半套式PCR和DNA探针技术检测Q热立克次体

目的 建立半套式PCR和DNA探针技术由于检测Q热立克次体。方法 依据已知的Q热立克次体的16S和23S rRNA基因及其间区的序列。设计3条引物,建立了扩增16S－23S mRNA基因间区的半套式PCR方法,并将扩增片段制成探针,建立了斑点杂交技术。结果 10株Q热立克次体分离株均可扩增出572bp的PCR条带,而对照菌都为阴性,此半套式PCR方法的灵敏度高达1pg;用九里株扩增片段标记后制成的探针,与10个Q热立克次体分离株的全DNA呈阳性杂交,对照菌为阴性,此杂交方法的灵敏度为0．5ng。结论 基于Q热立克次体16S－23S rRNA基因间区的半套式PCR和DNA探针技术具有较好的特异性和灵敏度,可联合应用于Q热的病原学诊断。     

内毒素血症小鼠巨噬细胞膜脂改变对CD14表达及TNFα分泌的影响

目的 以小鼠内毒素血症为模型,研究小鼠腹腔巨噬细胞膜脂改变,及其对受体CD14表达及血浆TNFα产生的影响。方法 48只小鼠分为3组。①对照线（n=8）;②内毒素血症组：小鼠尾静脉注射LPS,200μg/只,分1、3、5、8 4个观察时相点,每个时相点8只动物;③脂质体预处理组（n=8）：小鼠腹腔注射卵磷脂脂质体1ml(1mg/ml),18h后尾静脉注射LPS,200μg/只,5h后观察巨噬细胞膜脂成分、膜流动性及CD14阳性率、血浆中TNFα含量的变化。结果 LPS刺激下,巨噬细胞主要膜脂成分降解,细胞膜荧光偏振度、微粘度和分子排列有序系数增加,膜流动性显著降低;CD14阳性率增加,5h达高峰,8h降至正常水平;TNFα含量显著升高,3h达高峰,8h降至正常水平。磷脂脂质体预处理后,膜损伤减轻,CD14阳性率及TNFα含量均明显降低。结论 内毒素血症小鼠腹腔巨噬细胞膜磷脂降解,细胞膜从相对比较流动变为相对固化,这可能是膜受体CD14阳性率增加,TNFα产生增多的重要原因。脂质体预处理能修复膜损伤,减少膜CD14表达,减轻炎症反应。     

伤寒史学研究成果简述

建国以来有关伤寒史学的研究成果表现在相关人物、相关著作、相关事件、学术关系及阶段特征五个方面。前期研究成果 ,尚不足以充分展示伤寒学历史地位和现实意义 ,更无充分依据昭示其未来之发展方向 ,因而有必要继续深入开展伤寒史学研究     

柱切换HPLC法测定腹水中左旋氧氟沙星的含量

目的：应用柱切换ＨＰＬＣ技术直接测定腹水中左旋氧氟沙得含量。方法：采用磷酸二氢钾缓冲溶液（ＰＨ２．２）－０．０５ｍｏｌ／Ｌ四丁溴化化铵（１００：４）为预处理流动相,经μＢｏｎｄａｐａｋ Ｃ１８柱预处理后,切换到Ｉｒｒｅｇｕｌａｒ＿ＨＣ１８分析柱,以甲醇－磷权二氢钾缓冲溶液（ＰＨ２．２）－０．０５ｍｏｌ／Ｌ四丁基溴化铵（３０：７０：４）为分析流动相进行分离测定,紫外线皮长为２９４ｎｍ。结果：左旋氧氟     

The mutation of insulin receptor substrate-1 gene in Chines e patients with non-insulin-dependent diabetes mellitus

Objective To identify the relationship between mutation in the insulin receptor substrate-1 (IRS-1) gene and the incidence of non-insulin-dependent diabetes mellitus (NIDDM) in the Chinese population. Methods Samples were obtained from 68 Chinese patients with NIDDM and 68 control subjects. The +1700～+4437 bp fragment of the IRS-1 gene was screened by polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) analysis. All SSCP variations were submitted to DNA sequence analysis. Results Two amino acid variations ［GGG →AGG(G(971) R) and CCT→TCT(P(1079) S)］ and 3 silent mutations ［GAT→GAC(D(422) D), CCA→CCC(P(737) P) and GCA→GCG(A(804) A)］ were identified, among which the CCA→CCC(P(737) P) and CCT→TCT(P(1079) S) have not been previously reported. All five variations were found in Chinese patients with NIDDM, while GCA→GCG(A(804) A) was the only one found in control subjects. The overall incidence of the five variations in Chinese patients with NIDDM were much higher than that in control subjects (38.2% vs 7.4%, χ(2)=18.42,P<0.01). The most common polymorphism in the Chinese population was GCA→GCG(A(804) A), and its frequency was significantly higher in Chinese patients with NIDDM than in controls (26.5% vs 7.4%, χ(2)=8.84,P<0.01). The homozygotes of the variation in patients with NIDDM and control subjects were 8.8% and 1.5%, respectively (χ(2)=2.41,P>0.05). Conclusion These results indicate that there may be a relation between these nucleotide variations of IRS-1 gene and Chinese patients with NIDDM     

