Jiangxi Provincial Children's Hospital

Necrotizing enterocolitis (NEC) is a severe gastrointestinal disease affecting premature infants, yet its precise pathogenic mechanisms remain unclear. While immunothrombosis, an interplay between inflammation and coagulation, is well recognized in systemic inflammatory diseases, its role in NEC pathogenesis and local intestinal regulation has been incompletely characterized. Here, we combine multi-omics profiling, histopathological analyses, and functional studies using clinical NEC samples and neonatal mouse models. We demonstrate prominent intestinal thromboinflammation in NEC, histologically characterized by immunothrombosis with infiltrating CD177⁺ neutrophils, activated platelets, and fibrin deposition. We show that extensive formation of CD177⁺ neutrophil-platelet aggregates (NPAs) amplifies intestinal injury through neutrophil extracellular trap (NET)-mediated thromboinflammation. Blocking CD177⁺ NPAs or inhibiting NET formation significantly alleviates NEC severity in mouse models. Clinically, elevated CD177⁺ NPA levels correlate with NEC severity and increased thrombotic biomarkers, highlighting their potential as diagnostic and prognostic biomarkers. Importantly, prophylactic administration of low-molecular-weight heparin (LMWH) disrupts NPA formation, reduces NET burden, and improves survival in experimental NEC. Collectively, these findings establish CD177⁺ NPAs as central drivers of NEC pathogenesis, support a thromboinflammatory framework for NEC, and highlight LMWH therapy and CD177⁺ NPA monitoring as possible clinically translatable strategies for neonatal care.

Abstract Introduction: Despite significant improvements in the overall survival of Wilms tumor (WT), a subset of patients still experiences poor outcomes. Programmed cell death (PCD) pathways are pivotal in cancer progression. A deeper understanding of their roles in WT is crucial for harnessing these mechanisms to optimize risk stratification. Methods: Key tumor-associated genes were identified through limma differential analysis and WGCNA, and subsequently integrated with 12 distinct PCD patterns. TAGET-WT transcriptomic data was divided into training and validation sets to construct and validate a risk stratification model. It was subsequently integrated with clinical information to build a comprehensive prediction model. Immune infiltration and drug sensitivity analyses were performed. External validation was performed using scRNA-seq data from the GSE200256 dataset. Results: Key tumor-associated genes were enriched in multiple PCD pathways. The risk stratification model was constructed using 4 genes selected via the machine learning algorithm, stratifying the cohort into high- and low-risk groups. In the overall WT cohort, the high-risk group exhibited poorer prognosis, with 1-, 3-, and 5-year AUC values of 0.820, 0.721, and 0.728, respectively. DCA demonstrated the superior predictive accuracy of the comprehensive prediction model. The high-risk group showed lower infiltration of TH17 cells and increased sensitivity to paclitaxel and sorafenib. Finally, the expression landscape of hub genes was validated in the single-cell dataset. Conclusion: These results highlight a critical role for PCD genes in the progression and immune regulation of WT. Targeting these genes offers a promising avenue for improving clinical management of patients identified as high-risk.

To enhance clinicians' understanding of Sifrim-Hitz-Weiss syndrome (SIHIWES), this study investigated the clinical phenotypes, genetic characteristics, and response to growth hormone therapy in a patient. A case of a patient with global developmental delay and distinctive facial features is presented. To identify the underlying etiology, peripheral blood samples were collected from the patient and both parents. Whole-exome sequencing (WES) and genomic copy number variation (CNV) analysis were conducted. Candidate variants were confirmed by Sanger sequencing within the family, and bioinformatics tools assessed their pathogenicity. WES identified a de novo heterozygous variant, c.3547C>T (p.Arg1183Cys), in CHD4. Both parents were wild-type at this locus, and CNV analysis revealed no pathogenic variants. Based on ACMG guidelines, this variant is classified as pathogenic. Combining clinical phenotypes with genetic findings, this study confirmed the diagnosis of Sifrim-Hitz-Weiss syndrome. Protein structure modeling indicated that the CHD4 protein harbors an Arg1183Cys variant in the α-helix. The cysteine side chain, containing a sulfur atom, may introduce new chemical interactions, potentially altering CHD4 protein function. Short-term growth-promoting effects were observed with recombinant human growth hormone (rhGH) therapy, providing new insights for managing this disorder. Long-term efficacy and safety should be evaluated in larger studies.

