温和气单胞菌单克隆抗体的制备及特性分析

利用细胞融合技术建立了5株分泌抗温和气单胞菌CR79-1-1株单克隆抗体(McAb)的杂交瘤细胞系,通过特异性鉴定获得两株与参考菌株中的温和气单胞菌发生反应,但与嗜水气单胞菌以及鳗弧菌、爱德华氏菌、克鲁氏耶尔森菌、大肠杆菌、荧光假单胞菌均不反应的杂交瘤细胞系,分别命名为2G3和1A4.快速酶联免疫分析法(ELISA)测得2G3和1A4的亲和常数分别为1.72×108和5×108M-1;应用方阵配对实验证实,2株单抗针对特异性抗原上不同表位.     

溴氯海因对鱼类的急性毒性试验及治疗效果

溴氯海因浓度为0.3×10-6,0.6×10-6和1.5×10-6时,4种试验鱼(草鱼(C tenopharyngodon idellus),白鲢(Hypophthalm ichthys m olitrix),日本鳗(Anguilla japonicus),红剑尾鱼(Xiphophorus helleri))是安全的;浓度为2×10-6时,前2种试验鱼全部存活,而日本鳗和红剑尾鱼的死亡率为7.5%和15%.并测定了该药对嗜水气单胞菌嗜水亚种(Aerom onas hydrophila hydrophila)、荧光假单胞菌(Pseudom onas fluorescens)、鲁氏不动杆菌(Acinetobacter lwoffi)及迟钝爱德华氏菌(Edwardsiella tarda)4种菌的最小抑菌浓度,分别为0.312 5μg.mL-1、0.156 0μg.mL-1、0.156 0μg.mL-1、0.312 5μg.mL-1.用浓度为0.6×10-6,0.3×10-6和0.15×10-6的溴氯海因预防和治疗白鲢细菌性出血病和患爱德华氏菌病的日本鳗,均取得一定的防治效果.     

环氧合酶-2单克隆抗体的制备和初步应用

以合成肽COX-2-KLH为完全抗原免疫BALB/c 小鼠,通过脾细胞与骨髓瘤细胞SP2/0融合的杂交瘤技术,筛选获得4株(1E10、3D6、5B11、6G4)稳定分泌抗COX-2单克隆抗体(McAb)的杂交瘤细胞株.采用ELISA的方法鉴定McAb效价、亚型及特异性.各株McAb均属IgG1 亚型,其中1E10腹水纯化后MCAb效价在1∶107以上,SDS-PAGE电泳为单一条带,与同源异构体蛋白不发生交叉反应.获得的McAb被成功应用于COX-2生物学研究中的免疫组织化学(IHC)和Western Blot试验中.本研究获得的单抗具有特异、高效的特点,为COX-2生物学相关研究提供理论依据.     

抗人FXYD3单克隆抗体的制备与初步鉴定

【摘要】目的 制备抗人FXYD3单克隆抗体（McAb）并鉴定其生物学特性。方法 以固相合成法获得的FXYD3合成多肽与载体蛋白KLH偶联后免疫BALB/c小鼠,采用杂交瘤技术制备抗人FXYD3 McAb。用ELISA 、Western Blot、免疫组化技术对抗人FXYD3 McAb的亚型、效价、亲和力及特异性进行鉴定。结果 筛选出两段较好的优势抗原表位,分别为NDLEDKNSPFY和KFGQKSGHHPGETPPLITPGSA获得四株可稳定分泌抗人FXYD3 McAb的杂交瘤细胞株02A12C4、02F1E8、02E6B10与03A10E11,亚型鉴定重链均为IgG1,轻链为kappa链。间接ELISA法测定效价均在为1∶104以上。其中02F1E8效价为1∶105以上,亲和常数为3.11?08 L / mol。免疫组化分析显示了FXYD3均匀分布于肝癌细胞与人胰腺癌Bxpc-3细胞系细胞的胞膜上。结论 成功制备了四株抗人FXYD3 McAb,其中02F1E8纯度好、效价高及特异性强。为进一步研究FXYD3在肿瘤组织及细胞系中的表达及临床意义奠定了基础。     

杀鲑气单胞菌单克隆抗体的制备及其特性分析

应用杂交瘤单克隆抗体技术制备了6株分泌抗杀鲑气单胞菌杀鲑亚种的单抗细胞株,并对其特性进行分析。结果显示：6株单抗中IgM有3株,IgG1有2株,IgG2a有1株,且抗体效价为1：12800-1：51200,检测灵敏度为1．0×10^5-1．0×10^8cfu·mL^-1。进一步实验证实这些单抗与其他病原菌都无交叉反应。但单抗5C7、7H6与杀日本鲑亚种有交叉反应;单抗8A2与无色亚种存在阳性反应。表明杀鲑气单胞菌亚种之间既有独特的抗原决定簇,又有共同抗原位点。制备的单抗可用于杀鲑气单胞菌的快速诊断和亚种鉴定,为该菌的进一步研究提供必要手段。     

基因工程人表皮生长因子（rhEGF）单克隆抗体的制备与初步鉴定

以基因工程人表皮生长因子(rhEGF)为抗原,免疫BALB/c小鼠,将免疫的小鼠脾细胞与小鼠Sp2/0-Ag14骨髓瘤细胞进行融合,经多次筛选和再克隆化,获得3株能稳定分泌抗rhEGF单克隆抗体(McAbs)的杂交瘤细胞株.对该3株杂交瘤细胞小鼠腹水进行了抗体效价测定,结果显示在1×10-6～2×10-7之间.对上述3种杂交瘤单抗(分别命名为:AE-2A4,AE-2G5和AE-3E11)进行了特异性、抗原决定簇及相对亲和力测定.结果表明,它们都具有抗rhEGF高特异性;并分别抗两个不同抗原决定簇;相对亲和力大小次序为:AE-3E11>AE-2A4>AE-2G5.Ig亚类鉴定结果显示:AE-2A4,AE-3E11同为IgG1亚类,而AE-2G5为IgG2b亚类.确定AE-2A4单克隆抗体可用于制备高纯rhEGF.     

