外源性硫氧还蛋白对高糖、高脂刺激引起的H9c2细胞损伤和凋亡的影响

目的 观察给予外源性硫氧还蛋白（Trx）后对高糖、高脂刺激引起的心肌细胞损伤和凋亡的影响,并分析其可能机制.方法 将处于对数生长期的H9c2心肌细胞随机分为三组：正常对照组（普通DMEM培养基葡萄糖5 mmol/L加入20 mmol/L甘露醇）、高糖高脂组（DMEM高糖25 mmol/L培养基加入高脂600 μmol/L软脂酸）和Trx干预组（在高糖高脂组的培养基中加入3 ＊9滋mol/L Trx）.培养48 h后,分别收集培养基上清与细胞,进行指标检测.结果 与正常对照组比较,高糖高脂组乳酸脱氢酶及caspase-3活性均显著升高（P＜0.01）.给予Trx干预后,与高糖高脂组比较,Trx干预组乳酸脱氢酶及caspase-3活性均显著下降（P＜0.01）.与正常组比较,高糖高脂组Trx的表达量未见明显改变（P＞0.05）,但是Trx活性显著下降（P＜0.01）.结论 高糖、高脂刺激可以引起H9c2心肌细胞损伤和凋亡,其机制与内源性Trx活性的降低有关.外源性补充Trx可明显提高Trx的活性,减轻心肌细胞的损伤和凋亡.     

Seroprevalence of pandemic (H1N1) 2009 influenza and effectiveness of 2010/2011 influenza vaccine during 2010/2011 season in Beijing, China

Please cite this paper as: Yang et al. (2011) Seroprevalence of pandemic (H1N1) 2009 influenza and effectiveness of 2010/2011 influenza vaccine during 2010/2011 season in Beijing, China. Influenza and Other Respiratory Viruses 6(6), 381–388. Background In the post‐pandemic period, pandemic (H1N1) 2009 virus was expected to circulate seasonally and was introduced into trivalent influenza vaccine during 2010/2011 season in the Northern Hemisphere. Objectives The aim of this study was to examine the evolution of herd immunity against pandemic (H1N1) 2009 virus in Beijing, China, during 2010/2011 season and effectiveness of the 2010/2011 trivalent vaccine. Methods Two serological surveys were conducted before and after 2010/2011 season in Beijing. A case–control study was used to investigate vaccine effectiveness against influenza‐like illness (ILI) and lower respiratory tract infection (LRI). Results A total of 4509 and 4543 subjects participated in the pre‐ and post‐season surveys, respectively. The standardized seroprevalence of pandemic (H1N1) 2009 influenza increased from 22·1% pre‐season to 24·3% post‐season (P < 0·001). Significant elevation in seroprevalence appeared in the ≥60 years age‐group (P < 0·001), but not in others. The 2010/2011 trivalent vaccine contributed to the higher post‐seasonal seroprevalence in unvaccinated individuals (P = 0·024), but not in those vaccinated with monovalent pandemic vaccine (P = 0·205), as well as in those without prior immunity versus those with immunity. The adjusted effectiveness of the 2010/2011 trivalent vaccine was 79% protection against ILI (95% CI, 61–89%) and 95% against LRI (95% CI: 59–99%). Conclusions A slight increase in herd immunity against pandemic (H1N1) 2009 influenza was observed in Beijing, China, during the 2010/2011 season. Prior vaccination and immunity had a suppressive impact on immune response toward this novel influenza virus, elicited by 2010/2011 trivalent vaccine. This trivalent vaccine conferred good protection against ILI and LRI.     

RNA干扰抑制EZH2基因表达对人胶质瘤细胞凋亡及对卡莫司汀敏感性的影响

目的观察RNA干扰技术抑制EZH2基因表达对人胶质瘤U251细胞凋亡及其对卡莫司汀敏感性的影响。方法针对EZH2 mRNA序列设计合成小干扰RNA（siRNA）的DNA模板,构建pRNAT-EZH2重组表达载体,并用其转染人胶质瘤U251细胞。用RT-PCR法检测pRNAT-EZH2转染前后U251细胞中内源性EZH2表达;激光共聚焦显微镜观察转染后及加入卡莫司汀后U251细胞凋亡情况;MTT法检测转染前后U251细胞对化疗药物卡莫司汀敏感性;用分光光度法检测转染前后以及加入卡莫司汀后U251细胞中Caspase-3活性。结果成功构建pRNAT-EZH2重组质粒,并成功转染U251细胞。pRNAT-EZH2重组质粒转染后U251细胞中EZH2 mRNA表达量降低（P〈0.01）;卡莫司汀组及卡莫司汀加pRNAT-EZH2组U251细胞凋亡明显;与卡莫司汀联合时,pRNAT-EZH2组U251细胞生长抑制率明显增高（P〈0.01）;Caspase-3活性明显高于空白对照组和阴性对照组（P均〈0.01）。结论pRNAT-EZH2可抑制EZH2在人胶质瘤U251细胞中的表达,并增强U251细胞对卡莫司汀敏感性。     

A plea for caution: huge risks associated with lab-bred flu

I wish to express concern about the maintenance of laboratory strains of H5N1 influenza viruses that might be adapted for transmission among humans.     

