哺乳动物细胞去核产物(胞质体及核体)的结构与功能研究

1967年Carter发现细胞松弛素可以诱发组织培养细胞的自发排核之后,Prescott(1972)借助细胞松弛素存在下的离心处理,使这一排核现象普遍化,从而确立了体外细胞去核的标准方法。经过不断改进(croce et al., 1974;Veomett et al., 1976;Lucas etal., 1976;Wigler et al., 1975),现在,这一技术已广泛应用于细胞学研究的各个重要领域(Mc Burney et al., 1979;Goldman et al., 1974;du Bols et al., 1980)。细胞去核技术及其应用的研究在我国已有初步开展(陈瑞铭等,1979;沈鼎武等,1980)。本实验对二种上皮型传代细胞系进行了去核手术,用扫描电镜和透射电镜对所获得的胞质体     

论Mufushania三叶虫分类

通过对我国己经正式发表的Mufushania三叶虫的18个种模式标本的头盖特征进行Q型聚类分析,并结合传统定性分析后,提出其中Mufushania shalangensis Zhang and Zhou inZhang et al.,1980,M.angustilimbata Zhang and Zhou inZhang et al.,1980,和M.kailiensis(Yuan in Yuan et al.,2002)(=Elrathiella kailiensis Zhou in Lu et al.,1974)等3种应从Mufushania属中删除,对余下15个种,通过修订,转移和归并为2个种:M.nankingensis Lin,1965和M.bella(Yuan in Yuan et al.,2002)。本文将对研究我国寒武纪第二世晚期褶颊虫类三叶虫系统演化、生物地理分区以及华北与华南生物地层划分和对比,提供重要依据。     

云南、贵州24种果蝇的核型研究

随着细胞遗传学的发展和染色体研究技术的改进,许多作者(Lin et al.,1974;Singh et al.,1979;Baimai,1980)应用染色体的显微照片对果蝇自然种组的有丝分裂中期染色体数目、形态等进行较细致的分析比较,使细胞遗传学分析对系统分类和物种形成赋予新的意义。     

核体合成自身DNA与病毒DNA的能力

近年来,用松胞素B(简称CB)诱导细胞去核的技术已被许多学者应用于细胞内大分子代谢及其调控过程的核质关系研究中(Colonne et al.,1980;Premveer et al.,1980;White et al., 1982)。我们在对CB诱导的去核产物的研究基础上(何大澄等1982),对核体内RNA的合成进行了研究,表明核体合成RNA的能力与核体贴壁和所带质膜的完整性以及胞质量多少有关(戎宪辉等1984)。此外,我们还研究了两种小RNA病毒和痘病毒的核酸在胞质体内的复制,证明它们的复制可以不依赖细胞核     

中国大鲵消化系统13种器官的蛋白水解酶种类和活性分析

蛋白水解对生命活动是必不可少的(Vassali et al., 1994),蛋白质的酶解修饰(Xu et al.,1999)、细胞的迁移、组织再生与修复、消化系统对食物中蛋白质的消化等均与蛋白水解酶有关(Baimbridge et al.,1992),许多病理过程也与蛋白水解酶功能失调有关(Teichert et al., 1989; Monard, 1988).因此开展大鲵消化系统各器官的蛋白水解酶种类和性质的研究,对了解大鲵消化系统各器官的功能、演化及大鲵的营养需求、食性、消化生理等是必要的.本文对大鲵消化系统各器官的蛋白水解酶特征进行了初步分析,现将结果报道如下.     

土耳其斯坦东毕吸虫的扫描电镜观察

血吸虫类中如血居科(sanguinicolidae)的Aporocotyle simplex Odhner,1900、裂体科(Schistosomatidae)的日本血吸虫(Schistosoma japonicum)曼氏血吸虫(S.mansoni)及埃及血吸虫(S.haematobium)等多种血吸虫均经扫描电镜观察(Johnson and Moriearts,1969;Silk et al., 1969;Robson and Erasmus, 1970; Miller et al., 1972; Kuntz et al.,1976、1977;Voge et al., 1978; Thulin, 1980;及何毅勋和马金鑫,1980等)。关于土耳其斯坦东毕吸虫[Orientobitharzia lurkestanica(skrjabin,1913)Dut et Srivastava,1955]的体表扫描电镜尚无报告,而只见有此虫种体壁及肠管的透射电镜观察的资料(Lavrov and Fedoseenke,1978)。本文部分作者最近在内蒙东部兴安岭以南部分地区进行牛羊土     

