Background Carbendazim (CBZ), a widely used benzimidazole fungicide, has raised increasing concerns regarding the health risks associated with its residues. However, the toxic effects and associated mechanisms of CBZ on the skeletal system have not been reported. Objective To elucidate the effects of carbendazim on osteogenic differentiation and its underlying mechanisms. Methods MC3T3-E1 mouse pre-osteoblastic cells were treated with 1, 10, and 100 μmol·L⁻¹ CBZ for 24 h to examine cell viability, alkaline phosphatase (ALP) activity, bone nodule formation, reactive oxygen species (ROS) level, malondialdehyde (MDA) content, and nitric oxide synthase (NOS) activity. Transcriptomics was used to identify differentially expressed genes (DEGs) in osteoblasts exposed to CBZ. Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene set enrichment analysis (GSEA) were employed to analyze the potential biological pathways of DEGs. Real-time polymerase chain reaction (RT-PCR) and Western blot were used to validate changes in gene and protein expression. Results Exposure to 10 and 100 μmol·L⁻¹ CBZ significantly reduced osteoblast viability, ALP activity, bone nodule formation, and NOS activity, while increasing intracellular ROS levels. CBZ at 100 μmol·L⁻¹ concentration significantly elevated MDA level (P < 0.05). The transcriptomic analysis revealed that 1 μmol·L⁻¹ CBZ treatment resulted in 385 significantly DEGs. The KEGG enrichment analysis revealed that CBZ significantly affects hormone regulation pathways (including parathyroid hormone, growth hormone, dopamine, and oxytocin), mitogen-activated protein kinase (MAPK) and cyclic GMP-dependent protein kinase G (cGMP-PKG) signaling pathways, focal adhesion and adherens junction, as well as the NOD-like receptor signaling pathway and the mRNA surveillance (NMD) pathway. The results of GSEA showed that CBZ significantly inhibited the bile acid metabolism and the Wnt/β-catenin pathway in osteoblasts. The validation results demonstrated that CBZ significantly suppressed the mRNA expression of Wnt3a and β-catenin, as well as the protein expression of Runx2 and Osterix in the Wnt/β-catenin pathway. Conclusion CBZ exposure exhibits potential skeletal toxicity, and its mechanism is through promoting oxidative stress, interfering with the Wnt/β-catenin pathway in osteogenic differentiation, thereby inhibiting the bone formation function of osteoblasts.

ObjectiveTo investigate the effects of metformin on the immune functions of immature dendritic cells （imDCs） and the underlying mechanisms. MethodsMouse bone marrow-derived imDCs were treated with different concentrations of metformin. The working concentration and treatment time of metformin in this study were determined based on the results of cell apoptosis and cell viability assays. The effects of metformin on the phagocytic capacity of imDCs was evaluated using an antigen endocytosis assay. The expression of cluster of differentiation 205 （CD205）， the polymerization of filamentous actin （F-actin）， and the underlying regulatory mechanisms were investigated through flow cytometry， laser confocal fluorescence microscopy， and Western blot. ResultsThe working concentrations of metformin were 1， 2， 4 mmol/L for 24 h determined by the apoptosis and cell viability assays.Metformin significantly suppressed the phagocytic capacity of imDCs， down-regulated the expression of the mannose receptor CD205 on the cell surface， which was closely associated with phagocytic function； metformin inhibited the RhoA-ROCK1-LIMK1-Cofilin signaling pathway， which inhibited the polymerization of F-actin and disturbed its dynamic remodeling of imDCs. ConclusionMetformin can inhibit the expression of CD205 and disrupt the remodeling of F-actin， thereby suppressing the antigen-capturing capacity of imDCs.

