携带TIMP-4 cDNA的重组腺病毒载体的构建

目的 克隆细胞外基质金属蛋白酶组织抑制因子4(tissue inhibitor-4 of metalloproteinases,TIMP-4)基因,构建包装携带TIMP-4基因的重组腺病毒载体(Ad/T4),为基因治疗血管再狭窄的实验研究打下基础.方法 以人心脏cDNA文库为模板,应用PCR克隆TIMP-4基因,经DNA测序确定克隆结果.构建具有强启动子CMV,携带TIMP-4和报告基因GFP的穿梭质粒pAdTrack-T4,利用AdEassy系统,在大肠杆菌BJ5183经同源重组产生重组腺病毒质粒pAd/T4.脂质体介导转染293细胞,包装重组腺病毒载体(Ad/T4).结果 克隆出含TIMP-4全部编码序列的cDNA片断,并通过亚克隆和同源重组等将其重组入腺病毒,构建包装了重组腺病毒载体Ad/T4.结论 Ad/T4中含有TIMP-4和GFP的完整表达盒,可用于基因治疗的实验.     

关于冠状动脉分叉病变角度命名的商榷

正冠状动脉左主干发出左前降支和左旋支形成分叉、前降支发出对角支形成分叉、左旋支发出钝缘支形成分叉等等,以往研究只关注斑块在分叉部位的分布~([1-5]),不重视分叉血管之间的角度。由于血管介入技术的迅速发展,分叉病变的位置直接影响介入技术的选择和预后。国内外关于分叉病变角的名称不统一(图1～3)~([6-8]),为了有效地进行学术交流和临床研究,借鉴数学关于"角"(∠)的命名原则,提出新的冠状动脉分叉角名称(图4)。命名原则如下。(1)主支与分支:冠状动脉分叉前、后血管走行一致(或接近者)者为主支血管(main vessel,MV),而另一条血管称为侧支     

孤啡肽的研究进展

１９９５年Ｍｅｕｎｉｅｒ［１］和Ｒｅｉｎｓｃｈｅｉｄ［２］等几乎同时从脑组织中分离出一种新的由１７个氨基酸组成的肽－称孤啡肽（ＯＦＱ）或痛敏肽,并证明孤啡肽特异性结合阿片受体样受体（ｏｐｉｏｉｄ－ｌｉｋｅｒｅｃｅｐｔｏｒ１ＯＬＲ１）,为ＯＬＲ１的内源性配体。由于孤啡肽前体的分子结构与阿片类前体,如强啡肽和脑啡肽原的前体分子有很高的同源性,而且ＯＦＱ分子小,有潜在的药用价值,所以一经发现就引起了人们的广泛关注,现就其分布、分子结构、基因的克隆定位以及生物学功能和药理作用等综述如下。１孤啡肽及其前体…     

NaOH消蚀法制备胎儿脱细胞真皮基质的研究

目的：探索NaOH消蚀法对胎儿真皮基质的成分及其形态结构的影响。方法：使用NaOH消蚀法制备胎儿皮肤的脱细胞真皮基质(ADM)，应用组织化学和免疫组织化学染色方法检测细胞的余留及细胞外基质成分的破坏情况，应用扫描电镜观察ADM的构筑。结果：NaOH消蚀处理16h即可去除真皮细胞成分。制备的ADM韧性好，勿需戊二醛交联。弹性纤维的含量有所减少而胶原纤维形态完整，排列正常，保留了基本的真皮构筑。HLA—DR呈阴性，Fibronectin(FN)呈弱阳性，Laminin(LN)和Ⅳ型胶原均呈阴性。结论：NaOH消蚀法简易经济，去除细胞成分较彻底，所制备的ADM韧性好，胶原纤维形态完整，排列正常，但对基膜的成分及结构破坏较重。     

脱细胞真皮基质和聚丙烯补片修补大鼠腹壁疝的实验研究

Objective To study the value of porcine acellular dermal matrix(ADM) made by complex maceration and Marlex mesh as ventral hernia-repairing material. Methods ADM was made from the full thick skin of the back of swines by complex maceration. The male SD rats were randomly divided into three groups including ventral hernia group(n =7), ADM group(the ventral hernia repaired by ADM, n =28) and Marlex mesh group (the ventral hernia repaired by Marlex mesh group, n = 10). The occurrence of ventral hernia was observed on postoperative 1 week. Six rats were respectively selected from the Marlex group and ADM group and tension resistance at 5weeks after surgery was observed. Results The occurrence of ventral hernia in ADM group and Marlex mesh group was significantly lower than that of ventral hernia group(0.04%, 0.0%, 100.0%, P ＜ 0. 01 ) in postoperative 1week, but there was no difference between ADM group and Marlex mesh group ( P ＞ 0.05 ). Marlex had a significantly higher breaking strength than ADM[(417 ±44)N vs( 111 ±27)N,P ＜0.01]. But 5 weeks after surgery, the tension resistance of ADM-fascial interface was higher than that of the Marlex-fascial [( 103 ± 27 ) N vs(71 ± 19 ) N,P ＜ 0. 05]. Conclusion As a material of repairing ventral hernia, ADM prepared by the complex maceration may be superior to Marlex mesh.     

