牛IFN-γ单克隆抗体的制备及鉴定

为进一步研究牛IFN-γ(BoIFN-γ)单克隆抗体在牛免疫学以及牛疫病诊断中的应用,用原核表达的重组牛IFN-γ(rHis-BoIFN-γ)免疫6周龄雌性BALB/c小鼠,免疫3次后取其脾细胞和骨髓瘤细胞SP2/0进行融合。用建立的间接ELISA方法筛选阳性杂交瘤细胞,经3～5次亚克隆,最终获得12株能稳定分泌抗体的阳性杂交瘤细胞株。Ig亚类鉴定结果显示这12株杂交瘤细胞均为IgG,其中5株为IgG1,3株为IgG2a,4株为IgG2b。这12株杂交瘤细胞在体外具有稳定分泌抗牛IFN-γ单克隆抗体的能力。Western-blot及间接ELISA证明这12株单克隆抗体均可特异性地结合rBoIFN-γ蛋白。间接免疫荧光试验显示,这12株单抗均与真核细胞BHK-21表达的rBoIFN-γ产生荧光反应,证明这些单抗能够与接近天然的BoIFN-γ反应,证实了单抗的特异性。ELISA叠加试验表明,3C2与1H4、1G6与1G4针对不同表位,1A10、1B2、2B9、3C4四株和1C1与1G6、1G4与3C2、1A10与1G8针对相同或相近的表位。     

猪肺炎霉形体热休克蛋白Hsp70单克隆抗体的制备及鉴定

以肺炎霉体体(Mhp)Yin-1株为模板,利用所合成的引物扩增了伴侣蛋白DnaK(热休克蛋白Hsp70)C末端基因,将得到的目的片段分别克隆至表达载体pET-28a和pGEX-6P-1中,原核表达获得了重组蛋白6P-1-DnakC和28a-DnakC,经Western-blot分析,均能被Mhp抗血清识别.以28a-DnakC作为包被抗原,建立了间接ELISA检测方法,以6P-1-DnakC为免疫抗原,免疫BALB/c小鼠.筛选和鉴定了2株分泌抗DnaK的杂交瘤细胞株,命名为1AD10、2AF11.Western-blotting结果表明,这2株单抗与Mhp发生特异性反应而不与其他相关霉形体发生反应.抗体亚类鉴定结果显示,1AD10、2AF111分别为IgG2b和IgG2a亚类,且轻链均为κ链.相加ELISA试验表明,2株单抗能识别不同的抗原表位.     

鹅细小病毒VP1蛋白单克隆抗体的制备及其对应抗原表位区的分析

为获得抗鹅细小病毒(GPV)VP1蛋白的单克隆抗体,以纯化的VP1蛋白为免疫原,免疫4～6周龄的雌性BALB/c鼠,经4次免疫后,取其脾细胞与SP2/0骨髓瘤细胞进行融合,共获得5株能稳定分泌特异性抗体的阳性细胞株。Western-blot结果表明,这5株杂交瘤细胞分泌的抗体均能特异地与纯化后的GPV-VP1重组蛋白发生反应,单克隆抗体亚型鉴定结果表明,3株单抗为IgG1亚型,1株单抗为IgG2b亚型,1株单抗为IgA亚型,轻链均为κ链。用单克隆抗体对分段表达的VP1蛋白进行免疫印迹分析,结果鉴定出3个抗原表位识别区,分别位于VP1的第92～123、92～145和111～145aa。证实,这5株杂交瘤细胞在体外长期培养能稳定地分泌抗GPVVP1的单克隆抗体。     

抗鹅细小病毒NS1蛋白单克隆抗体的制备及对应抗原表位区的分析

以鹅细小病毒非结构蛋白NS1(GPV-NS1)的重组质粒pcDNA3.1-GPV-NS1及纯化的重组蛋白GPV-NS1为抗原,分组免疫4～6周龄的BALB/c鼠,免疫3次后,取其脾细胞与SP2/0骨髓瘤细胞融合,利用间接ELISA方法筛选阳性杂交瘤细胞株,并研究其部分生物学特性.结果共获得6株能稳定分泌特异性抗体的阳性杂交瘤细胞株.单克隆抗体亚型鉴定结果显示,2株为IgG2a型,4株为IgM型,轻链均为κ链.用制备的单克隆抗体对分段表达的NS1蛋白进行免疫印迹分析,结果鉴定出3个抗原表位区,分别位于NS1蛋白的第453～514 aa、485～542 aa、533～598 aa区段.证实,6株杂交瘤细胞在体外长期培养能稳定地分泌抗GPV-NS1单克隆抗体.     

