利用RNA干扰技术治疗慢性心衰大鼠的研究

目的利用RNA干扰（RNA interference，RNAi）技术抑制心衰大鼠受磷蛋白（phospho—lamban，PLN）mRNA的表达，改善心脏功能。方法构建表达PLN shRNA的重组腺相关病毒载体（rAAV2-phRi1及rAAV2-phRi2）；通过经腹心包穿刺术将rAAV2-phRi1及rAAV2-phRi2注入心衰大鼠心包内，基因导入后10d，30dRT—PCR法检测RNAi对心肌细胞PLN mRNA的抑制效应，并测定肌浆网Ca^2＋-ATP酶（sarcoplasmic reticulum Ca^2＋-ATPase，SERCA2a）活性，比较血流动力学的变化。结果rAAV2-phRi1及rAAV2-phRi2心包注射后，与心衰组比较，10d时PLN mRNA分别降低18．76％／16．36％，SERCA2a活性及血流动力学指标无明显改善；30d时rAAV2-phRi1及rAAV2-phRi2对PLN mRNA的抑制效率为85．32％／67．65％，SERCA2a活性增加104％／74％，血流动力学显著改善（P〈0．05），达到假手术组水平（P〉0．05）；rAAV2-phRi1的抑制效率，对心功能的改善显著优于rAAV2-phRi2（P〈0．05）。结论在体水平上，RNAi能够抑制心衰大鼠PLN mRNA表达，改善心脏功能，rAAV2-phRi1优于rAAV2-phRi2。     

鸭乙肝病毒的聚合白蛋白受体

为研究DHBV的宿主限制性机制,探讨基因组Pre-S编码的生物学意义。本文用PDSA,PHSA PBSA分别包被羊红细胞,检测了140启东鸭、20只广西扶绥鸭和20只长春鸭的聚合白蛋白结合活性,同时做了血清和肝组织中DHBV、DNA斑点杂交和组织学检查。83/140的启东鸭、5/20扶绥鸭、1/18长春鸭的PDSA结合活性增高,5/140启东鸭、1/20扶绥鸭和1/18长春鸭的PHSA和PBSA     

用分子杂交方法研究鸭乙肝病毒感染和鸭肝病

嗜肝 DNA 病毒感染同肝炎、肝硬化和肝癌的相关关系是肝病学中的重大课题。除了 HBV 感染同人类肝病和肝癌的发生有关之外,一些动物嗜肝 DNA 病毒也有类似性。尽管 WHV、GSHV 和 DHBV 有类似于 HBV的形态、结构以及都能造成长期持续性感染,但 WHV 感染的土拔鼠和 GSHV 感染的金花鼠,两者的肝病病理学改变有显著差     

肝病患者的糖耐量、胰岛素和C肽

417例急性肝炎、慢性肝炎、肝硬变和原发性肝癌病人的葡萄糖耐量试验表明,58.9%急性肝炎,69.4%慢性肝炎,72%肝硬变和72.7%原发性肝癌病人有异常的糖耐量曲线。其中49.3%急性肝炎为低反应型,41.5%慢性肝炎和60.6%肝硬变病人为高反应型。血清胰岛素测定表明,慢性肝炎组空腹胰岛素为0.143±0.056pmol/ml,肝硬变组0.203±0.076pmol/ml,明显高于对照组,     

Anticancer effect of icaritin on human lung cancer cells through inducing s phase cell cycle arrest and apoptosis

Icaritin, a prenylflavonoid derivative from Epimedium Genus, has been shown to exhibit many pharmacological and biological activities. However, the function and the underlying mechanisms of icaritin in human non-small cell lung cancer have not been fully elucidated. The purpose of this study was to investigate the anticancer effects of icaritin on A549 cells and explore the underlying molecular mechanism. The cell viability after icaritin treatment was tested by MTT assay. The cell cycle distribution, apoptosis and reactive oxygen species（ROS） levels were analyzed by flow cytometry. The mRNA and protein expression levels of the genes involved in proliferation and apoptosis were respectively detected by RT-PCR and Western blotting. The results demonstrated that icaritin induced cell cycle arrest at S phase, and down-regulated the expression levels of S regulatory proteins such as Cyclin A and CDK2. Icaritin also induced cell apoptosis characterized by positive Hoechst 33258 staining, accumulation of the Annexin V-positive cells, increased ROS level and alteration in Bcl-2 family proteins expression. Moreover, icaritin induced sustained phosphorylation of ERK and p38 MAPK. These findings suggested that icaritin might be a new potent inhibitor by inducing S phase arrest and apoptosis in human lung carcinoma A549 cells.     

