目的：蓝氏贾第鞭毛虫作为一种重要的人兽共患寄生虫病贾第虫病的病原，其对SPF实验动物质量造成的潜在威胁不容忽视。本研究的目的是建立蓝氏贾第鞭毛虫快速诊断方法，并对17个生产厂家516批2562只SPF实验动物检查结果进行分析。方法用直接镜检法、快速姬姆萨染色法和多重PCR方法，对蓝氏贾第鞭毛虫进行检测。结果在SPF实验动物中检出数量众多的蓝氏贾第鞭毛虫滋养体和包囊，鉴定出蓝氏贾第鞭毛虫18S rDNA、β－giardin、TPI和GDH基因。直接镜检法、快速姬姆萨染色法和多重PCR方法均能检出蓝氏贾第鞭毛虫。17个生产厂家516批2562只SPF实验动物蓝氏贾第鞭毛虫阳性检出率22．9％（586／2562）。结论直接镜检法、快速姬姆萨染色法和多重PCR方法具有高度的敏感性和特异性，可用于蓝氏贾第鞭毛虫的快速诊断。17个生产厂家516批SPF实验动物蓝氏贾第鞭毛虫检查结果未能全部符合规定。

Objective Giardia lamblia is an important pathogen of zoonosis giardiasis , it poses a potential threat to the quality of SPF (specific pathogen-free) laboratory animals cannot be ignored.The aim of this study is to establish the method of rapid diagnosis of Giardia lamblia, and analyze the test results of 516 batches form 17 manufactures.Methods Direct microscopy, Giemsa-fast staining and multiplex polymerase chain reaction (multiplex PCR) were applied to detect Giardia lamblia.Results Numerous of Giardia lamblia trophozoites and cysts were detected in SPF laboratory animals by using direct microscopy and Giemsa-fast staining, and multiplex PCR were performed to identify 18S rDNA,β-giardin, TPI and GDH genes of DNA extracted from these trophozoites and cysts identified Giardia lamblia.Direct microscopy, Giemsa-fast staining, and multiplex PCR methods can be used to detect Giardia lamblia.Of the 2562 SPF laboratory animals studied, 22.9%(586/2562) were positive for Giardia l

明确鞭毛蛋白的黏膜佐剂作用,并研究其与DnaJ蛋白联合免疫对肺炎链球菌感染的保护作用。方法:含有重组质粒pET-28(a)/鞭毛蛋白基因的大肠杆菌DE3经IPTG诱导后表达鞭毛蛋白,纯化后用于后期动物实验如下:将C57BL/6小鼠随机分为四组通过鼻腔分别滴入鞭毛蛋白(10μg)与DnaJ蛋白(10μg)混合剂(实验组)、DnaJ蛋白(10μg)(对照组1)、DnaJ蛋白(10μg)与GST蛋白(10μg)混合剂(对照组2),每组均30μl。通过检测抗体的效价、亚型以及细胞因子水平来评估鞭毛蛋白能否提高肺炎链球菌DnaJ蛋白诱导的免疫效应;然后用肺炎链球菌D39(5×107CFU)进行鼻腔攻毒,观察小鼠的生存率,以评估其保护效应。结果:实验组较对照组产生更高效价的血清IgG(其中主要产生IgG1)和更高水平的细胞因子IFN-γ、IL-17A和IL-4;另一方面实验组对致死性D39感染的保护率达60%,DnaJ蛋白免疫组的保护率为50%。结论:鞭毛蛋白与DnaJ蛋白联合应用可以增强宿主对DnaJ蛋白的免疫反应,并且对致死剂量的肺炎链球菌感染有保护作用,提示鞭毛蛋白可以作为肺炎链球菌DnaJ蛋白疫苗的佐剂应用。

To make sure the role of flagellin as mucosal adjuvants and the protective effect of streptococcus pneumonia infection when combined with DnaJ protein.Methods:Recombinant plasmid pET-28 ( a)/flagella was transferred to E.coli BL21(DE3).Over-expression of flagella was induced by IPTG and purification for animal study.All the C57BL/6 mice were randomly divided into three groups ,then respectively intranasally given the mixture of flagellin and DnaJ protein ( experimental group ) , DnaJ protein( control group 1 ) and the mixture of GST and DnaJ protein ( control group 2 ).The serum IgG and its subtype , cytokines secreted by mice spleen cells were detected by ELISA.At last all the C57BL/6 mice were intranasally immunized with Streptococcus pneumoniaD39.The protective effect by survival times were evaluated.Results: The mice of experimental group could secrete high level of serum IgG and cytokines IFN-γ,IL-4 and IL-17A.What more,the survival rate of mice in experimental group wa

