鸡病毒性关节炎病毒C-98分离株S1基因的克隆与序列分析

提取鸡病毒性关节炎病毒（AVAV）内蒙古分离株（C-98株）总RNA，参考GenBank中禽呼肠孤病毒（ARV）S1133株S1基因序列设计了2对引物，应用RT—PCR技术扩增了病毒S1基因，将S1基因cDNA克隆到pGEM—T Easy载体后测序。将测定序列拼接后与参考毒株ARV-176、ARV-138、ARV-S1133、MDRV—YJL的S1基因序列进行了比较。结果显示，AVAV C-98分离株的S1基因与强毒株ARV-176株的同源性最高，达99．9％；与MDRV—YJL的同源性为99．3％，与弱毒疫苗株ARV—S1133的同源性也达97．9％；与ARV-138株的同源性最低，为81．2％。表明，AVAVC-98株与参考毒株的差异较小，与疫苗株ARV-S1133有很高的同源性。     

In vitro effect of recombinant human tumor necrosis factor-alpha on canine neutrophil apoptosis

Neutrophils (polymorphonuclear leukocytes: PMNs) are essential for the host defense against various infections and are often injurious to the host, causing inflammatory diseases where tumor necrosis factor-alpha (TNF-alpha) is suggested to play an important role. Since an effect of TNF-alpha on canine PMN apoptosis has not been studied, canine PMNs were stimulated with recombinant human (rh)TNF-alpha in the present study to investigate the effect of TNF-alpha on canine PMN apoptosis. PMN apoptosis and function to produce ROS were assessed by flow cytometry. Delayed apoptosis was observed in the PMNs treated with rhTNF-alpha at 100 ng/ml, accompanied by retention of capability to produce ROS. However, PMN apoptosis was accelerated by rhTNF-alpha combined with cycloheximide. Therefore, it is indicated that TNF-alpha is able to activate anti- and pro-apoptotic pathways in PMNs and that the inhibition of PMN apoptosis by TNF-alpha requires protein synthesis in the PMNs.     

The effect of Epigallocatechin‐3‐gallate on small intestinal morphology,antioxidant capacity and anti‐inflammatory effect in heat‐stressed broilers

This study was to investigate the effects of Epigallocatechin‐3‐gallate (EGCG) on intestinal morphology, antioxidant capacity and anti‐inflammatory response in heat‐stressed broiler. A total of 192 2‐week‐old Arbour Acres broilers chickens were divided into four groups with six replicates per group and eight chickens per replicate: one thermoneutral control group (28°C, group TN), which was fed the basal diet; and three cyclic high‐temperature groups (35°C from 7:00 to 19:00 hr; 28°C from 19:00 hr to 7:00 hr, heat stress group), which were fed the basal diet supplementation with EGCG 0 mg/kg (group HS0), 300 mg/kg (group HS300) and 600 mg/kg (group HS600). The gut morphology and intestinal mucosal oxidative stress indicators, as well as intestinal barrier‐related gene expression, were analysed. The results showed that compared with group TN, heat stress reduced the villus height (VH), activities of glutathione peroxidase (GSH‐Px), superoxide dismutase (SOD)and catalase (CAT), increased the crypt depth (CD) and malondialdehyde (MDA)content at 21, 28 and 35 days (p < 0.05). After the heat‐stressed broilers were supplemented with EGCG, VH, VH/CD (V/C), and the activities of GSH‐Px, SOD and CAT were increased, and CD and MDA content were reduced compared with those in group HS0 without EGCG supplementation at 21, 28 and 35 days (p < 0.05). The EGCG supplementation promoted the gene expression of nuclear factor‐erythroid 2‐related factor 2 (Nrf2), Claudin‐1, Mucin 2 (Muc2) and alleviated the nuclear factor‐kappa B (NF‐κB) and lipopolysaccharide‐induced tumour necrosis factor (LITAF) gene expression compared with group HS0 (p < 0.05). Moreover, intestinal morphology was strongly correlated with antioxidant ability and inflammatory response. In conclusion, EGCG alleviated the gut oxidative injury of heat‐stressed broilers by enhancing antioxidant capacity and inhibiting inflammatory response.     

