基于代谢组学技术分析磷酸三苯酯诱导斑马鱼胚胎发育毒性的分子机制

磷酸三苯酯(TPhP)是广泛存在于环境介质和生物体内的一种典型有机磷阻燃剂。为探求TPhP诱发水生动物发育毒性的分子机制,本研究以斑马鱼为模式动物,将发育至2.5 hpf (hours post fertilization)的斑马鱼胚胎暴露于0.0025、0.1、1、10、100和1 000μg·L~(-1)TPhP溶液至7 dpf (days post fertilization),考察斑马鱼胚胎生长发育指标和线粒体功能的变化,通过代谢组学分析揭示相关分子机制。结果表明,环境相关浓度(0.0025、0.1和1μg·L~(-1)) TPhP对斑马鱼胚胎发育无显著影响,但是轻微干扰了斑马鱼的代谢过程。100和1 000μg·L~(-1)TPhP暴露引起斑马鱼心跳速率、孵化率和线粒体膜电位明显下调,畸形率分别增加6.8倍和12.5倍,死亡率分别增加7.2倍和16.5倍。代谢组学分析发现,10、100和1 000μg·L~(-1)TPhP显著抑制斑马鱼氨基酸代谢,降低缬氨酸、亮氨酸和异亮氨酸水平,抑制氨酰-tRNA生物合成过程;同时引起葡萄糖糖酵解过程和三羧酸循环发生障碍。氨基酸和糖代谢异常可能是TPhP引起斑马鱼发育畸形的主要原因,线粒体功能紊乱可能是TPhP诱发三羧酸循环障碍的原因。上述研究结果为TPhP发育毒性机制分析提供了新思路。     

4种农药复合污染对斑马鱼仔鱼联合毒性效应

为探明农药混合污染对斑马鱼的联合毒性效应,以斑马鱼仔鱼为研究对象,开展了氯氰菊酯、咪鲜胺、马拉硫磷和杀螟硫磷等4种农药的联合毒性效应研究。研究表明,氯氰菊酯、马拉硫磷,杀螟硫磷和咪鲜胺对斑马鱼仔鱼的96 h-LC_(50)值分别为0.12、17.88、12.39和1.45 mg·L~(-1)。根据96 h-LC_(50)值采用等毒比(1∶1)进行二元及多元联合毒性试验。二元农药混合污染(氯氰菊酯+马拉硫磷、氯氰菊酯+杀螟硫磷、氯氰菊酯+咪鲜胺和杀螟硫磷+咪鲜胺)对斑马鱼仔鱼联合作用表现协同作用。马拉硫磷+杀螟硫磷对斑马鱼仔鱼联合毒性在24 h时表现为协同作用,在其他不同时间均表现为相加作用。马拉硫磷+咪鲜胺二元农药对斑马鱼仔鱼联合毒性表现为拮抗作用。氯氰菊酯、咪鲜胺、马拉硫磷和杀螟硫磷4种农药的所有三元和四元混合污染对仔鱼联合毒性作用均表现为协同作用。研究表明,在真实的环境中,农药以混合物形式存在可能增加其对水生生物的毒性效应,给生态环境造成严重影响。     

微塑料对斑马鱼胚胎孵化影响及其在幼鱼肠道中的积累

微塑料在环境中的广泛存在引起了研究者对其潜在的生物影响的关注.为评价微塑料对水生生物的影响,本文以模式生物斑马鱼胚胎及其幼鱼为研究对象,探究了两种粒径不同的微塑料：粒径0.5 μm的红色荧光标记聚苯乙烯微塑料（0.5RF-PM）和粒径10 μm的绿色荧光标记聚苯乙烯微塑料（10GF-PM）对斑马鱼胚胎发育的影响及其在出生后5 d的斑马鱼幼鱼肠道中的积累.结果表明,10GF-PM对出生后3 d胚胎孵化没有影响,0.5RF-PM在500 mg·L^-1时对胚胎孵化抑制率达37%;10GF-PM溶液浓度为10、100、200和500 mg·L^-1时,出生后5 d幼鱼平均存活率分别为80%、54%、44%和41%;对应浓度0.5RF-PM暴露溶液中,幼鱼平均存活率分别为62%、37%、25%和12%.暴露在10、100和500 mg·L^-1的10GF-PM溶液中0.5 h,幼鱼肠胃道中微塑料荧光值分别是0.06、0.53和1.84,对应浓度0.5RF-PM暴露溶液中,荧光值分别是0.63、2.32和3.45.将暴露的幼鱼转移至清水中,1 h内可观察到微塑料通过肠道排出体外,持续放置24 h后,对应GF-PM暴露幼鱼肠道内微塑料荧光值分别是0.03、0.08和0.56,对应RF-PM暴露幼鱼肠道内微塑料荧光值分别是0.06、0.41和1.56.微塑料对斑马鱼的影响与浓度和粒径有关：浓度越高,胚胎孵化率和幼鱼成活率越低;粒径越小,越容易在肠胃道内积累.     

四种重金属离子对河川沙塘鳢早期发育的毒性影响

以早期发育阶段河川沙塘鳢(包括胚胎和初孵仔鱼)为研究材料,研究Hg2+、Cr6+、Zn2+和Cu2+4种重金属离子对其的毒性影响.结果表明,4种重金属离子对河川沙塘鳢早期发育死亡率和孵化率影响大小依次为Hg2+ ＞Cu2+＞Zn2+ ＞Cr6+,沙塘鳢在神经胚期和出膜仔鱼期的死亡率较高,且在重金属离子处理初期,大多数低...     

