葡萄糖调节蛋白78真核表达载体及稳定转染人视网膜色素上皮细胞株的构建

背景：葡萄糖调节蛋白78是存在于内质网上最多的分子伴侣蛋白,具有保护细胞对抗细胞调亡的作用。 目的：构建携带并正确表达外源性人葡萄糖调节蛋白78基因的重组真核表达载体,建立葡萄糖调节蛋白78基因稳定转染的人视网膜色素上皮细胞系。 方法：扩增人葡萄糖调节蛋白78全长编码序列,将该目的基因与真核表达载体PEGFP-N1进行定向连接,其产物转化细菌感受态细胞,以PCR方法鉴定阳性克隆,并进行测序和分析比对,获得融合蛋白表达质粒载体pEGFP-grp78。通过阳离子脂质体介导将重组质粒pEGFP-grp78转染入体外培养的人视网膜色素上皮细胞,经G418筛选,建立稳定表达细胞系。 结果与结论：体外培养的人视网膜色素上皮细胞株作为真核细胞表达系统,经荧光显微镜和RT-PCR方法检测,G418筛选的稳定转染细胞系中GFP大量表达,葡萄糖调节蛋白78 mRNA的表达量是正常视网膜色素上皮细胞的4倍左右,表示脂质体介导的pEGFP-grp78质粒成功转染视网膜色素上皮细胞,并高效稳定表达葡萄糖调节蛋白78。     

SDZ对大肠癌lovo细胞的影响

目的 观察SDZ体外对大肠癌lovo细胞的影响.方法 培养大肠癌lovo细胞,加入不同浓度的SDZ,作用24 h、48 h、72 h,用MTT法计算细胞生长抑制率,描绘细胞生长曲线.结果 SDZ体外对大肠癌lovo细胞的生长抑制率随作用浓度和作用时间的增加而增大.1 000μg/mL,200μg/mL,40 μg/mL,8 μg/mL,1.6μg/mL SDZ对大肠癌lovo细胞作用24 h,细胞抑制率分别为45%、43%、37%、29%、24%.结论 SDZ体外对大肠癌lovo细胞有生长抑制作用,呈剂量依赖性和时间依赖性.     

败血症患者血液中病原菌分布及耐药性分析

目的了解医院感染败血症患者血液中病原菌分布及耐药性的特征。方法血液培养采用BD BACTEC9050全自动血培养仪，采用复星公司FOUTUNE IMS病原菌鉴定及药敏分析系统。结果分离菌构成比中革兰阴性杆菌处于优势，占64．9％，革兰阳性球菌次之，占18．9％，真菌占16．2％，对革兰阴性杆菌和革兰阳性球菌药敏耐药性相对较低的药物为碳青酶烯类、加酶抑制剂和万古霉素。结论近两年败血症患者血液病原菌分布以革兰阴性杆菌为主，肠球菌属和真菌占一定的比例，病原菌对临床抗菌药物有较高的耐药性。     

AMI患者PCI后心肌灌注水平对心功能及左室重构的影响

目的:探讨急性心肌梗死（AMI）患者经皮冠脉介入术（PCI）后TIMI血流III级时的心肌灌注水平及其对心功能与左室重构的影响。方法: 对36例AMI患者PCI后行经静脉心肌声学造影（MCE）和心脏二维超声检查。①利用心肌声学造影评分（MCS）及室壁运动评分(WMS)分析PCI后心肌灌注情况与室壁运动情况的关系;②根据声学造影积分指数（CSI）将患者分为A、B两组,比较两组的左室射血分数（LVEF）,评估心肌灌注水平对心功能的影响;③根据心脏二维超声结果,比较两组患者术后6个月时左室舒张末直径（LVEDD）及LVEF的变化,进一步评估心肌灌注水平对左室重构的影响。结果: PCI后梗死相关血管TIMI血流均达III级。共152节段与梗死相关血管的再灌注有关。①MCS为0分的18节段中,2个（11.1%）WMS为1~2分;MCS为0.5分的30节段中16个（53.3 %）WMS为1~2分;MCS为1分的104节段中,82个（78.8%）WMS为1~2分;统计学分析显示,PCI后心肌灌注水平与室壁运动呈正相关（P<0.05）。②心肌灌注好的A组LVEF显著大于B组［(52.1±3.4)%,(47.2±2.9)%,P<0.05］。③术后6个月A组的LVEF及LVEDD均无明显变化,B组的LVEF较前有所下降［(47.2±2.9)%,(43.8±4.4)%,P<0.05］,LVEDD较前有所增加［(50.2±2.9) mm,(56.3±3.1) mm,P<0.05］。结论: AMI患者PCI后心肌灌注水平与心功能及左室重构有一定相关性,良好的心肌灌注在一定程度上可以抑制左室重构。     

