OBJECTIVE:To evaluate the clinical value and efficacy of colonic segment reversal in children with short bowel syndrome (SBS). METHODS:A retrospective review was conducted on 11 children with SBS who underwent colonic segment reversal at our center between January 2016 and October 2025. Postoperative intestinal transit time, weaning rate from parenteral nutrition (PN), and growth parameters (weight-for-age z-score, WAZ), complications, and frequency of small intestinal bacterial overgrowth (SIBO) were evaluated. RESULTS:Eleven children (8 males, 3 females) underwent colonic reversal. The overall survival rate was 100% (11/11), and the PN weaning rate was 63.6% (7/11). The median time to PN weaning was 3 months. Postoperative intestinal transit time was prolonged compared with preoperative values (data available for 3 patients). The median preoperative WAZ was -1.85, which improved to -0.87 at 6 months postoperatively. Complications included one case of reversible intestinal obstruction and one case of anastomotic ulcer. The frequency of small intestinal bacterial overgrowth did not increase significantly. CONCLUSION:Colonic segment reversal is a safe and effective surgical option for selected children with SBS, particularly when residual small bowel length is extremely limited. Surgical strategies should be individualized, taking into account residual bowel length, motility, and anatomical conditions.

Omphalocele is a central abdominal wall defect that can be readily diagnosed at birth. Cantrell syndrome (CS) is an exceedingly rare disorder characterized by abnormalities involving the abdominal wall, sternum, diaphragm, pericardium, and heart, with a poor prognosis. The processes of diagnosis and treatment may pose challenges due to the variable presentation and severity of the condition. Here, we report our experiences in managing a neonate with Cantrell syndrome. A female neonate presented with an abdominal mass, initially diagnosed as an omphalocele accompanied by congenital heart disease based on its appearance, abdominal computed tomography, and echocardiography. The final diagnosis of Cantrell syndrome was confirmed during the omphalocele repair operation, where a pulsatile heart and a diaphragm defect were observed in the hernial sac. During the abdominal reconstruction procedure, an episode of pulseless electrical activity occurred, requiring immediate intracardiac compressions, which successfully restored cardiac activity. She subsequently underwent two cardiovascular surgeries to repair a ventricular septal defect (VSD), an atrial septal defect (ASD), and a patent ductus arteriosus (PDA) a few days later. However, she experienced frequent episodes of supraventricular tachycardia (SVT) postoperatively. During the 20 months of follow-up, the patient exhibited good recovery, although SVT persisted, necessitating long-term administration of antiarrhythmic medication. Concomitant anomalies should always be considered when diagnosing an omphalocele. Although rare, Cantrell syndrome ought to be suspected in cases exhibiting an epigastric omphalocele. Further detailed examinations and comprehensive assessments are essential for establishing a definitive diagnosis and developing an appropriate treatment plan. The involvement of a multidisciplinary team is imperative.

Since their discovery, Z-nucleic acids (Z-NAs), which adopt a left-handed double helical conformation, have puzzled researchers regarding their physiological functions. These unusual nucleic acids are recognized by proteins containing Zα domains, particularly Z-DNA binding protein 1 (ZBP1) and adenosine deaminase acting on RNA 1 (ADAR1). Utilizing mouse genetics with knockout models and site-specific Zα domain mutations, scientists have revealed that Z-NAs serve as critical regulators of programmed cell death, inflammation, antiviral immunity, and anti-tumour responses. This review systematically examines mechanistic insights from Zbp1- and Adar1-mutant models, illuminating how Z-NAs play a dual role as essential triggers of host defence and as potential drivers of autoinflammatory diseases.

