ObjectiveThis study aims to investigate the molecular mechanism by which Guizhi Fulingwan （GFW） inhibits ferroptosis in endometriosis （EMT） through the regulation of the hypoxia inducible factor-1α/heme oxygenase 1 （HIF-1α/HO-1） signaling pathway. MethodsMachine learning was employed to identify ferroptosis-related biomarkers associated with EMT. Network pharmacology was utilized to identify the active components of GFW and its potential therapeutic targets against EMT， including core targets. Functional enrichment analysis was conducted to explore the biological processes， molecular functions， cellular components， and signaling pathways associated with the potential targets. An EMT rat model was established via autologous transplantation. Thirty female Sprague-Dawley （SD） rats were randomly divided into five groups： sham-operated， model， positive control （dienogest at 0.2 mg·kg^-1）， low-dose GFW （2.5 g·kg^-1）， and high-dose GFW （5 g·kg^-1）. After modeling， the rats received their respective treatment by oral gavage for 28 consecutive days， while the sham and model groups received equal volumes of distilled water. Serum and ectopic endometrial tissues were collected. Hematoxylin and eosin （HE） staining was employed to evaluate morphological alterations in ectopic lesions. Quantitative real-time polymerase chain reaction （Real-time PCR） and Western blot were conducted to assess mRNA and protein expression of HIF-1α， HO-1， glutathione peroxidase 4 （GPX4）， spermidine/spermine N1-acetyltransferase （SAT1）， and prostaglandin-endoperoxide synthase 2 （PTGS2）. Tissue levels of malondialdehyde （MDA）， glutathione （GSH）， and ferrous iron （Fe²⁺） were quantified using commercial assay kits. Serum levels of interleukin-6 （IL-6） and transforming growth factor-β₁ （TGF-β₁） were measured via enzyme-linked immunosorbent assay （ELISA）. ResultsFive ferroptosis-related biomarkers in EMT were identified： ALOX12， CHAC1， SAT1， AST1， and HO-1. Network pharmacology analysis revealed 42 active components of GFW and 192 potential therapeutic target genes related to EMT treatment， with FOS， JUN， HO-1 identified as core targets. Functional enrichment analysis indicated that the potential targets were primarily involved in oxidative stress response and reactive oxygen species metabolism and were enriched in the HIF-1 signaling pathway. Compared to the sham-operated group， the model group exhibited significant increases in both mRNA and protein expression of HIF-1α， HO-1， and PTGS2， as well as elevated tissue levels of Fe²⁺ and MDA. Conversely， GSH levels and the expression of GPX4 and SAT1 were markedly reduced， and serum levels of IL-6 and TGF-β₁ levels were significantly higher （P<0.01）. Compared with the model group， all GFW-treated groups showed significant downregulation of HIF-1α and HO-1， reduced Fe²⁺ levels， and downregulated expression of MDA， PTGS2， IL-6， and TGF-β₁. Meanwhile， GSH， GPX4， and SAT1 expression levels were significantly increased （P<0.05， P<0.01）， effectively ameliorating iron overload and oxidative stress， thereby demonstrating therapeutic efficacy in EMT， with the high-dose GFW demonstrating the most pronounced therapeutic effects. ConclusionGFW exerts therapeutic effects on endometriosis by regulating the HIF-1α/HO-1 signaling pathway to rectify iron metabolism disorders and attenuate free iron-induced oxidative damage. It upregulates the antioxidative defense system to inhibit lipid peroxidation cascades and modulates inflammatory cytokine networks. These effects collectively disrupt the pathological interaction between ferroptosis and chronic inflammation， providing a novel theoretical foundation for the clinical application of GFW in EMT treatment.

