植物提取物辐照杀菌工艺研究

检测了几种植物提取物中污染微生物的种类和数量 ,研究了当归提取物中污染微生物的D10 值 ,得出植物提取物辐照杀菌工艺的最低有效剂量为 4 0kGy,最高耐受剂量为 8 0kGy。     

Construction and Identification of Yeast Surface Display Libraries of Ovis aries DRA Gene Exon 2

The specific primers were designed according to Ovis aries DRA gene sequence deposited in GenBank and the multiple cloning site of the plasmid pYD1,which was a vector used for protein surface display on Saccharomyces cerevisiae.The gene encoding DRA was amplified by PCR using the genomic RNA of Ovis aries.The 762 bp fragment was cloned and released in GenBank and registration number was KR422362.The PCR product was inserted into the yeast surface display plasmid vector pYD1 by double enzyme digestion.It was indicated that DRA gene was successfully integrated into the genome.Dot mutation was made at both ends of exon 2 in DRA gene for making restriction enzyme cutting site and design the exon 2 specific primers according to mutated Ovis aries DRA gene sequence.Sequenced exon 2 amplification products based on DNA pooling of sheep large sample template was analyzed the polymorphic loci.The polymorphic exon 2 246 bp fragment was obtained by double enzyme digestion and connected to surface display restructuring mutation carriers pYD1-DRA by the same double enzyme digestion,and then we successfully constructed yeast surface display libraries.We transformed it into Saccharomyces cerevisiae EBY100 cell.Yeast monoclone was identified by PCR amplification and sequencing,and we confirmed that DRA gene had been integrated into Saccharomyces cerevisiae genome.After galactose induced,it was detected that DRA gene library had been successfully demonstrated on the yeast cell surface under the fluorescence microscope by immunofluorescence method.     

Hmgn3和Hoxa10对小鼠子宫基质细胞蜕膜化过程中Foxo1表达的调控

利用小鼠子宫基质细胞体外诱导蜕膜化模型转染Hmgn3或Hoxa10过表达载体及siRNA片段,通过荧光定量PCR方法检测Hmgn3和Hoxa10对Foxo1表达的调控。结果显示:转染Hmgn3或Hoxa10过表达载体可促进Foxo1在子宫基质细胞蜕膜化过程中的表达,而转染Hmgn3或Hoxa10siRNA则可抑制Foxo1的表达。在小鼠子宫基质细胞中转染Hmgn3或Hoxa10siRNA后再添加孕酮,Foxo1的表达显著下降。同样,干扰Hmgn3或Hoxa10也可减低cAMP对Foxo1表达的调控。结果表明:Hmgn3和Hoxa10可通过Foxo1来影响小鼠子宫基质细胞的蜕膜化,孕酮和cAMP可通过Hmgn3和Hoxa10来调控Foxo1在子宫基质细胞中的表达。     

四种牛分枝杆菌特异性蛋白融合表达及在牛结核病诊断中的临床应用

以原核表达的MPB70-MPB83-CFP10-ESAT-6融合蛋白作为诊断抗原,建立牛结核病抗体检测间接ELISA(iELISA)。该iELISA与导致常见牛病的非相关抗原无交叉反应。用iELISA检测90份健康牛血清,确定样本阴阳性临界值(S/P)为0.17。与商品化胶体金试剂条(ICG)平行检测150份临床奶牛血清样本,结果与ICG的符合率为93.33%(140/150)。以结核菌素皮内试验(TST)为参考方法,检测了华中地区四个奶牛场的560头中国荷斯坦奶牛和90份进口奶牛结核阴性血清,结果iELISA与TST的总符合率为87.32%(489/560)。对其中22头奶牛进行鼻拭子分菌检验,4头分菌阳性奶牛的血清抗体检测也为阳性,iELISA与细菌培养的总符合率为77.27%(17/22)。以TST为参考方法,iELISA的敏感性和特异性分别为72.37%和89.67%。     

蜂王浆中10-HDA的研究进展

对蜂王浆中10-HDA的来源、结构、性质及功能等方面的研究进展进行了介绍，并对近年来不断改进和完善的几种测定方法，如分光光度法、色谱法、酶联免疫法、气相色谱-质谱法等进行了简要分析。     

