简介:目的构建并筛选出最有效的HPV16E6基因特异的小干扰RNA(smallinterferingRNA,siRNA)表达载体,观察其对宫颈癌细胞中HPV16E6基因表达的长期影响,探讨E6基因在宫颈癌发生过程中的分子作用机制,为临床HPV感染及宫颈癌治疗探索新方法.方法构建HairpinsiRNA质粒,稳定转染宫颈癌SiHa细胞,鉴定转染细胞中的质粒DNA,通过Real-TimeRT-PCR检测细胞中HPV16E6mRNA表达,采用Western-blot检测p53、p21等蛋白的变化.MTT法(四甲基偶氮唑盐微量酶反应比色法)检测SiHa细胞转染siRNA后细胞增殖曲线.结果HPV16E6AhairpinsiRNA表达载体转染人宫颈癌SiHa细胞,可以在细胞内长期表达siRNA,有效抑制细胞内HPV16E6基因的表达.E6AsiRNA能抑制细胞生长,作用持续达4个月以上.结论利用siRNA表达载体抑制整合在细胞中的外源HPVE6病毒癌基因可能是治疗HPV感染和宫颈癌的一种新的理想方法.
简介:ToleamwhetherHPV16E6/E7genescorrelatewithcervicalcarcinomaandprecancerouslesions.MethodsAmplificationandcloningofHPV16E6/E7geneswereperformedbyPCRandmolecularbiologicaltechniquesfortheirsequences.Cervicalcancer,CINⅡ-Ⅲandcervicitiswereconfirmedbypathologicdetections.ResultsDetectionratesofHPV16E6/E7inbiopsiesofcervicalcarcinoma,CINⅡ-Ⅲandcervicitiswereindividually70%,75%and65%.StatisticresultsshowthatthereisnodifferenceamongtheminHPV16E7detection.AlsotherisnodifferencebetweencervicalcancerandCINⅡ-ⅢinHPV16E6detection,however,bothwerehigherdetectionsthanincervicitisstatistically.ConclusionHPV16E6/E7genescorrelatewithcervicalcarcinomaandprecancerouslesions.E6genemightespeciallyfunctiononoccurrenceofcervicalcarcinomafromprecancerouslesions.
简介:牙科手机在操作时需要伸到患者口腔内,接触患者的唾液、血液等体液,成为细菌、病毒传播的重要媒介。由于牙科手机是精密仪器,结构复杂,任何表面消毒方法均不能起到有效的灭菌作用,为了确保无菌,防止交叉感染,2001年5月我科开始选择了高温高压蒸气灭菌法,使用EURONDA(E6-18)全自动消毒炉对手机进行彻底灭菌,施行“一人一机一高压灭菌”,取得了满意效果,现介绍如下。
简介:目的:观察抗氧化剂维生素E对慢性肾衰这残存肾组织和残肾功能的保护作用。方法:在5/6肾切除大鼠慢性肾衰模型上,以假手术大鼠为对照,采用形态计量和生化测定方法,对比分析术后30,60,90,120d给予或不给予维生素E治疗的慢性肾衰大鼠残肾纤维化程度,残肾功能及其活性氧化代谢状况等指标。结果:大鼠术后残存肾小球呈代偿性肥大,肾小球毛细血管代谢性增生的同时,有显著的氧自由基反应亢进,抗氧化能力减弱,肾小球硬化,滤过膜增厚,肾间质纤维化,肾组织羟脯氨酸含量增高,肾功能进行性恶化;而用维生素E治疗的大鼠,氧自由基反应和抗氧化能力趋于正常,肾小球硬化,滤过膜增厚,肾间质纤维化和肾组织羟脯氨酸含量增高的程度显著减轻,肾小球代偿性肥大和肾小球毛细血管代偿性增生的发生后移,肾功能恶化的速度显著减慢。结论:维生素E通过其抗氧化作用,显著抑制5/6肾切除所致慢性肾衰残肾组织纤维化,延缓残肾功能进行性恶化的速度,而对代偿性肾小球肥大和毛细血管增生无明显直接影响。
简介:TodeterminetheregulatoryeffectsofestrogenandcytokineIL-6andIL-8onthegrowthofepithelialovariancancer(OVCA),wefirstexaminedthestatusofestrogenreceptors(ERαandERβ),IL-6receptor(IL-6Rαandgp130),andIL-8receptor(IL-8RAandIL-8RB)onfiveepithelialOVCAcelllinesbysemiquantitativeRT-PCRandWesternblotanalysis.Resultsshowedthattheexpressionsofthesereceptorswerevariableonthefivecells.ThoseOVCAcellsexpressingthereceptorswereselectedtostudyrelatedmolecularmechanism.MTTassaywasperformedtoobservetheeffectsof17β-estradiol(E2),IL-6andIL-8oncellproliferation.WediscoveredthatE2markedlypromotedtheproliferationofCAOV-3andOVCAR-3cellinatime-anddose-dependentmanner.Tamoxifen(Txf),anERinhibitor,completelyblockedtheproliferationoftheE2-inducedcells,andIL-6-or/andIL-8-neutralizingantibodyonlyshowedpartiallyblockingactivity.IL-6andIL-8wereabletosignificantlystimulateCAOV-3andOVCAR-3cellproliferationinatime-anddose-dependentmanner,whichhadapotentialsynergisticeffectonCAOV-3cellsbutnotonOVCAR-3cells.Thecellproliferationinducedbythesetwocytokineswasabolishedcompletelybytheirspecificneutralizingantibodies,partiallybyTxf,butnotbyunrelatedgoatIgG.Takentogether,ourresultssuggestedthatestrogen,IL-6andIL-8couldmodulateOVCAgrowthbyformingareciprocalcascadewithamplifyingeffect.Cellular&MolecularImmunology.