Objective: To define the mechanism by which swine TRIM25 restricts vesicular stomatitis virus replication. Methods: Porcine 3D4/21 cells with TRIM25 overexpression or knockdown were infected with vesicular stomatitis virus. Viral replication was quantified by immunoblotting, quantitative reverse transcription polymerase chain reaction, and 50% tissue culture infectious dose assays. Type I interferon signaling was assessed by transcript quantification, interferon-beta and interferon-stimulated response element reporter assays, and co-immunoprecipitation.Viral RNA binding was tested by RNA immunoprecipitation. Results: TRIM25 overexpression reduced viral RNA and infectious titers, whereas TRIM25 knockdown increased replication (p < 0.01). TRIM25 increased interferon-beta and interferon-stimulated gene expression and enhanced interferon-beta and interferonstimulated response element promoter activity (p < 0.01). Mechanistically, TRIM25 promoted Lys63-linked ubiquitination of RIG-I and increased phosphorylation of TANK-binding kinase 1 and interferon regulatory factor 3. TRIM25 also bound vesicular stomatitis virus genomic RNA, and binding required the C-terminal region. Conclusions and Relevance: Porcine TRIM25 restricts vesicular stomatitis virus replication by amplifying type I interferon signaling and directly binding viral RNA.

Objective To evaluate the efficacy and safety of a combination therapy regimen comprising stereotactic body radiation therapy (SBRT) combined with liposomal irinotecan, camrelizumab, and anti-angiogenic therapy (CAP) in patients with advanced bone and soft tissue sarcomas. Methods This study is a prospective subgroup analysis of the multicenter, open-label, single-arm phase Ⅱ CAP trial (NCT04569916), which enrolled 60 patients with advanced solid tumors who had progressed after standard therapy between November 2020 and February 2023, including 11 patients with advanced bone and soft tissue sarcomas. All patients received sequential liposomal irinotecan within 48 hours after stereotactic body radiotherapy (SBRT), followed by maintenance therapy with camrelizumab and an anti-angiogenic agent until disease progression or intolerable toxicity. The primary efficacy endpoint was objective response rate (ORR). Secondary endpoints included disease control rate (DCR), progression-free survival (PFS), overall survival (OS), and safety. Results In the bone and soft tissue sarcoma subgroup (n=11), three patients achieved partial response (PR), five had stable disease (SD), and three experienced disease progression (PD). ORR was 27.3%, and DCR was 72.7%. mPFS was 5.8 months (95%CI: 2.2-10.3), and mOS not reached. Regarding safety, adverse reactions primarily included bone marrow suppression, fatigue, and gastrointestinal reactions, indicating overall manageable toxicity. Conclusion As a subgroup analysis of the CAP study, this research suggests that radiotherapy combined with liposomal irinotecan, camrelizumab, and anti-angiogenic therapy demonstrates certain efficacy in advanced bone and soft tissue sarcomas that have failed multiple lines of treatment, with overall manageable adverse reactions. Future studies should expand the sample size.

