肿瘤核仁形成区形态定量诊断的误差及规范化分析

本文采用包括自动图象分析技术在内的ＡｇＮＯＲ形态定量学方法,以大肠肿瘤为模型,进行了ＡｇＮＯＲ定量形态学研究的误差分析,以探讨肿瘤ＡｇＮＯＲ定量诊断规范化的可能。结果表明,染色条件、视场目标选择和细胞计数量是引起ＡｇＮＯＲ定量诊断的主要误差;恒定染色环境,正确选择欲测细胞及测定足够量的细胞是使ＡｇＮＯＲ形态定量诊断规范化的途径。     

正常造血细胞的核仁组成区计数定量研究

本文应用银染技术对正常造血细胞的核仁组成区进行了计数定量研究。结果显示,在粒系和红系中,随细胞的成熟,细胞中簇状ＡｇＮＯＲ的数量逐渐减少．而点状ＡｇＮＯＲ数量逐渐增多,无分裂能力的成熟细胞仅有少数点状ＡｇＮＯＲ。淋巴细胞中为一完全集结的银染颗粒,而巨核细胞内为各自分离的点状银染颗粒。本结果为正常造血细胞的核仁组成区提供了基础数值。     

维生素A酸诱导HL-60细胞分化后银染核仁形成区的研究

用Ag-NOR、Brachet反应和作者改进的悬浮培养细胞Ag-aNOR的TEM技术等,综合地对RA诱导HL-60细胞分化后银染核仁形成区的变化进行了研究。实验结果表明,银染NOR的均值在给药组与对照组细胞中差异不显著。对间期细胞进行Ag-aNOR反应,结果??表明其数量在给药组比对照组明显减少,同时发现Brachet反应显示富含rRNA的核仁数与Ag-aNOR数改变极为一致,提示虽然活性的rRNA基因的数量在RA诱导分化的HL-60细胞中不变,但其转录明显受抑,使rRNA合成减少,进而在细胞水平出现了恶性表达的核仁变得少而小、甚至消失的分化表型。HL-60细胞的Ag-aNOR电镜观察更进一步证明了这一结论。     

间期细胞银染活性核仁形成区的电镜观察方法

本文报告改进建立的透射电镜砚察间期细胞银染活性核仁形成区的方法简便易行,能在同一个细胞中从亚细胞和分子水平上清楚地显示银染蛋白、活性rDNA和rRNA三者间的关系。     

维吾尔,哈萨克及汉三民族rRNA基因活性及随体联合的比较研究

应用核仁组织区银染技术对维吾尔、哈萨克及汉族共90人的染色体银染核仁形成区(Ag-Nucleolar Organizing Region,Ag-NOR)频率和近端着丝粒染色体随体联合频率的研究发现:维吾尔、哈萨克群体Ag-NOR及随体联合的细胞数与次数均高于汉族群体(P＜0.01),具有显著性差异。维吾尔、哈萨克群体间上述的频率无显著性差异(P＞0.05),支持了两民族遗传上的相近性。这与民族起源和过去用民族学、人类学研究结果是一致的。     

水稻随体染色体的研究

分析了几种野生稻种及栽培稻品种随体染色体的数目,发现不同染色体组型的稻种之间随体染色体数目有一定差异,其中染色体组ＡＡ的有１～２对,随体染色体因品种或样本的来源而异,染色体组ＢＢ的有３对随体染色体,染色体组ＣＣ的有２对随体染色体,染色体组型为ＢＢＣＣ的Ｏｒｙｚａｍｉｎｕｔａ可能有４对随体染色体,而染色体组型为ＣＣＤＤ的Ｏ．ｌａｔｉｆｏｒｉａ则有２对随体染色体。在栽培稻中,不同品种之间随体染色体的数目也存在差异,一般而言,籼稻品种具有两对随体染色体,分别为第１０和第１２染色体,粳稻品种只有一时随体染色体,为第１０染色体。     

