弓形虫微线体蛋白MIC3基因的克隆及原核表达

根据编码MIC3的已知基因序列设计并合成一对引物,应用PCR技术从弓形虫RH株的基因组DNA中扩增编码MIC3的全长基因,克隆入pGEX-KG表达载体,转化入E.coli DH5α感受态细胞,经含氨苄青霉素琼脂平板筛选,小量抽提质粒进行酶切、PCR及DNA测序鉴定.然后阳性重组质粒转化入E.coli BL21-CodonPlus,IPTG诱导,表达产物经SDS-PAGE和Western-blot分析鉴定.结果显示,扩增的MIC3基因与GenBank中相应基因序列（AJ132530）的同源性达99.6 %,表达的MIC3融合蛋白表观分子量约为66 ku,且可被兔抗弓形虫免疫血清识别.说明所获得的表达蛋白质具有一定的反应原性,为下一步利用重组蛋白建立弓形虫的诊断方法和研制弓形虫的亚单位疫苗奠定了基础.     

黄连解毒散超微细粉中黄芩苷在家兔体内的药代动力学研究

采用高效液相色谱法测定灌胃给予黄连解毒散超微细粉和细粉的家兔血浆中黄芩苷的浓度,血药浓度-时间数据经药代动力学分析软件(Pharmaceutical Kinetics Software,PKS)处理,比较黄连解毒散超微细粉和普通细粉中黄芩苷在家兔体内的药代动力学参数。结果如下:黄连解毒散超微细粉组、普通细粉组黄芩苷的药代动力学最佳模型均为二室开放模型。黄连解毒散超微细粉主要药动学参数:Ka=0.497 h-1,t1/2α=2.556 h,t1/2β=8.46 h,AUC0→∞=3.401μg.h/mL,Vd=6.752 L/kg,Tpeak=3.005 h,Cmax=0.311μg/mL;黄连解毒散普通细粉主要药动学参数:Ka=0.419 h-1,t1/2α=2.74 h,t1/2β=5.83 h,AUC0→∞=2.611μg.h/mL,Vd=12.890 L/kg,Tpeak=3.645 h,Cmax=0.215μg/mL。与细粉比较,黄连解毒散超微细粉中黄芩苷的相对生物利用度提高了30.26%。上述结果表明,黄连解毒散经超微粉碎后吸收相增大,可显著提高其有效成分黄芩苷的生物利用度。     

甘肃省年有效降水次数的异常特征分析

利用1958-2001年甘肃省45个气象站的逐日降水资料,统计出年有效降水次数,通过经验正交展开(EOF)和旋转经验正交展开(REOF),研究其异常的空间结构及时间演变规律,同时利用墨西哥帽子波变换技术,对甘肃省年有效降水次数进行多时间尺度分析.结果表明:大范围区域一致性异常是甘肃省年有效降水次数的首要特征;甘肃省年有效降水次数异常区域大致分为4个关键区;从总体上看(较大时间尺度),甘肃省年有效降水次数是增加的,而从较小时间尺度看,则表现为增中有减,减中有增的复杂变化.     

Research Progress of Epigenetics Modification in the Process of Skeletal Muscle Cell Proliferation and Differentiation

Skeletal muscle is the most abundant tissue and the main component of muscle in animals.The process of skeletal muscle cell's proliferation and differentiation is the basis of muscle development and it's directly affects the meat production performance of domestic animals.It has been found that epigenetic modification plays an important role in regulating the proliferation and differentiation of skeletal muscle cells.In this study,the effects of epigenetics on skeletal muscle in terms of the effects of DNA methylation on the proliferation and differentiation of skeletal muscle cells,the selection and expression of factors regulated by histone acetylation,the regulation of non-coding RNA,and the effects of chromosome remodeling.Research progress in the process of muscle cell proliferation and differentiation,briefly describes the effects of different modification methods and factors on the two processes of skeletal muscle proliferation and differentiation.At the same time,the methods and means used by predecessor in the study of skeletal muscle proliferation and differentiation were reviewed,and then the role of apparent regulatory factors in skeletal muscle growth was analyzed.The purpose was to further explain the important role of epigenetic modification in the proliferation and differentiation of skeletal muscle,enhanced the understanding of the regulation of skeletal muscle proliferation and differentiation,provided a reference path for integration with animal production,and also provided skeletal muscle.Molecular regulation of such as growth and development provided more reference materials.     

