OBJECTIVE To investigate the apoptosis-inducing effect of esculetin （Esc） on acute myeloid leukemia （AML） HL-60 cells by regulating the protein kinase B （AKT）/S-phase kinase-associated protein 2 （SKP2）/MutT homolog 1 （MTH1） pathway. METHODS AML HL-60 cells were randomly divided into control group （routine culture）， Esc low-concentration group （L-Esc group， 25 μmol/L Esc）， Esc medium-concentration group （M-Esc group， 50 μmol/L Esc）， Esc high-concentration group （H-Esc group， 100 μmol/L Esc）， and high-concentration of Esc+ SC79 （AKT agonist） group （100 μmol/L Esc+5 μmol/L SC79）. Cell proliferation in each group was detected by MTT assay and colony formation assay. The level of reactive oxygen species （ROS） in cells was measured by using the CM-H2DCFDA fluorescent probe. Cell apoptosis was analyzed by flow cytometry. Western blot assay was performed to detect the expression levels of apoptosis-related proteins [B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cleaved caspase-3]， AKT/SKP2/MTH1 pathway-related proteins （p-AKT， AKT， SKP2， MTH1）， along with the upstream and downstream proteins of AKT phosphatidylinositol 3-kinase （PI3K）， cyclin-dependent kinase inhibitor 1 （P21） and cyclin-dependent kinase inhibitor 1B （P27）. RESULTS Compared with control group， the cell viability， colony number， and the phosphorylation levels of AKT and PI3K proteins as well as protein expressions of SKP2， MTH1 and Bcl-2 were significantly decreased （P＜0.05）， while ROS level， apoptosis rate， and the expression levels of Bax， cleaved caspase-3， P21 and P27 proteins were significantly increased （P＜0.05）. Moreover， the effects of Esc exhibited concentration-dependence （P＜0.05）. Compared with H-Esc group， above indexes of high-concentration of Esc+ SC79 group were reversed significantly （P＜0.05）. CONCLUSIONS Esc may promote massive ROS production and induce activation of apoptosis in HL-60 cells by inhibiting the AKT/SKP2/MTH1 pathway， thus inhibiting the proliferation of HL-60 cells.

Antibody-drug conjugates （ADCs） are a novel class of anti-tumor agents composed of a targeted monoclonal antibody， a cytotoxic drug， and a linker connecting the two. They combine the high specificity of antibodies with the potent cytotoxicity of chemotherapeutic agents. Triple-negative breast cancer （TNBC） is characterized by high aggressiveness， elevated risks of recurrence and metastasis， and poor prognosis， largely due to the lack of effective therapeutic targets. This review summarizes the research progress of ADCs in the treatment of TNBC. It has been found that ADCs targeting human epidermal growth factor receptor 2 （such as trastuzumab deruxtecan）， trophoblast cell surface antigen 2 （such as sacituzumab govitecan and datopotamab deruxtecan）， zinc transporter LIV-1 （such as ladiratuzumab vedotin）， HER-3 （such as patritumab deruxtecan）， epidermal growth factor receptor （such as AVID100）， and glycoprotein non-metastatic melanoma protein B （such as glembatumumab vedotin） have all demonstrated promising therapeutic effects against TNBC. Despite challenges including acquired resistance and treatment-related toxicities， ADCs are undoubtedly reshaping the therapeutic landscape for TNBC and are expected to occupy a more central position in TNBC treatment in the future.

