大鼠非致瘤四倍体细胞系的建立及细胞遗传学和酶学特征

从大鼠自然诱发的肉瘤细胞中,我们建立了一个四倍体细胞系(4n=84),命名为RC(ratcell)。它具有典型的成纤维细胞外形,能在玻璃表面贴壁生长,但不能生长在琼脂半固体培养基中。该细胞在含15％小牛血清的RPMI 1640培养基中生长良好,至今已连续繁殖112世代,细胞群体倍增时间约为15小时。染色体G-带分析表明,RC为整四倍体细胞,它的1条X染色体在第32至34区为均染区。RC细胞核仁组织者(NORs)活性显然比大鼠二倍体细胞NORs活性的加倍还高(P<0.001)。这个具有非常高NORs活性的RC细胞系对于研究细胞18S+28S rRNA基因转录活性的调控、基因表达与基因剂量关系有一定的意义。RC细胞还有异常高的磷酸酯酶活性,而且它的同工酶谱也与大鼠肌肉细胞明显不同。体内接种实验和扫描电镜的观察表明,RC是非致瘤细胞。RC细胞各号染色体的C-带图样与大鼠二倍体细胞无明显的差异。     

Influence of both coculture and BrdU on NOR activity of mouse,rat and human cells

Summary The coculture of mouse PG19 cells with human MGC cells can significantly suppress nucleolar organizer region (NORs) activity of both PG19 and MGC cells. 5-bormodeoxyuridine (BrdU) can also significantly suppress the NOR activity of rat RC cells, human MGC and Hela cells, and mouse PG19 cells: i.e. the average number of Ag-NORs and the number of chromosomes bearing Ag-NORs per cell decrease significantly. The degree of the suppression increases with increase in both BrdU concentration in the culture medium and BrdU treatment time. The suppressed NOR activity of the PG19 cells can gradually be restored when the BrdU-treated cells are transferred into BrdU-free medium for 50 h. In PG19 cells deoxycytidine (dC) can reverse the suppression of NOR activity caused by BrdU. Coculture plus BrdU treatment suppress the NOR activity of PG19 cells more severely than BrdU treatment alone. In coculture medium containing 30 g BrdU/ml, dC can also reverse the suppression of the NOR activity of PG19 cells but not that of the MGC cells. The degree of the reversion in the coculture plus BrdU treatment is significantly lower than that found with BrdU-treatment alone.     

The effect of 1-β-d-arabinofuranosyl-cytosine on the expression of the common fragile site at 3p14

Summary The effect of the G₂-treatment of 1--d-arabino-furanosyl-cytosine (araC) on the expression of the common fragile site at 3p14 (FRA3B) was studied. A significantly increased frequency of FRA3B induced by G₂ treatment of araC was found in the lymphocytes grown in folate-deficient medium (positive rate 100%). A relatively low frequency of FRA3B was also induced in the cultures with folate in four of the seven subjects. These is a synergistic effect between araC and growth in folate-deficient medium on the induction of FRA3B. The results suggest that the DNA lesions related to the expression of FRA3B induce the long-patch repair and that the low DNA polymerase activity and inefficient repair process during G₂ phase is involved in the expression of FRA3B.     

Clastogen-induced chromosomal breakage as a marker for first trimester prenatal diagnosis of Fanconi anemia

Summary Using cultured trophoblast cells obtained by chorionic villus biopsy, we diagnosed Fanconi anemia (FA) in two pregnancies and excluded it in eight pregnancies at risk for the syndrome. Baseline chromosomal breakage and breakage induced by diepoxybutane (DEB) were analyzed. Increased breakage was used as a marker for the syndrome. Our results were unambiguous and provide a reliable method for prenatal detection of FA in the first trimester of pregnancy.     

Changes in the biological activity of agropyrene on combined application with humus substances in the cultivation ofScenedesmus obliquus (TURP.) krüger

The work is concerned with the question of the effect of humic acids on the biological action of agropyrene, the effective substance from the rootstock of Agropyron repens (L.) P. BEAUV. The test organism was the alga Scenedesmus obliquus (TURP.) KRÜGER, cultivated in mineral solution. The basic biological test was the determination of the number and size of the cells of this alga. It was found that the way in which agropyrene acts on combined application with humus acids depends on which humus fraction is used. It can display a synergic action if humic acid is used, since this substance probably facilitates the penetration of agropyrene into the cells. On the other hand, fulvic acid counteracts the activity of agropyrene and this most probably by mutual binding and blocking of active groups and links.     

脊髓灰质炎病毒中Ⅰ9株基因组克隆及其部分结构分析

以脊髓灰质炎病毒RNA为模板,反转录合成了长链ds-cDNA。将合成的脊灰病毒中I9株ds—cDNA片段重组到质粒pAT153上,获得了约占脊灰病毒中I9基因组95％以上的cDNA克隆。对克隆的中I9cDNA部分片段作了限制性内切酶图谱分析和部分核酸的序列分析,发现脊灰病毒中I9部分核酸顺序有所改变。     

抗菌免疫核糖核酸组份的分离及其活性测定

<正> 抗菌免疫核糖核酸(iRNA)已用于条件致病菌感染的临床治疗,并对其免疫活性做了全面的研究。现已证实iRNA能够诱导特异性抗体的产生和传递特异性的细胞免疫,并能诱生干扰素和白细胞间素Ⅰ、Ⅱ等淋巴因子和单核因子。但制备的iRNA是含有多种RNA种类的混合物,为明确各组份的免疫学功能,我们对iRNA进行了分离,并测定不同组份的免疫活性。     

生物素标记寡核苷酸探针探测扩增的病理性突变珠蛋白基因

用末端转移酶催化生物素核苷酸底物(Biotin-ll-dUTP)共价连接在合成的寡核苷酸3’羟基末端,从而合成了两种寡核苷酸探针(β~T_(41-42)及β~A_(41-42))。用它们分别与克隆化扩增的正常和突变的β—珠蛋白基因片段杂变。结果表明该探针都具有与~(32)P探针相似的特异性,其杂交的灵敏度为2—3pg(特异序列)。进而将探测HbS基因的正常和异常两种寡核苷酸19聚体(β~A_6和β~S_6)用~(32)P和生物素分别标记;将HbS杂合子病人的白细胞DNA经聚合酶链反应(PCR)法扩增,并以含正常β—珠蛋白基因的DNA片段作对照,与两种探针分别进行斑点杂交。所得结果完全一致;Hbs杂合子DNA对正常和异常探针都显出杂交信号,而正常DNA只与β~A探针显杂交信号。