目的：对从新疆实验动物研究中心饲养的封闭群灰仓鼠体内分离到的1株鞭毛虫进行形态学鉴定及基因鉴定。方法取灰仓鼠回盲部内容物进行直接涂片和常规姬姆萨染色后镜检观察，提取虫体总DNA，PCR扩增该鞭毛虫的16S rRNA基因，测序后与国外已报道的鞭毛虫进行核酸同源性分析，并应用MEGA5.22软件绘制系统发育进化树。结果形态学观察表明分离到的鞭毛虫为鼠三毛滴虫。测序后核酸同源性分析表明鼠三毛滴虫新疆灰仓鼠分离株16S rRNA序列与国外已报道的三毛滴虫高度同源。系统发育进化树表明鼠三毛滴虫新疆灰仓鼠分离株序列与已报道的鼠三毛滴虫16S rRNA（序列号AY886846.1）位于同一进化分支，与其他相关三毛滴虫亲缘关系较远。结论形态学鉴定和16 S rRNA基因分析表明，此次从新疆实验动物研究中心饲养的封闭群灰仓鼠体内分离到的鞭毛虫为鼠三毛滴虫。

Objective To conduct morphological observation and gene identification of the strain of flagellate iso -lated from Cricetulus migratorius in the Xinjiang Research Center for Experimental Animals .Methods The ileocecal con-tents of C.migratorius were microscopically examined on direct smear with Wright-Giemsa staining , and the total RNA iso-lated from Xinjiang C.migratorius was extracted and 16S rRNA was amplified by PCR , and then sequenced .Furthermore the homology was compared and the phylogenetic tree was developed using MEGA 5.22 software.Results Morphological observation indicated that the isolated flagellate was Tritrichomonas muris.The 16S rRNA gene sequence of the Xinjiang C. migratorius isolate shared highly homology with that of other Tritrichomonas.Phylogenetic tree analysis indicated that the 16S rRNA gene of Xinjiang C.migratorius isolate was classified into a subgroup with T.muris 16S rRNA (U85966.1), but was relatively distant relative from other related tritrichomonas.

背景与目的已有的研究表明：Toll样受体5（toll-likereceptor5,TLR5）在肿瘤起始和发展中发挥重要作用。我们前期研究发现，TLR5在非小细胞肺癌（non-smallcelllungcancer,NSCLC）组织中高表达，但其在NSCLC高表达后的信号通路活化情况的研究并不多见。本研究旨在探讨TLR5在不同NSCLC细胞株上的表达，及其在NSCLC细胞中活化的机制。方法用免疫荧光、RT-PCR和Westernblot方法检测TLR5在三种不同NSCLC细胞株中的表达。分别用0μg/mL、0.01μg/mL、0.1μg/mL、1μg/mL、5μg/mL、10μg/mL的鞭毛蛋白刺激，用NF-κB荧光素酶报告基因质粒瞬时转染后，检测细胞内NF-κB荧光素酶的活性。选择TLR5表达最高的SPC-A-1细胞株为实验对象，选择0.1μg/mL的鞭毛蛋白，分别用0μg/mL、0.01μg/mL、0.1μg/mL、1μg/mL、10μg/mL的TLR5抗体抑制通路活化，检测细胞内NF-κB荧光素酶的活性，验证TLR5活化通路。构建TLR5-shRNA，转染SPC-A-1细胞48h后，以0.1μg/mL浓度鞭毛蛋白分别刺激SPC-A-1细胞及转染的SPC-A-1细胞，在刺激0min、10min、30min、60min，用Westernblot方法比较TLR5信号通路因子p-IKBα、p-ERK1/2、p-JNK、IKBα、ERK1/2的变化。结果TLR5在肺腺癌细胞株SPC-A-1中呈高表达，且主要表达在细胞膜上。三种细胞株中SPC-A-1细胞NF-κB荧光素酶的活性最高，呈浓度依赖性，0.1μg/mL鞭毛蛋白即可明显增强NF-κB荧光素酶的活性（P0.05）。以适合浓度鞭毛蛋白刺激转染的SPC-A-1细胞，p-IKBα、p-JNK蛋白均未检出，IKBα、ERK1/2蛋白的水平无明显变化（P>0.05）， p-ERK1/2蛋白水平随着时间延长明显增高（P<0.05）。结论外源性配体鞭毛蛋白可激活NSCLC细胞株TLR5蛋白，启动下游信号通路，可能与NSCLC的发生发展有关。

