Construction and identification of recombinant adenovirus vector containing CTLA4Ig gene

AIM:To construct a recombinant adenovirus expression vector containing CTLA4Ig gene.METHODS:The CTLA4Ig gene derived from the plasmid PCDNA3.0/CTLA4Ig by using polymerase chain reaction (PCR) was inserted into the backward position of cytomegalovirus (CMV) immediate early promoter of the shuttle plasmid (pAdTrack-CMV). After being identified by endonuclease, PCR and sequencing, the recombinant shuttle plasmid pAdTrack-CTLA4Ig was co-transformed into E.coli. BJ5183 cells with the adeoviral backbone plasmid pAdEasyl-1 to obtain the homologous recombination. The adenovirus was generated in 293 cells. A series methods such as PCR and fluorescence microscope was employed to identify the generated recombinant adenovirus.RESULTS:Recombinant CTLA4Ig adenoviruses were constructed and the titer of virus was generally up to 1.65×10¹² phaque forming units per liter (PFU/L).CONCLUSION:Success in constructing recombinant pAdTrack-CTLA4Ig will be the base of the further research on its expression in the mammalian cells, and be potenially used in the prevention of transplant rejection and autoimmunity diseases.     

Selection and amplification of the liver stem cell subset from rat bone marrow cells with a medium containing cholestatic serum in vitro

AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro. METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.     

篱壁式红富士苹果光照分布及结果部位对果实品质的影响

作者对6～7年生篱壁式红富士苹果叶幕光照分布、结果部位对品质的影响进行了研究.结果表明:篱壁形叶幕光照垂直分布和水平分布的日变化相对均衡,树冠内未产生无效光区;品质垂直和水平分布与光照垂直和水平分布呈极显著正相关.中、下层中长果枝果实的果面光洁度分别为92.7%和97.2%,端正果率分别为91.6%和92.5%.     

Expression of alpha-smooth muscle actin in scar tissue and its relationship with apoptosis

AIM: To examine the expression of alpha-smooth muscle actin in scar tissue, and observe the phenomenon of apoptosis and its involvement in the process of pathological scarring and the presence of myofibroblasts or absence of cell in the dermis. To investigate the potential role of reparative cell apoptosis in hyperplastic scar formation. METHODS: The samples of scar were obtained from post-burn patients undergoing plastic operation in our burn unit recently, and the samples of control came from skin donor site of the same patient correspondingly. TUNEL assays were performed to evaluate the number of apoptotic cells in scar versus normal skin. In situ hybridization and immunohistochemistry staining technique were employed to determine the expression of different dermis cells markers in scar tissue and normal skin. RESULTS: There existed evident difference in apoptotic cells in the dermis between scars tissue and normal human skin. The expression positive cells were much more in hyperplastic scars than that in normal human skin; the apoptotic cells of proliferative stage were slight more than that of mature stage. However, in proliferative stage, the number of apoptotic cells was higher for the combination of hyperplastic scar than normally healed flat scars. But in mature stage, no obviously difference was detected between hyperplastic scar and normally healed flat scar. The monoclonal anti-α smooth muscle actin (ASMA) expression was significantly stronger in proliferative stage than that of mature stage. CONCLUSIONS: With reconstitution of dermal tissue, myofibroblasts containing alpha-SM actin disappear under normal wound healing, probably as a result of apoptosis. The myofibroblast play a critical role in wound closure and in the pathologic sequelae of healing.     

The genetic susceptibility of HLA-DRB1 alleles in esophageal neoplasm of Hubei Han Chinese

AIM: To probe into the genetic susceptibility of HLA-DRB1 alleles to esophageal neoplasm in Hubei Han Chinese. METHODS: HLA-DRB1 gene polymorphism in 42 patients with esophageal neoplasm and 136 normal control subjects was studied by PCR and sequence. RESULTS: Allele frequency of HLA-DRB1 *0901 allele was significantly higher in esophageal cancer patients than those in normal controls(0.2500 vs 0.1397, P =0.028; the odds ratiO².053; etiologic fraction 0.1282).There were no association between the rested HLA-DRB1 alleles with patients. CONCLUSION: Individuals carrying HLA-DRB1 *0901 may be susceptible to esophagealo carcinoma, its nucleotide sepuence approachs to the corresponded allele sequence(exoN²)published in GenBank.     

达县分葱田甜菜夜蛾发生规律初步研究

采用系统调查方法研究了达县地区分葱田甜菜夜蛾的生活史、发生与环境的关系。甜菜夜蛾在达县分葱田1a发生6代,以三至五代为害最重,世代重叠。其数量消长与虫源、夏秋季气候、天敌、栽培管理等因素密切相关。通过调查分析提出本县初始虫源非本地虫源。     

红汁乳菇多糖提取最佳工艺研究

对红汁乳菇多糖提取、纯化工艺进行了优化研究，结果表明在料液比1：30，浸提温度95％浸提时间2h,pH8．0，可获得最佳提取效果。在醇析浓度为70％，V(氯仿)：V(正丁醇)=1：0．2，V(氯仿 正丁醇)：V(样品)=1：l时，纯化含量为58％。同时用超声波法对红汁乳菇多糖的提取作了研究，其得率为6.99％，效果较佳。     

鸡腿蘑菌丝体的同工酶研究

用8种不同培养基配方，培养鸡腿蘑菌丝体，其同工酶分析结果表明，鸡腿蘑菌丝体酯酶(EST)同工酶谱差异明显，过氧化物酶(POD)同工酶谱差异明显。     

鸡球虫对机体的致病作用研究进展

球虫在鸡体内进行生命活动的同时对鸡体产生不同程度的损害，球虫感染后的致病性和病理学反应是球虫与宿主机体相互作用的结果。本文就鸡球虫致病作用及其对宿主细胞代谢的影响、及影响球虫致病性的因素的研究进展予以综述，对鸡球虫病的防治具有一定的指导意义。     

丝兰属植物提取物降低鸡舍氨气浓度的效果试验

通过在21日龄的肉用仔鸡饲料中添加丝兰属植物提取物(其有效成分为丝兰皂角苷)，研究丝兰属植物提取物对降低鸡舍中氨气浓度和提高肉仔鸡生产性能的效果。试验结果表明，试验组鸡舍内氨气平均浓度为4．75mg／L，对照组鸡舍内氨气平均浓度为13．80mgl／L，试验组比对照组鸡舍内氨气平均浓度降低了9．05mg／L，经t检验，两鸡舍内氨气平均浓度差异显著(P＜0．05)，并且试验组比对照组饲料报酬提高了8.92％。