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.     

Extraction of DNA from amniotic fluid cells for the early prenatal diagnosis of genetic disease

Ten-ml samples of amniotic fluid were taken from pregnancies being terminated at 8–14 weeks' gestation. DNA was extracted from the amniotic cells by sequential centrifugation and analysed using the polymerase chain reaction (PCR). Fifteen samples were analysed for evidence of maternal contamination using Mfd5 oligo-nucleotide primers for repeat polymorphisms. Ten amniotic fluid samples were tested for the Delta-F₅₀₈ deletion characteristic of cystic fibrosis to demonstrate a diagnostic application for the technique. In each case, DNA extracted from fetal tissue from the same pregnancy was included in the controls. In 14 of the 15 cases tested with the Mfd5 primers, both the amniotic fluid DNA and the fetal DNA showed no evidence of contaminating DNA. In one case, neither the amniotic fluid cells nor the fetal cells yielded results. In nine of the ten cases tested with the Delta-F₅₀₈ primers, the amniotic fluid cell DNA provided accurate information about the genetic status of the fetus; in the tenth, the fetal DNA failed to amplify. The results indicate that adequate DNA can be extracted from amniotic fluid from 8 weeks' gestation onward and these samples are suitable for prenatal diagnosis using PCR.     

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.     

生物传感器快速测定BOD在海洋监测中的应用

选用一株耐高渗透压的酵母菌种作为敏感材料可以实现BOD的快速测定。采用海藻酸钙包埋法和聚乙烯醇包埋法两种细胞固定化方法并进行了比较。就聚乙烯醇固定化微生物进行了渗稼压和重金属离子影响的研究。     

固定化紫色非硫光合细菌降解活性艳红X-3B的研究

采用聚集－交联法固定化紫色非硫光合细菌用于处理活性艳红Ｘ－３Ｂ染液。比较紫色非硫光合细菌和固定化细胞的某些性质,２种细胞的反应最适温度均为３０－４０℃,固定化细胞的反应最适ｐＨ值范围较宽,为７．５－９．４,热稳定性较好。Ｃｕ（２＋）对２种细胞酶活力均有抑制作用;比较聚集－交联固定化细胞和海藻酸钠包埋固定化细胞的脱色能力,前者比后者酶活力较高,半衰期长,成本低,操作方法简单,易于工业化应用。     

Prenatal detection of trisomy 21 in uncultured amniocytes by fluorescence in situ hybridization: A prospective study

A prospective study was undertaken to evaluate the use of fluorescence in situ hybridization (FISH) for the detection of trisomy 21 in interphase nuclei of uncultured amniotic fluid cells. Five hundred cases were analysed in situ and classified as normal or abnormal; the results were subsequently checked against the cytogenetic findings. Four hundred and ninety-three were correctly identified as normal with an 86·6 per cent average frequency of scored nuclei exhibiting two signals; six cases were correctly identified as trisomic for chromosome 21 with 81·7 per cent of scored nuclei exhibiting three signals; and one abnormal case involving an unbalanced chromosome 21·21 translocation was falsely scored as normal due to poor hybridization/detection efficiency. The method has been substantially improved and simplified so that it is suitable for the rapid detection of trisomy 21. As aneuploidy detection in interphase does not identify structural chromosome aberrations, it is not a substitute for fetal chromosome analysis.     

Rapid prenatal diagnosis of aneuploidy from uncultured amniotic fluid cells using five-colour fluorescence in situ hybridization

In this paper we describe the use of five-colour fluorescence in situ hybridization for prenatal diagnosis of aneuploidy using uncultured amniotic fluid cells. The analysis is based on ratio mixing of dual-labelled probes and digital imaging for the detection and visualization of five different probes specific for the five target chromosomes, 13, 18, 21, X, and Y. A retrospective blind analysis of 30 coded uncultured amniotic fluid samples correctly detected fetal sex and five trisomy 21 cases. Multicolour fluorescence in situ hybridization used in this way allows rapid and simultaneous detection of the most frequent aneuploidies.     

大气飘尘对大鼠肺细胞的毒性比较

采用大白鼠气管内注入飘尘后分析肺灌洗液的方法，比较了上海不同地区飘尘对大鼠肺细胞的毒性；再运用毒性指数及效应指数，综合评价了上海市不同地区大气飘尘污染的状况。研究表明：从肺灌洗液的生化成份分析及细胞分类显示，以彭浦新村、隆昌路平凉路、康定路胶州路３个点的飘尘毒性较大。综合评价结果，彭浦新村的效应指数最大，以后依次为康定路胶州路、鲁班路斜土路、青云路、隆昌路平凉路等。     

食管癌高发区粮食防霉实验

磁县是我国食管癌高发区,当地居民饮食中霉菌及其毒素污染严重。为了从病因水平上预防肿瘤发生,进一步探讨了磁县食管癌高发区居民饮食防霉措施。本实验研究了防腐剂富马酸二甲酯（DMF）对磁县农户正在食用的储粮的抑霉防霉及抑菌作用。结果发现DMF可明显抑制粮食中霉菌的生长。小麦洗液培养56小时后DMF处理组霉菌菌落数平均为2.3个,而对照组可达123.7个,抑制率为95.15％。在含有DMF的培养基上霉菌不能生长。此外DMF还有明显抑菌作用,加DMF后48h即可抑制细菌生长,施药处出现明显的抑菌环。因此,应在食管癌高发区对农村储粮推广使用DMF防霉措施,以期通过降低霉菌及其毒素的污染,阻断食管癌的霉菌病因,进一步降低食管癌发病率。