禽多杀性巴氏杆菌C_(48-1)成熟外膜蛋白H重组亚单位疫苗免疫效果的研究

将融合表达禽多杀性巴氏杆菌成熟外膜蛋白H(OmpmH)的重组菌pGEX-ompmH／ BL21大量培养,在最佳诱导条件下诱导表达,表达产物经蛋白酶剪切及亲和层析纯化,得到Ompm H基因的原核表达产物,将其与弗氏完全佐剂混合制成油乳剂亚单位疫苗,用该疫苗肌肉注射接种5周龄鸡,首免后每周采血检测抗体,二免后第2周用10 LD50禽多杀性巴氏杆菌强毒菌株C48-1 进行攻击。结果显示,OmpmH具有良好的免疫原性,能诱导鸡体产生特异性抗体,可抵抗强毒菌株C48-1的致死性攻击,免疫效果优于禽多杀性巴氏杆菌弱毒疫苗。     

鸡IL-18对禽多杀性巴氏杆菌DNA疫苗的免疫佐剂作用

为了探索鸡IL-18在禽多杀性巴氏杆菌H基因DNA疫苗中的免疫佐剂作用,分别用共表达鸡IL-18基因和禽多杀性巴氏杆菌C48-1H基因的DNA疫苗、鸡IL-18真核表达质粒pcDNA3/cIL-18与禽多杀性巴氏杆菌C48-1H基因的DNA疫苗混合物、禽多杀性巴氏杆菌C48-1H基因的DNA疫苗肌肉注射5周龄鸡,首免后每周采取外周血及外周抗凝血,应用ELISA和MTT法分别检测免疫鸡的体液免疫及细胞免疫水平。二免后第2周用10 LD50禽多杀性巴氏杆菌强毒菌株C48-1进行攻击。结果鸡IL-18能够明显增强禽多杀性巴氏杆菌H基因DNA疫苗的免疫原性,显著提高免疫鸡的体液免疫和细胞免疫水平,并且鸡IL-18与禽多杀性巴氏杆菌H基因共表达时的免疫佐剂作用最强,能强有力地抵抗强毒菌株C48-1的致死性攻击。结果表明,鸡IL-18可作为DNA疫苗的一种理想的免疫佐剂。     

禽多杀性巴氏杆菌外膜蛋白单价DNA疫苗及融合DNA疫苗的免疫效果分析

为探讨禽多杀性巴氏杆菌omph和ompa基因单价DNA疫苗及融合DNA疫苗的免疫效果,利用PCR技术扩增了禽多杀性巴氏杆菌omph和ompa基因片段,并克隆入pcDNA3.1(+),构建了DNA疫苗pcDNA-OMPH(pOMPH)、pcDNA-OMPA(pOMPA)和pcDNA-OMPH/OMPA(pOMPHA),将其体外转染SP2/0细胞,用间接免疫荧光试验检测其表达情况。同时以弱毒活疫苗作为阳性对照,pcDNA3.1(+)和PBS作为阴性对照,分别免疫4周龄鸡,检测血清特异性抗体水平、外周血淋巴细胞增殖情况和IFN-γ分泌情况,经强毒攻击后计算各疫苗的保护率。结果显示,融合DNA疫苗组血清抗体水平与弱毒活疫苗相当,明显高于单价DNA疫苗(P<0.05);pOMPHA组的刺激值(SI值)和IFN-γ分泌水平显著高于其他各组(P<0.05),单价DNA疫苗组略高于弱毒活疫苗组;强毒攻击后各DNA疫苗均可为动物提供一定的保护力,其中融合DNA疫苗的保护率最高,可达86.7%,其次为弱毒活疫苗,为73.3%,而单价DNA疫苗则低于弱毒活疫苗,仅为60%和66.7%。结果表明,DNA疫苗尤其是融合DNA疫苗在预防禽多杀性巴氏杆菌病(禽霍乱)方面具有较好的应用前景。     

