Effects of TGF-β1 and TNF-α antisense PS-ODNS on ex vivo expansion of hematopoietic stem/progenitor cells(HSPC)

AIM:To study the effect of TGF-β₁ and TNF-α antisense PS-ODNS on ex vivo expansion of hematopoietic stem/progenitor cells (HSPC). METHODS:CD₃₄⁺cells were purified from fresh umbilical cord blood by immunomagnetic beads, and mononuclear cells were purified from bone marrow by Ficoll-hypaque. The effects of TGF-β₁ and /or TNF-α antisense PS-ODNS on ex vivo expansion of CD₃₄⁺ cells、CFU-GEMM、CFU-GM、CFU-E and BFU-E were detected by using liquid and semi-solid culture systems.RESULTS:TGF-β₁ antisense PS-ODNS cooperated with cytokines increased the number of CD₃₄⁺ cells, CFU-GEMM, CFU-GM, CFU-E and BFU-E, which was as 4, 2.6, 2.7, 1.8, 2.1 times as that of the control (the cytokines combination), respectively. TNF-α antisense PS-ODNS cooperated with cytokines respectively increased the number of CD₃₄⁺ cells, CFU-GEMM, CFU-GM, CFU-E and BFU-E by 4, 2.9, 2.6, 1.7, 1.8 times as that of the control. The above two antisense PS-ODNS cooperated with cytokines could respectively increased the number of CD₃₄⁺ cells, CFU-GEMM, CFU-GM, CFU-E and BFU-E by 5.3, 2.1, 2.7, 1.9, 1.8 times as that of the control.CONCLUSION:Inhibition of endogenous TGF-β₁ and TNF-α by antisense PS-ODNS will be one of the effective methods to expand HSPC ex vivo.     

Effect of cilazapril on pulmonary vascular endothelial dysfunction in hypoxic rats

AIM:To study the effect of cilazapril on pulmonary vascular endothelial dysfunction in hypoxic rats. METHODS:The structure and function of endothelium in hypoxic rats were studied by biochemical analysis, radioimmunoassay, transmission electron microscope and correlated with hemodynamic. RESULTS:1) The change and damage of ultrastructure in endothelial cell (EC) were obsevered in hypoxic rats. 2) The contents of plasma nitric oxide (NO) and superoxide dismutase (SOD) activity in blood as well as endothelial nitric oxide synthase (eNOS) activity in the lung tissue were significantly lower in the hypoxic rat than those in contral animals. The concentrations of plasma endothelin-1(ET-1) and angiotensin converting enzyme(ACE) as well as malondialdehyde(MDA) were significantly higher in the hypoxic rat than these in contral animals. The relaxing and contracting factors had a significant positive/negative correlation with mean pulmonary artery pressure (mPAP). 3) Cilazapril significantly decreased the level of ET-1 and ACE and significantly increased the level of NO and activity of eNOS and SOD. At the same time, cilazapril extenuated hypoxia-induced injuries of EC. CONCLUSION:The results indicate that damaging structure and dysfunction of EC existes in hypoxic rats. The cilazapril effectively preventes and treates the chronic hypoxic PH by relieving the injury and improving secretion in EC.     

菟丝子致死薇甘菊

薇甘菊Mikania micrantha是入侵的害草，已经由珠江三角洲扩散到粤东、粤西沿海地区，以及部分山区。在薇甘菊的天敌调查中看到菟丝子寄生于薇甘菊，严重时可以把薇甘菊致死。我们在本所的实验园中栽种薇甘菊，并让其被菟丝子Cusuta chinensis Lam.寄生，结果证实菟丝子在两个月左右，完全可以抑制薇甘菊的生长，最终把薇甘菊致死。这可能是以草治草的一种新途径，但是菟丝子也是我国农作物的一种重要昌，如何利用菟丝子控制薇甘菊的为害，而又不使菟丝子对薇甘菊的伴生植物和鞭它农作物造成为害，是下一步要解决的问题，菟丝子致死薇甘菊的机理更需要深入研究。     

组氨醇脱氢酶抑制剂的研究动态

组氨醇脱氢酶(HDH)是组氨酸生物合成过程中最后两步反应的催化酶。对组氨醇脱氢酶的结构、催化机理、组氨醇脱氢酶抑制剂等方面的研究动态进行了较为详细的综述。     

苏云金芽孢杆菌cry基因在大肠杆菌中表达产物和生物活性

研究了几种Bt cry基因于大肠杆菌(Escherichia coli)中表达产物在pH 10.0的50mmol/L碳酸钠和20mmol/L乙醇胺溶解液中的溶解性 ,发现同样的Cry蛋白在碳酸钠中的溶解度大于乙醇胺。通过胰蛋白酶消化 ,明确Cry1Ca7、Cry1Ia8酶解产物为 38kD多肽 ;Cry1Ie1、Cry1Cb2、Cry2Ab4酶解产物为 41kD多肽 ;Cry1Ac酶解产物为60kD多肽。采用FPLC层析方法对 6种原毒素及其酶解后得到的毒素多肽进行了分离纯化 ,比较了原毒素和毒素的杀虫活性的差异。其结果表明 ,Cry1Ac的原毒素和毒素对棉铃虫初孵幼虫的校正死亡率均为 100% ,Cry2Ab4的原毒素的毒力高于其酶解毒素。     

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.     

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.     

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。