Objective: Intraoperative hypotension is associated with adverse postoperative outcomes, yet evidence in the neonatal population is limited. This study aimed to investigate the incidence, risk factors, and prognostic significance of intraoperative hypotension in this population. Methods: This single-center retrospective study included 1712 neonates undergoing non-cardiac surgery from 2020 to 2025 at a tertiary children’s hospital in China. Intraoperative hypotension was defined as a reduction in mean arterial pressure of more than 20% from the preoperative baseline and lasting for more than 5 min. Multivariate logistic regression identified independent risk factors. Results: The incidence of intraoperative hypotension was 59.8% (1024/1712). Independent risk factors included American Society of Anesthesiologists Physical Status Classification > Ⅲ (OR = 1.357, 95% CI: 1.079–1.706, P = 0.009), emergency surgery (OR = 1.323, 95% CI: 1.054–1.660, P = 0.016), surgery duration > 60 min (OR = 1.457, 95% CI: 1.162–1.826, P = 0.001), thoracic surgery (OR = 1.727, 95% CI: 1.053–2.831, P = 0.030), preoperative congenital heart disease (OR = 1.379, 95% CI: 1.117–1.702, P = 0.003), and preoperative acidosis (OR = 1.349, 95% CI: 1.036–1.757, P = 0.026). Higher gestational age (OR = 0.936, 95% CI: 0.891–0.983, P = 0.008) and neurosurgery (OR = 0.566, 95% CI: 0.385–0.832, P = 0.004) were protective factors. Intraoperative hypotension was associated with prolonged postoperative recovery and increased acute kidney injury and intraventricular hemorrhage (all P < 0.05). Conclusions: Intraoperative hypotension is highly prevalent in neonatal non-cardiac surgery and was associated with short-term outcomes. Identified risk and protective factors may guide perioperative hemodynamic optimization in this vulnerable population.

Necrotizing enterocolitis (NEC) remains a leading cause of morbidity and mortality among preterm neonates. However, the timely diagnosis of NEC is hindered by the lack of reliable biomarkers. A key pathogenic feature of NEC is the overproduction of hypochlorous acid (HClO) by activated neutrophils and macrophages via the MPO-H2O2-Cl-system. Therefore, the development of robust HClO detection tools is crucial for the early diagnosis of NEC. In this study, a novel near-infrared (NIR) fluorescent probe (Cy-1) was developed to monitor HClO, with the aim of enabling early NEC diagnosis. Cy-1 displayed high sensitivity and selectivity toward HClO with a rapid response (40 s), and was capable of detecting both exogenous and endogenous HClO in living cells. Using this probe, we successfully tracked dynamic changes in HClO levels during the cisplatin-induced apoptosis. Furthermore, in vivo fluorescence imaging showed significantly increased fluorescence signals in NEC model mice after Cy-1 administration. This work demonstrated the great potential of Cy-1 as a powerful tool for investigating HClO-associated apoptotic processes, as well as a promising candidate for the early clinical diagnosis of NEC.

BACKGROUND:The diagnosis and surgical prediction of necrotizing enterocolitis (NEC) remain challenging. Our goal is to develop an interpretable multimodal artificial intelligence model to assist these key clinical decisions. METHODS:This retrospective study included 484 neonates (242 with NEC, 242 without NEC). We developed a dual Swin Transformer integrating abdominal X-rays (2D branch) and laboratory parameters (1D branch) via late fusion. The model was refined using an external data domain adaptation strategy (n = 50) and evaluated on independent internal and external test sets. The interpretability of the model was evaluated by Grad-CAM and SHAP. RESULTS:The optimized multimodal model showed high performance on the internal test set, achieving AUCs of 0.915 for NEC diagnosis and 0.920 for surgical prediction. On the independent external test set, it achieved AUCs of 0.903 (diagnosis) and 0.894 (surgical prediction), significantly outperforming baseline models. Interpretability analyses highlighted clinically relevant features, including intestinal pneumatosis and specific inflammatory markers (such as C-reactive protein) as key predictive factors. CONCLUSIONS:The dual Swin Transformer provides an accurate, interpretable, and adaptable multimodal tool that integrates radiographic and laboratory data to support NEC diagnosis and personalized surgical decision-making. IMPACT:This study developed a dual Swin Transformer, which integrates abdominal X-rays and laboratory data to provide a robust multimodal framework for the diagnosis and surgical prediction of necrotizing enterocolitis. By implementing an external data domain adaptation strategy, the study contributes to overcoming the key challenge of clinical heterogeneity and temporal variability in NEC cohorts. Using Grad-CAM and SHAP visualization to identify specific predictive characteristics improves model transparency and clinician trust. These findings provide an explainable and adaptable AI tool to support evidence-based and personalized clinical decision-making in neonatal intensive care.

