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LIU An-min LUO Ming LI Guo CAI Wang-qing LI Fang-cheng DENG Yue-fei POON Wai-sang 《园艺学报》2008,24(12):2450-2454
AIM: To construct a lentiviral vector encoding human hepatocyte growth factor (hHGF) for transfection,and to observe the expression of hHGF in human umbilical cord mesenchymal stem cells.METHODS: pUC-SRα/hHGF was subcloned into the expression vector pWPI to construct recombinant pWPI-hHGF.hHGF was identified by gene sequence.Recombinant lentivirus was produced by pWPI-hHGF,pAX2 and pMD2G altogether transient transfection into 293T cells using calcium phosphate method.The pWPI-hHGF and the contructed pWPI-GFP were transfected into human umbilical cord mesenchymal stem cells by the Lipofectamin 2000.Through counting by the fluorescent microscope,the efficiency of the transfection was identified.The expressions of hHGF and GFP in human umbilical cord mesenchymal stem cells were also detected.The concentration of hHGF in cell culture medium was determined by enzyme linked immunosorbent assay (ELISA).RESULTS: DNA sequence showed that hHGF cDNA was correctly inserted into pWPI vector.The positive rate of hHGF transfecting 293T cells was 100 %.Bright green fluorescence in the transfected cells was observed under the fluorescent microscope after 24 h transfection with lentiviral plasmid pWPI-hHGF-GFP,and the transfection rate reached 80%.The difference was distinct between the pWPI-hHGF group and control group in the secretive level of hHGF by Western blotting and the ELISA (P<0.01).CONCLUSION: The recombinant pWPI-hHGF plasmid was successfully constructed and efficient,stable and ectopic expression of hHGF was accomplished in human umbilical cord mesenchymal stem cells. 相似文献
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Effect of ensiling whole crop oat with lucerne in different ratios on fermentation quality,aerobic stability and in vitro digestibility on the Tibetan plateau 下载免费PDF全文
L. Chen G. Guo X. J. Yuan J. Zhang A. Y. Wen X. H. Sun T. Shao 《Journal of animal physiology and animal nutrition》2017,101(5):e144-e153
The objective of this study was to determine the effect of ensiling different ratios of whole crop oat to lucerne on fermentation quality, aerobic stability and in vitro digestibility of silage on the Tibetan plateau. Four experimental treatments were produced varying in the ratio of forages on a fresh matter (FM) basis: 1) 100% oat (control, dry matter (DM) content: 317 g/kg), 2) 90% oat + 10% lucerne (OL10, DM content: 316 g/kg), 3) 80% oat+ 20% lucerne (OL20, DM content: 317 g/kg) and 4) 70% oat+ 30% lucerne (OL30, DM content: 318 g/kg). All treatments were packed into laboratory‐scale silos and ensiled for 60 days and then subjected to an aerobic stability test for 15 days. Further, the four experimental treatments were incubated in vitro with buffered rumen fluid to study the nutrient digestibility. All silages were well preserved with low pH and NH3‐N contents, and high lactic acid contents and V‐scores (evaluation of silage quality). Increasing the lucerne proportion increased (p < 0.05) crude protein (CP) content of silage, whereas neutral (NDF) and acid (ADF) detergent fibre contents were not affected. Under aerobic conditions, the control silage showed higher (p < 0.05) yeast counts (>105 cfu/g FM) followed by OL10 silage, and OL10 silage improved aerobic stability for 74 h. OL20 and OL30 silages showed fewer (p < 0.05) yeasts (<105 cfu/g FM) and markedly (p < 0.05) improved the aerobic stability (>360 h). After 48‐h incubation, OL30 silage increased (p < 0.05) in vitro dry matter digestibility (IVDMD) and neutral detergent fibre digestibility (IVNDFD) compared with the control silage. These results suggest that replacing oat with lucerne had no unfavourable effects on fermentation quality of silage, but improved CP content, aerobic stability IVDMD and IVNDFD. OL30 silage was the best among the three mixed silages. 相似文献
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银杏幼胚离体培养再生植株的研究 总被引:5,自引:0,他引:5
以佛手银杏幼胚为外植体, 研究了不同大小幼胚在不同培养条件下, 愈伤组织和胚状体的诱导发育情况。结果表明: 大于3 mm的幼胚, 在MK +NAA 1.0 mg·L - 1 +BA 1.0 mg·L - 1培养基上胚状体诱导率最高, 达到53.6% , 最多的一块愈伤组织形成多达38个胚状体。暗培养不利于胚状体的发生。在MK培养基上添加10%椰汁对胚状体的生长发育有很好的促进作用, 有34.5%生长成苗。 相似文献
76.
Yufei Guo Chanjuan Shen Anchun Cheng Mingshu Wang Na Zhang Shun Chen Yi Zhou 《Veterinary microbiology》2009,138(3-4):258-265
Anatid herpesvirus 1 (AHV-1) CH virulent strain was first isolated from an infected duck and it was found that this virus strain could induce cytopathic effect (CPE) in duck embryo fibroblast (DEF). Following AHV-1 infection, DEF showed morphological changes such as cell rounding, improved refractivity and detachment from the culture surface. However, its pathological characteristics were not adequately known. Related studies were performed and the results showed that syncytium formation could be observed as the other type of CPE in AHV-1 infection. Hematoxylin-eosin staining and 4’, 6-diamidino-2-phenylindole (DAPI) staining of infected DEF were each used to visualize the shape and distribution of chromatin within nuclei and nuclear fragmentation was observed. Chromatin condensation and margination, as well as formation of apoptotic bodies were observed by transmission electron microscopy (TEM). DNA ladder formation was detected in AHV-1 infected cells and apoptosis of the infected DEF was also detected by flow cytometry analysis of Annexin V-FITC/PI staining method. Therefore, it was suggested that AHV-1 virulent strain can induce syncytium and apoptosis in DEF. Syncytium formation and apoptosis observed in this study may contribute to the elucidation of AHV-1 pathogenesis. 相似文献
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根据酶联免疫(ELISA)试剂盒的原理与研究现状,从人工抗原合成、单克隆抗体制备、试剂盒的反应条件优化、试剂盒技术参数的研究和试剂盒的组装等方面讨论研究过程中的基本操作及存在的问题,为广大科研工作者提供一些理论参考。 相似文献
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