OBJECTIVE: To explore the potential effects and mechanisms of Liang-Ge-San (LGS) for the treatment of acute respiratory distress syndrome (ARDS) through network pharmacology analysis and to verify LGS activity through biological experiments. METHODS: The key ingredients of LGS and related targets were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform. ARDS-related targets were selected from GeneCards and DisGeNET databases. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed using the Metascape Database. Molecular docking analysis was used to confirm the binding affinity of the core compounds with key therapeutic targets. Finally, the effects of LGS on key signaling pathways and biological processes were determined by in vitro and in vivo experiments. RESULTS: A total of LGS-related targets and 496 ARDS-related targets were obtained from the databases. Network pharmacological analysis suggested that LGS could treat ARDS based on the following information: LGS ingredients luteolin, wogonin, and baicalein may be potential candidate agents. Mitogen-activated protein kinase 14 (MAPK14), recombinant V-Rel reticuloendotheliosis viral oncogene homolog A (RELA), and tumor necrosis factor alpha (TNF-α) may be potential therapeutic targets. Reactive oxygen species metabolic process and the apoptotic signaling pathway were the main biological processes. The p38MAPK/NF-κ B signaling pathway might be the key signaling pathway activated by LGS against ARDS. Moreover, molecular docking demonstrated that luteolin, wogonin, and baicalein had a good binding affinity with MAPK14, RELA, and TNF α. In vitro experiments, LGS inhibited the expression and entry of p38 and p65 into the nucleation in human bronchial epithelial cells (HBE) cells induced by LPS, inhibited the inflammatory response and oxidative stress response, and inhibited HBE cell apoptosis (P<0.05 or P<0.01). In vivo experiments, LGS improved lung injury caused by ligation and puncture, reduced inflammatory responses, and inhibited the activation of p38MAPK and p65 (P<0.05 or P<0.01). CONCLUSION LGS could reduce reactive oxygen species and inflammatory cytokine production by inhibiting p38MAPK/NF-κ B signaling pathway, thus reducing apoptosis and attenuating ARDS.
Drugs, Chinese Herbal/pharmacology* ; Respiratory Distress Syndrome/enzymology* ; p38 Mitogen-Activated Protein Kinases/metabolism* ; NF-kappa B/metabolism* ; Animals ; Signal Transduction/drug effects* ; Molecular Docking Simulation ; Humans ; Male ; Network Pharmacology ; Apoptosis/drug effects* ; Mice

Hearing loss is the most prevalent disabling disease. Cochlear implantation（CI） serves as the primary intervention for severe to profound hearing loss. This consensus systematically explores the value of genetic diagnosis in the pre-operative assessment and efficacy prognosis for CI. Drawing upon domestic and international research and clinical experience, it proposes an evidence-based medicine three-tiered prognostic classification system（Favorable, Marginal, Poor）. The consensus focuses on common hereditary non-syndromic hearing loss（such as that caused by mutations in genes like GJB2, SLC26A4, OTOF, LOXHD1） and syndromic hereditary hearing loss（such as Jervell & Lange-Nielsen syndrome and Waardenburg syndrome）, which are closely associated with congenital hearing loss, analyzing the impact of their pathological mechanisms on CI outcomes. The consensus provides recommendations based on multiple round of expert discussion and voting. It emphasizes that genetic diagnosis can optimize patient selection, predict prognosis, guide post-operative rehabilitation, offer stratified management strategies for patients with different genotypes, and advance the application of precision medicine in the field of CI.
Humans ; Cochlear Implantation ; Prognosis ; Hearing Loss/surgery* ; Consensus ; Connexin 26 ; Mutation ; Sulfate Transporters ; Connexins/genetics*

BackgroundExecutive function deficits constitute a core problem in attention-deficit/hyperactivity disorder （ADHD）. Previous assessments of executive function in children with ADHD have predominantly relied on performance-based neuropsychological tests conducted in laboratory settings， though their predictive validity for real-world functional outcomes remains limited. In contrast， ecological executive function emphasizes the evaluation of complex task management in naturalistic contexts， demonstrating a stronger predictive power for functional adaptation in daily living among children with ADHD， such as multitasking performance， social interactions and so on. However， current empirical evidence regarding ecological executive function in this population remains insufficient. ObjectiveTo investigate the executive function characteristics of children with ADHD from an ecological perspective， thereby providing references for developing targeted interventions. MethodsA case control study was conducted， including 277 ADHD children who met the Diagnostic and Statistical Manual of Mental Disorders， fourth edition （DSM-IV） criteria and were selected at the Child Health Care and Mental Health Center of Shenzhen Children's Hospital from June 2017 to December 2020， as well as 98 healthy controls were recruited from primary and secondary schools in Shenzhen. All participants were assessed using Wechsler Intelligence Scale for Children， fourth edition （WISC-IV） and Behavior Rating Inventory of Executive Function （BRIEF）. Differences in WISC-IV and BRIEF scores were compared between ADHD group and control groups， followed by the comparison of BRIEF scores by gender and ADHD subtypes. ResultsAmong the 277 children with ADHD， 136 cases （49.10%） had predominantly inattentive type （ADHD-I）， 6 cases （2.17%） had predominantly hyperactive-impulsive type （ADHD-HI）， and 135 cases （48.73%） had combined type （ADHD-C）. ADHD group demonstrated significantly lower scores on both the WISC-IV total IQ and four index scores （verbal comprehension， perceptual reasoning， working memory and processing speed） than control group （t=3.698~9.335， P<0.01）. After controlling for WISC-IV total IQ as a covariate， the scores of each factor in the dimensions of behavioral regulation index （inhibition， shifting， emotional control） and metacognition index （task initiation， working memory， planning， monitoring and organization） were all higher in ADHD group than in control group， and the differences were statistically significant （F=46.563~290.475， P<0.01）. In terms of gender， no statistically significant difference was found in BRIEF composite scores （behavioral regulation index or metacognition index） of children with ADHD （t=0.105~1.190， P>0.05）. In terms of ADHD subtypes， children with ADHD-C reported significantly higher scores than those with ADHD-I on the scores of inhibition， emotional control， organization and monitoring in BRIEF （t=2.481~7.343， P<0.05 or 0.01）. ConclusionChildren with ADHD have multidimensional deficits in ecological executive function， which vary across different subtypes. ［Funded by Shenzhen Excellent Science and Technology Innovation Talent Training Project （number， RCYX20221008092849069）； the Sanming Project of Medicine in Shenzhen］

