人血清载脂蛋白A-I的分离、纯化及其测定

用超离心法制备人血清高密度脂蛋白(HDL);甲醇/氯仿脱脂;经Sephadex G-150层析及DEAE-纤维素(DE-52)离子交换得纯化载脂蛋白A-I(Apo A-I),免疫新西兰兔,获得效价为1:32的特异性抗人Apo A-I抗血清。用本室建立的单向火箭免疫电泳法测定了158名健康中年人与9名老人的血清Apo A—I含量,结果发现:β-脂蛋白<5g/L,总胆固醇<2mmol/L的人血清Apo A-I含量男女分别为2.06±0.16g/L和2.07±0.15g/L;而β脂蛋白≥5g/L,总胆固醇≥2mmol/L的人血清Apo A-I含量男女分别为1.75±0.15g/L和1.82±0.1g/L,由此可见,血脂正常的男性和女性的Apo A—I含量均显著高于血脂高的男性和女性。80岁以上老年人的血清Apo A-I含量为2.1±0.1g/L,也在正常范围。与文献报道比较,我国人的血清Apo A-I含量高于外国人,这与我国人HDL水平较外国人高一致。     

沙培林增强人腹腔巨噬细胞抗癌免疫功能的研究

目的:探讨腹腔内注射沙培林增强人腹腔抗癌免疫功能的机制。方法:72例早中期胃肠道肿瘤患者术前48h和24h腹腔内分别注射生理盐水和5KE的沙培林,术中采集腹腔内巨噬细胞,计数并测定乳酸脱氢酶(LDH)和酸性磷酸酶(ACP)的活性,巨噬细胞吞噬活力,一氧化氮(NO)的分泌以及对人胃癌MKN1细胞的细胞毒性进行分析。同时采集大网膜,对大网膜乳斑的数量和面积进行观察。结果:沙培林显著增加腹腔巨噬细胞(PMΦ)的数量和NO的分泌,增强LDH和ACP的活性,吞噬活力,以及抗癌细胞毒性,也显著增加了大网膜乳斑的数量和面积。结论:腹腔内注射沙培林可显著增加人大网膜乳斑的数量和面积,并因此增加PMΦ的数量,增强PMΦ的活性。因而增强了腹腔巨噬细胞的免疫功能。     

Safety, tolerability and pharmacokinetic study of recombinant human parathyroid hormone in healthy male Chinese subjects

AIM： To determine the safety, tolerability and pharmacokinetic parameters of a new drug recombinant human parathyroid hormone [ rhPTH （1-84）] in healthy male Chinese subjects. METHODS： domly divided Thirty-six healthy male volunteers were rangroups received into 3 groups. The volunteers in these single subcutaneous injection of rhPTH （ 1-84） in a dosage of 1, 2 and 4 μg/kg respectively. Blood samples were obtained before and after administration within 24 hours. The rhPTH concentrations in sennn were determined by enzyme linked immunosorbent assay （ELISA）. The pharmacokinetic parameters determined with use of standard noncompartmental analysis were the maximum serum concentration （ Cmax ）, the time to attain that concentration （ tmax ）, and the area under the serum concentration-time curve up to 24 hours（ AUC0-24 ） and up to infinity （AUC0-∞）. Dose proportionality of pharmacokinetic parameters （AUC, Cmax of every volunteer of each dosage and A UC was computed from log transformed data） and was examined by mean of analysis of variance （ANOVA） using SPSS software package. In the study, subjects＇ symptoms, objective signs, and vital signs, including blood pressure, heart rate, respiratory rate and body temperature, were checked and 12-lead electrocardiography was recorded before and after drug administration within 24 hours. Routine laboratory tests, including hematology, blood biochemistry, serum electrolyte, and urinalysis, were performed before and after drug administration within at 24 hours.[第一段]     

As_2O_3诱导人肝癌细胞凋亡及对Fas表达和钙含量的影响

化疗在原发性肝癌的综合治疗中占重要地位.为寻找肝癌治疗新的有效药物,我们观察了As2O3对人肝癌细胞株BEL7402的生物学效应,以期为As2O3临床治疗肝癌提供实验依据.     

Comparative studies of chemotherapy of human tumor cells in vitro by tritiated thymidine uptake inhibition and soft agar clonogenic assay

A rapid nonsterile short-term assay has been investigated in which percent inhibition of incorporation of the DNA precursor (3H)-thymidine is measured following exposure of tumor cells to the test drugs. To evaluate the usefulness of the short-term assay in providing a rapid reliable assessment of chemotherapeutic response, the short-term assay was compared with the soft agar clonogenic assay. Sensitivity to five anticancer drugs was compared using three human tumor cell lines (epidermoid carcinoma of the oral cavity, pancreatic carcinoma, and bladder carcinoma). The short-term assay produced results that were similar to results of the clonogenic assay in two of the three tumors tested, for three drugs (cis-platin, doxorubicin, and BCNU), but did not detect responses to two antimetabolites (5-FU and MTX) in any tumor. Further studies of this short-term assay should focus on alkylating agents and other nonantimetabolites.     

