应用同步法制备绒毛染色体标本的技术

本文根据绒毛在母体内悬浮于血窦中生长的生理特性,对186份绒毛标本进行了体外悬浮培养,并运用细胞同步化技术,建立了稳定可靠的绒毛染色体标本制备法。     

应用同步法制备绒毛染色体标本的技术

本文根据绒毛在母体内悬浮于血窦中生长的生理特性,对186份绒毛标本进行了体外悬浮培养,并运用细胞同步化技术,建立了稳定可靠的绒毛染色体标本制备法。     

外周血树突状细胞的体外培养

本文用塑料贴壁的外周血单个核细胞(PBMNCs)在含自体血浆、rhGM-CSF、rhIL-4和TNFα的RPMI1640培养基,37℃,5%CO2湿化空气培养树突状细胞(DCs).经10天培养,可获得大量具DCs形态学特征的细胞,其有很强的刺激同种淋巴细胞增殖功能,约30%的细胞表达HLA-DR和CD1a.健康志愿者每1×107PBMNCs可收获细胞约5×105.本文培养方法产率高,培养体系简单,用自体血浆代替小牛血清,培养过程简便,这些均适应临床治疗要求.     

体外培养细胞的加力实验装置

机械力对细胞生物学行为的影响是目前细胞生物学和细胞力学研究的一个重要课题。体外分离细胞和加载培养技术是细胞力学的基础之一和目前细胞力学研究的重要方法。大致可分为离心力加载、流体加载、单细胞加载、压力传导加载和基底形变加载技术。     

丙型肝炎病毒感染的体外培养细胞的抗原表达

建立丙型肝炎病毒（HCV）体外感染细胞模型,观察其感染细胞的HCV抗原表达,用抗HCV抗体和HCVRNA阳性及阴性血清感染MOLT-4细胞,制备细胞片进行免疫酶染色。结果显示HCV感染MOLT-4细胞4天和阴性对照HCV抗原均为阴性;感染后7天胞浆内可见HCV抗原阳性免疫反应产物;15天阳性细胞达到高峰,43天仍可见少量阳性细胞。结果表明HCV体外感染MOLT-4细胞胞浆内观察到HCV抗原表达。     

CIK细胞的体外培养与细胞毒作用

CIK（cytokine-induced killer）细胞在体外可以快速扩增,具有高效的非MHC限制性杀瘤活性,被认为是抗肿瘤过继细胞免疫治疗的新希望。随着细胞操作及基因修饰技术的日益精进,人们正逐渐改进CIK细胞的体外培养和处理方法,以便进一步提高CIK细胞的增殖率及特异杀伤力,使其更好地应用于临床。     

昆明白小鼠1细胞胚胎体外培养系统的研究

研究发现在有或者没有磷酸盐的条件下,葡萄糖均抑制昆明白小鼠ｌ－４细胞期胚胎的体外发 育。在不含葡萄糖和磷酸盐的ＨＥＣＭ－ｌ中,桑椹率为４０．０５％（７４／１６８）,而对照Ｇ－ＨＥＣＭ－１仅为 ８．１４％（７／８６）;不含葡萄糖含有磷酸盐的ＣＺＢ中,桑椹率为６７．１１％（９３／１５２）,而对照ＴＡＬＰ仅 ６· ６７％（６／９０）。用不含葡萄糖而含有１． ０ｍｍｏ１／Ｌ谷氨酸肢和０． １１ｍｍｏｌ／Ｌ ＥＤＴＡ的ＣＺＢ液,与兔输 卵管上皮单层培养细胞（ＲＯＥＣ）协同培养小鼠１细胞胚,７３．３３％（１１０／１５０）胚胎发育至桑椹胚, 但没有观察到囊胚形成、用上述ＣＺＨＲＯＥＣ系统培养小鼠１细胞胚４８小时（３－４细胞）,再移入 ＴＣＭ１９９＋１０％ＦＣＳ＋ＲＯＥＣ系统,有７６．７４％（６７／８６）胚胎发育至桑椹胚,９６／小时后,４０．７０％ （３５／８６）发育至囊胚。     

