Hailong Huang

Objectives: To characterize the clinical and genetic features of Phelan–McDermid syndrome (PMS) in a Chinese prenatal and postnatal cohort, explore genotype–phenotype correlations, and provide evidence to support prenatal genetic counseling. Methods: G-banded karyotyping, single-nucleotide polymorphism arrays (SNP arrays), copy number variation sequencing, and trio whole-exome sequencing were used for genetic diagnosis. Fifteen patients with PMS (11 prenatal and 4 postnatal) were retrospectively enrolled. Genomic visualization, protein structural prediction, and phenotypic heatmap analyses were conducted to analyze genotype–phenotype associations. Results: The prenatal and postnatal detection rates of PMS were 0.061% and 0.49%, respectively. Only 13.3% of cases were detected by karyotyping, whereas 14 cases were confirmed by SNP array analysis. All 22q13 deletions (69.4 kb–8.5 Mb) involved SHANK3, and all copy number variants were de novo. A novel SHANK3 frameshift variant, c.3513_3514delCC, was identified and predicted to result in protein truncation leading to intellectual disability. Prenatal cases mainly presented non-specific ultrasound anomalies (63.6%, 7/11), with fetal growth restriction, renal malformations, and cardiovascular defects being the most frequent findings. A notable 36.4% (4/11) of prenatal cases were complicated by missed abortion, whereas postnatal patients showed predominant neurodevelopmental impairments. In our cohort, larger deletions were observed in cases presenting with more extensive multisystem involvement, a pattern that mirrors findings from recent large-cohort studies. central nervous system abnormalities were predominantly linked to SHANK3 haploinsufficiency, while multi-system malformations seemed to correlate with the cumulative loss of multiple genes across the 22q13 region, in keeping with current hypotheses on contiguous gene effects. Conclusions: PMS presents obvious prenatal–postnatal phenotypic heterogeneity. SHANK3 is the core pathogenic gene, and deletion may influence the extent of multisystem involvement. Combined genetic testing can improve the accuracy of prenatal diagnosis and counseling for PMS.

Copy number variation (CNV) is defined as a > 1-kb-long DNA fragment copy number increase or decrease, with fetal CNV judgment and genetic counseling posing difficulties. We tested and retrospectively analyzed fetuses using chromosomal microarray analysis (CMA), 169 with abnormal CNV being completely verified and followed up for pregnancy and postnatal outcomes. Among the 169 fetuses, 103 and 66 exhibited inherited and de novo CNV. Of the former, 50, 34, and 19 displayed hereditary pathogenic CNV (P-CNV) [likely pathogenic CNV (LP-CNV)], hereditary variants of uncertain significance (VUS), and hereditary benign (likely benign) CNV, respectively. Of the 50 fetuses with hereditary P-CNV (LP-CNV), 16, one, and two were terminated, died two days after birth, and retained abnormal postnatal phenotypes, respectively. Of the 34 fetuses with hereditary VUS CNV, 5 were terminated. Of the 19 fetuses with hereditary benign (likely benign) CNV, one was terminated and another died of heart disease after birth. Of the 66 fetuses with de novo CNV, 50, 13, and three had P-CNV (LP-CNV), VUS, and benign (likely benign) CNV, respectively. Of the 50 de novo P-CNV (LP-CNV) fetuses, 34 were terminated, one died after birth, and another was born with congenital heart defects. Of the 13 de novo VUS CNV fetuses, six were terminated. Finally, of the three de novo benign (likely benign) CNV fetuses, two were terminated and one displayed developmental delays after birth. This study highlights that CMA detects small, mostly inherited, P-CNV. Even P-CNV could yield favorable outcomes, while benign results do not preclude adverse events, especially with ultrasound anomalies. Evolving genomic data would refine CNV interpretation. Our findings enhance prenatal counseling by balancing the utility of inheritance information with prognostic uncertainty, supporting a personalized approach to genetic risk communication.

