Secretoglobin and Transferrin Expression in Bronchoalveolar Lavage Fluid of Horses with Chronic Respiratory Disease

Background

Lower expression of secretoglobin and transferrin has been found in the bronchoalveolar lavage fluid (BALF) of a small number of horses with experimentally induced signs of recurrent airway obstruction (RAO) compared to healthy controls.

Hypothesis/Objectives

Secretoglobin and transferrin BALF expression will be similarly decreased in horses with naturally occurring clinical signs of RAO and in horses with experimentally induced clinical signs of RAO as compared to healthy controls and intermediate in horses with inflammatory airway disease (IAD).

Animals

Recurrent airway obstruction‐affected and control horses were subjected to an experimental hay exposure trial to induce signs of RAO. Client‐owned horses with a presumptive diagnosis of RAO and controls from the same stable environments were recruited.

Methods

Pulmonary function and BALF were evaluated from control and RAO‐affected research horses during an experimental hay exposure trial (n = 5 in each group) and from client‐owned horses (RAO‐affected horses, n = 17; IAD‐affected horses, n = 19; healthy controls, n = 5). The concentrations of secretoglobin and transferrin in BALF were assessed using Western blots.

Results

Naturally occurring and experimentally induced RAO horses had similar decreases in BALF transferrin expression, but secretoglobin expression was most decreased in naturally occurring RAO. Secretoglobin and transferrin expression were both lower in BALF of RAO‐affected horses than in IAD‐affected and control horses.

Conclusions and Clinical Importance

Secretoglobin and transferrin expression is decreased in BALF of RAO‐affected horses after both experimental and natural exposure. Secretoglobin and transferrin likely play clinically relevant roles in the pathophysiology of RAO, and may thus be used as biomarkers of the disease.     

Expression of recombinant triangular bream (Megalobrama terminalis) pre-IGF-I in Escherichia coli

The insulin-like growth factor I (IGF-I) cDNA (GenBank Accession No. AY247412) of triangular bream (Megalobrama terminalis) was expressed for the first time in Escherichia coli. To construct the expression plasmid, the IGF-I cDNA was subcloned into prokaryotic-expressing vector pGEX-4T-1. The E.coli JM109 was transformed with the recombinant plasmid pGEX-4T-1-IGF-I, and the transgene expression was observed after being induced with isopropyl-β-D-thiogalactoside (IPTG). The results of SDS polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting indicated that the recombinant fusion protein had immune activity and the molecular weight was about 47 kDa. The results of SDS-PAGE and thin layer scanning showed that the yield of fusion protein had been enlarged with prolonging time. When the time of induced expression was 1, 2, 3, 4, 5, and 6 h, the expression amount was approximately 1.4, 4.3, 8.1, 11.3, 16.3 and 18.8% of total bacterial protein respectively.     

橡胶树肌动蛋白解聚因子原核表达及纯化

本研究根据HbADF序列设计引物扩增基因的编码区,并将其插入到原核表达载体pET28a上,成功构建重组质粒pET28a-HbADF。将重组质粒转化大肠杆菌BL21(DE3)宿主菌,经1 mmol/L IPTG诱导,获得相对分子量为20 ku的融合蛋白。表达蛋白以可溶和包涵体两种形式存在,通过亲和层析方法纯化可溶蛋白并获得HbADF融合蛋白,用抗HIS标签的单抗对纯化蛋白进行了Western blot鉴定。该结果为进一步研究HbADF蛋白特性及功能奠定基础。     

粤西地区甘蔗、香蕉产业发展现状调研分析

简要分析粤西地区甘蔗和香蕉产业发展现状和存在问题,针对存在问题及其今后的发展重点,提出粤西地区甘蔗、香蕉产业发展重点及对策,以促进粤西地区甘蔗和香蕉产业稳健发展。     

2011年鄂西地区水稻稻瘟病菌遗传多样性的分析

[目的]扩大鄂西地区稻瘟病菌菌库,分析其群体结构.[方法]收集鄂西不同地区2011年间的感病稻秆,分离保存稻秆上的穗颈瘟病菌,通过RAPD、rep-Pot2-PCR、REMAP 3种标记对分离的菌株进行分子标记.[结果]从9个稻秆上共分离得到29个稻瘟病菌.遗传多样性分析结果表明,相似性水平在75％时,2011年供试菌株可分为5个遗传宗谱,宗谱3（7个菌株）和4（19个菌株）为优势宗谱,其余3个为稀有宗谱.[结论] 2011年鄂西地区稻瘟病菌由5个遗传宗谱组成.     

