Glucocorticoid insensitivity affects a subset of asthma patients, yet its mechanism is unclear. Given the key role of airway epithelium in asthma, this study hypothesized that glucocorticoids may impair this barrier. We examined whether glucocorticoid-induced autophagy-associated apoptosis contributes to reduced treatment efficacy. Using an ovalbumin-induced asthma model in BALB/c mice and 16HBE human bronchial epithelial cells treated with dexamethasone and/or autophagy inhibitor EACC (C13H11N3O6S2), we assessed cell viability and apoptosis via CCK-8 and TUNEL assays, respectively. Apoptosis and autophagy markers were analyzed by quantitative polymerase chain reaction (PCR) and western blot. Dexamethasone alleviated airway inflammation but worsened epithelial integrity in asthmatic mice. It failed to downregulate pro-apoptotic factors while reducing anti-apoptotic factors both in vivo and in vitro. Dexamethasone decreased 16HBE cell viability and increased apoptosis. Mechanistically, dexamethasone upregulated autophagy markers and suppressed anti-autophagy factors in mouse lungs and cells, concurrently reducing the phosphorylation of negative regulators of autophagy. Importantly, the autophagy inhibitor EACC enhanced cell viability and attenuated dexamethasone-induced apoptotic signaling in 16HBE cells. These findings suggest glucocorticoids may compromise the airway epithelial barrier via autophagy-associated apoptosis in vitro and in vivo, indicating autophagy inhibition as a potential hypothesis for future therapeutic exploration for glucocorticoid-insensitive asthma.
Among preterm infants, bronchopulmonary dysplasia (BPD) is the most prevalent chronic pulmonary disorder. Its pathogenesis involves a complex interplay between prenatal inflammatory exposure and abnormal immune activation. Neutrophil extracellular traps (NETs) are mediators of tissue damage, but whether they bridge intrauterine inflammation and the subsequent development of BPD is not yet clear. We used a prenatal lipopolysaccharide (LPS)-exposed rat model to ask two questions: first, whether excessive NETs drive BPD pathogenesis, and second, whether human umbilical cord mesenchymal stem cell-derived microvesicles (hUCMSC-MVs) can alleviate lung injury by inhibiting this process. We established a BPD model in preterm rats via prenatal LPS exposure and assessed lung morphometry, neutrophil infiltration, and NETs markers. We then compared the therapeutic effects of hUCMSC-MVs with NET extrusion (NETosis) inhibitors in vivo and also examined direct neutrophil effects in vitro. Prenatal LPS exposure led to significantly elevated NET markers in both lung tissue and circulation, which correlated with severe alveolar simplification. Neutrophils in this model exhibited a "primed" phenotype with enhanced potential for NETs release. In vivo, hUCMSC-MVs suppressed NET formation, attenuated neutrophilic inflammation, and restored alveolar structure, matching the efficacy of conventional NETosis inhibitors. In vitro, they were internalized by neutrophils and abrogated LPS-induced NETosis. These findings point to excessive NETosis as a key mechanistic link between prenatal inflammation and postnatal lung impairment. hUCMSC-MVs directly inhibit this pathological process and alleviate BPD-like lung injury. This work advances the mechanistic understanding of BPD and supports hUCMSC-MVs as a promising cell-free therapeutic candidate.
Tubular epithelial dedifferentiation and loss of polarity are early pathological events in obstructive nephropathy and critically contribute to progressive renal fibrosis. Human umbilical cord mesenchymal stem cell-derived exosomes (HucMSC-Exos) have shown therapeutic potential for kidney injury, yet the mechanisms by which they preserve epithelial identity remain unclear. Mafb, a transcription factor essential for renal epithelial differentiation, has been implicated in epithelial homeostasis, but its role in adult obstructive injury and its regulation by exosomes are unknown. Here, a unilateral ureteral obstruction (UUO) mouse model was used to evaluate early tubular injury and the effects of HucMSC-Exos. UUO induced marked epithelial damage characterized by decreased E-cadherin, increased vimentin and α-smooth muscle actin (α-SMA) expression, and disruption of epithelial polarity, whereas HucMSC-Exo administration significantly ameliorated these alterations. Mafb expression was profoundly downregulated in UUO kidneys but was effectively restored following exosome treatment. In vitro, Mafb knockdown in Human renal proximal tubular epithelial (HK-2) cells markedly aggravated TGF-β1-induced dedifferentiation, confirming its protective role in epithelial integrity. Analysis of Mafb -/- embryonic kidneys demonstrated severe developmental defects and loss of polarity markers, highlighting the essential function of Mafb in renal epithelium. Importantly, HucMSC-Exos rescued epithelial marker expression even in Mafb-silenced cells, suggesting both Mafb-dependent and independent mechanisms. These findings identify Mafb as a key regulator of tubular epithelial homeostasis and reveal that HucMSC-Exos attenuate early renal injury partly through Mafb restoration.