Spontaneous single intrauterine fetal demise (SSIUFD) in twin pregnancies is a special condition of stillbirth. When SSIUFD occurs, clinical attention understandably shifts to the surviving co‑twin. This study aimed to investigate the genetic profiles and perinatal outcomes of surviving co-twins following SSIUFD. This retrospective study reviewed 88 twin pregnancies complicated by spontaneous SSIUFD that underwent invasive prenatal testing between January 2015 and December 2024. Cases of elective fetal reduction and those with unknown chorionicity were not included. The cohort included 54 dichorionic diamniotic (DCDA) and 34 monochorionic diamniotic (MCDA) pregnancies. According to gestational age at the single twin demise, they were classified into the first-trimester SSIUFD (≤ 12 weeks’ gestation) and the second-trimester SSIUFD group (> 12 weeks’ gestation). Genetic evaluation was performed using karyotyping and/or single nucleotide polymorphism (SNP) array analysis on samples of amniotic fluid (n = 85) and cord blood (n = 3). The prognosis of the surviving co-twins including prenatal ultrasound monitoring and delivery outcome were collected and analyzed according to the gestational age at fetal demise and chorionicity. Of 88 cases, 24 (27.3%) cases underwent invasive prenatal diagnosis because of SSIUFD alone, while the remaining 64 (72.7%) cases had additional clinical indications. Among the 24 cases tested for SSIUFD alone, no clinically significant chromosomal abnormalities were detected (0/24, 0%). A low incidence of chromosomal abnormalities (1.1%, 1/88) was observed, with one case of trisomy 21 in a MCDA pregnancies, both twins exhibiting concordant ultrasound anomalies. SNP array analysis revealed no pathogenic submicroscopic variants in 68 analyzed cases. Among 83 cases with follow-up information available, MCDA pregnancies showed significantly higher termination rates than DCDA (19.4% vs. 1.9%, p < 0.05), primarily due to severe anomalies or subsequent fetal demise followed SSIUFD. Overall, term deliveries occurred in 81.9% and preterm birth (PTB) in 9.6%, with comparable PTB rates (p > 0.05) between DCDA (13.5%) and MCDA (6.5%) groups, or between the first- (6.0%) and the second-trimester SSIUFD group (15.2%). Our findings do not provide evidence for an increased detection rate of chromosomal abnormalities in surviving co-twins after SSIUFD, particularly in surviving co-twins without other indications. Surviving co-twins from MCDA pregnancies are associated with a higher risk of adverse outcomes, including termination due to fetal malformations and subsequent fetal demise. This highlights the importance of close ultrasound monitoring in these cases.

Posttranslational modifications (PTMs) act as rapid, reversible switches that reshape protein activity, stability, and interactome, thereby governing virtually every physiological cue from signal transduction to epigenetic memory. Mass spectrometry‐based proteomics has considerably extended our knowledge about the occurrence and dynamics of PTMs. Pinpointing disease‐ or physiology‐specific PTM sites remains an open challenge. As a biological process with well‐defined stage‐specific markers and a precise endpoint, erythropoiesis is orchestrated by the complex interplay of multiple PTM‐regulatory networks, making it an ideal model for dissecting the spatiotemporal dynamics, quantitative thresholds, and crosstalk of PTMs. This review delineates the applications, detection, and prediction technologies of PTMs, with an emphasis on the mechanisms of phosphorylation, ubiquitination, methylation, SUMOylation, glycosylation, and acetylation in both physiological and pathological processes. Dissecting PTM circuitry driving erythroid specification and maturation, we show how its perturbation triggers disease, clarifying PTM roles. Additionally, we have investigated the progress made in the clinical translation and drug development of the PTMs field, emphasizing the potential of PTMs in the field of precision medicine as well as the existing challenges. This review aims to provide new insights and perspectives for the study of PTMs.

