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.     

焚烧飞灰重金属含量及浸出长期变化规律研究

通过对24个飞灰样品的重金属全量消解和高精度X射线荧光光谱法（HDXRF）快速检测结果进行线性回归分析,发现HDXRF法可较准确测定飞灰中Cu、Pb、Zn、Ni和Cd含量（线性回归决定系数R²³0.90）.采用HDXRF法对某焚烧厂飞灰重金属含量6个月快速检测及浸出毒性测定数据统计分析发现：飞灰重金属月均含量较稳定（变异系数CV<0.14）但日均含量波动较大（CV>2.54）;飞灰中Pb、Cu和Zn含量呈正相关（相关系数r>0.78）;金属浸出浓度受浸出液终点pH值影响显著,超标频率依次为Pb（68.9%）、Cd（66.1%）、Ni（24.0%）、Cu（14.2%）和Zn（1.6%）.短期飞灰金属含量和浸出毒性数据受焚烧工况,烟气处理工况和垃圾组分等多种因素共同影响,不具有代表性.为评估垃圾分类源头削减重金属效果,建议采用快速检测方法对焚烧飞灰重金属含量进行长期监测.     

An investigation of methods for enriching trophoblast from maternal blood

Trophoblast deportation is known to occur in normal human pregnancy, but it is not yet clear whether these cells routinely enter the maternal peripheral circulation and are available as a source of fetal DNA for non-invasive prenatal diagnosis of genetic disorders. To resolve this issue requires an efficient method of enriching trophoblast from maternal blood combined with a means to confirm its identity. Five different techniques were tested on ten retroplacental blood samples to determine the most sensitive and operator-efficient method. Lysis of red cells alone gave the best recovery of trophoblast but had to be discounted, together with Ficoll density gradient centrifugation, due to the very low purity and the excessive time required. Fluorescence-activated cell sorting (FACS) of pre-enriched trophoblast resulted in the lowest recovery rate (8 per cent) despite a 3250-fold enrichment and a very high purity. Immunomagnetic beads (Dynabeads) coated with anti-CD 16 antibody proved to be the best method for the subsequent immunocytochemical characterization of deported trophoblast. However, IO beads coated with anti-CD45 antibody may be more useful for isolating trophoblast for prenatal diagnosis due to the high purity, enrichment (32-fold), and recovery rate (78 per cent) obtained with this method.     

Flow sorting of fetal erythroblasts using intracytoplasmic anti-fetal haemoglobin: Preliminary observations on maternal samples

Monoclonal antibody to fetal haemoglobin (a₂γy₂) has been proposed as a fetal-specific reagent. We developed an intracellular staining protocol that combines fluorescein isothiocyanate or phycoerythrin conjugated anti-γ with the DNA binding dye Hoechst 33342 to identify and flow sort fetal erythroblasts from maternal blood. Our preliminary observations on anti-γ-positive cells sorted from four different pregnant women are described here, using fluorescence in situ hybridization (FISH) with chromosome-specific probes to identify fetal cells. Our data demonstrate that far fewer candidate fetal cells are sorted with this protocol than by current cell surface staining methods that employ the monoclonal antibody CD71. This results in increased fetal cell sorting purities. With this protocol, standard FISH techniques require modification due to the rigorous fixation with 4 per cent paraformaldehyde. Our initial data indicate the promise of this approach.     

建筑安全风险因素分级排序的风险矩阵法

为减少风险分级排序时产生的风险结,在引入施工安全事故频率的基础上,采用单元格细化和Borda序值法改进原有的风险矩阵法。改进后的方法使得风险等级数量增加,风险结数量减少。将该方法应用于脚手架工程的风险因素的分级与排序,风险等级分为6级,且实现了12个因素的明确排序。     

北京核与辐射恐怖事件风险分析

采用风险分类-要素分析-风险排序的思路,对北京市核与辐射恐怖事件风险进行分析。根据国际上1998—2006年该类恐怖事件的实例,从事件所涉及的材料、场所角度将核与辐射恐怖事件分为6大类。结合北京市的实际情况进行涉核恐怖事件的要素分析,将北京市各类场所划分为4类敏感区并与5类涉核恐怖事件结合起来,得到北京市核与辐射恐怖事件风险构成图。通过风险分析,笔者认为北京市发生核与辐射恐怖事件风险最大的场所为核研究机构,最容易发生的核与辐射恐怖事件类型为涉及放射源的恐怖事件,结合分析结果对从阻止、探测、响应3个层次上提出了奥运核安保工作的反核恐工作建议。     

防洪物资储备决策方法初探

根据防洪抢险物资储备属于一种信息不全型决策问题的特点,本文提出了一种用于确定每年防灾物资准备量概率排序决策模型,并以孙不河流泥石流灾害预报火灾决策为例进行说明。     

城市生活垃圾综合处理前分选系统设计

本文以江油市城市生活垃圾处理厂为工程实例,介绍了垃圾前分选系统中的垃圾储料车间、行车与抓斗、垃圾上料机构、双层振动筛以及人工分选平台的工艺设计参数及设计依据,并在此基础上提出了该系统的改进建议.     

The habitat saturation hypothesis and sociality in an allodapine bee: cooperative nesting is not “making the best of a bad situation”

Several factors thought to be important for the evolution of cooperative breeding in vertebrates have received little attention in facultatively social insects. One of these, the “habitat saturation hypothesis” of Selander (1964), predicts that colony sizes will be greater in breeding units where dispersal opportunities are limited, suggesting that group living is a secondary option to independent reproduction. The Australian allodapine bee Exoneura bicolor exhibits a number of traits that occur in cooperatively breeding bird species, including long life-span, repeated opportunities for reproduction, and vulnerability to brood predation and parasitism. We experimentally examined the effect of a potentially limiting environmental factor, nesting substrate availability, as an agent influencing sociality in E. bicolor. We manipulated nesting substrate availability in two separate locations during a time when foundress dispersal is common. No significant difference was found between colony sizes in cases where dispersal options were abundant and cases where dispersal options were limited. An increase in opportunities for dispersal did not lead to higher rates of independent nesting, suggesting that cooperative nesting is a preferred strategy regardless of distance-related costs of dispersal. Reproductivity per female and brood survival were examined as factors selecting for group living. Low survival of brood in single-female nests has the potential to select for cooperative nesting in this bee. Received: 29 September 1995/Accepted after revision: 24 June 1996