肠缺血再灌注大鼠肺及血液白细胞介素6、白细胞介素8和肿瘤坏死因子α变化与辅酶Q10的干预

目的:观察大鼠肠缺血再灌注时肺及血液白细胞介素6、白细胞介素8和肿瘤坏死因子α的变化以及辅酶Q10的影响。方法:实验于2006-12/2007-02在滨州医学院药理学实验室和免疫学实验室(山东省重点学科三级实验室)完成。①实验分组:清洁级健康Wistar大鼠30只,体质量290～390g,随机数字表法分为假手术组、肠缺血再灌注组,辅酶Q10处理组(缺血再灌注 辅酶Q10),每组10只。②实验方法:建立肠缺血再灌注模型,钝性分离肠系膜上动脉的根部,假手术组不作其他处理;肠缺血再灌注组再灌前30min时经股静脉注入生理盐水10mL/kg;辅酶Q10处理组再灌前30min时经股静脉注入辅酶Q1010mg/kg。③实验评估:酶联免疫吸附法(ELISA)检测各组动物血液、肺组织匀浆及肺泡灌洗液中的白细胞介素6、白细胞介素8和肿瘤坏死因子α含量;光镜下观察肺组织形态学变化。肺泡灌洗液沉渣进行白细胞计数和分类。结果:纳入大鼠30只,均进入结果分析。①肺组织形态学变化:假手术组肺组织无病理改变;肠缺血再灌注组肺间质明显水肿,中性粒细胞浸润,有少量的出血和纤维蛋白渗出;辅酶Q10处理组肺间质轻度水肿,少量的中性粒细胞。②肺泡灌洗液白细胞计数组间无显著性差异,多形核细胞分类肠缺血再灌注组明显高于假手术组(P<0.05),而辅酶Q10处理组与其他两组间差异无显著性意义(P>0.05)。③各组动物血液、肺组织匀浆及肺泡灌洗液中的白细胞介素6、白细胞介素8和肿瘤坏死因子α含量:与假手术组比较,肠缺血再灌注组血液、肺组织匀浆及肺泡灌洗液中白细胞介素6、白细胞介素8和肿瘤坏死因子α显著升高(P<0.05或P<0.01)。与肠缺血再灌注组比较,辅酶Q10处理组血液、肺组织匀浆和肺泡灌洗液中白细胞介素6含量显著降低(P<0.05);白细胞介素8和肿瘤坏死因子α含量在血液中显著降低(P<0.05);而在肺组织匀浆中含量未见显著降低(P>0.05);在肺泡灌洗液中含量亦未见显著降低(P>0.05)。结论:辅酶Q10可能通过抑制白细胞介素6、白细胞介素8和肿瘤坏死因子α炎性因子的释放,对大鼠肠缺血再灌后肺损伤有一定的保护作用。     

Artificial urinary sphincters: plain radiography of malfunction and complications

Inflatable artificial urinary sphincters provide excellent voluntary continence. Eighty-four consecutive patients underwent implantation of artificial urinary sphincters for intractable urinary incontinence; 33 patients had 58 episodes of sphincter malfunction, and eight patients had eight complications involving a functional prosthetic sphincter. Retrospective analysis was performed to determine the value of plain radiography of the pelvis in patients with sphincter malfunction or complication. The cause of malfunction in the majority of patients was a system leak and subsequent loss of hydraulic fluid (31 occurrences; 53%). Plain radiography permitted correct identification of all instances of fluid leakage in patients with opacified prostheses. Plain radiographs were of no value in examining patients with nonopacified prostheses or the complications of cuff erosion or wound infection. Due to the low cost and noninvasive nature of plain radiography of the pelvis, we conclude that it should be used as the initial diagnostic modality in patients with previously opacified but currently dysfunctional artificial urinary sphincters.     

Cytometric detection of DNA amplified with fluorescent primers: applications to analysis of clonal bcl-2 and IgH gene rearrangements in malignant lymphomas

Follicular lymphomas comprise almost two thirds of the US adult non- Hodgkin's lymphomas (NHL) and are the most common malignancy of B- lineage lymphocytes. Polymerase chain reaction (PCR) protocols have been developed to detect the t(14;18) translocation, which juxtaposes the bcl-2 proto-oncogene to the Ig heavy-chain (IgH) gene in 85% of follicular lymphomas and monoclonal rearrangements of the IgH gene in B- cell NHL that lack bcl-2 rearrangements. We used PCR to amplify bcl-2 and IgH rearrangements in DNA from patients with lymphoproliferative disorders and analyzed the products in parallel by gel electrophoresis and flow cytometry, which detected PCR products incorporating fluoresceinated oligonucleotide primers by sequence-specific capture to oligonucleotide-coated magnetic beads. Overall, flow cytometry was superior to electrophoresis of ethidium-bromide-stained agarose gels for detection of products of nested PCR to detect intergenic rearrangements involving bcl-2 and single primer-pair amplification of clonal rearrangement of IgH. Flow cytometric analysis detected bcl-2 translocations in 12 of 13 CD10+ B-cell lymphomas and clonal IgH rearrangements in 14 of 17 monoclonal B-cell populations. In contrast, analysis by gel electrophoresis detected bcl-2 translocations in only 10 of 13 CD10+ and clonal IgH gene rearrangements in only 9 of 17 monoclonal B-cell populations. Flow cytometric analysis was more sensitive than gel electrophoresis and could detect a 16-fold greater dilution of a bcl-2-amplified product than gel electrophoresis. Similarly, flow cytometry could detect an amplification product when template DNA was diluted 10,000-fold, whereas gel electrophoresis only detected amplification products when template was subjected to dilution between 100- and 1,000-fold. This shows the utility of flow cytometry for the analysis of DNA amplification products incorporating fluorochrome-labeled primers as a rapid, objective alternative to conventional strategies. Because current-generation clinical laboratories emphasize automation, flow cytometric analysis of PCR- amplified products shows increased analytic sensitivity and offers a vehicle for automation of DNA amplification tests.     

增强疏水膜半干式蛋白电泳印迹作用的实验

电泳印迹技术是蛋白质化学研究的重要手段之一 ,主要方法有垂直的槽式和水平的半干式两种 ,本实验探讨对疏水膜半干式蛋白电泳印迹的影响因素 ,旨在增强印迹作用。1　材料和方法1.1　试剂　两性载体 ( ampholine p H 3.5～ 10 .0 )购自Pharmacy公司 ;丙烯酰胺 ( Acrylamide)、N,N′-甲叉双丙烯酰胺 ( Bis)及疏水膜 ( PVDF)购自 Bio-Rad公司 ;兔抗鼠 α、β、γ抗血清由美国卫生研究院 ( NIH) Zigler JS博士惠赠 ;其余试剂均购自 Sigma公司。1.2　仪器　等电聚焦采用多用电泳仪 ( Multiphor ,L KB公司 ,瑞典 ) ;SDS-PAGE采用微…