Background/Objectives: This study assesses the genetic basis of fetal congenital heart disease (CHD), which exhibits a complex etiology, by using chromosomal microarray analysis (CMA); it also elucidates perinatal outcomes and postnatal development to support prenatal diagnosis and genetic counseling. Methods: Pregnant women (n = 1195) who were diagnosed with fetal CHD based on echocardiography were selected along with those having an interventional prenatal diagnosis, all of whom underwent CMA. Depending on the gestational age, amniotic fluid or umbilical cord blood samples were collected. Patients were included if they were diagnosed with fetal CHD based on echocardiography. Those who could not consent to amniocentesis or umbilical vein puncture or who had contraindications for amniocentesis or umbilical vein puncture were excluded. Patients were studied until May 2025. Results: Of the 1195 fetuses with CHD, 140 had pathogenic copy number variation (pCNV). The pCNV detection rate in cases with a single CHD was 3.17%, whereas it was 13.51% in the group with multiple CHDs. The detection rate for pCNVs in patients with extracardiac abnormalities was 28.62%. The fetal and postnatal mortality rates were highest for fetuses with multiple CHDs. The survival rate was highest for fetuses with a single CHD. Early detection of CHD and timely genetic testing can inform clinical management of CHD-affected pregnancies; however, larger prospective studies are needed to establish their impact on perinatal outcomes. Conclusions: CMA provides valuable information for genetic counselling, as it identifies pathogenic variants associated with CHD. However, prognostic predictions should consider multiple clinical factors.

β-Thalassemia is a chronic hemolytic anemia caused by mutations in the β-globin gene. Increasing studies have shown that circular RNA (circRNA) plays a key role in abnormal hematopoiesis. Circ_0001428 is formed by circularization of the beta-mannosidase gene. However, little is known about the biological function of circ_0001428 in β-thalassemia. Circ_0001428 was selected from two circRNA microarrays (GSE241141 and GSE196682) of β-thalassemia, and expression levels of circ_0001428, miR-32-3p, miR-325-3p, γ-globin, activating transcription factor 4 (ATF4) and B-cell lymphoma/leukemia 11 A (BCL11A) were validated in 30 patients with β-thalassemia and 30 healthy controls. Cell proliferation, apoptosis, cell cycle, and erythroid differentiation were evaluated using stable knockdown or overexpression of circ_0001428. Fluorescence in situ hybridization, dual-luciferase reporter, and Western blots were performed to confirm interactions between circ_0001428, miR-32-3p/miR-325-3p, and ATF4. Circ_0001428 was downregulated in β-thalassemia patients compared to controls, promoted cell proliferation, decreased cell apoptosis, and inhibited terminal erythroid differentiation and γ-globin production. Mechanistically, circ_0001428 functions as a sponge for miR-32-3p and miR-325-3p to promote ATF4 expression, thereby activating BCL11A expression. Additionally, rescue experiments confirmed that circ_0001428 regulated erythropoiesis and γ-globin expression via promoting the ATF4-BCL11A axis by binding with miR-32-3p/miR-325-3p. Our findings reveal, for the first time, the important role of circ_0001428 in regulating erythropoiesis and γ-globin expression in β-thalassemia. The circ_0001428/miR-32-3p and miR-325-3p/ATF4/BCL11A/γ-globin signaling pathway may be a potential therapeutic target in β-thalassemia, especially for fetal hemoglobin modulation.

