This study examined the methylation difference in AIRE and RASSF1A between maternal and placental DNA, and the implication of this difference in the identification of free fetal DNA in maternal plasma and in prenatal ...This study examined the methylation difference in AIRE and RASSF1A between maternal and placental DNA, and the implication of this difference in the identification of free fetal DNA in maternal plasma and in prenatal diagnosis of trisomy 21. Maternal plasma samples were collected from 388 singleton pregnancies, and placental or chorionic villus tissues from 112 of them. Methylation-specific PCR （MSP） and methylation-sensitive restriction enzyme digestion followed by fluorescent quantitative PCR （MSRE ＋ PCR） were employed to detect the maternal-fetal methylation difference in AIRE and RASSF1A. Diagnosis of trisomy 21 was established according to the ratio of fetal-specific AIRE to RASSF1A in maternal plasma. Both methods confirmed that AIRE and RASSF1A were hypomethylated in maternal blood cells but hypermethylated in placental or chorionic villus tissues. Moreover, the differential methylation for each locus could be seen during the whole pregnant period. The positive rates of fetal AIRE and RASSF1A in maternal plasma were found to be 78.1% and 82.1% by MSP and 94.8% and 96.9% by MSRE ＋ PCR. MSRE ＋ PCR was superior to MSP in the identification of fetal-specific hypermethylated sequences （P〈0.05）. Based on the data from 266 euploidy pregnancies, the 95% reference interval of the fetal AIRE/RASSF1A ratio in maternal plasma was 0.33-1.77, which was taken as the reference value for determining the numbers of fetal chromosome 21 in 102 pregnancies. The accu-racy rate in 98 euploidy pregnancies was 96.9% （95/98）. Three of the four trisomy 21 pregnancies were confirmed with this method. It was concluded that hypermethylated AIRE and RASSF1A may serve as fetal-specific markers for the identification of fetal DNA in maternal plasma and may be used for noninvasive prenatal diagnosis of trisomy 21.展开更多
A novel kind of multi-core magnetic composite particles, the surfaces of which were respectively mo- dified with goat-anti-mouse IgG and antitransferrin receptor（anti-CD71）, was prepared. The fetal nucleated red blo...A novel kind of multi-core magnetic composite particles, the surfaces of which were respectively mo- dified with goat-anti-mouse IgG and antitransferrin receptor（anti-CD71）, was prepared. The fetal nucleated red blood cells（FNRBCs） in the peripheral blood of a gravida were rapidly and effectively enriched and separated by the mo- dified multi-core magnetic composite particles in an external magnetic field. The obtained FNRBCs were used for the identification of the fetal sex by means of fluorescence in situ hybridization（FISH） technique. The results demonstrate that the multi-core magnetic composite particles meet the requirements for the enrichment and speration of FNRBCs with a low concentration and the accuracy of detetion for the diagnosis of fetal sex reached to 95%. Moreover, the obtained FNRBCs were applied to the non-invasive diagnosis of Down syndrome and chromosome 3p21 was de- tected. The above facts indicate that the novel multi-core magnetic composite particles-based method is simple, relia- ble and cost-effective and has opened up vast vistas for the potential application in clinic non-invasive prenatal diag- nosis.展开更多
Non-invasive prenatal gene diagnosis has been developed rapidly in the recent years, and numerous medical researchers are focusing on it. Such techniques could not only achieve prenatal diagnosis accurately, but also ...Non-invasive prenatal gene diagnosis has been developed rapidly in the recent years, and numerous medical researchers are focusing on it. Such techniques could not only achieve prenatal diagnosis accurately, but also prevent tangential illness in fetuses and thus, reduce the incidence of diseases. Moreover, it is non-invasive prenatal gene diagnosis that prevents potential threaten and danger to both mothers and fetuses. Therefore, it is welcomed by clinical gynecologist and obstetrian, researchers of medical genetics, and especially, pregnancies. This review article touches briefly on the advanced development of using cell-free DNA, RNA in maternal plasma and urine for non-invasive prenatal gene diagnosis.展开更多
