Detection of both the normal and mutant alleles in single cells of individuals heterozygous for the sickle cell mutation—prelude to preimplantation diagnosis

As a preliminary step to preimplantation diagnosis of sickle cell disease in unfertilized eggs or 8-cell embryos of heterozygous parents, we established quality control for detection of the mutant and normal alleles of the beta-haemoglobin gene using single buccal cells. Efficient polymerase chain reaction (PCR) amplification of a 680 base pair sequence of the beta-globin gene spanning the site of the sickle cell mutation was obtained for 79 per cent of single heterozygous cells. In 71 per cent of cases, both alleles were detected. With this current efficiency, we predict that a clinical preimplantation diagnosis at the 8-cell embryo stage could be carried out safely and reliably for a couple at risk of transmitting sickle cell disease to their children.     

Erythroid-specific antibodies enhance detection of fetal nucleated erythrocytes in maternal blood

Fetal nucleated erythrocytes (NRBC) in maternal blood are a non-invasive source of fetal DNA for prenatal genetic screening. We compared the effectiveness of three monoclonal antibodies for the separation of fetal cells from maternal blood by flow sorting. Mononuclear blood cells from 49 healthy pregnant women were incubated with antibody to CD 71, CD 36, and/or glycophorin A (GPA), employed singly or in combination with each other. These monoclonal antibodies recognize surface antigens on haematopoietic precursor cells. Successful isolation of fetal cells was defined as detection of Y chromosomal sequences in maternal blood from women carrying male fetuses, with absence of Y sequences when female fetuses were carried. Thus, gender prediction accuracy was used as a measure of fetal cell separation. Using anti-CD 71 to isolate fetal cells, gender prediction was 57 per cent correct; with anti-CD 36, it was 88 per cent correct. Anti-GPA, an erythrocyte-specific antigen, used alone or in combination with anti-CD 71 or 36, improved gender prediction to 100 per cent. We conclude that antibody to GPA improves the retrieval of fetal NRBC from maternal blood, permitting genetic analysis by the polymerase chain reaction.     

乐果对金鱼的急性毒性影响

用改进的寇氏法（karber）测定了乐果对金鱼（Carassius auratus）的急性毒性。急性试验经150mg／L、188．4mg／L、234．4mg／L、291．7mg／L、360mg／L乐果暴露4d，在15％～20％下，24、48、72、96h的L11m值分别为297．17、289．13、259．12、238．67mg／L，其95％置信限分别为284．25～310．67、275．17～303．81、243．33～275．93、223．98～254．33mg／L。     

饮用水源中三卤甲烷含量及与遗传毒性的相关性研究

通过对某市饮用水源中不同点位的三卤甲烷浓度测试,同时采用SOS/ Umu 试验对水体进行生物遗传毒性分析,结果表明:采用液氯消毒工艺的地表水厂在消毒后水样和用户端水样中检出的三卤甲烷含量较高,而采用二氧化氯消毒的地下水厂的出水中挥发性有机物仅略有上升;进行三卤甲烷浓度和生物遗传毒性之间的相关性分析得知,三卤甲烷和间接遗传毒性的相关性较好.     

Chorionic villus culture for prenatal diagnosis of chromosome defects: Reduction of the long-term cultivation time

We report in detail two series of chorionic villus cultivation for prenatal chromosomal diagnosis. Chorionic villi were sampled from both first- and second-trimester pregnancies. One hundred cultures were treated with trypsin–EDTA for 2 h and collagenase overnight, (method A) and 100 were treated with trypsin–EDTA for 1 h and collagenase for 2 h (method B). Using short-term enzymatic digestion, the cultivation time was reduced from 14 days to 6 days. Sufficient amounts of metaphases of good quality were present in 93 per cent of primary cultures harvested in situ, whereas enough metaphases of sufficiently good quality were in most cases present only after subcultivation of the cultures using method A. The decrease in cultivation time obtained is probably due to a higher yield of viable cells in monocellular suspension, an increased attachment efficiency, and a more rapid attachment of single cells (within 24 h).     

Preconcentration and determination of trace copper(II) in natural water using silica gel modified with 1-(2-pyridylazo)-2-naphthol

Trace copper (II) in water can be preconcentrated using silica gel modified with 1-(2-pyridylazo)-2-naphthol (PAN) and be determined spectrophotometri-cally. The conditions for the preconcentration were studied. The method was applied to both artificial and natural water samples and the results suggest that copper (II) of ppb level in water can be measured accurately after the preconcentration.     

In vitro characteristics of human fetal cells obtained from chorionic villus sampling and amniocentesis

In vitro characteristics of human fetal cells have been investigated after chorionic villus sampling at the first trimester and amniocentesis at the second trimester of pregnancy. Light microscopy revealed heterogeneous morphology of cell types in both the chorionic villus culture and the amniotic fluid cultures. Based on the experiments performed, chorionic villus cells are more sensitive to pronase, trypsin, and versene during subculture and have a higher DNA content per single cell and release more [¹²⁵I]-Beta-human chorionic gonadotropin into culture medium than those found in amniotic fluid cells. The practical applications of this study are discussed.     

钻井泥浆和消油剂对合浦珠母贝的急性毒性效应

采用96h急性毒性试验配合滤食率和微核检测等方法，比较了石油勘探开发用3种钻井泥浆和3种消油剂对合浦珠母贝成贝急性毒性效应。实验结果表明：3种钻井泥浆对合浦珠母贝的半致死浓度均大于10^5mg/L；3种消油剂的半致死浓度分别为1047、5011和2630mg/L；滤食率与钻井泥浆和消油剂的浓度呈负相关；微核细胞率则与之呈正相关。     

废渣──Al(OH)_3凝胶再利用研究

研究了对处理铝材加工含Al3＋的酸性废液过程中产生的二次污染物Al（OH）3凝胶废渣的应用。含水分约90％、80％左右的Al（OH）3凝胶200g分别与14～16mL、22～24mL95％的工业浓硫酸酸反应，可制得浓度为20％～23％的Al（SO4）3溶液，经稀释可用作造纸原料。     

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.