Objective To investigate the mechanism by which Senegenin(SEN)alleviates microglial inflammatory response through the nuclear factor erythroid 2-related factor 2(Nrf2)/NOD-like receptor protein 3(NLRP3)pathway.Methods BV2 mouse microglia cells were randomly divided into control group,model group,SEN group and MCC950 group.Cells in control group were not treated,and cells in model group were added with 1 μg/ml lipopolysaccharide(LPS);Cells in SEN group were added with 1 μg/ml LPS+4 μmol/L SEN,and cells in MCC950 group were added with 1 μg/ml LPS+10 μmol/L MCC950 for 24 hours.CCK-8 method was used to detect the effect of different concentrations of SEN on the viability of BV2 cells.Griess method was used to determine the release amount of nitric oxide(NO)in the supernatant.Real-time fluorescent quantitative PCR was used to determine the mRNA expression levels of NLRP3,lymphocyte apoptosis-associated spect-like protein containing a CARD(ASC),caspase-1,interleukin(IL)-1β and IL-18 mRNA.Immunofluorescence staining was used to detect the expression levels of ASC,IL-1β,Nrf2 and heme oxygenase-1(HO-1).Western blotting was used to detect the expression levels of NLRP3,caspase-1,ASC,IL-1β,IL-18,Nrf2,HO-1,nuclear factor kappa B(NF-κB)and inducible nitric oxide synthase(iNOS).Results The results of CCK-8 method showed that there was no significant difference in the viability of BV2 cells treated with 2～20 μmol/L SEN compared with control group(P>0.05).Compared with control group,the viability of BV2 cells in model group decreased significantly(P<0.05).Compared with model group,the viability of BV2 cells in 4 μmol/L SEN group was significantly restored(P<0.05).Compared with control group,the results of Griess method showed that the release amount of NO in cells of model group increased significantly(P<0.05);the results of real-time PCR showed that the expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 mRNA in cells of model group increased significantly(P<0.05);the results of Western blotting showed that the protein expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 proteins in cells of model group increased significantly(P<0.05),and the immunofluorescence staining results showed that the expression levels of iNOS and NF-κB protein in cells of model group increased,and the expression levels of Nrf2 and HO-1 decreased,with statistically significant differences(P<0.05).Compared with model group,the release amount of NO in cells of SEN group and MCC950 group decreased,and the expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 mRNA and proteins decreased,with statistically significant differences(P<0.05);in the SEN group,the expression levels of iNOS and NF-κB decreased,and immunofluorescence staining showed that Nrf2 was translocated into the nucleus,and the expression levels of Nrf2 and HO-1 proteins increased significantly,with statistically significant differences(P<0.05).Conclusions SEN could alleviate the inflammatory response of mouse microglia cells induced by LPS and inhibit the activation and expression of NLRP3 inflammasome,with an effect comparable to that of the inflammasome inhibitor MCC950.The mechanism may be related to the regulation of the expression of upstream factors Nrf2 and HO-1.