BACKGROUND:Digestive congenital anomalies (DCAs) are a major cause of neonatal death and disability, but comprehensive data on their global burden and regional disparities are limited. This study aimed to evaluate the global trends and burden of DCAs from 1990 to 2021. METHODS:Data from Global Burden of Disease Study 2021 were used to estimate annual prevalence, mortality, disability-adjusted life years (DALYs), age-standardized rates (ASPR, ASMR, ASDR) of DCAs. The average annual percent change (AAPC) was calculated and frontier analysis was conducted to evaluate the potential for burden reduction. Future trends were projected using statistical modeling. RESULTS:Between 1990 and 2021, the global burden of DCAs decreased significantly, with reductions in mortality and DALYs. ASMRs declined from 1.19 (0.66, 1.85) to 0.77 (0.58, 0.96) per 100,000 (AAPC = -1.40), and ASDRs from 108.30 (60.88, 166.69) to 70.44 (53.14, 87.60) (AAPC = -1.38). The burden was highest in infants aged 0-6 days and in low-SDI regions, which have greater improvement potential. By 2036, prevalent cases will increase, but deaths and DALYs are expected to continue declining. CONCLUSIONS:Despite progress in reducing the burden of DCAs, disparities persist, particularly in low-SDI regions, emphasizing the need for improved healthcare and neonatal surgery access. IMPACT:We analyzed global trends in the prevalence, mortality, and DALYs of digestive congenital anomalies (DCAs) from 1990 to 2021 using data from the Global Burden of Disease Study 2021. Our findings reveal that although the global burden of DCAs has declined, significant disparities persist, particularly in low- and middle-SDI regions. Frontier analysis emphasized substantial improvement potential in these regions. These results underscore the urgent need for enhanced screening programs, healthcare resources, and targeted interventions in low-SDI regions to further reduce the burden of DCAs and achieve equitable healthcare outcomes.

BackgroundGroup 2 innate lymphoid cells (ILC2) are the main group of tissue-resident ILCs in the lungs, which protect airway barrier integrity following infection. Macrophages are integral to the regulation of immune homeostasis in sepsis. However, the relationship between ILC2 and macrophages in the context of sepsis induced acute lung injury remains uncertain.MethodsThe sepsis was conducted by cecal ligation and puncture (CLP) model in Wild Type (WT) mice and ILC2 depleted mice. Septic mice were injected intratracheally IL-9, and the frequency and markers expression of ILC2 and macrophage were measured by Flow cytometry and CyTOF. The lung injury was conducted with pathological analysis. In vitro studies, MH-S cells were exposed to LPS with/without interleukin-9 (IL-9), and mTOR level and MH-S cells death were measured with western bloting or Flow cytometry.ResultsSepsis induced the accumulation of ILCs and pulmonary macrophages in lungs. Furtherly, we revealed that ILC2 and CD45+F4/80+CD11c+ macrophages expanded during sepsis induced acute lung injury. Meanwhile, ILC2 depletion significantly enhanced macrophages expansion. In vivo and in vitro studies determined that pulmonary macrophage death followed by sepsis were protedced by IL-9, which was main secreted by ILC2 in lung. Furthermore, IL-9 significantly declined the expression of mTOR, and the presence of ILC2 or IL-9 reduced the expression of M1 markers (CD86 or MHC II).ConclusionsIL-9 secreted by ILC2 has a protective role in sepsis induced lung injury by reducing macrophage apoptosis and M1 polarization via mTOR.

The upper airway morphology in children varies with age and body position. This study aimed to analyze the impact of lateral positioning on the upper airway of sedated children under five. This retrospective study included pediatric patients who underwent MRI in both the supine and lateral positions at Children’s Hospital, Zhejiang University School of Medicine. Upper airway morphology was reconstructed using 3D Slicer software. Python was employed to estimate cross-sectional areas via pixel analysis. The narrowest cross-sectional area, minimal transverse and anteroposterior diameters, airway length, and airway volume were measured and stratified by age for subgroup analysis. In sedated children under 5 years old and when compared to the supine position, lateral positioning increased minimal transverse diameter by 18.70

