Sister chromatid exchange and DNA damage in human lymphocytes induced by air-dust in Lanzhou City: involvement in free radicals

The air-dust samples collected from petro-chemical industrial region in the suburb of Lanzhou and from a certain rural region 64 km away from the city were extracted, with a mixed solvent (benzene: hexane: isopropanol=7:2:1) for 8 hours. A strong free radical signal at g= 2.00 of air-dust itself and a hyperfine splitting EPR signal of extract from air-dust have been detected. The sister chromatid exchange frequency (SCE) was increased by extracts of both dusts from the industrial region and from the rural region. If a chemical is able to increase SCE up to twice as high as the control, this chemical is considered to be mutagenic and/or carcinogenic. The double SCE frequency concentration is 23 μg/ml for the dust extract obtained from the industrial region and 47μg/ml for that from the rural region. Extracts were able to damage to DNA template. Results indicated that the mutagenicity and/or carcinogenicity of the extracts obtained from the petro-chemical industrial region were stronger than that of the     

Chromosome mosaicism and maternal cell contamination in chorionic villi

While chorionic villus sampling allows both early and rapid prenatal diagnosis of chromosome disorders, the accuracy of this technique has not been fully established. Maternal cell contamination and pseudomosaicism represent two major sources of diagnostic error. Combined use of both direct chromosome preparations and villus cultures is important in overcoming these problems. Direct preparations of villus tissue allow recognition of maternal cell contamination of villus cultures. Conversely, villus cultures yield higher resolution chromosomes and may be helpful in differentiating between true versus pseudomosaicism when two or more cell lines are identified in direct chromosome preparations. Preliminary data suggest that analysis of direct preparations from multiple individually processed villus fragments may also be of value in this regard. Until more experience is gained, mid-trimester amniocentesis should be offered to CVS patients when mosaicism is encountered.     

A study of chromosomal aberrations in amniotic fluid cell cultures

This paper represents the analysis of 1916 routine amniotic fluid specimens harvested by an in situ fixation technique in a prospective study with regard to cultural chromosome anomalies. Excluding constitutional abnormalities, 2·9 per cent of 19432 cells analysed showed some form of chromosome anomaly, terminal deletions (57 percent) and chromatid/chromosome breaks and gaps (18 per cent) being the most frequent, followed by interchange aberrations (13 per cent) and trisomy (5 per cent). No case was found of more than one colony from the same culture showing the same anomaly without it being present in other cultures from the same fluid. The wholly abnormal colonies had a surplus of trisomies and from the mathematical considerations presented one may infer that these are likely to reflect the presence of abnormal cells in the amniotic fluid. Partly abnormal colonies appeared at a frequency that would correspond to virtual absence of selection against chromosomally abnormal cells when cultured in vitro. The aberrations found were similar to those seen as single cell anomalies, except for chromatid breaks and exchanges. The data suggest a basic preferential induction of trisomy for chromosomes 2,18,21, and the Y-chromosome. Structural aberrations showed a marked clustering of breakpoints around the centromeres. The frequency of mutant cells was low (1·4 × 10⁻³) before culture was initiated. At harvest, the frequency of abnormal cells was much higher (3 × 10⁻²) corresponding to 3 × 10⁻³ mutations per cell per generation accumulating over approximately ten generations in vitro.     

Chromosome banding in direct preparations of chorionic villi

Chorionic villus sampling (CVS) is now currently offered for first trimester prenatal diagnosis of genetic disorders. Chromosome analysis of CVS in direct and culture preparations is possible using modifications of standard banding techniques. We summarize our experience in applying QFQ, GTG, RBG, CBG, DA/DAPI, NOR, and SC differentiation protocols to direct preparations. Characteristic chromosome regions are properly labelled by these techniques, and analysis of 300 band stage karyotypes is consistently achievable on GTG banded direct preparations. However, banding of CVS direct chromosomes has proved to be difficult, and the analysis needs to be backed up by culture preparations.     

碘代乙酸体外暴露诱导斑马鱼(Danio rerio)淋巴细胞内源性凋亡可能机理研究

以斑马鱼(Danio rerio)为试验生物,经淋巴细胞体外暴露试验,从细胞内源性凋亡角度,研究了饮用水消毒副产物—碘代乙酸诱导斑马鱼淋巴细胞损伤的致毒机理.结果表明,在碘代乙酸的环境浓度1μg·L-1下暴露24、36、48和96 h后,细胞凋亡率从对照组的3.52%分别增加到15.89%、22.47%、40.76%和52.13%,与对照组差异均为极显著(p0.01);线粒体膜电位分别较对照组下降了32.9%、50.1%、68.6%和81.5%,与对照组差异均为极显著(p0.01);细胞色素C的相对释放量分别增加了0.85、1.37、1.86和2.66倍,与对照组差异均为极显著(p0.01).暴露36、48和96 h后,Caspase-3酶活性分别增加了0.49、0.86和1.43倍,与对照组差异均为极显著(p0.01);Caspase-9酶活性分别增加了0.73、1.41和1.88倍,与对照组差异均为极显著(p0.01);抑制凋亡Bcl-2基因的相对表达量分别下降27.0%、35.3%和52.3%,与对照组差异均为极显著(p0.01);而促进凋亡的Bax基因相对表达量分别增加1.1、2.3和3.2倍,与对照组差异均为极显著(p0.01).碘代乙酸体外诱导斑马鱼淋巴细胞凋亡的可能机制是,线粒体膜电位的崩溃导致细胞色素C持续从线粒体中释放,进而引起细胞凋亡下游通路的变化;Bcl-2基因和Bax基因明显参与了对凋亡的调控.此外,Caspase-9酶的活化导致Caspase-3酶活化,最终引起细胞凋亡.     