Objective To explore the regulatory effect of amylin on the feeding behavior of mice through the ERK pathway of paraventricular nucleus of the hypothalamus(PVN).Methods A total of 54 C57BL/6J mice were randomly divided into multiple groups for experiments.First,immunofluorescence was used to detect the distribution of ERK and calcitonin receptor(CTR)neurons in the brains of 4 mice.A chemical lesion model of PVN neurons was established,and 16 mice were divided into four groups(4 mice per group):control(Ctrl)group,ibotenic acid(IBO)-lesioned group,amylin group,and amylin+IBO group.The number of ERK-positive neurons in the PVN region and food intake were then measured.Subsequently,16 mice were divided into four groups(4 mice per group),Ctrl group,amylin group,ERK antagonist U0126 group,and amylin+U0126 group.The expression level of the immediate early gene c-fos in the PVN was detected by Real-time PCR.Immunofluorescence was used to detect the distribution of p-ERK positive neurons in the brains of 4 saline-treated mice and 4 amylin-treated mice.Finally,18 mice were randomly divided into three groups(6 mice per group)through intracerebroventricular injection,Ctrl group,amylin group,and amylin+receptor antagonist AC 187 group.Then,triple immunofluorescence was used to detect the co-expression of p-ERK,neuronal nitric oxide synthase(nNOS),and CTR-immunopositive neurons in the PVN region,and food intake within 3 hours after treatment was recorded simultaneously.Results ERK-positive neurons were mainly distributed in the suprachiasmatic nucleus(SCN),arcuate nucleus(ARC),and locus coeruleus(LC),while ERK and CTR co-expressing positive neurons were specifically enriched in the PVN(P=0.0012).Lesion of PVN neurons led to a decrease in the number of ERK-positive neurons(P＜0.001)and blocked the anorectic effect of amylin(P＜0.001).Amylin activated the expression of the c-fos gene in the PVN region(P＜0.05),and this effect was inhibited by U0126(P＜0.05).After intracerebroventricular injection of amylin,the number of triple-positive neurons for p-ERK,nNOS,and CTR in the PVN increased(P＜0.05).Addition of AC 187 blocked the activation effect of amylin(P＜0.01)and reversed the anorectic effect of amylin(1 h,P=0.2218;2 h,P=0.2218;3 h,P=0.6974).Conclusion P-ERK neurons in the PVN regulate feeding behavior through an amylin receptor-dependent ERK signaling pathway.

Objective This study aimed to investigate natural infection of rodents with Ehrlichia and Neoehrlichia at major Chinese land-border ports along the"Belt and Road".Methods In 2022,rodents were monitored in 10 ports in northern and southern China and identified based on diagnostic morphological characteristics.The 16S rRNA genes of Ehrlichia and Neoehrlichia were detected by PCR using universal primers from rodent samples and phylogenetic analysis was performed based on the sequences of the detected positive pathogens.Results A total of 356 rodents were sampled,including 2 orders,5 families,15 genera,and 20 species.Predominantly,73,61,56,and 58 were Meriones unguiculatus(20.51%),Rattus norvegicus(17.13%),Apodemus agrarius(15.73%),and Microtus gregalis(16.29%).Only one Microtus fortis from Suifenghe Port was infected with Ehrlichia sp.Moreover,12 rodents were infected with Neoehrlichia spp.(overall positivity rate:3.37%).Conclusions Natural infections with Ehrlichia spp.and Neoehrlichia spp.were demonstrated in rodents at important Chinese land-border ports.The positivity rate of Neoehrlichia spp.was high in some ports,indicating that surveillance for ticks and their prevention and control measures should be intensified in these regions.