The optimal dose for ultrasound-guided axillary brachial plexus block (ABPB) in children remains poorly defined. This study aimed to find the median effective volume (EV50) of ropivacaine (0.2%) for ABPB in children aged 6-10 years. A prospective, double-blind, dose-finding trial using the Dixon up-and-down method was conducted. Pediatric patients (aged 6-10 years, ASA I-II) undergoing unilateral elbow or below-elbow surgery were enrolled. Ultrasound-guided ABPB was performed using a dual-injection technique with supplemental musculocutaneous nerve blockade. The initial volume of ropivacaine (0.2%) was 0.5 mL/kg, adjusted by 0.05 mL/kg based on the previous patient's response. The study continued until seven inflection points were observed from failure to success. EV50 and EV95 (95% effective volume) were calculated using isotonic regression and bootstrapping, with 95% confidence intervals (CIs). Patient demographics, postoperative pain scores, and adverse events were monitored. Twenty-nine patients were enrolled. The EV50 was 0.350 mL/kg (95% CI: 0.197-0.362 mL/kg), and the EV95 was 0.395 mL/kg (95% CI: 0.385-0.396 mL/kg). No adverse events were observed during the perioperative period. This study provides evidence-based dosing parameters for 0.2% ropivacaine in pediatric ABPB, aiding clinicians in optimizing efficacy and safety. The findings support age-specific precision in pediatric regional anesthesia.
Hypospadias is among the most frequent congenital malformations of the male genitalia. The expression of V-maf musculoaponeurotic fibrosarcoma oncogene homolog B (MAFB), a transcription factor within the androgen signaling pathway, is closely correlated with the etiology of this condition. From an epigenetic perspective, this study investigated the role of microRNA-130a-3p (miR-130a-3p) in regulating MAFB during hypospadias pathogenesis. Clinical foreskin samples from hypospadias children and the dioctyl phthalate (DEHP)-induced mouse genital tubercle tissues were analyzed. The expression levels of miR-130a-3p and MAFB were detected by qRT-PCR and western blot. In vitro, miR-130a-3p was modulated in HS68 cells; CCK-8, scratch assay, western blot, flow cytometry, and dual-luciferase assay were used to assess its effects and targeting of MAFB. Our study demonstrated that miR-130a-3p targeted and suppressed MAFB expression. This suppression inhibited cellular proliferation and migration and reduced Vimentin mRNA expression. Cell cycle distribution was disrupted, marked by Gap 1 phase increase and Synthesis phase reduction, and the cell apoptosis rate increased. Cyclin-dependent kinase 2 (CDK2), Cyclin E1, and Proliferating Cell Nuclear Antigen (PCNA) were downregulated. miR-130a-3p targets and inhibits MAFB expression, disrupting the normal processes of cell proliferation, migration, cell apoptosis, and cell cycle progression, ultimately leading to the development of hypospadias.
The aim of this study was to investigate the potential reproductive toxicity of bisphenol S (BPS) and the related molecular mechanisms through a network toxicology approach. By utilizing various databases, including the Comparative Toxicogenomics Database (CTD), GeneCards, Online Mendelian Inheritance in Man (OMIM), the Pharmacogenomics Knowledgebase (PharmGKB), and the Therapeutics Target Database (TTD), and limiting the species to Homo sapiens, we confirmed 45 potential targets related to both BPS exposure and reproductive injury. Additional analysis via STRING and Cytoscape pinpointed 4 key targets: AKT serine/threonine kinase 1 (AKT1), interleukin-6 (IL-6), interleukin-1-beta (IL-1β), and tumor necrosis factor α (TNFα). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses indicated that the principal targets of BPS-mediated reproductive damage are mainly involved in oxidative stress signaling and cell secretion pathways. Because PI3K/AKT regulates the secretion of IL-6, IL-1β, and TNFα, we focus on AKT1 in the molecular docking experiments. Unexpectedly, we found a strong interaction between BPS and AKT1. Furthermore, the results suggested that women may be more susceptible to BPS-mediated reproductive toxicity than men. This study provides a theoretical framework for understanding the molecular mechanisms through which BPS causes reproductive toxicity and lays the foundation for preventing and treating reproductive disorders related to BPS exposure. In addition, our network toxicology-based method can accelerate the discovery of pathways involved in the toxic effects of environmental chemicals that are currently unknown.