Background: Bisphenol A (BPA), a typical endocrine-disrupting chemical, is implicated in the pathogenesis of Polycystic Ovary Syndrome (PCOS); however, the underlying molecular mechanisms and pathophysiological processes remain unclear. This study aims to decipher molecular interactions between BPA and PCOS-related genetic networks, and to determine the combinatorial impacts of environmental pollutants on PCOS progression. Methods: We first identified overlapping genes associated with bisphenol A (BPA) exposure and polycystic ovary syndrome (PCOS) using the Comparative Toxicogenomics Database (CTD). Differentially expressed genes (DEGs) were extracted from three Gene Expression Omnibus (GEO) datasets, while oxidative stress- and apoptosis-related genes were retrieved from the GeneCards database. Subsequently, a series of in silico analyses were performed, including protein-protein interaction (PPI) network construction, functional enrichment profiling, Gene Set Enrichment Analysis (GSEA), immune infiltration evaluation, nomogram development, CB-DOCK molecular docking, and single-cell RNA-seq analysis of the mouse ovarian dataset GSE268919 (DHEA-induced PCOS-like model) to provide cell-type-resolved evidence. Finally, in vitro validation was conducted using primary granulosa cells from PCOS patients and healthy controls, as well as KGN cells, to assess hub gene expression. Functional evaluations were carried out via CCK-8 assay, flow cytometry, quantitative polymerase chain reaction (qPCR), and Western blotting. Results: We identified 139 hub genes between BPA exposure and PCOS, with enrichment in hormone metabolism, ovarian steroidogenesis, and reproductive signaling pathways—among which the apoptotic pathway was prominently associated with these hub genes, indicating BPA exerts a profound impact on cell survival in PCOS. Five hub genes (PTAFR, RACGAP1, CYP19A1, FSHR, DMD) were pinpointed, and a nomogram integrating these genes showed robust PCOS predictive accuracy. Single-gene GSEA further linked the hub genes to immune modulation, inflammation, and cell apoptosis—validating their functional relevance to apoptotic processes in PCOS. Immune cell infiltration analysis revealed discrepancies between PCOS and control groups, with hub genes correlating with specific immune subsets (e.g., pro-inflammatory cells) that may exacerbate apoptotic signaling in ovarian tissues. Molecular docking demonstrated strong binding affinity between BPA and the protein products of hub genes, suggesting direct BPA-mediated interference with their roles in regulating cell apoptosis. In the mouse ovarian scRNA-seq dataset (GSE268919), we observed cell-type-specific dysregulation of Cyp19a1 and Dmd (mouse gene symbols), with stress/apoptosis signatures enriched in specific ovarian cell populations, thereby providing supportive cell-type localization for the hub-gene–associated phenotypes. In vitro validation confirmed dysregulated expression of hub genes in PCOS primary granulosa cells; BPA treatment dose-dependently regulated hub gene expression, inhibited KGN cell proliferation, and significantly induced granulosa cell apoptosis. Conclusion: BPA exposure disrupts granulosa cell survival in PCOS by driving apoptosis-related molecular reprogramming through key gene regulation, thereby elucidating mechanistic links between environmental pollutants and PCOS progression and highlighting potential molecular targets for intervention.

Polycystic ovary syndrome (PCOS) is increasingly viewed as an environmentally influenced, epigenetically mediated reproductive–metabolic disorder. Bi

Introduction: Claes-Jensen syndrome is a rare X-linked syndromic neurodevelopmental disorder by pathogenic variants in lysine specific demethylase 5C ( KDM5C ), a lysine-specific histone demethylase. Methods: In this study, clinical evaluations were conducted in affected individuals and carrier females. X-chromosome inactivation (XCI) assays were performed to assess genotype—phenotype correlations. Functional studies evaluated variant effects on RNA transcription, protein expression, and stability. Zebrafish models were used for in vivo validation. RNA sequencing with KEGG and GO analyses identified dysregulated genes and pathways, further confirmed in zebrafish. Results: Two novel KDM5C variants NM_004187.5:c.3019del and NM_004187.5:c.782-2A>T were identified in unrelated families with X-linked ID. Affected males presented with short stature, microcephaly, language delay, and intellectual disability, while carrier females showed milder features including learning difficulties and short stature. Skewed XCI in some carriers suggested a role in phenotypic variability. Both variants impair RNA transcription, protein expression and stability. Zebrafish models recapitulated neurodevelopmental and behavioral abnormalities. Transcriptomic analyses revealed disrupted antiviral and interferon-related signaling, implicating aberrant immune activation. Pharmacologic inhibition of the Toll-like receptor pathway ameliorated mutant phenotypes, highlighting neuroinflammation as a potential therapeutic target for KDM5C -related disorders. Conclusion: These findings expand the mutational spectrum of KDM5C -associated ID and uncover a novel pathogenic mechanism between KDM5C dysfunction, protein instability, and dysregulated inflammatory signaling.