硼替佐米联合顺铂对卵巢上皮性癌顺铂耐药细胞化疗敏感性的影响

Objective To explore the sensitivity and the molecular mechanism of cisplatinresistance ovarian cancer cell line C13 to proteasome inhibitors and the combination with cisplatin. Methods After different treatments, methyl thiazolyl tetrazolium (MTT) assay was applied to examine the cell viability, annexin-V/propidium iodide(PI) apoptosis detection kit was used to determine the apoptosis rate of different groups, western blot assay was introduced to evaluate the expression levels of Fas-associated death domain-like interleukin-1 beta converting enzyme inhibitory protein (cFLIPs), and the activity of caspase-8 was examined. Results MTT assay shown that the cell viability ratios of combination group at serial time points from 12, 24, 36, 48, 60, 72 hours were ( 56.0 ± 8.4 ) %, (44.7 ± 7.3 ) %, ( 33.7 ±11.2) %, (27.6 ± 8.0) %, (27. 6 ± 7.6) % and (28.1 ± 2.4) %, which were much lower than those of cisplatin group (P <0.05). After treated for 24 hours, apoptosis rates of cisplatin group, bortezomib group and combination group were ( 16.7 ± 1.7) %, (23.4 ± 2.1 ) % and (26.9 ± 1.6) %, respectively. The rate of combination group was much higher than that of non-treated group and that of cisplatin group or bortezomib group ( P < 0.05 ). Western blot assay showed the changes of expression levels of cFLIPs, which were downregulated seriously after cisplatin, bortezomib or combination treatment [ (43.2 ± 2.3 )% vs( 75.7 ± 3.0)%vs (67.9 ± 2.1 ) %, P < 0.05 ]. The caspase-8 activity of combination group was (5.6 ± 1.6) folds than that of non-treated group, which was higher than those of other two groups [ ( 2.3 ± 1.0) and (4.2 ± 0.9 ) folds,P < 0.05 ]. Conclusions The tumor cell lethal effect of cisplatin could be increase significantly by the combination application of proteasome inhibitors, bortezomib. And the cFLIPs/caspase-8 signaling pathway may be play an important role in the molecular mechanism of the combination treatment.     

CRF01-AE基因亚型毒株在HIV传播及流行中的作用

目的研究HIV-1CRF01-AE亚型毒株与HIV传播及流行的关系,为预防控制艾滋病提供科学依据。方法分三个时段检测贵州省多地区190份HIV-1毒株样本,用套式PCR扩增其Env基因和测序分析,将实验结果与贵州省艾滋病流行现状进行相关分析。结果检出B,B',E,C亚型及CRF07-BC,CRF08-BC和CRF01-AE三种新的流行性重组亚型毒株,流行性重组亚型毒株是造成贵州省艾滋病流行的因素之一,其中CRF01-AE毒株与性传播、16～50岁年龄段高度相关。结论 CRF01-AE亚型毒株是贵州省艾滋病传播与流行的主要亚型毒株;16～50岁年龄段是CRF01-AE亚型毒株通过性传播艾滋病的主体人群;CRF01-AE亚型毒株引起的艾滋病流行,具有传播范围广、流行时间长、控制难度大等特点。     

中专学校意识形态工作的实践与创新

当前,中等专业学校面临着前所未有的困境,诸如生存与发展的压力、生源紧张、就业难等一系列问题,使这些学校举步维艰.     

中专卫校应加强信息化建设

随着中等医学教育改革的深入，各中专卫校教学条件的改善，以信息化为主体的现代教育技术已逐步向医学教育的各个环节扩展，并成为推动医学教育现代化不可缺少的条件。１加强信息化建设是提高医学生能力素质的需要２１世纪是知识和信息的时代。面对高度信息化的知识和社会生活，获取和处理信息的能力与技巧已成为人类生存的基本素质。《中共中央、国务院关于深化教育改革全面推进素质教育的决定》指出：当今世界，科学技术突飞猛进，知识经济已见端倪，国力竞争日趋激烈。教育在综合国力的形成中处于基础地位，国力的强弱越来越取决于劳动者…     

两种平台法测定HIV感染者CD4计数的比较

[目的]评价全白细胞设门(PanLeucogating,PLG)单平台法对样品采集后不同时间测定的CD4细胞计数的准确性和重复性,以及与传统双平台方法比较之优越性。[方法]采集EDTA抗凝血并置于室温条件下保存。对40份样品用PLG-CD4分析方法进行连续5d的重复测定。对29份样本在采集后d1和d5用PLG-CD4单平台固定法和传统双平台CD4检测法进行平行测定。[结果]PLG-CD4法对新鲜样品分析的准确性与传统的双平台法相当,对陈旧样品分析的准确性明显优于传统的双平台法,PLG-CD4法的准确性及重复性均较好。[结论]PLG-CD4方法简便准确,重复性好;对于陈旧样品的分析有独到的优势。     