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) may impact immune-related micro ribonucleic acids (miRNAs). The role of salivary miRNAs as predictors of severe infection in children is unknown. This study sought to examine the relationship between SARS-CoV-2 severity and salivary miRNA levels in children. A convenience sample of 400 children with SARS-CoV-2 infection was prospectively evaluated between March 2021 and February 2022 at two U.S. children's hospitals. Saliva swabs were obtained. Levels of 3 miRNAs previously implicated in SARS-CoV-2 pathophysiology were measured with quantitative polymerase chain reaction (miR-1273c, miR-296-5p, and miR-4495). The primary outcome measure was the occurrence of severe illness, including respiratory failure, shock, or death from a related cause within 1 month of evaluation. Disease resolution was determined through chart review and caretaker phone calls 4 weeks after the initial visit. Saliva samples from 395 (severe = 105) participants (mean age: 7.4 ± 5.9 years, 51.7% female, 54.7% African American) were analyzed. Children with severe outcomes had lower levels of miR-296-5p (fold difference: 0.361, d = 0.111, p = 0.046). Multivariable adjusted logistic regression analysis demonstrated an inverse association between severe outcomes and levels of miR-296-5p (adjusted odds ratio = 0.89; 95% confidence interval [0.81, 0.98], p = 0.021) while controlling for age, sex, race, weight, insurance, diabetes, asthma, fever, and symptom duration. The model had an area under curve of 0.744 (sensitivity = 0.71, specificity = 0.66). Measurement of saliva miRNA in conjunction with demographic and clinical characteristics may aid in the prediction of severe illness. Larger, longitudinal studies to assess the utility of salivary miRNAs in SARS-CoV-2 and other viral infections are needed.
Childhood asthma remains a global public health challenge with suboptimal control, despite the availability of inhaled corticosteroids (ICS) as first-line therapy. Glucocorticoids suppress inflammation and protect the airway epithelial barrier (AEB) via genomic and nongenomic pathways. However, a significant subset of patients, particularly those with severe asthma, exhibits glucocorticoid resistance. Emerging evidence reveals a dual role of glucocorticoids on the AEB: beyond protection, they may inadvertently compromise barrier integrity by inducing epithelial apoptosis and dysfunction. This review synthesizes current knowledge on glucocorticoid mechanisms in asthma, focusing on the AEB as a critical interface between therapeutic efficacy and treatment failure. We show that AEB impairment serves as one of the key mechanisms underpinning glucocorticoid resistance and the progression to severe asthma, with heightened relevance in children due to the unique vulnerability of their developing airways. Furthermore, we examine how infections, nutritional factors (e.g., vitamins A and D), and immune maturation intersect with AEB integrity and glucocorticoid responsiveness. By reframing the AEB as both a target and a determinant of glucocorticoid efficacy, this review highlights the urgent need for barrier-focused strategies to overcome resistance and improve outcomes in childhood asthma.
The specific mechanisms and screening methods for congenital heart disease (CHD) remain elusive. Evidence indicates that aberrant maternal metabolomic profiles are associated with CHD, but it is uncertain whether such an association exists in umbilical cord blood. This study aimed to measure the metabolome shifts in monozygotic (MZ) twins discordant for CHD and, if present, identify the altered metabolites and metabolic pathways. Umbilical cord blood from three pairs of MZ twins discordant for CHD, identified through the prospective, population-based longitudinal twin study (LoTiS), was subjected to ultrahigh-performance liquid chromatography coupled with mass spectrometry (UHPLC-MS/MS) based metabolomic analysis. Orthogonal partial least square-discriminate analysis (OPLS-DA) and random forest (RF) analysis were used to determine differences in metabolic profiles and individual metabolites. In univariate analysis, two specific metabolites were identified in the umbilical cord blood of CHD cases compared to their MZ twins without complications. After the paired-sample t-test, four differentially expressed metabolites (DEMs) were identified, three of which were closely related to fatty acids and their metabolism. Enrichment pathway analysis revealed dysregulation of various metabolic pathways, including glucose metabolism, lipid metabolism, and amino acid metabolism pathways, in MZ twins with CHD compared to their healthy counterparts. The results demonstrated that the metabolomic signature of umbilical cord blood in CHD differs from that of their healthy MZ co-twins, and the revelation of associated metabolites and pathways provide preliminary clues for generating hypotheses about the metabolic correlates of CHD predisposition. Findings are limited by small sample size and require validation in larger, independent cohorts.