This study aims to explore the functions and mechanisms underlying the involvement of 14-3-3β in esophageal squamous cell carcinoma (ESCC). Western blot and quantitative real-time PCR (RT-PCR) were utilized to evaluate the levels of 14-3-3β in SHEE, TE-1, and KYSE-150 cells. Following infection with YWHAB-RNAi lentivirus and puromycin screening, the reduction of 14-3-3β expression in KYSE-150 cells was confirmed via western blot and RT-PCR. Subsequently, wound healing, CCK8, and colony-formation assays were performed to assess cell migration and proliferation in KYSE-150 cells with stable low levels of 14-3-3β. Furthermore, western blot analysis was conducted to examine the expression of relevant proteins, elucidating the potential molecular mechanisms underlying 14-3-3β involvement in ESCC progression and metastasis. The expression of 14-3-3β was significantly elevated in KYSE-150 cells compared to SHEE cells. Upon infection in KYSE-150 cells, the expression of 14-3-3β was diminished, resulting in suppressed cell migration and colony formation, although no significant changes were observed in the CCK8 assays. Furthermore, in KYSE-150 cells exhibiting reduced 14-3-3β expression, the activity of phosphorylated AKT (p-AKT) was significantly decreased, while no notable difference was detected in AKT protein levels. Additionally, there were no significant alterations observed in apoptosis-related proteins, including BCL2, BAX, Caspase3, and activated Caspase9. In KYSE-150 cells infected with YWHAB-RNAi lentivirus, the downregulation of 14-3-3β expression resulted in significant reductions in both cell migration and proliferation. Furthermore, the findings demonstrated that 14-3-3β may regulate the biological activities of ESCC cells via modulation of the PI3K/AKT signaling pathway.

ObjectiveThis study aimed to explore the role of miR-194 in the post-transcriptional regulation of PTPN12 and its impact on age-related thymic atrophy.MethodsmiR-194-5p and PTPN12 expression levels in thymus tissues from young and aged mice were assessed using quantitative PCR and western blot analysis. An aged-mouse model was established to overexpress varying levels of miR-194-5p. Expression levels of PTPN12, AKT, BAX, p50, and p65 in thymic epithelial cells were analyzed via quantitative PCR and western blot. Flow cytometry was used to evaluate TEC cell cycle progression, proliferation, and apoptosis. CD4+/CD8+ lymphocytes proportions in the spleen and peripheral blood were quantified, and structural changes in the thymus were examined using hematoxylin–eosin staining.ResultsmiR-194 expression peaked in the thymus of middle-aged (6–7 months) C57BL/6 mice, while PTPN12 expression was highest in the young (1–2 months) mice and declined with age. TEC survival was enhanced in the miR-194-5p overexpression group, as indicated by cell cycle and apoptosis analyses. Additionally, CD4+ and CD8+ T cell counts and proportions in the spleen and peripheral blood were reduced in the miR-194-5p overexpression group. Compared to controls, the high-dose miR-194-5p group exhibited significantly increased p65 and p50 expression, while PTPN12 and BAX levels were markedly reduced, but AKT level was markedly increased.ConclusionThis study demonstrated that miR-194-5p modulates thymic immunological function in aged mice, highlighting its potential role in age-related thymic atrophy.

