Construction of recombinant retroviral vector of short interfering RNAs specific for macrophage migration inhibitory factor (MIF) and establishment of stable HeLa cell line with a persistent knockdown of MIF     

DAI Bo  XIAO Ding-zhang  YU Xi-yong 《园艺学报》2008,24(11):2195-2199

AIM: To construct recombinant retroviral vector of short interfering RNAs (siRNA) specific for macrophage migration inhibitory factor (MIF) and to establish the stable knockdown of MIF cell line of mammalian cells by transfecting the recombinant retroviral vectors. METHODS: We synthesized oligo-nucleotides for MIF in vitro, and cloned them into retroviral vector pSuper.retro. Subsequently the plasmids were sequenced and digested to identify the construction of the recombinant retroviral vectors. The vectors RNAi were transfected into packing cell line PHOENIX, which was selected by puromycin later. HeLa cell line was infected by the virus supernatant of stable PHOENIX cell lines, and the stable HeLa cell line showed significantly to silence MIF was established by selecting with puromycin. We also compare the characters of HeLa-pSuper-mock to HeLa-pSuper-MIF cells by using migration assay, adhesion assay, soft agar assay and FACS analysis of the cell-cycle progression. RESULTS: The recombinant retroviral vectors were constructed successfully. The HeLa cell line infected by the supernatant containing the retrovirus of package PHOENIX cells was persistent knockdown of MIF confirmed by Western blotting. Knockdown of MIF in HeLa cells inhibited the migration and adhesion, and decreased the clone formation. FACS analysis revealed that knockdown of MIF arrested HeLa cells in G0/G1 phase. CONCLUSION: We establish the stable HeLa cell line with a persistent knockdown of MIF. Our current studies reveal that MIF is necessary for HeLa cell migration and anchorage-independent growth.  相似文献   

Change of β₂-adrenoceptor and its effect on stretch-induced arrhythmias in rat after myocardial infarction     

XIE Rong-sheng  FU Lu  CAO Jun-xian  DAI Ying-nan  ZHU Li-qun  HAN Ying 《园艺学报》2008,24(4):809-811

  相似文献   

第三代果树及开发利用价值     

刘德兵  魏军亚  代正福  王樱莅 《北方园艺》2008,(4):88-90

综述了第三代果树的概念、开发利用价值、国内外研究进展以及开发利用的原则和途径.  相似文献   

几种苏云金芽孢杆菌杀虫晶体蛋白对黑龙江省主要蔬菜害虫小菜蛾活性分析   总被引：1，自引：0，他引：1  

韩岚岚  戴长春  宋福平  张杰  赵奎军 《北方园艺》2008,(8)

利用已克隆的3种Bt cry基因cry1Ca、cry1Ia、cry2Ab和1种野生菌株HD-73(cry1Ac)表达的4种Bt Cry杀虫晶体蛋白,对小菜蛾进行生物活性分析,并将Cry1Ac杀虫晶体蛋白分别与其它3种蛋白按1:1的比例组合,对小菜蛾进行生物活性测定.结果表明:单独使用Cry1Ac的活性最高,LC50为2.39 μg/mL,Cry1Ac与Cry2Ab混合对小菜蛾的具有较高的毒力,LC50为48.91,高于其它组合.  相似文献   

早春大棚厚皮甜瓜品种比较试验   总被引：2，自引：1，他引：1  

GUO Fengling WANG Hongsheng DAI Zhaoyi LI Jinquan 《长江蔬菜》2008,(8)

通过采用湖北省普遍采用的大棚爬地栽培方式,对12个厚皮甜瓜品种进行品种比较试验。结果表明,银蜜子、蜜世界、状元、红妃、金帝等品种在品质、熟性、抗病性及产量等方面的综合表现较好,适合湖北省早春大棚栽培。  相似文献   

连栋拱形塑料温室拱轴线的简易优化法     

曹保山 《长江蔬菜》2008,(10)

在考虑雪栽的情况下,依据结构力学的基本原理,提出了一种不经过有限元的复杂计算、概念明了、方法简易的确定温室拱轴线的新方法.利用该方法对温室的拱轴线进行研究,发现雪载对拱半径的影响很小,拱半径随温室单跨增大而增大,拱半径与矢跨比的关系比较复杂,并为快速确定拱半径参考值提供一张表.  相似文献   

黑木耳原生质体制备、再生及单核体荧光鉴定   总被引：1，自引：0，他引：1  

韩增华  张丕奇  戴肖东  孔祥辉  马庆芳  张介驰 《食用菌学报》2008,15(3)

利用正交试验研究了黑木耳原生质体制备、再生的最佳条件,结果表明,原生质体制备最佳条件是酶解温度31℃、酶浓度1．0％、菌龄5d、酶解时间4h,原生质体再生的最佳条件是酶解温度27℃、酶解时间4h、酶浓度1．5％、菌龄11d。稳渗剂种类对黑木耳原生质体再生率的试验结果表明,采用0．5mol／L蔗糖为稳渗剂,各黑木耳菌株的原生质体再生率最高。HW2号菌株原生质体经核染色其单核化比率最高;在荧光显微镜下观察菌丝核相,单、双核清晰可辨。  相似文献   

