Detection,cloning and expression of bone morphogenetic protein-1 from human osteosarcoma cell lines     

CHEN Dong-ying  ZHU Quan-sheng  LIU Chao  QIU Ju-shi 《园艺学报》2003,19(6):751-756

AIM:To manufacture recombinant protein of the highly conserved domain in human bone morphogenetic protein-1(BMP-1) using gene engineering methods as antigen for making wide spectrum antibody to BMP-1.METHODS:We analyzed the gene sequences and protein structures of BMP-1 and its related proteins, and chose a highly conserved fragment as target gene. Total RNA was prepared from human osteosarcoma cell line Saos-2, then the target gene was amplified with RT-PCR. The PCR product was cloned into prokaryotic expression vector pMAL c2 to get recombinant vector BMP-1(322-588aa)-pMAL c2. After transforming the recombinant plasmid into DH5-alpha and screening, several prositive clones were got for sequencing. Finally the transformed cells was induced with IPTG to get fusion protein.RESULTS:The BMP-1 gene fragment was successfully cloned into vector pMAL c2, and was able to express efficiently with IPTG inducement. The amount of expressed fusion protein is about 66%-72% in total volume of bacterial proteins.CONCLUSIONS:The recombinant protein contains several key domains(2 CUB domains and 1 EGF domain), which are shared by BMP-1 and its related proteins. Specific wide spectrum antibody to human BMP-1 and its related proteins may be generated with this recombinant protein antigen.  相似文献   

Effect of antifibrotic herb serum on rat hepatic stellate cell proliferation and collagen synthesis     

LING Yun-biao  XU Rui-yun  QIU Wan-shou  CHU Zhong-hua  YANG Hong-zhi  TAN Zhi  OU Qing-jia 《园艺学报》2003,19(12):1649-1652

AIM: To investigate the effect of Chinese herbs, Ganxianfang(GXF), on rat hepatic stellate cells (HSC) proliferation and collagen synthesis. METHODS: Two types of herb serum, portal venous serum and circumferential venous serum, were prepared from rats infused intragastrically with 16, 8, 4 times adult dose of GXF decoction. HSC isolated from rat liver were processed with the above sera in vitro. Then we mensurated the radioactivity of HSC admixed with [[³H]H]proline and [[³H]H]thymine to judge the effect on proliferation and collagen synthesis of HSC. RESULTS: Both two types of serum collected 0.5, 1, 2 h after intragastrical infusion inhibited HSC proliferation (P＜0.05), and the serum collected 1 h after intragastrical infusion had the strongest effect (P＜0.05). Portal serum decreasea collagen synthesis (P＜0.05), but circumferential serum had no effect (P＞0.05). CONCLUSION: Inhibition of HSC proliferation and decrease of collagen synthesis may contribute to the GXF antifibrotic action.  相似文献   

Inhibitory effect of berberine on the activation and proliferation of T lymphocytes     

HE Xian-hui  ZENG Yao-ying  XU Li-hui  QIU Hai-xia  CAI Xiao-chang 《园艺学报》2002,18(10):1183-1186

AIM: To investigate the effect of berberine (Ber) on the activation and proliferation of T lymphocytes and its mechanism of action. METHODS: Whole peripheral blood from normal subjects was stimulated with phytohemagglutinin (PHA) or phorbol ester (PDB) plus ionomycin (Ion) and the expression levels of CD69 and CD25 were evaluated with flow cytometry after the staining with appropriate fluorescent monoclonal antibody. The distribution of cell cycles was analyzed by propidium iodide staining and dead cells by 7-aminoactinomycin live staining. RESULTS: 100 μmol/L and 50 μmol/L of Ber had significant inhibition of the expression of CD69 on T cells stimulated with PDB plus Ion or PHA, while effect of 25 μmol/L Ber was not significant. And as time of action extended, the extent of inhibition decreased. For the expression of CD25, Ber at the concentrations as above all exerted significant inhibitory effect in a dose-dependent manner. Moreover, Ber could block lymphocytes cell cycle progression from G₀/G₁ phase to S and G₂/M phase without phase specificity. Besides, live staining analysis revealed that Ber did not have significant cytotoxicity on lymphocytes. CONCLUSIONS: Ber significantly inhibits the expression of early and mid activation antigens of T cells and also blocks the progression of lymphocytes cell cycles. These results suggest that Ber exerts immunosuppression effect through inhibiting the activation and proliferation of T cells.  相似文献   

