Objective To investigate the protective effect and underlying mechanism of human umbilical cord mesenchymal stem cell-derived exosomes (hucMSC-Exo) on renal ischemia-reperfusion injury (IRI). Methods hucMSC-Exos were isolated and characterized. A mouse renal IRI model was established and the animals were divided into Sham, IRI, IRI+hucMSC-Exo, IRI+hucMSC-Exo+JY-2 and Sham+JY-2 groups. Serum creatinine (Scr) and blood urea nitrogen (BUN) were measured. Hematoxylin-eosin (HE) staining was used to evaluate renal histopathology. Enzyme-linked immune absorbent assay was performed to determine serum interleukin (IL)-1β and IL-18 levels. Western blotting was used to detect the expression of activating transcription factor 3 (ATF3), Toll-like receptor 4 (TLR4), nuclear factor (NF)-κB, NOD-like receptor protein 3 (NLRP3), cysteineyl aspartate specific proteinase (Caspase)-1 p20 and Gasdermin D(GSDMD). Real-time fluorescent quantitative polymerase chain reaction was employed to measure ATF3, TLR4 and NF-κB messenger RNA (mRNA). Immunohistochemistry was conducted to examine NLRP3, Caspase-1 p20 and GSDMD. An in vitro hypoxia/reoxygenation (H/R) model was established in HK-2 cells and divided into Control, H/R, H/R+hucMSC-Exo, H/R+hucMSC-Exo+JY-2 and Control+JY-2 groups. Western blotting was used to detect the expression of ATF3, TLR4 and NF-κB. Real-time fluorescent quantitative polymerase chain reaction was used to measure NLRP3, GSDMD and Caspase-1 mRNA. Results HucMSC-Exos were successfully isolated and identified. Compared with the Sham group, the IRI group exhibited elevated Scr and BUN, higher tubular injury scores, increased protein expression levels of ATF3, TLR4, NF-κB p65, NLRP3, Caspase-1 p20 and GSDMD, and raised mRNA expression levels of ATF3, TLR4, NF-κB. Compared with the IRI group, the IRI+hucMSC-Exo group showed decreased Scr and BUN, lower tubular injury scores, up-regulated ATF3 protein and mRNA, down-regulated TLR4, NF-κB p65, NLRP3, Caspase-1 p20 and GSDMD protein, and declined TLR4 and NF-κB mRNA. Compared with the IRI+hucMSC-Exo group, the IRI+hucMSC-Exo+JY-2 group exhibited increased Scr and BUN levels, elevated renal tubular injury scores, decreased ATF3 protein expression levels, elevated protein expression levels of TLR4, NF-κB p65, NLRP3, Caspase-1 p20, and GSDMD, decreased ATF3 mRNA expression levels, and elevated mRNA expression levels of TLR4 and NF-κB. (all P < 0.05). Compared with the Control group, the expression levels of ATF3, TLR4 and NF-κB p65 proteins were increased in the H/R group, and the expression levels of NLRP3, Caspase-1 and GSDMD mRNA were increased. Compared with the H/R group, the expression level of ATF3 protein was increased, the expression levels of TLR4 and NF-κB p65 proteins were decreased, and the expression levels of NLRP3, Caspase-1 and GSDMD mRNA were decreased in the H/R+hucMSC-Exo group. Compared with the H/R+hucMSC-Exo group, the expression level of ATF3 protein was decreased, the expression levels of TLR4 and NF-κB p65 proteins were increased, and the expression levels of NLRP3, Caspase-1 and GSDMD mRNA were increased in the H/R+hucMSC-Exo+JY-2 group (all P < 0.05). Conclusions HucMSC-Exos alleviate renal IRI by up-regulating ATF3, thereby negatively regulating the TLR4/NF-κB signaling pathway and subsequently inhibiting pyroptosis.

