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

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.     

笋用竹害虫天敌资源研究

本文报道了浙江省笋用竹害虫天敌资源调查的结果，共查明天敌资源44种，经鉴定属于螳螂目1科2种，半翅目1科2种，蛇蛉目1科1种，脉翅目1科2种，鞘翅目2科9种，双翅目2科2种，膜翅目9科22种；蛛形纲蜱螨目1科1种，蜘蛛目3科3种，指出了天敌的寄主和分布。     

影响微孢子虫对棉铃虫幼虫致病力的因子

1997年田间调查时，发现一种寄生于棉铃虫的微孢子虫，对棉铃虫具有很强的致病力。为明确环境因子对该微孢子虫及其致病力的影响，测定了温度与紫外线对该微孢子虫及其杀虫效果的影响。结果表明，该微孢子虫孢子耐受温度范围较宽，最高温度上限为55℃。温度对其致病力有显著影响，在一定温度范围内，随着生境温度的升高，微孢子虫对棉铃虫幼虫致病力增强。该微孢子虫对紫外线较敏感，在紫外线照射下易失活而丧失致病力。     

淡紫拟青霉菌料防治大豆胞囊线虫的后效研究

淡紫拟青霉不同菌株的培养菌料，施２２５ｋｇ／ｈｍ２，对大豆胞囊线虫不但当年有４５％～６９％的较好防效，而且第２年和第３年仍然有２２．５％～３７．７％和７．３％～２２．９％的后效，即空胞囊和被寄生的胞囊数量增加。     

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％。     

长寿花胚性愈伤组织的诱导及胚状体再生

 研究了长寿花胚性愈伤组织的筛选，胚状体的诱导发生、发育过程及植株再生。经过对外观及细胞学观察，看出外观质地疏松、颗粒状的淡黄色愈伤组织细胞圆形且形状规则，是胚性愈伤组织。诱导胚性愈伤组织的最适培养基为：MS+2，4-D 2 mg·L^-1 +BA 0.2 mg·L^-1；胚状体的诱导培养基为MS+BA 2 mg·L^-1 +NAA 0.2 mg·L^-1 +活性炭4 g·L^-1+蔗糖30 g·L^-1，胚状体诱导率可达185个胚状体；胚状体的再生培养基为MS+蔗糖30 g·L^-1。利用石蜡切片对胚状体的发生过程进行了观察。     

起始肥料对一品红生长的影响

 研究了生长介质中起始肥料对一品红生长的影响和一品红不同部位矿质养分含量。结果表明： 速效肥比缓效肥更有利于一品红生长并提高品质，速效肥中供试的两种NO₃^--N与NH₄⁺-N比例对一品红生长影响不大。绿色叶片矿质养分含量，除Fe和Mn较高外，与国外推荐的养分诊断指标相一致，说明目前国外推荐的一品红养分诊断部位适合于绿色叶片。     

60%敌草隆·环嗪酮可湿性粉剂的高效液相色谱分析方法

反相高效液相色谱法测定60%敌草隆·环嗪酮可湿性粉剂有效成分含量,流动相为甲醇和水(57∶43),波长为254nm,环嗪酮和敌草隆的标准偏差分别为0.07和0.052;变异系数分别为0.53%和0.11%;平均回收率分别为98.9%和99.2%。线性相关系数分别为0.9999和0.9995。     

The ex vivo expansion characteristic of endothelial progenitor cells

AIM: To explore the ex vivo expansion characteristics of the endothelial progenitor cells (EPCs). METHODS: CD34⁺ cells were selected from umbilical cord blood mononuclear cells (MNC) by MiniMACS system, expanded at the same conditions as that for total MNC, coincubation of CD34⁺ and CD34^- from the same donation for EPCs. In addition, we tested the effect of vessel endothelial growth factor (VEGF) and passage on cell differentiation, expansion kinetics and apoptosis. EPCs were determined and quantified by immunocytochemistry and flow cytometry. RESULTS: Coculture of CD34⁺ and CD34^-,total MNC led to a significant increase in the expansion of CD34⁺ cells compared with CD34 enrichment (P＜0.05). There was a trend toward decreased apoptosis in cultures when early passage was performed once the linear cord like structures appeared. There was no significant effect on apoptosis between with VEGF and without VEGF group (P＞0.05). These differentiated EPCs were stained positive for CD34⁺, von Willebrand factor (vWF), KDR, CD31 and incorporate acetylated low-density lipoprotein (LDL). CD34⁺ and AC133⁺cells accounted for 68.2%±6.3% (n=6) and 57.2%±9.8% (n=6) of attaching (AT) cells at day 7 of culture, respectively. CONCLUSIONS: Coculture of CD34⁺ and CD34^- or culture of MNC enhances ex vivo expansion of EPCs. Early passage decreases apoptosis rate, VEGF has no significant effect on ex vivo expansion of EPCs.     

鸭瘟病毒强毒株在急性人工感染成年鸭病例体内分布规律

5 6只 3月龄四川麻鸭经皮下接种鸭瘟病毒 (DPV)强毒 SC1株 ,成功建立了 DPV感染的急性病理模型 ,并应用PCR方法检测了不同时间 DPV在感染鸭体各组织器官的分布情况。结果表明 ,接种 2 h后 ,即能够从脑、肝、脾、法氏囊、胸腺中检出 DPV DNA;12 h,可从心脏、肝脏、脾脏、肺脏、肾脏、十二指肠、直肠、法氏囊、胸腺、胰腺、脑、胸肌、食管、腺胃、血液、舌、口腔分泌物、皮肤、骨髓和粪便等检测到 DPV的 DNA。检出时间最早和检出率最高的组织器官为肝脏和脑组织。本试验为阐明 DPV的致病机理和应用 PCR方法检测感染鸭体组织中的 DPV提供了重要的实验数据。