贫血患者血清红细胞生成素浓度的测定及临床意义

用小鼠胎肝细胞体外血浆凝块培养红系集落(Erythroid colong formig unit inculturc,E-CFUc)方法,以红细胞生成素(Erythropoietin,EPO)850323为标准试剂,测定正常人、贫血病人血清EPO浓度。实验用妊娠13～15d小鼠胎肝细胞。血清均经透析处理,培养液中加量最大不超过10％。EPO(850323)在培养液中浓度为2.5～100mU/ml。血清EPO(mU/ml)测定结果:28例正常人为48.O±17.7,12例再生障碍性贫血病人为946～>10000,1例巨幼细胞性贫血病人为500,1例缺铁性贫血病人为400和18例慢性肾功能衰竭病人则为94.2±87.6。结果表明:贫血病因对血清EPO浓度有影响。     

右美托咪定调控Nrf2/HO-1通路对H2O2诱导心肌细胞氧化应激损伤的作用研究

目的 探讨右美托咪定调控核因子E2相关因子2(Nrf2)/血红素加氧酶1(HO-1)通路对过氧化氢(H₂O₂)诱导心肌细胞氧化应激损伤的作用。方法 体外培养大鼠H9C2心肌细胞,设置对照组、H₂O₂组、1μmol右美托咪定+H₂O₂组、5μmol右美托咪定+H₂O₂组、10μmol右美托咪定+H₂O₂组。CCK-8法检测各组H9C2细胞增殖情况;酶联免疫吸附试验(ELISA)检测各组H9C2细胞丙二醛(MDA)、超氧化物歧化酶(SOD)水平;实时荧光定量聚合酶链反应(q RT-PCR)检测各组H9C2细胞Nrf2、HO-1 mRNA相对表达量;Western blotting检测各组H9C2细胞Nrf2、HO-1蛋白相对表达量。结果 与对照组比较,H₂O₂组H9C2细胞存活率、SOD水平、Nrf2、HO-1 mRNA及蛋白相对...     

三七总皂甙对红系祖细胞增殖调控机理的研究

采用造血祖细胞体外培养和造血生长因子检测等实验血液学技术，研究三七总皂甙对小鼠红系祖细胞调控的生物学机理。结果表明：三七总皂甙对正常或骨髓抑制一贫血模型小鼠的红系祖细胞增殖有明显促进作用。三七总皂甙亦可提高阿糖胞苷所致祖细胞的“自杀”率；经三七总皂甙诱导制备的脾细胞，Ｌ细胞培养上清液和红系祖细胞的直殖具有较高刺激活性。     

Monoclonal antibodies against human granulocytes and myeloid differentiation antigens

Monoclonal antibodies (MCA) were obtained by immunizing BALB/c mice with 99% pure granulocytes from normal donors or with a whole leukocyte suspension obtained from a chronic myelogenous leukemia (CML) patient, and then fusing the mouse spleen cells with a 315–43 myeloma cell clone. Four MCA were selected and studied using ELISA, immunofluorescence, cytotoxicity assays, and FACS analysis. Antibodies 80H.1. 80H.3. and 80H.5 (from normals) and 81H.1 (from CML) detected antigens expressed on neutrophils. Antibodies 80H.1 and 80H.3 (lgG) also reacted with monocytes but not with other blood cell subsets. Antibodies 80H.5 and 81H.1 (lgM) were cytotoxic and reacted strongly with most of the cells of the neutrophil maturation sequence. i.e., myeloblasts, promyelocytes, myelocytes, and mature granulocytes. Antibodies 80H.5 and 81H.1 also inhibited BFU-GM and CFU-E. Antigens recognized by 80H.3. 80H.5, and 81H.1 were expressed both on a proportion of cells from HL.60, KG.1, ML.1, and K562 myeloid cell lines, and on a proportion of blast cells isolated from patients with acute myelogenous leukemia. They were not found on lymphoid cell lines or lymphoid leukemia cells. These MCA recognize either late differentiation antigens expressed on mature neutrophils and monocytes (80H.1 and 80H.3) or early differentiation antigens (80H.5 and 81H.1) specific to the granulocytic lineage. They may be useful for a better definition of those antigens specific to hematopoietic stem cells and their relationship with normal or neoplastic hematopoiesis.     

Expression of the Tn antigen on erythroid cells from a patient with Tn syndrome

Summary In order to examine expression of the Tn antigen on erythroid cells from a patient with Tn syndrome, we applied a selective two phase liquid culture system for human erythroid progenitors in peripheral blood. The cells were analyzed with flow cytometry employing an anti-Tn antibody and a lectin ofVicia villosa which recognizes only the Tn determinant. In the second phase, the Tn antigen was expressed on the cultured cells from the patient on day 3 and Tn-positive cells reached 62.7% on day 9. On the other hand, Tn-positive cells were not detected in the volunteer's cultured cells. When the patient's cells were co-cultured with the cells from a healthy voluteer, the percentage of Tn-positive cells was much lower than the expected value, suggesting that the normal cells suppressed the expression of Tn antigen on the patient's cells.     