BackgroundThe middle school stage represents a crucial period for the development of borderline personality features. Negative parenting styles and emotional regulation strategies are associated with the formation of borderline personality features. However， the moderating role of emotional regulation strategies between negative parenting styles and borderline personality features among middle school students remains unclear. ObjectiveTo explore the moderating influence of emotional regulation strategies in the relationship between negative parenting styles and borderline personality features among middle school students， and to provide references for the intervention of borderline personality features. MethodsIn October 2023， a total of 5 965 middle school students from three middle schools in Nanning， Guangxi Zhuang Autonomous Region were selected by cluster sampling， and assessed by the Borderline Personality Features Scale for Children （BPFS-C）， the Egna Minnen Barndoms Uppfostran （EMBU）， and the Emotion Regulation Questionnaire-Chinese Revised Version （ERQ-CRV）. Pearson correlation analysis was used to test the correlation between the scores of each scale， and the model 1 of the Process macro program was used to conduct the moderating effect test. ResultsA total of 5 572 middle school students （93.41%） completed this study， and 1 388 of them （24.91%） were identified as having high borderline personality features. The BPFS-C score of middle school students was positively correlated with the score of the negative parenting style dimension of EMBU （r=0.367， P<0.01）， negatively correlated with the score of the cognitive reappraisal dimension of ERQ-CRV （r=-0.168， P<0.01）， and positively correlated with the score of the expression inhibition dimension of ERQ-CRV （r=0.344， P<0.01）. Cognitive reappraisal played a negative moderating effect between negative parenting styles and borderline personality features （β=-0.072， 95% CI： -0.104–-0.041， P<0.01）， while expressive suppression played a positive moderating effect （β=0.076， 95% CI： 0.055–0.097， P<0.01）. ConclusionCognitive reappraisal strategy may help mitigate the negative influence of negative parenting styles on middle school students' borderline personality features， while expressive suppression may exacerbate the harm of negative parenting styles to the borderline personality features of middle school students.

Objective: To evaluate the effects of green tea extract (GTE) and its main catechin monomers on erythrocyte lesion (such as hemolysis, decreased energy metabolism and oxidative stress) during in vitro storage, and to explore its potential as a novel additive for erythrocyte preservation solutions. Methods: The composition of GTE was analyzed by high-performance liquid chromatography (HPLC). Using an in vitro simulated storage model, erythrocytes were stored in CPDA-1 preservation solution supplemented with GTE and the three most abundant catechin monomers (EGCG, ECG, EGC) for 60 days. Hemolysis rate and ATP content were dynamically monitored during storage. Flow cytometry was used to analyze phosphatidylserine (PS) exposure. Meanwhile, the protective effects of each component were verified in an acute oxidative stress model, and erythrocyte membrane stability was assessed by osmotic fragility test. Results: After 60 days of storage at 4℃, the hemolysis rate at the end of storage in the GTE group was <0.8%, which was superior to that in the control group and the single catechin-supplemented groups. Erythrocyte osmotic fragility assay showed that GTE could enhance the stability of erythrocyte membranes. In the acute oxidative stress experiment, the protective rate of GTE against erythrocyte injury exceeded 99%, which was better than that of the single catechin groups. At the initial stage of storage, ATP content decreased in all catechin-treated groups, but PS exposure was not significantly increased. Conclusion: The addition of GTE can effectively alleviate storage lesions of erythrocytes, with efficacy superior to that of single catechins. GTE enhances the antioxidant capacity and membrane stability of stored erythrocytes. Our results provide new experimental evidence for the development of GTE-based erythrocyte preservation additives.

Objective:To explore the impact of adolescent mental health status on smartphone addiction, and construct a predictive model for smartphone addiction based on the eXtreme Gradient Boosting(XGBoost) algorithm and multivariate Logistic regression.Methods:In April 2023, a cross-sectional survey was conducted among 14 666 adolescents.All participants were systematically evaluated using a self-developed general information questionnaire, the middle school student mental health scale(MSSMHS), the adolescents self-harm scale(ASHS), the interaction anxiousness scale(IAS), the mobile phone addiction index(MPAI), the middle school students shame scale(MSSS), the UCLA loneliness scale(UCLA-LS), the multidimensional peer victimization scale(MPVS), and the basic psychological needs scale(BPNS).R software version 4.3.2 was used for data analysis. Participants were randomly divided into training set and validation set at the ratio of 7∶3.The XGBoost model and multivariate logistic regression model were constructed to predict the risk of smartphone addiction, and a nomogram was plotted.Model performance was evaluated using the Hosmer-Lemeshow test, area under the curve(AUC), and accuracy(ACC).Results:(1) A total of 14 036 high school students were included in the study, with 5 069(36.1%) exhibited smartphone addiction.The training set comprised 9 826 students, with 3 549(36.1%) being smartphone addicts.The validation set included 4 210 students, with 1 520(36.1%) being smartphone addicts.(2) The XGBoost model identified shame-proneness and social anxiety as the two main predictors of smartphone addiction.(3) Multivariate Logistic regression analysis revealed that anxiety( B=0.328, OR(95% CI)=1.39(1.07-1.81), P=0.015), interpersonal sensitivity( B=0.311, OR(95% CI)=1.36(1.05-1.77), P=0.018), learning pressure( B=0.606, OR(95% CI)=1.83(1.46-2.31), P<0.001), mood swings( B=0.775, OR(95% CI)=2.17(1.70-2.78), P<0.001), social anxiety( B=0.024, OR(95% CI)=1.02(1.01-1.04), P<0.001), shame-proneness( B=0.049, OR(95% CI)=1.05(1.04-1.06), P<0.001), and peer victimization( B=0.037, OR(95% CI)=1.04(1.02-1.06), P<0.001) were significant predictors of smartphone addiction.(4) The ACC and AUC values of the XGBoost model were 0.890 and 0.929 in the training set, and 0.865 and 0.864 in the validation set, respectively.The multivariate Logistic regression model achieved ACC and AUC values of 0.870 and 0.854 in the training set, and 0.867 and 0.859 in the validation set, respectively. Conclusion:Anxiety, interpersonal sensitivity, learning pressure, mood swings, social anxiety, shame-proneness, and peer victimization are identified risk predictors of smartphone addiction in high school adolescents.

