脂蛋白(α)单克隆抗体对抗原决定簇的测定及应用研究

应田淋巴细胞杂交瘤技术,建立了4株抗人血清脂蛋白(α)(LP(a))杂交瘤细胞株(BH6、BH7、BH11和BH21),并以制备的单克隆抗体(McAb)对其抗原决定簇及免疫学特性进行了分析。Ig亚类测定:BH6为IgG1,BH7、BH11和BH21均为IgG2a,腹水效价为1×10~(-7)。特异性测定:LP(α)McAb与载脂蛋白(apo)AⅠ、AⅡ、B、CⅠ、CⅡ、CⅢ、E,人血清白蛋白,纤维蛋白溶酶原(pg)没有交叉反应。单抗相加试验和双抗体竞争试验结果证实,BH6和BH21为识别LP(α)上同一抗原决定簇的McAb;BH7和BH11则为针对LP(α)上另一抗原决定簇的McAb。分别利用单株和混合株McAb标记酶建立了可应用于人血清LP(α)含量测定的ELISA双抗体夹心法。     

抗人Apo B100单克隆抗体的制备及夹心ELISA检测方法的建立

目的:制备抗人载脂蛋白B100(Apo B100)单克隆抗体(mAb),建立人Apo B100双抗体夹心ELISA检测方法。方法:将人Apo B100抗原免疫小鼠,通过细胞融合、筛选后得到杂交瘤细胞株。将细胞株用无血清培养基扩大培养并纯化上清获得抗体,测定抗体亲和力、亚型、特异性及表位,最后建立双抗体夹心ELISA方法。结果:获得4株抗人Apo B100的杂交瘤细胞株(4-1-2、4-2-2、4-3-2、4-6-3),其分泌的抗体不与其他相关蛋白交叉反应,亲和力达到1×109L/mol。用4-3-2和4-6-3建立的双抗体夹心法的检测范围为(1.3～80)ng/mL,灵敏度1.24 ng/mL,批内变异系数均小于10%,批间变异系数均小于15%,回收率在90%以上。结论:成功制备了抗人Apo B100mAb,建立了定量检测人Apo B100的双抗体夹心ELISA方法,为Apo B100检测及疾病的诊断奠定基础。     

分泌抗HBeAg单克隆抗体杂交瘤细胞株的建立和初步应用

应用血清纯化的HBeAg免疫BALB/c小鼠,其脾细胞与Sp 2/0骨髓瘤细胞融合,获得两株分泌抗HBe的杂交瘤细胞株(HE_(20)IB_8、HE_(21)IIG_(11))。接种同系小鼠腹腔诱生的腹水中用ELISA测定抗HBe效价达3.2～6.4×10~6。因相捕捉HBeAg、HBsAg和HBcAg特异性试验以及中和试验证明是针对HBeAg的特异性抗体。临床应用作为ELISA检测HBeAg和抗HBe试剂中的包被抗体配合人抗HBe-HRP与Abbott试剂结果非常一致。     

抗重组人白细胞介素18单克隆抗体的制备与特性研究

目的 :获得有生物活性的小鼠抗重组人白细胞介素 18(rhIL 18)单克隆抗体。方法 :采用重组hIL 18免疫BALB C小鼠 ,应用杂交瘤技术 ,ELISA法筛选阳性杂交瘤细胞株并经多次克隆化。结果 :建立了 2株小鼠抗hIL 18单克隆抗体杂交瘤细胞 1C7和 1F5 ,染色体数目分别为 92条和 90条 ,所分泌的抗体分别为IgG2a和IgG1,轻链均为κ型 ,腹水IgG抗体经亲和层析法纯化后纯度达 95 %以上 ,效价为 1× 10 - 4和 1× 10 - 5,Westernblot显示 2株单抗均能特异性识别 18 3kD处的rhIL 18蛋白。 1C7单抗的亲和常数Ka=1 7× 10 5,1F5的亲和常数Ka=1 3× 10 5。 2株单抗识别不同抗原表位。结论 :制备的 2株单抗为进一步研究IL 18的分子结构、生物学功能及其与免疫相关性疾病的关系提供了实验材料。     

