Nonimmune hydrops fetalis (NIHF) has a highly heterogeneous etiology, with genetic factors contributing substantially to its pathogenesis. This study aimed to investigate the chromosomal and monogenic etiologies of NIHF. This single-center retrospective cohort study included 121 pregnancies complicated by NIHF evaluated at a tertiary referral center between August 2020 and August 2023. All cases underwent copy number variation sequencing (CNV-seq), and 95 additionally underwent whole-exome sequencing (WES), both assays were based on next-generation sequencing (NGS) technology. Cases were stratified according to gestational age at diagnosis, maternal age, conception method, and whether NIHF was isolated or non-isolated. Genetic findings and pregnancy outcomes were analyzed and compared across subgroups, with longitudinal follow-up of pregnancy outcomes. Among the 121 NIHF cases, pathogenic or likely pathogenic genetic findings were identified in 62 cases, yielding an overall positivity rate of 51.2

Left–right (LR) asymmetry disorders present a complex etiology, with genetic factors emerging as a primary contributor. This study aims to explore the genetic underpinnings of chromosomal variants and individual genes in fetuses afflicted with prenatal LR asymmetry disorder. Through a retrospective analysis conducted between 2020 and 2023 at Tongji Hospital, Huazhong University of Science and Technology, genetic outcomes of LR asymmetric disorder were scrutinized utilizing copy number variation sequencing (CNV-seq) and whole exome sequencing (WES) methodologies. With a combination of CNV-seq and WES, 5 fetuses in 17 patients with LR asymmetry had chromosomal or genetic variants. CNV-seq revealed a 16p11.2 microdeletion syndrome in a situs inversus fetus presenting pathogenic and a 2q36.3 microduplication syndrome in a fetus with Heterotaxy presenting a variant of uncertain significance (VUS). WES identified NM_198075.4:c.755del in the LRRC56 gene and NM_001454.4:c.865_868dup in the FOXJ1 gene in two situs inversus cases, along with two variants in DNAH5 in two other fetuses. Further bioinformatics scrutiny was conducted to assess the protein structure and function prediction of these variants, ultimately indicating their potential pathogenicity. The study highlights that fetuses with LR asymmetric disorders may have copy number variants, underscoring the significance of mutations in LRRC56 and FOXJ1 in the development of LR asymmetry disorders.

This study aims to investigate the high-risk factors and ultrasonic examination of umbilical cord torsion (UCT)-related prenatal stillbirth, with a specific focus on diagnostic challenges and clinical implications. We retrospectively analyzed the electronic data of 108 pregnant women(after 1:2 age propensity score matching from an initial 424 cases) with UCT who delivered from January 2013 to February 2024 in Tongji Hospital, Tongji Medical College, University of Science and Technology. To mitigate confounding and address group imbalance, a 1:2 matching principle of the same age was performed (36 stillbirths vs. 72 live fetuses). Univariate and multivariate logistic regression analyses were used to identify independent predictors. Model performance was assessed by the area under the receiver operating characteristic curve (AUC), Hosmer-Lemeshow test, and Adjusted R². The diagnostic utility of prenatal ultrasound was comprehensively evaluated. Analysis of the matched cohort confirmed balance in baseline characteristics and revealed seven significant associations. The multivariate model demonstrated excellent performance (AUC = 0.943, 95

ObjectiveSpinal muscular atrophy (SMA) is an autosomal recessive disorder mainly affecting the neuromuscular system, which seriously threatens the life and health of patients. But few studies have reported the acceptance rate of SMA gene screening and SMA carrier rate in China. The present study aimed to clarify the two issues in China through a retrospective analysis of 18,818 reproductive age women in Wuhan area of China.MethodsThe copy number (CN) of exons 7 and 8 in survival motor neuron 1 (SMN1) gene was detected by real-time quantitative PCR, and the results were verified by multiplex ligation-dependent probe amplification.ResultsCarrier screening was offered to 44,953 women of childbearing age in our medical center from March, 2018, to February, 2022, of whom 18,818 were enrolled in the program. A total of 336 women were identified as carriers (1.73%; 326/18,808; without fertility history of the children with SMA). Among 18,818 reproductive age women, 286 spouses (85.12%; 286/336) were successfully recalled for screening. The results showed 17 couples at high risk of having children with SMA, of whom prenatal diagnosis was implemented in 11, and 6 fetuses were identified with SMA. All the 5 pregnant women bearing the 6 SMA fetuses chose to terminate the pregnancy by artificial abortion.ConclusionReproductive age women and their spouses in Wuhan area showed a positive attitude toward general screening for SMA carriers. Given the high early mortality of children with SMA, screening for SMA carriers in women of reproductive age is necessary and feasible.