Background: Mesenchymal stem cells (MSCs) inhibit macrophage inflammatory response and alleviate intestinal inflammation. However, the role of MSCs in Hirschsprung-associated enterocolitis (HAEC) remains uncertain. This study aims to investigate the effects of MSCs on HAEC and the mechanisms related to macrophages and MSCs. Methods: Immunofluorescence was used to measure CD68 and p-AKT in colonic tissues of HSCR patients with HAEC. Ednrb-/- mice was used as HSCR model. The proportion of colonic tissue macrophages in WT and Ednrb-/- mice was assessed by flow cytometry. The colonic tissues injury was evaluated with HE staining and the survival curves of mice were recorded. In vitro, macrophage-induced enterocyte death was induced by lipopolysaccharide (LPS). MSCs, MSC derived exosomes, miR-223, or MK2206 were added to macrophages, and the levels of miR-223 in macrophages after exosome treatment were measured by RT-qPCR. Flow cytometry was used to assess enterocyte death, western blot was performed to measure p-AKT expression in macrophages, and enzyme-linked immunosorbent assay (ELISA) was used to detect IL-1β concentration in macrophage supernatants and serum of Edrnb -/- mice. Results: Increased expression of CD68 and p-AKT was observed in the colonic tissues of HAEC patients. Colonic instillation of MSCs derived exosomes significantly reduce the inflammatory score of colonic tissues and prolong the survival time of HAEC mice. In vitro, LPS-stimulated macrophages induce the phosphorylation of AKT and enterocyte death. Stimulation of macrophages with MSC-derived exosomes increased the content of miR-223. MSC-derived exosomes, miR-223 and MK2206 significantly reduce macrophage-induced enterocyte death, attenuated AKT phosphorylation in macrophages, and decreased IL-1β concentration in macrophage supernatants. Conclusion: Macrophages accumulate in colonic tissues during HAEC and inflammatory macrophages drive enterocyte death. MSCs derived exosomes reduce enterocyte death by suppressing AKT phosphorylation and IL-1β secretion via miR-223, and subsequently mitigate HAEC in mice. These findings suggest that MSC-derived exosomes, particularly those enriched in miR-223, may serve as a promising therapeutic strategy for the prevention or treatment of HAEC.

To analyze the clinical characteristics and available treatment strategies for reoperation of neonatal high jejunal atresia, and recommend preventive measures to reduce the reoperation rate of high jejunal atresia. The clinical data of 16 children with high jejunal atresia who underwent reoperation in the Neonatal Surgery Department at Children’s Hospital of Zhejiang University School of Medicine from January 2018 to January 2023 were retrospectively analyzed. Among the 16 unplanned reoperations, 7 (43.6

The respiratory tract related diseases are the main causes of death in children. Child pulmonary severe infection is the most important inducement for Respiratory tract related diseases. Therefore, it is necessary to understand the composition of the microbial community and the species correlation in children with severe pulmonary infection. In this study, the bronchoalveolar lavage fluid (BALF), blood and cerebrospinal fluid (CSF) samples of 782 children with severe infection in the early stage were systematically analyzed by amplicon-sequencing technology to reveal the distribution of microbial community and its clinical correlation. Results reveal significantly higher bacterial abundance in BALF than in blood/cerebrospinal fluid, with community structures shaped by tissue microenvironments, geographical disparities, age, and clinical symptoms. Neonatal BALF harbors simplified microbiota dominated by probiotics, while diversity increases with age, showing marked differences between 1-3 and 6-12-year-olds. Sepsis samples exhibit reduced microbial diversity, with Staphylococcus and Moraxella enrichments in the CST-4 cluster. Respiratory symptom progression correlates with microbiota succession from oral colonizers to respiratory pathogens. Third-generation sequencing-derived co-occurrence networks illustrate synergistic interactions among opportunistic pathogens, providing a basis for ecological targeted therapy. This study provides a key basis for breaking through the limitations of traditional diagnosis, establishing a precise diagnosis and treatment system based on third-generation sequencing, and developing ecological targeted treatment strategies.