昆虫病原线虫rDNA多态性分析   总被引：1，自引：0，他引：1  

丘雪红韩日畴  许再福 《昆虫天敌》2003,25(3):130-144

本文对国内外昆虫病原线虫斯氏属和异小杆属的47个品系进行rDNA—ITS PCR—RFLP分析，研究其DNA多态性，并构建了分子系统发育树状图。各品系的ITS区无明显的长度差异，PCR—RFLP分析将47个品系分为斯氏属和异小杆属两大类，两属线虫又分别分为11组和4组。所得结果丰富了ITS PCR—RFLP图谱库，为弄清我国的昆虫病原线虫与国外种类的分子系统发育关系及未定名线虫的鉴定提供重要依据，同时为筛选适合的线虫种类防治害虫奠定基础。  相似文献   

肉仔鸡生长过程中行为与产热的变化     

任建军  高文婷  邱泽文  王靖宇 《畜牧与兽医》2004,36(1):12-14

研究肉仔鸡成长过程中行为变化对产热量的影响 ,将 2日龄肉仔鸡 36只分为自由摄食、 50 %摄食和绝食 3组进行观测。立位行为集中在明期 ,特别是摄食 30min前后 ,暗期几乎没有出现立体行为。 2日龄时 ,每天约有 70 0min是立位的 ,其后 ,随着日龄的增加而减少。在自由采食区 ,公雏比母雏摄食时间长。 50 %摄食区较自由采食区的摄食时间少。在绝食区 ,随着日龄的增加 ,公母的立位时间呈现减少倾向。 2 1日龄之前随着日龄的增加其摄食时间也增加 ,其后呈现相反倾向。产热 (HP ,kJ/kg0 75)与摄食时间有相同倾向。在 2 1日龄前公雏的HP比母雏低 ,2 1日龄后母雏比公雏低。 1天中各个区的HP活动规律是摄食后 1h内最高 ,暗期降低 ;限制摄食量后 ,HP降低 ,明期和暗期的HP差也随之减小  相似文献   

徐州地区烟粉虱发生规律及治理措施初探   总被引：4，自引：0，他引：4  

戴率善  刘宗泉  杜予州  周福才  张贵永  高素兰  仇保华  李建树  李艳红 《中国植保导刊》2004,24(3):17-20

通过近2a观察表明,徐州地区烟粉虱在田间消长大致分4个阶段,成虫有4个主要迁移期;其喜干热、耐高湿的生活习性、冬季保护地栽培面积的扩大、外来虫源的传入及防治不力是烟粉虱种群数量累积并引起暴发的主要原因;主要寄主有9科20余种栽培作物;寄主植株上虫量垂直分布:棉花上部>中部>下部,温室蔬菜一般中、下部>上部。可采取严格检疫、清洁田园、轮作换茬、黄板诱杀、覆盖防虫网、保护和利用天敌、药剂防治等措施进行治理。  相似文献   

内生拮抗枯草芽孢杆菌BS-2菌株的发酵条件   总被引：26，自引：1，他引：26  

何红  沈兆昌  邱思鑫  蔡学清  关雄  胡方平 《中国生物防治》2004,20(1):38-41

内生拮抗细菌BS 2菌株在以黄豆粉为原料的 3号培养基中生长速度快 ,发酵滤液对辣椒炭疽病菌的抑制作用强 ,菌液对辣椒果炭疽病的防效最好。培养基初始 pH值、培养时间、温度、通气量等对菌株生长及其抗菌物质的分泌有明显影响。结果表明 ,以黄豆粉培养基、初始 pH值 6 .7(灭菌后 )、2 8℃、培养 4 8h、并尽量增大培养通气量 ,为菌株的最佳发酵条件。  相似文献   

Effects of component of some Chinese herbs on proliferation of human umbilical vein endothelial cells in vitro     

HE Xiu-juan  LI Ping  QIU Quan-ying  SHENG Xun  WANG Fang 《园艺学报》2004,20(5):832-835

