乳牛L-选择素单克隆抗体的制备与亚类鉴定

以乳牛L-选择素为免疫抗原免疫BALB/c小鼠1只,相隔14 d进行3次免疫,抗原用量每次30μg,首次免疫后第116 d进行尾静脉加强免疫,抗原用量10μg。免疫脾细胞与骨髓瘤细胞融合后接种12块96孔板。以L-选择素包被酶标板,间接ELISA检测阳性孔,有限稀释法克隆阳性孔,筛选出6株杂交瘤细胞。小鼠腹腔注射杂交瘤细胞株制备McAb,并对其分别进行Ig亚类测定。结果,6株杂交瘤细胞7B10、10F6、10E2、5C6、12G7和11A3的Ig亚类分别是IgG2a、IgG2aI、gG2bI、gG1I、gG1和IgG1。     

番鸭呼肠孤病毒σB蛋白单克隆抗体的制备及鉴定

以纯化复性的原核表达的番鸭呼肠孤病毒(MDRV)σB蛋白为抗原免疫BALB/c雌性小鼠,按常规方法制备单克隆抗体,通过间接ELISA、Western-blot和免疫组织化学试验进行筛选及鉴定.结果显示,获得4株能够稳定分泌抗σB蛋白单克隆抗体的杂交瘤细胞株(A3F、B2B、C3E和D2G),其亚型分别属于IgG2b、IgG2b、IgM和IgM.采用小鼠体内诱生法制备的腹水均能与MDRV的σB蛋白发生特异性反应,表明,制备的单克隆抗体能够识别天然构象的σB蛋白.     

羊口疮病毒B2L截短重组蛋白刺激小鼠产生特异性抗体的研究

为研究羊口疮病毒(ORFV)新型亚单位疫苗,设计了ORFV B2L优势抗原表位截短重组蛋白,命名为HBBH并经原核表达和鉴定。以不同剂量HBBH不加或加不同佐剂,经滴鼻或颈部皮下注射免疫BALB/c小鼠,其中滴鼻共4组(HBBH 10μg、20μg、30μg和20μg+LTB),注射组(HBBH 20μg+201佐剂)。同时设ORFV组织灭活抗原+201佐剂注射免疫对照和空白对照,每组共免疫3次,每次间隔7 d。HBBH间接ELISA检测一免、二免、三免后7 d以及三免后14 d血清中ORFV IgG、IgA和三免后14 d肺灌洗液sIgA的抗体水平,取不同水平的IgG ELISA阳性血清检测细胞中和抗体效价。结果显示,表达并鉴定了具有良好反应原性的包含ORFV B2L蛋白优势抗原表位的截短重组蛋白HBBH。免疫小鼠试验结果表明,不同剂量HBBH通过滴鼻或颈部皮下注射均可刺激小鼠产生ORFV特异性IgG抗体,20μg HBBH+201佐剂注射免疫能最大刺激小鼠血清IgG抗体的产生。实验小鼠三免后14 d肺灌洗液特异性sIgA仅在HBBH滴鼻免疫剂量≥20μg的三个组中检测到,其中30μg HBBH组刺激小鼠产生了最高水平的sIgA。在同为20μg HBBH滴鼻免疫时,添加LTB佐剂刺激小鼠产生了更高水平的sIgA。表明一定剂量的HBBH+LTB佐剂滴鼻免疫可更好地刺激小鼠产生sIgA。HBBH间接ELISAIgG阳性血清的细胞中和抗体效价可高达1:128,且能中和4株ORFV分离株。结果表明,20μgHBBH+201佐剂注射免疫3次和20μg HBBH+LTB佐剂滴鼻免疫3次均能刺激BALB/c小鼠产生高水平的抗ORFV血清中和抗体IgG和黏膜抗体sIgA。本研究为ORFV B2L优势抗原表位截短重组蛋白作为亚单位疫苗提供了数据支持。     

禽脑脊髓炎病毒单克隆抗体的制备与初步应用

利用提纯的禽脑脊髓炎病毒(AEV)Van Roekel株作为免疫原,免疫6周龄BALB/c小鼠,采取其脾细胞与SP2/0骨髓瘤细胞融合,用间接ELISA法筛选,间接免疫荧光法(IFA)和免疫组织化学法鉴定,经3次亚克隆得到了稳定分泌抗AEV单克隆抗体的杂交瘤细胞株F11和G2,制备了腹水,并利用该单克隆抗体初步建立了Dot-ELISA、间接ELISA和IFA等特异性检测AEV抗原的方法。     

A型流感病毒PB1蛋白单克隆抗体的制备及鉴定

以A型流感病毒A/Chicken/Zhejiang/YH/2/2013(H7N9)的PB1基因为模板,扩增其1519～2274bp间的片段,并构建原核表达载体pET-28a(+)-H7N9-sPB1。通过大肠杆菌原核表达获得重组蛋白His-sPB1后,以其作为抗原免疫BALB/c小鼠,通过细胞融合筛选获得3株能够稳定分泌单克隆抗体的细胞株5G5、2H3和5E8。Western-blot和IFA检测表明,3株单克隆抗体对于不同亚型的流感病毒均具有良好的反应性。免疫共沉淀结果表明,3株单克隆抗体能用于免疫共沉淀试验。A型流感病毒PB1蛋白单克隆抗体的制备为进一步研究PB1蛋白的结构与功能以及流感病毒的监测与诊断提供了必要的工具。     

