High level expression of recombinated hepatocyte growth factor in human umbilical cord mesenchymal stem cells

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.     

Effect of ensiling whole crop oat with lucerne in different ratios on fermentation quality,aerobic stability and in vitro digestibility on the Tibetan plateau

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.     

银杏幼胚离体培养再生植株的研究

 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。     

Anatid herpesvirus 1 CH virulent strain induces syncytium and apoptosis in duck embryo fibroblast cultures

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.     

饲料中霉菌毒素ELISA试剂盒制作的研究进展

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。     

土壤湿度对三氮苯类除草剂药效的影响及优化方案

以稗草为生物测定材料,运用二次正交旋转组合设计,以土壤湿度和除草剂用量二因子为决策变量,对稗草的抑制率为目标函数,研究土壤湿度对三氮苯类除草剂药效的影响。结果表明,适当的土壤水分是三氮苯类除草剂发挥药效的重要因素,药效随土壤湿度的提高而提高。不同的土壤湿度对不同除草剂药效影响各异,高湿条件下,湿度差异对药效影响大小依次为嗪草酮、西草净、扑草净、莠去津,低湿条件下则相反。除草剂用量与土壤湿度存在最佳发挥药效的组合。     

Effects of Compound Plant Nutrients on Rumen Fermentation Parameters,Rumen Microbiota and Fatty Acid Composition of Longissimus dorsi Muscle in Finishing Small Tail Han Sheep北大核心CSCD

This study was conducted to investigate the effects of compound p1ant nutrients (CPN) on rumen fermentation parameters, rumen microbiota and fatty acid composition of longissimus dorsi musc1e in finishing sma11 tai1 Han sheep. Sixteen 4-month-o1d finishing sma11 tai1 Han sheep with an initia1 body weight (BW) of (24.18±0.31) kg were random1y divided into two groups, name1y, with 8 rep1icates per group and 1 sheep per rep1icate. The sheep in the contro1 group were fed a basa1 diet, whereas the sheep of the contro1 group (CPN group) was fed the basa1 diet supp1ementation with 3‰ CPN. The experiment 1asted for 97 days after 7 days adaption. The resu1ts showed as fo11ows: compared with the contro1 group, 1) adding CPN decreased the concentration of ammonia nitrogen (NH3-N) (P<0.05); 2) CPN supp1ementation affected beta diversity of rumen microbiota; 3) the re1ative abundance of Acidobacteriota, Erysipe1otrichaceae_UCG-002, Lactobacillus, and Megasphaera were enhanced by adding CPN (P<0.05), whereas, the re1ative abundance of Bacteroidetes and Prevotella was 1ower (P<0.05); 4) the supp1ementation of CPN had no significant effect on fatty acid composition of longissimus dorsi musc1e of fattening sma11-tai1ed Han sheep (P>0.05); 5) the contents of tota1 MUFA, C18:1n9c, C14:1, C16:1, C18:2n6c, C18:3n3 and n-3PUFA in the longissimus dorsi musc1e were corre1ated with the re1ative abundance of Megasphaera, Erysipe1otrichaceae_UCG-002, Succiniclasticum and Ruminococcus (P<0.05). In conc1usion, CPN can regu1ate the rumen microbiota structure and reduce the rumen NH3-N concentration of fattening sma11-tai1ed Han sheep. In production practice, CPN can be used as a rumen eco1ogica1 regu1ator. © 2023 Authors. All rights reserved.     

海产品中副溶血弧菌的分离与鉴定

副溶血弧菌（Vibrio Paraaemolyticus)是一种致病性嗜盐菌，常引起人类食物中毒，通过对四种不同海产品的检测，发现该菌检出率很高，其中，海虾中的检出率高达90％，海产鱼为70％，贝类为60％，海蟹为40％，平均检出率为65％，动物试验证实，该菌致病性较强。