H5N1亚型禽流感病毒A/Guangxi/1/2005株抗药机制的研究

为研究H5N1亚型禽流感病毒(AIV)的抗药机制,本研究选取Clade2.3.4亚群中一株对金刚烷胺敏感的人源AIV A/Guangxi/1/2005(H5N1)(S-GX05),用抗流感病毒药物金刚烷胺对其进行定向诱导,筛选出一株抗药性病毒株,命名为R-A/Guangxi/1/2005(R-GX05)。通过全基因测序并与S-GX05全基因序列进行对比,结果显示S-GX05只在其M2蛋白中有一个氨基酸位点发生突变,即A30P;抗药性鉴定这两株病毒的半数药物抑制浓度(IC50)分别为0.9μM和48.9μM,表明R-GX05对金刚烷胺表现出一定程度的抗性。动物实验证实,这两株病毒对BALB/c小鼠的致病性基本一致,均表现出高致病性,其MLD50分别为4.7 log10 EID50和5.0 log10 EID50,两株病毒在小鼠体内各组织脏器中的分布及增殖能力也基本相同。这些结果表明,S-GX05在药物压力下产生抗药性后,并未引起其它生物学特性的改变。A30P的发现为进一步从分子水平上研究H5N1亚型AIV的抗药机制及新型抗流感新药的研发奠定了基础。     

重新认识农区山羊放牧饲养

文章从山羊养殖历史、生物学特性、自然社会、经济效益、生态环保、市场消费及国外养殖进展等方面进行研究,认为我国南方农区山羊饲养应回归放牧养羊,可以实现社会生态经济的可持续发展。同时提出放牧养殖的标准化建设思路。     

A dominant antigenic epitope on SARS-CoV spike protein identified by an avian single-chain variable fragment (scFv)-expressing phage

Severe acute respiratory syndrome (SARS) is a newly emergent human disease, which requires rapid diagnosis and effective therapy. Among antibody sources, immunoglobulin Y (IgY) is the major antibody found in chicken eggs and can be used as an alternative to mammalian antibodies normally used in research and immunotherapy. In this study, phage-expressing chicken monoclonal scFv antibody was chosen and characterized with phage display antibody technology. Truncated fragments of SARS-CoV spike protein were cloned in pET-21 vector and expressed in BL-21 Escherichia coli (E. coli) cells. After purification, the purity of these recombinant spike proteins was examined on SDS-PAGE and their identity verified with Western blot analysis using anti-his antibodies and sera from convalescent stage SARS-CoV-infected patients. Using these bacteria-derived proteins to immunize chickens, it was found that polyclonal IgY antibodies in the egg yolk and sera were highly reactive to the immunogens, as shown by Western blot and immunocytochemical staining analysis. A phage displaying scFv library was also established from spleen B cells of immunized chicken with 5 x 10(7) clones. After four panning cycles, the eluted phage titer showed a 10-fold increase. In sequence analysis with chicken germline gene, five phage clones reacted, with large dissimilarities of between 31 and 62%, in the complementarity-determining regions, one dominant phage 4S1 had strong binding to fragment Se-e, located between amino acid residues 456-650 of the spike protein and this particular phage had significantly strong binding to SARS-CoV-infected Vero E6 cells. Based on the results, we conclude that generating specific scFv-expressing phage binders with the phage display system can be successfully achieved and that this knowledge can be applied in clinical or academic research.     

不同的干燥工艺对荷花蜂花粉功能成分的影响

探讨了5种干燥工艺对荷花蜂花粉功能成分的影响。微波干燥的干燥效率最高,60%火力处理540s可使水分从31.41%降到6.42%;干燥方式对荷花蜂花粉蛋白质影响较小;微波干燥后总黄酮提取率最高,为13.20mg/g;真空冷冻干燥后维生素C和过氧化氢酶含量最高,分别为48.89mg/100g和29.86mg/g·min。总的来说,真空冷冻干燥对荷花蜂花粉的功能成分影响最小;常压恒温干燥对功能成分影响较小、工艺简单、对设备要求低,宜于大规模应用于实际生产中。     

