miR-195-5p表达上调抑制肺癌细胞系A549的迁移和侵袭

目的探讨miR-195-5p对肺癌细胞系A549细胞迁移和侵袭的影响及其机制。方法实时荧光定量PCR检测肺癌组织及肺癌细胞系中miR-195-5p的表达;通过脂质体介导将miR-195-5p模拟物作用于A549细胞,用Transwell法检测细胞迁移和侵袭; Western blot检测细胞内上皮间质转化标志物及ZEB1的表达。结果相对于癌旁组织及人正常肺上皮细胞,miR-195-5p在肺癌组织及肺癌细胞系中低表达(P0. 001);过表达miR-195-5p可显著抑制A549细胞的迁移和侵袭(P0. 001),伴随E-cadherin蛋白表达的上调,N-cadherin和vimentin蛋白表达的下调(P0. 01);过表达miR-195-5p可抑制A549细胞ZEB1的表达(P0. 001)。结论 miR-195-5p可能通过下调ZEB1的表达,抑制肺癌细胞迁移和侵袭。     

miR-125a-5p靶向LIMK1逆转非小细胞肺癌A549/DDP细胞对顺铂的耐药性

目的:探讨微小RNA-125a-5p(miR-125a-5p)对非小细胞肺癌A549/DDP细胞对顺铂耐药性的影响及其作用机制。方法:RT-qPCR检测非小细胞肺癌组织及其细胞株A549和A549/DDP中miR-125a-5p和LIM激酶1(LIMK1)的表达;在A549/DDP细胞中上调或下调miR-125a-5p或LIMK1表达,分别采用MTT法、流式细胞术和Western blot检测细胞活力、凋亡率及耐药相关蛋白表达;TargetScan在线预测、萤光素酶报告基因实验验证miR-125a-5p和LIMK1的靶向关系;共表达miR-125a-5p和LIMK1,检测细胞活力、凋亡率及耐药相关蛋白表达。结果:在非小细胞肺癌组织及其细胞株中,miR-125a-5p表达下调,LIMK1表达上调(P0.05);萤光素酶报告基因实验表明miR-125a-5p可负向调控LIMK1表达。过表达miR-125a-5p或敲减LIMK1表达使A549/DDP细胞的活力下降,细胞凋亡率增加,顺铂的IC_(50)减小,耐药相关蛋白表达下调(P0.05)。过表达LIMK1则使miR-125a-5p对A549/DDP细胞活力和耐药相关蛋白表达的抑制作用减弱。结论:miR-125a-5p能通过抑制LIMK1基因表达,下调耐药相关蛋白表达,从而逆转A549/DDP细胞对顺铂的耐药性。     

circDCUN1D4调节miR-18a-5p/FBP1轴对肺癌细胞增殖、凋亡和免疫逃逸的影响

目的 探讨环状RNA(circRNA)DCUN1D4调节微小RNA(miR)-18a-5p/果糖-1,6-二磷酸酶1(FBP1)轴对肺癌细胞增殖、凋亡和免疫逃逸的影响。方法 选取人肺癌细胞系（H1975、H1650、A549、SPCA-1）和人正常肺表皮细胞（HPL-1）,qRT-PCR检测各种细胞中circDCUN1D4、miR-18a-5p、FBP1 mRNA表达水平。取对数生长期的A549细胞并分为空白组、circDCUN1D4过表达质粒（circDCUN1D4）组、过表达质粒阴性对照（NC）组、circDCUN1D4+miR-18a-5p模拟物阴性对照（circDCUN1D4+mimics NC）组、circDCUN1D4+miR-18a-5p模拟物（circDCUN1D4+miR-18a-5p mimics）组。CCK-8法检测各组A549细胞活力,流式细胞术检测各组A549细胞凋亡水平,qRT-PCR检测各组A549细胞中circDCUN1D4、miR-18a-5p、FBP1 mRNA表达水平,Western blot检测各组A549细胞中FBP1、caspase-3、Ki...     