To assess the efficacy of copy number variation sequencing (CNV-seq) and karyotyping for prenatal detection of chromosomal abnormalities in fetuses with increased nuchal translucency. Amniotic fluid samples were extracted from 205 fetuses with increased nuchal translucency (NT ≥ 2.5 mm), diagnosed by ultrasound between gestational ages of 11 and 13 + 6 weeks. Karyotyping and CNV-seq were performed for detecting chromosomal abnormalities. There are 40 fetuses (19.51%) showing increased NT detected with chromosomal abnormalities in karyotyping, and trisomy 21 was found to be the most common abnormalities. There are 50 fetuses (24.39%) identified with chromosomal abnormalities by CNV-seq. The detection of the applied techniques indicated that CNV-seq revealed higher chromosomal aberrations. The risk of chromosomal abnormalities was significantly increased with NT thickening, from 13.64% in the NT group of 2.5–3.4 mm, 38.64% in the NT group of 3.5–4.4 mm, and to 51.72% in the NT group of over 4.5 mm (P < 0.05). The investigated cases with increased NT with presence of soft markers in ultrasound or high risk in non-invasive prenatal testing presented chromosomal abnormalities in higher rates, comparing with those with isolated NT or low risk (P < 0.05). The results indicated that the risk of chromosomal abnormalities was associated with the NT thickness, detected by karyotype or CNV-seq. The combination application of two analysis was efficient to reveal the possible genetic defects in prenatal diagnosis. The finding suggested that the detection should be considered with ultrasonographic soft markers, and the NT thickness of 2.5–3.4 mm could be a critical value for detecting chromosomal abnormalities to prevent the occurrence of missed diagnosis.

目的:通过分析胎儿半椎体畸形病例的临床资料,为胎儿半椎体畸形的妊娠期管理、产前诊断和遗传咨询提供临床策略.方法:对胎儿超声检查发现胎儿半椎体畸形的孕妇进行系统超声筛查和磁共振(MRI)检查,并采集胎儿组织和夫妻双方外周血进行拷贝数变异检测(CNV-seq)及家系全外显子组检测(Trio-WES).结果:胎儿系统超声筛查和胎儿MRI结果均提示胎儿半椎体、T8-S1椎体排列不整齐,CNV-seq未检出明确致病的基因组拷贝数变异(CNVs),Trio-WES检出TBX6基因c.172dup新发致病性变异,可导致常染色体显性遗传的脊柱肋骨发育不全5型(SCD5),该变异为首次报道.结论:胎儿半椎体畸形应通过胎儿系统性超声、MRI进行检查,并建议进行产前诊断明确遗传病因.相对于常规染色体核型分析和CNVs检测,WES能提高导致胎儿半椎体畸形的基因异常检出率.

This study introduced whole-exome sequencing (WES) in prenatal diagnosis of fetal bowel dilatation to improve the detection outcome when karyotype analysis and copy number variation sequencing (CNV-seq) were uninformative in detecting pathogenic variants. The work reviewed 28 cases diagnosed with fetal bowel dilatation and analyzed the results of karyotype analysis, CNV-seq, and WES. Among the 28 cases, the detection rate in cases with low risk of aneuploidy was 11.54% (3/26), which is lower than 100% (2/2) in cases with high risk of aneuploidy. Ten low-risk aneuploidy cases with isolated fetal bowel dilatation had normal genetic testing results, while the remaining 16 cases with other ultrasound abnormalities were detected for genetic variants at a rate of 18.75% (3/16). The detection rate of gene variation was 3.85% (1/26) by CNV-seq and 7.69% (2/26) by WES. This study suggested that WES could reveal more genetic risk in prenatal diagnosis of fetal bowel dilatation and has value in prenatal diagnosis to reduce birth defects.

目的 探讨基因组拷贝数变异检测（CNV-seq）联合染色体核型分析在评估胎儿心室强光点超声软指标中的临床应用价值。方法 收集2017年6月至2020年12月于华中科技大学同济医学院附属同济医院妇产科胎儿系统超声诊断为心室强光点（EIF）的143例孕妇，羊膜腔穿刺术抽取羊水，羊水标本行染色体核型分析和拷贝数变异检测。结果 143例心室强光点胎儿羊水标本中，核型分析共检出8例异常结果，异常率为5.59%;CNV-seq共检出8例致病性拷贝数变异（pCNV），异常率5.59%，联合检测共发现9例异常结果，异常率6.29%。心室强光点合并其他单个和多个超声软指标组，染色体异常率分别为8.70%和25.00%，明显高于孤立性心室强光点组（0.00%），差异有统计学意义。心室强光点合并非整倍体筛查高风险时，染色体异常率25.00%，明显高于筛查低风险组（4.58%）。结论 孤立性心室强光点不会增加胎儿染色体异常的风险，心室强光点合并其他超声软指标异常或非整倍体筛查高风险时，染色体畸变的风险显著增加。此外，在核型分析的基础上应用CNV-seq能够检测出额外的染色体异常，减少漏诊和出生缺陷的发生。