Objective:To investigate the method and clinical efficacy of simultaneous correction of sunken upper eyelid during double eyelid plasty.Methods:A retrospective analysis was conducted on patients with single eyelids and sunken upper eyelid treated at the Department of Plastic Surgery, the First People’s Hospital of Zhengzhou, from October 2022 to February 2024. All patients underwent incisional double eyelid plasty with individualized correction based on depression severity. For mild depression, a combination of pretarsal orbicularis oculi muscle flap folding and orbital septum fat flap transposition was performed for correction. For moderate depression, additional autologous stromal vascular fraction (SVF)-gel grafting would be performed if residual depression persisted after the aforementioned approach. For severe depression, if the orbital fat was sufficient, the orbicularis oculi muscle flap folding, orbital fat flap repositioning and autologous SVF-gel transplantation were used. If there was insufficient orbital fat, the correction involved the use of a orbicularis oculi muscle flap and SVF-gel transplantation. Postoperative follow-up was conducted regularly, and related complications were statistically analyzed. Six months after the surgery, two blinded plastic surgeons evaluated outcomes via visual analog scale (VAS), including five aspects: symmetry of palpebral folds(including width and curvature of the double eyelid fold), fold contour, depression improvement, scar quality, overall eyelid aesthetics. Each aspect was rated on a scale from 1 to 10, with higher scores indicating better outcomes. Patient satisfaction was scored separately (1-5 per item), with higher scores indicating greater satisfaction.Results:A total of 64 patients were enrolled, including 4 males and 60 females; the age ranged from 19 to 66 years[ (32.0±9.7) years]. The degree of sunken upper eyelid was mild in 33 cases, moderate in 19 cases and severe in 12 cases. All patients underwent double eyelid plasty and orbicularis oculi flap folding. Based on this, 45 cases (33 mild and 12 moderate cases) underwent orbital septum fat flap transposition, 11 severe cases underwent SVF-gel grafting, and 8 cases (7 moderate and 1 severe cases) required combined approaches to correct depression. During the procedure, 19 cases (38 sides) received (1.8±0.8) ml of SVF-gel injection. Among these cases, one case with severe sunken upper eyelid who did not receive orbital fat transfer was injected with high density fat (left side: 0.6 ml, right side: 0.8 ml) due to insufficient SVF-gel volume. Two cases with severe sunken upper eyelid who only had autologous SVF-gel transplantation did not fully correct the depression within 3 months post-surgery received a second SVF-gel injection[(1.0±0.1) ml]. Follow-up was conducted for 6 to 12 months after surgery. All patients experienced varying degrees of eyelid edema, which typically lasted 2 to 3 months. Ecchymosis (34 sides), conjunctival congestion (3 sides), and temporary ptosis (5 sides) all resolved within 2 weeks. Pigmentation (14 sides) and scar hyperplasia (3 sides) disappeared or stabilized within 6 months. No patients developed complications such as incision infection, hematoma, fat liquefaction, local skin unevenness, or induration. The surgeons’ VAS scores of the above five indicators were all > 8 points, and the satisfaction scores of patients for the five indicators were all > 4 points. In both scoring, the improvement of sunken upper eyelid scored the highest, which were (9.2 ± 0.9) points and (4.8 ± 0.6) points respectively.Conclusion:For patients with single eyelids and varying degrees of sunken upper eyelid, performing double eyelid plasty and orbicularis oculi muscle flap folding, individualized correction is achieved through autologous fat redistribution techniques, based on the severity of the sunken upper eyelid and the amount of orbital fat. This single procedure can restore upper eyelid volume and rejuvenate the appearance, resulting in a natural and aesthetically pleasing double eyelid with minimal complications and high patient satisfaction.

Objective To investigate the mechanism of Wenjing Decoction in treating endometriosis(EMT)using bioinformatics methods and in vitro experiments.Methods The active components and corresponding targets of Wenjing Decoction were obtained from the TCMSP database,while EMT-related targets were identified using the GEO database.Functional enrichment analysis was conducted on the targets to predict core targets for treating EMT with Wenjing Decoction.Molecular docking was performed on core targets-drug ligands,and in vitro experiments validated the findings.Results Through screening the TCMSP database,117 active components of Wenjing Decoction were identified,corresponding to 248 targets;5 312 EMT-related differential genes were gathered from GEO database,identifying 97 potential targets of Wenjing Decoction for treating EMT,with core targets being IL6,TNF and EGFR.Functional enrichment analysis of EMT differential genes showed enrichment in pathways such as neuroactive ligand-receptor interaction,MAPK signaling pathway,endocytosis,calcium signaling pathway,autophagy and PI3K-Akt signaling pathway.Molecular docking showed that IL6,TNF,EGFR bind stably to their corresponding drug ligands.In vitro experiments indicated that Wenjing Decoction could inhibit the PI3K/Akt/mTOR pathway,promote LC3 Ⅰ to LC3 Ⅱ conversion,enhance the expression of Beclin-1,and reduce P62 expression.Moreover,Wenjing Decoction could hinder the expression of the endometriosis-specific biomarker CA125,decrease EGFR,IL-6 and TNF-α expressions in ectopic endothelial cells,inhibiting proliferation.Conclusion Wenjing Decoction can treat EMT through multiple pathways and targets,with the key mechanism being the reversal of autophagy inhibition via down-regulating of the PI3K/Akt/mTOR pathway.