LncRNA SLC16A1-AS1 通过靶向 miR-15a-5p 负向调控子宫颈癌细胞的增殖和侵袭

目的探讨LncRNA SLC16A1-AS1通过靶向miR-15a-5p调控子宫颈癌细胞增殖和侵袭的作用机制。方法采用qRT-PCR法检测SLC16A1-AS1在子宫颈癌组织、癌旁组织、人正常子宫颈上皮细胞(H8)及子宫颈癌细胞株(HeLa、HCC94、SiHa、C33A)中的表达水平。选择SLC16A1-AS1表达最少的子宫颈癌细胞株,分别转染阴性质粒和SLC16A1-AS1过表达质粒,标记为NC组和SLC16A1-AS1组。应用MTT法和Transwell实验分别检测过表达SLC16A1-AS1对子宫颈癌细胞增殖和侵袭能力的影响。生物信息学网站预测及双荧光素酶报告基因实验,验证SLC16A1-AS1与靶基因的靶向关系。qRT-PCR和Western blot法检测靶基因及相关蛋白的表达。结果子宫颈癌组织中SLC16A1-AS1的表达水平显著低于癌旁组织(P0.01)。与H8细胞相比,HeLa、HCC94、SiHa、C33A细胞中SLC16A1-AS1的表达水平降低(P0.05)。与NC组相比,过表达SLC16A1-AS1的C33A细胞增殖能力降低(P0.05),过表达SLC16A1-AS1的C33A细胞侵袭数下降(P0.01)。生物信息学网站预测显示,SLC16A1-AS1可与miR-15a-5p有特异性结合位点。双荧光素酶报告基因实验结果证实SLC16A1-AS1可与miR-15a-5p直接结合(P0.01)。SLC16A1-AS1可负向调控miR-15a-5p的表达(P0.01)。与NC组相比,SLC16A1-AS1组细胞增殖表型蛋白(如PCNA、Ki-67)和细胞侵袭表型蛋白(如N-cadherin、Slug)表达均降低。结论 SLC16A1-AS1在子宫颈癌中低表达,SLC16A1-AS1通过靶向下调miR-15a-5p的表达,抑制子宫颈癌细胞C33A的增殖和侵袭。     

miR-708-5p通过靶向URGCP抑制非小细胞肺癌A549细胞侵袭、转移的研究

目的 探讨miR-708-5p对非小细胞肺癌A549细胞的增殖、迁移和侵袭的影响及其与上调基因-4(upregulated gene-4/upregulator of cell proliferation,URG4/URGCP)的靶向关系.方法 选取非小细胞肺癌细胞株A549,正常肺成纤维细胞HLF-1为研究对象,根据A549细胞转染物质的不同,分为Control组(未进行任何处理),NC组(转染随机序列RNA Oligo),miR-708-5p组(转染miR-708-5p mimics).MTT检测增殖能力,流式细胞术检测细胞周期,Transwell小室实验检测侵袭能力,划痕实验检测迁移能力,Western blot检测URGCP表达水平;实时荧光定量PCR检测miR-708-5p、URGCP基因mRNA表达水平;荧光素酶实验验证miR-708-5p与URGCP的靶向关系.结果 与HLF-1细胞株相比,肺癌细胞株A549细胞miR-708-5p表达水平降低,URGCP-mRNA表达升高(P<0.05),同时,URGCP蛋白水平升高.转染72h后,与Control、NC组比较,miR-708-5p组细胞URGCP基因mRNA明显降低,URGCP蛋白表达量明显降低(P均<0.01).miR-708-5p组细胞增殖、迁移和侵袭能力明显低于Control、NC组(P<0.05).与WT-URGCP组相比,miR-708-5p+WT-URGCP组细胞荧光素酶活性明显降低(P<0.01).结论 miR-708-5p在非小细胞肺癌A549细胞中低表达,过表达miR-708-5p可抑制A549细胞增殖、迁移和侵袭,机制可能与负性调控URGCP有关.     

miR-655-3p靶向KIF20A抑制人肺腺癌细胞系A549迁移及侵袭

目的 探讨miR-655-3p靶向驱动蛋白家族成员20A(KIF20A)对肺腺癌细胞增殖、迁移及侵袭的影响.方法 RT-qPCR检测人肺腺癌细胞系H460、A549、HCC-2935、H1299细胞中miR-655-3p的表达;取对数增殖期的A549细胞,分为:空白组、miR-NC组、miR-655-3p mimics...     

