Objective: To establish a prenatal non-invasive method for combined detection of fetal ABO, RhD, and RhCE blood group genotypes based on fluorescence quantitative PCR (FQ-PCR) technology, and to evaluate its clinical value in the early prenatal diagnosis of hemolytic disease of the fetus and newborn (HDFN). Methods: A total of 200 high-risk singleton pregnant women who underwent prenatal examinations in our hospital from January 2022 to December 2024 were prospectively enrolled. They were divided into four groups: the ABO incompatibility group (n=100), the RhD incompatibility group (n=50), the RhCE incompatibility group (n=50), and the control group (n=200). FQ-PCR technology was used to detect cell-free fetal DNA (cff-DNA) in maternal plasma, targeting the ABO system (261delG, 796C>A), exons 5/7 of the RHD gene, and the key loci of RhCE system (C/c, E/e). After delivery, the blood group of newborn was verified by serological testing of umbilical cord blood., and the hemolysis panel tests (direct antiglobulin test, free antibody test, and antibody release test) were performed to evaluate the detection consistency and identify high-risk factors. Results: The detection coincidence rates for ABO, RhD, and RhCE blood groups were 98.0% (98/100), 100.0% (50/50), and 96.0% (48/50), respectively. The incidence of HDFN in the ABO incompatibility group was 69.0% (69/100), which is significantly higher than that in the RhD incompatibility group (10.0%, 5/50) and the RhCE incompatibility group (2.0%, 1/50). Multivariate analysis identified maternal blood type O (OR=3.021), maternal RhD-negative (OR=5.253), and maternal age ≥35 years (OR=1.950) as independent risk factors for HDFN (all P<0.05). Conclusion: Prenatal non-invasive combined detection of multiple blood group antigen genotypes can significantly improve the efficiency of early diagnosis of HDFN and provide accurate early warning for high-risk pregnant women.

Objective To investigate the changes of T cell subsets including Th1,Th2,Th17 and Treg in the hand,foot and mouth disease(HFMD) patients.Methods 35 EV71 induced HMFD patients and 12 non-infectious children were included in this study.Patients were divided into mild symptoms group and severe symptoms group.The ratios of Th1,Th2,Th17 and Treg were analyzed by flow cytometry.Results Sixteen mild cases and 19 severe cases were included in the 35 patients.No statistical difference of children′s age and sex were found among the groups(P>0.05).The proportion[M(P25,P75)] of Th1 cells of mild and severe groups were 10.65%(8.73%,14.08%) and 15.51% (13.47%,18.31%),respectively,which were significantly higher than normal control group 5.51% (3.61%,8.61%)(P<0.05).The proportion of Th2 cells of mild and severe groups were 4.32%(2.60%,5.32%) and 3.87% (2.34%,5.32%),no significant difference from control group 2.11% (1.02%,5.11%)(P<0.05).The proportion of Th17 cells of mild and severe groups were 8.32%(5.25%,12.33%),7.36% (5.05%,11.9%),significantly higher than that of normal control group 4.23% (2.44%,6.22%)(P<0.05).The proportion of Treg cells of mild and severe groups were 1.46% (0.31%,2.73%),1.59%(0.35%,2.62%),no significant difference from the control group 1.78% (0.31%,2.77%)(P<0.05).Conclusion The Th1/Th2 and Th17/Treg ratios of HFMD patients with EV71 virus infection were changed,and the Th1 and Th17 cells may play an important role in the in the disease and body′s resistance.