间接血凝试验检测鸡血清中多杀性巴氏杆菌抗体

近几年来,国内外对禽多杀性巴氏杆菌(Pas-teurella multocida)的免疫应答研究表明,体液免疫和细胞免疫在抵抗巴氏杆菌的再感染均有重要的作用。并应用试管凝集反应、间接血凝(IHA)、酶联免疫吸附试验(ELISA)等检测多杀性巴氏杆菌免疫后鸡和火鸡的血清抗体,并认为此3种方法所检出的抗体滴度与对抗强毒的保护作用一致。本研究优化了检测鸡血清抗体的IHA试验及抗原制备的条件,并比较了IHA滴度与攻毒保护的关系。(一)材料和方法1.菌种:中国兽医药品监察所保藏的C_(48)-1强毒菌株,用前通过小白鼠3代,鸡1代复壮,然后接种于血液马丁肉汤或血液马丁琼脂培养,培养移植不超过3次,菌液10~(-8)稀释0.1ml(含2～     

禽多杀性巴氏杆菌交叉保护因子的研究

禽多杀性巴氏杆菌交叉保护因子存在活鸡体培养物中。本文用鸡 活体培养禽多杀性巴氏杆菌C_(48-1)强毒株(血清1型),在菌血症时采血经 差速离心收获细菌,制成的死菌苗对异型菌株C_(44-42)(血清3型)和P_(1059) (血清3型)攻毒产生交叉保护作用。而41℃肉汤培养的C_(48-1)菌株则不具有这种交叉保护作用。 活体培养的细菌经冻融、溶菌酶、透明质酸酶、DN_(ase)、EDTA、Triton X-100等处理溶解后,细菌溶解物对异型菌株保护率达100％,而其溶解物的上清和沉淀时异型菌株保护率也达66.7％及80％。而溶解物沉淀经蛋白酶处理后,则失去了交叉保护能力,说明产生交叉保护作用的交叉保护因子(CPF)可能是蛋白质。 生化分析表明,活体培养的禽多杀性巴氏杆菌和肉汤培养的禽多杀性巴氏杆菌其物质含量是不同的。活体培养的细菌溶解物含CPF,其蛋白与糖含量比(P/C)为3.97:1,而肉汤培养的细菌P/C为1.01:1。SDS-PAGE分析表明,有一分子量在17200～30000之间的蛋白成分仅存在于活体培养的细菌中,经测算其分子量大约为19320。这些研究表明,分子量为19320的蛋白质可能就是存在于活体培养物中的交叉保护因子。     

鸭源大肠杆菌与多杀性巴氏杆菌融合菌株DB4免疫原性的研究

本研究旨在评价融合菌株DB4的免疫原性以及对亲本大肠杆菌ZYD2(O78)株、亲本多杀性巴氏杆菌YB2(5∶A)株的攻毒保护力。制作DB4蜂胶灭活疫苗分为一免组、二免组对鸭进行免疫,对两组鸭的血清抗体效价、IL-4和IFN-γ水平、外周血淋巴细胞增殖活性、白细胞吞噬活性进行测定,并于首免后2周以大肠杆菌ZYD2(O78)、多杀性巴氏杆菌YB2(5∶A)攻毒。结果显示,DB4能够诱导鸭产生两种亲本抗体,效价达1∶29、1∶28,同时能刺激淋巴细胞增殖活性和白细胞吞噬活性,免疫后刺激指数增加3倍,与对照组差异显著(P0.05),白细胞吞噬率升高30%,血清中IL-4和IFN-γ含量分别高于对照组33.5、12.5 ng/L,可抵抗亲本菌株的攻击,保护率80%,表明DB4具有良好的免疫原性,可应用于研制大肠杆菌和多杀性巴氏杆菌二联疫苗。     

牦牛源多杀性巴氏杆菌外膜蛋白OmpH和OmpA重组亚单位疫苗对小鼠的免疫效果

为分析牦牛源多杀性巴氏杆菌(Pasteurella multocida)外膜蛋白H(OmpH)和外膜蛋白A(Om-pA)的免疫效果,构建了牦牛源多杀性巴氏杆菌OmpH和OmpA的原核表达载体pET-28a-OmpH和pET-28a-OmpA,表达并纯化重组外膜蛋白H(rOmpH)和重组外膜蛋白A(rOmpA),制备相应的单价疫苗和二价疫苗(rOmpH-A)以及全菌灭活疫苗,以PBS作为对照,皮下免疫小鼠。首免后每周采血1次,用间接ELISA检测抗体动态变化,并于三免后第9天用2LD50的分离菌株进行攻毒,比较保护力。结果,各试验组均能产生特异性抗体,全菌灭活组和rOmpA组小鼠的存活率分别为80%、40%,而rOmpH组、rOmpH-A组小鼠全部死亡,rOmpH和rOmpH-A组仅推迟小鼠死亡的时间,说明保护力微弱,不能抵抗同源菌株的致死性攻击。结果表明,rOmpA蛋白具有保护力,而rOmpH不具有保护力。     