OBJECTIVE: To provide preimplantation genetic testing (PGT) for a patient with Incontinentia pigmenti (IP) due to IKBKG gene variant but without family samples through construction of single nucleotide polymorphism (SNP)-based haplotype by Long-read sequencing (LRS) technology. METHODS: A female IP patient with a heterozygous IKBKG c.1167dup variant but without family genetic data who sought genetic counseling at Women and Children' Hospital of Ningbo University in November 2021 was selected as the study subject. The IKBKG gene has a highly homologous pseudogene IKBKGP1. Genomic DNA was extracted from peripheral blood samples from the couple, and LRS was used to obtain informative SNP loci flanking the variant locus, enabling the construction of SNP haplotype with a long segment spanning from the non-homologous region of IKBKG to the variant site. Trophoblast cells were biopsied from blastocysts fertilized through intracytoplasmic sperm injection, and next-generation sequencing (NGS) was used to determine the SNP information of the embryos. Linkage analysis with the parental SNP haplotypes was conducted to detect the carrier status of the embryos and exclude chromosomal aneuploidies. Sanger sequencing was carried out to validate the result. A euploid embryo without the pathogenic variant was selected for transfer. Prenatal diagnosis was carried out by amniocentesis at mid-trimester to verify the result of PGT tests, and follow-up was conducted after the baby was born. This study has been approved by the Ethics Committee of Women and Children's Hospital of Ningbo University (Ethics No. EC2023-094). RESULTS: A total of seven blastocysts were tested, and PGT results indicated that two embryos were euploid and did not carry the pathogenic variant. One euploid embryo was transferred, which resulted in a singleton pregnancy. Amniocentesis at 24 weeks of gestation confirmed that the status of fetal IKBKG gene, and its chromosomal status was consistent with the PGT results. A healthy male infant was born at 38+6 weeks of gestation. CONCLUSION For IP patients with de novo mutation or without family genetic samples, PGT with LRS can directly construct the SNP-based haplotype while avoiding interference from pseudogenes, providing an effective strategy for PGT.
Female ; Humans ; Male ; Pregnancy ; Genetic Testing/methods* ; Haplotypes/genetics* ; High-Throughput Nucleotide Sequencing/methods* ; I-kappa B Kinase/genetics* ; Incontinentia Pigmenti/diagnosis* ; Polymorphism, Single Nucleotide/genetics* ; Preimplantation Diagnosis/methods* ; Infant, Newborn

OBJECTIVE: To investigate the clinical phenotypes and genetic variant characteristics in children with epilepsy. METHODS: A total of 91 children with epilepsy admitted to the Women's and Children's Hospital Affiliated to Ningbo University from July 2021 to October 2022 were selected as the study subjects. Peripheral blood samples were collected from the children for whole exome sequencing. Candidate genetic variants were validated by Sanger sequencing and copy number variation sequencing (CNV-seq). The clinical phenotypes and treatment outcomes of the children with epilepsy were followed up, and an analysis of the relationship between genotype and phenotype was conducted. This study was approved by the Women's and Children's Hospital Affiliated to Ningbo University (Ethics No.: EC2020-048). RESULTS: Among the 91 children with epilepsy, 21 cases (23.08%, 21/91) were found to carry pathogenic or likely pathogenic variants. Of these, 18 cases had involved single base variant or insertional deletion, while 3 cases involved copy number variations. The gene with the highest detection rate was PRRT2 (38.10%, 8/21). Among the children with genetic