Oral squamous cell carcinoma (OSCC) is a common head and neck malignancy. Approximately 50% to 60% of patients with OSCC are diagnosed at a locally advanced stage (clinical staging III-IVa). Even with comprehensive and sequential treatment primarily based on surgery, the 5-year overall survival rate remains below 50%, and patients often suffer from postoperative functional impairments such as difficulties with speaking and swallowing. Programmed death receptor-1 (PD-1) inhibitors are increasingly used in the neoadjuvant treatment of locally advanced OSCC and have shown encouraging efficacy. However, clinical practice still faces key challenges, including the definition of indications, optimization of combination regimens, and standards for efficacy evaluation. Based on the latest research advances worldwide and the clinical experience of the expert group, this expert consensus systematically evaluates the application of PD-1 inhibitors in the neoadjuvant treatment of locally advanced OSCC, covering combination strategies, treatment cycles and surgical timing, efficacy assessment, use of biomarkers, management of special populations and immune related adverse events, principles for immunotherapy rechallenge, and function preservation strategies. After multiple rounds of panel discussion and through anonymous voting using the Delphi method, the following consensus statements have been formulated: 1) Neoadjuvant therapy with PD-1 inhibitors can be used preoperatively in patients with locally advanced OSCC. The preferred regimen is a PD-1 inhibitor combined with platinum based chemotherapy, administered for 2-3 cycles. 2) During the efficacy evaluation of neoadjuvant therapy, radiographic assessment should follow the dual criteria of Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1 and immune RECIST (iRECIST). After surgery, systematic pathological evaluation of both the primary lesion and regional lymph nodes is required. For combination chemotherapy regimens, PD-L1 expression and combined positive score need not be used as mandatory inclusion or exclusion criteria. 3) For special populations such as the elderly (≥ 70 years), individuals with stable HIV viral load, and carriers of chronic HBV/HCV, PD-1 inhibitors may be used cautiously under the guidance of a multidisciplinary team (MDT), with close monitoring for adverse events. 4) For patients with a poor response to neoadjuvant therapy, continuation of the original treatment regimen is not recommended; the subsequent treatment plan should be adjusted promptly after MDT assessment. Organ transplant recipients and patients with active autoimmune diseases are not recommended to receive neoadjuvant PD-1 inhibitor therapy due to the high risk of immune related activation. Rechallenge is generally not advised for patients who have experienced high risk immune related adverse events such as immune mediated myocarditis, neurotoxicity, or pneumonitis. 5) For patients with a good pathological response, individualized de escalation surgery and function preservation strategies can be explored. This consensus aims to promote the standardized, safe, and precise application of neoadjuvant PD-1 inhibitor strategies in the management of locally advanced OSCC patients.

Objective: To study the association between ankyloglossia and sagittal mandibular development impairment as well as lower anterior dental crowding, providing a reference for clinical practice. Methods: This study was approved by the hospital's Medical Ethics Committee. A total of 100 patients aged 7-13 years were enrolled from January 2024 to January 2025, comprising 50 patients with ankyloglossia (case group) and 50 individuals with a healthy lingual frenulum (normal group). Clinical examination was performed to assess lingual frenulum length, Kotlow classification, and the simplified Hazelbaker assessment tool for lingual frenulum function (HATLFF) score. Cephalometric radiographs were used to measure the A-point-nasion-B-point (ANB) angle, sella-nasion-B-point (SNB) angle, and mandibular total length (condylion-gnathion [Co-Gn]). Dental cast analysis was conducted to evaluate lower anterior teeth crowding. Data were compared between the two groups. Pearson correlation analysis was used to examine the relationships between the lingual frenulum length, simplified HATLFF score, and cephalometric/dental cast parameters (ANB, SNB, Co-Gn, lower anterior crowding). The diagnostic value of ankyloglossia for mandibular development and lower anterior crowding was analyzed using receiver operating characteristic (ROC) curves. Results: Ankyloglossia was significantly associated with mandibular development and lower anterior crowding (P<0.05). The case group showed significantly lower values for the lingual frenulum length, simplified HATLFF score, SNB angle, and Co-Gn, while the ANB angle and lower anterior crowding index were significantly higher compared to the normal group (P<0.05). The lingual frenulum length and simplified HATLFF score were negatively correlated with the ANB angle and lower anterior crowding index, and positively correlated with the SNB angle and Co-Gn (P<0.05). ROC curve analysis indicated that the area under the curve (AUC) for the simplified HATLFF score, and ankyloglossia in predicting mandibular development deficiency and lower anterior crowding was greater than 0.700, demonstrating good diagnostic value. Conclusion A significant correlation exists between ankyloglossia and both mandibular development deficiency and lower anterior crowding.

