恐惧应激对大鼠睾丸中Annexin5表达的影响

目的观察大鼠应激条件下睾丸组织中Annexin5表达的变化。方法建立SD大鼠的恐惧应激模型，分别取急性应激、急性对照、慢性应激、慢性对照SD大鼠睾丸，免疫组织化学和Western blotting分析Annexin5免疫定位和蛋白表达。结果急性应激组与对照组相比，免疫组化显示Annexin5都定位在间质细胞、支持细胞和成熟的精子头部，Western blotting分析发现Annexin5的表达降低了10．8％；慢性应激组与对照组相比，免疫组化结果显示Annexin5在精子头部的定位逐渐减少，应激1w和2w组，未见到Annexin5定位于精子头部，到了应激的3w和4w，发现Annexin5重新定位于精子的头部，而在间质细胞和支持细胞上的定位模式没有显著变化。Western blotting分析发现Annexin5的表达降低了17．4％。结论大鼠睾丸Annexin5参与了应激过程，急性应激条件下Annexin5表达降低。随着应激时间的延长，在慢性应激条件下，Annexin5的表达由减少之后而开始有缓慢增加。     

The <Emphasis Type="Italic">Arg</Emphasis>194<Emphasis Type="Italic">Trp</Emphasis> polymorphism in DNA repair gene XRCC1 and the risk for sporadic late-onset Alzheimer's disease

Abstract Alzheimer's disease (AD) is defined pathologically by the presence of β-amyloid plaques, neurofibrillary tangles and extensive neuronal loss. Evidence indicates that increased DNA damage may contribute to neuronal loss in AD. Recently, it has been shown that in AD neurons have a reduced capacity for some types of DNA repair. Polymorphisms in DNA repair genes may be associated with differences in repair efficiency of DNA damage. Variants of several DNA repair genes, including the base excision repair gene XRCC1, have been described previously. We hypothesised that Arg194Trp polymorphism of XRCC1 gene may contribute to genetic susceptibility for AD. In order to test this hypothesis, we investigated Arg194Trp polymorphism at the XRCC1 gene in the DNA samples of 98 patients with AD and 95 healthy subjects. The frequency of the Trp allele was more pronounced among cases (11.2%) compared with controls (5.8%). On combining the homozygous and heterozygous variants of each codon, the variants seemed to be at twofold risk of AD, although the risk estimates were not statistically significant (OR=1.95, 95% CI 0.88–4.34, p=0.09). In addition, the 194Trp allele revealed a borderline significance (OR=2.05, 95% CI 0.96–4.37, p=0.056). According to our results, it may be speculated that the polymorphic variants of XRCC1 codon 194 have a role in the development of AD.     

SARS相关冠状病毒S1融合蛋白的原核表达与纯化

目的：克隆严重急性呼吸综合征相关冠状病毒(SARS-CoV)S1蛋白编码DNA，构建原核表达质粒pGEX-5T／S1，并诱导表达谷胱甘肽巯基转移酶融合蛋白。方法：采用PCR方法扩增合成S1片段，并克隆入载体中，经双酶切鉴定和测序后把S1序列定向插入原核表达载体pGEX-5T的多克隆位点，转化大肠杆菌K802，将纯化后的融合蛋白用于SARS—CoV抗体阳性血清的检测。结果：GST—S1融合蛋白以可溶形式表达，纯化后的蛋白用ELISA法检测，结果与对照相符。结论：成功诱导原核表达并纬化出融合蛋白S1，为将其应用于SARS—CoV的特异性检测和亚单位疫苗研究奠定了基础。     

糖尿病大鼠肾组织中单核细胞趋化蛋白-1的表达及意义

目的：探讨单核细胞趋化蛋白-1(MCP—1)在糖尿病大鼠肾组织中的表达及其意义。方法：用链脲佐菌素制备糖尿病大鼠模型。采用免疫组化及多媒体彩色病理图文分析系统观察肾小球及肾小管中MCP-1的表达。结果：糖尿病大鼠肾小球及肾小管中MCP—1的表达显著上调。结论：MCP-1在糖尿病肾病发病机制中发挥重要作用。     

