Fetal RHD screening from circulating cell-free DNA in RhD-negative pregnant women / 中华围产医学杂志

Objective:To investigate the RHD genotypes of RhD-negative pregnant women and explore the optimum strategy for fetal RHD screening among this population in the region. Methods:This prospective study recruited 33 cases of RhD-negative singleton pregnancies at ≥12 weeks of gestation in Nanjing Drum Tower Hospital from March to November 2021. On the basis of RHD genotyping, quantitative real-time polymerase chain reaction (PCR) was used to amplify the exons 5 and 10 of RHD gene in the circulating cell-free DNA of RhD-negative pregnant women harboring whole RHD gene deletion and RHD-CE(2-9)- D. High-throughput sequencing was performed to detect chr1:25648453 locus from circulating cell-free DNA in plasma of RhD-negative pregnant women harboring RHD 1227A mutation to screen the fetal RhD blood group. Neonatal umbilical cord blood samples were collected for verifying fetal RHD genotyping. Descriptive statistical analysis was used. Results:Whole RHD gene deletion homozygous genotype ( n=20, 60.6%), RHD-CE(2-9) -D/whole RHD gene deletion heterozygous genotype ( n=5, 21.2%), RHD 1227A/whole RHD gene deletion heterozygous genotype ( n=7, 15.2%) and RHD 711delC/whole RHD gene deletion heterozygous genotype ( n=1) were identified in the 33 RhD-negative pregnant women. In the 25 cases with whole RHD gene deletion homozygous genotype or RHD-CE(2-9)- D/whole RHD gene deletion heterozygous genotype, 22 fetuses were RhD-positive and three were RhD-negative based on prenatal screening, which were confirmed by the neonatal serological test results after birth. In the seven cases carrying RHD 1227A/whole RHD gene deletion heterozygous genotype, all fetuses were RhD-positive, which were consistent with the results of serological detection after delivery. The case harboring RHD 711delC/whole RHD gene deletion heterozygous genotype did not receive fetal RHD screening. Conclusions:This study suggests that whole RHD gene deletion homozygous genotype is the most common allele in RhD-negative population in this area, followed by RHD 1227A/whole RHD gene deletion heterozygous genotype and RHD- CE(2-9)- D/whole RHD gene deletion heterozygous genotype. For women with whole RHD gene deletion homozygous genotype, RHD- CE(2-9)- D, or RHD 1227A mutation, fetal RHD screening with quantitative real-time PCR and high-throughput sequencing are important for the management of RhD-negative pregnant women.

Analysis of genetic etiology and related factors in 1 065 women with spontaneous abortions / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic etiology and related factors in 1 065 women with spontaneous abortions.@*METHODS@#All patients have presented at the Center of Prenatal Diagnosis of Nanjing Drum Tower Hospital from January 2018 to December 2021. Chorionic villi and fetal skin samples were collected, and the genomic DNA was assayed by chromosomal microarray analysis (CMA). For 10 couples with recurrent spontaneous abortions but normal CMA results for abortive tissues, non-in vitro fertilization-embryo transfer (IVF-ET) pregnancies and no previous history of live births and no structural abnormalities of the uterus, peripheral venous blood samples were collected. Genomic DNA was subjected to trio-whole exome sequencing (trio-WES). Candidate variants were verified by Sanger sequencing and bioinformatics analysis. Multifactorial unconditional logistic regression analysis was carried out to analyze the factors that may affect chromosomal abnormality in spontaneous abortions, such as the age of the couple, number of previous spontaneous abortions, IVF-ET pregnancy and history of live birth. The incidence of chromosomal aneuploidies in spontaneous abortions during the first trimester was compared in young or advanced-aged patients by chi-square test for liner trend.@*RESULTS@#Among the 1 065 spontaneous abortion patients, 570 cases (53.5%) of chromosomal abnormalities were detected in spontaneous abortion tissues, which included 489 cases (45.9%) of chromosomal aneuploidies and 36 cases (3.4%) of pathogenic/likely pathogenic copy number variations (CNVs). Trio-WES results have revealed one homozygote variant and one compound heterozygote variants in two pedigrees, both of which were inherited from the parents. One likely pathogenic variant was detected in the patient from two pedigrees. Multifactorial unconditional Logistic regression analysis suggested that age of patient was an independent risk factor of chromosome abnormalities (OR = 1.122, 95%CI: 1.069-1.177, P < 0.001), the number of previous abortions and IVF-ET pregnancy were independent protective factors for chromosomal abnormalities (OR = 0.791, 0.648; 95%CI: 0.682-0.916, 0.500-0.840; P = 0.002, 0.001), whilst the age of husband and history of live birth were not (P > 0.05). The incidence of aneuploidies in the abortive tissues has decreased with the number of previous spontaneous abortions in young patients (χ² = 18.051, P < 0.001), but was not significantly correlated with the number of previous spontaneous abortions in advanced-aged patients with spontaneous abortions (P > 0.05).@*CONCLUSION@#Chromosomal aneuploidy is the main genetic factor for spontaneous abortion, though CNVs and genetic variants may also underlie its genetic etiology. The age of patients, number of previous abortions and IVF-ET pregnancy are closely associated with chromosome abnormalities in abortive tissues.