东北梅花鹿染色体核仁组成区的银染观察

已知哺乳动物中18S+28S核糖体基因(rDNA)位于特定染色体的核仁组成区(NOR_s),银染技术能使哺乳动物染色体上的核仁组成区着色。银染核仁组成区是一种很好的遗传学标记。近年来,银染技术已开始用于追溯家畜的品种起源(王子淑等,1982)。还发现患白血病的黑白花奶牛的Ag-NOR_s出现率比正常牛显著下降(郭爱朴等,1983)。一些哺乳动物的Ag-NOR_s,如狗(张锡然等,1982)、灵长类(曹彼梅等,1981)、毛冠鹿(张锡然,1983)等已有过报道。本文应用银染技术,对东北梅花鹿(Cervus nippon hortulorum)的体细胞染色体的核     

睾酮对大鼠再生肝细胞核仁组织区影响的初步研究

以银染核仁组织区(argymphil nucleolar organiting regions,AgNORs)阳性颗粒数为指标,结合放射免疫技术,研究了大鼠部分肝切除(partial hepatectomy,PH)后,睾酮对余留肝细胞核仁组织区的影响。结果显示,pH后0～24h,各种方式(假手术、皮下注射芝麻油或睾酮)处理组,AgNORs数都下降,随后持续升高。在PH后48h和72h,低剂量(0．5ms／kg体重)睾酮对AgNORs数影响最显著,都明显高于其它两个睾酮处理组;中等剂量(2．5ms／kg体重)和高剂量睾酮(5ms／kg体重)处理对AgNORs数的影响与芝麻油处理组相比都无显著差异,但均高于假手术组;假手术后72h的AgNORs恢复到术前水平。血清睾酮含量测定显示,芝麻油处理组PH后0～48h睾酮持续下降,48h后不再下降,48h和72h时的睾酮浓度显著低于假手术组;低剂量处理,血清睾酮浓度在PH后0～24h下降,然后持续上升,在48h、72h达到假手术组的2～3倍;中等剂量和高剂量处理组,血清睾酮在PH后0～72h持续升高,是假手术组的6～7倍。以上结果初步表明,睾酮对PH后肝细胞中AgNORs的影响有两种情况：(1)在血清中的浓度是生理水平的2～3倍时,起促进作用;(2)6～7倍于生理水平时,基本无作用。     

rRNA genes are not fully activated in mouse somatic cell nuclear transfer embryos

The well known and most important function of nucleoli is ribosome biogenesis. However, the nucleolus showed delayed development and malfunction in somatic cell nuclear transfer (NT) embryos. Previous studies indicated that nearly half rRNA genes (rDNA) in somatic cells were inactive and not transcribed. We compared the rDNA methylation level, active nucleolar organizer region (NORs) numbers, nucleolar proteins (upstream binding factor (UBF), nucleophosmin (B23)) distribution, and nucleolar-related gene expression in three different donor cells and NT embryos. The results showed embryonic stem cells (ESCs) had the most active NORs and lowest rDNA methylation level (7.66 and 6.76%), whereas mouse embryonic fibroblasts (MEFs) were the opposite (4.70 and 22.57%). After the donor cells were injected into enucleated MII oocytes, cumulus cells and MEFs nuclei lost B23 and UBF signals in 20 min, whereas in ESC-NT embryos, B23 and UBF signals could still be detected at 60 min post-NT. The embryos derived from ESCs, cumulus cells, and MEFs showed the same trend in active NORs numbers (7.19 versus 6.68 versus 5.77, p < 0.05) and rDNA methylation levels (6.36 versus 9.67% versus 15.52%) at the 4-cell stage as that in donor cells. However, the MEF-NT embryos displayed low rRNA synthesis/processing potential at morula stage and had an obvious decrease in blastocyst developmental rate. The results presented clear evidences that the rDNA reprogramming efficiency in NT embryos was determined by the rDNA activity in donor cells from which they derived.     

