犬瘟热病毒N蛋白基因融合蛋白原核表达条件的优化与蛋白纯化

[目的]提高犬瘟热病毒N蛋白基因在大肠杆菌中表达可溶性GST融合蛋白的含量.[方法]研究诱导剂(IPTG)的最佳加入时间、诱导剂(IPTG)浓度、诱导时间以及不同温度条件对GST - CDV N融合蛋白表达的影响,获得在大肠杆菌中的最佳的表达条件,然后再在最佳条件下大量的表达,用GST - Sepharose 4B亲和层析柱对GST - CDV N融合蛋白进行纯化,得到最大量的可溶性融合蛋白,并对蛋白含量进行测定.[结果]大肠杆菌BL21 (DE3)转化菌在37℃培养3.5 h(OD600 =1.314)时,加入IPTG使终浓度为1 mmol/L,然后在27℃诱导8h,GST - CDV N融合蛋白可溶性表达量最高,达到0.862 mg/mL.[结论]确定了犬瘟热病毒N蛋白基因的优化表达条件,为犬瘟热病毒分子免疫学诊断和亚单位疫苗的研制奠定了基础.     

Juglone inhibits epithelial-mesenchymal transition of prostate cancer cells by regulating Wnt/β-catenin/Snail signaling pathway

AIM: To investigate the mechanism of juglone on epithelial-mesenchymal transition in prostate cancer cells. METHODS: Human prostate cancer LNCaP cells were divided into control group (without juglone), 12.5 μmol/L juglone group and 25 μmol/L juglone group. LNCaP cells in the latter 2 groups were treated with juglone for 24 h. The invasion ability of the LNCaP cells was detected by Transwell assay. The protein expression of E-cadherin, vimentin, Snail and β-catenin was determined by Western blot. The LNCaP cells were treated with LiCl and juglone in combination for 24 h, and the protein expression of Snail and E-cadherin was detected by Western blot.RESULTS: The results of Trans-well invasion assay showed that the invasion ability in juglone groups was significantly decreased (P<0.01). The protein expression of E-cadherin in the LNCaP cells treated with juglone was increased, and the expression levels of vimentin and β-catenin were reduced (P<0.01). Treatment with LiCl significantly attenuated the inhibitory effect of juglone on Snail expression and subsequent down-regulation of E-cadherin expression. CONCLUSION: Juglone inhibits the epithelial-mesenchymal transition by inhibiting the Wnt/β-catenin/Snail signaling pathway in the LNCaP cells.     

非1B/1R类型小麦雄性不育系KTSP3314A败育的生化机制

为了揭示K型非1B/1R类型小麦不育系雄性败育的生化机制。利用K型非1B/1R类型小麦雄性不育系KTSP3314A,其同型保持系TSP3314B,以K型1B/1R类型小麦雄性不育系K3314A、其同型保持系3314B为对照,分别对其叶片和穗子在不同发育时期的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)的活性和丙二醛(MDA)含量的变化进行测定。结果表明,K型非1B/1R类型小麦不育系中SOD,POD及CAT活性变化与K型1B/1R类型小麦雄性不育系酶活性变化一致,K型非1B/1R类型雄性不育小麦叶片和穗子的超氧化物歧化酶(SOD)后期则较低,过氧化氢酶(CAT)、过氧化物酶(POD)的活性后期较高,丙二醛(MDA)含量后期较高。说明二者的雄性败育生化机制基本一致,但前者比后者败育的时期较晚。且超氧化物歧化酶(SOD)活性与雄性育性有关。     

SMEDWI-2 is a PIWI-like protein that regulates planarian stem cells

We have identified two genes, smedwi-1 and smedwi-2, expressed in the dividing adult stem cells (neoblasts) of the planarian Schmidtea mediterranea. Both genes encode proteins that belong to the Argonaute/PIWI protein family and that share highest homology with those proteins defined by Drosophila PIWI. RNA interference (RNAi) of smedwi-2 blocks regeneration, even though neoblasts are present, irradiation-sensitive, and capable of proliferating in response to wounding; smedwi-2(RNAi) neoblast progeny migrate to sites of cell turnover but, unlike normal cells, fail at replacing aged tissue. We suggest that SMEDWI-2 functions within dividing neoblasts to support the generation of cells that promote regeneration and homeostasis.     

Replication and transmission of porcine circovirus type 2 in mice

Little information is known about infection, replication and transmission of porcine circovirus type 2 (PCV2) in species other than swine. Two sets of animal experiments were carried out to investigate the susceptibility of mice to PCV2 and to study their possible role in maintaining and transmitting the virus. In the first experiment 14 mice were inoculated with PCV2 by the intraperitoneal route with 5 x 10(2) TCID50 of the PCV2-ROM strain (Cadar et al., 2007). In a second experiment 24 mice were divided into two groups (A and B); mice in Group A (n = 18) were inoculated orally with 1 x 10(5) TCID50 PCV2-ROM and mice in Group B (n = 6) were left uninoculated until day 12 post inoculation (p.i.), when they were mixed with Group A. The animals were sacrificed at intervals for postmortem investigation and virus genome detection by polymerase chain reaction (PCR). The PCR results indicated that PCV2 could replicate in mice infected intraperitoneally or by the oral route, and that the virus can be transmitted directly from mouse to mouse.     