牛分支杆菌CFP-10的原核表达及其在牛结核病诊断中的应用

低分子质量的蛋白抗原CFP-10是一种重要的牛分支杆菌早期分泌蛋白.为了检测该蛋白和其他几种抗原在牛结核病诊断中的临床应用,对CFP-10的基因进行克隆,鉴定,并在原核系统中表达.PCR法扩增cfp-10基因片段,连接到pET-22b( )原核表达载体中,再转入表达宿主E.coli BL21(DE3)PlysS菌株内,用IPTG诱导,进行蛋白表达、纯化.分别以CFP-10、ESAT-6、MPT83、MPT70、牛PPD、CFP-10与ESAT-6混合蛋白,MPT83与MPT70混合蛋白为抗原,用间接ELISA法诊断牛结核病.结果表明,以CFP-10与ESAT-6混合蛋白作为抗原检测牛结核病的特异性和敏感性分别达到了100%和63.6%,均超过了其他单抗原或抗原组合,为以筛选合适抗原为基础的血清学诊断技术提供了有力的支持并创造了良好的应用前景.     

Recombinant hFGF-10 adenovirus promotes proliferation of kerotinocytes

AIM:To construct the recombinant adenoviral vector containing human fibroblast growth factor 10 (hFGF-10) gene, and to study the effect of the recombinant adenovirus on the proliferation of kerotinocytes. METHODS:HFGF-10 gene was amplified by PCR and ligated with shuttle vector pAdTrack-CMV to get the recombinant plasmid pAdTrack-CMV-hFGF-10, which was linearized with PmeI and transferred into Escherichia coli BJ5183 containing the adenoviral bone plasmid pAdEasy-1 for homologous recombination to obtain the recombinant adenoviral plasmid pAdEasy-hFGF-10. The recombinant adenoviral plasmid was then transfected into HEK-293 cell line to package and amplify the recombinant adenovirus. The expression of hFGF-10 in HaCat cells infected with the recombinant adenovirus was detected by Western blotting. The influence of the recombinant adenovirus on the proliferation of kerotinocytes was checked by MTT. RESULTS:The recombinant adenovirus containing hFGF-10 gene was successfully constructed, which effectively infected HaCat cells. The result of Western blotting showed that a protein in culture media of the infected HaCat cells reacted with hFGF-10 antibody. The recombinant adenovirus stimulated the proliferation of kerotinocytes. CONCLUSION:HaCat cells infected with the recombinant adenovirus expresses and secrets hFGF-10 protein, which promotes the proliferation of HaCat cells.     

Effects of ganmao shuangjie heji on the inflammatory damage in the lungs of mice infected by IV FM1

AIM: To establish the influenza infected mouse model and study the anti-inflammatory effect of ganmao shuangjie heji. METHODS: IV FM1 infected mice were used as the animal model. The changes of pathology and the cytokine TNF-α, IFN-γ and IL-10 in the lung were observed by HE staining and ELISA (double antibody sandwich enzyme linked immunosorbent assay) after ganmao shuangjie heji treatment. RESULTS: After infected by influenza virus, severe interstitial pneumonia was induced in the model group. Mild interstitial pneumonia was observed in ganmao shuangjie heji treated group. The protein expressions of cytokine TNF-α, IFN-γ and IL-10 were higher in model group than those in the control group. The protein expressions of TNF-α and IFN-γ in ganmao shuangjie heji treated group decreased and IL-10 expression increased significantly compared with model group. CONCLUSION: Ganmao shuangjie heji decreases the expressions of TNF-α and IFN-γ, and increases the expression of IL-10, thus, alleviates inflammatory injury. The clinical application of this medicine can shorten the course of disease.     

菜用大青豆AC10的选育

ＡＣ１０大豆是以７５－２３－１－５－１５品系经激光红宝石辐照,选衣出７５－５４红品系,以之为父本,以西德青豆ＳｏｇｌｅｅｎＯｇｄｅｎ为线本进行有性杂交,再用红宝石２５００Ｖ辐射选能而成,具早熟、高产、优质等特点,６６７ｍ＾２产量１５４．４７ｋｇ,比对照八个月爆增产３０．６６％。     

中椒10号的选育

中椒１０号系以９００１３－３和７８－１－１两个自交系配制而成的微辣型一代杂种,熟性早,早期产量比对照品种增加２．４％～１２．５７％,结果性好,抗ＴＭＶ,中抗ＣＭＶ。一般每６６７ｍ２产２５００～３５００ｋｇ,平均单果重３０．９～４２．５ｇ,果实长羊角形,果面光滑,颜色绿,肉质脆嫩,味微辣,品质佳,果实商品率高。已在北京、天津、河北、东北、安徽、四川及广东、广西、海南等南菜北运基地推广,面积达４８００ｈｍ２。