褪黑素对胰岛素抵抗大鼠肾皮质内皮素、内皮型一氧化氮合酶平衡的影响

目的 研究褪黑素对胰岛素抵抗(IR)大鼠肾皮质内皮素1(ET-1)、内皮型一氧化氮合酶(eNOS)表达的影响.方法 高糖饲料喂养SD大鼠6周复制IR大鼠模型.成模后用药组(MEL组)予褪黑素10mg·kg-1·d-1灌胃,用药6周.未用药IR大鼠模型为对照(IR组).免疫组化、RT-PCR方法 检测肾皮质ET-1、eNOS蛋白和mRNA表达的改变.结果 与IR组相比,褪黑素组大鼠动脉血压(ABP)、血清TG、游离脂肪酸(FFA)、丙二醛(MDA)、胰岛素(Ins)和HOMA-IR降低(P＜0.01),血清HDL-c、超氧化物歧化酶(SOD)升高(P＜0.01).HE染色MEL组大鼠肾皮质病变明显减轻.MEL组ET-1蛋白及mRNA明显低于IR组、eNOS蛋白及mRNA明显高于IR组,差别均有显著意义(P＜0.05).结论 在IR早期应用褪黑素(10mg·kg-1·d-1)治疗可以减轻IR大鼠肾皮质ET-1与eNOS平衡的异常,对肾皮质血管内皮功能具有一定保护作用.     

血清1,5-脱水葡糖醇的测定及在糖尿病诊断中的应用

目的:应用血清1,5-脱水葡糖醇(1,5AG)的酶动力学法,探讨其在糖尿病诊断中的应用价值。方法:应用全自动生化分析仪,采用全酶法测定健康人和糖尿病患者血清1,5AG的含量。结果:该法测定血清1,5AG精密度好,高、低值混合血清的CV分别为1.47％和1.70％;回收率分别为98.28％、101.32％和97.64％、103.36％,平均回收率为100.15％;在1,5-AG浓度为360μmol/L内呈良好的线性;60例糖尿患者血清1,5AG(5.38-59.54μmol/L)明显低于健康人(60.89～269.79μmol/L),P<0.01。结论:该方法重复性好,准确度高,工作试剂在2 d内稳定,适合常规实验室开展;血清1,5AG测定有助于糖尿病的诊断和治疗过程的监控。     

胃癌螺旋CT与病理、nm23-H1蛋白表达的相关性研究

目的 探讨胃癌螺旋CT征象与手术病理及nm23-H1蛋白表达间的关系.资料与方法 对65例胃癌行低张力水充盈螺旋CT三期增强扫描,所有病例均行手术切除,术后标本采用免疫组织化学SP法检测肿瘤组织中nm23-H1蛋白表达.将螺旋CT诊断结果 与病理结果 、nm23-H1蛋白表达进行对照.结果 65例胃癌TNM分期CT的准确性为80.0%(52/65),nm23-H1蛋白阳性表达率为50.8%(33/65).CT像上的病灶大小、浆膜侵犯、淋巴结转移、TNM分期与病理结果 一致性良好,与nm23-H1蛋白阳性表达率均密切相关(P＜0.05).结论 螺旋CT可较准确地反映胃癌增殖、浸润转移的病理学及生物学特性.     

高脂血症对血浆t-PA和PAI-1活性的影响

目的 探讨高脂血症患者的纤溶系统的变化。方法 选取正常人及高脂血症者，测定其血脂、血浆t—PA和PAI-1活性。结果 高脂血症组比正常人血浆t—PA活性下降，而PAI-1的活性有升高；在TG及LDL-C升高者，其血浆t—PA活性比正常人血浆t—PA活性降低，而血浆PAI-1活性比正常人升高；单纯TC升高者血浆PAI-1活性接近正常人组，而血浆t—PA活性比正常人组明显降低。结论 高脂血症对纤溶系统有一定的影响，其中甘油三酯和LDL-C对纤溶系统的影响更明显，而胆固醇对纤溶系统也有一定的影响。     

Influence of bradykinin B1 and B2 receptors in the immune response triggered by renal ischemia–reperfusion injury

Bradykinin B1 receptors are exclusively expressed in inflamed tissues. For this reason, they have been related with the outcomes of several pathologies. Ischemia–reperfusion injury is caused by the activation of inflammatory and cytoprotective genes, such as macrophage chemoattractant protein-1 and heme oxygenase-1, respectively. This study was aimed to analyze the involvement of bradykinin B1 and B2 receptors (B1R and B2R) in tissue response after renal ischemia–reperfusion injury. For that, B1R (B1−/−), B2R (B2−/−) knockout animals and its control (wild-type mice, B1B2+/+) were subjected to renal bilateral ischemia, followed by 24, 48 and 120 h of reperfusion. At these time points, blood serum samples were collected for creatinine and urea dosages. Kidneys were harvested for histology and molecular analyses by real-time PCR. At 24 and 48 h of reperfusion, B1−/− group resulted in the lowest serum creatinine and urea levels, indicating less renal damage, which was proved by renal histology. Renal protection associated with B1−/− mice was also related with higher expression of HO-1 and lower expression of MCP-1. In conclusion, the absence of B1R had a protective role against inflammatory responses developed after renal ischemia–reperfusion injury.