全基因组基因表达分析技术及其在果树上的应用   总被引：1，自引：1，他引：0  

张志宏  赵进春  代红艳 《果树学报》2008,25(3):382-388

基因表达分析对于揭示基因功能、了解基因间相互关系、阐明植物生长发育过程中的生理生化调控机制起着至关重要的作用。综述了mRNA差异显示、cDNA-AFLP、基因芯片、EST数据分析、基因表达系列分析(SAGE)等全基因组基因表达分析技术的原理特点及在果树上的应用情况,并介绍了利用全基因组基因表达分析技术开展果树研究的思路。  相似文献   

Change of intestinal flora and its relationship with IL-23/IL-17 axis in patients with ulcerative colitis     

MA Xu-yuan  DAI Zhi-feng  WANG Hui-chao  YANG Jing-nan  TANG Xiang-yang  KANG Yu-hua  DING Chun-sheng  LI Yu-xia  YANG Rui-lin  LIN Xu-hong 《园艺学报》2018,34(5):884-892

AIM:To investigate the change of intestinal flora distribution and its relationship with interleukin-23 (IL-23)/IL-17 axis in ulcerative colitis (UC) patients. METHODS:The fresh fecal samples from 20 patients with active UC and 20 healthy controls were collected. The distribution of the flora was analyzed by direct smear and traditional bacterial culture. The changes of bacteria were detected by real-time PCR. The hemoglobin, albumin, erythrocyte sedimentation, and C-reactive protein levels were tested routinely. Both normal and damaged mucosal tissues of UC patients were examined and obtained by colonoscopy, and further assessed by Mayo scoring, Baron grading and HE staining. The expression of IL-17 and IL-23 was observed by immunohistochemistry and Western blot. RESULTS:(1) The degree of flora imbalance in active UC patients was higher than that in the healthy controls (P<0.05). (2) The results of aerobic culture showed that the number of Escherichia coli in the UC patients was significantly lower than that in the normal controls (P<0.01), while Enterococcus was increased obviously (P<0.01). The results of anaerobic culture revealed that the numbers of Bacteroidetes, Bifidobacterium bifidum and Lactobacilli in the UC patients were significantly decreased (P<0.01). (3) Quantitative analysis of target bacteria showed that the relative quantification of Escherichia coli, Bacteroidetes, Bifidobacterium bifidum and Lactobacilli in the UC patients was significantly lower than that in the normal subjects, and the number of Enterococcus was significantly increased (P<0.01). (4) Compared with control group, no significant change of hemoglobin in the UC patients was ovserved, albumin was significantly decreased (P<0.05), but erythrocyte sedimentation and C-reactive protein levels were elevated obviously (P<0.01). (5) The Mayo score, Baron grade, and histopathological score were all increased (P<0.01). (6) High IL-17 and IL-23 expression levels were detected in the UC patients (P<0.01). (7) Correlation analysis showed that the average absorbance values of IL-17 and IL-23 expression were positively correlated with Baron grade (r=0.717, P=0.02; r=0.849, P=0.016) and pathological score (r=0.660, P=0.03; r=0.675, P=0.032). Meanwhile, the average absorbance value of IL-23 expression was negatively correlated with the number of Escherichia coli (r=-0.699, P=0.025), and positively correlated with Enterococcus (r=0.872, P=0.010). Furthermore, the average absorbance value of IL-17 expression was positively correlated with Enterococcus (r=0.764, P=0.046), and both of them were not correlated with other bacteria. CONCLUSION:Obvious flora imbalance exists in active UC patients, changed intestinal microflora is closely related with the degree of inflammation. IL-23/IL-17 axis, as a key factor in the development of UC, may be related to the changes of intestinal microflora. The interaction between intestinal microflora and IL-23/IL-17 axis plays an important role in the pathogenesis of UC.  相似文献   

几种新型杀菌剂对菜豆炭疽病菌的室内毒力测定     

戴启星祝忠有  张树武徐秉良 《中国蔬菜》2018,1(12):49-53

采用生长速率法,测定了咪鲜胺锰盐、福·福锌、苯醚·溴菌腈、二氢蒽醌、苯醚甲环唑和多福·溴菌腈等6种新型化学药剂对菜豆炭疽病菌的生长抑制作用。结果表明,6种化学药剂对菜豆炭疽病菌均具有抑制作用,其中80%福·福锌可湿性粉剂和10%苯醚甲环唑水分散粒剂的抑菌效果最好,浓度为1 000 mg·L~(-1)时的抑菌率分别达到了77.68%和72.96%,EC_(50)分别为212.41 mg·L~(-1)和296.83 mg·L~(-1)。因此,80%福·福锌可湿性粉剂和10%苯醚甲环唑水分散粒剂可作为田间防治菜豆炭疽病的候选药剂。  相似文献