Model simulation of inflow water to the Baltic Sea based on 129I

The semi-enclosed Baltic Sea represents a vital economic and recreational resource for more than 90 million people inhabiting its coasts. Extensive contamination of this sea by a variety of anthropogenic pollutants has raised the concern of the people in the region. Quantifying seawater inflow is crucial for estimating potential environmental risks as well as to find the best remedial strategy. We present here a model to estimate water inflow from the North Sea to the Baltic Sea by utilizing ¹²⁹I as a tracer. The results predicted inflow range of 230–450 km³/y with best fit value around 330 km³/y from the North Sea to the Baltic Sea during 1980–1999. Despite limited time series data on ¹²⁹I, the model presented here demonstrates a new management tool for the Baltic Sea to calculate inflow water compared to conventional methods (such as salinity, temperature and hydrographic models).     

中药化学成分抗菌活性化学信息学研究

目的:采用化学信息学方法,分析中药化学成分分子结构特征与其抗菌活性的构效关系.方法:采用Cytoscape 和ChemViz计算中药化学成分与目前临床使用的抗菌药物的二维化学结构相似度以及表征细胞渗透性的拓扑参数TPSA(Topolgical Polar Surface Area,拓扑分子极性表而积).继而计算8大类中药化学成分与已知抗菌药物的整体化学结构相似程度.结果:相比其他类别的中药化学成分,黄酮类化合物表现出与现有抗菌药物较好的结构相似度,适宜的细胞渗透性.结合其较好的药物安全性特点,提示这类化合物可作为进一步合成抗菌药物的优选先导化合物骨架结构来源.结论:化学信息学的应用有助于探究中药化学成分分子结构特征与其抗菌活性的构效关系,寻找合适的抗菌药物先导化合物骨架结构.     

Edaravone attenuates ischemia-reperfusion injury by inhibiting oxidative stress in a canine lung transplantation model

Background Previous reports have confirmed that edaravone has protective effects against ischemia-reperfusion （IR） injury of many organs. In this study, we investigated the effect of edaravone on preventing IR injury of the lung in a canine lung transplantation model. Methods Twelve weight-matched pairs of random-bred dogs were randomized into two groups. Within each pair, one dog served as donor and the other as recipient. In the study group, prostaglandin EI（PGE1）（8 μg/kg） was injected into the donor pulmonary artery （PA） before occlusion and the donor lungs were flushed with 1.0 L of LPD solution containing edaravone （10 mg/kg） and stored in the same LPD solution at a temperature of 1℃for 8 hours. The left single lung transplantation was then performed and recipients received intravenous injection with edaravone （10 mg/kg） at the onset of reperfusion. In the control group, edaravone was substituted by the same volume of sterile saline solution. Another six dogs were obtained as normal control group in which left lungs were dissected after thoracotomy without an IR injury. One hour after reperfusion, or after dissection of the left lung, the right lung was excluded from perfusion and ventilation after which, cardiopulmonary parameters were measured. Wet/dry ratios, malondialdehyde （MDA） and myeloperoxidase （MPO） levels were assessed and histological analysis of lung tissue performed at the same time. Results All animals survived until the end of the experiment. The study group showed significantly decreased wet/dry ratios （treated： （74.1±4.2）% vs control： （86.8±5.2）%, P 〈0.01）, MDA levels （treated： 0.50±0.08 vs control： 0.88±0.15, P 〈0.01） and MPO activity （treated： 0.23±0.05 vs control： 0.43±0.07, P 〈0.01） compared to the control group two hours after occlusion of the right side. In the control group, pulmonary vascular resistance （PVR） was increased markedly and arterial oxygen partial pressure deteriorated significantly after     