Objective: To explore the latent profiles of psychological help seeking intentions among adolescents in Wuhan, and to investigate their association with non suicidal self injury (NSSI), in order to provide a theoretical basis for constructing an early intervention system for adolescents NSSI. Methods: From October to December 2022, a convenient sampling method was used to select 3 975 students from grades 7 to 12 in Wuhan. A self administered questionnaire assessed psychological help seeking intentions, followed by a post survey interview using the Ottawa Self injury Inventory to evaluate NSSI behaviors. Latent profile analysis(LPA) was used to explore the potential categories of help seeking intentions in adolescents. Multinomial Logistic regression was used to analyze factors associated with the latent profiles of help seeking intentions and multiple Logistic regression was used to analyze the association between latent profiles of help seeking intentions and NSSI. Results: Based on latent profile analysis, four latent profiles of help seeking intentions in adolescents were identified, namely overall low, moderate, and high help seeking group, as well as family/friendfocused help seeking group, accouting for 14.1%, 20.9%, 43.1% and 21.8%, respectively. Multinomial Logistic regression analysis showed that compared to senior high school students, junior high school students were more willing to seek help from family and friends ( OR =1.56); compared to males, females were more likely to exhibit moderate help seeking intentions( OR =1.37); students with extroverted or balanced personalities were more likely to exhibit high level help seeking intentions( OR =1.50, 1.49); students with average or good family economic status, better parental relationships, and adequate mental health knowledge were more likely to exhibit moderate and high level help seeking intentions( OR =1.59, 2.02; 1.80, 2.64; 1.44, 1.55; 1.34, 1.58) (all P <0.05). Students with moderate or severe family dysfunction were less likely to seek help from family and friends or to exhibit moderate and high level help seeking intentions( OR =0.51, 0.60, 0.25; 0.22, 0.27, 0.06, all P <0.01). Students whose parents exhibited stigma towards mental illness were less likely to show high level help seeking intentions( OR= 0.78 , P <0.05). Multivariable Logistic regression analysis results indicated that compared to the overall low help seeking intentions group, reduced risk of NSSI was observed among students in the overall moderate and overall high groups, as well as in the family/friend focused help seeking group( OR =0.73, 0.60, 0.70, all P <0.05). Conclusions Active psychological help seeking intentions can reduce NSSI behaviors in adolescents. Interventions should focus on improving family support environment to enhance adolescents intentions to seek psychological help.

ObjectiveTaking classical herbal pair compatibility research as the entry point， this study aimed to deeply investigate the material basis and compatibility rules underlying the antidepressant effects of the traditional Chinese medicine （TCM） formula Zhizi Houpotang， and to elucidate its antidepressant mechanism， with a particular focus on its regulation of neuroinflammatory responses mediated by the NOD-like receptor protein 3 （NLRP3）/gasdermin D （GSDMD） signaling pathway and the consequent improvement of neuronal synaptic plasticity. MethodsC57BL/6J mice were randomly divided into a blank control group， a chronic unpredictable mild stress （CUMS） depression model group， a Zhizi Houpotang full-formula group （6 g·kg^-1·d^-1）， a Magnoliae Officinalis Cortex （MOC）-Aurantii Fructus Immaturus （AFI） herbal pair group （4.2 g·kg^-1·d^-1）， a Gardeniae Fructus （GF）-MOC herbal pair group （4.2 g·kg^-1·d^-1）， a GF-AFI herbal pair group （3.6 g·kg^-1·d^-1）， and a positive drug group （fluoxetine， 12 mg·kg^-1·d^-1）. Depressive-like behaviors in mice were evaluated using behavioral tests. Immunofluorescence staining was used to label and quantify the expression of the microglial marker ionized calcium-binding adaptor molecule 1 （Ibal） and the purinergic receptor P2X ligand-gated ion channel 7 （P2RX7） in the