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.     

磷回收对厌氧/好氧交替式生物滤池蓄磷/除磷的影响

为能提高生物除磷系统的除磷/回收磷效率,研究采用厌氧/好氧交替式生物滤池(AABF)处理低碳磷比废水,观察利用周期性扩增进水碳源进行磷回收时,生物滤池除磷效率及生物膜内微生物特性的变化.通过分析生物滤池除磷效率变化,生物膜多聚物染色、扫描电镜(SEM)观察以及荧光原位杂交(FISH)技术分析生物滤池系统实施周期性磷回收对生物膜内微生物菌群形态与组成的影响.结果发现,生物滤池在先后经历3次生物蓄磷-磷回收(PB-PR)的运行操作过程中,生物滤池除磷效率分别由磷回收前的60.3%、82.9%、86.6%提高到磷回收后的87.2%、91.2%、93.5%;生物滤池生物膜内优势菌群形态逐渐由大球菌演变为小球菌、杆菌与丝状菌;经过连续3个周期的PB-PR操作,生物滤池底部生物膜中聚磷菌所占比例由43%提高至70%;聚磷菌群在生物膜内混合菌群中的比例随着上流式生物滤池高度的增加而不断提高.结果表明,采用周期性的碳源扩增与回收磷的操作,可以提高生物滤池除磷效率;引起生物滤池生物膜内微生物菌群形态与组成发生变化,促进聚磷菌成为优势菌群.     

地铁运行引起的地面振动实测及传播规律分析

对地铁线路某区间段的地面振动进行实测,采集不同测线和不同测点处的水平方向、竖直方向加速度,并转化为傅里叶幅值谱对其进行衰减规律分析。利用公式将加速度记录换算为振级和分频振级,参照相关的国家标准分析不同距离处的振级和频率成分。对比同一加速度记录中的两对开方向列车引起的地面振动,进一步分析振动衰减的规律。所得到的结果可供地面建筑的隔振减振及城市轨道的规划和设计参考。     

Duplex PCR for preimplantation genetic diagnosis (PGD) of spinal muscular atrophy

The main difficulty in developing a molecular diagnosis of spinal muscular atrophy (SMA) resides in the specific genomic structure of the locus. Indeed, two highly homologous survival motor neurone genes, SMN1 and SMN2, are present at the locus. The detection of the homozygous deletion of exons 7 and 8 of the SMN1 gene, which is present in 90 to 98% of the patients, is based on methods highlighting 1 of the 8 nucleotidic mismatches existing between these 2 genes. In order to offer preimplantation genetic diagnosis (PGD) for SMA, we developed a new allele-specific amplification method. The main disadvantage of our previously described strategy resided in the possibility of diagnosing, in case of amplification failure, an unaffected embryo as affected. We present here a new PGD-SMA method. We established the conditions for three different duplex PCRs, allowing the specific detection of the SMN1 gene and one polymorphic marker, either D5S629, D5S1977, or D5S641. Of the 60 to 90 single cells tested, the PCR efficiency varied from 98 to 100% with a complete genotype obtained in a range between 81 and 87% with a global allele drop-out rate of 9%. Such a test was used to perform 1 PGD cycle for which 7 embryos could be analysed. All the embryos were fully diagnosed, six as unaffected and one as affected. Four embryos were transferred, but no pregnancy ensued. Copyright © 2003 John Wiley & Sons, Ltd.