Sheep performance under intensive continuous grazing of native grasslands of <Emphasis Type="Italic">Paspalum notatum</Emphasis> and <Emphasis Type="Italic">Axonopus compressus</Emphasis> in the subtropical region of the Highlands of Central Mexico

Liveweight gain was evaluated in tropical Dorper X Pelibuey lambs under intensive continuous grazing of native grasslands dominated by Paspalum notatum (PN) or Axonopus compressus (AC) in the subtropics of Central Mexico. Two trials were undertaken. Trial 1 lasted 12 weeks with 10 lambs (initial weight 18 +/- 2.57 kg, 3 months old) per treatment in 2002, and Trial 2 for 13 weeks with 8 lambs (initial weight 24.0 +/- 2.0 kg, 4 months old) per treatment. Lambs were weighed once per week, and liveweight change was estimated by linear regression over day of the experiment, using individual regression coefficients as unbiased estimates of daily liveweight change; analysed in a random block design. Lambs on Trial 1 gained 0.061 kg/lamb/day on PN and 0.047 kg/lamb/day on AC (P > 0.05) at an overall mean stocking rate of 25 lambs/ha. In Trial 2, liveweight gain was significantly larger in PN (0.060 kg/lamb/day) than on AC (0.043 kg/lamb/day) (P < 0.05), at a mean stocking rate of 21.5 lambs/ha. It is concluded that intensive continuous grazing of native grasslands in the subtropics of the highlands of Central Mexico enables moderate liveweight gains for weaned lambs during the rainy season; with better results in grasslands dominated by Paspalum notatum.     

益生菌发酵棉粕对黄羽肉鸡小肠黏膜形态、血清T淋巴细胞亚群及肠道菌群的影响

本试验旨在研究添加不同比例益生菌发酵棉粕对黄羽肉鸡小肠黏膜形态、血清T淋巴细胞亚群及肠道菌群的影响。选用14日龄健康黄羽肉鸡320只,随机分成4组,每组4个重复,每个重复20只鸡,Ⅰ组为空白对照组,Ⅱ、Ⅲ、Ⅳ组分别添加3%、6%和9%发酵棉粕,研究不同添加量对肉鸡生长前期(14～28日龄)、中期(29～45日龄)、后期(46～65日龄)各阶段小肠黏膜形态,中、后期血清T淋巴细胞亚群及后期肠道菌群的影响。研究结果显示:①与Ⅰ组相比,Ⅲ组黄羽肉鸡前、中、后期的十二指肠绒毛高度极显著提高31.90%、25.17%、49.41%(P<0.01),隐窝深度极显著降低9.42%、11.31%、10.83%(P<0.01),V/C极显著提高45.65%、41.19%、67.84%(P<0.01);空肠绒毛高度极显著提高28.72%、28.38%、27.61%(P<0.01),隐窝深度显著或极显著下降6.65%(P<0.05)、6.18%(P<0.01)、7.37%(P<0.01),V/C极显著提高37.84%、36.69%、37.82%(P<0.01);回肠绒毛高度极显著提高76.64%、40.36%、67.20%(P<0.01) ,隐窝深度显著降低4.09%、6.07%、6.02%(P<0.05),V/C极显著提高83.79%、49.27%、78.26% (P<0.01)。②中、后期Ⅳ组黄羽肉鸡CD4⁺/CD8⁺显著提高42.86%、34.95%(P<0.05)。③随着发酵棉粕添加量的增多,黄羽肉鸡的十二指肠、空肠、回肠中乳酸菌的数量逐渐增多,且差异越来越明显,除Ⅱ组十二指肠和空肠段外,其余各组的各肠段差异均显著或极显著(P<0.05;P<0.01)。大肠杆菌和沙门氏菌的数量逐渐减少,且差异越来越明显,除Ⅱ组十二指肠段外,其余各组各肠段差异均显著或极显著(P<0.05;P<0.01)。综上所述,6%发酵棉粕组在提高十二指肠、空肠、回肠绒毛高度、V/C,降低其隐窝深度方面都优于3%、9%发酵棉粕组,而9%发酵棉粕组在提高CD4⁺/CD8⁺、肠道乳酸菌数量和降低大肠杆菌和沙门氏菌数量方面优于3%、6%发酵棉粕组。     

Leukocytic responses and intestinal mucin dynamics of broilers protected with Enterococcus faecium EF55 and challenged with Salmonella Enteritidis

The protective effect of Enterococcus faecium EF55 in chickens challenged with Salmonella enterica serovar Enteritidis phage type 4 (SE PT4) was assessed. The antibacterial effect on the bacterial microflora in the small intestine in relation to white blood cell count, phenotyping of peripheral blood and intestinal lymphocytes, functional activity of lymphocytes and phagocytes and mucin quantitation were investigated. Day-old chicks (85) were randomly divided into four groups. The probiotic group (EF) and Salmonella + probiotic group (EFSE) received E. faecium EF55 (10⁹ CFU – 3 g/group/day) for 21 days. The Salmonella group (SE) and EFSE group were infected with Salmonella Enteritidis (10⁸ CFU in 0.2 ml PBS) in a single dose per os on day four of the experiment. The control group chicks (C) were fed a commercial diet without added bacteria. Supplementation of EF55 in the diet of the chickens in the EFSE group, challenged with S. Enteritidis, caused the density of the intestinal mucin layer to increase significantly in non-specific regions (duodenum and jejunum), but decrease significantly in target regions (caeca) for S. Enteritidis. Probiotic treatment also appeared to result in a significantly higher number of lymphocytes in peripheral blood and a tendency to increase CD3, CD4, CD8, and IgM positive cells 3 days post-infection with S. Enteritidis. The results demonstrated an antibacterial effect and suggested that EF55 had a moderating effect on intestinal mucin production and leukocytic response in the early phase of S. Enteritidis infection.