Trophoblast-like cells sorted from peripheral maternal blood using flow cytometry: A multiparametric study involving transmission electron microscopy and fetal dna amplification

Three monoclonal antibodies (MAbs) against trophoblast (GB17, GB21, and GB25) and flow cytometry were used to sort trophoblast-like cells (TLCs) from peripheral blood of pregnant women. Sorted TLCs were processed for electron microscopy and fetal DNA amplification of the Y-specific sequences from mothers carrying male fetuses. At the ultra-structural level, most of the nucleated cells had the morphology of leucocytes, suggesting maternal contaminants, and we did not find the characteristic features of the free inter-villous trophoblast cells. Nevertheless, polymerase chain reaction (PCR) analysis showed an amplification of Y-specific sequences in two out of three samples of sorted TLCs. These results suggest that besides the maternal leucocytes, sufficient trophoblast nucleated fetal cells can be obtained using cell enrichment by sorting. This sensitive method holds promise for non-invasive prenatal diagnosis of fetal sex and if sufficient Y(positive) nuclei are found, for the diagnosis of selected numerical chromosome abnormalities.     

Immediate prenatal diagnosis of Zellweger syndrome by direct measurement of very long chain fatty acids in chorionic villus cells

We report a gas chromatographic-mass spectrometric method which allows the very long chain fatty acids content of trophoblastic tissue to be directly measured in samples collected by biopsy between 8 and 11 weeks of gestation. This method has been successfully applied to the detection of fetal Zellweger syndrome in two pregnant women who had previously delivered affected infants. In one of them, increased concentrations of C26:0 (0.254 versus 0.108±0.035 μ/mg proteins) and C24:0 (1.32 versus 0.815±0.325 μ/mg proteins) in trophoblast indicated that the fetus had Zellweger syndrome, a diagnosis confirmed by pathological findings after abortion. In the second case, the pregnancy was allowed to proceed, on the basis of normal concentrations of very long chain fatty acids in trophoblastic tissue, and its outcome was actually a healthy newborn.     

Detection of fetal HLA-DQα sequences in maternal blood: A gender-independent technique of fetal cell identification

The objective of this study was to detect fetal HLA-DQα gene sequences in maternal blood. HLA-DQα genotypes of 70 pregnant women and their partners were determined for type A1. We specifically sought couples where the father, but not the mother, had genotype A1. In 12 women, maternal blood samples were flow-sorted. Candidate fetal cells were isolated and amplified by using PCR primers specific for a paternal HLA-DQα A1 allele. Fetal HLA-DQα A1 genotype was predicted from sorted cells; amniocytes or cheek swabs were used for confirmation. Six of twelve sorted samples had amplification products indicating the presence of the HLA-DQα A1 allele; 6/12 did not. Prediction of the fetal genotype was 100 per cent correct, as determined by subsequent amplification of amniocytes or cheek swabs. We conclude that paternally inherited uniquely fetal HLA-DQα gene sequences can be identified in maternal blood. This system permits the identification of fetal cells independent of fetal gender, and has the potential for non-invasive prenatal diagnosis of paternally inherited conditions.     

BPA暴露对本体和子代胰岛β细胞转录组差异对比分析

双酚A(BPA)作为一种典型的环境内分泌干扰物,与二型糖尿病和肥胖症等代谢类疾病密切相关.在这项研究中,旨在探讨BPA直接暴露以及围产期暴露对本体和子代胰岛β细胞影响的差异,从而实现对不同易感人群的精准预防与治疗.通过对GEO数据库中GSE126297和GSE82175两个数据集进行分析,共筛选出108个共有的差异基因...     

纳米氧化锌颗粒物通过氧化应激和离子通道失调诱导大鼠气管上皮细胞凋亡的机理

纳米氧化锌具有广泛的工业用途,其生态安全性受到广泛关注,针对纳米氧化锌诱导的呼吸道细胞毒性及其作用机理研究尚不广泛.本研究分别采用不同浓度和粒径(30 nm和90 nm)的氧化锌颗粒物处理大鼠气管上皮细胞(rat tracheal epithelial cells,RTE cells),暴露时间为12 h,通过检测细胞内锌元素含量,细胞增殖抑制率,细胞凋亡率,凋亡相关caspsae 3基因与蛋白相对表达量,细胞内金属硫蛋白活性,ROS和MDA含量、细胞内Ca~(2+)-ATP酶和Na~+/K~+-ATP酶活性来分析纳米氧化锌诱导细胞毒效应机理.在90 nm纳米氧化锌高浓度暴露时,其细胞内锌元素浓度为0.845μg·L~(-1),约为低浓度暴露组的4.7倍,是30 nm低浓度暴露组的9倍;纳米颗粒物诱导的细胞增殖和凋亡毒效应具有剂量和尺寸依赖效应;30 nm处理组的pro-caspase 3和cleaved-caspase 3蛋白表达量均高于90 nm暴露组;暴露浓度为10 mg·L~(-1)的90 nm处理组的金属硫蛋白增加量为0.533μg·L~(-1),增幅达到46%;不同粒径氧化锌颗粒物处理后,细胞内ROS和MDA含量显著上升,且30 nm处理组结果均高于90 nm处理组;纳米氧化锌颗粒物暴露诱导细胞Ca~(2+)-ATP酶和Na~+/K~+-ATP酶活性显著下降,30 nm氧化锌颗粒物暴露组,其Na~+/K~+-ATP酶活性分别是对照组的1.8倍和3.5倍.纳米氧化锌颗粒物进入RTE细胞,通过干扰锌在细胞内代谢,诱导细胞内ROS和MDA水平升高,产生氧化应激,进而诱导细胞凋亡是导致纳米氧化锌产生细胞毒性的主要原因之一.纳米氧化锌会导致细胞内Ca~(2+)-ATPase和Na~+/K~+-ATPase活性下降,离子通道失调,破坏细胞内离子平衡,进一步造成细胞凋亡.     