Fucoidan induces apoptosis of HepG2 cells by down-regulating p-Stat3

Summary： Fucoidan is one of the main bioactive components of polysaccharides. The current study was focused on the anti-tumor effects of fucoidan on human heptoma cell line HepG2 and the possible mechanisms. Fucoidan treatment resulted in cell cycle arrest and apoptosis of HepG2 cells in a dose-dependent manner detected by MTT assay, flow cytometry and fluorescent microscopy. The results of flow cytometric analysis revealed that fucoidan induced G2/M arrest in the cell cycle progression. Hoechst 33258 and Annexin V/PI staining results showed that the apoptotic cell number was increased, which was associated with a dose-dependent up-regulation of Bax and down-regulation of Bcl-2 and p-Stat3. In parallel, the up-regulation of p53 and the increase in reactive oxygen species were also observed, Which may play important roles in the inhibition of HepG2 growth by fucoidan. In the meantime, Cyelin B 1 and CDK1 were down-regulated by fucoidan treatment. Down-regulation of p-Stat3 by fucoidan resulted in apoptosis and an increase in ROS in response to fucoidan exposure. We therefore concluded that fucoidan induces apoptosis through the down-regulation of p-Stat3. These results suggest that fucoidan may be used as a novel anti-cancer agent for hepatocarcinoma.     

氧(酸)化电位水杀灭微生物及对金属腐蚀性实验研究

氧（酸）化电位水在２０°Ｃ条件下，对大肠杆菌（ＡＴＣＣ８０９９）、金黄色葡萄球菌（ＡＴＣＣ６５３８）作用１分钟杀灭率可达１００％；对枯草杆菌黑色变种（ＡＴＣＣ９３７２）芽胞作用３分钟平均杀灭率达９９．９４％，作用５分钟以上，平均杀灭率可达９９．９９％以上；对乙肝表面抗原（ＨＢｓＡｇ）作用３０秒种能使其转阴。有机物存在能明显降低杀菌效果，对铝、铜、碳钢金属呈中度腐蚀。     

通用腺相关病毒载体构建及表达报告基因GFP的研究

目的　构建基因治疗通用型 AAV载体并检测它转导外源基因作用。方法　使用限制性内切酶切出p SSV9int-质粒中的 AAV病毒 Rep和 Cap基因元件后 ,插入了重组腺病毒专用穿梭质粒 -p ACCMVp L p A的含有CMV启动子、多克隆位点 (MCS)和多聚腺苷酸信号 (Poly A)的表达盒 ,构建了重组 AAV通用载体质粒 p SSHG-CMV。在该质粒 MCS插入 GFP基因后 ,我们使用 p SSHG-CMV-GFP、p GF14 0和 p AAV/Ad 3种质粒共转染 2 93包装细胞 ,制备 GFP重组 AAV,应用斑点杂交实验检测重组病毒滴度度 ,并将该病毒感染新生大鼠星形胶质细胞 ,荧光显微镜观察 GFP表达。结果　重组 AAV的滴度在浓缩前可达 2× 10 11,浓缩后可达 2× 10 13 ,表明成功的构建了重组 AAV载体 ,插入外源基因 GFP后 ,在包装病毒和辅助质粒的联合作用下 ,能产生具有感染性的重组AAV。感染了重组 GFP-AAV的大鼠星形胶质细胞表达了明显的 GFP荧光。结论　本文构建的重组 AAV通用载体 p SSHG-CMV,可转导外源基因 ,用于基因治疗的研究     

腹部皮管治疗手指套状皮肤缺损31例临床报道

目的探讨腹部皮管修复手指套状皮肤缺损的治疗价值。方法2003年1月-2005年6月，笔者采用改良的腹部皮管的方法治疗手指套状皮肤缺损病人31例共31指，3个月到1年，平均9个月。Ⅰ型：远侧指间关节以远皮肤缺损；Ⅱ型：近侧指间关节以远皮肤缺损；Ⅲ型：掌指关节以远皮肤缺损。Ⅰ型、Ⅱ型病例去除末节指骨1—2mm。Ⅲ型病例去除末节指骨，保留2节指骨。细克氏针固定指间关节伸直位。于同侧下腹部设计任意腹部皮辨。皮辨远1／3只保留真皮下毛细血管网，中1／3只带小量脂肪组织，蒂部1／3带有较多脂肪组织及少量筋膜层。3周开始皮管血运训练，6周断蒂。断蒂时，原切口切开皮管再次修簿，采用5—0无创伤缝合线，将掌侧真皮组织与深部组织缝合。结果所有病例皮瓣血运良好，手指外观良好。Ⅰ型、Ⅱ型病例保留了手指原来长度，Ⅲ型病例保留了2节手指长度，经过功能锻炼伤指指间关节也有不同程度恢复。结论采用改良的腹部皮管的方法治疗手指套状皮肤缺损，尽量保留了手指长度，外观上接近原来的手指；并且保留了伤指关节的部分屈伸活动；手指掌侧的皮肤滑动明显减少，有利于抓握，持物。此治疗方法疗效可靠，有广阔的应用价值。     

先天性肛门闭锁伴结肠易位一例报告

<正> 先天性消化管畸形中最多见的是肛门闭锁。高位和中位肛门闭锁常伴有肠道泌尿生殖道瘘,其中少数病人同时伴有短结肠畸形和膀胱外翻,被称为“内脏外翻”或“先天性肛门闭锁伴发短结肠畸形”。先天性肛门闭锁伴发结肠易位未见报告,近来我们遇到一例,报告如下:病历摘要吕××,男,64岁,退休工人,因肝硬变,腹水于