BACKGROUND:Conventional contact-based electroanatomic mapping is poorly suited for rapid or dynamic ventricular arrhythmias. Whole-chamber charge density (CD) mapping could efficiently characterize complex ventricular tachyarrhythmias and yield insights into their underlying mechanisms.OBJECTIVE:The purpose of this study was to evaluate the feasibility and accuracy of noncontact whole-chamber left ventricular (LV) CD mapping and to characterize CD activation patterns during sinus rhythm, ventricular pacing, and ventricular fibrillation (VF).METHODS:Ischemic scar as defined by CD amplitude thresholds was compared to late gadolinium enhancement criteria on magnetic resonance imaging using an iterative closest point algorithm. Electrograms recorded at sites of tissue contact were compared to the nearest noncontact CD-derived electrograms to calculate signal morphology cross-correlations and time differences. Regions of consistently slow conduction were examined relative to areas of scar and to localized irregular activation (LIA) during VF.RESULTS:Areas under receiver operating characteristic curves (AUCs) of CD-defined dense and total LV scar were 0.92 ± 0.03 and 0.87 ± 0.06, with accuracies of 0.86 ± 0.03 and 0.80 ± 0.05, respectively. Morphology cross-correlation between 8677 contact and corresponding noncontact electrograms was 0.93 ± 0.10, with a mean time difference of 2.5 ± 5.6 ms. Areas of consistently slow conduction tended to occur at scar borders and exhibited spatial agreement with LIA during VF (AUC 0.90 ± 0.02).CONCLUSION:Noncontact LV CD mapping can accurately delineate ischemic scar. CD-derived ventricular electrograms correlate strongly with conventional contact-based electrograms. Regions with consistently slow conduction are often at scar borders and tend to harbor LIA during VF.

Background: It is challenging to make an accurate prenatal diagnosis for congenital anomalies of the kidney and urinary tract (CAKUT) because of its pathologic diversity. This study aims to evaluate the performance of whole-exome sequencing (WES) combined with karyotype analysis and copy number variations (CNVs) in diagnosing high-risk fetal CAKUT.Methods: We conducted a retrospective study on prenatal diagnoses of CAKUT in our hospital from January 2020 to April 2021. The research studied 24 high-risk fetuses with CAKUT who were scanned by ultrasonography at the prenatal diagnosis center of Tongji Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology. The likely pathogenic gene variants were screened for the patients and their parents by multiple approaches, including karyotype analysis, CNVs and WES, and further verified with Sanger sequencing.Results: ①We detected abnormal CNVs in 20.8% (5/24) of the fetuses but only 8.3% (2/24) fetuses had abnormal karyotypes. ②Of the 15 CAKUT fetuses, positive findings (40%) were detected by WES. Of the 9 high-risk fetuses with CAKUT (negative findings in ultrasound scan but with family history), we found abnormal variants (77.8%) through WES.Conclusion: The application of CNVs and WES showed advance in prenatal diagnosis of CAKUT and the pathogenic gene variants were detectable especially for high-risk fetuses with negative ultrasound findings on CAKUT in the preliminary study. The applied strategy could be used to improve the accuracy of prenatal diagnosis for CAKUT in the future.

A novel heterozygous mutation (c.325dup) was identified in EXT1 gene from the proband and the affected family members; this mutation was absent in all the unaffected family members. The identification of the novel frameshift insertion mutation (c.325dup) expands the mutation spectrum of HME, which provides new evidence for HME diagnosis.