Objective:To explore the genetic characteristics of a fetus affected with Structural heart defects and renal anomalies syndrome (SHDRA).Methods:A pedigree with SHDRA (fetus and the parents) who had visited the Affiliated Women and Children′s Hospital of Ningbo University in April 2023 was selected as the study subject. Clinical data of the family were collected. A total of 10 mL of amniotic fluid cells from the fetus and 5 mL of peripheral blood samples from the parents were collected for genomic DNA extraction. Trio whole-exome sequencing (Trio-WES) was performed, and Sanger sequencing was used to validate candidate variants in the family. The identified variants were classified according to the Standards and Guidelines for the Interpretation of Sequence Variants established by the American College of Medical Genetics and Genomics (ACMG) (hereinafter referred to as the " ACMG Guidelines). Relevant research literature on SHDRA in domestic and international databases were searched for literature review. This study was approved by the Affiliated Women and Children′s Hospital of Ningbo University (Ethics No. EC2023-094).Results:①In this family, prenatal ultrasound at 18 weeks of gestation revealed left renal multicystic dysplasia in the fetus. After birth, the infant exhibited an ostium secundum atrial septal defect, patent ductus arteriosus, and left renal multicystic dysplasia. Trio-WES revealed that the fetus had carried c. 344dup(p.L116Afs*32) and c. 90_104dup(p.Ala31_Ala35dup) compound heterozygous variants in the TMEM260 gene, which were respectively inherited from its father and mother. According to the ACMG guidelines, the c. 344dup(p.L116Afs*32) and c. 90_104dup (p.Ala31_Ala35dup) variants were classified as pathogenic (PM2_Supporting+ PVS1+ PP4) and likely pathogenic (PM2_Supporting+ PM4+ PM3+ PP4), respectively. ②According to the literature search strategy set for this study, a total of 6 literature was retrieved, involving 25 SHDRA patients from 20 families. Together with the patients in this study, there were 14 TMEM260 gene variants, most of which were frameshift variants (7 types) and had located in exons 3, 11 and 13. The main clinical features of SHDRA were congenital heart malformation, renal abnormality and neurodevelopmental abnormality, and there was a lack of genotype-phenotype correlation. Conclusion:The c. 344dup(p.L116Afs*32) and c. 90_104dup(p.Ala31_Ala35dup) variants of the TMEM260 gene probably underlay the SHDRA in this family. Above finding has provided a basis for clinical diagnosis and genetic counseling for the family.

Objective:To assess the effects of various leisure hobbies on the ability of daily living(ADL)and the quality of life among older adults in China.Methods:A cross-sectional study was conducted using data from the 4th Chinese Urban and Rural Elderly Living Conditions Sample Survey.We categorized community recreation hobbies into two types: dynamic hobbies, which include walking, playing ball games, dancing, and other physical activities, and static hobbies, which encompass activities that do not require physical exertion, such as reading, watching TV, and drawing.The EuroQol five-dimension scale(EQ-5D)-3L was employed to evaluate the quality of life.Older adults who maintained either dynamic or static hobbies for three consecutive years from 2017 to 2019 were initially screened.The relationship between different types of leisure hobbies and ADL scores, as well as quality of life at various time points, was analyzed using repeated-measures ANOVA.Subsequently, propensity scores were matched based on gender, age, ethnicity, marital status, literacy, and chronic diseases, and five machine learning algorithms were utilized to identify the optimal model for predicting fully independent ADL and EQ-5D effect values.Based on the optimal predictive model, we conducted Shapley Additive Explanations(SHAP)and global analysis.Results:The total number of older adults who participated in the survey over the three consecutive years from 2017 to 2019 was 4 912.Among these, 2 594 individuals maintained the same hobby habits throughout this period, comprising 1 362 males and 1 232 females, with an average age of(70.81±6.71)years.Statistically significant differences in ADL scores and EQ-5D scores were observed between the dynamic and static leisure hobby groups(all P<0.05)across various time points, hobby group classifications, and interaction levels.Older adults engaged in dynamic hobbies exhibited superior ADL functioning and higher EQ-5D scores in comparison to those in the static hobby group, with these differences remaining statistically significant following propensity score matching(all P<0.05).In predicting full ADL independence, the SHAP value for dynamic hobbies ranked second, while for predicting an EQ-5D weighted value of 1, the SHAP value for dynamic hobbies ranked ninth.Overall, the type of dynamic hobby demonstrated a positive trend in its effect on the ability to perform daily activities independently and in achieving an EQ-5D weighted value of 1. Conclusions:In comparison to static hobbies, Chinese older adults who engage in dynamic hobbies exhibit improved daily mobility and a higher quality of life.Furthermore, dynamic hobbies positively influence the capacity for full independence in daily activities, as evidenced by an EQ-5D weighted value of 1.