prefrontal cortex （PFC）. Enzyme-linked immunosorbent assay （ELISA） was applied to detect the levels of inflammatory cytokines interleukin-1β （IL-1β） and interleukin-18 （IL-18） in serum and PFC tissues. Western blot was employed to determine the expression of pannexin 1 （Panx1）， P2RX7， NLRP3， apoptosis-associated speck-like protein containing a CARD （ASC）， Caspase-1， GSDMD， postsynaptic density protein 95 （PSD95）， and the presynaptic protein Synapsin 1 in PFC tissues. Golgi staining was used to assess dendritic spine density of neurons in the PFC. ResultsCompared with the blank control group， the depression model group exhibited significant depressive-like behaviors. In addition， the immunofluorescence areas of Ibal and P2RX7 in the PFC were significantly increased （P<0.01）， the levels of IL-1β and IL-18 in serum and the PFC were significantly elevated （P<0.01）， and the protein expression levels of Panx1， P2RX7， NLRP3， ASC， Caspase-1， and GSDMD in the PFC were significantly upregulated （P<0.01）. In contrast， the protein expression levels of PSD95 and Synapsin 1 were significantly downregulated （P<0.01）， and neuronal dendritic spine density was significantly reduced （P<0.01）. Compared with the model group， the Zhizi Houpotang full-formula group and the GF-MOC herbal pair group showed significant improvement in all the above indicators （P<0.01）. The GF-AFI herbal pair group improved all the above indicators except P2RX7， Caspase-1， GSDMD， and PSD95 （P<0.05， P<0.01）. In contrast， the MOC-AFI herbal pair group showed no statistically significant improvement in any of the above indicators compared with the model group. ConclusionZhizi Houpotang and its key herbal pair， GF-MOC， can effectively ameliorate CUMS-induced depressive-like behaviors in mice. Its core antidepressant mechanism may involve inhibition of P2RX7/Panx1 signaling， thereby blocking the NLRP3/GSDMD-mediated pyroptosis pathway and significantly reducing the release of inflammatory cytokines IL-1β and IL-18. Simultaneously， it upregulates the expression of synapse-related proteins PSD95 and Synapsin 1 and increases dendritic spine density， promoting the recovery of synaptic plasticity. These results suggest that GF plays a key role in the antidepressant effects of this formula， and that the compatibility of GF with MOC may represent the principal herbal pair combination responsible for its core therapeutic action.

ObjectiveTo identify the independent risk factors for pyloric lymph node （PLN） metastasis in patients with upper gastric cancer （UGC） and to construct a nomogram prediction model applicable for UGC patients. MethodsClinical data of 823 UGC patients attended between January 2020 and November 2023 were retrospectively collected. Patients were randomly divided into a training set （n=576） and a validation set （n=247） at a 7∶3 ratio. Based on the training set， multivariate Logistic regression analysis was performed to identify independent risk factors for PLN metastasis， and a nomogram prediction model was constructed accordingly. The model's discriminative ability and calibration were assessed using receiver operating characteristic （ROC） curves and calibration curves. Finally， external validation was conducted using the validation set to evaluate the model's stability and generalizability. ResultsMultivariate Logistic regression analysis revealed that tumor size （OR=1.324， 95%CI： 1.053-1.667）， T3 stage （OR=5.738， 95%CI： 1.281-25.695）， T4 stage （OR=7.680， 95%CI： 1.542-38.247）， lymphovascular invasion （LVI） （OR=6.623， 95%CI： 1.384-31.708）， differentiation extent （OR=3.108， 95%CI： 1.545-6.251）， and fibrinogen degradation product （FDP） level （OR=4.849， 95%CI： 2.071-11.355） were independent risk factors for PLN metastasis in UGC patients.The nomogram model constructed based on these factors demonstrated areas under the ROC curve （AUC） of 0.815 （95%CI： 0.751-0.815） in the training set and 0.832 （95%CI： 0.731-0.933） in the validation set. Calibration curves indicated good agreement between predicted and observed outcomes. ConclusionThis nomogram prediction model exhibits good predictive performance for assessing the risk of PLN metastasis in UGC patients.