多杀性巴氏杆菌prfA基因的序列分析与原核表达

为探索多杀性巴氏杆菌的致病机理,采用PCR方法分别扩增了家兔源和禽源多杀性巴氏杆菌C51-17、Pm898、Pm8916、Pm901、Pm9616、PmZM、PmQin菌株的prfA基因,并对其进行序列分析。将C51-17株的prfA基因克隆到质粒pET30a中,将重组表达载体pET30a-prfA转化到大肠杆菌BL21(DE3)pLysS感受态细胞中,经IPTG诱导表达,进行SDS-PAGE分析和Western-blot分析。结果显示,这7株菌株与GenBank登录的3株多杀性巴氏杆菌之间prfA基因在核苷酸水平上的同源性为95.6%~100%,在氨基酸水平上的同源性为98.9%~100%。诱导表达的重组蛋白PrfA的分子质量约为45.6ku,与预期的大小一致,以可溶性形式表达。重组蛋白PrfA可与家兔抗多杀性巴氏杆菌阳性血清反应,具有良好的反应原性。     

禽巴氏杆菌免疫鸡IgG、IgM消长规律的研究

本试验利用鸡血清中主要有IgG、IgM(IgA主要在粘膜),和2-基巯乙醇可破坏IgM的理论,以间接血凝法检测禽霍乱731、GNCoⅡ弱毒苗免疫鸡和C_(48-1)强毒接种后发病治愈鸡(即模拟禽霍乱强毒感染鸡)的IgG、IgM消长规律。结果抗体中主要有IgM;当强毒攻击后,三个组均无回忆反应的迹象,其抗体中仍是IgM占优势。对731、GNCo Ⅱ和C(48-1)的荚膜粗提物中多糖与蛋白成分的分析表明,多糖是蛋白的15.35倍以上。荚膜在菌体外层,与机体紧密接触,又为TI抗原,易刺激机体产生IgM,因此产生IgM占优势是巴氏杆菌本身结构块定的。然而如将荚膜抗原与某蛋白载体结合后转变成TD抗原,即可诱导机体产生IgG和回忆反应,这样免疫期可望能延长。目前研制的多种禽霍乱弱毒苗免疫期较短,本试验人工模拟强毒感染康复鸡的免疫期亦较短,从试验结果看出,由于禽霍乱菌免疫主要产生IgM,很少产生IgG,而IgM比IgG消失快的特点及无回忆反应的特点,初步认为是禽霍乱菌免疫期短的原因。     

牛A型多杀性巴氏杆菌三聚体自转运蛋白免疫保护性的研究

为研究多杀性巴氏杆菌三聚体自转运蛋白(TAAs)的免疫保护效果,在对牛A型多杀性巴氏杆菌Pm CQ2株假定TAAs编码基因Pm1g0001的黏附基序分析基础上,对其黏附基序进行PCR扩增,并与质粒p ET-32a(+)连接,将重组质粒转入BL21(DE3)诱导表达获得重组蛋白,纯化后与佐剂混合免疫小鼠,三免后收集小鼠血清检测抗体效价,并用Pm CQ2菌液进行攻毒保护性试验。结果显示,Pm1g0001的黏附基序具有TAAs的典型结构,重组蛋白r Pm1g0001免疫小鼠后可以刺激机体产生高效价抗体,对Pm CQ2的保护率达60%,具有良好的免疫保护效果,提示牛A型多杀性巴氏杆菌三聚体自转运蛋白可以作为研究新型亚单位疫苗的候选靶标。     

The Development of EU-Russian Relations

The EU＇s strategy for Russia since the end of the cold war has to a large extent failed, and the direct consequences are the currently deadlocked EU-Russia relationship. There are three reasons for this： the EU and Russia＇s respective strength and status relative to each other have been changing since the beginning of this century; the EU＇s strategy towards Russia has been more fragmenting in recently years; and the American factor. The EU-Russia relations could continue to be in poor shape in the years to come, and it is very likely sometimes even further worsening. But the two sides would refrain from extreme measures and prevent their relations from sliding into real cold war.     

