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   

Changes of K⁺ channels of outer hair cells in guinea pig cochlea with streptomycin ototoxicity     

CUI Gui-ying  TANG Hao  WU Yu-huan 《园艺学报》2002,18(6):619-621

AIM:To study the changes of K⁺ channels of outer hair cells in guinea pig cochlea with streptomycin ototoxicity. METHODS:Auditory brainstem responses (ABR) and whole-cell patch clamp techniques were used.RESULTS:(1) The body weight of guinea pigs with streptomycin ototoxicity decreased significantly; (2) The ABR threshold markedly increased in streptomycin group (Ⅱ,Ⅲ);(3)The number of dissociated outer hair cells of guinea pigs (Ⅱ,Ⅲ) was lower than that of control (Ⅰ); (4) Streptomycin decreased the Ca²⁺-sensitive K⁺ currents and delayed outward K⁺ currents distinctly; (5) There was no significant difference of K⁺ currents between Ⅰ and Ⅱ/Ⅲ. CONCLUSION:These results suggest that the inhibition of K⁺ channels is the basis of streptomycin ototoxicity, but not the direct reason for cell death.  相似文献   

Establishment of chronic obstructive pulmonary disease rat models by passive cigarette smoking and intratracheal instillation of lipopolysaccharide     

LI Hong-mei  CUI De-jian  TONG Xin  PANG Bao-sen  GAO Ya-bing  WANG De-wen 《园艺学报》2002,18(7):808-812

AIM:To establish rat chronic obstructive pulmonary disease(COPD) models by passive cigarette smoking plus intratracheal instillation of lipopolysacchride(LPS) or passive cigarette smoking only, which would be similar to the pathogenesis of human COPD. METHODS:48 Wistar rats were randomly divided into 4 groups.(1) Healthy control I group(n=12), rats were bred 4 weeks;healthy control II group(n=12), rats were bred for 3months. (2) Model group I (n=12), 200μg lipopolysaccharide(LPS) was instilled intratracheally once for every two weeks and the rats were exposured to 5% of cigarette smoke, 0.5 h/d for 4 weeks.(3) Model group II(n=12),rats were exposed to 5% of cigarette smoke, 0.5 h/d for 3 months. The pathologic changes of airways and lung tissues, pulmonary function and blood gas analysis were determined. The airway wall lymphocytes and alveolar macrophages were counted. The cross areas of epithelial layer, smooth muscle layer and lamina propria of bronchi were measured. The hydroxyproline of lung tissue homogenates was determined by biochemistry method.RESULTS:The pathologic changes of airways and lung tissue of two models were similar to but milder than those of COPD patients(biopsy data). The collagen deposition and the cross areas of epithelial layer and smooth muscle layer in airway walls of two model groups were significantly increased than those of control groups(P＜0.01,P＜0.05).FEV_0.3/FVC% of two model groups, PaO₂ and SaO₂ of model I group were significantly decreased, while R_i and R_e in model I group were significantly increased than that of control I group(P＜0.05). The PaCO₂ and the counts of lymphocytes and alveolar macrophages of both model groups were significantly increased than those of the control groups (P＜0.01). Lots of alveolar macrophages had phagocyted smoke granules. The amounts of hydroxyproline of two model groups were significantly increased than those of control group((P＜0.05) and were negatively related to the FEV_0.3/FVC%, respectively (P＜0.01,P＜0.01) and positively related to airway resistance of model I group(P＜0.01). CONCLUSIONS:COPD rat models were successfully established by passive cigarette smoking plus intratracheal instillation of LPS and cigarette smoking only. The pathologic changes were similar but milder than those of COPD patients. The airway obstruction of model I group was more severe than that of model II group, but they have no significant difference.  相似文献   

Effects of platelet-derived growth factor on DNA and collagen protein synthesis in human vascular fibroblasts     

REN Yu-sheng  WU Zong-gui  CUI Fang  JIA Guo-liang  YU Shi-qing  TANG Chao-wu  LI Bo 《园艺学报》2002,18(11):1377-1380

AIM: To investigate the effects of platelet-derived growth factor on DNA and collagen protein synthesis in human vascular fibroblasts. METHODS: In the present experiment, the human vascular fibroblasts were cultured and effects of platelet-derived growth factor-BB on DNA and collagen protein synthesis in human vascular fibroblasts were observed by using [³H]-TdR incorporation and [³H]-proline incorporation in vitro. RESULTS: Platelet-derived growth factor-BB significantly promoted NDA synthesis and collagen protein synthesis of quiescent human vascular fibroblasts, with a maximal response at a concentration of 30μg·L^-1at 24 h and 36 h, respectively. CONCLUSION: Platelet-derived growth factor-BB promotes DNA and collagen protein synthesis in cultured human vascular fibroblasts.  相似文献   

笋用竹害虫天敌资源研究   总被引：1，自引：0，他引：1  

徐华潮骆震旦  王信培张慧 《昆虫天敌》2003,25(3):119-124

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。  相似文献   

施氮量对棉田主要害虫种群动态的影响   总被引：5，自引：0，他引：5  

崔金杰  夏敬源  王春义  马丽华  雒君瑜 《中国植保导刊》2003,23(1):3-5

通过不同施氮肥量对棉田主要害虫种群动态影响的研究结果表明,在低、中、高3种施氮肥水平处理条件下,随着氮肥量的增加,棉田棉铃虫落卵量增加,棉苗蚜、棉叶螨种群数量减少,发生为害减轻,但棉伏蚜种群数量增加。中等施氮肥条件下,第二、三代棉铃虫幼虫数量高于低和高施氮肥处理,对棉铃虫幼虫的生长发育和存活较为有利。  相似文献   

淡紫拟青霉菌料防治大豆胞囊线虫的后效研究   总被引：6，自引：2，他引：6  

王昌家  张新德 《中国生物防治》1997,13(1):26-28

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。  相似文献   

国产克洛素隆尿药排泄数据分析     

胡振英  史彦斌  张新国  罗永江  罗超应  尚若峰  梁纪兰  崔颖  郑继方 《中兽医医药杂志》2004,23(4):9-11

应用Kromasil-C18色谱柱(250 mm×4.6 mm,5 μm),WatersTM480型可调波长紫外检测器,0.01M磷酸钾(pH=7):乙腈(3:1)为流动相,检测波长265 nm,含量测定采用标准曲线法,建立了RP-HPLC法检测绵羊尿中克洛素隆含量的方法.方法有效性评价结果表明,尿药含量在0.01～5.0μg/ml及5.0～30.0μg/ml范围呈良好线形关系(r=0.9993、0.9995),方法平均回收率99.32%,日内、日间变异系数分别为3.91%、6.28%.尿药最低检测限0.005μg/ml.试验绵羊以7 mg/kg单剂量经静脉、肌肉及口服三种途径给药后,尿中药物浓度分析表明,克洛素隆经体内处理后主要经肾脏排泄,给药96 h内,静注经尿排泄原药为81.76%,肌注为64.03%,口服为48.50%.  相似文献   

长寿花胚性愈伤组织的诱导及胚状体再生   总被引：10，自引：0，他引：10  

陈超  王桂兰  田立民  崔瑞生 《园艺学报》2004,31(2):249-252

 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。  相似文献   

起始肥料对一品红生长的影响   总被引：2，自引：1，他引：2  

焦晓燕  孙景桐  关超  任志强  刘惠民 《园艺学报》2004,31(3):389-391

 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。  相似文献