舍饲和田羊与卡拉库尔羊生长发育指标的测定

本研究旨在对新疆南部地方品种绵羊平原型和田羊、山区型和田羊及卡拉库尔羊的17种生长发育指标进行检测分析,了解农区舍饲条件下新疆南部地方品种绵羊的基本生长发育情况。主要进行了2个品种、品系绵羊断奶后7个多月期间的17项生长指标的检测,通过SPSS 17.0统计软件分析其各项指标的差异显著性。结果表明：6月龄时,平原型和田羊与山区型和田羊间体高、尻长存在差异显著性(P<0.05),而后胸宽、尾长、尾宽、尾厚的差异极显著(P<0.01);山区型和田羊与卡拉库尔羊间前胸宽、后胸宽存在差异显著性(P<0.05);平原型和田羊与卡拉库尔羊间体重、胸围存在差异显著性(P<0.05),且前胸宽、后胸宽、尾长、尾宽、尾厚的差异极显著(P<0.01)。所检测各项指标的平均月增长量,平原型和田羊与山区型和田羊间体高存在差异显著性(P<0.05),而山区型和田羊与卡拉库尔羊间体高差异极显著(P<0.01);山区型和田羊与卡拉库尔羊管围存在差异显著性（P<0.05）;平原型和田羊与卡拉库尔羊间尾厚存在差异显著性(P<0.05)。对于绵羊体重、体长、体高、胸围等生长发育规律的研究结果表明,平原型和田羊的生长发育与山区型和田羊以及卡拉库尔羊相比较在这些方面具有优势。     

重要营养素--血红素铁

血红素铁是卟啉类化合物,是哺乳动物血液中的天然色素和生物铁,广泛应用于医药、食品和化工业。文章就血红素铁的理化特性、生理功能及应用进行了简述,并对其常用提取方法进行了比较概述。     

猪毛滴虫病病原及诊治的研究进展

猪毛滴虫病的历史较短,关于该病的研究和病例报道也较少。近年来,随着该病导致保育猪腹泻病例日益增多,猪毛滴虫病对养猪业的危害日益显现。本文就该病的病原、危害性、致病机理、流行病学、临床症状、病理变化、诊断以及防治作一综述,旨在为猪毛滴虫病的研究和防控提供参考。     

益生菌发酵饲料对猪的生长和肉质的影响

应用益生菌发酵饲料喂猪,能提高饲料中粗蛋白含量,明显降低猪粪臭味,促进生长、降低料肉比和提高猪肉品质。     

传统发酵食品来源植物乳杆菌的发酵特性及其在酸奶中的应用

对传统发酵食品来源的三株植物乳杆菌(Lactobacillus plantarum)的发酵特性进行试验分析。结果表明植物乳杆菌DH1-XHG-3具有适度的菌体自溶特性和蛋白水解活力,且能够高产胞外多糖(99.7mg/L)。将植物乳杆菌DH1-XHG-3作为附属发酵剂应用于酸奶中,发现添加植物乳杆菌的酸奶样品经冷藏21d后菌株DH1-XHG-3的活菌数仍保持在108 CFU/m L,且酸奶样品的乳清析出率低于对照组,感官评定、质构特性和持水力均高于对照组。以上结果证明植物乳杆菌DH1-XHG-3适合作为附属发酵剂应用于发酵乳制品中。     

Gene cloning of porcine adiponectin gene from adipose tissue and construction of its eukaryotic expression vector

To clone adiponectin (ADPN) gene from Shaziling porcine adipocyte and construct its eukaryotic expression vector, total RNA was extracted from subcutaneous fatty tissue. One pair of specific primers was designed by Primer 5.0 software according to the sequence of ADPN gene of porcine available in GenBank. The ADPN gene was amplified by PCR from cDNA and cloned into pMD18‐T vector to construct recombinant clonal vector pMD‐ADPN, sequenced and analysed. A recombinant expression plasmid pPICZaA‐ADPN was constructed by subcloning the cloned ADPN gene into the linearized pPICZaA vector. Then, the plasmid pPICZaA‐ADPN was expressed in Pichia pastoris (GS115) by electrotransformation. Western blot and Bradford analysis were used to determine the target protein induced by methanol. Results showed that the genome size of ADPN was 732 bp and encoded 244 amino acid, the nucleotide sequence of ADPN shared 100% identity with that of porcine available in GenBank. Western blot and Bradford analysis showed that the recombinant ADPN was expressed in GS115 correctly and has certain immune activity. The expression level of ADPN was 28.5 μg/ml. In conclusion, the recombinant ADPN could express in eukaryotic expression vector pPICZaA‐ADPN constructed in this study effectively.