氨基酸、肽和蛋白质的色谱分析及色谱法制备

<正> CA 97(01)3041c J 势垒色谱:蛋白质分离的高压液相色谱法Ruckenstein,E,et al.,Sep.Sci.Technol.,1982,17(6),763—83, 英文CA 97(01)3046h J 运用精氨酸衍生物的亲和方法Ishii,Shinichi,et al.,Methods Enrymol.,1981,80(Proteolytic Enz- ymes,PT.C)842—8,英文     

人胃癌结合β半乳糖苷凝集素对不同细胞的凝集作用

本文使用去唾液酸胎球蛋白柱从人溃疡型胃癌中分离纯化出一种对β半乳糖苷键有高亲和力的凝集素(LBP)。乳糖是其有效抑制剂。研究结果表明,LBP既能促进正常细胞间的凝集,又能促进或介导人胃癌细胞与正常细胞间的粘附;共价结合LBP的琼脂糖颗粒对红细胞的吸附能力增强。而这些作用都可以被乳糖完全或部分抑制。结果提示,LBP可能是在人胃癌细胞定向转移中起作用的一种重要分子。     

真菌原生质体技术在遗传学和虫生真菌研究领域中的应用

原生质体融合技术自70年代中到80年代末的十几年间有着相当快速的发展过程,从无机试剂诱导的融合(Binding,1974;Ferenczy,1975)到PEG辅助融合(Kao,1974)以至发展到电场诱导原生质体融合(Zimmermann, 1980; Halfmannet al.1983; Call et al. 1984;Broda et al. 1987; Foerster et al.,1986; Fikus et al., 1985; Magae etal., 1986; 汪和睦1986a,b;张博润等,1986;彭卫宪等,1987)反映了原生质体融合方法的演变全貌。随着原生质体融合手段的不断更新,在遗传学研究领域中,利用这项技术进行     

Kit基因无义突变导致W-3Bao小鼠显性白斑形成

Kit(W)基因是一种原癌基因,在小鼠中该基因位于第5号染色体距着丝粒约42 cM处,其编码的蛋白质是具有酪氨酸激酶活性的干细胞生长因子受体,为酪氨酸激酶受体信号通路的跨膜分子。在人类及小鼠,kit及其配体kitl突变都可能引起不同程度的贫血、肥大细胞减少、毛色变白和生育能力下降或丧失等症状(Rajaraman et al.,2002;Broudy,1997;Rajaraman et al.,2003;Kapur et al.,1999),同源的kit基因突变在猪及禽类都表现为显性的白色斑点(邓素华等,2000)。在先前的研究中,本课题组通过ENU诱变获得6种白斑突变小鼠,通过连锁分析法以微卫星为连锁标…     

The appearance of 1,25-dihydroxyvitamin D3 receptor during chick embryo development

The appearance of the 1,25-dihydroxyvitamin D3 receptor in intestine, kidney, and chorioallantoic membrane of chick embryo was followed by sucrose density gradient sedimentation analysis and Scatchard plot analysis. The receptor from each of these organs sediments as a single 3.7S component. At 19 days of embryonic life, intestine had the highest specific 1,25-dihydroxyvitamin D3 binding activity followed by kidney and chorioallantoic membrane. The 1,25-dihydroxyvitamin D3 binding activity increased gradually at 12-15 days and rapidly until 20 days in intestine. In kidney, this protein increased rapidly from 12 to 16 days and did not change subsequently. In chorioallantoic membrane, the receptor increased slowly from 8 through 15 days, rapidly until 19 days, and decreased at 20 days. The injection of hydrocortisone into the chick embryo at 10 days increased receptor number in intestine, kidney, and chorioallantoic membrane by a factor of 2 at 12 days. Injection of this hormone after this time had little or no effect.     

Separation and Characterization of Four Forms of β-N-Acetylhexosaminidase from Chicken Brain

Chicken brain beta-N-acetylhexosaminidases from embryos (16 and 21 days old), newborns (1 and 4 days old), and adults (3 1/2 months and 2 years old) were separated into four different forms by ion exchange chromatography on diethylaminoethyl-cellulose. Three of these forms were "acid" hexosaminidases (I, IIA, and IIB), and the fourth was a "neutral" form. Throughout development of the chicken, forms IIA and III maintained the same activity ratio, whereas that for form I decreased and that for form IIB showed an increase.     

The role of GTP in the regulation of two forms of AMP deaminase from chicken kidney

1. Two molecular forms of AMP deaminase have been revealed by phosphocellulose column chromatography of the chicken kidney extract. 2. The chromatographic, kinetic and regulatory properties of these two forms were similar to these of two enzyme forms previously found in the chicken liver, lizard liver and in rat small intestine. 3. GTP exerted different effect from MgGTP on the activity and kinetic parameters of both AMP deaminase I and II from chicken kidney.     