犬瘟热病毒H蛋白单克隆抗体的制备与鉴定

为制备犬瘟热病毒(CDV)H蛋白单克隆抗体,采用蔗糖密度梯度离心法提纯CDV-Ondetstepoort株,利用RT-PCR方法扩增其H基因截短片段(H1、H2、H3),克隆至pET-28a (+)载体并转化至大肠杆菌Rosetta感受态细胞中进行原核表达,用Ni-NTA亲和层析柱纯化3种截短H蛋白;以纯化病毒为抗原免疫BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,采用间接ELISA、间接免疫荧光法(IFA)和病毒中和试验分析杂交瘤细胞的生物学特性。结果表明,纯化的截短H蛋白表达正确,以此筛选到7株能稳定分泌犬瘟热病毒和重组H2蛋白单克隆抗体的杂交瘤细胞株(3B1、3B5、3C5、3D5、5A6、4G12和7B2),抗体亚类鉴定均为Ig G1κ;IFA检测均为阳性,表明7种单克隆抗体特异性好;病毒中和试验测定其培养上清的抗体中和效价为24～28,诱生BALB/c小鼠腹水的抗体中和效价为1×102～1×106;间接ELISA叠加试验显示,3B5、4G12、5A6、7B2之间的叠加系数均在50%以上,表明这4种单克隆抗体识别不同抗原表位;另外,3B5、4G12、5A6、7B2还能中和当前流行的Asia 1型CDV毒株,具有较好的广谱中和活性。本研究为后续CDV感染的免疫治疗和CDV检测方法的建立提供了材料基础。     

抗O型口蹄疫病毒VP1蛋白单克隆抗体的制备及鉴定

以高浓缩的O型口蹄疫病毒(FMDV)灭活疫苗为抗原免疫BALB/c小鼠,取其脾细胞与小鼠SP2/0骨髓瘤细胞进行融合,以重组表达的O型重组VP1蛋白(O-r VP1)为筛选抗原,利用杂交瘤技术获得1株可稳定分泌抗O型FMDV VP1蛋白抗体的杂交瘤细胞株1C7,染色体数高于任一亲本细胞的染色体数目;腹水的抗体效价为1︰105,质量浓度为4.7 mg/m L。1C7单克隆抗体为IgG2b亚型,相对亲和力常数为1.5 mol/L,能够与O型FMDV特异性地结合,且仅与结构蛋白VP1反应。稳定性鉴定结果表明1C7株能稳定分泌抗体。阻断ELISA试验表明1C7单克隆抗体能够被O型FMDV免疫阳性血清特异性地阻断。本研究为进一步建立O型FMDV抗体的阻断ELISA及胶体金免疫层析快速检测方法奠定了基础。     

牛源金黄色葡萄球菌与无乳链球菌荚膜多糖蛋白结合疫苗的研制与免疫原性研究

为了评价牛源金黄色葡萄球菌J581株和无乳链球菌A20株荚膜多糖蛋白结合疫苗的免疫原性,对制备的3批荚膜多糖进行了多糖、蛋白质及核酸含量检测,同时分别用纯化的J581和A20株荚膜多糖与破伤风类毒素偶联制备了质量浓度分别为5、10、15μg/m L的6种荚膜多糖蛋白结合疫苗及质量浓度各为10μg/m L的J581和A20株荚膜多糖蛋白结合混合疫苗,用这几种疫苗对小鼠进行免疫,采用间接ELISA方法及Ig G试剂盒对免疫前后不同时间段的小鼠血清进行抗J581和A20株荚膜多糖的特异性抗体水平及Ig G抗体效价检测。结果表明,制备的3批纯化荚膜多糖中,J581多糖平均质量浓度为49.78 mg/L,A20多糖平均质量浓度为55.76 g/L,2种多糖中蛋白和核酸含量均低于1%,符合疫苗生产要求;小鼠免疫试验结果表明,6种不同多糖含量的荚膜多糖蛋白结合疫苗和荚膜多糖混合疫苗均能刺激机体产生一定水平的特异性抗体效价及Ig G抗体,且在第28天抗体水平达到最高,其中荚膜多糖蛋白混合疫苗组中J581和A20抗体效价及特异性Ig G水平均显著高于6种单疫苗组（P<0.05）。本研究为研制金黄色...     

