免疫组化P16和ki67双表达在宫颈癌前病变及宫颈癌的临床分析

目的 分析在宫颈癌前病变及宫颈癌临床诊断时采取免疫组化P16和ki67双表达方法的临床价值。方法 选取2016年1月—2021年12月宿迁市第一人民医院收治的70例宫颈癌前病变患者作为对照组,另选择同期收治的50例宫颈癌患者作为观察组。两组均以相同的方式检测其病理组织标本中的P16、ki67表达水平,并分析P16、ki67单一检测及联合检测的诊断效能。结果 观察组P16表达量(3.03±0.24)分、ki67表达量(2.11±0.32)分均显著高于对照组,差异有统计学意义(t=75.379、41.008,P<0.05);联合检测真阳性检出率、敏感度、特异度、准确性均显著高于P16单一检测、ki67单一检测,差异有统计学意义(χ²=8.305、7.161、17.472、7.600、11.002,P<0.05)。结论 与宫颈癌前病变患者P16、ki67表达水平相比,宫颈癌患者主要为高表达,联合检测具有更高的诊断效能,因而在临床中的使用价值突出,适合普及。     

宿迁地区女性人群中HPV病毒感染情况及亚型分析

目的 了解宿迁地区人乳头瘤病毒(human papilloma virus,HPV)分型感染情况,分析当地HPV分布特点及规律,为宿迁地区宫颈病变地防治提供可靠依据。方法 选择2019年1月—2020年1月在宿迁市第一人民医院就医并行HPV分型检测的女性患者共6 978例,从年龄、HPV亚型分布及单一、混合感染情况,分析宿迁地区HPV感染分布特征。结果 在6 978例样本中,HPV阳性1 034例,感染率为14.82%,高危型HPV感染占67.92%,疑似高危型感染占10.45%,低危型HPV感染占21.63%。其中单一感染389例(37.62%),多重感染645例(62.38%),多重感染中二重感染最为常见,共397例(61.55%)。感染率排在前5位的HPV亚型为52、16、58、53、39型,感染率最高的两组为<30岁(17.24%)及≥60岁(18.75%),除<30岁年龄段外,随着年龄的增加,HPV高危型感染阳性率呈现逐渐升高的趋势;且随着年龄的增加,三重感染比例升高(χ²=12.603,P<0.05)。结论 宿迁地区HPV感染以多重感...     

大鼠肝细胞生长因子与肝再生增强因子融合基因真核表达质粒的构建及鉴定

Objective To construct and identify an eukaryotic expression plasmid containing rat hepatocyte growth factor(rHGF)gene and rat augmenter of liver regeneration(rALR)gene,so that to provide experimental basis for developing new treatments of hepatic fibrosis.Methods The gene fragments of rHGF and rALR were amplified from recombinant prokaryotic plasmid pUC18-rHGF and pUC18-rALR by polymerase chain reaction(PCR),respectively,then were spliced by overlap extension PCR with a linker,and the fusion gene rHGF-linker-rALR was constructed.The fusion gene was directionally inserted into the eukaryotic expression plasmid pcDNA3.1 between restriction sites of Kpn Ⅰ and Xba Ⅰ to construct the recombinant eukaryotic expression plasmid pcDNA3.1-rHGF-linker-rALR,and the new constructed recombinant plasmid was identified by double restriction digestion and DNA sequencing.Results DNA fragments of 2200 bp and 400 bp were observed after the electrophoresis of products amplified from recombinant prokaryotic plasmid pUC18-rHGF and pUC18-rALR,respectively,which was consistent with the theoretical value.The electrophoresis of fusion gene rHGF-linker-rALR obtained by overlap extension PCR technique showed only a 2 600 bp DNA fragment,which was in accordance with the expected value.Electrophoresis of products of pcDNA3.1-rHGF-linker-rALR digested with Kpn Ⅰ and Xba Ⅰ showed two DNA fragments with 2600 bp and 5400 bp,which were both consistent with the expected value.The sequences were confirmed correctly by DNA sequencing.Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1-rHGF-linker-rALR is successfully constructed,which provides experimental basis for developing gene therapy of hepatic fibrosis.     

