~(125)碘-绒电膜促性腺激素的制备与鉴定

作者制备了~(125)碘-绒毛膜促性腺激素,作为该激素受体放射分析的示踪物。结果显示,~(125)碘际记激素具有免疫活性及生物活性,可用于受体放射分析。对~(125)碘标记条件作了讨论。     

胰岛素的碘标记对抗胰岛素抗体免疫活性的影响

本文用原型胰岛素和A14“冷”单碘原子碘化胰岛素作标准品,同时用两株不同的豚鼠抗胰岛素抗血清作放射免疫分析。其结果经Scatchard分析和比较标准曲线斜率的方法证明,胰岛素分子A链第14位酪氨酸的碘标记,干扰其中一株抗血清(D7)固有的抗原抗体结合,降低其免疫活性,而对另一株抗血清(GP12)没有影响。此现象的意义在于进行自身置换分析等特殊的放射免疫分析时,必须考虑到~(125)碘标记对免疫活性或生物活性的干扰问题。     

[~(125)碘]单碘胰岛素的标记

激素受体的研究需要既有免疫活性,又有完好的生物活性的标记激素,70年代中期,国外开始研究[~125碘]单碘胰岛素的标记。1979年,Linde等用乳过氧化酶法标记,应用长柱聚丙烯酰胺凝胺圆盘电     

A14-或A19-[~(125)碘]单碘原子碘化胰岛素的制备及其意义

本文介绍了用聚丙烯酰胺长柱凝胶园盘电泳,制备A14-及A19-[~(125)碘]单碘原子碘化胰岛素的方法,并用放射免疫法对标记物的质量作了鉴定。结果显示,这两种胰岛素单碘标记物的免疫活性和贮存有效期均高于国产的~(125)碘标记胰岛素。文中对胰岛素单碘标记物在放免分析中的意义进行了讨论,并着重阐述了标记物与抗体之间的关系。     

肝细胞生长因子生物活性的体外检测的研究

测定了狗的再生肝中肝细胞生长因子（ＣＨＧＦ）不同剂量对肝细胞增殖的刺激作用，发现其与肝细胞增殖的增强有剂量依赖关系；同时，比较了ＣＨＧＦ在－２０℃贮存六个月前后的生物活性，发现－２０℃贮存六个月后，仍保持生物活性；并对ＣＨＧＦ与乳猪肝的肝细胞生长素（ＰＨＧＦ）的生物活性进行了比较，发现二者生物活性基本一致，ＣＨＧＦ的生物活性略高于ＰＨＧＦ的生物活性。     

高比活度单克隆抗体放射性碘标记方法的探讨

本文采用N-溴代琥珀酰亚胺(N-bromosuccinimide,BSI)作为碘化试剂,对单克隆抗体WuT9进行高比活度的碘标记,碘利用率介于84％～95％,平均90％;免疫活性分数介于57％～70％,平均64％,48h培养未见微生物生长;于标记后3d内,标记单克隆抗体结构稳定,在荷瘤小鼠及部分临床病例中试用获得满意结果。     

麝香草叶中酸性多糖的精制和抗补体活性

麝香草Thymus vulgaris L.在南欧、西亚和北非均有栽培,其甲醇提取物有多种生物活性,但主要作为食用的热水提取物的生物活性未见报道。作者发现该植物叶热水提取物具抗补体活性,并对其进行了精制和鉴定。该植物用甲醇于室温提取5次,过滤后残余物再用热水提取。热水提取物(TV-O)分别用30％、75％和95％乙醇沉淀。75％乙醇     

库存全血抗凝血酶Ⅲ及蛋白C变化的研究

对15名健康献血员4℃库存血进行了抗凝血酶Ⅲ活性及蛋白C抗原含量的动态观测。结果发现，抗凝血酶Ⅲ活性在采血后12h内活性即明显下降，在贮存3d时活性下降超过50％，在贮存7d时失去70％左右的抗凝活性，蛋白C抗原含量较稳定，在贮存第4d开始下降，贮存第30d下降才达50％以上。因此认为，在临床补充抗凝成份替代疗法中，输注贮存3d以内血液有可能补充一定量的抗凝血酶Ⅲ，而1月以内的贮存血可补充蛋白C。     