规模化野猪养殖场仔猪腹泻病原菌的分离鉴定和致病性分析

从重庆某一规模化野猪养殖场腹泻仔猪的粪便棉拭子中分离到5株细菌,经形态学观察、染色特性、培养特性、生化特性分析鉴定为大肠杆菌,分别命名为CQ0401、CQ0402、CQ0403、CQ0404和CQ0405。并进行了溶血性、对Vero细胞的细胞毒性、小鼠致病性、抗药性等试验,试验结果表明:这5株大肠杆菌不产溶血素;腹腔接种小白鼠具有高致病性;抗菌谱基本一致:对菌必治(头孢三嗪)、复达欣(头孢他啶)、先锋必(头孢哌酮)敏感,尤其对菌必治敏感性最高,而对其他的药物敏感性较低或不敏感;CQ0401、CQ0403、CQ0404、CQ0405对Vero细胞均无细胞毒性,而CQ0402对Vero细胞具有较强的细胞毒性。     

种子处理对籽瓜细菌性果斑病防治效果的研究

籽瓜细菌性果斑病(Acidovorax avenae subsp.citrulli)是近年来在新疆出现的一种检疫性病害,种子带菌是该病传播的重要方式之一.通过对市售的带有细菌性果斑病的籽瓜种子进行不同温度及不同药剂的处理,经温室种植测定,计算出各种处理方法对籽瓜果斑病的防治效果.结果显示50℃湿热处理30min;60℃干热处理4h,以及2%的HCl处理20min对防治籽瓜细菌性果斑病种子带菌具有良好的效果,各生产单位可以根据实际情况选择合适的方法,以减轻细菌性果斑病的危害和蔓延.     

特异抗体致敏的SPA茵体浓缩新城疫病毒方法在临床诊断中的应用

为提高新城疫临床诊断的准确率,建立了新城疫病毒浓缩方法,将粪便及脑、肺等器官组织病料中的新城疫病毒经特异性抗体致敏的金黄色葡萄球菌(SPA菌)吸附浓缩后,用RT-PCR进行检测.结果表明,所建立的新城疫病毒浓缩方法应用于临床诊断后,可显著提高检测敏感性,从而提高临床诊断的准确率.     

Relationship between expression of MKK-4 and MMP-9 genes in primary hepatic carcinoma with or without invasion and metastasis

AIM：To investigate the expression of mitogen activated protein kinase kinase 4 (MKK-4) and MMP-9 mRNA in primary hepatic carcinoma (PHC), and to analyze its relationship with invasion and metastasis. METHODS：The expression of MKK-4 and MMP-9 mRNA in 34 cases of hepatic carcinoma tissues and adjacent tissues,and in 12 cases of normal liver tissues were detected with RT-PCR. RESULTS：(1) The expression level of MMP-9 mRNA was higher in metastatic cancer tissues than that in other tissuses (P<0.01). (2) There was significant statistical difference among the expression level of MKK-4 mRNA, but the level in metastatic cancer was low (P<0.01). (3) There was no statistical difference among the expression level of MKK-4 or MMP-9 mRNA among the adjacent tissues and normal tissues (P>0.05). (4) MMP-9 mRNA had a tendency to rise as PHC became invasive and metastatic.The expression level of MKK-4 had a tendency to decline in PHC became invasive and metastatic. (5) The expression level of MMP-9 or MKK-4 mRNA had no correlations with tumor volume,or cell differentiation (P>0.05). (6) There were correlations between expressions of MKK-4 and MMP-9 mRNA in PHC (Pearson Correlation, r=-0.925, P<0.01). CONCLUSION：There are high MMP-9 mRNA expression and low MKK-4 mRNA expression in PHC.The expression level of MKK-4 or MMP-9 mRNA is correlated with tumor metastasis.     

Expression of nerve growth inhibiting factor Nogo-A mRNA and protein in the brain ischemic infarction rats

AIM: To investigate the changes of expression of Nogo-A at different time points in brain ischemic infarct rats. METHODS: The model of 80 cases of middle cerebral artery occlusion (MCAO) in rats was established. The expression of Nogo-A mRNA and protein were determined by Western blotting and hybridization, and the relationship between functional scoring and Nogo-A was also evaluated. RESULTS: In the brain of MCAO rats, Nogo-A mRNA expression was decreased on day 3 and increased significantly on day 7. The highest level was observed at the 21th d, keeping the same level at the 28th d. Nogo-A protein expression showed the same results. These results were correlated with the brain function scoring. CONCLUSION: Expression of Nogo-A does not increase in the early stage, but increases significantly in the late stage of MCAO, suggesting that Nogo-A expression may play an important role in the nerve regeneration of brain ischemic injury.