脱细胞真皮基质和聚丙烯补片修补大鼠腹壁疝的实验研究

Objective To study the value of porcine acellular dermal matrix(ADM) made by complex maceration and Marlex mesh as ventral hernia-repairing material. Methods ADM was made from the full thick skin of the back of swines by complex maceration. The male SD rats were randomly divided into three groups including ventral hernia group(n =7), ADM group(the ventral hernia repaired by ADM, n =28) and Marlex mesh group (the ventral hernia repaired by Marlex mesh group, n = 10). The occurrence of ventral hernia was observed on postoperative 1 week. Six rats were respectively selected from the Marlex group and ADM group and tension resistance at 5weeks after surgery was observed. Results The occurrence of ventral hernia in ADM group and Marlex mesh group was significantly lower than that of ventral hernia group(0.04%, 0.0%, 100.0%, P ＜ 0. 01 ) in postoperative 1week, but there was no difference between ADM group and Marlex mesh group ( P ＞ 0.05 ). Marlex had a significantly higher breaking strength than ADM[(417 ±44)N vs( 111 ±27)N,P ＜0.01]. But 5 weeks after surgery, the tension resistance of ADM-fascial interface was higher than that of the Marlex-fascial [( 103 ± 27 ) N vs(71 ± 19 ) N,P ＜ 0. 05]. Conclusion As a material of repairing ventral hernia, ADM prepared by the complex maceration may be superior to Marlex mesh.     

SOD在肾缺血再灌注损伤大鼠肺内的表达变化

目的 观察肾缺血再灌注损伤大鼠肺内超氧化物歧化酶(superoxide dismutase,SOD)的表达变化,探讨SOD在抗氧化应激反应中的作用.方法 Wistar大鼠切除右肾,无损伤动脉夹夹闭左肾动脉,建立大鼠肾缺血再灌注损伤模型.再灌注24 h后取材(肺、肾、血).酶偶联速率法和苦味酸法分别检测血尿素氮(blood urea nitrogen,BUN)、血清肌酐(serum creatinine,SCr)浓度;HE染色法观察大鼠肾形态;硫代巴比妥酸比色法检测肺内丙二醛(malondialdehyde,MDA)的含量;采用Western blot和RT-PCR方法分别测定大鼠肺组织内抗氧化酶SOD的蛋白与mRNA表达水平.结果 HE结果显示肾缺血再灌注损伤组大鼠的肾组织受损明显.与对照组相比,肾缺血再灌注损伤组血清中的BUN和SCr、肺组织中的MDA含量均明显升高(P值均＜0.01),SOD的蛋白与mRNA的表达水平均明显升高(P值均＜0.01).结论 肾缺血再灌注损伤大鼠的肺组织发生了过氧化损伤.SOD在肾缺血再灌注损伤大鼠的肺组织内发挥了抗氧化应激的作用.     

反义凝血酶受体和p21双基因共表达对损伤的大鼠颈总动脉的影响

目的观察反义凝血酶受体(ATR)和p21双基因治疗对WKY大鼠颈总动脉损伤后新生内膜的抑制效果,探寻基因治疗再狭窄的有效途径.方法将42只雄性WKY大鼠随机分成基因治疗组(导入rAAV/AP组,n＝18)、载体对照组(导入rAAV/GFP组,n＝18)和正常对照组([WTBXn＝6).构建含有ATR和p21双基因及含有绿色荧光蛋白(GFP)基因的腺病毒伴随病毒(AAV)载体pSNAV/AP和pSNAV/GFP,用重组单纯疱疹病毒(rHSV)生产体系包装重组腺病毒伴随病毒(rAAV)rAAV/AP和rAAV/GFP.将rAAV/AP或rAAV/GFP导入拉伤的WKY大鼠的颈总动脉.分别于术后2、4、6周取材,用半定量-逆转录多聚酶链反应和免疫组织化学方法检测凝血酶受体(TR)和p21在动脉壁中的表达.图像分析和统计学检验双基因的疗效.结果逆转录多聚酶链反应和免疫组织化学结果显示外源基因已导入血管壁的新生内膜和中膜并获得稳定表达.图像和统计学分析说明基因治疗后2、4和6周,双基因治疗组(p21和ATR)的颈总动脉内膜厚度分别较载体对照组组内膜减少59.2%、58.7%、64.1%;中膜厚度分别减低7.8%、10.8%、8.3%;中膜细胞密度减低30.0%、29.4%、26.2%.结论ATR和p21双基因共表达可明显抑制损伤血管的内膜和中膜细胞的增殖.     

POTENT HYPOTENSIVE EFFECTS OF ORPHANIN FQ IN CONSCIOUS STROKE-PRONE SPONTANEOUSLY HYPERTENSIVE RATS

INTRODUCTIONOrphaninFo(OFQ)ornociceptinisanovelneuropeptidethatactivatesanopioid--likeGprotein--coupledreceptor(1).OFQboundtoitsreceptorinasaturablemannerandwithhighaffinity.Itdidnltshowanyactivityattheopioidreceptor(2).NorthernanalysisofhumanandrattissuesshowedthattheOFQprecusormRNAismainlyexpressedinthecentralnervoussystem(brainandspinalcord)and,albeitweakly,intheovary.Byusingaradiationhybridcelllinepanel,theOFQprecusorgenewasmappedtotheshortarmofhumanchromosome8(spZI)(3,4).Nort…