鱼类致病性迟钝爱德华氏菌胶体金快速检测试纸的研制

制备迟钝爱德华氏菌胶体金快速检测试纸并对其性能进行检测.以柠檬酸三钠作为还原剂制备胶体金颗粒,标记迟钝爱德华氏菌多克隆抗体,将羊抗兔IgG和纯化后的迟钝爱德华氏菌多克隆抗体包被于NC膜上作为质控线和检测线,以检测样品中的迟钝爱德华氏菌(E.tarda).结果表明:所研制的试纸特异性良好,对大肠杆菌、金黄色葡萄球菌、鳗弧...     

心肌肌钙蛋白I-C复合物酶联免疫检测方法的建立

为了制备抗人心肌肌钙蛋白I(cTnI)单克隆抗体、建立检测人cTnI链酶亲和素-生物素双抗体夹心酶联免疫吸附测定(ELISA)方法,用基因工程表达的人cTnI免疫雌性BALB/c小鼠,以杂交瘤技术制备得到三株特异性抗人cTnI的单克隆抗体记为6G3,6A6,3G9.三株杂交瘤细胞中6G3和6A6为IgG1,3G9为IgG2b,腹水效价分别为4×10-5,4×10-5和10-5.亲和常数为5.0×109,6.4×109和3.8×109mol-1.Westernblotting结果表明6G3和6A6能够识别cTnI-C复合物.把6G3抗体生物素化并制备抗cTnI、cTnC和cTnI-C的大鼠多克隆抗体.以多抗作为固相捕捉抗体,生物素化6G3抗体作为检测抗体,建立检测cTnI的双抗体夹心ELISA方法并初步应用于临床病人标本检测.以cTnI-C复合物多抗捕捉检测方法具有良好的灵敏度、特异性和准确性,灵敏度为0.9ng/mL,较包被其他抗体捕捉方法提高1~2倍.批内变异系数为5.03%,回收率为94.56%.测定17例AMI病人和18例血清样本临床诊断结果符合率为100%.     

人绒毛膜促性腺激素β亚基(hCG-β)单克隆抗体的制备及鉴定

为了获得抗人绒毛膜促性腺激素β亚基(hCG-β)单克隆抗体,以hCG为免疫原,经细胞融合和有限稀释后,得到4株分泌抗hCG-β单克隆抗体(McAb)细胞株.酶联免疫吸附测定(ELISA)结果表明,所得抗体均为IgG1,κ型,其腹水效价均可达10-6.     

毒品苯环利啶(PCP)单克隆抗体的制备及鉴定

将毒品苯环利啶(PCP)与牛血清白蛋白(BSA)偶联形成复合抗原免疫Balb/c小鼠,获得6株稳定分泌抗PCP单克隆抗体(McAb)的杂交瘤细胞株.它们的腹水效价在1∶105～1∶106.抗体类型鉴定结果表明,属于IgG1,κ型.这些抗PCP单抗均可用于毒品的免疫检测.     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

An Analysis of Catherine＇s Character——Wuthering Heights

Wuthering Heights, Emily Bronte＇s only novel, was published in December of 1847 under the pseudonym Ellis Bell. The book did not gain immediate success, but it is now thought one of the finest novels in the English language. Catherine is the key character of this masterpiece, because everybody and everything center on her though she had a short life. We can understand this masterpiece better if we know Catherine well.     

Lower Paleozoic oil relationships within Williston Basin, Canada

The Williston Basin is a significant petroleum province, containing oil production zones that include the Middle Cambrian to Lower Ordovician, Upper Ordovician, Middle Devonian, Upper Devonian and Mississippian and within the Jurassic and Cretaceous. The oils of the Williston Basin exhibit a wide range of geochemical characteristics defined as "oil families", although the geochemical signature of the Cambrian Deadwood Formation and Lower Ordovician Winnipeg reservoired oils does not match any "oil family". Despite their close stratigraphic proximity, it is evident that the oils of the Lower Palaeozoic within the Williston Basin are distinct. This suggests the presence of a new "oil family" within the Williston Basin. Diagnostic geochemical signatures occur in the gasoline range chromatograms, within saturate fraction gas chromatograms and biomarker fingerprints. However, some of the established criteria and cross-plots that are currently used to segregate oils into distinct genetic families within the basin do not always meet with success, particularly when applied to the Lower Palaeozoic oils of the Deadwood and Winnipeg Formation.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

Design and analysis of generic LBS application developing platform

Location based services is promising due to its novel working style and contents.A software platform is proposed to provide application programs of typical location based services and support new applications developing efficiently. The analysis shows that this scheme is easy implemented, low cost and adapt to all kinds of mobile nework system.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that