Objective To investigate the current distribution and predict the future suitable habitats of Haemaphysalis longicornis in Nanjing City, so as to provide insights into control and early warning of ticks and management of tick-borne diseases in Nanjing City. Methods The electronic map of Nanjing City was obtained from the National Platform for Common GeoSpatial Information Services. The distribution of H. longicornis and the longitude and latitude of distribution points from 2022 to 2024 were obtained from centers for disease control and prevention across each district in Nanjing City. Climatic and environmental variable data in Nanjing City were captured from the Worldclim database. Initially, 19 bioclimatic variables in this database were selected, including annual mean temperature, mean diurnal range, isothermality, temperature seasonality, maximum temperature of the warmest month, minimum temperature of the warmest month, temperature annual range, mean temperature of the wettest quarter, mean temperature of the driest quarter, mean temperature of the warmest quarter, mean temperature of the coldest quarter, annual precipitation, precipitation of the wettest month, precipitation of the driest month, precipitation seasonality, precipitation of the wettest quarter, precipitation of the driest quarter, precipitation of the warmest quarter, and precipitation of the coldest quarter. The elevation and normalized difference vegetation index were obtained from Data Sharing Platform of the Center for Resources and Environmental Sciences, Chinese Academy of Sciences. Then, the distribution points of H. longicornis, elevation, vegetation index and 19 bioclimatic variables were loaded into the software MaxEnt 3.4.4 to evaluate and screen out the variables with a contribution rate of 1% and higher. ArcGIS 10.8.1 software was used to extract the elevation, vegetation index and 19 bioclimatic variables of the distribution points of H. longicornis for a correlation analysis. If the absolute value of the correlation coefficient was 0.8 and higher, the variable with the higher contribution was retained. The 2050 dataset of the BCCCSM2-MR atmospheric circulation model in the coupled model intercomparison project phase 6 (CMIP6) were obtained from the Worldclim database as climate data for 2050. Screened H. longicornis species data and environmental and climate data were loaded into the maximum entropy (MaxEnt) model with the software MaxEnt 3.4.4 for training and validation, and then, all data generated from the model were imported into the software ArcGIS 10.8.1 to generate raster data and yield the map pertaining to the distribution of H. longicornis risk in Nanjing City. The accuracy of the model was evaluated with a receiver operating characteristic (ROC) curve, and the predictive effect of the model was assessed with area under the ROC curve (AUC). The suitable habitats of H. longicornis were classified in Nanjing City with the software ArcGIS 10.8.1, and the areas of distribution of suitable habitats in various categories were recorded to create the map of current H. longicornis suitable habitats classification in Nanjing City. The climatic and geographic information data in 2050 were employed as future environmental and climatic factors, and current environmental and climatic factors and current H. longicornis distribution data were additionally used to predict the future suitable habitats of H. longicornis in Nanjing City. In addition, the contributions of environmental and climatic factors to distribution of suitable habitats of H. longicornis was evaluated with the Jackknife method in Nanjing City. Results A total of 10 environmental and climatic variables were screened for analysis of the suitability of H. longicornis in Nanjing City based on correlation analyses and contributions of the MaxEnt model, including annual mean temperature, precipitation of the warmest quarter, vegetation index, precipitation of the wettest month, temperature annual range, annual precipitation, mean temperature of the warmest quarter, elevation, mean temperature of the wettest quarter, and maximum temperature of the warmest month, and annual mean temperature (34.8%), precipitation of the warmest quarter (17.3%), vegetation index (13.1%), and precipitation of the wettest month (10.8%) contributed relatively highly to the distribution of suitable habitats of H. longicornis in Nanjing City. The mean AUC of the ROC curve was 0.810 ± 0.055 for 10 repeated modeling results of the MaxEnt model, indicating high predictive performance of the model. The potential distribution areas of H. longicornis were predicted to be mainly located in Luhe District, Pukou District, Jiangning District, Lishui District, and Gaochun District in Nanjing City with the MaxEnt model. Under current climatic conditions, the area of potential suitable habitats of H. longicornis was 4 182.42 km² in Nanjing City, including 1 252.94 km² highly suitable habitats, which accounted for 19.00% of the total area of Nanjing City. Under the climate scenario in 2050, the area of potential suitable habitats of H. longicornis was projected to increase to 5 467.58 km² in Nanjing City, accounting for 82.95% of the total area of the city, and these habitats were mainly concentrated in Luhe District, Pukou District, Jiangning District, Lishui District, and Gaochun District. The areas of suitable habitats of H. longicornis at various categories were predicted to vary greatly in 2050, and the area of highly suitable habitats of H. longicornis was projected to increase to 2 378.82 km², accounting for 36.08% of the total area of Nanjing City. Based on jackknife tests and contributions of environmental and climatic variables, 6 dominant environmental and climatic factors were screened, including annual mean temperature (34.8% contribution), precipitation of the warmest quarter (17.3% contribution), vegetation index (13.1% contribution), precipitation of the wettest month (10.8% contribution), temperature annual range (5.4% contribution), and mean temperature of the warmest quarter (5.0% contribution), with cumulative contributions of 86.4%. Conclusion The distribution of H. longicornis is strongly associated with vegetation, temperature and precipitation in Nanjing City. Future climate change may lead to an expansion of the distribution area of H. longicornis in Nanjing City.

