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。     

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.     

硒对体外培养大鼠胚胎的发育毒性实验研究

本实验用全胚胎培养方法探讨了硒对体外培养大鼠胚胎的发育毒性，结果表明硒对胚胎的损害有明显的剂量—反应关系，硒浓度为2.0μg/ml，首先受累及的器官是视觉系统，提示该系统是硒的主要胚胎毒性作用部位，高浓度时，硒对反映胚胎生长发育和器官形态学分化的各指标均有抑制作用，6.0μg/ml剂量组胚胎畸形率为66.7%，主要表现是无眼泡，耳泡，心包积液，腋芽小，神经管未闭和体节异常，12.0μg/ml，死胎率高达88.9%。     

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。     

二氯苯醌对斑马鱼胚胎发育及超氧化物歧化酶的影响

2,6-二氯-1,4-苯醌（2,6-dichloro-1,4-benzoquinone,2,6-DCBQ）作为一种新型消毒副产物,因其高检出率及高含量的特点而备受人们关注.本研究以斑马鱼胚胎为实验材料探究了2,6-DCBQ的发育毒性.结果发现：斑马鱼胚胎在8~24 hpf（器官发育形成时期）对2,6-DCBQ暴露最为敏感,死亡率最高;2,6-DCBQ安全浓度为27.15 μg·L^-1,属于极度危险外源化学物.在本实验中,2,6-DCBQ对斑马鱼的生长没有显著影响,但高浓度（100和150 μg·L^-1）2,6-DCBQ会显著增加斑马鱼胚胎及仔鱼的畸形率.在实验过程中还发现,高浓度（≥ 80 μg·L^-1）2,6-DCBQ会明显抑制斑马鱼胚胎的心率.此外,≥ 30 μg·L^-1 2,6-DCBQ可提高超氧化物歧化酶（Superoxide dismutase,SOD）的活性,并影响Cu/ZnSOD和MnSOD的正常转录.本研究较全面地探究了2,6-DCBQ对斑马鱼早期发育阶段的发育毒性,可为2,6-DCBQ的毒性研究提供可靠证据.     

Relationship between metabolic enzyme activities and bioaccumulation kinetics of PAHs in zebrafish (Danio rerio)

Many studies have investigated bioaccumulation and metabolism of polycyclic aromatic hydrocarbons (PAHs) in aquatic organisms. However, lack of studies investigated both processes simultaneously, and the interaction between these two processes is less understood so far. This study investigated the bioaccumulation kinetics of PAHs and metabolic enzyme activities, including total cytochrome P450 (CYPs) and total superoxide dismutase (T-SOD), in zebrafish. Mature zebrafish was exposed to the mixture of phenanthrene and anthracene under constant concentration maintained by passive dosing systems for 16 days. The results showed that PAH concentrations in zebrafish experienced a peak value after exposure for 1.5 days, and then decreased gradually. The bioaccumulation equilibrium was achieved after exposure for 12 days. Both of the uptake rate constants (k_u) and the elimination rate constants (k_e) decreased after the peak value. The variation of PAH concentrations and metabolic enzyme activities in zebrafish had an interactive relationship. CYPs and T-SOD activities increased initially with the increase of PAH concentrations, but decreased to the lowest state when PAH concentrations reached the peak value. When the bioaccumulation equilibrium of PAHs was achieved, CYPs and T-SOD activities also reached the steady state. In general, CYPs and T-SOD activities were activated after exposure to PAHs. The decrease of PAH concentrations in zebrafish after the peak value may be attributed to the great drop of k_u and the variation of CYPs activities. This study suggests that an interactive relationship exists between bioaccumulation kinetics of PAHs and metabolic enzyme activities in aquatic organisms.     

水体中黑碳对斑马鱼胚胎的生长发育危害识别

黑碳(BC)是含碳物质不完全燃烧后产生的残留物,广泛分布于水体、土壤和大气中,目前对于BC对水生生物的生态效应研究还十分有限。通过燃烧煤油获得BC,以斑马鱼胚胎为模式生物,研究了质量浓度分别为0,0.05,0.5和5 mg/L的BC悬浮溶液对斑马鱼早期生命阶段的生长发育毒性。结果表明,煤油源BC对斑马鱼具有明显的毒性作用,与对照组相比,BC暴露明显增加了斑马鱼的死亡率和心率,显著降低了孵化率和体长,导致斑马鱼形态畸形并且畸形率呈浓度依赖性。5mg/L的BC对斑马鱼的各项生长发育指标影响并非是最大的,可能与BC颗粒在水溶液中的聚集有关。该研究为BC在水生环境中的危害识别提供实验依据。     

应用黑斑蛙胚胎测试化学品发育毒性的方法

选择合适的尤其是本土生物种进行化学品生态毒性评估,对于各国化学品的环境管理十分重要.本文选择我国本土两栖种黑斑蛙的胚胎为试验生物,以半数致死浓度LC50、半数致畸浓度TC50、致畸指数TI及最小抑制生长浓度MCIG为终点指标,建立了黑斑蛙胚胎发育毒性试验方法.以五氯酚、视磺酸、氯化镉、重铬酸钾为测试物研究了方法的敏感性,并以五氯酚为测试物研究了方法的可重复性.结果显示:五氯酚、视磺酸、氯化镉和重铬酸钾对黑斑蛙胚胎的LC50分别为572.3 μg·L-1、54.8 μg·L-1、6.8 mg·L-1和97.5 mg· L-1.五氯酚、视磺酸和氯化镉对黑斑蛙胚胎的TC50分别为246.5 μg·L-1、18.7 μg· L-1和3.4 mg·L-1,TI分别为2.3、2.9和2.0,MCIG分别为160.0 μg· L-1、10.1 μg·L-1和2.4 mg·L-1.重复性试验发现,五氯酚的LC50、TC50、TI及MCIG的变异系数分别为12.6％、18.0％、23.0％和18.6％.与文献中非洲爪蟾胚胎试验的数据比较,这些数据显示黑斑蛙胚胎与非洲爪蟾胚胎对测试物的敏感性存在一定的差异,各个终点指标的变异系数相当.因此,本文建立的黑斑蛙胚胎发育毒性试验可以用于化学品的发育毒性评价,为我国化学品环境管理提供技术支持.     

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...