库存全血抗凝血酶Ⅲ及蛋白C变化的研究

对１５名健康献血员４Ｃ库存血进行了抗凝血酶Ⅲ活性及蛋白Ｃ抗原含量的动态观测。结果发现，抗凝血酶Ⅲ活性在采血后１２小时内活性即明显下降，在贮存３天时活性下降超过５０％，贮存７天时失去７０％左右的抗凝活性，蛋白Ｃ抗原含量较稳定，在贮存第４天开始下降，贮存第３０天下降才达５０％以上，因此认为，在临床补充抗凝成份替代疗法中，输注贮存３天以内血液有可能补充一定量的抗凝血酶Ⅲ，而１月以内的贮存血可补充蛋白Ｃ。     

抗TSH独特型抗体与TSH及其受体的关系

以TSH单克隆抗体免疫7只新西兰家兔，对免疫后家兔中抗TSH独特型抗体及其对家兔甲状腺功能的影响进行了观察。免疫25周后，家兔的T4水平和甲状腺摄碘率（TRUR）较免疫前显著降低（T416．17±5．69nmol／L比30．11±15．24nmol／L，P＜0．05），2h和4hTRUR免疫前后的差值分别为11．44％±10．38％和14．21％±12．50％（P均＜0.05）。以抗TSH单克隆抗体亲和层析柱分离免疫后兔血清，发现免疫后家兔产生TSH的抗独特型抗体和抗抗独持型抗体，以TRAb放射受体分析系统和TSH结合物的固相竞争，确认上述二种抗体分别模拟TSH和抗TSH抗体，它们对家兔的甲状腺功能可产生不同的影响。     

~(125)I-绒毛腺促性腺激素的生物活性研究

作者对自制的~(125)I-HCG的生物活性作了受体亲和力、靶组织专一性及受体对标记物的有限结合能力等试验,结果表明:~(125)I-HCG具有良好的生物活性。根据受体-配体结合理论及本研究结果,提出了测试~(125)I-HCG生物活性的4个可行指标。     

绒毛膜促性腺激素受体单克隆抗体的制备及其特性的初步研究

以纯化嗜麦芽假单胞菌的绒毛膜促性腺激素(CG)受体为抗原,融合得到4株(ED490,DG390,AB890,GE590)对细菌CG受体特异的单克隆抗体杂交瘤细胞,其腹水和培养上清中的抗体滴度分别为10~(-2)～10~(-6)和1～10~(-2);亚类鉴定显示:单抗GE590为IgGl,单抗ED490、DG390和AB890为IgG2b。研究发现,ED490、AB890和DG390与细菌CG受体作用后,~(125)I-HCG仍能结合该受体,揭示其作用于受体不同位点。GE590与CG受体结合实验发现,随着单抗浓度不断增加,~(125)I-HCG与受体结合逐渐减少,推测该单抗与~(125)I-HCG结合受体同一位点。     

绒毛膜促性腺激素受体单克隆抗体的制备及其特性的初步研究

We reported the production of monoclonal antibodies (McAbs) against chorionic gonadotropin hormone (CG) receptor by fusing spleen cells of BALB/c mice which had been immunized by purified bacteria (Pseudomonas maltophilia) CG receptor with mouse myeloma line SP2/0. Four hybridoma cell lines secreting CG receptor McAbs were obtained (ED490 DG390, AB890 and GE590). The titers of specific antibodies of both mice ascites and culture supernatant were 10(-2)-10(-6) and 1-10(-2) respectively, by solid phase ELISA. Double-Immunodiffusion test showed that the McAb GE590 was IgG1, and the McAbs ED490, DG390 and AB890 were IgG2b Immunoprecipitation indicated that 125I-HCG could bind the HCG receptor which had reacted with McAbs ED490, AB890 and DG390, suggesting that they may recognize the receptor with different antigenic determinant. Interaction of McAb GE590 with the receptor showed that the increased concentration of GE590 was in inverse proportion to the amount of 125I-HCG binding receptor, indicating that both the McAb and 125I-HCG could recognize a common site of receptor and that increased concentration of McAb GE590 may induce some change in conformation and structure of the receptor. Our study suggested that these McAbs may be used for studying structure of CG receptor.     