Objective: Open Ladd's procedure remains the gold standard for intestinal malrotation correction, while laparoscopic techniques have advanced significantly. However, reports on robot-assisted surgery for this condition are lacking. This study evaluates our experience with robot-assisted surgery for malrotation, focusing on its feasibility, safety, and effectiveness. Methods: We performed a retrospective analysis of patients who underwent robot-assisted surgery for intestinal malrotation at our hospital from January 2019 to December 2022. Data on surgical procedures, perioperative management, and follow-up outcomes were collected. Results: A total of 45 patients (mean age: 11.7 days) underwent robot-assisted surgery without conversion to open procedures. All patients were symptomatic, with surgeries completed in an average of 57 min (range: 42-78). The intra-abdominal pressure (IAP) was 5.6 mmHg (range: 5-7), and the endtidal carbon dioxide (ETCO2) was 38.4 mmHg (range: 35-45). Mechanical ventilation lasted an average of 5.2 h (range: 4-9). Initial oral feeding commenced at 3.1 days (range: 2-6) post-surgery, with sufficient feeding achieved by 7.2 days (range: 5-18). The average hospital stay was 12.1 days (range: 9 -27). One patient experienced prolonged hospitalization (27 days) due to abnormal intestinal motility. At an average follow-up of 11 months (range: 6-18), one (2.2 %) patient required redo surgery for obstruction, and three (6.7 %) had incision infections. The remaining 41 patients recovered without complications such as vomiting or abdominal distension. Conclusion: Robot-assisted surgery for intestinal malrotation is a reliable and feasible approach, yielding favorable surgical outcomes. (c) 2025 Asian Surgical Association and Taiwan Society of Coloproctology. Publishing services by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/ by-nc-nd/4.0/).

Macrophages are vital in maintaining tissue homeostasis in the lungs by modulating and regulating immune responses. Based on different origins and anatomical locations, macrophages in the lungs are categorized into alveolar macrophages, interstitial macrophages, perivascular macrophages, and inflammatory macrophages. Alveolar macrophages are located in the alveolar spaces and are primarily responsible for maintaining alveolar surfactant homeostasis, defending against pathogens and regulating immune responses. Interstitial macrophages can maintain homeostasis, regulate immunity and anti-inflammation in the lung tissue. Perivascular macrophages play a crucial role in inhibiting lung inflammation, improving pulmonary fibrosis, and regulating lung tumor progression due to antigen-presenting and immunomodulatory effects. Inflammatory macrophages, which are differentiated from monocytes during inflammation, regulate the inflammatory process. This article reviews the origins of various subpopulations of macro-phages in the lung tissue and their physiological and pathological functions as well as discusses the underlying mechanisms and potential therapeutic targets.

Abstract The timely detection and management of hemorrhagic shock hold paramount importance in clinical practice. This study was designed to establish a nomogram that may facilitate early identification of hemorrhagic shock in pediatric patients with multiple-trauma. A retrospective study was conducted utilizing a cohort comprising 325 pediatric patients diagnosed with multiple-trauma, who received treatment at the Children's Hospital, Zhejiang University School of Medicine, Zhejiang, China. For external validation, an additional cohort of 144 patients from a children's hospital in Taizhou was included. The model's predictor selection was optimized through the application of the Least Absolute Shrinkage and Selection Operator (LASSO) regression. Subsequently, a prediction nomogram was constructed using multivariable logistic regression analysis. The performance and clinical utility of the developed model were comprehensively assessed utilizing various statistical metrics, including Harrell's Concordance Index (C-index), receiver operating characteristic (ROC) curve analysis, calibration curve analysis, and decision curve analysis (DCA). Multivariate logistic regression analysis identified systolic blood pressure (ΔSBP), platelet count, activated partial thromboplastin time (APTT), and injury severity score (ISS) as independent predictors for hemorrhagic shock. The nomogram constructed using these predictors demonstrated robust predictive capabilities, as evidenced by an impressive area under the curve (AUC) value of 0.963. The model's goodness-of-fit was assessed using the Hosmer–Lemeshow test (χ2 = 10.023, P = 0.209). Furthermore, decision curve analysis revealed significantly improved net benefits with the model. External validation further confirmed the reliability of the proposed predictive nomogram. This study successfully developed a nomogram for predicting the occurrence of hemorrhagic shock in pediatric patients with multiple trauma. This nomogram may serve as an accurate and effective tool for timely and efficient management of children with multiple trauma.