A radiochemical assay for N-acetyl-L-aspartate amidohydrolase (EC 3.5.1.15) and its occurrence in the tissues of the chicken

A simple and rapid radiochemical method for the determination of N-acetyl-L-aspartic acid amidohydrolase (EC 3.5.1.15) activity using ion exchange chromatography has been developed. The activity of this enzyme in the developing brain and some non-nervous tissues of the chicken has been determined. No activity of the enzyme could be detected in the brains of chick embroys prior to 14 days of gestation; activities gradually increased thereafter to adult levels which are about 60% of that found in the adult rat. In non-nervous system tissues of the adult chicken, activities varied from high levels in the kidney to low levels in heart and breast muscle. Treatment of the homogenates of the adult tissues with a detergent significantly increased the enzyme activity, suggesting that a portion of the enzyme is membrane bound.     

Studies on L-arginase in developing rat small intestine, brain, and kidney. I. Ontogenic evolution of arginase isoenzymes

The adult patterns of arginase isoenzymes in rat intestine, kidney, and brain are nearly identical and consist of two forms, cationic A1 and anionic A4. In this paper, the organ-specific maturation of the enzyme equipment in these tissues is reported. The activity of arginase in all tissues studied could be detected on the 13th to 16th days of gestation. In fetal intestine and kidney the arginase activity is low, and persists up to the weaning time when the rapid, 10-fold rise of the enzyme activity occurs. However, the adult pattern of arginase isoenzymes in these tissues is accomplished in different ways. In the intestine, arginase A1 appears in fetal life and is the only form of the enzyme till the 19th to 21st days of postnatal life when the second form of arginase, A4, appears and rapidly accumulates, being exclusively responsible for the rise of the total enzyme activity at the time of weaning. In kidney, arginase A1 alone is present in the early fetal period. Arginase A4 appears 3-4 days before birth and its activity persists unchanged within the first 2 weeks of postnatal life. The intensive rise in total specific activity of kidney arginase at weaning is due to the accumulation of preexisting arginase A4. In brain, the adult pattern of arginase isoenzymes is achieved earlier than in other tissues. Both forms, A1 and A4, occur on Days 13-14 of gestation.     

AN ANALYSIS OF THE EFFECT OF "CONDITIONED MEDIUM" UPON THE CELL CULTURE AT LOW DENSITY

A favorable effect of “conditioned medium” upon outgrowth of the cell culture with low density in vitro was analysed with the cells of chicken embryos. For preparing “conditioned medium”, cultures with a large number of cells were made with muscle, kidney, lung, liver and skin, while the biological activity of the medium was assayed by using the culture of a small number of the lung secondary cells. A use of “conditioned medium” was found to be necessary for encouraging the outgrowth of the cultured cells below a critical inoculum size. Of the various types of the media tested, the medium conditioned with muscle was most effective. “Conditioned medium” contained at least two different active factors, the first to enhance the plating efficiency of the inoculated cells to the surface of the culture dish, and the second to promote further outgrowth of the plated cells. “Conditioned medium” taken out of the mass culture at its exponentially growing phase had only the second factor, while that taken out of that at its stationary phase contained both factors. An activity of the first factor was not detected, when the mass culture was kept in such condition that the collagen synthesis was inhibited. The factor for enhancing the plating efficiency was eliminated from “conditioned medium” by preincubating the cells, before assaying the effect of the medium.     