Young American China Watchers＇ Views on China

In this paper, the author has made a thorough study over the new generation of American China Watchers. Most of the pages are devoted to a detailed comparison between the old generation and the young generation of American China Watchers regarding to their background, education, and views over China. At the end of the paper, the author also gives some policy suggestions for both the Chinese and U.S. government based on the findings in this study.     

The Balance Between Coercion and Accommodation

1. Introduction
On the morning of April 1st, 2001, a U.S. EP-3 surveillance aircraft was conducting an intelligence flight along the Chinese territorial border in the airspace south-east of China＇s Hainan Island. Two Chinese fighter jets intercepted the spy plane and followed its flight, as is considered ordinary international practice. For reasons unknown, the EP-3 airplane made a sudden maneuver and crashed into one of the Chinese planes. The Chinese plane dropped into the sea, the pilot missing. The U.S. spy plane was also severely damaged. It intruded into Chinese territorial space and landed on Hainan Island without official permission from the Chinese side. After landing, the spy plane and 24 crew on board were detained by the Chinese authorities. This air collision episode was the first of its kind in the China-U.S. relations and an urgent diplomatic crisis ensued.     

中国需要构建怎样的周边(英文)

正The new central leadership has highlighted the strategic importance of better relations with neighboring countries and the construction of a solid foundation for China’s neighborhood strategy so as to create favorable conditions for Chinese development.1China is strengthening top-level strategy in foreign relations,focusing on global-level relations and staying committed to relations     

中巴关系的发展及其认识误区（英文）

China and Brazil established a strategic partnership in 1993. The bilateral ties have seen rapid development in almost all areas. However, there are eight myths surrounding the China-Brazil relations. These myths are the result of a lack of mutual understanding as well as the misconceptions result misconceptions of the western observers and media. These misunderstandings have of have jeopardized images of the China-Brazil relations and also hindered further development of their bilateral relationship.     

中国的“主场外交”:机遇、挑战与任务（英文）

The successful hosting of the CICA summit has drawn great attention The attention to China's "host diplomacy". Analyzing new features of China's "host to "host diplomacy" during this new period, this paper will look at the opportunities and challenges facing China's "host diplomacy" and discuss the problems that China should address in order to do a better job in this regard and bring its "host diplomatic advantages" into full play.     

亚洲整体性崛起及其效应(英文)

Since the beginning of the 21st century, the overall rise of the Asian geographic plate has been particularly prominently. The overall rise of Asia reflects the improvement of a large number of developing countries in Asia. The long-occupied aggressive status of the United States and Europe is weakening, and the overall rise of Asia appears more highlighted, and its impact upon the world seems more intense.     

CAFIU Convenes the 11~(th) Council Meeting in Beijing

正On the morning of April 2,2014,CAFIU held the 11th Executive Council Meeting in Beijing.Present at the Meeting were Member of the Politburo of the 17th CPC Central Committee and Vice-Chairman of the 11th CPPCC National Committee Mr.Wang Gang,ViceChairman of the 11th NPC Standing Committee and President of CAFIU Mr.Zhou Tienong,ViceChairpersons of the NPC Standing Committee Mme     

Speech by Honorary President Zhou Tienong at the 11~(th) Executive Council Meeting of CAFIU (Excerpt)

正I am very pleased to attend the 11th Executive Council Meeting of CAFIU,and would like to express my hearty congratulations to President Yan Junqi,the new leadership and the newly-elected Executive Council.I also want to express my sincere thanks for choosing me to be the Honorary President of CAFIU!     

Name-list of Members of the Standing Committee of the (11)~th Executive Council (in alphabetical order of the surname's first letter)

正I.Political Parties and Government Sector Lu Yan Director,Beijing Municipal Commission of Commerce Tong Guili Member,Standing Committee of the CPC Municipal Committee of Hangzhou,Zhejiang Province Secretary,CPC Working Committee,Administrative Commission for the Sci-Tech and Innovation Industrial Park in west Hangzhou City