miR-200b通过靶向CRKL抑制胶质瘤U251细胞增殖

目的研究hsa-miR-200b对人U251胶质瘤细胞增殖的影响及相关作用机制。方法将miR-200b过表达和表达沉默质粒分别稳定转染至胶质瘤U251细胞,qRT-PCR验证转染效率,CCK-8法检测U251细胞增殖水平变化,qRTPCR以及Western blot方法检测的CRKL的m RNA和蛋白表达水平。双荧光素酶报告基因实验验证miR-200b与CRKL基因3’UTR的结合位点。将CRKL过表达和表达沉默质粒载体分别稳定转染至U251胶质瘤细胞,使用CCK-8法检测U251细胞增殖水平变化。分别建立miR-200b和CRKL的稳定双转染U251细胞系,使用CCK-8法检测对各组细胞增殖水平的变化。结果与对照组相比,miR-200b过表达显著抑制胶质瘤U251细胞的增殖并且抑制CRKL在U251胶质瘤细胞的m RNA和蛋白表达;双荧光素酶报告基因实验验证了miR-200b与CRKL基因3’UTR的结合位点;CRKL过表达显著增强胶质瘤U251细胞增殖能力;CRKL过表达有效阻断了miR-200b抑制胶质瘤U251细胞增殖的作用。结论 miR-200b能够通过靶向下调CRKL抑制胶质瘤U251细胞增殖。     

沉默XIST通过靶向miR-185抑制人胶质瘤U87细胞增殖

目的探讨沉默长链非编码RNA XIST对人胶质瘤U87细胞增殖的影响及可能机制。方法 Lipo3000将XIST沉默质粒载体转染胶质瘤U87细胞后,Real-Time PCR检验转染效率;CCK-8法检测沉默XIST对U87细胞增殖的影响;Real-Time PCR检测转染XIST沉默载体后,U87细胞中miR-185的表达水平变化;双荧光素酶报告基因实验验证XIST与miR-185存在靶向结合;Lipo3000将miR-185过表达和沉默质粒载体分别转染U87细胞,Real-Time PCR检验转染效率;CCK-8法检测转转染miR-185后,U87细胞增殖能力的变化;Lipo3000将miR-185过表达和沉默质粒载体分别转染建立完成的XIST沉默U87细胞系,应用CCK-8法检测U87细胞的增殖水平变化。结果 XIST沉默显著降低了U87细胞的增殖能力,显著提高了miR-185在U87细胞中的表达水平。验证了XIST和miR-185存在着靶向结合。miR-185沉默显著提高了U87细胞增殖能力;显著阻断了沉默XIST下调U87细胞增殖能力的作用;miR-185过表达则作用相反。结论沉默XIST能够通过靶向miR-185,抑制胶质瘤U87细胞增殖。     

miR-429在食管鳞癌中的表达及对细胞增殖迁移的影响

目的探讨微小RNA-429(miR-429)在食管鳞癌(ESCC)中的表达及其对细胞增殖及迁移的影响。方法选取本院45例ESCC患者手术切除的癌组织(ESCC组)及癌旁组织(正常组)标本为研究对象,并采用实时荧光定量(q RT-PCR)检测两组miR-429表达水平。体外培养ESCC癌细胞株系ECa109及人正常食管上皮细胞株Het-1A,将转染试剂、miR-429阴性对照质粒、miR-429mimic质粒分别转染ECa109细胞,命名为空白组、阴性转染组、miR-429过表达组,同时应用生物信息学工具预测miR-429的靶基因为CTl0激酶调节子样蛋白(CRKL),荧光素酶活性检测miR-429与CRKL的靶向调控关系。采用qRT-PCR法检测细胞中miR-429与CRKLmRNA表达水平,蛋白免疫印迹法检测细胞中CRKL蛋白表达。MTT法检测细胞增殖情况,Transwell检测细胞迁移能力。结果与正常组相比,ESCC组miR-429表达水平显著降低(P0.05);荧光素酶活性测定显示3'UTR-Wt细胞中miR-429过表达组荧光素酶活性显著低于阴性对照组(P 0.05),3'UTR-Mut细胞中miR-429阴性转染组与过表达组比较差异无统计学意义(P 0.05);与Het-1A细胞相比,ECa109细胞中miR-429表达水平显著降低(P0.05),而CRKL mRNA及蛋白表达水平显著升高(P 0.05);与空白组、阴性转染组相比,miR-429过表达组miR-429表达水平显著升高(P 0.05),而CRKL mRNA及蛋白表达水平显著降低(P 0.05);与空白组、阴性转染组相比,miR-429过表达组细胞增殖率及迁移数量均显著降低(P 0.05)。结论 ESCC中miR-429呈低表达,其可通过下调CRKL表达进而抑制ESCC细胞增殖及迁移。     