miR-148a-3p靶向DNMT1表达调控肺癌细胞A549增殖、迁移和侵袭作用研究

目的 探讨miR-148a-3p靶向DNMT1表达调控肺癌细胞A549增殖、迁移和侵袭的机制.方法 设肺癌细胞A549组、miR-NC组、miR-148-3p mimics组、miR-148-3p inhibitor组,以上各组细胞每孔设6个平行样,培养72 h.采用MTT法检测细胞活力,甲基紫染色测定细胞单克隆形成数...     

circAGFG1对非小细胞肺癌生物学行为的影响机制研究

目的 探讨环状RNA(circRNA)AGFG1通过靶向微小RNA(miR)-374a-5p/血管内皮生长因子C(VEGFC)轴对非小细胞肺癌（NSCLC）生物学行为的影响机制。方法 qRT-PCR检测55例NSCLC患者的癌组织和癌旁组织以及人支气管上皮细胞系（BEAS-2B）、人NSCLC细胞系（HCC827、H1975、A549、H1299）中circAGFG1、miR-374a-5p、VEGFC mRNA表达水平。以A549细胞为研究对象,设置对照组、circAGFG1小干扰RNA(si circAGFG1)组及阴性对照（si NC）组、si circAGFG1+miR-374a-5p抑制物（miR-374a-5p inhibitor）组以及si circAGFG1+抑制物对照组（inhibitor NC）组。qRT-PCR检测各组A549细胞中circAGFG1、miR-374a-5p、VEGFC mRNA表达水平;CCK-8及Transwell实验分别检测细胞活力及迁移、侵袭能力;Western blot检测增殖蛋白Ki-67、基质金属蛋白酶-9(MMP-9)以及VEGFC...     

miR-101及Runt相关转录因子1在肺癌细胞系A549中的作用

目的研究miR-101及其下游靶基因Runt相关转录因子1(RUNX1)在肺癌细胞系A549中的作用。方法 Western Blot及Real Time PCR检测miR-101和RUNX1在A549中的表达情况,上调或下调miR-101后A549中RUNX1的表达变化,绘制生长曲线及细胞克隆实验检测A549细胞的增殖变化,transwell验证A549细胞迁移的变化。结果在A549细胞,miR-101表达水平下调,RUNX1表达水平上调;miR-101抑制A549细胞的增殖及迁移;RUNX1促进A549细胞的增殖及迁移。结论 miR-101通过抑制RUNX1的转录表达抑制肺癌细胞系A549的增殖及迁移。     

miR-409-3p靶向癌基因ELF2抑制非小细胞肺癌细胞系A549增殖

目的观察miR-409-3p对非小细胞肺癌细胞系A549增殖的影响及其机制。方法对照组为常规条件下培养的转染阳性对照核苷酸的A549细胞寡核苷酸组。实验组在对照组的基础上采用化学合成miR-409-3p模拟物,脂质体转染构建miR-409-3p高表达A549细胞;经q PCR检测,观察2组48 h后miR-409-3p的表达; MTT法检测细胞增殖;流式细胞计量术检测细胞凋亡和周期的差异。结果 miR-409-3p模拟物转染后,A549细胞中miR-409-3p表达水平显著升高(P0. 05);细胞总凋亡比例为22. 66%±4. 61%,显著高于对照组的7. 78%±1. 95%(P0. 05); G0/G1期为40. 21%±5. 37%,显著低于对照组细胞56. 09%±5. 22%(P0. 05);双荧光报告系统表明癌基因ELF2是miR-409-3p的靶基因,过表达miR-409-3p模拟物后,内源性ELF2蛋白表达水平显著下降(P0. 05)。结论 miR-409-3p可能通过调控ELF2的转录水平抑制非小细胞肺癌细胞A549增殖并促进其凋亡。     

某幼儿园一起手足口病暴发的现场流行病学调查报告

目的了解某幼儿园手足口病暴发的流行病学特征及造成传染的主要原因,为进一步做好该园手足口病防控工作提供依据。方法按照病例定义,电话开展病例搜索,结合现场流行病学调查情况,形成假设,采用1：2匹配的病例对照研究,对照选取同班同年龄组同性别的儿童,使用统一的调查表询问儿童的日常卫生习惯。结果该幼儿园从2010年3月16日至3月23日共发生9例临床诊断病例,3例实验室确诊病例,罹患病率为7.95%。以小班病例为主,3～4岁组比例高（8/12）。3份粪便检测出病原体CoxA16。有吸吮手指、吸吮玩具、饭前不洗手、便后不洗手、挖鼻孔、揉眼睛等任一不良卫生习惯是手足口病发病的危险因素（OR=12,95%CI=1.3～114）。结论幼儿不良卫生习惯是该幼儿园手足口病传播的主要原因。建议该园加强儿童不良卫生习惯的纠正,培养良好卫生习惯,以达到控制传染的目的。     