Conventional prenatal diagnosis relies on invasive chorionic biopsy or amniocentesis, which increases the risk of miscarriage, and is undertaken at 11-20 weeks gestation.1 The discovery of cell-free fetal DNA in mater...Conventional prenatal diagnosis relies on invasive chorionic biopsy or amniocentesis, which increases the risk of miscarriage, and is undertaken at 11-20 weeks gestation.1 The discovery of cell-free fetal DNA in maternal plasma has, however, offered a new strategy for non-invasive prenatal diagnosis.2 Cell-free fetal DNA in maternal plasma has been used for the determination of fetal gender3 and RHD status4 as well as testing certain monogenic diseases such as 13-thalassemia5 and cystic fibrosis.6 However,展开更多
目的采用N-乙酰半乳糖(N-Acetyl-D-galactosamine,GalNAc)包被的玻片分离和富集胎儿有核红细胞(fetalnucleated red blood cells,FNRBCs)。方法用密度梯度离心法收集胎儿脐血中的FNRBCs,然后将其与不同浓度的大豆凝聚素(SBA)混合,所形...目的采用N-乙酰半乳糖(N-Acetyl-D-galactosamine,GalNAc)包被的玻片分离和富集胎儿有核红细胞(fetalnucleated red blood cells,FNRBCs)。方法用密度梯度离心法收集胎儿脐血中的FNRBCs,然后将其与不同浓度的大豆凝聚素(SBA)混合,所形成的“FNRBCs-SBA”复合物用GalNAc包被的玻片来分离和富集,分析SBA工作浓度对FNRBCs分离和富集效率的影响。结果当GalNAc包被玻片的浓度为100μg/ml时,20μg/ml的SBA浓度是分离和富集FNRBCs的最适浓度;此时WBC与有核红细胞的比值较低(27.7%),而FNRBCs与有核红细胞的比值最高(3.7%)。结论N-乙酰半乳糖特异性植物凝素可用于选择性地分离和富集FNRBCs,与传统方法相比较具有无需特殊仪器与设备,简便快速的优点。展开更多
目的:探讨噻托溴铵联合康复锻炼对慢性阻塞性肺疾病(Chronic obstructive pulmonary disease,COPD)稳定期患者血清IL-8、肺功能及生活质量的影响。方法:选取2014年2月至2015年2月在我院就诊的84例COPD稳定期患者作为研究对象,按随机数...目的:探讨噻托溴铵联合康复锻炼对慢性阻塞性肺疾病(Chronic obstructive pulmonary disease,COPD)稳定期患者血清IL-8、肺功能及生活质量的影响。方法:选取2014年2月至2015年2月在我院就诊的84例COPD稳定期患者作为研究对象,按随机数字表法将其分为对照组和观察组,每组各42例。对照组患者予以口服氨茶碱释缓片、噻托溴铵吸入进行治疗,观察组患者在此基础上加用康复锻炼,治疗一个月后,比较两组患者血清IL-8、第1s用力呼气量(Forced expiratory volume in the first second,FEV1)、第1s用力呼气容积占预测值百分比(Forced expiratory volume in the first second/forced vital capacity,FEV1/FVC)、生活质量等指标。结果:(1)两组患者治疗后血清IL-8水平较治疗前明显降低,且观察组降低程度明显大于对照组(P<0.05);(2)两组患者治疗后FEV1、FEV1/FVC等肺功能指标均较治疗前提高,且观察组提高程度明显大于对照组(P<0.05);(3)两组患者治疗后生理机能比较无明显差异(P>0.05),观察组患者生理职能、躯体疼痛、一般健康、精力、社会功能、情感职能以及精神健康评分均高于对照组(P<0.05)。结论:噻托溴铵联合康复锻炼对COPD稳定期患者具有良好的临床疗效,优于单纯噻托溴铵治疗,可有效改善患者血清IL-8、肺功能指标,提高患者生活质量,值得临床推广应用。展开更多
基金supported by grants from Health Department of Hubei Province (No. QJX2008-28)Science and Technology Bureau of Wuhan (No. 200760423158)Population and Family Planning Commission of Wuhan, China (No. WRJK0906)
文摘This study examined the methylation difference in AIRE and RASSF1A between maternal and placental DNA, and the implication of this difference in the identification of free fetal DNA in maternal plasma and in prenatal diagnosis of trisomy 21. Maternal plasma samples were collected from 388 singleton pregnancies, and placental or chorionic villus tissues from 112 of them. Methylation-specific PCR （MSP） and methylation-sensitive restriction enzyme digestion followed by fluorescent quantitative PCR （MSRE ＋ PCR） were employed to detect the maternal-fetal methylation difference in AIRE and RASSF1A. Diagnosis of trisomy 21 was established according to the ratio of fetal-specific AIRE to RASSF1A in maternal plasma. Both methods confirmed that AIRE and RASSF1A were hypomethylated in maternal blood cells but hypermethylated in placental or chorionic villus tissues. Moreover, the differential methylation for each locus could be seen during the whole pregnant period. The positive rates of fetal AIRE and RASSF1A in maternal plasma were found to be 78.1% and 82.1% by MSP and 94.8% and 96.9% by MSRE ＋ PCR. MSRE ＋ PCR was superior to MSP in the identification of fetal-specific hypermethylated sequences （P〈0.05）. Based on the data from 266 euploidy pregnancies, the 95% reference interval of the fetal AIRE/RASSF1A ratio in maternal plasma was 0.33-1.77, which was taken as the reference value for determining the numbers of fetal chromosome 21 in 102 pregnancies. The accu-racy rate in 98 euploidy pregnancies was 96.9% （95/98）. Three of the four trisomy 21 pregnancies were confirmed with this method. It was concluded that hypermethylated AIRE and RASSF1A may serve as fetal-specific markers for the identification of fetal DNA in maternal plasma and may be used for noninvasive prenatal diagnosis of trisomy 21.
