌组织中低表达;miRNA-29b-3p通过靶向下调COL1A1实现抑制膀胱癌进展,从而降低膀胱癌细胞的增殖、迁移。