新生大鼠大脑皮层少突胶质细胞/Ⅱ型星形胶质祖细胞原代培养及形态

目的:研究不同诱导条件对大鼠大脑皮层O2A祖细胞生长活性的影响。方法:采用两次恒温摇床振荡培养法,倒置显微镜结合免疫荧光染色法鉴定细胞种类并判断纯度,透射电镜观察细胞超微结构。结果:O2A祖细胞胞体呈圆形,常有单极或双极突起.形成克隆球,A285标记阳性,免疫荧光鉴定细胞纯度可达90%以上。O2A祖细胞具有双向分化的能力,在不同诱导条件下可分化为星形胶质细胞或少突胶质细胞。电镜观察O2A祖细胞呈圆形或椭圆形,核仁可见,胞质内细胞器少,核周见成束胞质丝。结论:选择合适的培养基对O2A祖细胞纯化培养及诱导分化极其重要。     

Extensive training in a maze task reduces neurogenesis in the adult rat dentate gyrus probably as a result of stress

It has recently been shown that hippocampal neurogenesis can be modulated either directly or indirectly by ascending cholinergic inputs from the basal forebrain. In the present work, we sought to address whether extended training in a spatial navigation task would affect hippocampal neurogenesis in the presence of a severe and selective cholinergic depletion. Young female rats received stereotaxic injections of the immunotoxin 192 IgG-saporin into the basal forebrain nuclei and/or the cerebellar cortex. Starting from 4 to 5 weeks post-lesion, and for the subsequent 2 weeks, the animals were trained on paradigms of reference and working memory in the water maze and received single daily i.p. injections of bromodeoxyuridine (BrdU) at the end of each testing session. In line with previous observations, a dramatic 80% decrease in neuron proliferation was seen in the dentate gyrus of lesioned animals, as compared to vehicle-injected or intact controls. Interestingly, however, rats subjected to maze training over 2 weeks, irrespective of their learning success, exhibited significantly fewer newborn neurons than matched controls with no maze exposure. Thus, at least for the type of task used here, which has previously been shown to impose a certain degree of stress, extended training and learning does not appear to affect proliferation in the dentate gyrus.     

植物血凝素(PHA)对小鼠骨髓粒—单祖细胞增殖的影响

用体外琼脂培养法研究了PHA对小鼠骨髓CFU-GM的影响,结果表明,小鼠经腹腔注射PHA后,对小鼠CFU-GM产率和自杀率的影响随PHA的剂量不同而异,20～40mg/kg的PHA能明显提高CFU-GM的产率,其作用时间维持3天,剂量大于50mg/kg,可抑制CFU-GM产率。PHA组自杀率较对照组明显提高,提示适量的PHA在体内可促进小鼠CFU-GM的增殖,使更多的CFU-GM由Go期进入S期。将PHA直接加入无CSF的培养体系中,未见集落生长。此项结果提示PHA可能通过间接机制而非直接作用于CFU-GM。     

视网膜祖细胞干细胞特性及移植入视网膜后的研究

目的：研究视网膜祖细胞的干细胞特性及移植入视网膜后的存活和迁移。方法：体外培养胎龄18d 大鼠的视网膜细胞,用 RT-PCR、细胞免疫荧光方法鉴定其增殖分化；成年 SD 大鼠腹腔注射 N-甲基-N-亚硝基脲形成视网膜感光细胞退化的动物模型,培养的视网膜祖细胞用 CM-Di(?) 标记后,移植入模型鼠的玻璃体腔。结果：视网膜祖细胞体外表达中间神经丝蛋白 nestin；表达 Flk-1、Pax6及 Notchl 的 mRNA；能掺入 BrdU；分化后表达视网膜各类细胞特异性蛋白；移植后在实验组大鼠视网膜能存活及迁移,而在对照组中仅聚集在玻璃体腔。结论：视网膜祖细胞具有干细胞特性,移植入受损伤视网膜后,能存活、整合及迁移。     

α6 integrins participate in pro-T cell homing to the thymus

During embryogenesis, colonization of the thymic rudiment by hemopoietic progenitor cells depends on the adhesion of these cells to the jugular endothelium. Previously, we showed that progenitor T cells (pro-T cells) interact with α₆ integrins present on vascular endothelium. Here, we demonstrate that anti-α₆ integrin antibodies reduced the number of thymocytes up to 80 % in a congenic mouse model for thymus colonization by pro-T cells. In organotypic thymus cultures, the anti-α₆ integrin antibodies did not influence T cell development and proliferation. From this, we conclude that α₆ integrin participates in thymus homing. During mouse thymus ontogeny, α₆ integrin mRNA and protein expression was found as early as day 10 of development; at day 11, perithymic endothelial cells were α₆ integrin positive. Two α₆ integrin mRNA exist which are produced by alternative exon usage. The longer form, α₆, integrin, predominates during early embryonic stages, while the shorter α_6A form was present later during development. Although α₆, integrins can be displayed by immature thymocytes, strongest expression was found on intra- and perithymic vascular endothelium. These data suggest that α₆ integrins are involved in the homing of pro-T cells to the developing thymus by mediating adhesion of pro-T cells to the vascular endothelium.