Fimbriation and curliation in Escherichia coli O157:H7: a paradigm of intestinal and environmental colonization

Shiga toxin-producing Escherichia coli (STEC) serotypes, particularly E. coli O157:H7, possess a variety of fimbrial and afimbrial adhesins which have emerged as important contributors to intestinal colonization. E. coli O157:H7 possesses two chromosomal operons encoding long polar fimbriae (Lpf), which have been found to influence adherence in vitro and colonization in vivo. In a recent Infection and Immunity paper, we further explored the role of Lpf in E. coli O157:H7 intestinal colonization by using the infant rabbit model of STEC infection. We found that an E. coli O157:H7 Lpf-deficient mutant was outcompeted in the rabbit intestine by its parental strain, which may suggest that Lpf contributes to colonization. In contrast, the Lpf-deficient mutant showed an increased adherence to cultured intestinal epithelial cells, and we discovered that this strain overexpressed curli fibers. In this addendum article, we provide a continued perspective on the predicted roles of Lpf and curli, both in vivo and in vitro.     

exosomes在肺癌细胞对顺铂敏感性降低过程中的作用研究

目的研究肺癌细胞分泌的exosomes在同源细胞对顺铂的敏感性调节过程中的可能作用。方法采用超速离心和Exoquick-TC结合的方法从肺癌细胞系A549和H1975的上清液中分离出exosomes,透射电子显微镜观察其形态,western blot检测其CD63蛋白表达;共聚焦显微镜观察细胞胞吞exosomes过程;分别提取顺铂处理肺癌细胞来源exosomes和未处理肺癌细胞来源exosomes,并使用2种exosomes分别处理肺癌细胞,采用CCK-8法检测上述获得的exosomes对肺癌细胞顺铂敏感性的影响。结果透射电子显微镜下观察肺腺癌细胞来源的exosomes具有特征性的盘状结构,直径30-100 nm,western blot结果显示exosomes富含CD63蛋白;显微镜下可观察到exosomes可进入细胞;同时经顺铂处理过肺癌细胞分泌的exosomes可降低同源细胞对顺铂的敏感性。结论减少exosomes的分泌和传递可能会增加顺铂化疗的疗效,这为肺癌的治疗提供了一种新的思路。     

十堰地区妇女宫颈人乳头瘤病毒感染及基因亚型分布情况调查分析

目的:对十堰地区妇女宫颈(HPV)亚型进行筛查,以探讨其分布规律。方法:采用核酸分子快速导流杂交基因芯片技术,对1 116例妇科就诊患者的宫颈刷片标本进行了21种HPV基因分型检测。结果:1 116例样本中,HPV感染者283例,总感染率25.36%,高危型HPV感染率14.52%,低危型HPV感染率2.96%,高危型及低危型混合感染阳性率7.89%;单一基因型别195例,占阳性患者的68.91%;双重混合感染59例,占阳性患者的20.85%;三重以上混合感染者29例,占阳性患者的10.25%。HPV感染阳性率居前5位的高危亚型依次是HPV16型50例(4.48%)、HPV52型47例(4.21%)、HPV53型28例(2.51%)、HPV58型17例(1.52%)及HPV39型15例(1.34%),常见的低危型是HPV6(1.26%)和HPV42(1.26%)。结论:十堰地区妇女HPV感染率较高,且以高危型HPV感染为主,感染型别既符合亚洲及中国地区人群的分布规律,又有独特的区域分布特点,并且多重感染率高,值得重视。     

肠出血性大肠埃希菌O157∶H7环介导恒温扩增快速检测方法的建立与应用

目的肠出血性大肠埃希菌(Enterohemorrhagic Escherichia coli,EHEC)O157∶H7是一种重要的人畜共患病致病菌,主要通过被污染的食物传播,引起出血性结肠炎,建立其快速检测方法具有重要意义。方法基于EHEC O157∶H7保守的rfbE基因序列,设计4条引物,利用环介导等温扩增技术(loop-mediatedisothermal amplification,LAMP),成功建立了EHEC O157∶H7LAMP快速检测方法 ,60min内即可完成致病菌检测。结果利用该LAMP方法对30种共38株细菌进行检测,所试EHEC O157∶H7均为阳性结果 ,说明该方法具有高度特异性。本方法对纯培养的EHEC O157∶H7检测限为12CFU/mL,污染食品中EHEC O157∶H7的检测限为18CFU/g。实践应用表明,对1121份进出口肉类、奶类制品及人工污染样品等进行检测,检出57份LAMP阳性,与采用AOAC标准检测结果的符合率为100%。结论该LAMP方法操作简便、特异性强、灵敏度高,具有良好的实用性。     

含H5N1禽流感病毒核酸序列的病毒样颗粒的制备及应用

目的构建耐RNase酶的内含禽流感病毒H5N1亚型部分核酸序列的病毒样颗粒。方法构建中间载体pET32a-MS2,将禽流感病毒H5N1亚型的HA、NA和M基因片断连接到中间载体上,构建原核表达载体pET32a-MS2-HA,pET32a-MS2-NA和pET32a-MS2-M,分别转化宿主菌,诱导表达制备病毒样颗粒。结果表达载体经PCR、酶切鉴定和测序分析后证实构建成功。电镜观察到了病毒样颗粒,荧光定量RT-PCR试验表明颗粒稳定性良好。结论成功构建了含禽流感病毒H5N1部分核酸序列的病毒样颗粒且稳定性良好,可作为禽流感H5N1亚型RNA检测的标准品和质控品。