AIM: To observe the effects of some component of Chinese herbs for external use on proliferation of human umbilical vein endothelial cells (HUVEC) and investigate the mechanism of promoting tissue repair. METHODS: The method of MTT was used to examine the effects of Rg1, Rh1, perlolyrine, cinnamyl aldehyde, muscone, astragaluspolysaccharin (APS), velver antler polypeptide (VAP) and soluble extract of boswellia carterii birdw (BCB) on proliferation of HUVEC. RESULTS: APS did not promote proliferation of HUVEC at 9.75 mg/L-2.5 g/L; Rh1 promoted proliferation of HUVEC at 1.94 mg/L-0.5 g/L (P<0.05 or P＜0.01), and Rg1 inhibited proliferation of HUVEC at 31 mg/L (P<0.05); VAP promoted proliferation of HUVEC at 1 mg/L-0.5 g/L with optimal dose of 10 mg/L (P<0.01), Cinnamyl aldehyde promoted proliferation of HUVEC at 2 g/L(P＜0. 05); Muscone and soluble extract of BCB inhibited proliferation of HUVEC at 1 g/L, 0.5-2.5 kg/L(P＜0. 01), respectively; Perlolyrine inhibited proliferation of HUVEC at 0.125 g/L-0.5 g/L(P＜0. 01). CONCLUSION: The external herbs for supplementing Qi and warming Yang can promote HUVEC proliferation and improve angiogenesis during tissue repair. The external herbs for promoting blood circulation and accelerating capillary movement may have influence upon other stages of tissue repair.  相似文献   

The molecular mechanism of inhibition of murine Lewis lung carcinoma metastasis by weimaining in vivo     

LOU Jin-li  LIN Hong-sheng  QIU Quan-ying  PEI Ying-xia  QI Xin  HE Xiu-juan 《园艺学报》2004,20(4):627-630

AIM: To investigate the anti-metastasis effect of weimaining, extracted from fragopyrum cymosum meissn, a Chinese medicine, on murine Lewis lung carcinoma (3LL). METHODS: The anti-metastasis effect of weimaining in vivo was detected in the grafting lung metastasis model of murine Lewis lung carcinoma. The effects of the drug on the expression of CD34 and E-cadherin were investigated by immunohistochemical staining and RT-PCR. RESULTS: Weimaining effectively inhibited the lung metastasis of 3LL at a concentration of 250 mg·kg^-1·d^-1, significantly suppressed the expression of CD34 and increased the expression levels of E-cadherin protein and mRNA in 3LL cells. CONCLUSIONS: Weimaining inhibits the metastasis of murine Lewis lung carcinoma (3LL) in vivo via increasing the expression of E-cadherin and decreasing microvessel density of tumor tissue.  相似文献   

抗鸡传染性喉气管炎重组鸡痘病毒基因工程疫苗同居感染试验   总被引：3，自引：0，他引：3  

王云峰  智海东  王玫  孙永科  仇华吉  周艳君  田志军  张绍杰  童光志 《中国预防兽医学报》2004,26(1):51-54

疫苗的接触传播是疫苗免疫接种需要考虑的重要因素,为了检测重组鸡痘病毒载体疫苗水平传播的能力,对隔离条件下饲养的SPF鸡用重组鸡痘病毒基因工程疫苗接种,同时设立非免疫对照鸡,饲养期间特意延长清粪时间以增加感染的机会,1个月之后攻击传染性喉气管炎WG株强毒和鸡痘102株强毒,疫苗免疫鸡全部获得保护,而非免疫鸡则全部发病.在试验动物饲养场的自然条件下,将免疫鸡和试验对照两组鸡饲养在同一个鸡舍内,让疫苗毒的传播更接近自然条件.在每个月的攻毒试验中,对照鸡都没有获得对鸡痘和传染性喉气管炎强毒的保护.在疫苗免疫期间进行连续5个月的跟踪检测,同居未免疫鸡没有检测到抗传染性喉气管炎病毒gB抗体.这些实验结果表明抗鸡传染性喉气管炎重组鸡痘病毒基因工程疫苗不能通过接触传播.  相似文献