Preimplantation genetic diagnosis at 20 years

First reported in 1990, PGD has evolved into a complementary form of prenatal diagnosis offering novel indications. DNA for PGD can be recovered with equal safety and facility from polar bodies I and II, blastomere (8 cell embryo) and trophectoderm (5–6 day blastocyst). Diagnostic accuracy is very high (>99%) for both chromosomal abnormalities and single gene disorders. Traditional application of FISH with chromosome specific probes for detecting aneuploidy and translocations may be replaced or complemented by array comparative genome hybridization (array CGH); biopsied embryos can now be cryopreserved (vitrification) while analysis proceeds in orderly fashion. PGD has been accomplished for over 200 different single gene disorders. Novel indications for PGD not readily applicable by traditional prenatal genetic diagnosis include avoiding clinical pregnancy termination, performing preconceptional diagnosis (polar body I), obtaining prenatal diagnosis without disclosure of prenatal genotype (nondisclosure), diagnosing adult-onset disorders particularly cancer, and identifying HLA compatible embryos suitable for recovering umbilical cord blood stem cells. Copyright © 2010 John Wiley & Sons, Ltd.     

石棉表面改性对石棉诱导人胚肺细胞产生羟基自由基的影响

为了研究用柠檬酸铝、混合稀土或亚硒酸钠预处理温石棉对石棉诱导人胚肺(HEL)细胞产生羟基自由基(·OH)的影响,笔者用不同浓度的柠檬酸铝、混合稀土或亚硒酸钠溶液浸泡温石棉1小时后,再将其与人胚肺细胞共同孵育,利用电子自旋捕获法测定反应体系中羟基自由基信号。结果显示,石棉可诱导人胚肺细胞产生羟基自由基,并且所产生的自由基信号(峰高)随石棉浓度升高而增强,呈一定的剂量依赖性。经3种化合物预处理的温石棉组人胚肺细胞所产生的自由基信号(峰高)均明显低于未处理温石棉组。上述结果表明,温石棉可诱导人胚肺细胞产生羟基自由基,但用3种化合物预处理温石棉,均可抑制温石棉诱导人胚肺细胞产生羟基自由基。     

几种化合物对温石棉诱导入胚肺细胞蛋白激酶C活性的影响

为了探讨几种化合物对温石棉诱导人胚肺（HEL）细胞蛋白激酶C（PKC）活性的影响，该研究用不同浓度的柠檬酸铝、混合稀土或亚硒酸钠溶液浸泡温石棉1小时后，再将其与HEL细胞共同孵育24小时，测定了HEL细胞胞浆和胞膜中PKC的活性。其结果显示：随着石棉剂量的增加，HEL细胞胞浆和胞膜中的PKC活性逐渐升高，均呈明显的剂量依赖性；经3种化合物预处理的石棉，其所诱导的HEL细胞胞浆和胞膜中的PKC活性均明显低于未处理温石棉组。笔者提示：温石棉可能通过改变细胞内PKC的活性进一步促进肿瘤的发生；用3种化合物预处理温石棉，均可抑制温石棉对HEL细胞PKC活性的诱导作用。     

Mosaicism of autosomes and sex chromosomes in morphologically normal,monospermic preimplantation human embryos

We have previously detected chromosome abnormalities in human embryos whilst identifying the sex for preimplantation diagnosis of X-linked disease. In this study we assess the incidence of these abnormalities, both for sex chromosomes and autosomes 1 and 17, using dual fluorescent in situ hybridization (FISH). Sixty-nine normally fertilized embryos of good morphology at the 6–10 cell stage (day 3 post-insemination) were examined. The embryos were spread whole using HCl and Tween 20 to dissolve the cytoplasm. Thirty-four embryos were analysed for the sex chromosomes and 35 for autosomes 1 and 17. All probes were directly labelled with fluorochromes allowing analysis in 2 h. Control lymphocytes demonstrated that the probes were of high specificity. For the sex chromosomes, five embryos were mosaic (15 per cent) with the remaining 29 being uniformly XX or XY. In no case was an XX nucleus found in an otherwise XY embryo, indicating that even though mosaicism for the sex chromosomes is present, such abnormalities would not lead to a misdiagnosis of sex. For the autosomes, 16 embryos were abnormal (46 per cent); one embryo was triploid, one was monosomic for chromosome 1, and ten others were diploid mosaics (three diploid/aneuploid, three diploid/polyploid, and four diploid/haploid). A further four embryos had variable chromosome numbers in the majority of nuclei which appeared to be the result of uncontrolled mitotic division. The presence of haploidy or double monosomy, which occurred in 15 per cent of nuclei, has important implications for the diagnosis of trisomies and dominant disorders.     

Chromosome analysis of multipronuclear human oocytes after in vitro fertilization

Multipronuclear human eggs are frequent after in vitro fertilization. Their chromosome analysis can provide useful information. Before cleavage it can confirm the suspected poly-ploidy. Among the cleaved multipronuclear eggs it provides an estimation of the incidence of the possible return to diploidy. Ninety-four multipronuclear eggs were fixed at the first, second, or third cleavage according to the air-drying method of Tarkowski with or without colchicine exposure: 60 were successfully analysed. Twelve were stopped before cleavage (six without colchicine treatment and six with colchicine treatment). They were polyploid, confirming the cytological observation. Forty-eight eggs cleaved and were stopped by colchicine treatment and karyotyped. Seventeen eggs (35 per cent) had produced diploid embryos. Mosaicism was frequent (15 cases, 31 per cent). Triploidy was not frequent (8 eggs, 17 per cent). Haploidy constituted the remaining cases (8 eggs, 17 per cent). Our data indicate that the initial count of pronuclei is a reliable test. Multipronuclear one-cell oocytes were confirmed to be polyploid. Furthermore, the developmental capacity of the multipronuclear oocytes is variable. Most of them cleaved. However, many multipronuclear oocytes led to diploid cleaving eggs.