基于无损伤取脂的辽西油松资源开发利用策略

通过对辽西油松资源状况及开发利用难点的分析,得出传统取脂方式对油松林的损伤,会严重破坏辽西生态环境,因此传统方式完全不适合辽西油松资源的开发利用。无损伤取脂开发利用策略,不但可以很好地解决辽西油松开发利用的难点,还可以有力地带动辽西油松荒山育林工程。因此无损伤取脂是辽西油松资源开发利用的最佳策略。     

川西亚高山岷江冷杉林土壤生物肥力特征研究

【目的】以川西亚高山岷江冷杉林为研究对象,研究该区表层土壤微生物种群数量、酶活性以及与之相关的土壤养分季节动态等土壤生物肥力特征。【方法】在川西亚高山岷江冷杉林中设置样地,2011-08中旬在样地上、中、下坡位放置分解袋,分别于2011年9月、11月和2012年3月、5月、7月中旬随机取回部分分解袋,测定其中土壤养分、微生物数量和酶活性,并进行相关分析和主成分分析。【结果】林地土壤微生物数量、酶活性和养分含量季节差异显著,在3种微生物中以细菌数量最多;细菌和真菌数量,蔗糖酶和过氧化氢酶活性,有机质、全氮、全钾、全磷、速效氮和有效磷含量7月最高,放线菌数量、脲酶活性9月最高。相关分析表明,细菌数量与全钾、有机质、全磷、速效氮、有效磷含量呈显著正相关,真菌数量与有机质、全氮、速效钾含量呈显著正相关,放线菌数量与全磷、速效氮、有效磷含量呈显著正相关,脲酶、蔗糖酶、过氧化氢酶活性与有机质、全氮、全钾、速效氮、速效钾含量呈显著正相关;主成分分析表明,第1主成分贡献率为49.766%,主要综合了蔗糖酶、全钾、全磷、真菌的信息。【结论】川西亚高山岷江冷杉林土壤微生物数量、酶活性及土壤养分三者之间相互影响,共同反映土壤生物肥力特征。     

西部欠发达地区公共体育服务体系的现状与对策研究* ——以贵州省为例

以社会成员对公共体育服务的需求为切入点,采用文献资料、访谈、问卷调查和数理统计等研究方法对贵州省公共体育服务的基础设施类、信息服务类、要素保障类进行调查分析,探讨西部欠发达地区公共体育服务体系的构建。     

血清1和3型多杀性巴氏杆菌铁调控外膜蛋白共同抗原的初步分析

用普通BHI和缺铁BHI培养基培养禽多杀性巴氏杆菌C48-19（血清1型）、P1059株（血清3型）,提取外膜蛋白（OMPs）,SDS-PAGE电泳比较不同培养条件下两株细菌的外膜蛋白,发现缺铁培养时2株细菌均增加了87、81、79、64、60．5ku铁调控外膜蛋白（IROMPs）。以C48-19株攻毒后耐过的鸡血清作为抗体进行免疫印迹,检测到其中的87、79和60．5ku蛋白是2株细菌共同的、具有交叉免疫反应性的IROMPs,推测这3种蛋白可能是这2个菌株的共同抗原,并可能在C48-19感染的鸡体内特异表达;对鸡胚尿囊液中培养的C48-19株OMPs的SDS-PAGE电泳和免疫印迹试验,也观察到类似IROMPs的表达。这些IROMPs的发现为亚单位疫苗的研制提供了候选蛋白。     

犬瘟热病毒H基因的序列分析与表达

以临床疑似犬瘟热病犬外周血总RNA为模板,采用RT-PCR方法扩增犬瘟热病毒(CDV)血凝素(H)基因。序列测定和分析表明该病料犬瘟热病毒H基因序列与国内其他地区分离到的毒株同源性较高(94.6%~99.2%),而与Onderstepoort株、Convac株等疫苗毒株的同源性较低(90.4%~91.2%)。分子遗传进化分析显示所有毒株可分为7个大的分支,且各分支间具有一定的地域性,其中待检病料中CDV与大多数中国分离株一样处于Ⅰ型。进一步以H全基因为模板,截短表达其3′端816 bp、459 bp两个片段,并克隆入原核表达载体pET30 a进行表达。结果显示它们分别能表达大小约为36.4 ku和22.2 ku的融合蛋白。免疫转印表明该纯化蛋白均可与犬瘟热病毒抗血清发生阳性反应,说明重组蛋白具备抗原性,可用于进一步的血清学研究。