variants, 47.62% (10/21) had onset during infancy, with 8 diagnosed with Benign familial infantile epilepsy (BFIE), 8 with Developmental epileptic encephalopathy (DEE), and 3 with Epileptic encephalopathy (EE). One case of Dravet syndrome (DS) and one case of Infantile spasms (IS) were also noted. The clinical manifestations of children were diverse and primarily included generalized tonic-clonic seizures and focal seizures. Among them, 52.38% (11/21) had exhibited cluster seizures, 23.81% (5/21) showed fever sensitivity, and 14.29% (3/21) experienced status epilepticus. After pharmacological treatment, 42.86% (9/21) of children had achieved complete seizure control, while 61.90% (13/21) had intellectual disability and 19.05% (4/21) had co-morbid autism spectrum disorder. CONCLUSION Pathogenic or likely pathogenic variants were identified in 23.08% of the pediatric epilepsy cases, with the PRRT2 gene being the most frequently involved. Among children carrying genetic variants, 47.62% had seizure onset during infancy. Genetic factors are an important cause of epilepsy, and early genetic testing may facilitate precise diagnosis, treatment, and prognostic evaluation.
Humans ; Female ; Male ; Epilepsy/genetics* ; Child, Preschool ; Child ; Phenotype ; Genotype ; DNA Copy Number Variations/genetics* ; Infant ; Membrane Proteins/genetics* ; Nerve Tissue Proteins/genetics* ; Adolescent ; Exome Sequencing

Objective To analyze the correlations between the thoracic electrical impedance tomography(EIT)-derived parameters global inhomogeneity(GI),center of ventilation(COV),regional ventilation delay(RVD),and atelectasis in hospitalized patients,and to explore their effectiveness in evaluating atelectasis.Methods The clinical data of 140 hospitalized patients monitored by thoracic EIT between Sep.2024 and Jan.2025 were retrospectively analyzed.Patients were assigned to 2 groups based on chest computed tomography confirmation of atelectasis within the preceding short-term period during EIT monitoring:non-atelectasis group or atelectasis group.The algorithm software designed with MATLAB was used to acquire GI and COV.RVD was obtained through analysis with the Dr?ger EIT Data Analysis Tool 6.3 software,and patients'general data were concurrently documented.Comparative analysis of EIT-derived parameters between groups was conducted.Multivariate logistic regression analysis was employed to investigate the correlations of GI,COV,and RVD with atelectasis,while receiver operating characteristic curve analysis was performed to assess the efficacy of EIT-derived parameters in evaluating atelectasis.Results A total of 140 patients were enrolled,with 19(13.6%)cases presenting atelectasis.Compared to the non-atelectasis group,the atelectasis group demonstrated significantly higher proportions of male patients and cardiovascular disease and thoracic surgery(non-pulmonary)histories,lower body mass index(BMI),and alongside elevated GI and RVD values with reduced COV(all P＜0.05).Multivariate logistic regression analysis revealed that GI,COV,and RVD maintained independent associations with atelectasis after adjusting for age,gender,BMI,pleural effusion,and emphysema(odds ratio[OR]=1.39,95%confidence interval[CI]1.20-1.67;OR=0.85,95%CI 0.75-0.96;OR=1.22,95%CI 1.09-1.39;all P＜0.05).The area under curve(AUC)values of GI,COV,and RVD for evaluating atelectasis in hospitalized patients were 0.82,0.80,and 0.82,respectively(while RVD demonstrated a higher AUC,its clinical applicability was influenced by respiratory patterns).Conclusion Thoracic EIT-derived parameters GI and COV demonstrate significant correlations with atelectasis and may serve as valuable indicators for evaluating atelectasis in hospitalized patients.