Objective: To evaluate oral health status and knowledge, attitude and practice (KAP) among third grade primary school students after the implementation of the Comprehensive Intervention Project for Childhood Oral Diseases in Guizhou Province, so as to provide evidence for program optimization. Methods: Using a multistage cluster sampling method, oral examinations and questionnaire surveys were conducted among third grade primary school students in schools covered by the Comprehensive Intervention Project for Childhood Oral Diseases in Guizhou Province (intervention group, n =1 030) and schools not covered by the program (control group， n =1 130) in November 2023. A follow up survey was conducted one year after the project s implementation (December 2024) to evaluate the differences in oral health status and KAP between the two groups. The t-test and χ 2 test were used to compare means and rates, respectively. Results: One year after implementation, the permanent caries prevalence, caries filling ratio, mean dmft/DMFT, and mean simplified debris index in the intervention group were 33.3%, 22.3 %, 3.44±3.10, 0.77±0.56, which were all better than those in the control group (39.8%, 17.3%, 3.89±2.85, and 0.87± 0.63 ) ( χ 2/ t =9.87, 8.78, -2.59, -3.91,all P <0.05). After intervention, the levels of oral health knowledge, attitude and behaviors in the intervention group were better than those in the control group ( Z =7.69, 2.35, 2.10, all P <0.05). Except for the item "fluoride has no protective effect on teeth", the correct response rates of other oral health knowledge items in the intervention group were higher than those in the control group ( χ 2=3.92-163.94, all P <0.05). The reported rates of tooth brushing related behaviors and sugary beverage consumption were higher in the intervention group than in the control group ( χ 2=4.02-11.42，all P <0.05). Conclusions The Comprehensive Intervention Project for Childhood Oral Diseases improves children s oral health status, oral health knowledge, and some oral health behaviors, but has limited effects on some behaviors and oral health attitudes. Future efforts should focus on the factors influencing the transformation of children s oral health attitudes and behaviors, along with strengthening long term follow up evaluations.

Objective:To develop and evaluate a nomogram prediction model for poor outcome of lower extremity motor function in patients with acute ischemic stroke (AIS) at 90 days after onset.Methods:Patients with AIS admitted to Guangzhou Province Hospital of Chinese Medicine from January to October 2024 were included retrospectively. At 90 days after onset, Functional Ambulation Category (FAC) was used for outcome evaluation. ≥4 was defined as good outcome and <4 was defined as poor outcome. Multivariate logistic regression analysis was used to identify the independent predictive factors for poor outcome of lower extremity motor function, and develop a nomogram prediction model. The area under the receiver operating characteristic curve, calibration curve, and clinical decision curve were used to evaluate the predictive model. Results:A total of 325 patients with AIS were enrolled, including 214 males (65.8%), median aged 62 years (interquartile range, 54-71 years); 158 patients (48.6%) had poor outcome of lower extremity motor function. Multivariate logistic regression analysis showed that older age (odds ratio [ OR] 1.037, 95% confidence interval [ CI] 1.011-1.065]; P=0.007) and a higher baseline National Institutes of Health Stroke Scale (NIHSS) score ( OR 1.472, 95% CI 1.336-1.637; P<0.001) were the independent predictors of poor outcome, while intravenous thrombolysis ( OR 0.195, 95% CI 0.080-0.443; P<0.001) and early rehabilitation intervention ( OR 0.444, 95% CI 0.231-0.850; P=0.014) were the independent predictors of good outcome. The area under the receiver operating characteristic curve of the nomogram prediction model developed using the above factors was 0.906 (95% CI 0.872-0.940), indicating that the model had good discriminability. The calibration curve fits well with the ideal curve. The clinical decision curve showed that the model had stronger clinical practicality. Conclusion:The nomogram developed by age, intravenous thrombolysis, early rehabilitation intervention, and baseline NIHSS score can effectively predict the risk of poor outcome of lower extremity motor function in patients with AIS and has higher clinical value.

Objective To analyze the research literature on fire needling therapy published on China National Knowledge Infrastructure(CNKI),Wanfang Data Knowledge Service Platform(Wanfang),and China Science and Technology Journal Database(VIP)in the last 20 years,and to explore the current status and the hotspots of the research,thus to summarize and analyze the spectrum of the dominant diseases and technological transformations of fire needling therapy.Methods CiteSpace and VOSviewer were applied to analyze the keywords,authors and institutions,to draw the scientific knowledge map,and to summarize the disease spectrum.Results A total of 4 226 literature were included,from 2000 to 2023,the number of publications on fire needling therapy research showed an increasing trend,with each of the four periods showing its own characteristics of the times.The"filiform fire needling"and"pricking of fire needling"were widely used as acupuncture tools and acupuncture techniques in the studies,the emergence of high-frequency disease keywords such as"shingles""vitiligo"and"acne"indicates that research on the treatment of skin diseases with fire needling has attracted much attention.The disease spectrum of fire needling therapy was enlarged,involving a total of 356 diseases distributed in 20 disease systems.Conclusion Skin diseases and arthralgia became the dominant diseases of fire needling therapy,the rise of research hot words such as"acne vulgaris""vitiligo""stroke""chronic eczema""filiform fire needling"and"Lingnan fire needling"indicates that fire needling therapy is developing towards research standardization,theoretical diversification and disease diversification.