禽呼肠孤病毒σB和σC蛋白在昆虫细胞中的共表达

本研究旨在获得具有天然构象和活性良好的禽呼肠孤病毒（ARV）σB和σC蛋白,采用杆状病毒表达系统在昆虫细胞中共表达σB和σC蛋白,并对其活性进行鉴定。根据NCBI数据库中ARV σB和σC基因序列设计引物,将两个基因克隆至pFast-Dual载体,转化大肠杆菌DH10Bac感受态细胞,筛选获得重组穿梭杆粒。通过转染sf9细胞,筛选到含有σB和σC基因的重组病毒。将重组病毒感染sf9细胞,通过优化接毒量和收获时间等参数,确定最佳表达条件,在sf9细胞中高效共表达σB和σC蛋白。Western blotting和间接免疫荧光（IFA）检测结果表明,成功在sf9细胞中共表达了ARV σB和σC蛋白,并具有很好的生物学活性。本试验结果为开发鉴别诊断试剂以及ARV颗粒疫苗的研究奠定了基础。     

山西铁杆蒿草地群落碳、氮密度区域差异及其驱动因素

为明确山西铁杆蒿（Artemisia sacrorum）群落碳、氮密度的区域差异,以及导致这种空间变化的驱动因素,本试验以山西省铁杆蒿草地群落为研究对象,通过测定晋北半干旱地区和晋南半湿润地区铁杆蒿群落的碳（C）、氮（N）密度,分析了其与年均气温、年降水量、土壤理化性质和地上、地下生物量等生态因子的相关性。结果表明：山西南北铁杆蒿群落碳、氮密度空间分布存在差异,晋南地区铁杆蒿群落的群落碳、氮密度分别为651.83 g C·m^-2和20.63 g N·m^-2,晋北地区铁杆蒿群落的群落碳、氮密度分别为418.89 g C·m^-2和13.90 g N·m^-2。以群落各组分碳、氮密度代表样点进行RDA（Redundancy analysis）冗余分析排序得出造成两地区群落碳、氮密度分异的主要生态因子均为地上生物量、地下生物量、年均气温、年降水量、土壤有机碳含量以及土壤砾石比。可见,山西晋南和晋北地区铁杆蒿群落碳、氮密度空间分布存在差异,不同气候区铁杆蒿群落碳、氮密度空间差异与生物量的积累密切相关,驱动晋南和晋北地区铁杆蒿群落碳氮密度分异的环境因素为年均气温、年降水量、土壤有机碳含量以及土壤砾石比。     

维生素K_3对五龙鹅生长性能、屠宰性能及养分表观利用率的影响

本试验旨在研究饲粮中维生素K3添加水平对五龙鹅生长性能、屠宰性能及养分表观利用率的影响。试验分为2个阶段。1~4周龄阶段,选用1日龄五龙鹅360只,随机分为6个组,每个组6个重复,每个重复10只鹅。Ⅰ组为对照组,饲喂基础饲粮(维生素K3含量1.23 mg/kg),Ⅱ~Ⅵ组在基础饲粮中分别添加1、2、4、8、16 mg/kg的维生素K3。5~16周龄阶段,选用28日龄五龙鹅324只,随机分为6个组,每个组6个重复,每个重复9只鹅。A组为对照组,饲喂基础饲粮(维生素K3含量1.18 mg/kg),B~F组在基础饲粮中分别添加2、4、8、16、32 mg/kg的维生素K3。试验期16周。结果表明:1)经回归分析得出,1~4周龄阶段五龙鹅饲粮中添加4.81 mg/kg维生素K3,平均日增重最大;5~16周龄阶段饲粮中添加11.59 mg/kg维生素K3,平均日增重最大。2)与对照组相比,1~4周龄阶段,饲粮中添加4 mg/kg维生素K3能显著或极显著提高鹅的全净膛率和腹脂率(P0.05或P0.01);5~16周龄阶段,饲粮中添加8 mg/kg维生素K3能显著或极显著提高屠宰率、半净膛率、全净膛率和腿肌率(P0.05或P0.01)。3)与对照组相比,5~16周龄阶段,饲粮中添加8 mg/kg维生素K3能极显著提高钙的表观利用率(P0.01),显著提高干物质、粗蛋白质、粗脂肪、中性洗涤纤维、酸性洗涤纤维和磷的表观利用率(P0.05)。由此可见,本试验条件下,1~4周龄和5~16周龄五龙鹅饲粮中维生素K3适宜添加水平分别为4.81和11.59 mg/kg。     

Basic characterization of avian β‐defensin genes in the Japanese quail,Coturnix japonica