人的正常卵巢及巢癌绒毛膜促性腺激素受体的研究

Human Chorionic Gonadotropin (HCG) is major physiological luteotropic factors for the human corpus luteum. The observations strongly suggest that the human ovary possesses a gonadotropin receptor in the cell membrane. We studied the HCG receptor in normal human ovary and ovarian tumors. Twenty-three human ovarian specimens and 16 ovarian tumor specimens were obtained from women patients having gynecological surgery. Ovaries were homogenized and sonicated. The homogenates were centrifuged at 2000 g for 15 min. After sucrose density gradient ultracentrifugation (78,000 g, 4 h), two fractions were collected from layer of 33% and interface between 33% and 37%. Thirty micrograms of ovarian protein, 8 ng 125I-HCG and unlabeled HCG in a final volume of 0.5 ml of 0.05 mol/L Tris buffer were incubated at 30 degrees C for 2 h. The results were shown in the table.     

咪唑并吡啶类外周苯二氮卓受体配体的合成新方法

目的咪唑并[1，2-a]吡啶类PBR配体新的合成方法及其同位素^125I-标记。方法非放射活性的咪唑并[1，2-a]吡啶通过5-氯-2-氨基吡啶与溴代酮酯缩合后再酰胺化而得。起始溴代酮酯经付-克酰化、酯化和溴代而被制备。放射标记的^125I-咪唑并[1，2-a]吡啶通过Na^125I在氯胺T反应体系中经碘脱锡交换反应被制备。由反向HPLC纯化得^125I-咪唑并[1，2-a]吡啶。结果非放射活性的咪唑并[1，2-a]吡啶6步合成总收率25．2％，放化得率68％，放化纯度大于98％。结论咪唑并[1，2-a]吡啶类PBR配体的合成和标记方法简便，且回收率高。     

A prospective study: intraoperative 125I radioactive seed implant therapy in advanced esophageal squamous cell carcinoma

Objective: To investigated the role of intraoperative iodine-125 (125I) brachytherapy as a treatment option for advanced thoracic esophageal squamous cell carcinoma (ESCC). Methods: Using preoperative computed tomography (CT)-based staging criteria, between 2000 and 2008, 298 patients with ESCC (stage II-III) were enrolled in this prospective study. With informed consent, patients were randomized into two groups: intraoperative 125I seed implantation and surgery alone (control group). Twenty to forty 125I seeds (0.5 mCi per seed), with a total activity in 10~30 mCi, and a matched peripheral dose (MPD) of 60~70 Gy, were implanted under direct visualization. The surgical procedure used in this study was either a radical resection, which involved an esophagectomy through a left thoracotomy with two-field lymphadenectomy, or palliative resection. The postoperative complications were observed and recorded. The location and quality assessment of 125I seeds were assessed using CT scans or X-ray imaging. The short-term efficacy was evaluated according to WHO criteria. The 1, 3, 5 and 7-year survival rates were determined on follow-up. Results: There was no displacement or loss of 125I seeds. The local recurrence rates in the intraoperative 125I seed implantation group and control group were 14.9% and 38.7%, respectively (P < 0.05). An objective response rate of 92% was observed in the seed implant group, which was significantly higher than 0% in the control group (P < 0.05). There was no significant difference between the two groups when comparing of complications (P > 0.05). The 1-year survival rate of the two groups were not significantly different (P > 0.05). However, the 3, 5 and 7-year survival rates in the united 125I group (64%, 55.3% and 8%, respectively) were statistically different from those in the control group (52%, 29.1% and 1.4%,respectively)(P < 0.05). Conclusion: Intraoperative 125I seed implantation is safe and effective for advanced ESCC. Seed implantation may reduce the local recurrence rate and improve survival in patients with ESCC. The MPD of 60~70 Gy, with single 125I seed activity of 0.5 mCi, is reasonable.     