抗尿激酶单克隆抗体的研究——Ⅰ.分泌抗尿激酶单克隆抗体杂交瘤细胞系的建立

用市售的尿激酶(UK),免癌BALB/c小鼠。取脾细胞,与小鼠骨髓瘤细胞FO融合,获得六株分泌抗尿激酶单克隆抗体的杂交瘤细胞,分别命名为1UB_5、1UD_2、3UA_2、3UD_(12)、3UG_2、3UH_7。其中,1UB_5及1UD_2连续传代培养4个月以上,其培养液抗体的ELISA效价仍为10~(-2)～10~(-3)。两株细胞经扩大培养后,注入C_(57)×BALB/c杂交后的F_1代小鼠,诱生的腹水抗体效价为5×10~(-6)～10~(-7)1UB_5、1UD_2所分泌的抗体经ELISA鉴定,分属IgG_1和IgG2a。用高纯度的低分子量UK包被,ELISA检测后表明,两株细胞所分泌的为抗高分子量UK的McAb。 抗UK的McAb用于粗制品UK的纯化及监察某些肿瘤发生与发展的进一步工作正在进行之中。     

小鼠抗人C5aR短肽（9—30）单克隆抗体制备及鉴定

目的获得有生物功能的小鼠抗人 C5 a R短肽单克隆抗体。方法对 C5 a受体 (CD88)二级结构和 B细胞表位进行分析研究 ,采用 Fmoc方案固相合成 C5 a R N-端第 9～ 30位氨基酸残基的 2 2 -肽 ,以此为抗原免疫 Balb/ c小鼠。结果建立了 1株小鼠抗人 C5 a R短肽杂交瘤细胞系 E3,其平均染色体数目为 10 2条 ,所分泌抗体为 Ig G1,κ型 ,腹水抗体效价为 1×10 - 4 ～ 1× 10 - 6 ,可识别 U 937、人脐静脉内皮细胞 (VEC)和 PMN等表达 C5 a R细胞。 E3单抗的亲和常数 Ka=2 .5× 10 5 ,其结合表位为 C5 a R第 15～ 2 1位氨基酸基序 D1 5 DKDTL2 0 D。结论 B细胞表位多肽具有免疫原性 ,可制作单克隆抗体 ,为 C5 a-C5 a R相互作用的研究以及 AL I、ARDS等 C5 a相关疾病的研究提供实验材料。     

抗人CA15-3单克隆抗体的制备、鉴定及化学发光检测体系的建立

目的获得抗人CA15-3单克隆抗体(mAb),建立CA15-3双抗体夹心化学发光检测体系。方法人CA15-3糖抗原免疫小鼠后,通过细胞融合,筛选到阳性杂交瘤细胞。杂交瘤细胞扩大培养后,纯化获得抗CA15-3的mAb。对抗体进行纯度、效价、表位和亚型的鉴定并建立双抗体夹心检测CA15-3化学发光体系,对该体系进行准确度、最低检测限、线性、重复性与特异性评估。结果获得5株阳性信号较强的杂交瘤细胞(分别为#3-1-3﹑#5-2-2、#11-2-2、#12-1-3和#16-1-3)。其分泌的抗体效价均大于10-8g/mL,轻链均为κ链,重链#3-1-3为IgG2a、#5-2-2与#12-1-3为IgG2b、#11-2-2与#16-1-3为IgG3。双抗体夹心体系在(0~300)U/mL范围内线性关系良好,其准确度的回收率为97.45%;最低检测限为0.59 U/mL;线性相关系数为0.9978;低高值的变异系数(CV)均小于10%;与肿瘤标志物AFP、CEA、CA50、CA19-9和CA72-4均不交叉。结论成功制备了抗人CA15-3 mAb,建立了双抗体夹心检测CA15-3化学发光体系。     

抗人补体膜攻击复合物新抗原单克隆抗体的制备及鉴定

目的　研制针对人补体膜攻击复合物 (MAC)新抗原的特异性单克隆抗体。方法　在兔红细胞表面组装人MAC ,分离纯化RBC膜蛋白。以初步纯化的MAC免疫Balb c小鼠 ,取免疫小鼠脾细胞与SP2 0骨髓瘤细胞融合 ,采用免疫斑点杂交的方法分别以纯化的单一补体成分C5、C6、C7、C8、C9和MAC同时进行阴性和阳性筛选。结果　获得了 2株仅与MAC反应而不与各单体成分反应的单克隆抗体。进一步以体外组装的补体终末复合物进行鉴定 ,证实所获单克隆抗体可特异性识别MAC复合物中C9分子暴露出的新抗原。经ELISA法测定 ,2株杂交瘤细胞的培养上清液和腹水效价分别为 1× 1 0 -3 ,1× 1 0 -3和 1× 1 0 -6,1× 1 0 -7;单克隆抗体的重链均属小鼠IgG1亚类 ,轻链均为κ型。结论　获得了 2株特异性识别MAC新抗原的单克隆抗体。     

