SNAP-25基因克隆及其在大肠杆菌中的表达与鉴定

目的：构建SNAP-25基因的原核表达载体pET22b—SNAP-25,对表达产物活性进行初步鉴定。方法：根据GenBank中已报道的SNAP-25基因序列,设计特异引物,通过RT—PCR从小鼠全脑组织总RNA中得到基因。将全长基因克隆至原核表达载体pET22b中,重组质粒转化大肠杆菌E.coli如BL21感受态细胞,IPTG诱导表达,表达产物经Ni—NTA亲和层析进行纯化。通过SDS-PAGE和免疫印迹对其进行鉴定,并对该蛋白进行活性的初步分析。结果与结论：成功构建了原核表达载体pET22b—SNAP-25,Western印迹证实重组蛋白获得表达。表达的重组蛋白与A型肉毒毒素混合反应,经SDS—PAGE验证,重组蛋白具有被毒素特异性切割的活性。     

The backpack run test: a model for a fair and occupationally relevant military fitness test

Our purpose in this investigation was to develop and validate a theoretical model for a backpack run test based on how fast one can run 2 miles while wearing a backpack. Using actual unloaded (no backpack) 2-mile-run test data from 59 male service academy cadets, we calculated the average oxygen cost during the run, the equivalent cost if wearing additional weight, and the corresponding estimated run time with the backpack. The correlations between body weight and loaded (backpack weight = 30 kg) run times (r = 0.55 [p < 0.05] and r = 0.12 [p > 0.05], respectively) demonstrate that the bias against heavier runners is eliminated with the backpack run. Given that the backpack run test requires only standard-issue equipment, demonstrates clear occupational and health-related fitness relevance, predicts no apparent body-size bias, and measures work- and health-related components of fitness, we recommend that the military services consider the present data when developing or modifying tests of physical fitness.     

肠出血性大肠杆菌O157∶H7毒力蛋白NleB1的表达、纯化及多克隆抗体的制备与鉴定

目的：构建肠出血性大肠杆菌（ EHEC） O157∶H7毒力蛋白nleB1基因的原核表达载体,诱导表达重组蛋白,制备NleB1的抗血清。方法从EHEC O157∶H7 EDL933株的全基因组中PCR扩增出编码nleB1的990个碱基序列,构建原核表达载体pET24a-nleB1,将构建的重组表达载体的质粒pET24a-nleB1转化至大肠杆菌BL21（DE3）感受态细胞中,利用异丙硫半乳糖苷（IPTG）诱导融合蛋白His-NleB1表达,并利用镍柱进行亲和层析纯化和切胶纯化,以纯化后的融合蛋白His-NleB1为抗原免疫BALB/c小鼠,获得抗血清。 Western印迹和ELISA法鉴定获得的抗血清。结果成功构建了pET24a-nleB1原核表达载体,纯化得到融合蛋白His-NleB1,进而免疫BALB/c小鼠制备多克隆抗体,Western印迹和ELISA法证实多克隆抗体制备成功。结论成功获得了EHEC O157∶H7的毒力蛋白NleB1的多克隆抗体,为研究EHEC O157∶H7毒力蛋白NleB1的功能奠定了基础。     

The rights of migrants to the identification of their dead: an attempt at an identification strategy from Italy

International Journal of Legal Medicine - Europe is turning a blind eye on a humanitarian disaster unfolding at its doorsteps, with thousands of migrants dying unidentified in Mediterranean waters....     

大肠杆菌基因组水平蛋白质-RNA相互作用初步研究

目的初步研究大肠杆菌中基因组水平的蛋白质-RNA相互作用（protein-RNA interactions,PRI）。方法通过RNA酶消化细菌裂解液,提取与蛋白质相互作用的RNA片段,构建cDNA文库,进行高通量测序,并通过生物信息学分析获得与蛋白质结合的转录本。结果获得了与蛋白质结合的3193条转录本,涉及2234个mRNA、47个sRNA（small regulatory RNAs）、39个tRNA、11个rRNA以及862个基因间区（intergenic region,IGR）。结论初步获得大肠杆菌中与蛋白质相互作用的转录本信息,为进一步开展PRI研究提供了支持。     

Preparation of a DNA probe for thermolabile toxin and its use in the identification of enterotoxic Escherichia coli

Enterotoxic Escherichia coli (ETEC) having a gene for thermolabile toxin were identified by the hybridization technique. HindIII-SmaI fragment with 423 basic pairs (dp) including subunit portions A and B of the gene for thermolabile toxin (elt) was used as the DNA probe.     

Patient satisfaction in military medicine: status and an empirical test of a model

The Department of Defense (DoD) is concerned about how well military medical treatment facilities in the military health system perform. Patient expectations, attitudes, and health care use have been examined in numerous studies; the results are fairly consistent. Eligible beneficiaries report moderate satisfaction with the health care received. In 1994-2001, annual DoD and monthly ambulatory patient surveys were conducted in military medical treatment facilities. The DoD surveys document how patients perceive the care provided. The obvious research concerns are: requirements for conducting surveys; who should be surveyed: eligible beneficiaries or actual users; when; where; representative sample; how often to conduct assessment; data collection methods; analytic schemes; overall trends; predictors of satisfaction; use of results; and timeliness of findings. This study examines these issues and analyzes raw data from selected annual DoD and monthly ambulatory surveys. The overall level of perceived satisfaction has been "good" over the years surveys were used. The model demonstrated the use of examining demographic and attitudinal components of patient satisfaction in military medical facilities.