Characteristics of transducing group A streptococcal bacteriophages A 5 and A 25

Summary The virulent, transducing, DNA-containing group A streptococcal bacteriophages A5 and A25 are 58 m in head size and have a flexible, noncontractile tail with a length of 180 m. The morphologic outline of the head is hexagonal. Treatment of the bacteriophages with potentially lethal ultraviolet radiation produced damages which were repairable by the Hcr⁺ streptococcal host strain K56, the host-cell reactivable sector being 0.9. These properties together with a demonstrable serological cross reaction show a close relationship between the two phage strains.     

A case of prenatal diagnosis of hemophilia A.

Classic hemophilia, (hemophilia A), is an X-linked hereditary bleeding disorder affecting half of the male offspring of female carriers. Prenatal diagnosis offers an option, namely to restrict abortions to hemophilic fetuses only, and thus retain the chance of bearing normal sons. Recently, the authors have made a prenatal diagnosis of hemophilia A in an obligate carrier with a male fetus at 24 weeks of gestation by pure fetal sampling and accurate factor VIII coagulant assay, which was repeatedly less than 1% at 28 weeks of gestation.     

A different view of A68 and amyloid

Data discussed in Dr. Selkoe's review can be interpreted in several ways, suggesting alternate hypotheses regarding the nature of A68, and the relationship between amyloid and Alzheimer's disease.     

A rare form of spondylometaphyseal dysplasia-type A4

We present 2 cases of a previously apparently unreported spondylo-metaphyseal dysplasia comprising dwarfism, severe metaphyseal changes, ovoid vertebrae and mild platyspondyly with anterior tonguing of the vertebral bodies. The inheritance may be autosomal recessive. Am. J. Med. Genet. 78:61–66, 1998. © 1998 Wiley-Liss, Inc.     

Phospholipase A — A Parameter of Sepsis?

Summary Phospholipase A (PLA) and Sepsis Severity Score (SSS) were measured regularly in 28 patients with sepsis (n=11), pancreatic operations (n=7), or multitrauma combined with contused abdominal trauma (n=10). No linear correlation was found between these two parameters. A statistical correlation could be shown for the paired values PLA_max/SSS or PLA/SSS_max. They lie together above or below their critical value of 20 points SSS or 30 U/l PLA (<0.05). The evaluation of mortality shows a distinctly higher significance for the SSS (P<0.01) in comparison with the PLA (P<0.05). In certain cases PLA was the first parameter which could have shown the beginning of a septic process.     

A new continuous epitope of hepatitis A virus

A new continuous epitope of hepatitis A virus (HAV) was defined in the VP3 protein. Convalescent sera recognised the synthetic peptide 3110–3121 (FWRGDLVFDFQV). The replacement of the arginine, glycine, or aspartic acid at positions 112, 113, or 114, respectively by other aminoacids induced the loss of synthetic peptide recognition by human convalescent sera, thereby confirming the presence of an epitope in the original VP3(110-121) sequence. Shorter VP3 peptides such as VP3(110-119), VP3(110-117), and VP3(110-116) and a tandem repeat of VP3(111-116) failed to react with convalescent sera, indicating the importance of the entire peptide in the epitope structure. The maximum inhibition of human convalescent binding to HAV by the VP3(110-121) peptide was around 60%, and 50% inhibition was achieved at a peptide concentration of 2.3–2.4 μg/ml. Antibodies generated by this peptide bound to intact HAV and neutralised its infectivity. Antipeptide antibodies inhibited convalescent serum binding to HAV. Monoclonal antibodies H7C27 and MAK-4E7 inhibited completely binding of the antipeptide antibodies to HAV. J. Med. Virol. 54:95–102, 1998. © 1998 Wiley-Liss,Inc.