抗HBsAg表位单克隆抗体的制备及HBV血清型夹心ELISA的建立

目的制备抗乙型肝炎病毒(HBV)表面抗原(HBsAg)表位单克隆抗体(mAb),建立HBV血清型夹心ELISA检测方法。方法将HBsAg亚型蛋白分别免疫小鼠,通过细胞融合、高通量筛选ELISA(HTS-ELISA)筛选后得到杂交瘤细胞株并在无血清培养基中扩大培养。采用蛋白A对抗体进行纯化并测定抗体亲和力、亚型、特异性,最后建立双抗体夹心ELISA。结果HBsAg的亚型蛋白ad、adr及ayw分别免疫小鼠后,其血清效价达到1∶105。细胞融合及HTS-ELISA筛选后获得4株杂交瘤细胞株。经过纯化后得到的mAb分别命名为#2-4,#18-3,#7-7,#56-5,亲和力都在1×109L/mol以上。间接ELISA结果显示,#2-4,#18-3,#7-7,#56-5分别特异性地与HBsAg的"d,y,r,w"表位结合。用抗HBsAg"a"表位的mAb#3-11分别与这4株抗体组合,进行夹心ELISA检测。结果显示,不同的ELISA组合能特异性地检测HBsAg的"d,y,r,w"表位。结论成功制备了亲和力高、特异性好的抗HBsAg 4个表位的mAb,建立了检测HBV血清型的双抗体夹心ELISA。     

抗人CA19-9单克隆抗体的制备和鉴定及化学发光检测体系的建立

目的制备抗人糖抗原19-9(CA19-9)单克隆抗体(mAb),建立双抗体夹心检测CA19-9化学发光体系。方法 CA19-9免疫小鼠后,通过细胞融合技术,获得抗CA19-9的mAb。鉴定其抗体的纯度、效价、特异性和配对,建立双抗体夹心化学发光体系。在包被缓冲液、包被浓度及加样模式上优化化学发光体系,对该体系进行准确度、最低检测限、线性、重复性评估以及血清样本的检测。结果获得4株名称为#1-1、#2-1、#3-1和#4-1的mAb。抗体效价均大于10-8,与CA125、CA15-3、CA72-4均无交叉反应。其中#3-1 mAb与#2-1-HRP建立的双抗体夹心体系经优化后,其检测范围为0~1 000 U/mL;最低检测限为2.0 U/mL;线性相关系数为0.9999。与罗氏试剂检测结果对比,Kappa系数大于0.75;相关系数大于0.9。结论成功制备了抗人CA19-9 mAb,建立了双抗体夹心检测CA19-9化学发光体系。     

Level of functioning of siblings and parents of probands of varying degrees of retardation

A further analysis of already published data supports the position that retardates of low ability level less frequently have retarded siblings, retarded parents, and parents low in occupational level than do retardates higher in ability level. The analysis supports the position that there are two types of retarded individuals, persons retarded as a result of gene or chromosomal anomalies, brain injury, etc., who more frequently occur in the lower-level retardate group, and persons whose retardation represents polygenic segregation, who more frequently occur in the higher-level group.     

Assessment of the efficacy of treatment of hemangiomas

Assessment of the aftereffects of cryoexposure and ultrahigh-frequency cryoexposure on hemangioma tissue of various types, cavernous and squamous, showed a higher cryogenic effect in hemangiomatous tissue preexposed to ultrahigh-frequency waves. A quantitative criterion is proposed for assessing the efficacy of the studied methods of exposure. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 6, pp. 669–672, June, 1995     

侧脑室注射链脲佐菌素对大鼠海马神经元突触的影响

为观察链脲佐菌素(streptozotocin,STZ)双侧侧脑室注射对大鼠海马神经元突触的影响,本研究将Wistar大鼠随机分为对照组和模型组,模型组大鼠分别于第1、3d双侧侧脑室重复注射STZ3mg/kg,对照组以人工脑脊液代替STZ。21d后,取大鼠海马,免疫组织化学染色及Western blotting方法观察突触素、活性调节细胞骨架相关蛋白(activity-regulated cytoskeletal-associatedprotein,Arc)的表达;电镜观察海马CA1区神经元突触超微结构的改变。结果显示:与对照组相比,模型组大鼠海马内突触素蛋白表达显著减少,而Arc蛋白表达显著增多;模型组海马CA1区神经毡内突触结构异常,突触小泡聚集增多。以上结果提示:脑室注射STZ可影响大鼠海马突触相关蛋白的表达,引起突触超微结构异常,干扰了神经元突触信号的传导。     

