par-4 SAC基因的克隆及序列测定

目的:利用基因工程技术对 par-4 SAC 进行基因克隆并测定其序列,为进一步诱导肿瘤细胞靶向性凋亡的基因治疗奠定基础.方法:采用非对称互补引物/模板法,制备两端含有酶切位点的 par-4 SAC 的 cDNA,PCR产物克隆入 pGEM-T Easy 载体并转化感受态大肠杆菌 E.coli DH5α菌株,随机挑取数个菌落,筛选鉴定并测序.结果:经酶切鉴定、测序分析,表明所插入的基因片断为 par-4 SAC 基因,与设计完全相同.结论:应用非对称互补引物/模板法克隆 par-4 SAC 基因是成功及可行的.     

肝性腹水的发生机制

腹水是肝硬化病人常见而突出的临床体征。顽固性腹水不但治疗困难,而且提示病人预后不佳。肝性腹水可以自发或由感染、手术、上消化道大出血等促发而成,但腹水的形成和发展都是由全身病理生理变化和门静脉、肝静脉、肝淋巴液回流障碍两种因素决定的。充分地认识肝性腹水形成的有关     

par-4 SAC基因的克隆及序列测定

目的：利用基因工程技术对par-4 SAC进行基因克隆并测定其序列，为进一步诱导肿瘤细胞靶向性凋亡的基因治疗奠定基础。方法：采用非对称互补引物／模板法，制备两端含有酶切位点的par-4 SAC的cDNA，PCR产物克隆人pGEM—T Easy载体并转化感受态大肠杆菌E．coli DH5α菌株，随机挑取数个菌落，筛选鉴定并测序。结果：经酶切鉴定、测序分析，表明所插入的基因片断为par-4 SAC基因，与设计完全相同。结论：应用非对称互补引物／模板法克隆par-4 SAC基因是成功及可行的。     

可分泌表达神经保护肽的重组慢病毒对闭合性脑损伤小鼠的神经保护作用

目的：通过滴鼻给药途径,给予闭合性脑损伤小鼠可分泌表达神经保护肽NAP的重组慢病毒,观察该重组病毒经鼻-脑通路对中枢神经系统的保护作用,为携带可分泌表达NAP的重组慢病毒治疗神经系统退行性疾病提供理论依据。 方法：选用8~12周成年雄性昆明小鼠54只,随机分为4组：空白对照组10只（Control组）、重锤加害组20只（CHI组）、重组慢病毒rLent/NT4-NAP保护组20只（rLent组）和重组慢病毒rLent/GFP组4只（GFP组）。观察各组小鼠的死亡率、神经功能损伤（NSS）评分、脑水肿含量和病理改变情况。应用共聚焦显微镜观测GFP组小鼠鼻黏膜、嗅神经和脑组织内绿色荧光表达情况。 结果：rLent组小鼠死亡率（15％）明显低于CHI组小鼠（55％）;rLent组小鼠NSS评分在创伤后1、3、5和7 d均显著低于CHI组（P<0.01）; rLent组小鼠创伤后24 h的脑水肿含量明显低于CHI组（P<0.01）;HE染色显示rLent组小鼠病理改变明显轻于CHI组;GFP组仅在鼻黏膜处可见呈条索样的绿色荧光,而在嗅神经及大脑则未发现绿色荧光。 结论：分泌表达NAP的重组慢病毒可感染鼻黏膜细胞,分泌表达的短肽NAP能够改善闭合性脑损伤小鼠的神经功能;未加装分泌表达元件的重组病毒rLent/GFP仅能感染鼻黏膜细胞,不能通过血脑屏障进入中枢神经系统。     

陕西省莱姆病血清流行病学调查研究

目的:调查了解莱姆病在陕西省人群中的自然感染情况。方法:间接免疫荧光(IFA)检测方法和现场流行病学调查。结果:通过血清流行病学调查,证实三个调查地区人群中均有莱姆病的自然感染,阳性率分别为8.33%(6/113)、8.88%(3/36)和7.51%(4/45);职业分布是以林业工人居首位(阳性率10.6%),山区农民次之(阳性率6.57%),林业机关干部居第三位(阳性率4.0%),林场家属和山区教师本次未检出阳性结果;年龄与性别分布是以青壮年男性人群自然感染率较高。结论:证实陕西省部分林区和山区人群中有莱姆病存在及其自然感染,为防治提供了科学依据。     

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.     

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.     

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