抗鸭坦布苏病毒E蛋白单克隆抗体的制备及鉴定

为了制备抗鸭坦布苏病毒(DTMUV)囊膜(E)蛋白的单克隆抗体,将鸭坦布苏病毒E基因进行原核表达,目的蛋白被纯化后免疫6周龄雌性BALB/c小鼠,按常规的单克隆抗体的制备技术,取其脾细胞与骨髓瘤细胞SP2/0融合,经间接ELISA方法筛选及3次连续克隆化共获得6株能稳定分泌抗E蛋白单克隆抗体的杂交瘤细胞,它们分别被命名为1F3、1G2、1B11、2A5、3B6和4F9。Dot-ELISA及Western-blot分析结果表明,这6株单克隆抗体均能与原核表达的E蛋白特异性结合。经间接免疫荧光(IFA)试验验证,这6株单克隆抗体均能识别鸭坦布苏病毒感染的细胞,且荧光主要位于细胞质中。结果表明,本研究成功制备了6株抗鸭坦布苏病毒E蛋白的单克隆抗体。鸭坦布苏病毒E蛋白单克隆抗体的制备为今后鸭坦布苏病毒快速的病原学诊断以及E蛋白的结构和功能研究奠定了物质基础。     

蓝舌病病毒单克隆抗体的制备及生物学特性鉴定

以纯化的蓝舌病病毒(BTV)11型VP7免疫BALB/c小鼠,运用淋巴细胞杂交瘤技术,借助间接ELISA筛选,获得了3株分泌抗BTV11 VP7的群特异性单克隆抗体(McAb)的杂交瘤细胞株,命名为C12D9、B4 F3、E1 A7.这3株杂交瘤细胞株连续传代3个月再经液氮冻存6个月后复苏培养,仍能稳定分泌特异性McAb.3株McAb均具有ELISA特性和免疫沉淀特性,其中C12D9株上清液的ELISA效价为1256,腹水的ELISA效价为10-6.亚类鉴定,C12D9株为小鼠IgG1,B4F3、E1 A7为IgG2a和IgG2b.琼脂免疫扩散试验和间接ELISA试验表明,这3株均与BTV11型VP7发生特异性反应,为BTV群特异性McAb.     

伪狂犬病病毒gB和gE蛋白单克隆抗体的制备与鉴定

采用大肠杆菌表达系统表达获得了伪狂犬病病毒(PRV)超强毒株ZJ01的gB和gE重组蛋白,用其免疫BALB/c小鼠,通过细胞融合、克隆和间接ELISA筛选,获得了4株能稳定分泌抗PRV单克隆抗体的杂交瘤细胞,细胞培养上清的ELISA抗体效价在1∶1 600~1∶6 400之间,腹水抗体效价达1∶4×105;连续培养20代,抗体效价基本一致。抗gB单克隆抗体B1B6和B3D7属于IgG2b,抗gE单克隆抗体E1B11和E5C10属于IgG1,轻链均为κ型。Western-blot和IFA结果显示,4株单克隆抗体均能与PRV流行毒株ZJ01、HZ、SD、NJ、LA发生特异性反应,B1B6和B3D7还能与PRV gE基因缺失疫苗毒株Bartha-K61发生反应,但E1B11和E5C10不能与Bartha-K61发生反应。     

分泌抗弓形虫单克隆抗体杂交瘤细胞株的建立——体外免疫法制备免疫脾细胞

作者用纯化弓形虫速殖子可溶性抗原体外免疫BALB/C小鼠的正常脾细胞与小鼠SP2/0骨髓瘤细胞融合,经ABC—ELISA和IHA筛选和3次克隆化培养后获得了1株分泌抗弓形虫单克隆抗体的杂交瘤细胞(E5)。E5杂交瘤细胞分泌的单克隆抗体经琼脂双扩散免疫试验确定为IgGl亚类抗体。单克隆抗体的抗原吸收试验结果证明E5单克隆抗体为抗弓形虫特异性抗体。E5杂交瘤细胞的染色体数目为92～108,经连续30代传递培养和4个月冻存复苏试验证明其分泌特异性单克隆抗体的性能相当稳定。初步证实E5单克隆抗体针对一种弓形虫的共同抗原成份。     

猪链球菌2型人源分离株MRP单克隆抗体的制备与鉴定

为建立一种针对猪链球菌2型(SS2)毒力因子的特异性检测方法,研制了抗SS2溶菌酶释放蛋白(MRP)单克隆抗体(McAb),并对其生物学活性进行了分析.将表达、纯化的SS2菌株MRP免疫BALB/c小鼠,采用淋巴细胞杂交瘤技术将免疫鼠脾细胞与SP2/0骨髓瘤细胞融合,以纯化的MRP蛋白作为包被抗原,通过间接ELISA方法筛选获得了6株能稳定分泌抗MRP McAb的杂交瘤细胞系288、3G5、1G1、3G9、186和2C3.经用间接ELISA方法测定,单抗腹水的抗体效价均超过1105,杂交瘤细胞培养上清液的效价为1128～11 024.western-blotting鉴定结果表明,6株单抗均具有特异的生物学活性;以2B8单抗腹水和SS2多克隆抗血清建立的夹心ELISA方法可特异地检测MRP.     