Abstract Background Postoperative delirium (POD) represents a prevalent and noteworthy complication in the context of pediatric surgical interventions. In recent times, a hypothesis has emerged positing that cerebral ischemia and regional cerebral oxygen desaturation might serve as potential catalysts in the pathogenesis of POD. The primary aim of this study was to methodically examine the potential relationship between POD and regional cerebral oxygen saturation (rSO2) and to assess the predictive and evaluative utility of rSO2 in the context of POD. Methods This prospective observational study was conducted at the Children’s Hospital, Zhejiang University School of Medicine, Zhejiang, China, spanning the period from November 2020 to March 2021. The research cohort comprised children undergoing surgical procedures within this clinical setting. To measure rSO2 dynamics, cerebral near-infrared spectroscopy (NIRS) was used to monitor rSO2 levels both before and after surgery. In addition, POD was assessed in the paediatric patients according to the Diagnostic and Statistical Manual of Mental Disorders Fifth Edition (DSM-5) criteria. The analysis of the association between the rSO2 index and the incidence of POD was carried out through the application of either the independent samples t-test or the nonparametric rank-sum test. To ascertain the threshold value of the adjusted rSO2 index for predictive and evaluative purposes regarding POD in the pediatric population, the Receiver Operating Characteristics (ROC) curve was employed. Results A total of 211 cases were included in this study, of which 61 (28.9%) developed POD. Participants suffering delirium had lower preoperative rSO2mean, lower preoperative rSO2min, and lower postoperative rSO2min, higher ∆rSO2mean, higher amount of ∆rSO2mean, lower ∆rSO2min (P < 0.05). Preoperative rSO2mean (AUC = 0.716, 95%CI 0.642–0.790), ∆rSO2mean (AUC = 0.694, 95%CI 0.614–0.774), amount of ∆rSO2mean (AUC = 0.649, 95%CI 0.564–0.734), preoperative rSO2min (AUC = 0.702, 96%CI 0.628–0.777), postoperative rSO2min (AUC = 0.717, 95%CI 0.647–0.787), and ∆rSO2min (AUC = 0.714, 95%CI 0.638–0.790) performed well in sensitivity and specificity, and the best threshold were 62.05%, 1.27%, 2.41%, 55.68%, 57.36%, 1.29%. Conclusions There is a close relationship between pediatric POD and rSO2. rSO2 could be used as an effective predictor of pediatric POD. It might be helpful to measure rSO2 with NIRS for early recognizing POD and making it possible for early intervention.

AIMS:Neonatal necrotizing enterocolitis (NEC) is a leading cause of intestine inflammatory disease, and macrophage is significantly activated during NEC development. Posttranslational modifications (PTMs) of proteins, particularly ubiquitination, play critical roles in immune response. This study aimed to investigate the effects of ubiquitin-modified proteins on macrophage activation and NEC, and discover novel NEC-related inflammatory proteins. MATERIALS AND METHODS:Proteomic and ubiquitin proteomic analyses of intestinal macrophages in NEC/healthy mouse pups were carried out. In vitro macrophage inflammation model and in vivo NEC mouse model, as well as clinical human samples were used for further verification the inhibitor of nuclear factor-κB kinase α (IKKα) ubiquitination on NEC development through Western blot, immunofluorescence, quantitative real-time polymerase chain reaction (qRT-PCR) and flow cytometry. KEY FINDINGS:We report here that IKKα was a new ubiquitin-modified protein during NEC through ubiquitin proteomics, and RING finger protein 31 (RNF31) acted as an E3 ligase to be involved in IKKα degradation. Inhibition of IKKα ubiquitination and degradation with siRNF31 or proteasome inhibitor decreased nuclear factor-κB (NF-κB) activation, thereby decreasing the expression of pro-inflammatory factors and M1 macrophage polarization, resulting in reliving the severity of NEC. SIGNIFICANCE:Our study suggests the activation of RNF31-IKKα-NF-κB axis triggering NEC development and suppressing RNF31-mediated IKKα degradation may be therapeutic strategies to be developed for NEC treatment.