猪繁殖与呼吸综合征病毒N蛋白单克隆抗体的制备与鉴定

为了鉴定猪繁殖与呼吸综合征病毒(PRRSV)N蛋白抗原表位,本研究将PRRSV N蛋白进行了原核表达和纯化,并用纯化后的N蛋白免疫BALB/c小鼠。利用淋巴瘤细胞融合技术和间接ELISA抗体筛选方法,制备获得4株稳定分泌单克隆抗体的杂交瘤细胞株3D10、3G12、6E2和6F4,Western-blot和IFA鉴定其均能与PRRSV特异性反应。4株单抗均属于Ig G1亚类,轻链均为κ链。ELISA结果显示,3D10、3G12、6E2和6F4之间的叠加系数均在50%以上。选择N蛋白3个主要抗原表位合成肽进行ELISA检测,证明4株单抗识别的抗原表位均为30～52 aa,提示4株单克隆抗体亲和力可能存在一定差异,为该病毒研究提供了重要材料。     

鸭疫里氏杆菌脂多糖单克隆抗体的制备

将鸭疫里氏杆菌CH3株作为免疫原免疫BALB/c小鼠,细胞融合后将CH3株脂多糖(LPS)作为包被抗原,筛选阳性杂交瘤细胞株。共获得2株稳定分泌鸭疫里氏杆菌CH3株LPS单克隆抗体的杂交瘤细胞株,分别将其命名为7H1和8A9。2株单克隆抗体均为IgG1亚类。腹水单克隆抗体ELISA效价分别为1∶12 800和1∶6 400。玻片凝集试验和悬浮荧光试验检测结果表明,2株单克隆抗体与血清1型菌株WJ4发生特异性反应,而与鸭疫里氏杆菌血清2型菌株NJ-3及血清10型菌株HXb2无反应性。Westernblot结果表明,2株单克隆抗体与LPS的反应条带位于LPS O抗原部分。本研究成功制备了稳定性好、特异性强的LPS单克隆抗体,可用于进一步研究鸭疫里氏杆菌LPS的结构和致病机制。     

China＇s Rise vs. International Order Evolution

A heated debate over the rise of China has been going on unabated in the international community for nearly two decades. However, with the elapse of time, the initial divergence of views in the mainstream opinion has gradually diminished and the gap in China policy among nations has likewise narrowed down. Instead, reason and a realistic approach have eventually taken center stare.     

The U. S. Policy toward DPRK: Opportunities and Constraints

布什政府上台以来,始终未能阻止朝鲜拥核步伐.朝核试后,美对朝政策有了新的调整,六方会谈得以复会.但美朝双方受各自国家利益和政策目标的限制,在朝鲜半岛无核化问题上的尖锐分歧难以解决,朝核危机仍存在再度升级的可能.     

Marxist Philosophy vs. International Relations Research in China

This article argues for the confirmation of Marxist philosophy as the ideology that guides international relations （IR） studies in China. Justification comes through the following points. According to Marxism, correct theoretical guidance serves as the prerequisite for correct practice. Marxist philosophy embodies, by nature, righteousness, in opposition to political hegemony. Accordingly, practice under its guidance represents fruitful revolutionary realism. Finally, advanced studies of international relations in this country cannot be separated from philosophical thinking. Using these arguments as background, this article specifically advocates materialism, patriotism, and service to the people as the corner-stone of China＇s IR theory building.     

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.     

Excavating Concealed Tradeoffs: How Russians Watch the News

Citizens in democracies are expected to make better decisions if they understand policy tradeoffs. However, politicians rarely have incentives to communicate them; citizens are uncomfortable choosing among valued outcomes; and devising a common metric is difficult. It is not surprising that in the United States the environment provides relatively little cuing or priming of tradeoffs in television news. Russian citizens, on the other hand, face a media environment in which tradeoff cuing is intentionally suppressed by owners' agendas, yet viewers detect concealed tradeoffs even in the absence of tradeoff priming and viewpoint diversity. Analysis of discourse among ordinary Russians in 16 focus groups convened in four cities, differentiated by political reform and media market environments, showed that when watching news in which tradeoffs are thoroughly concealed, viewers challenge stories by offering a broad spectrum of uncued tradeoffs. Tradeoffs come from diverse policy domains and represent a range of cognitive strategies, some of which are considerably more abstract than others and link elements of their observations and assumptions (together with what they can extract from the stories) into complex reasoning outcomes.     

The U.S. Global Strategy as Seen from the Debate in the Primary Election

There may be more than one year to go before the 2008 U.S. presidential election, but the primary election of the two parties has already begun ahead of time. Both the Democratic and Republican candidates for the     

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.     

Constitution of the Chinese Associa tion for Interna tiona l Understa nding(Approved by the Plenary Session of the Eleventh Executive Council on April 2, 2014)

正Chapter I General Rules Article 1 The Chinese Association for International Understanding is a national non-governmental organization(NGO)in the People’s Republic of China.Article 2 The official name of the Association is"Zhong Guo Guo Ji Jiao Liu Xie Hui"in Chinese,or"Jiao Liu Xie Hui"for short.Its English translation is"Chinese Association for International Understanding",and abbreviated as"CAFIU".     

Guide to Authors

正Published by China Institute of International Studies,China International Studies is China’s first English-language journal on diplomacy and international politics for formal circulation.The magazine mainly publishes the latest research achievements of Chinese scholars and specialists on China’s diplomacy,international politics and international relations under such columns as China’s Diplomacy,International Relations,International Politics,International Security,Regional Issues and World Economy.