肾功能分级在成人心脏术后肾功能衰竭的应用

Objective To evaluate the ability of the RIFLE classification to predict hospital mortality in adult patients who underwent cardiac surgery. Methods From October Ist 2006 to December 31st 2006, five hundred and nine adult patients who underwent coronary artery bypass grafting and/or valve operation were enrolled in this study. Renal function was assessed daily according to the RIFLE classification, meanwhile, APACHE Ⅱ score and SOFA score were also evaluated, as well as the maximum scores were recorded. Results Mean duration of ventilation support was 18(14 - 19) hours, the time of ICU stay was 1.4 ± 1.0 days, and the time of postoperative hospital stay was 12. 0(10.0- 15.0) days. 167 patients (32. 8%) incurred postoperative ARF according to the RIFLE classification. The overall mortality was 4. 3% (22/502). A significant increase (P < 0. 01) was observed for mortality based on RIFLE classification. By applying the area under the receiver operating characteristic curve, the RIFLE classification had more powerful discrimination power [0. 933, (95% CI 0. 872 -0. 995) ,P <0. 001]. Conclusions ARF is one of the major complications in postcardiotomy patients. Analytical data suggested the good discriminative power of the RIFLE classification for predicting inpatient mortality of adult postoperative patient with ARF, and the RIFLE classification is simple and practically performed. According to the RIFLE classification, patients with RIFLE class I or class F incur a significantly increased risk of in-hospital mortality compared with those who never develop ARF.     

成人散发性病毒性肝炎病原学分型与HGV重叠感染

目的 了解成人急性散发性病毒性肝炎的病原学分型及庚型肝炎病毒感染状况。方法 利用ＥＬＩＳＡ和逆转录聚合酶链反应对病人血清进行检测。结果 成人急性肝炎中以乙型肝炎所占比例最大,占３７．１６％;其次甲型肝炎占１８．９２％,另有１８．２４％为非甲～庚型肝炎。结论 庚型肝炎病毒存在明显的重叠感染,与乙型和丙型肝炎病毒重叠感染率较高。     

环磷酰胺对大鼠脑缺血再灌流损伤的免疫保护

目的 观察免疫抑制剂对脑缺血再灌流损伤的影响及可能作用机制。方法 采用４－ＶＯ法制作大鼠脑缺血再灌流损伤模型,观察环磷酰胺预治疗大鼠外周血白细胞数,海马ＣＡ１区的计量病理,海马区内二醛含量和Ｎａ＾＋,Ｎ＾＋－ＡＴＰ酶活性的变化。结果 与对照组比较,预治疗组大鼠外周血内白细胞数明显降低,ＣＡ１区正常神经元计数增加,结论 环磷酰胺可ＡＴ酶活性升高而ＭＤ芩明显减低。结论 环磷酰胺可减轻大鼠离缺血再灌流损     

含CpG寡聚脱氧核苷酸对肿瘤抗原肽P815AB的免疫佐剂效应研究

目的 以肿瘤抗原Ｐ８１５ＡＢ抗原肽为模式抗原,研究ＣｐＧ寡聚脱氧核苷酸（ＣｐＧ ＯＤＮ）对该肿瘤抗原表位肽的免疫佐剂效应。方法 用［＾３Ｈ］－ＴｄＲ掺入法研究ＣｐＧ ＯＤＮ体外刺激小鼠脾细胞增殖效应,用ＥＬＩＳＰＯＴ和［＾３Ｈ］－ＴｄＲ释放法检测不同构成形式的疫苗在体诱发特异性ＣＴＬ应答的效应。结果 ＯＤＮ１８２６在体外可刺激小鼠脾细胞显著增殖,经４次免疫,在“肽＋ＯＤＮ１８２６＋ＩＦＡ”组中,４     

小鼠肥大细胞瘤P815特异抗原P1A的克隆与表达

目的 克隆小鼠肥大细胞瘤Ｐ８１５细胞株的ＰＩＡ基因以制备肿瘤ＤＮＡ疫苗,方法 用ＲＴ－ＰＣＲ方法制备ＰＩＡ基因,以哺乳细胞高效表达质粒ｐＣＩ－ｎｅｏ为载体,构建重组ＤＮＡ疫苗。重组体用克隆位点上游和下游的Ｔ７和Ｔ１３启动了序列为测序引物,用自动测序仪测定鉴定克隆的正确性。再将鉴定过的重组质粒用磷酸钙法转化２９３细胞,用ＲＴ－ＰＣＲ法鉴定转化细胞中ＰＩＡ基因的表达。结果 正确构建了ＰＩＡ／ｐＣＩ－ｎ