BACKGROUND:Neonatal necrotizing enterocolitis (NEC) is one of the most prevalent and severe intestinal emergencies in newborns. The inflammatory activation of macrophages is associated with the intestinal injury of NEC. The neuroimmune regulation mediated by α7 nicotinic acetylcholine receptor (α7nAChR) plays an important role in regulating macrophage activation and inflammation progression, but in NEC remains unclear. This study aims to explore the effect of macrophage α7nAChR on NEC. METHODS:Mice NEC model were conducted with high-osmolarity formula feeding, hypoxia, and cold stimulation. The α7nAChR agonist PNU-282987 and mTOR inhibitor rapamycin were treated by intraperitoneal injections in mice. The expression and distribution of macrophages, α7nAChR, and phospho-mammalian target of rapamycin (p-mTOR) in the intestines of NEC patients and mice was assessed using immunohistochemistry, immunofluorescence, and flow cytometry. The expression of NLRP3, activated caspase-1 and IL-1β in mice intestines was detected by flow cytometry, western blot or ELISA. In vitro, the mouse RAW264.7 macrophage cell line was also cultured followed by various treatments. Expression of p-mTOR, NLRP3, activated caspase-1, and IL-1β in macrophages was determined. RESULTS:Macrophages accumulated in the intestines and the expression of α7nAChR in the mucosal and submucosal layers of the intestines was increased in both the NEC patients and mice. The p-mTOR and CD68 were increased and co-localized in intestines of NEC patients. In vitro, α7nAChR agonist PNU-282987 significantly reduced the increase of NLRP3, activated caspase-1, and IL-1β in macrophages. PNU-282987 also significantly reduced the increase of p-mTOR. The effect was blocked by AMPK inhibitor compound C. The expression of NLRP3, activated caspase-1, and IL-1β was inhibited after mTOR inhibitor rapamycin treatment. In NEC model mice, PNU-282987 reduced the expression of p-mTOR, NLRP3, activated caspase-1, and IL-1β in the intestine. Meanwhile, rapamycin significantly attenuated NLRP3 activation and the release of IL-1β. Moreover, the proportion of intestinal macrophages and intestinal injury decreased after PNU-282987 treatment. CONCLUSION:Macrophage α7nAChR activation mitigates NLRP3 inflammasome activation by modulating mTOR phosphorylation, and subsequently alleviates intestinal inflammation and injury in NEC.

Sepsis is a life-threatening organ malfunction induced by an imbalanced immunological reaction to infection in the host. Many studies have utilized traditional RNA sequencing (RNA-seq) data to identify important biological targets to predict sepsis prognosis. However, alterations in core cells and functional status cannot be effectively detected in sepsis patients. The goal of this study was to identify key cells through single-cell RNA-seq (scRNA-seq), and combine bulk RNA-seq data and multiple algorithm analysis to construct a stable prognostic model for sepsis. The scRNA-seq and bulk RNA-seq data from sepsis patients were collected from the Gene Expression Omnibus (GEO) database. The R package "Seurat" was used to process the scRNA-seq data. Cell communication was investigated using the R package "CellChat". The pseudo-time of the cells was calculated using the R package "monocle". The R package "limma" was used to identify differentially expressed genes (DEGs) between the sepsis group and the control group. Weighted gene correlation network analysis (WGCNA) was used to identify critical modules. Eight kinds of machine learning and 90 algorithm combinations were used to construct the prognostic model for sepsis. Quantitative real-time PCR (qRT-PCR) was performed to determine the expression of key genes in the cecal ligation and puncture (CLP)-induced sepsis mouse model. The immunological status and related properties of DEGs were then investigated in the high- and low-risk groups delineated by the model. By combining the scRNA-seq data from nine samples, 13 clusters and 9 cell types were identified. CellChat analysis revealed that the number and strength of interactions between platelets and a variety of cells increased. We identified key platelet genes from the scRNA-seq data and combined these genes and the results of differential analysis and WGCNA of the bulk RNA-seq data. After univariate Cox regression analysis, we calculated the Cindex of the model constructed by the combination of 90 algorithms, and we finally determined the "CoxBoost + Lasso" combination. Multivariate Cox regression was used to construct the final prognostic model. The qRT-PCR results revealed significant differences in five key prognostic genes between the CLP and sham groups. The data was classified into high- and low-risk groups based on the model score. The high-risk group had a poorer survival rate and less immune infiltration. We identified the importance of platelets in sepsis patients through scRNA-seq, and established prognostic models with key genes that were identified via scRNA-seq combined with bulk RNA-seq analysis. The results of this model were closely associated with patient survival rates and immunological status and this model is useful for the prognostic management of sepsis.

The purpose of this study was to retrospectively compare the short-term outcomes of robotic- (RAD) and laparoscopic-assisted duodenal diamond-shaped anastomosis (LAD) in neonates. Neonates who underwent RAD (n = 30) or LAD (n = 38) between January 2019 and December 2022 were analyzed retrospectively. Major patient data were collected, including preoperative, intraoperative, and postoperative information. All patients were neonates below the age of 30 days weighing 4 kg. Thirty (44.1