Objective To investigate the mechanisms by which Porphyromonas gingivalis(P.gingivalis)promotes the malignant progression of oral squamous cell carcinoma(OSCC)through the recruitment of chemokine receptor 6-positive(CCR6+)regulatory T cells(Treg)in the tumor microenvironment(TME).Methods The Cancer Genome Atlas(TCGA)database was used to analyze the correlation between chemokine ligand 20(CCL20),CCR6,and Treg.The Treg enrichment index and the expression levels of interleukin(IL)-10 and tumor necrosis factor β1(TGF-β1)were assessed in the high CCR6 expression group of OSCC patients.C57BL/6 mice were randomly assigned to a control group and an experimental group(n=6 in each group).The control group received a single injection of 100 μL SCC7,a mice head and neck squamous carcinoma cell line,while the experimental group received a single injection of 100 μL mixture of SCC7 cells and P.gingivalis in the cheek.After two weeks,the mice were sacrificed,and immunohistochemistry was performed to assess the expression levels of CCR6 and forkhead box protein 3(FOXP3)in OSCC.Flow cytometry was performed to analyze the effects of P.gingivalis on OSCC malignant biological behavior,CCR6+Treg cells,and the immune microenvironment.Results Bioinformatics analysis revealed a correlation between CCL20,CCR6,and Treg(r=0.373,P＜0.0001).OSCC patients with high CCR6 expression showed higher Treg enrichment scores and increased IL-10 expression.Animal experiments showed that P.gingivalis promoted the increase in the tumor volume(mm3)(0.294±0.105 in the control group and 0.526±0.101 in the experimental group,P＜0.01)and mass(mg)(206.200±53.950 in the control group and 376.000±119.200 in the experimental group,P＜0.01)in mice with OSCC.Immunohistochemistry confirmed a correlation between CCR6 and FOXP3(r=0.659,P＜0.05),and P.gingivalis promoted the expression of CCR6 and FOXP3.Flow cytometry analysis showed that P.gingivalis increased the proportion of CCR6+Treg(％)(13.780±1.506 in the control group and 18.260±2.257 in the experimental group,P＜0.01)and decreased the proportion of CD8+T cells(％)(27.120±1.647 in the control group and 21.060±3.148 in the experimental group,P＜0.01)in OSCC,thereby promoting the formation of a immunosuppressive microenvironment.Conclusion P.gingivalis promotes the malignant progression of OSCC by recruiting CCR6+Treg cells to form an immunosuppressive TME.

ObjectiveTo explore the role of flurochloridone (FLC) on osteogenic differentiation and the potential mechanism of inhibiting bone formation, and to provide new insights into bone health risks associated with FLC pesticide exposure. MethodsNeonatal rat skull differentiation primary osteoblast model was used to investigate the effects of 1, 10 and 100 μmol·L^-1 FLC exposure on cell viability, osteogenic differentiation alkaline phosphatase (ALP) activity, and bone mineralization nodule formation, respectively. The potential mechanism underlying the inhibition of FLC on osteoblast differentiation was analyzed using osteogenic differentiation gene chip technique, and the expression of key genes and proteins in the pathway was validated using reverse transcription polymerase chain reaction (RT-PCR) and protein immunoblotting (Western blot) methods. ResultsExposure to FLC at a concentration of 100 μmol·L⁻¹ reduced cell proliferation, ALP activity, and the formation of mineralized nodules in primary osteoblasts. Gene chip analyses revealed that exposure to 10 μmol·L⁻¹ FLC induced 15 differentially expressed genes (DEGs). Among these, MMP9 and Tnf were up-regulated, while Nkx3⁃2, Tuft1, Bmp2, Col12a1, Pparg, Enam, Igf1, Bmp5, Bmp3, Calcr, Egf, Igfbp3, and Col14a1 were down-regulated. Results of protein-protein interaction analyses and gene ontology enrichment analyses indicated that FLC inhibited the BMP/SMAD pathway involved in osteogenic differentiation. FLC suppressed the protein expression of BMP2 and Osterix, as well as the expression of key genes critical for osteogenic differentiation and ossification, such as BMP2, Runx2, SMAD1, and SMAD5 in the BMP/SMAD pathway. ConclusionFLC affects osteogenic differentiation and bone formation potential by regulating the BMP/SMAD axis and the expression of osteogenic genes, suggesting its potential risk in bone metabolism.