Research progress on fetal RHD genotype screening in RhD-negative pregnant women in Asia / 中华围产医学杂志

RhD-negative pregnant women with an RhD-positive fetus are at risk of hemolytic disease of the fetus and newborn (HDFN), which may lead to fetal/neonatal death. While these would not affect those RhD-negative fetuses. With the advancement of technology in genetics, the administration of anti-D immunoglobulin to women with an RhD-positive fetus could reduce the risk of HDFN. Therefore, non-invasive prenatal testing on fetal RHD genotype plays an important role in the management of RhD-negative pregnant women. Selective usage of anti-D immunoglobulin is important in perinatal management in these women. The non-invasive prenatal screening for fetal RHD gene which has been carried out in Caucasian is not applicable to Asians because of the difference in RHD genotype. In addition to complete or partial RHD gene deletion, point mutations are also common RHD genotypes among Asians. This enlightens us to establish a non-invasive prenatal screening method for Asians. This article reviews the progress of fetal RHD screening in Asian RhD-negative pregnant women.

The 471st case: duodenal ulcer, mucor infection, and elevated IgG4 / 中华内科杂志

Mucor infection is rarely reported in non-immunocompromised population, especially in isolated gastrointestinal tracts. IgG4-related diseases (IgG4-RD) have been recognized in recent years, but secondary causes of IgG4 elevation should be differentiated. We reported a young man with duodenal mass and ulcer and high serum IgG4 level. Histological biopsy of the mass revealed positive mucor mycelium and infiltration of IgG4 positive plasma cells. Serum IgG4 decreased to normal range after surgical resection and systemic antifungal treatment. This case suggests that isolated mucor mycosis infection can develop in the digestive tract and mimics as IgG4-related disease.

10-year Trend of Early Beta Receptor Blocker Application for Acute Myocardial Infarction Patients in Western Rural China / 中国循环杂志

Objective: To assess the trend of early beta receptor blocker (β-blocker) application (with 24h of admission) for acute myocardial infarction (AMI) patients in western rural China from 2001 to 2011. Methods: A 2-stage random sampling design was performed. The 1st stage: a simple random sampling was used to identify participating hospitals and the 2nd stage: a systematic random sampling approach was conducted in 3 specific years of 2001, 2006 and 2011 to take case study for central medical information abstraction. The changing trends and impact factors of early β-blocker application for AMI patients in western rural area were assessed by multivariate model analysis. Results: 35 hospitals were sampled and 33 of them were finally participated. With necessary exclusion, a total of 486 AMI patients without β-blocker contraindication were enrolled for 2 groups: Suitable group, the patients were suitable for early β-blocker application, n=247 and High risk group, the patients with the high risk for shock occurrence, n=239. The application rates for β-blocker within 24h of admission at 2001, 2006 and 2011 in Suitable group were 19.06%, 54.30% and 56.20%, Ptrend=0.0020; in High risk group were 31.53%, 59.49% and 69.62%, Ptrend=0.0001. In Suitable group, the patients with history of hypertension (OR=1.87, 95% CI 1.06-3.29), smoking (OR=1.97, 95% CI 1.11-3.48) or admitted in 2006 (OR=2.93, 95% CI 1.22-7.03) and 2011(OR=4.67, 95% CI 2.06-10.59) had the higher chance to use β-blocker within 24h of admission. Conclusion: Application of β-blocker within 24h of admission in AMI patients presented the increasing trend in western rural China from 2001 to 2011, while there was still difference from the guideline recommendation. Improved normative application of β-blocker is helpful to enhance the quality of care and prognosis in AMI patients.