用绒毛组织微核评价母亲因素对人胚的致突变性

对507对欲作人工流产的夫妇进行断面调查及绒毛组织微核的测定,以探讨父母吸烟、饮酒及采取避孕措施等因素对子代的致突变性。结果显示:微核的发生率与母亲年龄、孕次、孕龄、既往流产史等自身的因素无关;各种避孕措施及少量饮酒均不具有诱发绒毛微核细胞率增加的作用。然而,男方吸烟、女方被动吸烟其微核细胞率为0.7645±0.0561‰高于饮酒组(0.5667±0.2004‰)和非吸烟对照组(05522±0.0616‰),与对照组有显著性统计学意义(P<0.05)。男方配偶吸烟兼饮酒者其微核发生率最高(0.7944±0.0754‰),说明吸烟与饮酒对诱发绒毛微核的产生有协同作用的趋势。本文并对男方吸烟和/或女方被动吸烟可诱发胚胎绒毛组织细胞DNA和/或纺锤体装置的损伤,进行了探讨,提出应引起人们的高度重视,并建议利用人类绒毛组织微核测定法作为监测环境致突变因子对人类下一代影响的方法。     

普通型脆性位点的研究进展

脆性位点是DNA合成被部分抑制之后,在中期染色体上发生的位点特异的缺口或断裂区域,它也是基因组不稳定的区域。通常脆性位点在体细胞中是稳定的,但它们在许多癌细胞中经常发生缺失或重排。在已发现的80多种脆性位点基因中,研究最多的是FHIT和WWOX,它们与肿瘤的发生发展密切相关。简要综述了普通型脆性位点的研究进展及其与肿瘤的关系。     

猕猴、白眉长臂猿、人类染色体普通型脆性部位和G-带的比较研究

以BrdU、FdU、MTX诱导猕猴、白眉长臂猿和人类染色体普通型脆性部位的表达,并对染色体脆性部位和染色体进化的关系以及三种灵长类染色体的同源性进行了比较分析。结果表明,近缘动物染色体同源区内的脆性部位在进化上是保守的,可作为染色体具有共同起源的标志,结合G-带的比较,可以用以阐明近缘动物染色体的同源性和染色体进化。     

一个小麦—黑麦染色体代换的细胞学鉴定

对从六倍体小黑麦与普通小麦的杂种后代中获得的矮秆抗病选系84056-1-36-1进行体细胞C-分带鉴定,结果表明,它的21对染色体中,有1对短臂带型与1R相似的黑麦染色体代换了小麦的1D。观察“中国春”双端二体1A、1B与该选系杂种F_1的花粉母细胞染色体配对,发现分别有82.56%和65.71%的细胞出现异型三价体,所有细胞至少有两个形态不同的单价体;而在“中国春”端二体IDL与84056-1-36-1的杂种中,端体不配对的花粉母细胞占100%,经C-分带后,另外1条单价体显示明显的端带。从上述这些结果推断84056-1-36-1为1R(1D)代换系。     

脆性X综合征的分子遗传学研究进展

脆性X综合征是常见的遗传性智力低下性疾病,其发病率高,临床表现复杂,遗传规律独特,对脆性X 综合征的发病机理和脆性X综合征筛查与诊断方法等方面的一些研究进展进行了综述.     

Murine Neuroblastoma Cells Express Ganglioside Binding Sites on Their Cell Surface