分泌抗布氏锥虫群共同抗原单克隆抗体杂交瘤细胞株的建立

用DEAE-纤维素分离的布氏锥虫伊氏亚种JG克隆株虫体按Segura 氏法制备抗原,常规免疫BALB/c鼠、融合、筛选和克隆。筛选时除使用免疫抗原外,还使用按同法制备的我国不同地区分离的3株布氏锥虫伊氏亚种(JX、ZJ、GY株),1个布氏锥虫马媾疫亚种虫株和培养的1株布氏锥虫指名亚种发育前期型虫体的抗原,以及泰氏锥虫和弓形虫抗原。共获得8个杂交瘤细胞株(4A_(11)、2D_6、5C_8、2E_3、1H_8、1C_9、1F_(11)、4F_6。它们分泌的单克隆抗体与泰氏锥虫、弓形虫抗原均无反应。4A_(11)、2D_6在ELISA和间接免疫荧光试验中只与免疫克隆株抗原反应。其余6株在ELISA中不仅与免疫株抗原反应,而且与其他锥虫虫株抗原呈明显交叉反应,免疫荧光试验则全部阴性。在ID试验中,1C_9、1F_(11)腹水与上述6种抗原呈现明显沉淀线,另6株则均无反应。4A_(11)和2D_6是针对伊氏锥虫群JG克隆株特异抗原,而5C_8、2E_3、1H_8、1C_9、1F_(11)、4F_6则是抗布氏锥虫群共同抗原。     

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.     

U.S. and East Asian Integration

the U.S. is directly impacting on the future of the East Asian community. Therefore, finding ways to get along with the U.S. is crucial to the speed, direction, configuration and character of the East Asian community. In this paper, the author has analyzed the interests of the United States in East Asia and its stands towards the East Asian integration. The author concludes that it is to the interest of the United States to make more efforts to further join in the East Asian integration. And East Asia should accept and welcome the American participation.     

CAFIU Honorary Vice-President Wang Zhizhen Visited Turkey,Sudan and Attended the Third China-Africa People's Forum

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CAFIU Delegation Visited Sri Lanka and Attended the Third China-Africa People's Forum in Sudan

正At the invitation of Sri Lanka-China Friendship Association(SLCFA)and Sudan Network for National Non-Governmental Organizations,a 5-member delegation led by CAFIU Deputy Secretary-General Liu Kaiyang visited Sri Lanka and attended the Third China-Africa People’s Forum held in Sudan from May 7 to 14,2014.As a member of this delegation,I had the great honor     

South Asian NGO Delegation Visiting China

正At the invitation of the Chinese Association for International Understanding(CAFIU),a 10-member NGO delegation composed of representatives from media,think tanks and NGOs from Bangladesh,India and Sri Lanka visited China from April 9 to 18.On the second day of arrival,the delegation visited Xicheng District Administrative Service Center.The center provides such services as     

Building A Platform for Understanding and Establishing Channels for Cooperation——Notes on the 2014 “Understanding & Cooperation” Dialogue

正From June 16 to 18,the 2014"UnderstandingCooperation"Dialogue and Forum on Rethinking Asia,co-hosted by CAFIU and FES and jointly organized with China Foundation for Peace and Development,was held in Tutzing,a beautiful and spacious manor located at 30km southwest of Munich.Nearly 60 delegates from more than 10Asian and European countries participated in the event,including China,Germany,Korea,Japan,     

The Second Sino-US Maritime Security Workshop Held in CIIS

正On September 25-26,2014,CIIS and Hoover Institution,Stanford University jointly held the Second Sino-US Maritime Security Workshop in Beijing.CIIS President Qu Xing and Abraham Sofall,senior research fellow with the Hoover Institution,codelivered the opening remarks.Representatives from China's Ministry of National Defense,     

俄白哈关税同盟的发展及其影响（英文）

In recent years, the Eurasian Economic Community has developed rapidly. Because of the special relationship between the SCO and the Customs Because Customs Union, the latter will inevitably exert some influence on the development of the former, but they are not fundamentally contradictory. Since the of the establishment of the Customs Union, the trade between China and the Customs Union members grows steadily.