Mutagenic selectivity of carcinogenic nitroso compounds I. N-methyl-N-nitrosourethane

The mutagenicity of the carcinogen methylnitrosourethane (MNUr) was examined in Drosophila with a view to the determination of its activity on heterochromatic loci (especially rDNA) relative to those in the euchromatin. Assays were made of the yield of rDNA mutations (bobbed: bb) relative to other X-chromosome recessive lethals and visibles [X(l + v)] in the same male germ cells after treatment with different doses (1–10 mM) and at various stages in spermatogenesis.Dose dependence followed the same pattern for all genic loci and germ cell stages. In all instances, the regression of mutation frequency on injected molar dose was approximately linear, but could better be described by a quadratic dose curve. In contrast, the mutagenicity pattern during spermatogenesis varied according to the target genes. The response of the euchromatic loci reached a peak among the earlier germ cells (probably the spermatocytes), whereas that for the heterochromatic sites (including rDNA) was maximal in mature sperm.Mutagenic selectivity for rDNA with MNUr, as indicated by the percentage bb/X-mutations, was among the highest for the intrinsically reactive carcinogens (alkylating and arylating agents). This correlates with the strong carcinogenicity of MNUr and adds further support to the concept that rDNA mutations might well be a crucial step in cancer initiation.     

Chromosomes of three freshwater stingrays (Rajiformes Potamotrygonidae) from the Rio Negro basin, Amazon, Brazil

Potamotrygonidae is the representative family of South American freshwater elasmobranchs. It is a monophyletic group containing 20 species grouped into three genera. Three species belonging to two genera of this family were collected from the middle Negro River, Amazonas, Brazil, and studied cytogenetically: Paratrygon aiereba, Potamotrygon motoro and Potamotrygon orbignyi. Paratrygon aiereba presented 2n = 90 chromosomes and 4M+2SM+10ST+74A. Both species of Potamotrygon presented 2n = 66 chromosomes and differed in their chromosomal formulas: P. motoro had 18M+12SM+10ST+26A and P. orbignyi had 22M+10SM+8ST+26A. No sex heteromorphism was detected. The Fundamental Number (FN) was 106 for the three species. A system of multiple NORs was found in the three species, but with interspecific differences in terms of location and position of the active Ag-NORs sites. Paratrygon aiereba presented only four sites on the short arms of two chromosomal pairs, both in terminal regions. Potamotrygon motoro presented seven sites, on the long and short arms, all in terminal regions of non-homologous chromosomes; P. orbignyi presented eight sites on the long arms, all in terminal regions, of non-homologous chromosomes. The constitutive heterochromatin was in pericentromeric regions of all chromosomes, and no significant interspecific difference was found in relation to this marker.     

水杨酸(SA)对果实成熟的影响

水杨酸是一种天然抑制剂,对植物有多种多样的作用,新近研究报道,它是乙烯生物合成的一种新的抑制剂。本文用水杨酸处理番茄、苹果、梨等果实,储藏在室温下,不时检查ＳＡ对果实贮存保鲜的影响。其结果表明ＳＡ处理的果实,ＰＧ活性比对照低,而硬度大,抗病力强。番茄,苹果和梨的无病好果率比对照果提高１０％以上。上述结果证明,用ＳＡ处理绿熟番茄及梨和苹果,能有效保存果实新鲜,增强抗病力和延长货架的寿命。     

喜树愈伤组织的诱导及诱导子对愈伤组织中喜树碱含量的影响

通过组织培养的方法分别对喜树营养器官进行愈伤组织的诱导和继代,筛选红色、黄色和褐色3个不同的细胞系;运用HPLC法测定各愈伤组织中喜树碱的含量,并在培养基中外加诱导子,分析其对愈伤组织中喜树碱含量的影响。结果表明,经幼叶所诱导的愈伤组织中喜树碱含量相对高于其它,红色和黄色的细胞系在不同的外植体中其喜树碱含量均高于褐色的细胞系。但从整体看来,愈伤组织中喜树碱的含量远远低于原植物中的含量。外加诱导子对愈伤组织中喜树碱的含量有影响,浓度为0.1 mg·L^-1,1 mg·L^-1的水杨酸和茉莉酸甲酯处理的愈伤组织中喜树碱含量提高,0.1 mg·L^-1的水杨酸对喜树碱的积累有明显的促进作用,茉莉酸甲酯的影响不大;浓度为3 mg·L^-1,5 mg·L^-1的水杨酸处理的愈伤组织中,喜树碱含量降低;3 mg·L^-1的茉莉酸甲酯处理的愈伤组织中,喜树碱含量降低,5 mg·L^-1处理的愈伤组织中未检测到喜树碱。