Postoperative cognitive dysfunction (POCD) is a common complication of coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB), largely associated with cerebral hemodynamic disturbances. Carotid duplex ultrasonography–derived parameters, including peak systolic velocity (PSV-ICA) and end-diastolic velocity (EDV-ICA), provide reliable, noninvasive indicators of cerebral blood flow. This study evaluated whether intraoperative nicardipine infusion improves carotid blood flow and mitigates postoperative cognitive decline. In this randomized, double-blind trial, 64 elderly patients undergoing CABG were assigned to receive either continuous low-dose nicardipine (0.2–0.5 µg/kg·min) during CPB (nicardipine group, n = 32) or saline (control group, n = 32). The co-primary endpoints were intraoperative PSV-ICA and EDV-ICA at four predefined time points (T1: pre-anesthesia; T2: surgery initiation; T3: during CPB; T4: post-CPB). Secondary endpoints included regional cerebral oxygen saturation (rScO₂), plasma neuron-specific enolase (NSE), Mini-Mental State Examination (MMSE) scores, vasopressor use, ventilation time, ICU stay, and hospitalization length. Baseline and intraoperative characteristics were comparable. Both groups showed reduced carotid flow velocities during CPB, but recovery of PSV-ICA at the end of CPB was significantly greater with nicardipine (63.7 ± 15.2 vs. 54.3 ± 13.9 cm/s; P = 0.025). EDV-ICA exhibited a similar trend without significant between-group differences. Secondary outcomes (MMSE, NSE) did not differ, and no adverse events occurred. Continuous low-dose nicardipine during CPB significantly improved the recovery of PSV-ICA without affecting systemic hemodynamics or increasing adverse events. These findings suggest a potential intraoperative neuroprotective role of nicardipine, warranting confirmation in larger trials with long-term cognitive outcomes.

Wilms tumor is a highly heterogeneous tumor, and patients with high-risk Wilms tumor still do not have a good prognosis. DNA instability caused by abnormal genes in the base excision repair (BER) pathway is closely related to cancer. However, the important role of BER pathway gene variants in Wilms tumor remains largely unknown. We enrolled 621 patients with Wilms tumor and 1737 controls from eight hospitals. We calculated odds ratios and 95% confidence intervals to evaluate the association strength. Stratification analysis was performed to further evaluate the associations of significant polymorphisms with the risk of Wilms tumor in different subgroups. False-positive report probability analysis was adopted to evaluate the robustness of the positive results. The associations of polymorphisms with gene expression were analyzed via eQTLs from the GTEx database. We found that the FEN1 rs174538 GG, FEN1 rs4246215 GG, APEX1 rs3136817 CC, and XRCC1 rs25487 TT genotypes were associated with an increased risk of Wilms tumor, whereas the FEN1 rs4246215 TG/GG genotype was associated with a decreased risk. Stratification analysis revealed that significant polymorphisms remained associated with the risk of Wilms tumor in some subgroups. False-positive report probability analysis also revealed that some positive results were more robust. The eQTL results revealed that all four polymorphisms were associated with alterations in the expression of host or nearby genes. FEN1 rs174538 A>G, FEN1 rs4246215 T>G, APEX1 rs3136817 T>C, and XRCC1 rs25487 C>T are associated with Wilms tumor susceptibility, which provides potential molecular markers for the early diagnosis of Wilms tumor.

Breakthrough urinary tract infection (BT-UTI) among children with vesicoureteral reflux (VUR) receiving continuous antibiotic prophylaxis (CAP) remains a clinical challenge with heterogeneous risk profiles and no validated prediction tools. We aimed to identify robust risk factors and develop an evidence-based predictive score for BT-UTI. We conducted a systematic review and meta-analysis following PRISMA and Cochrane guidance, searching PubMed, Web of Science, and Embase for cohort studies. Pooled effect estimates were calculated using appropriate fixed- or random-effects models; a scoring system was derived by natural logarithm transformation of pooled estimates. External validation used a cohort of 158 children with VUR receiving CAP at Children's Hospital of Chongqing Medical University (2021-2024). Twenty-four cohort studies (n = 3,264) were included; pooled BT-UTI incidence was 30.5%. From 26 candidate factors, eight stable predictors were retained and incorporated into the score: age < 1year (OR 2.04), bilateral reflux (OR 1.81), high-grade reflux IV-V (OR 2.65), symptomatic UTI (OR 2.74), history of recurrent UTI (OR 2.17), bladder and bowel dysfunction (BBD; OR 1.81), increased ureteral diameter ratio (UDR; MD 0.139), and DMSA renal scar formation (OR 4.40). External validation showed AUC 0.88 (95% CI 0.82-0.93); at a cutoff of 17 points accuracy was 0.84, and sensitivity 0.93. Calibration and decision-curve analyses indicated good agreement and a positive net clinical benefit. We developed and externally validated an eight-item BT-UTI risk score for children with VUR on CAP that demonstrates high discrimination and clinical utility for individualized risk stratification and prevention planning and implementation guidance available.