大鼠肝和BERH—2肝癌5Kb BamHI DNA片段的克隆及其转录研究

本文克隆了大鼠肝和肝癌BERH-2 DNA的分子大小约为5kb的BamHI片段(Bam5族重复顺序)。从筛选出的克隆中取一个来自肝癌细胞基因组DNA的克隆片段H5 B-1和来自肝细胞基因组DNA克隆片段L5 B-4做进一步分析研究。采用RNA点杂交法对L5 B-4DNA片段的转录产物在细胞核、质间分布的研究结果表明:L5 B-4 DNA片段转录产物在肝癌细胞的总核RNA,poly A~ 核RNA和poly A~-核RNA中的相对量,比正常肝细胞的相应RNA组分低;在肝癌细胞的总胞质RNA和多聚核蛋白体RNA中的相对量,则比正常肝细胞的相应RNA组分高;其转录产物在poly A~ 核RNA中的相对含量高于其他细胞RNA组分,几乎不存在于rRNA中。当用大鼠肝和肝癌总RNA进行RNA点杂交比较时其转录产物在肝癌细胞中的相对含量则高于正常肝。结果提示,L5 B-4 DNA片段的转录产物从细胞核向细胞质内转运,肝癌细胞明显高于正常肝细胞。以H5 B-1 DNA片段代替L5 B-4 DNA片段进行RNA点杂交,得近似的杂交放射自显影图谱。     

胎肝细胞输注与全胚注射液治疗再生障碍性贫血的机理探讨

本文对胎肝细胞输注或全胚注射液治疗再生障碍性贫血的可能机理作了一些实验性探讨。研究结果表明: 1.胎肝细胞在培养或解体过程中释放某些刺激红系造血的因子,有利于已经损伤的造血功能的恢复。 2.对正常小鼠注射无细胞胎肝制剂或全胚注射液后,骨髓红系细胞的分裂指数明显升高,骨髓中粒/红比值趋于降低,反映了骨髓中红系细胞增生活跃的状态。 3.对正常小鼠注射无细胞胎肝制剂或全胚注射液后,外周血网织红细胞和腹腔巨噬细胞的吞噬指数趋于平行增高,其增高程度和持续时间随注射次数的增加而加强。 小鼠注射无细胞胎肝制剂或全胚注射液后,巨噬细胞吞噬指数的增加,反映了巨噬细胞激活,这种作用除了提高机体的非特异性免疫功能,增强机体的抵抗力外,还可能通过巨噬细胞的活化,直接或间接地调控机体红系细胞的增殖,因而,对巨噬细胞在造血调控中的作用以及它在再生障碍性贫血发病机理研究中的意义提出了讨论。     

不同发育年龄大鼠肝细胞及其溶酶体对急性低氧的应答

人工低压舱内模拟高原低氧24h,并与2300m对照组比较,观察不同发育年龄大鼠SGOT活力,肝溶酶体总酸性磷酸酶、非沉淀酸性磷酸酶和芳基硫酸酯酶活力及肝重、肝细胞糖原、蛋白和总脂含量的变化。在海拔5000m高度,10天鼠各酶活力、570天鼠总酸性磷酸酶和芳基硫酸酯酶活力明显升高;35和75天鼠各酶活力未见显著变化;在海拔8000m高度,各年龄组鼠上述各酶活力均显著升高。随着海拔高度的升高,各组大鼠肝重呈不同程度的下降,肝细胞糖原含量非常明显地减少,35和75天鼠8000m组全肝蛋白含量下降明显,10、35、75天鼠肝细胞总脂累积。上述结果综合分析表明:低氧致使大鼠肝细胞损伤属一普遍性效应,新生期和老年期大鼠肝细胞耐低氧能力不及幼年期和成年期大鼠。     

人肝癌细胞表皮生长因子受体以及佛波酯对它的调度

Using radioligand binding assay, the presence of epidermal growth factor (EGF) receptors in cells of two human liver cancer cell lines, BEL-7402 and SMMC-7721, was demonstrated. The ligand binding data were analyzed by a computer program. The dissociation constants (KD) of the ligand-receptor binding complex at equilibrium for 7402 and 7721 cells were 1.2 nM and 0.8 nM respectively, and their number of EGF receptors per cell were 6.2 x 10(4) and 2.5 x 10(4) respectively. After the treatment of cells with phorbol 12-myristate 13-acetate (PMA), no change either in the affinity or in the number of EGF receptors was found in 7721 cells. However, in the case of 7402 cells, while the number of receptors, like 7721 cells, remained unchanged, the affinity of EGF receptors displayed a time dependent modulation after PMA treatment. It dropped within the first hour to a KD value of 3.0 nM and then gradually returned to the normal control value at 48 hours or even slightly higher than normal (0.95 nM) at 96 hours of treatment. The modulation or down-regulation of EGF receptors by PMA in 7402 cells was paralleled by the simultaneous inhibition of DNA synthesis in these cells as evidenced from their reduction of 3H-TdR uptake. It is not clear what is the basis for the differences found between 7402 cells and 7721 cells in their number of EGF receptors per cell and their responsiveness to PMA treatment. It might be related to their difference in autocrine secretion of alpha-transforming growth factors.(ABSTRACT TRUNCATED AT 250 WORDS)     

组胺H2受体拮抗剂对骨髓造血重建的影响

用组胺H_2受体拮抗剂(甲氰咪胍或呋喃硝胺)处理正常和亚致死量γ-射线照射小鼠,探讨正常体内造血和再生骨髓中造血重建与组胺受体的关系。发现非毒性剂量的甲氰咪胍对正常小鼠骨髓多能造血干细胞(CFU-s)无抑制作用,但可抑制小鼠体内粒单系祖细胞(CFU-GM)的生长和亚致死量照射后CFU-s产率的恢复。组胺可能与骨髓的再生有关,组胺H_2受体拮抗剂可抑制骨髓的造血重建。     

氨甲酰胆碱和Impromidine对CFU—S细胞周期的影响

正常情况下处于S期的CFU-S比例低于10％。氨甲酰胆碱(Cach 10~(-13)—10~(-9)mol/L)和Impromidine(Impro 10~(-9)—10~(-4)mol/L)在体外与小鼠骨髓细胞短时培育后,增加了CFU-S对细胞毒剂羟基脲的敏感性。反应最大时,9d和13dCFU-S的减少率分别是32.8和60.6％(Cach)以及38.4和49.5％(Impro)。这种效应可分别被胆碱能N受体阻断剂筒箭毒(10~(-6)mol/L)和组胺H_2受体阻断剂甲氰咪呱(10~(-6)mol/L)所阻断,表明9d和13d CFU-S表面胆碱能N受体和组胺H_2受体的密度或活性存在差别,再次证实了CFU-S是一个不均一的细胞群。