EMAP-Ⅱ上调miR-429表达抑制人U87胶质瘤细胞的增殖、迁移和侵袭能力

目的探讨内皮-单核细胞激活多肽-Ⅱ(EMAP-Ⅱ)对人U87胶质瘤细胞增殖、迁移和侵袭的影响以及可能的机制。方法培养人U87胶质瘤细胞,给予EMAP-Ⅱ处理不同时间后,采用CCK-8检测细胞活力的变化;应用real-time PCR方法检测mi R-429在人U87胶质瘤细胞中的表达变化;利用Lipofect AMINETM2000将antimi R-429及其阴性对照转染至人U87胶质瘤细胞,再给予EMAP-Ⅱ,分别应用CCK-8法和Transwell小室法检测人胶质瘤U87细胞的增殖、迁移及侵袭能力。结果与对照组相比,EMAP-Ⅱ显著抑制了人U87胶质瘤细胞的活力,上调mi R-429在人U87胶质瘤细胞的表达水平,上述变化在EMAP-Ⅱ作用0.5 h时效果最显著;与EMAP-II作用0.5 h组相比,转染anti-mi R-429后,EMAP-Ⅱ的作用被阻断,人胶质瘤U87细胞的细胞活力,以及迁移和侵袭能力基本恢复。结论 mi R-429参与了EMAP-Ⅱ抑制人U87胶质瘤细胞的增殖,迁移和侵袭的调控。     

过表达miR-139-5p对胶质母细胞瘤U87MG细胞增殖、侵袭的影响

目的 探讨过表达miR-139-5p对胶质瘤U87MG细胞增殖和侵袭的影响,并初步分析其可能机制.方法 在胶质瘤U87MG细胞中瞬时转染miR-139-5p并用real-time PCR方法验证其转染效率,CCK-8法检测转染后各组U87MG细胞的增殖能力,Transwell实验检测转染后各组U87MG细胞的侵袭能力,Western blot检测转染后各组U87MG细胞中BMI1蛋白的表达,应用生物信息网站Targetscan和双荧光素酶报告基因实验验证BMI1是否为miR-139-5p的靶基因.结果 miR-139-5p过表达使U87MG细胞增殖能力下降,侵袭能力下降,U87MG细胞中BMI1蛋白表达下降,生物信息学分析及双荧光素酶实验证实BMI1为miR-139-5p的靶基因.结论 过表达miR-139-5p抑制U87MG细胞增殖及侵袭能力,与下调BMI1的表达相关.     

探讨人胶质瘤细胞系U87细胞中反义miR-21诱导凋亡机制

目的 探讨下调miR-21诱导人胶质瘤细胞系U87细胞凋亡机制.方法 应用化学方法合成的反义miR-21寡聚核苷酸(anti-miR-21),通过瞬转法转染U87细胞,检测U87细胞凋亡、增殖、侵袭等变化,并结合生物信息学分析、Western印迹验证在U87细胞中miR-21和PTEN基因及caspase间关系.结果 体外转染反义miR-21寡聚核苷酸能明显抑制U87细胞生长,诱导其凋亡,降低侵袭能力;增加caspase-3表达活性,活化caspase-9表达,但不影响PTEN和caspase-8表达.结论 miR-21可能是人胶质瘤U87细胞的抗凋亡微RNA(microRNA,miRNA),反义miR-21可能通过easpase-9、3而不是PTEN诱导肿瘤细胞凋亡.     

miR-137对喉癌Hep-2细胞增殖、侵袭与凋亡的影响及可能机制

目的探讨miR-137对喉癌Hep-2细胞增殖及迁移能力的影响及可能机制。方法采用miR-137模拟物转染喉癌Hep-2细胞,实验分为空白对照组组(未进行转染)、miR-137阴性对照组(转染无关序列)、miR-137模拟物转染组(进行miR-137模拟物转染)。real-time PCR法验证转染效率,采用CCK8法检测miR-137对喉癌Hep-2细胞增殖的影响,采用Transwell实验检测转染后Hep-2细胞的侵袭能力变化,采用流式细胞术检测miR-137对Hep-2细胞凋亡的影响;Western blot检测转染后Hep-2细胞凋亡相关蛋白Bcl-2、Bax、caspase-3以及侵袭相关蛋白T细胞因子-4(TCF-4)蛋白表达的变化。结果与空白对照组或阴性对照组相比,miR-137模拟物转染组Hep-2细胞miR-137的表达水平显著高于对照组(P0.01),喉癌Hep-2细胞的增殖与侵袭能力显著降低,细胞凋亡率明显升高,凋亡相关蛋白Bcl-2表达水平降低,而Bax与caspase-3蛋白表达升高,侵袭相关蛋白TCF-4表达降低。结论过表达miR-137抑制Hep-2细胞增殖与侵袭、诱导细胞凋亡,与上调Bax、caspase-3蛋白表达和下调Bcl-2、TCF-4表达相关。     