Objective To investigate the multi-target mechanisms of quercetin in treating endometriosis(EMT)through integrative network pharmacology analysis.Methods Active targets of quercetin were collected from the TCMSP database,while EMT-related differentially expressed genes(DEGs)were identified through the Gene Expression Omnibus(GEO)dataset.A comparative analysis was conducted to pinpoint potential therapeutic targets of quercetin for EMT treatment.Functional enrichment analyses were employed to investigate the biological functions associated with these targets,and a protein-protein interaction(PPI)network was conducted to identify core targets.Molecular docking and dynamics simulations were performed to validate the binding characteristics between quercetin and the core targets.The top 2 target protein pairs,HSP90AB1 and AR,exhibiting the lowest binding energy,were selected for subsequent cellular experimental validation.Human EMT-immortalized ectopic endometrial epithelial cell line 12Z(n=6,independent replicates)was subjected,and CCK-8 assay was used to determine ehe effects of quercetin on cell viability and proliferation,and the half-maximal inhibitory concentration(IC50)was calculated at 48 h after treatment.Then the 12Z cells were treated with quercetin at a concentration gradient of 0,30,60 and 90 μmol/L,the migration and invasion abilities were assessed with cell scratch and cell invasion assays.Western blotting was conducted to detect the changes in the expression of HSP90AB1 and AR proteins after different doses of treatment.Results There were 49 potential EMT-related therapeutic targets and 10 core targets identified.Functional enrichment analyses revealed that these targets were significant enriched in inflammation-related signaling pathways,including AGE-RAGE,ErbB and TNF;immune-related pathways,such as Th17 cell differentiation,T/B cell receptor signaling;angiogenesis-related pathways like VEGF;and hormonal regulatory pathways involving estrogen and GnRH.Molecular docking demonstrated that quercetin exhibited favorable binding activity(binding energy<-5 kcal/mol)with all core target proteins,with particularly strong binding energies(<-7 kcal/mol)observed for AR,EGFR,FOS,ERBB2,and HSP90AB1.Molecular dynamics simulations revealed that quercetin forms sustained hydrogen bond interactions with AR and HSP90AB1,facilitating the formation of stable complexes.CCK-8 assay,cell scratch assay,and transwell invasion assay indicated that quercetin inhibited the proliferative activity,and migrative and invasive abilities of 12Z cells in a concentration-dependent manner,with more pronounced inhibitory effects observed at 60 and 90 μmol/L quercetin(P<0.001);Western blotting revealed that treatment of 12Z cells with varying quercetin concentrations for 48 h up-regulated the expression of HSP90AB1 and AR,with the most significant increase observed at 90 μmol/L quercetin(HSP90AB1,P<0.05;AR,P<0.001).The restored expression levels of HSP90AB1 and AR showed positive correlations with the proliferative activity,migrative and invasive abilities of ectopic endometrial cells.Conclusion Quercetin effectively addresses endometriosis through multiple molecular targets and signaling pathways,and stabilization of the HSP90AB1/AR complex and subsequent protein upregulation represents a key therapeutic mechanism.

Objective To investigate the renal function recovery and perioperative risk factors for chronic kidney disease in patients after acute Stanford type A aortic dissection (ATAAD) repair. Methods A retrospective study was conducted on patients who underwent ATAAD repair at the Xiamen Cardiovascular Hospital, Xiamen University from 2020 to 2021, and their clinical data were analyzed. Results A total of 255 patients were included, with 200 males and 55 females, and an average age of (52.80±12.46) years. The incidence of acute kidney injury (AKI) after ATAAD repair was 43.9%. Dissection involving the renal artery [OR=2.144, 95%CI (1.234, 3.765), P=0.007], intraoperative urine output [OR=0.761, 95%CI (0.625, 0.911), P=0.004], and intraoperative red blood cell transfusion [OR=1.288, 95%CI (1.088, 1.543), P=0.004] were significantly associated with early AKI after ATAAD repair. Long-term renal function follow-up data were available for 232 patients, among whom 40 (17.2%) patients developed chronic kidney disease (CKD). Independent predictors for CKD included lower body mass index [OR=0.827, 95%CI (0.723, 0.931), P=0.003], preoperative cardiac tamponade [OR=5.344, 95%CI (1.65, 17.958), P=0.005], preoperative renal hypoperfusion syndrome [OR=12.629, 95%CI (5.003, 35.373), P<0.001], postoperative peak serum creatinine time>3 d [OR=7.566, 95%CI (2.799, 22.731), P<0.001], and AKI grade [grade 1: OR=4.418, 95%CI (1.339, 15.361), P=0.016; grade 2: OR=8.345, 95%CI (1.762, 40.499), P=0.007; grade 3: OR=9.463, 95%CI (2.602, 37.693), P<0.001]. Conclusion AKI related to ATAAD repair can recover in the early postoperative period, but both the duration and severity of AKI will affect long-term renal function. In addition, patients' nutritional status, preoperative cardiac tamponade, and renal hypoperfusion syndrome are also independent risk factors for long-term renal dysfunction.

