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.     

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.     

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。     

紫杉醇联合顺铂与足叶乙甙联合顺铂方案治疗非小细胞肺癌

目的观察比较紫杉醇联合顺铂（PT）与足叶乙甙联合顺铂（PE）方案治疗非小细胞肺癌（NSCLC）的近期疗效和毒副作用。方法将77例NSCLC患者随机分为胛组和PE组,化疗2周期后比较近期疗效和毒副作用。结果PT组完全缓解（CR）1例,部分缓解（PR）16例;PE组CR 0例,PR 11例。总有效率盯组43．6％,PE组28．9％,初治有效率PT组50．0％,PE组31．0％;腺癌有效率门组47．4％,PE组23．5％,两组比较差异有统计学意义（P〈0．05）,主要毒副作用为白细胞下降,Ⅲ～Ⅳ度骨髓抑制发生率PT组15．4％,PE组13．2％;Ⅲ～Ⅳ度消化道反应两组相似。结论町方案治疗NSCLC近期疗效较PE方案明显,毒副作用两组相似。     

试论胆石症的辨治

胆石症是临床常见疾病之一，祖国医学虽无此类病名，但根据其临床表现多属“胁痛”、“黄疸”等范畴。１病因病理 胆石症的形成，主要是由于情志不畅，寒温不适，饮食不节或虫积等原因，导致肝胆功能失调。由于肝气郁结，疏泄不利而影响于胆，使胆气失于通降，胆汁郁积。肝胆气郁，湿热内蕴，煎熬胆汁，日久而成砂石。如果湿从火化，热毒炽盛而见高热寒颤；瘀热在里，胆热液溢则发为黄疸；肝胆气郁，失于疏泄，则右上腹绞痛彻背。本病病位在肝胆，若胆失通降，可累及脾胃。反之，若胃失和降，气机升降失调，亦必然影响胆的通降，使胆气郁结…