空气阴极微生物燃料电池的构型优化及其快速测定BOD的性能评价

设计了一种新型双室空气阴极微生物燃料电池(MFC)并将其作为生物传感器,与传统双室空气阴极MFC进行对比,考察其电化学性能及用于快速检测BOD的性能。结果表明：新型空气阴极MFC可有效提高功率密度并降低内阻,其功率密度最高为897 mW·m⁻²,而内阻最低为92 Ω;该MFC可用于直接快速检测高浓度有机物的BOD,对醋酸钠底物的线性检测限为1 280 mg·L⁻¹,在此底物浓度下MFC的检测时间为31.2~66 h,线性可决系数R²为0.97~0.99;对于GGA底物的线性检测限为1 250 mg·L⁻¹,在此底物浓度下MFC的检测时间为33~67 h,线性可决系数R²为0.98。本研究可为MFC型BOD检测传感器的性能优化提供参考。     

Mitochondrial toxicity and in vitro biological effects of ambient PM2.5 from an urban site in China

Abstract

The roles of PM_2.5-induced mitochondrial damage and oxidative stress on mast cell degranulation were examined in vitro. Mast cells were treated with suspensions of PM_2.5 in Dulbecco’s modified Eagle’s medium at concentrations from 25 to 200?mg/L in the absence or presence of 10?mmol/L N-acetyl-L-cysteine. Biological effects and mitochondrial function were assessed by determining cell viability, β-hexosaminidase release, interleukin-4 secretion, reactive oxygen species generation, adenosine triphosphate production, potential alteration of mitochondrial membrane, and activities of mitochondrial electron transport chain complexes I and III. Exposure of mast cells to PM_2.5 induced reduction of adenosine triphosphate production, collapse of mitochondrial membrane potential, and inhibition of the activity of complex III. Co-treatment of mast cells exposed to PM_2.5 with N-acetyl-L-cysteine attenuated cytotoxicity and the production of reactive oxygen species, and decreased the release of β-hexosaminidase and interleukin-4. Evidently, PM_2.5-induced oxidative stress plays an essential role in mitochondrial toxicity and mast cell activation.     

Microbial fuel cells: Advances in electrode modifications for improvement of system performance

There is an increasing trend to conduct the researches related to Microbial Fuel Cells (MFCs) in the recent years. Limited power output has been the major obstacle for the practical application and upscaling of MFCs. Attempts have been made on electrode modifications such as anode treatments, cathode modifications with catalysts and bio cathodes developments to produce varying degrees of improved output current depending upon the types of modifications. Power density and Coulombic efficiency have been considered as the important parameters to analyze the system performances. This paper overviews on the advances made in MFCs’ researches focusing on different types of electrodes modifications along with the involved methodology. Furthermore, the system performances of different modified MFCs are compared in terms of the power density and the Coulombic efficiency.     

Toxicity of pesticides toward human immune cells U-937 and HL-60

Abstract

The industrialization of the agricultural sector has significantly increased the use of chemicals such as pesticides. Therefore, exposure to them is unavoidable, which makes it necessary to assess their safety for humans at actual exposure doses. This paper aims to determine toxicity of three types of pesticides toward human immune cells (HL-60 and U-937): glyphosate (GLY), deltamethrin (DEL), and chlorothalonil (CHL). Cell viability, membrane integrity, inflammation induction, and antioxidant activity were evaluated to determine differences in cellular response to the tested plant protection agents. In experimental models, all tested substances caused increased mortality of cells after only 24?h. Cell membrane damage was recorded under DEL and CHL influences. The largest disintegration of the cell membrane was due to the action of 100?μg/mL DEL for U-937 and CHL at 1?μg/mL for HL-60. GLY at a concentration of 3,600?μg/mL caused significant peroxidation of U-937 cells’ lipids. CHL-induced inflammation in both types of cells tested. DEL and GLY also induced antioxidant activity in cells. These results lead to the conclusion that the tested pesticides act cytotoxically to the cells of the human immune system in doses to which both farmers and consumers are exposed.     

长江、嘉陵江(重庆段)源水有机提取物的类雌激素活性评价

为了解长江、嘉陵江(重庆段)源水有机提取物的类雌激素活性，分别于夏季和冬季采用GDX—120大孔树脂对位于城区上、中、下游5个断面长江、嘉陵江(重庆段)源水进行有机物的浓缩提取。用体外MCF—7细胞增殖试验检测有机提取物的类雌激素活性。结果显示，源水有机提取物均具有类雌激素活性。其类雌激素活性中、下游均高于上游，丰水期高于枯水期。