Background Pyrethroid pesticides (PYRs) can cross the placental barrier to cause intrauterine fetal exposure, which may lead to developmental immunotoxicity (DIT). However, the specific effect of maternal PYR exposure during pregnancy on the cellular immune function of 1-year-old children remains unclear. Objective To explore the effect of PYRs exposure throughout the entire pregnancy on peripheral blood lymphocytes in 1-year-old children and potential sensitive window period of PYRs exposure. Methods A birth cohort was established by enrolling pregnant women in their first trimester and following them and their infants until one year of age. Ultra-high performance liquid chromatography-tandem mass spectrometry was used to detect the levels of PYRs metabolites, including 3-phenoxybenzoic acid (3PBA), 4-fluoro-3-phenoxybenzoic acid (4F3PBA), and cis-3-(2,2-dichlorovinyl)-2,2- dimethylcyclopropane carboxylic acid (cis-DBCA), in the urine of pregnant women during the first trimester (gestational weeks 6-12), the second trimester (gestational weeks 21-24), and the third trimester (gestational weeks 33-36). Peripheral blood leukocyte and lymphocyte counts were measured in children at 12 months of age using the Coulter principle combined with flow cytometry. Exposure levels of PYRs metabolites in each trimester were divided into low, moderate, and high exposure groups based on the 25th (P₂₅) and 75th (P₇₅) percentiles. Meanwhile, participants were classified as having repeated high or low exposure if their metabolite levels were > P₇₅ or <P₂₅ in at least two trimesters, respectively, while all others were categorized as having repeated moderate exposure. Generalized linear models were used to analyze the associations between trimester-specific and repeated PYRs metabolite exposure levels and the peripheral blood white blood cell (WBC) and lymphocyte counts in children aged 1 year. Results A total of 336 mother-child pairs were included in this study. For the pregnant women, the total detection rates of maternal urinary 3PBA, 4F3PBA, and cis-DBCA across the three trimesters of pregnancy were 80.5%, 100.0%, and 81.3%, respectively; and median creatinine-corrected concentrations were 0.24, 0.36, and 0.42 μg·g⁻¹, respectively. In children aged 1 year, the mean WBC and lymphocyte counts in peripheral blood were (8.9±2.0)×10⁹·L⁻¹ and (5.7±1.6)×10⁹·L⁻¹, respectively. The results of the generalized linear model analysis indicated that compared to the low exposure group, the high cis-DBCA exposure group during the third trimester of pregnancy had significantly lower peripheral blood WBC count (β=−0.87, 95%CI: −1.51, −0.23) and lymphocyte count (β=−0.64, 95%CI: −1.15, −0.13); and the repeated high-exposure group of cis-DBCA had significantly lower peripheral blood WBC count (β=−1.34, 95%CI: −2.34, −0.34) and lymphocyte count (β=−0.80, 95%CI: −1.60, −0.01) than the repeated low exposure group. Similarly, the repeated moderate-exposure group of cis-DBCA had a significantly lower peripheral blood WBC count (β=−0.83, 95%CI: −1.59, −0.07) than the repeated low exposure group. Conclusion High maternal exposure to PYRs with cis-DBCA as the major metabolite exposure is associated with decreased peripheral leukocyte and lymphocyte counts in children aged 1 year, and repeated high-level exposure throughout gestation appears to exacerbate DIT in offspring. The third trimester of pregnancy maybe a sensitive window for children's DIT induced by exposure to PYRs during pregnancy.