池蝶蚌Sox2 基因在不同组织及不同月龄精巢中的表达

Sox 基因家族在胚胎发育过程和性别分化中起重要作用, 为研究池蝶蚌中Sox 基因的功能, 以人SRY基因HMG-box 保守区的序列设计简并引物, 以雌、雄池蝶蚌基因组DNA 和精巢cDNA 为模板进行扩增, 获得了2 个不完全相同的序列, 分别为DNA-HMG1、DNA-HMG2 和cDNA-HMG, 长度均为220 bp, 编码73个氨基酸。与人等物种Sox1、Sox2、Sox3 及Sox14 有很高的同源性, 雌雄个体之间没有序列差异性。采用RACE-PCR 扩增获得了池蝶蚌性腺Sox2 部分cDNA 片段, 长度为1774 bp, 该序列核苷酸与欧洲帽贝的SoxB和人类的Sox2 的同源性最高; 在部分开放阅读框249 个氨基酸残基中, 具有Sox 家族典型的HMG-box 结构域, 与人类、小鼠、原鸡和斑马鱼等Sox2 的HMG-box 同源性为98%。为了解该基因在各组织中的表达情况,采用实时荧光定量PCR 方法分析了外套膜、闭壳肌、鳃、肠、肝、肾、精巢和卵巢在内的8 种组织hs-Sox2的表达情况, 结果显示, hs-Sox2 基因在8 种组织中均有表达, 其中在肾脏中的表达量最高, 其次是肠与闭壳肌, 在雄性性腺中的表达量明显高于雌性性腺, 在肝脏中的表达量最低; 为了解hs-Sox2 在不同性腺发育时期的表达情况, 采用实时荧光定量PCR 方法分析了5 个不同月龄的精巢组织中hs-Sox2 的表达情况, 结果显示在39 月龄性腺的表达量最高, 其次是16 月龄性腺, 63 月龄蚌中的表达量最少。以上结果表明, hs-Sox2 基因可能参与了池蝶蚌精巢的发育及功能的维持。       

Tyrosine aminotransferase converting factor: kinetic properties, cellular localization, and tissue distribution.

Utilizing a more rapid procedure for determining tyrosine aminotransferase-converting factor activity, the kinetic properties of this factor were characterized further. Tyrosine aminotransferase-converting factor is a heat-labile substance present in the particulate fraction of rat liver that converts tyrosine aminotransferase form III to I at 4 degrees C. Analysis of the distribution of marker enzymes for mitochondria and lysosomes, and of converting factor, following differential and discontinuous sucrose gradient centrifugation indicated that this factor was associated with light lysosomes. The activity of converting factor was not altered following administration of cortisol. Converting factor activity, equivalent to that in liver, was also observed in particulate fractions from kidney and spleen, and to a lesser extent, in pancreas and salivary gland. No detectable activity was observed in brain, heart, small intestine, skeletal muscle, red blood cells, serum, or plasma. The presence of converting factor activity in kidney and spleen suggests that other proteins are substrates for this factor since tyrosine aminotransferase is virtually absent from these tissues. Alternatively, the absence of converting factor from other tissues need not indicate they are devoid of converting factor-like activity merely, that such activity in these tissues has different specificities and does not utilize tyrosine aminotransferase as a substrate.     

Induction of cytochrome P-450 RNA by porphyrogenic agents: on the nature of the coordinate induction with delta-aminolevulinate synthase in tissues of the chicken embryo.

1. We determined by cDNA-RNA solution hybridization analyses that in ovo administration of allylisopropylacetamide in combination with diethyl 1,4-dihydro-2,4,6-trimethyl-3,5-pyridinedicarboxylate increased the concentrations of cytochrome P-450 RNA in liver, kidney, and intestine of 18-day-old chicken embryos. 2. Similarly, the administration of testosterone to embryos caused elevations in the cytochrome P-450 RNA levels in liver and kidney. 3. The increases in cytochrome P-450 RNA concentrations occurred only in those tissues where elevations in delta-aminolevulinate (ALA) synthase activity and mRNA content were measured (liver, kidney and intestine) but not in tissues where the activity and RNA levels of ALA synthase did not change (heart, brain, lung). 4. The increases in the concentrations of the cytochrome P-450 RNA were not affected by loading embryos with ALA and FeCl3 at the time of administration of the inducers.     

Tyrosine aminotransferase converting factor kinetic properties,cellular localization,and tissue distribution

Utilizing a more rapid procedure for determining tyrosine aminotransferase-converting factor activity, the kinetic properties of this factor were characterized further. Tyrosine aminotransferase-converting factor is a heat-labile substance present in the particulate fraction of rat liver converts tyrosine aminotransferase form III to 4°C. Analysis of the distribution of marker enzymes for mitochondria and lysosomes, and of converting factor, following differential and discontinuous sucrose gradient centrifugation indicated that this factor was associated with light lysosomes. The activity of converting factor was not altered following administration of cortisol.Converting factor activity, equivalent to that in liver, was also observed in particulate fractions from kidney and spleen, and to a lesser extent, in pancreas and salivary gland. No detectable activity was observed in brain, heart, small intestine, skeletal muscle, red blood cells, serum, or plasma. The presence of converting factor activity in kidney and spleen suggests that other proteins are substrates for this factor since tyrosine aminotransferase is virtually absent from these tissues. Alternatively, the absence of converting factor from other tissues need not indicate they are devoid of converting factor-like activity merely, that such activity in these tissues has different specificities and does not utilize tyrosine aminotransferase as a substrate.