BACKGROUND A novel aggregated multiposition noncontact mapping (AMP-NCM) algorithm is proposed to diagnose cardiac arrhythmias. OBJECTIVE The purpose of this study was to computationally determine an accuracy threshold and to compare the accuracy and clinical utility of AMP-NCM to gold standard contact mapping. METHODS In a cellular automata model, the number of catheter positions and chamber coverage were varied to establish accuracy requirements for clinically relevant AMP-NCM. This guided the clinical study protocol. In a prospective cohort of patients with atrial tachycardia (AT), noncontact mapping (NCM) recordings from a single position (SP) and multiple positions were compared to contact mapping with a high-density multipolar catheter using morphology and timing differences of reconstructed signals. Identification of AT mechanisms and ablation targets using both AMP-NCM and contact mapping were randomly evaluated by 5 blinded reviewers. RESULTS AMP-NCM accuracy was asymptotic at 60 catheter positions in computational modeling. Twenty patients (age 65 +/- 12 years; 19 male) with 26 ATs (5 focal, 21 reentrant) were studied. Morphologic correlation of signals derived from AMP-NCM was significantly better than those from SP-NCM compared to contact signals (median 0.93 vs 0.76; P <.001). AMP-NCM generated maps more rapidly than contact mapping (3 +/- 1 minutes vs 13 +/- 6 minutes; P <.001) and correctly diagnosed AT mechanisms in 25 of 26 maps (96%). Overall, 80% of arrhythmia mechanisms were correctly identified using AMP-NCM by blinded reviewers. CONCLUSION Once 60 catheter positions were achieved, AMP-NCM successfully diagnosed mechanisms of AT and identified treatment sites equal to gold standard contact mapping in 3 minutes of procedural time.

To identify the pathogenic gene variation in a Chinese family with Hereditary Multiple Exostoses (HME). By examining blood-sourced DNA and clinical manifestations of the proband and his family members, the whole exome sequencing (WES) and Sanger sequencing were used to detect possibly pathogenic mutations. A novel heterozygous mutation (c.325dup) was identified in exon 1 of the exostosin 1 (EXT1) gene from the proband and the affected family members. And we found this mutation was absent in all the unaffected family members. This c.325dup mutation is in the exon 1 domain of the EXT1 gene and the change of p.C109Lfs*80 cause the early termination of protein translation. The identification of the novel frameshift insertion mutation (c.325dup) expands the mutation spectrum of HME, which provides new evidence for HME diagnosis.

目的:利用全外显子测序(WES)技术分析胎儿肠管增宽的致病性突变,为产前诊断及遗传咨询提供帮助.方法:对3例肠管增宽的胎儿进行染色体核型分析、拷贝数变异(CNV)及WES检测.结果:3例胎儿染色体核型分析结果均正常,1例胎儿CNV检出15号染色体微重复,3例胎儿WES结果均发现与肠管增宽相关的基因突变.病例1为GDNF基因c.329G>A杂合变异,遗传自其父亲,可引起先天性巨结肠症3型,该病为常染色体显性遗传,并具有低外显率的特点;病例2为NOTCH2基因的c.1310A>C新发变异,与Alagille综合征2型、Hajdu-Cheney综合征相关,均为常染色体显性遗传;病例3为SLC26A3基因复合杂合突变,包括来自父亲的c.1427del突变和来自母亲的c.269_270 dup突变,可导致常染色体隐性遗传的先天性失氯性腹泻1型.经Sanger验证病例1为临床意义未明突变,根据既往病史判断该突变可能致病,病例2及病例3均为致病性突变.结论:WES技术对于分析胎儿肠管增宽的遗传变异具有重要的诊断价值.

Conventional activation mapping is achieved in stable rhythms by annotating local activation times compared to a reference EGM. Efficiently mapping quasi stable rhythms such as short bursts of AT during SR or AF may be of interest, but traditional mapping approaches are unable to adapt to such transitory events. Real-time, auto-identification of unique rhythms during transient conditions can facilitate adaptive mapping of multiple rhythms during a single acquisition.

Non-contact (NC) mapping efficiently acquires global activation wavefronts and is potentially more accurate than contact (C) mapping due to elimination of C-related artifact. Common areas of conduction velocity (CV) deceleration during sinus and paced rhythms have been used to characterize the atrial substrate, however, such analysis with C mapping is time-consuming.

Non-contact charge density (NCCD) mapping (Acutus, Carlsbad, CA) can be used for verification of pulmonary vein isolation (PVI) which is conventionally verified using contact mapping to demonstrate loss of signal and/or pacing to confirm exit/entrance block.

The proband is a five-year-old boy diagnosed with Duchenne muscular dystrophy (DMD) by clinical manifestations and laboratory examination, but clinical phenotype of his parents is normal. In the study, his mother had a second pregnancy, and they went to obstetrics for genetic counseling to make informed reproductive choices.

The Duchenne Muscular Dystrophy (DMD) gene variants are associated with the disease phenotypes. The pathogenic mutation, c.2293-1G>C, was detected in DMD gene in the proband and the fetus, which has not been reported in the literature.The minigene expression in vitro confirmed that c.2293-1G>C is responsible of aberrant splicing.