广西HIV-1重组流行株env基因V3-V4区序列变异分析

[Objective] To study the genetic variations of V3-V4 and flanking regions of the env gene from the circulating recombinant form （CRF） of human immunodefieiency virus type 1 （HIV-1） in Guangxi. [Methods] 24 samples were collected from the prevailing area of HIV-1 in Guangxi. Fragments of the HIV-1 env gene were amplified by nest-PCR from proviral-DNA templates of HIV-1 infected individuals. The PCR products were then directly sequenced by using ABI 310 DNA SEQUENCER. The sequencesof V3-V4 region and flanking region of the env gene from CRF were carried out amino acid mutations by CLUSTAL. MEGA and dnatools software. [ Results] Of the 24 samples, 21 samples （ 87.5% ） were infected with CRF01-AE. 3 （12.5%） were infected with CRF08-BC. Phylogenetic tree analysis showed that 21 CRF01-AE strains clustered with 97CNGX2F andTHCM240, while 2 CRF08-BC strains were closely related to CRF08-BC.98CN006 and CRF08-BC. 97CNGX6f, and 1 CRF08-BC clustered with C.95IN21068. Nueleotied sequence analysis showed that DNA distances in V3-V4 regions of the env gene of 24 samples were closely related to the normal strains in Guangxi, and majority of amino acid substitution occurred in C3 and V4 regions rather than V3 region. [Conclusions] Majority of genetric variation in V3-V4 regions of the env gene from recombinant form of CRF01-AE occur in the C3 and V4 regions rather than V3 region.     

顺铂诱导宫颈癌Hela细胞凋亡及其作用机制的研究

目的:研究顺铂在体外诱导宫颈癌Hela细胞凋亡及其作用机制。方法:采用MTT法测定顺铂对Hela细胞增殖的影响;流式细胞仪和Hochest33258检测药物作用前后的细胞凋亡情况;RT-PCR检测HPVE6的mRNA水平表达;WesternBlot测定HPVE6、p53、p21、Bax、Bcl-2蛋白水平的表达。结果:顺铂抑制Hela细胞生长呈时效和量效关系;经10μg/ml顺铂分别在12、24、36、48小时作用Hela细胞,亚G1峰与对照组有显著性差异;RT-PCR提示顺铂作用Hela细胞后HPVE6的mRNA水平表达逐渐降低;WesternBlot提示顺铂作用Hela细胞后HPVE6的蛋白水平表达逐渐降低,p53、p21和Bax蛋白水平表达逐步升高,Bcl-2的表达无变化。结论:顺铂通过抑制HPVE6的表达,恢复p53的功能,引起细胞凋亡,起到杀伤肿瘤的作用。     

AKT2和FN蛋白在卵巢癌中的表达及其临床意义

目的 探讨上皮性卵巢癌中蛋白激酶B2(AKT2)及纤维连接蛋白(FN)的表达及其临床意义.方法 采用免疫组织化学SP法及半定量分析方法,检测上皮性卵巢癌(n=50),卵巢良性上皮性肿瘤(n=19)及正常卵巢组织(n=13)中AKT2及FN的表达,并分析AKT2和FN与上皮性卵巢癌病理学分级、临床分期的相关性.结果 AKT2主要表达在细胞质,上皮性卵巢癌中AKT2表达显著高于卵巢良性肿瘤及正常卵巢组织(均P＜0.01);FN主要分布于间质,与正常卵巢组织和良性卵巢肿瘤比较,上皮性卵巢癌组织FN表达明显降低(均P＜0.01);且AKT2、FN表达强度与卵巢癌病理分级及临床分期密切相关(P＜0.01).两者在肿瘤中的表达呈相反的趋势.结论 AKT2表达越高,间质FN表达越低,上皮性卵巢癌浸润程度越高,越容易复发、转移.AKT2、FN的表达可作为临床判断上皮性卵巢癌恶性程度及评估预后有价值的指标.     

湖南省HIV-1耐药警戒线调查

目的 了解湖南省开展艾滋病抗病毒治疗以来,年龄<25岁新近感染HIV-1且未服抗病毒药者出现耐药毒株的变异情况.方法 通过横断面调查,采集15～24岁HIV-1感染者的血清样本.进行HIV-1 pol区耐药基因变异检测分析.结果 69份有效样本中,在RT区1份样本出现可以导致针对NNRTIs系列的NVP、DLV和EFV药物的高度耐药性的K103N、V181C的2个主要耐药突变,1份样本出现可以导致对D4T的低度耐药性和对DDI的潜在耐药性的V75M耐药突变.PR区没有检测到高度耐药突变株.在RT区高度耐药株检出率为2.9%(2/69),说明湖南省HIV耐药株的传播仍处于低度范围内(<5%).结论 表明现有的一线治疗方案是有效的,个体病例尚不需要在治疗开始前进行耐药性检测.但也提示应该加强对早期感染者的耐药监测工作.