Neutralization effects of antibody elicited by chimeric HBV S antigen viral-like particles presenting HCV neutralization epitopes

Hepatitis C virus (HCV) infection is a major public health problem despite effectual direct-acting antivirals (DAAs) therapy. Development of a prophylactic vaccine is essential to block spread of HCV infection. The HBV small surface antigen (HBsAg-S) can self-assemble into virus-like particles (VLPs), has higher immunogenicity and is used as a vaccine against HBV infections. Chimeric HBsAg-S proteins with foreign epitopes allow VLP formation and induce the specific humoral and cellular immune responses against the foreign proteins. In this study, we investigated the immune responses induced by chimeric VLPs with HCV neutralizing epitopes and HBV S antigen in mice. The chimeric HCV-HBV VLPs expressing neutralizing epitopes were prepared and purified. BALB/c mice were immunized with purified chimeric VLPs and the serum neutralizing antibodies were analyzed. We found that these chimeric VLPs induced neutralizing antibodies against HCV in mice. Additionally, the murine serum neutralized infections with HCV pseudoparticles and cell-cultured viruses derived from different heterologous 1a, 1b and 2a genotypes. We also found that immunization with chimeric VLPs induced anti-HBsAg antibodies. This study provides a novel strategy for development of a HCV prophylactic neutralizing epitope vaccine and a HCV-HBV bivalent prophylactic vaccine.     

新型冠状病毒肺炎期间骨盆髋臼骨折诊疗专家共识

自2019年12月以来,新型冠状病毒(2019-nCoV)感染的肺炎(COVID-19)疫情暴发并向全国蔓延。相较于严重急性呼吸综合征(SARS)冠状病毒,2019-nCoV致死率更低,但却具有更高的传染性和致病力。截至2020年2月22日24时,累计报告确诊病例76936例(武汉36174例)[1];截至2月11日,全国医务人员感染3019例[2],远远超过了"非典"疫情感染人数。骨盆、髋臼骨折是创伤救治中最为复杂的病情之一,其发生率约占全身骨折的3%[3]。骨盆、髋臼骨折常发生于高能量损伤,容易合并大出血、重要脏器损伤等严重情况,病死率一直居高不下[4]。病毒流行为疫区骨盆、髋臼骨折院前急救、急诊诊疗、手术策略、麻醉和围手术期管理等医务工作带来了巨大挑战。在疫情流行期间,为了降低骨盆、髋臼骨折救治过程中患者之间、患者与医务人员之间、医务人员相互之间交叉感染的风险,提高骨折预后水平,需要通过多学科协作,统筹传染病和骨盆、髋臼骨折的诊疗方案,制定严密的诊疗计划,降低疾病的致残率和致死率。为响应党中央号召,科学战"疫",坚决打赢这场疫情防控的人民战争、总体战、阻击战,根据《新型冠状病毒感染的肺炎诊疗方案(试行第六版)》,结合肺炎防治和创伤救治相关诊疗指南[5,6,7,8,9,10],特制定此诊疗专家共识,以规范COVID-19疫情期间骨盆、髋臼骨折的诊疗,提高治愈率,降低交叉感染的风险。     

64层螺旋CTPA对肺动脉栓塞的诊断价值(附25例分析)

目的评价64层螺旋CTPA及三维成像技术对肺动脉栓塞的诊断价值。方法收集25例经64层SCTPA检查确诊的肺动脉栓塞;回顾性分析其成像条件及薄层容积图像,MPR、MIP、SSD、VR等后处理方法对肺动脉及其栓子的显示情况,并观察肺动脉栓塞征象。结果25例SCTPA图像质量均满足诊断需要;薄层容积图像可清晰显示肺动脉及其各级分支,显示率为100%;MPR、MIP、SSD、CR等后处理法有助于多角度、多方位显示栓子。PE主要累及左右肺动脉等大分支,占90%,其中中央型栓塞18,周围型栓塞7例。栓塞直接征象为栓子形成充盈缺损;间接征象包括肺梗死、胸腔积液、心包积液、膈肌抬高和右心增大等。结论64层螺旋CT极大提高了SCTPA这一技术的临床应用价值,是目前诊断肺动脉栓塞最为快捷、安全、可靠的影像方法。