ABSTRACT Cleft lip (CL) and cleft palate (CP), collectively referred to as orofacial clefts (OFCs), are among the most common birth defects and can have significant effects on speech, nutrition, and physical and psychosocial development. Manifestation, classification, and treatment plans of OFCs are diverse and not standardized. Recent technological advancements and genetic testing have enabled researchers to propose relationships between certain genes and the development of OFCs. Potential genetic precursors for OFCs have been found to be numerous and diverse. Treatment strategies are also numerous and vary greatly between individuals. In this review article, we summarize the most commonly discussed genetic factors in the literature, variation in classification, and advances in management.
The global prevalence of heart failure in children continues to increase, accompanied by a growing disease burden. To advance early prevention and intervention by shifting the focus upstream in the disease process, the Cardiovascular Group of the Pediatrics Society of the Chinese Medical Association has introduced the concept of pediatric cardiac function staging applicable to all children. This framework categorizes pediatric cardiac function into three distinct stages: the stage of cardiac function sufficiency (Stage A), the stage of cardiac dysfunction (Stage B), and the stage of cardiac function failure (Stage C). Based on this staging system, the diagnostic criteria and management strategies for each stage are delineated, with the aim of establishing a comprehensive management pathway rooted in cardiac function staging. This approach seeks to further reduce the incidence and burden of pediatric heart failure, providing a theoretical foundation and practical guidance for achieving the goal of making pediatric heart failure preventable, manageable, and treatable.
X-linked hypophosphatemia (XLH) is a rare hereditary disorder characterized by PHEX gene mutations, elevated FGF23 levels, and impaired bone mineralization. Burosumab, a monoclonal antibody targeting FGF23, has demonstrated clinical efficacy; however, the immunological dynamics during treatment remain unexplored. This study employed longitudinal single-cell RNA sequencing (scRNA-seq) to characterize peripheral blood immune cell alterations across multiple treatment stages in pediatric XLH. We performed scRNA-seq on peripheral blood mononuclear cells from pediatric patients with XLH at five time points spanning pretreatment and burosumab therapy phases, along with healthy pediatric controls. A total of 93,112 cells were analyzed using comprehensive bioinformatic pipelines, including unsupervised clustering, pseudotime trajectory analysis, temporal gene expression profiling, and cell-cell communication inference. Eleven major immune cell populations were identified, with notable dynamic alterations in T cells and natural killer (NK) cell subtypes across treatment stages. The cellular proportion of T helper 2 (Th2) cells and regulatory T (Treg) cells were elevated before treatment and normalized during therapy, whereas T helper 17 (Th17) cells exhibited reciprocal patterns. Genes upregulated in Treg cells during early treatment were enriched in osteoclast differentiation pathway. Natural killer subtype 2 cells showed enrichment in osteoclast differentiation and interleukin-12 response pathways. Cell-cell communication analysis identified dynamic interactions among Th2 cells, Th17 cells, Treg cells, and NK cell subtypes mediated by KLRB1-CLEC2D and SELL-SELPLG ligand-receptor pairs. This longitudinal transcriptomic study provides the first comprehensive characterization of peripheral immune dynamics during burosumab therapy in XLH, offering new insights into the immunological mechanisms underlying treatment response.
Renal fibrosis is a common pathological feature and key driver of progression to end-stage renal disease in various chronic kidney diseases, with effective treatments remaining scarce. Exosomes derived from mesenchymal stem cells (MSC-Exos) have demonstrated tremendous potential in tissue and organ repair and antifibrosis treatment. This study investigated the therapeutic effects and mechanisms of MSC-Exo derived from human umbilical cord (HucMSC-Exo) on renal fibrosis. HucMSC-Exo was applied to intervene in a mouse model of renal fibrosis induced by unilateral ureteral obstruction (UUO) or cocultured with TGF-β-stimulated rat renal fibroblasts (NRK-49F). Results showed that HucMSC-Exo conspicuously alleviated pathological damage, inflammatory response, fibroblast proliferation/activation, and extracellular matrix deposition in UUO kidneys. Single-cell sequencing revealed a prominent upregulation of Inhba gene in UUO kidneys, whereas HucMSC-Exo treatment effectively inhibited its expression. Further mechanistic studies showed that knocking down Inhba mimicked the antifibrotic effects of HucMSC-Exo, whereas exogenously adding INHBA weakened its protective effects. Transcriptome sequencing results revealed that the downstream effects of Inhba involve regulating the PI3K/AKT signaling pathway, and HucMSC-Exo treatment markedly inhibited the activation of this pathway. Collectively, HucMSC‑Exo exerts antifibrotic effects by regulating the INHBA/PI3K/AKT signaling axis to inhibit renal fibroblast activation.