BACKGROUND:Nanocell vesicles possess functions such as re-epithelialization,antioxidation,anti-inflammation,and regulation of extracellular matrix remodeling.Meanwhile,apoptotic bodies have the immunomodulatory effects.Therefore,the combination of the two to form nanofusion vesicles can synergistically promote the healing of diabetic skin wounds.OBJECTIVE:To elucidate the impact of nanofusion vesicles on skin wound healing in a diabetic murine model.METHODS:(1)Material preparation and characterization:The primary bone marrow mesenchymal stem cells of C57BL/6J neonatal mice and the neutrophil apoptotic bodies of C57BL/6J mice were isolated and extracted.The nanofusion vesicles were prepared by micro-extrusion mechanism.(2)In vitro experiment:MTT assay was used to detect the proliferative effect of different concentrations of nanofusion vesicles on NIH-3T3 cells and human umbilical vein endothelial cells.Reactive oxygen species fluorescence probe was used to detect the antioxidant effect of nano-fusion vesicles on NIH-3T3 cells treated with hydrogen peroxide(H2O2).The inhibitory effect of nanofusion vesicles on RAW 264.7 macrophage inflammation induced by lipopolyside was detected by real-time quantitative RT-qPCR.(3)In vivo experiment:36 male C57BL/6J mice were employed to develop a murine model of diabetes mellitus.Following the successful induction of diabetes,two circular full-thickness wounds,each with a diameter of 6 mm,were created on either side of the diabetic mice's spine using a skin punch.The mice were divided into three groups by random number table method.The control group was injected with 0.1 mL of phosphate buffer solution.The nanovesicle group was injected with 0.1 mL nanovesicles(25 μg/mL).The nanofusion vesicle group was injected with 0.1 mL nanofusion(25 μg/mL)vesicles.After treatment for three consecutive days,the wound healing and histomorphological changes were observed.RESULTS AND CONCLUSION:(1)In vitro experiment:nanofusion vesicles,when administered at concentrations ranging from 0 to 100 μg/mL,exhibited no toxic effects and promoted the proliferation of NIH-3T3 and HUVEC cell lines.Notably,a concentration of 25 μg/mL nanofusion vesicle significantly enhanced the proliferation of NIH-3T3 cells.Furthermore,the survival rate of human umbilical vein endothelial cells was observed to increase in correlation with escalating concentrations of nanofusion vesicles.Nanofusion vesicles had a good antioxidant effect.In comparison to the H2O2 group,the fluorescence signal indicative of reactive oxygen species was progressively diminished in both the nanovesicle group and the nanofusion vesicle group.Furthermore,nanofusion vesicles possessed anti-inflammatory capabilities,effectively mitigating the inflammatory response in macrophages triggered by lipopolysaccharide stimulation.(2)In vivo experiment:Hematoxylin-eosin and Masson's trichrome staining revealed that in comparison to the control group,both the nanovesicle group and the nanofusion vesicle group exhibited a significant increase in granulation tissue formation and collagen fiber deposition within the wounds by day 6.Notably,the nanofusion vesicle group displayed the most pronounced effects.On day 12,the wound of nanofusion vesicle group was significantly reduced,and the healing rate was significantly faster than that of other groups(P＜0.01),and the effect of promoting wound healing was the most significant.Our findings demonstrated that nanofusion vesicles exhibited superior pro-cell proliferative,antioxidant,and anti-inflammatory properties,thereby exerting a beneficial effect on the promotion of skin wound healing in diabetic mouse models.