Background and objective It has been proven that toll-like receptor 5 (TLR5) plaied an important role in the development of tumor. In our previous study, we found that the expression of TLR5 was remarkably higher in non-small cell lung cancer (NSCLC) tissues than that in normal tissues, but the activation of TLR5 signaling pathway in NSCLC was still unknown. Te aim of this study is to investigate the expression of TLR5 in diferent types of NSCLC cell lines, and analyze the activity of the signaling pathway afer stimulated by its specific exogenous ligand fiagellin. Methods Te TLR5 protein was detected by immunofiuorescence and Western blot in three kinds of NSCLC cell lines, and the TLR5 mRNA was detected by RT-PCR. Select the cell line of TLR5 highest expression as the research object, and select the suitable concentration of fiagel-lin. NF-κB luciferase activity was detected to validate the TLR5 activation pathway through inhibitory signaling pathways by 0 μg/mL, 0.01 μg

原虫是寄生于鸟的一类常见寄生虫，不仅危害鸟的生长发育，还对人类健康存在潜在威胁。为了解我国鸟类原虫种类与感染状况，作者查阅了近几十年我国在鸟类原虫调查方面的文献。经归类整理，在我国鸟类中共发现原虫70种，隶属于3门、4纲、5目、9科、12属。其中，顶器复合门（Apicomplexa）、孢子虫纲（Sporozoa）、真球虫目（Eucoccidiorida）的原虫有26种，包括艾美耳科（Eimeiridae）的艾美耳球虫属（Eimeria）20种和等孢球虫属（Isospora）2种、隐孢子虫科（Cryptosporidiidae）的隐孢子虫属（Cryptosporidium）3种、住肉孢子虫科（Sarcocystidae）的弓形虫属（Toxoplasma）1种；孢子虫纲、血孢子虫目（Haemospororida）的原虫39种，包括疟原虫科（Plasmodiidae）的疟原虫属（Plasmodium）23种和血变虫属（Haemoproteus）1种、住白细胞虫科（Leucocystozoidae）的住白细胞虫属（Leucocytozoon）15种；肉足鞭毛门（Sarcomastigophora）、动物鞭毛虫纲（Zoomastigophora）、毛滴虫目（Trichomonadorida）的原虫2种，包括单尾滴虫科（Monocercomonadidae）的组织滴虫属（Histomonas）1种、毛滴虫科（Trichomonadidae）的毛滴虫属（Trichomonas）1种；肉足鞭毛门、叶足

Protozoa are a group of common parasites in birds and puts a threaten to public health in humans. In order to keep abreast of the recent protozoan species and infection status in birds in China, we have reviewed a lot of literatures about protozoan studies. In summary, 70 species of protozoa have been recorded in birds of China. These species are classified into 3 phyla (Apicomplexa, Sarcomastigophora, Sarcomastigophora), 4 classes (Sporozoa, Zoomastigophora, Lobosasida, Blastocystidea), 5 orders (Eucoccidiorida,Haemospororida, Trichomonadorida, Amoebida, Blastocystida), 9 families (Eimeiridae, Cryptosporidiidae, Sarcocystidae, Plasmodiidae, Leucocystozoidae, Monocercomonadidae, Trichomonadidae, Entamoebidae, Blastocystidae), and 12 genera. There are 23 species in Plasmodium, 20 species in Eimeria, 15 species in Leucocytozoon, 3 species in Cryptosporidium, 2 species in Isospora, and 1 species in each of Toxoplasm, Haemoproteus, Histomonas, Trichomonas, Entamoeba, Endolimax and Blastocy