miR-342对胶质瘤细胞增殖,迁移侵袭及凋亡的影响

目的研究miR-342的表达变化对胶质瘤细胞生物学行为的影响。方法应用real-time PCR方法检测miR-342的内源性表达。CCK-8检测细胞增殖,Transwell法检测细胞的迁移侵袭,流式细胞仪检测细胞的凋亡。结果miR-342在胶质瘤细胞中表达水平较正常脑组织低;过表达miR-342能够抑制U87胶质瘤细胞的增殖、迁移侵袭,促进凋亡;相反,表达沉默miR-342能够促进U251胶质瘤细胞增殖、迁移侵袭,并抑制细胞凋亡(P0.05)。结论miR-342能够调控胶质瘤细胞的增殖、迁移侵袭,起抑癌基因的作用。     

MACC1表达沉默对胶质瘤细胞U87增殖凋亡的影响

目的探讨MACC1基因表达沉默对脑胶质瘤细胞U87增殖凋亡的影响。方法应用RT-PCR和Western blot检测55例不同病理级别胶质瘤标本MACC1表达;转染MACC1siRNA到U87细胞沉默MACC1基因表达,Western blot检测MACC1,Bcl-2/Bax,caspase-3,c-Met蛋白的表达;MTT法检测细胞生长增殖能力;流式细胞术检测细胞周期及凋亡情况。结果MACC1基因在胶质瘤标本组织表达随病理级别升高而增多;MACC1表达沉默后U87生长增殖能力明显下降,细胞周期阻滞于G0/G1期,细胞凋亡显著增加,MACC1蛋白表达明显下降,Bcl-2/Bax比值降低,caspase-3表达上调,c-Met表达下降。结论下调MACC1表达可促进胶质瘤细胞凋亡,抑制肿瘤细胞增殖,MACC1可能成为胶质瘤基因治疗的新靶点。     

miR-379对胶质瘤U87MG细胞增殖、迁移和侵袭能力的影响

目的研究miR-379对胶质瘤U87MG细胞增殖、迁移和侵袭能力的影响。方法 Real-time PCR方法检测miR-379在胶质瘤U87MG细胞和正常星型胶质细胞中的表达水平。在胶质瘤U87MG细胞中瞬时转染miR-379 agomir并用real-time PCR方法验证其转染效率。CCK-8方法检测miR-379对胶质瘤U87MG细胞增殖能力的影响。划痕实验检测miR-379对胶质瘤U87MG细胞迁移能力的影响。Transwell侵袭实验检测miR-379对胶质瘤U87MG细胞侵袭能力的影响。结果 miR-379在胶质瘤U87MG细胞中的表达水平显著低于在正常星型胶质细胞中的表达。miR-379 agomir抑制胶质瘤U87MG细胞的增殖、迁移和侵袭能力。结论 miR-379抑制胶质瘤U87MG细胞的增殖、迁移和侵袭能力。     

Level of functioning of siblings and parents of probands of varying degrees of retardation

A further analysis of already published data supports the position that retardates of low ability level less frequently have retarded siblings, retarded parents, and parents low in occupational level than do retardates higher in ability level. The analysis supports the position that there are two types of retarded individuals, persons retarded as a result of gene or chromosomal anomalies, brain injury, etc., who more frequently occur in the lower-level retardate group, and persons whose retardation represents polygenic segregation, who more frequently occur in the higher-level group.     

Modes of Inheritance of Errors of Refraction

Eighteen families in which both parents had refractions within the range of +4·0 D to −4·0 D and axial lengths seen in emmetropia (22·3-26·0 mm) showed coefficients of correlation of the order 0·5 indicative of polygenic inheritance. Such coefficients were seen for axial length (0·407) and for the cornea (0·487), but not for the lens (which is known to be yoked to the axial length). No such coefficients were seen in 19 families in which one of the parents had axial length outside the emmetropic range (nine families with long axes and 10 with short axes).

The pattern of polygenic inheritance for emmetropia (completely correlated optical components) and errors of refraction up to 4·0 D (inadequately correlated components: correlation ametropia) follows that seen in stature and other measurable characters. In contrast the high refractive errors with their abnormal axial lengths (component ametropia) are—like the extremes in stature—pathological anomalies with monofactorial inheritance.

     

Aspects of the problem of direct introduction of Standards of International Organizations

Editorial note. This article is published as part of a discussion. Particular issues of the article are disputable. First of all, this concerns the so-called “folder” method of introduction of international standards for medical devices to domestic medical practice (i.e., by direct translation of the standards and their publication as standardizing documents). Nevertheless, at least one of the problems, the problem of coordination between domestic state standards for medical devices and international recommendations of ISO and IEC, is undoubtedly of topical importance. Advancement of new health service legislation which is to be approved by law-makers will definitely introduce corrections into the present situation. The Editorial Board of Meditsinskaya Tekhnika believes this article will lessen these problems and to be welcomed by readers.