血清BDNF和CA125水平对急性心肌梗死患者病情及预后的评估价值

目的分析血清脑源性神经营养因子(BDNF)和糖类蛋白125(CA125)水平对急性心肌梗死(AMI)患者病情及预后的评估价值。方法选取2016年12月-2018年6月北京中医药大学孙思邈医院心血管内科收治的AMI患者136例作为研究组,选取同期门诊健康体检正常者50例作为健康对照组。采用酶联免疫吸附法检测血清BDNF水平,电化学发光免疫分析法检测CA125水平。随访1年记录不良心血管事件发生情况,比较不良心脏事件亚组和无不良心脏事件亚组患者血清BDNF和CA125水平。采用Logistic回归分析急性心肌梗死患者预后影响因素。结果研究组心率及血清BDNF、CA125、c-TnI、BNP水平均高于健康对照组(t=10.196、22.142、12.302、7.301、49.705,P均<0.001)。随访1年发生心脏不良事件38例(27.94%),不良心脏事件亚组患者血清BDNF、CA125水平均高于无不良心脏事件亚组患者(t=3.258、4.496,P均<0.001)。BDNF预测AMI患者1年不良心血管事件发生率的ROC曲线下面积(AUC)为0.871,当BNDF截点值为3.87 ng/ml时预测AMI患者1年不良心血管事件敏感度为84.1%,特异度为82.9%,约登指数为0.670。CA125的AUC为0.746,当CA125截点值为42.54 U/ml时预测AMI患者1年不良心血管事件的敏感度为83.2%,特异度为81.5%,约登指数为0.647。Logistic回归分析结果显示,高龄(OR=1.89,95%CI 1.02~3.51)、心率快(OR=1.77,95%CI 1.06~2.95)、高水平BDNF(OR=4.76,95%CI 2.38~9.56)、高水平CA125(OR=3.49,95%CI 1.71~7.12)、高水平cTnI(OR=3.80,95%CI 1.654~8.750)、高水平BNP(OR=4.90,95%CI 2.23~10.77)为AMI后不良心血管事件的危险因素。结论 AMI患者血清BDNF、CA125水平明显增高,且高水平BDNF、CA125是AMI患者发生不良预后的风险因素,提示二者可作为评估AMI患者病情及预后的预测因子。     

Characterization of the human DNA polymerase delta catalytic subunit expressed by a recombinant baculovirus

The catalytic subunit of human DNA polymerase (pol) delta, p125, was expressed in recombinant baculovirus-infected insect cells, separated from a baculovirus-encoded DNA polymerase, and was purified to homogeneity by affinity trapping with a histidine-octapeptide at the C-terminus of p125 as the ligand. Purified p125 showed DNA polymerase activity resembling conventionally purified calf thymus pol delta. However, the two differed in four ways: 1) the specific activity of recombinant p125 was one quarter of the calf thymus pol delta; 2) the recombinant p125 was relatively resistant to aphidicolin; 3) the apparent Km for dTTP of the recombinant p125 was estimated at 33 microM, 15-fold the value for calf thymus pol delta; and 4) the recombinant p125 was not stimulated by recombinant PCNA, while activity of calf thymus pol delta increased 150-fold in response. Furthermore, PCNA did not stimulate either the p125 incubated with p50, a small subunit of pol delta, or co-expressed with p50 in insect cells. The full length recombinant p125 migrated slightly faster than pol delta from human cell lines, Jurkat or HeLa, upon SDS-polyacrylamide gel electrophoresis, suggesting a post-translational modification. The results indicate that in vivo assembly of the fully active complex of pol delta requires factors in addition to p125 and p50 subunits, and/or a post-translational modification of p125.     

重组人前列腺特异性抗原纯化与活性鉴定

目的：纯化和鉴定重组人前列腺特异性抗原（PSA）,为PSA 的生物学特征研究、临床检测和治疗应用提供关键材料。方法： 采用阳离子交换层析技术纯化重组PSA（rPSA）,利用Western blotting 鉴定其特异性,利用PSA底物S-2586检测rPSA活性。结果：最适甲醇诱导浓度为1.5%,经鉴定纯化的rPSA为人PSA。在不同反应条件下,在3.5 h内酶活性与时间成正比,rPSA 活性显著高于正常精浆PSA酶活性（P<0.05）,酶活性的最适温度为30℃。结论：成功纯化出具有酶活性的rPSA。     

优立新与西力欣序贯疗法治疗下呼吸道感染的对照研究

本试验旨在评价优立新序贯疗法治疗下呼吸道感染的临床疗效和安全性。西力欣作为对照。入选病例６１例,其中试验组（优立新）３１例,对照组（西力欣）３０例。给药方法采用静脉注射随后口服给药的序贯疗法：静脉注射优立新３～１２ｇ／天,连续 ３～７天,继之口服舒他西林３７５ｍｇ～７５０ｍｇ,每天２次,连续５～７天;静脉注射西力欣３～６ｇ／天,继之口服新菌灵２５０ｇ～５００ｇ,每天 ２次,疗程相同。结果试验组和对照组的临床有效率分别为８０．６％（２５／３１）和９０％（２７／３０）。６１例患者共分离出致病菌４１株。治疗结束后两组的细菌清除率分别为 ８２．１４％（２３／２８）和 ６９．２３％（９／１３）,毒副作用反应发生率分别为 ６．４５％和３． ３３％。上述结果经统计学处理无显著性差异。无病者因毒副作用而退出试验。