In this study, we identified a cluster of 14 avian β‐defensins (AvBD; approximately 66 kbp) in the Japanese quail, Coturnix japonica. Except for AvBD12 (CjAvBD12) and ‐13, the CjAvBDs coding sequences exhibited greater than 78.0% similarity to the respective orthologous chicken AvBD genes (GgAvBD). The putative amino acid sequence encoded by each CjAvBD contained six cysteine residues and the GXC (X1‐2) motif considered essential for the β‐defensin family. Each CjAvBDs also formed a sub‐group with the respective orthologous genes of various bird species in a phylogenetic tree analysis. Synteny between the CjAvBD cluster and GgAvBD cluster was confirmed. The CjAvBD cluster was mapped on the long‐arm end of chromosome 3 by linkage analysis based on single nucleotide polymorphisms (SNPs) of CjAvBD1 and CjAvBD12 (approximately 46kbp), as well as GgAvBD cluster. We also confirmed that CjAvBD1, ‐4, ‐5, ‐9, and ‐10 are transcribed in 20 tissues, including immune and digestive tissues. However, our experimental data indicated that the CjAvBD cluster lacks the AvBD3 and ‐7 loci, whereas the CjAvBD101α, ‐101β, and ‐101θ loci arose from gene duplication of the AvBD6 orthologous locus in the CjAvBD cluster after differentiation between Coturnix ‐ Gallus.     

袋鼠摩根氏菌生物特性鉴定及系统发育分析

本研究利用Biolog快速鉴定系统、16S rRNA序列分析及传统细菌鉴定方法对3株分离自北京动物园袋鼠肺脏的菌株进行形态学、培养特性、生化特性、小鼠致病性等生物特性的鉴定及系统发育分析。结果表明,3株菌株均为摩根氏菌,对昆明小鼠有强致病性;3株袋鼠摩根氏菌16S rRNA序列与LMG7874模式株同源性均为99.8%,且位于系统发育树的同一分支。     

Glucosinolate profiling of seeds and sprouts of B. oleracea varieties used for food

Consumption of plants of the species Brassica oleracea is related to a decreased incidence of certain cancer forms in humans, and this has been linked to the presence of glucosinolates in those vegetables. After ripe seeds, sprouts of some brassicaceous plants contain the highest concentration of these compounds and are therefore a good source of glucosinolates for chemoprotection. In the present experiments, the content and distribution of glucosinolates was determined in ripe seeds and sprouts (seedlings) of five varieties of B. oleracea (white cabbage, red cabbage, Savoy cabbage, broccoli and cauliflower) by high performance liquid chromatography. The type and concentration of individual glucosinolates varied according to variety of B. oleracea, plant parts in which they occurred and the sprouting period of the seed. Concentration of alkyl glucosinolates decreased whereas that of indol-3-ylmethylglucosinolates increased throughout the sprouting period. Roots had the highest glucosinolate concentration in four and seven day old sprouts whereas at both sprouting times, cotyledons had the highest concentration of alkylthio- and alkylsulphinylglucosinolates.     

Effect of acupuncture on CD4+ CD25+ Foxp3+ regulatory T-cells in rats of embryo implantation failure

AIM：To observe the effect of acupuncture on CD4+ CD25+ Foxp3(forkhead box P3)+ regulatory T-cells(Treg cells) in rats with embryo implantation failure. METHODS：One hundred and forty-four pregnant rats were randomly divided into control group(N), mifepristone treatment group(M), mifepristone+acupuncture treatment group(A) and mifepristone+progestin treatment group(W). The rats in groups M, A and W were treated with mifepristone-sesame oil solution on day 1, while the rats in group N were injected with the same amount of sesame oil. The Housanli(ST36) and Sanyinjiao(SP6) points were selected for acupuncture. From day 1 to the time of death, the rats in group A were fasten up and then the acupuncture was performed. Accordingly, the rats in group N and group M were only fixed, and the rats in group W were given progestin. Implanted embryos in each group were counted. The proportions of CD4+ CD25+ Foxp3+ Treg cells in peripheral blood and CD4+ Foxp3+ Treg cells in the endometrium were detected by flow cytometry. The mRNA and protein levels of Foxp3 were determined by real-time PCR and Western blotting, respectively. RESULTS：Compared with group N, the number of implanted embryos, the percentages of CD4+ CD25+ Foxp3+ Treg cells in peripheral blood and CD4+ Foxp3+ Treg cells in the endometrium, and the expression of Foxp3 protein and mRNA in the endometrium were significantly decreased in group M(P<005). Compared with group M, the above indexes in group A and group W were significantly increased. CONCLUSION: The effect of acupuncture in rats with embryo implantation failure may be closely correlated with the modulation of CD4+ CD25+ Foxp3+ Treg cells.