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11.
为了探究淡水贝类背角无齿蚌鳃细胞的原代培养途径,并阐释其在评价水体Cd2+毒性效应上的潜力,本研究比较了不同酶分解方法(链霉蛋白酶、胰蛋白酶)的鳃细胞存活率差异,并分析了L-15培养基中不同血清浓度(10% FBS、20% FBS)对鳃细胞存活率的影响,细胞置于20℃生化培养箱中培养.进而根据Cd2+对鳃细胞的LC25值设定0.0625、0.125、0.25、0.5和1.0 mg·L-1 5个Cd2+理论浓度梯度,对原代培养的鳃细胞进行24 h暴露,分析了细胞活力、总超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和酸性磷酸酶(AcP)的变化特征.结果表明:0.025%链霉蛋白酶在4℃分解16 h的鳃细胞存活率为98.2%±0.2%,显著高于0.25%胰蛋白酶在26℃分解30 min的89.4%±3.5%鳃细胞存活率(p<0.05);L15培养基加入10% FBS的细胞存活率总体显著高于添加20% FBS的细胞存活率(p<0.05).在上述较佳的分解和血清浓度组合条件下,细胞培养120 h后,其存活率仍高达90.1%±4.7%.鳃细胞活力随着Cd2+浓度的增加而降低,两者之间呈显著的线性负相关(p<0.05);随着Cd2+浓度的增加,SOD和AcP含量总体增加,而CAT含量呈现出"诱导-抑制"的变化趋势.本研究初步建立了较为适宜的背角无齿蚌鳃细胞的原代培养方法,并揭示了其原代培养鳃细胞的细胞活力及SOD、CAT、AcP水平,具有作为评价水环境Cd2+毒性/污染的生物指标的潜力. 相似文献
12.
Farideh Z. Bischoff PhD Dorothy E. Lewis Joe Leigh Simpson Dianne D. Nguyen Jeffrey Scott Wendy Schober Sarah Murrell Sherman Elias 《黑龙江环境通报》1995,15(12):1182-1184
Recovering and analysing fetal erythrocytes from maternal blood is being pursued for non-invasive prenatal genetic diagnosis. We report the observation of 46, XY/47, XXY mosaicism in fetal cells from a woman whose first-trimester chorionic villus sampling (CVS) initially showed only 46, XY. Only after exhaustive (500 cells) analysis were four XXY cells found in cultured villi. 相似文献
13.
Mr. M. T. Rebello G. Hackett J. Smith F. E. Loeffler S. Robson N. Maclachlan R. W. Beard C. H. Rodeck R. Williamson D. V. Coleman C. Williams 《黑龙江环境通报》1991,11(1):41-46
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. 相似文献
14.
Yun-Ling Zheng Maryann Demaria Dongkai Zhen Theresa J. Vadnais Diana W. Bianchi MD 《黑龙江环境通报》1995,15(10):897-905
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. 相似文献
15.
We report a gas chromatographic-mass spectrometric method which allows the very long chain fatty acids content of trophoblastic tissue to be directly measured in samples collected by biopsy between 8 and 11 weeks of gestation. This method has been successfully applied to the detection of fetal Zellweger syndrome in two pregnant women who had previously delivered affected infants. In one of them, increased concentrations of C26:0 (0.254 versus 0.108±0.035 μ/mg proteins) and C24:0 (1.32 versus 0.815±0.325 μ/mg proteins) in trophoblast indicated that the fetus had Zellweger syndrome, a diagnosis confirmed by pathological findings after abortion. In the second case, the pregnancy was allowed to proceed, on the basis of normal concentrations of very long chain fatty acids in trophoblastic tissue, and its outcome was actually a healthy newborn. 相似文献
16.
17.
Routine ultrasound examination at 30 weeks gestation revealed an intrapulmonary cystic mass in an otherwise normal fetus. Following delivery at term, the diagnosis of congenital cystic adenomatoid malformation of the right lung was confirmed, and an elective right middle lobectomy successfully performed at nine days of age. 相似文献
18.
19.
Ossie Geifman-Holtzman Eliezer J. Holtzman Theresa J. Vadnais Vincent E. Phillips Eleanor L. Capeless Diana W. Bianchi MD 《黑龙江环境通报》1995,15(3):261-268
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. 相似文献
20.
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. 相似文献