Effect of conditions of culture on quantitative assessment of viability of living tissues

The viability of tissue explants of mouse kidneys, estimated quantitatively by growth in plasma-free culture, was shown to depend on several factors connected with the conditions of culture. The effect of methods of treatment of the serum, pH of the medium, and the character of distribution of the tissue explant during culture, the size of the explants, and the firmness of attachment of the tissue fragments to the substrate on this index was demonstrated.Problem Laboratory, Arkhangel'sk Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 11, pp. 629–631, November, 1978.     

社区精神分裂症病人照料者的生活质量研究

通过对９３名精神分裂症患者的家庭照料者与１００名对照者比较研究表明，照料者的躯体功能、心理功能、社会功能与经济状况都显著差于对照者。照料者的生活质量与被照料者的病期、病情严重度、整体功能、药物副作用有关；同时，照料者身份为患者母亲者其生活质量受影响最大。     

膝关节骨性关节炎的关节镜治疗体会

目的我科自2001～2005年4月对59例67膝的骨性关节炎（0A），进行关节镜检查及镜下清理术。方法镜检：膝关节骨性关节炎伴不同程度滑膜炎，滑膜皱壁粘连纤维片，关节软骨不同程度破坏。镜下清除增生滑膜，松解粘连，去除剥脱软骨、修复关节面。结果随访2月～4年，平均1.5年，术后综合评估2月～1年，优良率94％；1～2年，优良率79.2％；2年以上优良率54.8％。结论关节镜对膝关节骨性关节炎诊断能提供了比较全面的资料，并对骨性关节炎早期有良好疗效，具有创伤小、恢复快、并发症少和重复治疗等优点。     

Evaluation of fungicidal activity of extracellular filtrates of cyanobacteria--possible role of hydrolytic enzymes

A set of seventy axenised and unicyanobacterial isolates belonging to the genus Anabaena were evaluated for biocidal activity against a set of phytopathogenic fungi. Among them, 35 Anabaena strains showed zone of inhibition against one or more fungi. The extracellular filtrates from 4 and 8 weeks old cultures of these Anabaena strains were further evaluated in terms of hydrolytic enzymes, proteins and IAA employing standard methods. Significant differences were also observed among the strains in terms of their FPase, chitosanase and xylanase activity, while low and relatively similar values of CMCase, cellobiase and protease activity were recorded in the strains analyzed. IAA production was also observed in all the strains. Comparative evaluation of activity of hydrolytic enzymes and antifungal activity revealed that such enzymes may contribute to the fungicidal activity of the cyanobacterial strains, besides other bioactive compounds, including IAA, which are established promising traits for biocontrol agents. This study is a first time report on the production of hydrolytic enzymes by these oxygenic photosynthetic prokaryotes, which can be potential candidates for the development of biocontrol agent(s) against selected phytopathogenic fungi.     

Pathways of the realization of immunomodulating effects of neurotransmitters

Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 115, N ^o 4, pp. 401–404, April, 1993     

Characterization of sterols of three digenetic trematodes of buffalo

Sterols of three digenetic trematodes were isolated and characterized by infrared and 1H nuclear magnetic resonance spectroscopies, and gas-liquid chromatography and gas chromatography-mass spectrometry. Sterols identified were cholesterol, cholestanol, 24-methylcholesterol, 24-methylcholestanol, 24-ethyl-22-dehydrocholesterol, 24-ethyl-22-dehydrocholestanol, 24-ethylcholesterol and 24-ethylcholestanol.     

Mechanisms of generation of slow components of bioelectric activity in diagnostics of functional state of the gastrointestinal tract

New electrophysiologic devices were used for the diagnostics of the state of excitable structures in the gastrointestinal tract and correction of their motor functions. Bioelectrical and biomechanical activities form the basis of functioning of internal organs. The mechanisms of generation of slow bioelectrical activity that are important for clinical and physiological studies are described. One of these mechanisms is a capacitance parametric transducer converting the energy of contractions into specific electric signals reflecting muscle functions. Another mechanism results from slow oscillations of resting potentials of interrelated excitable cells in large neuromuscular structures of internal organs. The elaborated procedure is efficient for preventing early postoperative paresis of the gastrointestinal tract. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 10, pp. 448–452, October, 1999