ABSTRACT Mild malnutrition in infants and young children is an early stage of nutritional deficiency characterized by a slight reduction in nutritional reserves and mild impairment of physiological functions but no obvious clinical symptoms or severe growth retardation. Parents often overlook this condition because typical signs are absent. This cross‐sectional study aimed to investigate the current status of mild malnutrition in infants and young children aged 6–24 months attending a pediatric health clinic, analyze its influencing factors, and provide a basis for nutritional health guidance. The study included 879 infants and young children aged 6–24 months who underwent physical examinations at the Outpatient Clinic of Pediatric Health Care, The Third Affiliated Hospital of Zhengzhou University, China, from December 2024 to August 2025. The survey results showed a detection rate of mild malnutrition of 13.88% (122/879). After including statistically significant factors from univariable analysis in a multivariable logistic regression, the following risk factors were identified: food allergies (odds ratio (OR) = 3.117, 95% confidence interval (CI): 1.902–5.108), low maternal education level (OR = 1.969, 95% CI: 1.166–3.325), low birth weight (OR = 1.759, 95% CI: 1.041–2.973), maternal prepregnancy BMI < 18.5 (OR = 2.197, 95% CI: 1.252–3.855), frequent nighttime awakenings (OR = 2.427, 95% CI: 1.481–3.975), and insufficient complementary feeding frequency (OR = 1.550, 95% CI: 1.030–2.332). The detection rate of mild malnutrition in infants and young children aged 6–24 months at pediatric clinics is relatively high, and clinical work should give sufficient attention to this issue. Proactive guidance should be provided to these children to promote healthy growth.
ABSTRACT The minimum alveolar concentration of sevoflurane varies with age in children, and the median effective concentration (EC50) of sevoflurane for I‐gel insertion in children of different ages has not been reported. The aim of this study was to determine the EC50 of end‐tidal sevoflurane maintained for 2.5 min for I‐gel insertion in children aged 1–10 years and also to estimate the 95% effective concentration (EC95). This study aimed to recruit children who were scheduled to undergo laparoscopic high ligation of the inguinal hernia sac or laparoscopic high ligature of the sheath process. Children were stratified into three age groups. We employed Dixon's up‐and‐down method in this study. The target end‐tidal sevoflurane concentration was then maintained at 2% for 2.5 min for the first child. The concentration for the subsequent patient was determined based on the response of the previous patient, with adjustments of 0.2%. This study was terminated when seven crossover points were reached. In the 1–3‐year‐old group including 3‐year‐old, EC50 was 1.75% (95% confidence interval [CI], 1.72%–2.03%), and EC95 was 2.17% (95% CI, 1.96%–2.18%). In the 3–6‐year‐old group including 6‐year‐old, EC50 was 1.60% (95% CI, 1.35%–1.83%), and EC95 was 1.96% (95% CI, 1.77%–1.98%). In the 6–10‐year‐old group, EC50 was 0.96% (95% CI, 0.93%–2.20%) and EC95 was 2.36% (95% CI, 2.15%–2.38%). Our study determined the EC50 of end‐tidal sevoflurane required for I‐gel insertion in unpremedicated children aged 1–10 years and the results demonstrate to be both safe and effective for pediatric patients.
The pathogenesis of heart failure involves a highly intricate process regulated by diverse epigenetic factors, transcription factors, noncoding RNAs, and cyclins. Notably, the reexpression of embryonic cardiac transcription factors, including GATA, MEF2, and Nkx2.5, is considered to exert critical influence in the initiation and advancement of heart failure. Nevertheless, the precise mechanisms through which epigenetic modifications drive this reprogramming of gene expression remain poorly defined. This investigation aims to clarify the role of histone acetylation in regulating the reexpression of embryonic cardiac transcription factors during heart failure. Our research indicates that during heart failure of mice, there are distinct histone acetylation modifications associated with the reexpression of these factors. Notably, GATA4 and MEF2C show significant increases, whereas Nkx2.5 shows a decrease compared to normal groups during heart failure progression. These findings imply that embryonic GATA4 and MEF2C may promote the development of heart failure, whereas Nkx2.5 does not appear to participate in disease progression. Furthermore, treatment with curcumin, a known inhibitor of histone acetylation, reduces acetylation levels at H3K4, H3K9, and H3K27 within the promoter regions of GATA4 and MEF2 C in a murine model of heart failure, leading to downregulation of these genes and subsequent enhancement of cardiac performance. In summary, our study demonstrates that p300 exerts site-specific regulatory effects on various transcription factors via histone modifications, and low acetylation status at specific sites can inhibit reactivation of GATA4 and MEF2C during the myocardial dysfunction period thereby improving cardiac performance of mice.