文摘A novel kind of multi-core magnetic composite particles, the surfaces of which were respectively mo- dified with goat-anti-mouse IgG and antitransferrin receptor（anti-CD71）, was prepared. The fetal nucleated red blood cells（FNRBCs） in the peripheral blood of a gravida were rapidly and effectively enriched and separated by the mo- dified multi-core magnetic composite particles in an external magnetic field. The obtained FNRBCs were used for the identification of the fetal sex by means of fluorescence in situ hybridization（FISH） technique. The results demonstrate that the multi-core magnetic composite particles meet the requirements for the enrichment and speration of FNRBCs with a low concentration and the accuracy of detetion for the diagnosis of fetal sex reached to 95%. Moreover, the obtained FNRBCs were applied to the non-invasive diagnosis of Down syndrome and chromosome 3p21 was de- tected. The above facts indicate that the novel multi-core magnetic composite particles-based method is simple, relia- ble and cost-effective and has opened up vast vistas for the potential application in clinic non-invasive prenatal diag- nosis.
文摘Non-invasive prenatal gene diagnosis has been developed rapidly in the recent years, and numerous medical researchers are focusing on it. Such techniques could not only achieve prenatal diagnosis accurately, but also prevent tangential illness in fetuses and thus, reduce the incidence of diseases. Moreover, it is non-invasive prenatal gene diagnosis that prevents potential threaten and danger to both mothers and fetuses. Therefore, it is welcomed by clinical gynecologist and obstetrian, researchers of medical genetics, and especially, pregnancies. This review article touches briefly on the advanced development of using cell-free DNA, RNA in maternal plasma and urine for non-invasive prenatal gene diagnosis.
文摘Conventional prenatal diagnosis relies on invasive chorionic biopsy or amniocentesis, which increases the risk of miscarriage, and is undertaken at 11-20 weeks gestation.1 The discovery of cell-free fetal DNA in maternal plasma has, however, offered a new strategy for non-invasive prenatal diagnosis.2 Cell-free fetal DNA in maternal plasma has been used for the determination of fetal gender3 and RHD status4 as well as testing certain monogenic diseases such as 13-thalassemia5 and cystic fibrosis.6 However,
文摘目的采用N-乙酰半乳糖(N-Acetyl-D-galactosamine,GalNAc)包被的玻片分离和富集胎儿有核红细胞(fetalnucleated red blood cells,FNRBCs)。方法用密度梯度离心法收集胎儿脐血中的FNRBCs,然后将其与不同浓度的大豆凝聚素(SBA)混合,所形成的“FNRBCs-SBA”复合物用GalNAc包被的玻片来分离和富集,分析SBA工作浓度对FNRBCs分离和富集效率的影响。结果当GalNAc包被玻片的浓度为100μg/ml时,20μg/ml的SBA浓度是分离和富集FNRBCs的最适浓度;此时WBC与有核红细胞的比值较低(27.7%),而FNRBCs与有核红细胞的比值最高(3.7%)。结论N-乙酰半乳糖特异性植物凝素可用于选择性地分离和富集FNRBCs,与传统方法相比较具有无需特殊仪器与设备,简便快速的优点。
文摘目的:探讨噻托溴铵联合康复锻炼对慢性阻塞性肺疾病(Chronic obstructive pulmonary disease,COPD)稳定期患者血清IL-8、肺功能及生活质量的影响。方法:选取2014年2月至2015年2月在我院就诊的84例COPD稳定期患者作为研究对象,按随机数字表法将其分为对照组和观察组,每组各42例。对照组患者予以口服氨茶碱释缓片、噻托溴铵吸入进行治疗,观察组患者在此基础上加用康复锻炼,治疗一个月后,比较两组患者血清IL-8、第1s用力呼气量(Forced expiratory volume in the first second,FEV1)、第1s用力呼气容积占预测值百分比(Forced expiratory volume in the first second/forced vital capacity,FEV1/FVC)、生活质量等指标。结果:(1)两组患者治疗后血清IL-8水平较治疗前明显降低,且观察组降低程度明显大于对照组(P<0.05);(2)两组患者治疗后FEV1、FEV1/FVC等肺功能指标均较治疗前提高,且观察组提高程度明显大于对照组(P<0.05);(3)两组患者治疗后生理机能比较无明显差异(P>0.05),观察组患者生理职能、躯体疼痛、一般健康、精力、社会功能、情感职能以及精神健康评分均高于对照组(P<0.05)。结论:噻托溴铵联合康复锻炼对COPD稳定期患者具有良好的临床疗效,优于单纯噻托溴铵治疗,可有效改善患者血清IL-8、肺功能指标,提高患者生活质量,值得临床推广应用。