中国碳排放权交易市场发展历程 ——从试点到全国

碳排放权交易作为控制温室气体排放的一种市场化手段,相对于行政手段具有全社会减排成本较低、能够为企业减排提供灵活选择等优势。2011年以来,我国在北京、天津、上海、重庆、湖北、广东、深圳7个地方开展了碳排放权交易试点工作,为全国碳市场建设积累了宝贵经验。2017年年底,以发电行业为突破口的全国碳市场启动。2018年4月,国务院碳交易主管部门由国家发展和改革委转隶至生态环境部,生态环境部从推动碳交易立法、建立健全制度体系、加快基础设施建设、强化基础能力建设等方面稳步推进全国碳市场建设。本文梳理了我国碳市场的发展历程,并对全国碳市场的进一步完善提出建议。     

Oxidative stress and apoptosis of Carassius auratus lymphocytes induced by nonplanar （PCB153） and coplanar （PCB169） polychlorinated biphenyl congeners in vitro

Among all the 209 kinds of polychlorinated biphenyls (PCBs) congeners, nonplanar and coplanar PCB congeners have di erent levels of toxicity on mammal cells such as neuronal cells, but little is known about their toxicity on fish cells although PCB congeners usually have high bioaccumulation abilities in the detected fish bodies. This study showed that 2,2’,4,4’,5,5’-hexacholorbiphenyl (PCB153, nonplanar congener) and 3,3’,4,4’,5,5’-hexacholorbiphenyl (PCB169, coplanar congener) caused apoptosis on the isolated crucian carp (Carassius auratus) lymphocytes and the induced cytotoxicity was structure-dependent. According to the laser confocal microscope observations, apoptosis was clearly distinguished by condensation of nucleus, shrinkage and formation of apoptotic bodies. DNA fragmentation was detected by agarose gel electrophoresis. These typical morphological and biochemical characteristics indicate the occurrence of apoptosis on fish lymphocytes. According to the flow cytometry analysis, after the cells were exposed to 10 mol/L PCBs for 3 h, the apoptotic percentage induced by PCB153 was 23.41%, while that induced by PCB169 was even higher (31.03%). Furthermore, incubating PCBs with fish lymphocytes enhanced levels of reactive oxygen species (ROS) and malondialdehyde (MDA), clearly indicating the presence of oxidative stress and lipid peroxidation. Our data also demonstrate that the di erent cytotoxic e ects induced by coplanar and nonplanar PCBs were correlated with their structural characteristics and the coplanar congener was more cytotoxic than nonplanar congener. This study suggests that cytotoxicity mechanisms of the PCB congeners on fish lymphocytes depend on their planarity and chemical structures.     

A chromosome 21-specific cosmid cocktail for the detection of chromosome 21 aberrations in interphase nuclei

Fluorescent in situ hybridization (FISH) with a 21q11-specific probe (CB21c1) consisting of three non-overlapping cosmids has been applied to interphase amniocytes of pregnancies at increased risk for fetal aneuploidy (N = 78) and to interphase lymphocytes, cultured and uncultured, of patients referred for Down syndrome (N = 19 and 28, respectively). In the uncultured amniocytes, six chromosome aberrations were detected: three cases of trisomy 21, a triploidy, a de novo 46,XX,t(21q21q), and a mosaic 46,XY/47,XY,+dic(21)(q11)/48,XY,+dic(21)(q11), +del(21)(q11). In 15 cultured and 20 uncultured blood samples, FISH correctly diagnosed trisomy 21 (full or mosaic) at the interphase level, which was confirmed in all cases by subsequent karyotyping. Because of specific and strong signals in interphase nuclei, CB21c1 appears to be a useful tool for the rapid detection of chromosome 21 abnormalities.     

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.     

Rapid detection of numerical aberrations of chromosomes 13, 18 and 21 in chorionic mesenchymal cells

We have devised and evaluated a rapid screening method for the detection of numerical aberrations of chromosomes13, 18 and 21 in chorionic villus cells. We used non-radioactive in situ hybridization (ISH) with three chromosome-specific probes on overnight-attached mesenchymal cells from chorionic villi. A blind study was performed of 47 karyotypically normal samples, one triploid sample, two samples trisomic for chromosome 21, and two samples from a fetus with putative mosaicism (46/47, +21). All samples were hybridized with the chromosome 18- and 21-specific probes. Thirty samples were additionally hybridized with the chromosome 13-specific probe. The test could be completed within 3-4 days of sampling. In samples disomic with respect to the probed chromosomes, an average of 2 per cent (range 0-9 per cent) had three hybridization signals. By contrast, in the samples trisomic for the probed chromosome(s), 57 per cent (chromosome 13), 51 per cent (chromosome 18), and an average of 74 per cent (55-86 per cent) (chromosome 21) of the nuclei exhibited three signals. In the putative mosaic samples, the number of nuclei with three chromosome 21-specific signals ranged from 41 to 69 per cent. We conclude that this technique rapidly and clearly distinguishes between normal and trisomic/triploid samples, and consequently may be of use in future prenatal diagnosis.