大鼠肝细胞生长因子与肝再生增强因子融合基因真核表达质粒的构建及鉴定

Objective To construct and identify an eukaryotic expression plasmid containing rat hepatocyte growth factor(rHGF)gene and rat augmenter of liver regeneration(rALR)gene,so that to provide experimental basis for developing new treatments of hepatic fibrosis.Methods The gene fragments of rHGF and rALR were amplified from recombinant prokaryotic plasmid pUC18-rHGF and pUC18-rALR by polymerase chain reaction(PCR),respectively,then were spliced by overlap extension PCR with a linker,and the fusion gene rHGF-linker-rALR was constructed.The fusion gene was directionally inserted into the eukaryotic expression plasmid pcDNA3.1 between restriction sites of Kpn Ⅰ and Xba Ⅰ to construct the recombinant eukaryotic expression plasmid pcDNA3.1-rHGF-linker-rALR,and the new constructed recombinant plasmid was identified by double restriction digestion and DNA sequencing.Results DNA fragments of 2200 bp and 400 bp were observed after the electrophoresis of products amplified from recombinant prokaryotic plasmid pUC18-rHGF and pUC18-rALR,respectively,which was consistent with the theoretical value.The electrophoresis of fusion gene rHGF-linker-rALR obtained by overlap extension PCR technique showed only a 2 600 bp DNA fragment,which was in accordance with the expected value.Electrophoresis of products of pcDNA3.1-rHGF-linker-rALR digested with Kpn Ⅰ and Xba Ⅰ showed two DNA fragments with 2600 bp and 5400 bp,which were both consistent with the expected value.The sequences were confirmed correctly by DNA sequencing.Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1-rHGF-linker-rALR is successfully constructed,which provides experimental basis for developing gene therapy of hepatic fibrosis.     

高危组肾移植患者血浆置换后进行HLA配型1例

患者,男性,43岁,既往诊断为原发性肾小球肾炎所致尿毒症.2005年1月肾移植术后,因为发生超急性排斥而导致移植肾丧失功能,患者血液透析维持治疗并欲进行第二次肾移植.     

肝细胞生长因子与肝再生增强因子重组表达质粒对大鼠肝纤维化的影响

Objective To evaluate the therapeutic effects of recombinant expression plasmid containing hepatocyte growth factor (HGF) and augmenter of liver regeneration (ALR) on rats with hepatic fibrosis. Methods Ninety Sprague-Dawley rats, which had been established into hepatic fibrosis models, were equally divided into 6 groups: blank group, pcDNA3.1 therapy group,pcDNA3.1-HGF therapy group, pcDNA3. 1-ALR therapy group, pcDNA3.1-HGF and pcDNA3. 1-ALR combined therapy group, and pcDNA3. 1-HGF-ALR therapy group. Zero point one μmol of blank or plasmid was injected into model rats in each group by tail vein once a day for 3 days. Model rats in blank group didn't receive any treatment. Additional 10 rats were chosen as control group, which were not given any interference during the experiment. All rats were sacrificed 4 days after end of treatment. Liver tissues were reserved for observing pathologic changes after HE staining and detecting proliferating cell nuclear antigen (PCNA) and c-jun by immunohistochemistry. Measurement data were compared by single-factor analysis of variance. Comparison between groups was done by SNK test. Enumeration data were analyzed by Fisher's exact test. Results In blank group and pcDNA3.1 therapy group, hyperplasia of fibrous connective tissue was very obvious, false lobules were formed. There was no significant difference between these two groups (x2 =0. 317,P= 1. 000).In the 4 remaining groups, hepatic fibrosis all achieved different degree of amelioration, and the therapeutic effect of pcDNA3.1-HGF-ALR was optimal. In control group, the expressions of PCNA and c-jun in liver tissues were low, with absorbance value of 8.6±1.9 and 3.2 ± 1.2, respectively. In blank group and pcDNA3. 1 therapy group, the expressions of PCNA and c-jun were obviously increased, with absorbance value of 24. 1±3.0, 24.5±4.3 and 23.8±3.1, 24.9±4.2, respectively,which were significant different from control group (all P＜0.01). In the 4 remaining groups, the expressions of PCNA were all obviously increased, and expressions of c-jun were all obviously decreased. The maximum change scope was observed in pcDNA3. 1-HGF-ALR therapy group.Conclusions The recombinant expression plasmid pcDNA3. 1-HGF-ALR can effectively ameliorate experimental hepatic fibrosis of rats. The anti-fibrosis effects are achieved probably by up-regulating PCNA expression and down-regulating c-jun expression.     

盲袢瘘的防治体会

任何一种新改进的术式都应在长期大量的实践中考证,才能得出正确的评价。作者的实事求是的态度对盲袢式胆管空肠吻合术并发盲袢瘘的问题作了较详细的总结,很有意义。本文提示:①任何手术均应严格掌握其适应证,取利避害。②本术式是胆肠内引流术的一种,它也只能在有效地解除肝内梗阻,去除肝内病灶的前提下应用,不能把解决肝内病变的努力,留待术后经盲袢的有限处理上。③一旦发生盲袢并发症,病人的负担和处理的困难都是不小的。