BACKGROUND: Spicy food consumption has been reported to be inversely associated with mortality from multiple diseases. However, the effect of spicy food intake on the incidence of vascular diseases in the Chinese population remains unclear. This study was conducted to explore this association. METHODS: This study was performed using the large-scale China Kadoorie Biobank (CKB) prospective cohort of 486,335 participants. The primary outcomes were vascular disease, ischemic heart disease (IHD), major coronary events (MCEs), cerebrovascular disease, stroke, and non-stroke cerebrovascular disease. A Cox proportional hazards regression model was used to assess the association between spicy food consumption and incident vascular diseases. Subgroup analysis was also performed to evaluate the heterogeneity of the association between spicy food consumption and the risk of vascular disease stratified by several basic characteristics. In addition, the joint effects of spicy food consumption and the healthy lifestyle score on the risk of vascular disease were also evaluated, and sensitivity analyses were performed to assess the reliability of the association results. RESULTS: During a median follow-up time of 12.1 years, a total of 136,125 patients with vascular disease, 46,689 patients with IHD, 10,097 patients with MCEs, 80,114 patients with cerebrovascular disease, 56,726 patients with stroke, and 40,098 patients with non-stroke cerebrovascular disease were identified. Participants who consumed spicy food 1-2 days/week (hazard ratio [HR] = 0.95, 95% confidence interval [95% CI] = [0.93, 0.97], P <0.001), 3-5 days/week (HR = 0.96, 95% CI = [0.94, 0.99], P = 0.003), and 6-7 days/week (HR = 0.97, 95% CI = [0.95, 0.99], P = 0.002) had a significantly lower risk of vascular disease than those who consumed spicy food less than once a week ( Ptrend <0.001), especially in those who were younger and living in rural areas. Notably, the disease-based subgroup analysis indicated that the inverse associations remained in IHD ( Ptrend = 0.011) and MCEs ( Ptrend = 0.002) risk. Intriguingly, there was an interaction effect between spicy food consumption and the healthy lifestyle score on the risk of IHD ( Pinteraction = 0.037). CONCLUSIONS Our findings support an inverse association between spicy food consumption and vascular disease in the Chinese population, which may provide additional dietary guidance for the prevention of vascular diseases.
Humans ; Male ; Female ; Prospective Studies ; Middle Aged ; Aged ; Vascular Diseases/etiology* ; Risk Factors ; China/epidemiology* ; Adult ; Proportional Hazards Models ; Cerebrovascular Disorders/epidemiology* ; East Asian People

The disturbance of the human microbiota influences the occurrence and progression of many diseases. Live therapeutic bacteria, with their genetic manipulability, anaerobic tendencies, and immunomodulatory properties, are emerging as promising therapeutic agents. However, their clinical applications face challenges in maintaining activity and achieving precise spatiotemporal release, particularly in the harsh gastrointestinal environment. This review highlights the innovative bacterial functionalized encapsulation strategies developed through advances in physicochemical and biological techniques. We comprehensively review how bacterial encapsulation strategies can be used to provide physical barriers and enhanced adhesion properties to live microorganisms, while introducing superior material properties to live bacteria. In addition, this review outlines how bacterial surface coating can facilitate targeted delivery and precise spatiotemporal release of live bacteria. Furthermore, it elucidates their potential applications for treating different diseases, along with critical perspectives on challenges in clinical translation. This review comprehensively analyzes the connection between functionalized bacterial encapsulation and innovative biomedical applications, providing a theoretical reference for the development of next-generation bacterial therapies.