The ability of S20Y cholinergic, and N115 adrenergic, murine neuroblastoma cells to adhere to immobilized gangliosides was studied. Viable S20Y cells adhered more strongly to GM1-coated plastic wells than to those coated with GM2, GD1a, or GT1b. The oligosaccharide portion of GM1 inhibited adherence of S20Y cells to GM1-coated wells, indicating that the carbohydrate moiety of GM1 bore the recognition site. Analysis of S20Y cell adherence to wells coated with derivatives of GM1 indicated that the cells did not adhere to asialo-GM1 and adherence to the methyl ester or de-N-acetyl derivatives was significantly reduced. Expression of the GM1 binding sites by S20Y cells appears to be density dependent; cells harvested at the confluent stage of growth were more adherent than those harvested at the preconfluent stage. Trypsin treatment of the S20Y and N115 cells resulted in a loss of binding to GM1-coated wells, suggesting that the cell surface GM1 binding site is a protein. In contrast, N115 cells showed no significant difference in their adherence to wells coated with GM1, GD1a, GT1b, Gal-Cer, asialo-GM1, or the methyl ester of GM1 when assayed under the same conditions as those imposed on the S20Y cells. The N115 cells did show a reduction in adherence to GM2-coated wells, suggesting that they recognized the terminal galactosyl moiety.     

Subcellular Localization of Fragile X Mental Retardation Protein with the I304N Mutation in the RNA-Binding Domain in Cultured Hippocampal Neurons

1. Fragile X syndrome, the most common form of inherited mental retardation,iscaused by the lack or dysfunction of fragile X mental retardationprotein (FMRP). The I304N mutation in the RNA-binding domain of FMRP results in an exceptionally severe form of mental retardation.2. We have investigated the subcellular localization of FMRP and its I304N-mutated form in cultured hippocampal neurons and PC12 cells, using immunofluorescence microscopy. In PC12 cells, FMRP was predominantly localized to the cytoplasm and also to the processes after differentiation by NGF.3. In cultured hippocampal neurons, granular labeling was detected along the neuronal processes.4. Double-labeling with synaptophysin antibody revealed FMRP at synaptic sites in neurons.5. The I304N mutation did not appear to affect the transport of FMRP to dendrites or its localization at synaptic sites. Thus, FMRP is a synaptic protein and the severe phenotype observed in the patient with the I304N mutation is not produced by alterations in dendritic transport.     

Characterization of Somatotropin- and Prolactin-Binding Sites on Bovine Granulosa Cells Using Homologous Hormones

Specific binding of bovine somatotropin (BST) and bovine prolactin (BPRL) to cow granulosa cells from antrumcontaining follicles of different diameter was studied. Scatchard analysis of the data revealed a single type of low affinity BST-binding sites on the granulosa cells with dissociation constants similar to those for the BPRL-binding sites. The number of BST-binding sites on the cells decreased with increasing follicle diameter from 3-5 to 6-10 mm. However, the binding capacity to BPRL decreased only in the case of cells from follicles 11-20 mm in diameter. The findings are discussed in relation to the homologous binding phenomenon.     

蛋白编码基因中四倍简并位点碱基转换与颠换的规律研究

以同源性在７０％以上的６组同源蛋白为材料,从系统发生的角度研究发生在四倍简并位点上的转换和颠换的关系。考虑碱基组成对碱基替换的影响后,转换对颠换的优势在四倍简并位点也存在,但不如线粒体ＤＮＡ中显著。比较不同转换或颠换之间的关系发现,不同转换或颠换以十分接近的比率发生,Ａ－Ｇ转换与Ｃ－Ｔ转换的比率为０．９９：１,各种颠换相对于Ｔ－Ｃ转换的比率在０．６５－０．７３之间。进一步讨论了转换对颠换存在优势的原因,推测它可能与体内存在的诱变剂有关。     

鸡胚三类组织体外培养细胞的生物学特性分析

采用组织培养的方法获取鸡胚不同组织细胞,利用M199培养基进行原代、传代培养,经形态学观察、生长曲线绘制、分裂指数测定等进行生物学特性分析。实验表明,鸡胚不同组织细胞具有不同的生物学特性,从形态结构到生长周期都有明显差异。获得的躯体来源细胞、心来源细胞为成纤维型,肺来源细胞为上皮型;其中,躯体来源细胞生长能力最强,心来源细胞次之,肺来源细胞最慢,躯体来源细胞倍增时间最短;核型分析表明,该实验体外培养的鸡胚细胞染色体数目为78条。上述结果可为今后鸡胚不同组织细胞的深入研究提供实验材料和依据。