OBJECTIVE To explore the clinical efficacy of Wuhuang Shengji decoction irrigation combined with inci-sion and hanging suture in patients with perianal abscess.METHODS A total of 121 patients with perianal abscess admitted to the Anorectal Department of Ningbo Traditional Chinese Medicine Hospital from Jan.2022 to Jan.2024 were selected as the study subjects.According to the treatment methods,they were divided into a control group(n=37),a positive control group(n=41)anda research group(n=43).The control group was treated with incision and hanging suture,the positive control group was treated with recombinant human epidermal growth factor combined with incision and hanging suture,and the research group was treated with Wuhuang Shengji Decoction irrigation combined with incision and hanging suture.The clinical efficacy,the time of necrotic tissue shedding,new skin appearing,wound itch disappearing and wound healing and the recurrence rate were ob-served.The visual analogue scale(VAS),wound secretion score and expression levels of vascular endothelial growth factor(VEGF)and platelet-derived endothelial cell growth factor(PD-ECGF)in wound secretions were compared before and 14 days after treatment in the three groups.RESULTS The clinical efficacy of the study group(90.70%)was higher than that of the positive control group(87.80%)and control group(70.27%);the recur-rence rate of the study group(2.33%)was lower than that of the positive control group(9.76%)and the control group(18.92%),and the differences were statistically significant(P<0.05).The time required for necrotic tissue shedding,new skin appearing,wound itch disappearing and wound healing followed the trend of study group

Postoperative infection of internal fixation of closed fractures the lower limbs in adults represents a devastating complication, characterized by diagnostic challenges, prolonged treatment duration and high disability rates. Current management of these infections faces multiple challenges, such as difficulties in early accurate diagnosis, and various controversies about the treatment plan, leading to poor overall diagnosis and treatment results. To address these issues, based on evidence-based medicine and principles with emphasis on scientific rigor, clinical applicability and innovation, the Trauma Branch of the Chinese Medical Association, Orthopedic Branch of the Chinese Medical Doctor Association, Orthopedics Branch of the Chinese Medical Association, and Trauma Orthopedics and Polytrauma Group of the Resuscitation and Emergency Committee of the Chinese Medical Doctor Association have collaboratively organized a panel of relevant experts to develop the Guideline for diagnosis and treatment of infection after internal fixation of closed lower limb fractures in adults ( version 2025). The guideline proposed 10 recommendations, aiming to provide a foundation for standardized diagnosis and treatment of postoperative infection in adults with closed lower limb fractures.

Objective:To construct multimorbidity patterns among elderly individuals with chronic diseases and to explore the relationship between these patterns and frailty.Methods:A cross-sectional study was conducted involving 4, 706 elderly participants aged 60 years and older from selected prefecture-level cities in Shaanxi Province.Data were collected on general information, chronic diseases, and frailty status.The average age of the participants was 69.9±6.7 years, with males comprising 47.3%(2, 255 cases)and females comprising 52.7%(2, 481 cases)of the sample.Latent class analysis(LCA)was employed to identify multimorbidity patterns, while multivariate logistic regression analysis was utilized to examine the associations between these patterns and frailty.Results:The prevalence of multimorbidity within the study population was found to be 43.6%(2, 052 cases out of 4, 706 cases).The highest rates of multimorbidity were observed in anxiety and depression(100%, 23 cases out of 23 cases), dementia(100%, 6 cases out of 6 cases), and Parkinson's disease(100%, 11 cases out of 11 cases).Stroke followed closely with a rate of 96.8%(597 cases out of 617 cases), while rheumatoid arthritis exhibited the lowest rate of multimorbidity with other chronic diseases at 50%(4 cases out of 8 cases).Five distinct multimorbidity patterns were identified through LCA: the complex multimorbidity class(123 cases), the stroke-respiratory class(546 cases), the sleep disorders-osteoarticular class(488 cases), the cardiovascular-metabolic class(987 cases), and the relatively healthy class(2, 562 cases).When compared to the relatively healthy class, the complex multimorbidity class( OR=2.317, 95% CI: 1.573-3.412), stroke-respiratory class( OR=2.279, 95% CI: 1.862-2.788), sleep disorders-osteoarticular class( OR=1.370, 95% CI: 1.111-1.691), and cardiovascular-metabolic class( OR=1.185, 95% CI: 1.003-1.400)were all found to be significantly associated with frailty. Conclusions:The cardiovascular-metabolic class is the most prevalent among elderly individuals.Various patterns exhibit distinct associations with frailty, with the complex multimorbidity class and the stroke-respiratory class being the most significant, as they markedly elevate the risk of frailty.