重组沙门菌表达结核分枝杆菌ESAT-6及其特异性免疫应答分析

Objective To determine the immune responses induced by recombinant Salmonella ty-phimurium expressing the secreting antigen ESAT-6 of Mycobacterium tuberculosis. Methods ESAT-6 cod-ing gene was cloned and identified by PCR and sequencing. Prokaryotic expression plasmid pYA33-esat car-rying the ESAT-6 coding sequence was constructed firstly and electro-transformed into an attenuated strain X4550 of Salmonella typhimurium, the recombinant bacteria was named as X4550(33-esat). C57BL/6 mice were immunized intranasally (I. N) with 108 CFU recombinant bacteria at day 0 and 18. Cells from spleen, lung, mesenteric lymph node (MLN) and Peyer's patch (PP) were collected from mice after second immu-nization, and the specific IFN-γ-secreting cells and IL-4-secreting cells were detected by ELISPOT assay u-sing ESAT-6 peptide as stimulus. Furthermore, CTL effects were in vivo evaluated by CFSE assay. Results The results showed that cellular immune responses specific for ESAT-6 could be detected by ELISPOT assay. In lung and PP cells, immune responses against ESAT-6 were biased toward Th1 type, the frequency of IFN-γ-secreting cells was much higher than that of IL-4-secreting cells. In splenocytes and MLN cells, the anti-gen specific immune responses acted as Thl and Th2 balance, the frequency of IFN-γ-secreting cells was close to that of IL-4-secreting cells. CFSE assay indicated that recombinant bacteria could induce the high level of CTL effects specific for ESAT-6 peptide. Conclusion These results suggested that recombinant Sal-monella typhimurium X4550(33-esat) not only can induce cellular immune responses, but also can elicit specific CTL responses after I. N immunization. It also provided the useful information for the control of infec-tious disease of tuberculosis.     

环甲膜切开术在颅脑损伤救治中的应用

环甲膜切开术在颅脑损伤救治中的应用兰州医学院兰化教学医院神经外科（７ｓ００６０）赵卫忠，张文科，王明泉，李天郁，许辉呼吸道的通畅是神经外科治疗中的一个重要问题，尤其在急症的抢救中，往往由于周围或中枢性呼吸困难，多需紧急气管切开。自１９８６年，我们采用...     

宫颈刮片快速染色法

妇女病普查中宫颈刮片染色检查法是普遍采用的,通过 宫颈刮片的染色检查,可以反映卵巢功能,亦可诊断炎症及 肿瘤,且可在病变早期发现。根据我院多年来妇女病普查的 经验总结出来操作简便快速的方法——宫颈细胞快速染色 法。     

解热镇痛药到底该怎么选

冬季是大人和孩子感冒的多发季节。许多人在感冒初期为了免去医院的麻烦,便喜欢到就近的药店买一些解热镇痛药,但是含对乙酰氨基酚的解热镇痛药剂型非常多,有片剂、胶囊剂、咀嚼片、混悬剂、糖浆剂、滴剂、栓剂、分散片、泡腾片等,到底该如何选呢？     

灌注法构建小肠黏膜全层去细胞外基质

背景：对于解剖层次复杂的细胞外基质,单纯浸泡法则很难达到理想的去细胞效果.目的：观察灌注法去细胞技术制备小肠黏膜全层去细胞外基质的效果. 方法：采用灌注法去细胞技术,经成年雄性新西兰大白兔小肠黏膜上动脉灌注去离子剂后处理获得小肠黏膜全层去细胞外基质.采用MTT法检测小肠黏膜全层去细胞外基质浸提液（实验组）或含体积分数20%小牛血清的DMEM（对照组）与1月龄新西兰大白兔骨髓间充质干细胞共培养后的细胞相对增殖率. 结果与结论：①大体观察：小肠黏膜上动脉经灌注30 min后变得苍白透明,经2 h灌注后肠段变得苍白剔透,清楚可见脉管纹理.②组织学与扫描电镜观察：灌注法构建的小肠黏膜全层去细胞外基质的去细胞程度均匀,胶原纤维未受损,孔隙多,孔隙率为（86.72±2.98）%.③M TT 检测实验：培养2,4,7 d,实验组细胞A值明显高于对照组细胞A值（P〈0.05）,且实验组细胞相对增殖率均〉1.表明采用灌注法构建小肠黏膜全层去细胞外基质可操作性强,安全性好,且小肠黏膜全层去细胞外基质对骨髓间充质干细胞无毒性,并有一定的促生长作用.     

DIP内镜评分方案在评价行鼻内镜手术的慢性鼻-鼻窦炎患者病情方面的应用

目的探讨DIP内镜评分方案在慢性鼻-鼻窦炎(CRS)行鼻内镜手术中的价值。方法选取2013年9月-2015年2月该院收治行鼻内镜手术的82例CRS患者为研究对象,术前进行Lund-Mackay CT评分、鼻腔鼻窦结局测量20条(SNOT-20)及视觉模拟量表(VAS)评分,术中录制鼻内镜检查录像。术后3个月再次进行SNOT-20和VAS评分,再次录制鼻内镜录像。由两名医生作为评分者对所有录像进行DIP及Lund-Kennedy内镜评分,采用组内相关系数(ICC)来比较两者的信度,分析各评分结果之间的相关性。结果术后SNOT-20、VAS评分、DIP及Lund-Kennedy内镜评分均明显低于术前(P0.05)。术前DIP内镜评分与Lund-Kennedy内镜评分、Lund-Mackay CT评分均成正相关(P0.05),与SNOT-20及VAS评分均无明显的相关性(P0.05)。术后DIP内镜评分与Lund-Kennedy内镜评分成正相关(P0.05),与SNOT-20及VAS评分均无明显的相关性(P0.05)。手术前后DIP内镜评分的评分者信度及重测信度的ICC均高于Lund-Kennedy内镜评分。结论 DIP内镜评分在CRS患者鼻内镜手术前后均有较高的信度,与其他客观评估结果呈正相关,而与主观评分结果无明显相关,故需从主观和客观两个方面来综合评估CRS的病情及疗效。     

应用D7S820基因座研究中国8个地区汉族人群的亲缘关系

收集D7S820基因座的8个不同地区汉族的相关遗传资料。应用亲缘系数(I)和遗传距离(D)研究八个汉族群体间的遗传关系，并根据遗传距离绘制遗传树。结果显示，成都、贵州和浙江汉族为一群，西安、太原、河北和长白山汉族为一群，而北京汉族为独立的一群。