EDTA和腐殖酸对铜在淡水藻(Scenedesmus subspicatus 86.81 SAG)细胞内外分布的影响

讨论了铜在淡水藻（Scenedesmus subspicatus86.81SAG)细胞壁和细胞内分布规律，以及乙二胺四乙酸（EDTA）和水体腐殖酸（FA）存在下对铜分布的影响，研究结果表明，EDTA和FA的存在显著降低了细胞表面铜的吸附量，但是不影响细胞内铜的浓度，对测定数据进行分析。可以认为溶液相铜与EDTA或FA形成的铜络合物是生物无效形态，不参与铜在藻细胞壁上特定位点或生物配体的竞争结合反应。     

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.     

UV-B辐射对小麦叶抗氧化系统的影响

研究了温室种植的小麦在０（ＣＫ）、８．８２ｋＪ／ｍ＾２（Ｔ１）和１２．６ｋＪ／ｍ＾２（Ｔ２）三种剂量的紫外线Ｂ（ＵＶ－Ｂ）辐射下抗氧化系统的变化及其原因。ＵＶ－Ｂ辐射诱导了Ｈ２Ｏ２的合成，还原型抗坏血酸和还原型谷胱甘肽含量的增加、过氧化氢酶、愈创木酚过氧化物酶、抗坏血酸过氧化物酶和谷胱甘肽还原酶活性升高，而类胡萝卜素稍有所降低、超氧歧化酶（ＳＯＤ）活几乎不受ＵＶ－Ｂ辐射影响，分析表明，ＵＶ－Ｂ辐射     

包埋法固定化细胞技术及其在水处理中的应用研究

固定化细胞技术(主要为包埋技术),由于方法简单、条件温和、较高的稳定性及细胞容量,在废水处理中应用广泛.本文主要介绍了近年来固定化细胞技术在污水深度处理中的应用研究现状和发展前景,并对固定化细胞技术在污水深度处理过程中的影响因素进行简要论述.     

硒酸钠和亚硒酸钠对长春花细胞悬浮生长的影响

研究了硒化合物对长春花(C.roseus)细胞悬浮生长的影响.在pH5.5培养介质中,当Se(Ⅵ)>0.5ppm或Se(Ⅳ)>0.6ppm时,将抑制C.roseus细胞悬浮生长,其致死浓度分别是100ppm Se(Ⅵ)和2.0ppm Se(Ⅳ).C.roseus对硒化合物有效强的富集作用.硒化合物的毒性和培养介质的pH值有关.在本试验条件下,硒酸钠和亚硒酸钠对Hg~(2+)无拮抗作用.     

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.     

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).     

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.     

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.