Reassessment of the accuracy of traditional sperm characteristics and adenosine triphosphate (ATP) in estimating the fertilizing potential of human semen in vivo

Receiver operating characteristic curves and accuracy parameters were computed for traditional sperm characteristics (concentration, motility, morphology) and the number of peroxidase negative cells, and the concentration of adenosine triphosphate (ATP) in semen from populations of fertile and infertile men, and men who achieved a pregnancy after varicocele treatment. The percentage and concentration per millilitre of spermatozoa with rapid linear progressive motility, and the ATP concentration, provided the best discrimination between fertile and treated fertile from infertile men. The misclassification rate was higher for sperm morphology, total progressive motility and viability, whereas sperm concentration and the total sperm count per ejaculate had the worst discriminating power. The number of peroxidase negative cells per 100 spermatozoa was highly specific in identifying men who achieved pregnancy after varicocele treatment. The lower limit of normality of sperm characteristics was remarkably different between fertile men and men achieving pregnancy after treatment or during infertility work-up.     

雷公藤内酯对人胃癌细胞株FGC_(85)杀伤作用的光镜和电镜观察

本文应用核苷掺入技术及电镜观察首次证实雷公藤内酯对人胃癌细胞株 FGC_(85)的杀伤作用。用药早期,细胞数,分裂指数及 DNA,RNA 合成无明显变化,但出现核仁脱粒及核仁破碎等变化;晚期,电镜观察发现细胞以凋落方式死亡,药物主要作用于间期细胞,其杀伤机构的始动环节可能与核酸代谢障碍无关。     

Quantitative observation by enzyme cytochemistry of human rectal adenocarcinoma cells killed by LAK cells

Activity of succinic dehydrogenase(SDH)and acid phosphatase(AcPase)of effector-target cells during the process of LAK cells killing HR8348 cells was estimated by enzyme cyto-chemistry technique.SHD positive granules and AePase gray level were assayed with MIAS-300image analyser.The results showed:(1)After cocultivation of effector and target cells for vari-ous times,the activity of AcPase of HR8348 cells was apparently higher than that of the controlgroup,and it increased following prolonged coincubation.SDH activity of target cells increasedmarkedly within 30 and 60 rain cocultivation,but became low after 90 min treatment.(2)Ac-Pase content within LAK cells at 60,90,120,180 and 240 rain cocultivation was significantlyhigher than that of control group(P<0.01).The phenomenon of high AcPase and SDH activitywithin effector-target cells indicates that the function of the two types of cells was in an activestate.At the early stage of effector-target combining,the increase of SDH with HR8348 cellsmay be related to defensive function of the target cells.Higher AcPase activity of target cells in-dicates the activation of lysosomal enzyme which serves as the material basis for autolysis of thecells.     

Matrix Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-1 in the Retinal Pigment Epithelium and Interphotoreceptor Matrix: Vectorial Secretion and Regulation

Matrix metalloproteinases (MMPs) and the tissue inhibitors of metalloproteinases (TIMPs) play an essential role in both normal and pathological extracellular matrix degradation, and a TIMP has been associated with at least one type of retinal degeneration. We have studied expression of MMP-2 and TIMP-1 by zymography, immunocytochemistry, and immunoblotting in the retinal pigment epithelium (RPE) from normal, aged and diseased retinas. MMPs and TIMPs were found in the rat RPE, interphotoreceptor matrix (IPM), and in media conditioned by human and rat RPE in culture. In other polarized cells, MMPs and TIMP-2 are secreted vectorially towards the basal lamina. In the RPE, however, MMP-2 and TIMP-1 were secreted preferentially from the apical surface, the surface bordering the IPM. These findings provide new evidence that MMPs and TIMPs could play a role in the turnover of IPM components.Cell homogenates and conditioned media from RPE isolated from mutant Royal College of Surgeons (RCS) rats with inherited retinal dystrophy had similar amounts of MMP-2 and TIMP-1 as those from congenic control rats. The secretion of MMP-2 and TIMP-1 from RPE cell cultures isolated from young and aged human donors varied widely. However, with increasing cell passage number, secretion of MMPs and TIMPs from human RPE increased dramatically. Also, growing human RPE on bovine corneal endothelial cell-generated extracellular matrix instead of plastic reduced the secretion of both MMPs and TIMPs. These data suggest that the integrity of Bruch's membrane may serve to regulate RPE functions in MMP and TIMP secretion and that extracellular matrices contain signals that regulate MMP and TIMP synthesis and/or secretion by the RPE.