The Study Of The Nuclear Red Blood Cells' Rich By Change The Micro Environment Around The Cells

Objectives:1.To investigate both quantity and morphology properties of Fetal Nuclear Red BloodCells in umbilical cord blood, which could provide the effectiveness of scientific proof ofthe NRBC rich.2.The aim of this study is to investigate the possibility of fetal free-DNA in prenataldiagnosing sex-linked inherited diseases.Methods:1.To study the cells basic characteristics by liquid then half solid Culture using umbilicalcord blood, we compared the numbers of NRBC recovered by the the Ficoll methodafter preliminary concentration with PVDF or by the the Ficoll method after doing someeffect on membrane Ca²⁺ ionic channels using dansheng then with PVDF.2.Urine of pregnant women at weeks of gestation were enriched by 2000rpm/min20min and then examined by nest-PCR.DNA was extracted from enriched maternal urine.The products of the nested-PCR amplification were run by agarose electrophoresis andvisualized by exposure to ultraviolet light, specific positive trips of 418bp and 133bpcould be seen. By using this method, we could determine the fetus sex and make prenataldiagnosis .Its results were consistent with chromosome analysis and clinical examination.Results:1.DAB and K-B staining can distinguish NRBC and other single nucleolus cells, and K-B staining can make more distinguishes between NRBC and FNRBC; there were no balance about the proportion of NRBC among 81 samples, the range of proportion was 1%～54%, the mean was about 8%; the gain of liquid culture was 20%, the results of cell culture were remain to discussion.2. A new highly sensitive nest PCR was developped to detect an SRY gene sequence in maternal urine. Analysis was performed on 42 pregnant women (mean gestational age:weeks). Among them, 32 pregnant women were observed positive results, Among the 10pregnant women were observed negative results. SRY PCR analysis of maternal urinewas in complete concordance with maternal plasma and blood cells.Conclusions:1. PVDF micro membrane could gain the NRBCs and the recovery rate of PVDF was2.04%, so that using dansheng then with PVDF thet recovery rate of PVDF was 3.80%,so dansheng could promote the rate of NRBC'rich, about 1.86multiple.2.Fetal free-DNA were indeed present in maternal urine and appear in the maternal urineearly in the first trimester about 8 weeks, We could simply enrich them by centrifugationas well as make use of theses DNA with nested-PCR technique to identify fetus sex. Thefetal free-DNA in the maternal urine might be very valuable for early non-invasiveprenatal screening for fetus sex and inherited disorders.