Preparation of gfp-bcl-XL-contained recombinant adenovirus vector by the homologous recombination in bacteria

AIM: To prepare gfp-bcl-X_L-contained recombinant adenovirus(rAd-gfp-bcl-X_L).METHODS: Bcl-X_L gene was amplified from pEGFP-C₃-bcl-X_L, subcloned into shuttle plasmid and formed transfer plasmid of pAdTrack-CMV-bcl-X_L. Then pAdTrack-CMV-bcl-X_L was linealinzed with PmeI and co-transformed into BJ5183 bacteria with adenovirus genomic plasmid of pAdEasy-1. The identified recombinant adenovirus plasmid was digested with PacI and transfected into 293 cells to package recombinant adenovirus particles. The target gene was detected by PCR.RESULTS: There were about 35% positive recombinant bacterial clones after the co-transformation of pAdTrack-CMV-bcl-X_L and pAdEasy-1 into BJ5183. Recombinant adenovirus particle were produced and further amplified after the transfection of pAdEasy-1-gfp-bcl-X_L into 293 cells. PCR test indicated that the recombinant Ad contained bcl-X_L gene. The titer of the purified rAd-gfp-bcl-X_L was 6.5×10¹² PFU/L. CONCLUSIONS: The homologous recombination in bacteria is a convenient and high efficient method to prepare rAd-gfp-bcl-X_L. This affords a good gene transfer vector for the gene therapy in human’s diseases.     

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.     

Effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts

AIM: To investigate the effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts.METHODS: hMSC were separated from human marrow and expanded in cuture medium. hMSC were induced with dexamethasone, β-glycerophosphate, vitamin C which acted as osteoblast differentiation inducer. PD98059 was added into the osteoblasts induction medium. The cells were assayed with cell morphology, alkaline phosphatase (AP) activity and calcium deposition. RESULTS: The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. After induced with osteoblasts induction medium, the cells showed changes in cell morphology from spindle-shape to cuboidal and polygonal. The AP activity increased gradually and reached the peak in 12 days, then decreased. Many scattered tangerice calcium nodes were observed. PD 98059 significantly inhibited AP activity and calcium deposition in a dose-dependent manner. A striking observation of the present study was that a few adipocytes appeared in cultures that were treated with PD 98059 and osteogenic differentiation medium. CONCLUSION: These results indicated that osteogenic diferentiation from the hMSCs was related to the activation of the ERK.     

Relationship between expression of somatostatin and DNA ploidy in breast cancer

AIM: To investigate the relationship between somatostatin and the pathologic type, estrogen receptor,DNA ploidy of nuclei in tumor cells of breast cancer.METHODS: 67 cases of primary breast cancer and 25 cases of benign breast tumor were examined by immunohistochemical stretomyces avidin peroxidase method. 26 cases of breast cancer selected at random were analyzed by flow cytometry.RESULTS: Somatostatin expressed significantly higher in low malignant breast cancer than that in high malignant breast cancer (P<0.05). Most of cancers with positive staining of somatostatin were diploidy,most of cancers with negative staining were aneuploidy,there had significant difference between two groups (P<0.05).CONCLUSION: Somatostatin may delay the progress of breast cancer,and somatostatin levels in cancer tissues may become a useful indicator for assessing prognosis of patients with breast cancer.     

蔗渣滤泥培养料栽培平菇试验

以不同比例滤泥代替栽培料中的麦麸进行平菇栽培试验。结果表明 ,滤泥能替代栽培料中的麦麸。     

利用RT-PCR与PVP-ELISA检测水稻瘤矮病单头叶蝉

 目前国内外对水稻瘤矮病毒(Rice gall dwarf virus,RGDV)的研究主要集中在其核衣壳蛋白和外衣壳蛋白的性质、结构及其与介体的亲和性、与介体传毒能力的关系等方面,本室在已有室内检测研究的基础上,利用RT-PCR和PVP-ELISA法对田间带毒叶蝉进行检测,并对2种方法的检测结果进行了比较。     

喀喇昆仑山盖孜河水文水资源特性分析

喀喇昆仑山河流自有水文资料记载以来 ,近 10年洪灾发生频率明显高于以往 ,人民的生命财产损失程度也大幅度增加。在分析了盖孜河流域的径流及气候多年变化过程后 ,认为洪灾频发的原因是近年来流域内降水及气温呈上升趋势 ,导致高山冰川退缩 ,河流径流增加 ,洪灾增大、频率增高 ;小区域多年气候变化过程从侧面印证了我国西北地区气候由暖干向暖湿变化。     

晋西昕水河流域生态经济型防护林体系景观格局动态分析

本文以晋西昕水河流域为典型实例，概述了流域生态经济型防护林体系建设背景。依据流域内各类型区在不同时段的土地利用结构和生态经济特征，进行流域生态经济型防护林体系景观格局变化分析，表明了流域内景观多样性和异质性不断增加的发展趋势，以及确定各区主要发展方向，实行综合开发与治理的必要性，并提出了相应的流域生态经济型防护林体系动态调控途径。     

奥芬达唑和阿苯达唑对猪囊尾蚴作用形态学比较观察

观察了奥芬达唑和阿苯达唑对猪体内及体外培养的不同发育阶段囊尾蚴作用的形态学效果.结果表明奥芬达唑具有显著的疗效,优于阿苯达唑,且对未成熟期猪囊尾蚴的杀灭作用优于成熟期.提示奥芬达唑可能成为抗未成熟期猪囊尾蚴及治疗脑囊虫病的有效药物.     

以甜菜粕取代小麦的全混合颗粒饲料对羔羊瘤胃pH值的影响

本试验选用24只3-3.5月龄的波德代(♂)×蒙古羊(♀)和陶赛特(♂6)×蒙古羊(♀)杂交一代羔羊,按2×2×2因子设计,研究了两个营养水平下以高果胶的甜菜粕取代小麦的全混合颗粒饲料对两种羔羊瘤胃pH值的影响。结果表明,低营养水平组的pH值显著高于高营养水平组(P=0.015);食后4 h(P-0.019)、6 h(P=0.009)的pH值在营养水平和饲料间存在互作,高营养水平组,小麦组羔羊的瘤胃液pH高于甜菜粕组,而低营养水平组、甜菜粕组的pH高于小麦组;羊品种对pH值无影响。