Pyroptosis,a form of programmed cell death primarily mediated by gasdermin D(GSD-MD),plays a crucial role in infections and inflammatory responses.Although the OTU deubiquiti-nase 4(OTUD4)has shown its importance in various cellular processes,its specific function in py-roptosis remains unclear,which necessitates a deeper understanding of its role.This study aims to explore the role of OTUD4 in regulating GSDMD-mediated pyroptosis.Through experiments inclu-ding cell transfection,lactate dehydrogenase(LDH)release assays,co-immunoprecipitation(Co-IP),and Western blot(WB)analysis,we investigated the interaction between OTUD4 and GSD-MD and their influence on GSDMD-mediated pyroptosis.Our results indicated that OTUD4 could bind to GSDMD,significantly reduce Caspase-1-mediated cleavage of GSDMD,and lower protein levels of GSDMD-p30,thereby inhibiting pyroptosis.Conversely,the enzymatically inactive mutant of OTUD4(C45A)exhibited a markedly diminished inhibitory effect on pyroptosis.Furthermore,OTUD4 also demonstrated a significant inhibitory effect on pyroptosis in porcine cells.In conclu-sion,this study reveals that OTUD4 can inhibit GSDMD-p30-mediated pyroptosis through its deu-biquitinating activity,highlighting the critical function of OTUD4 in the regulation of pyroptosis and suggesting a potential target for the development of new therapeutic strategies.

Polygonatum sibiricum polysaccharide(PSP)is a polysaccharide with multiple pharma-cological activities that has been widely studied and used in the human body.However,there is cur-rently a lack of research investigating the potential advantages of PSP in poultry farming.This study investigated the effects of adding PSP to drinking water on the growth performance,antioxi-dant status,serum biochemical indicators,ileal tissue morphology,immune organs,and intestinal function of chicks.88 Hailan brown laying hens were randomly divided into 4 groups,with 22 hens in each group,namely the blank control group(CON),and fed with basic feed;The low-dose PSP group(250 mg/L),the medium dose PSP group(500 mg/L),and the high-dose PSP group(1 000 mg/L)were fed with corresponding doses of PSP through drinking water on the basis of basic feed,and the experimental period was 21 d.The initial and final body weight and immune or-gan relative quality of chicks,serum biochemical indicators,the activities of SOD,CAT,and GSH-Px,as well as the contents of T-AOC and MDA in the serum of chicks were measured;HE stai-ning method was used to observe the pathological changes of intestinal tissue slices in the ileum;Real time fluorescence quantitative PCR technology was used to detect the mRNA expression lev-els of cytokines ZO-1,Claudin-1,Occludin,Mucin-2,IL-1β,TNF-a,IFN-γ,IL-4,IL-8,and IL-10 in the ileum.The results showed that compared with the blank control group,the addition of medium dose PSP significantly increased the final relative quality(P＜0.01),the final body weight and ADG of PSP500 group chicks significantly increased(P＜0.01),and the F/G of PSP250 and PSP500 groups significantly decreased(P＜0.05 or P＜0.01).The villus height of the jejunum in the 200,500,and 1 000 mg/L PSP groups of chicks significantly increased(P＜0.05).The SOD ac-tivity significantly increased(P＜0.01),and the CAT activity in the PSP1000 group significantly increased(P＜0.01).The PSP500 and PSP1000 groups significantly reduced the mRNA expression of cytokines IL-1β,IL-4,and IFN-γ in the ileum(P＞0.05);PSP did not show significant changes in serum total protein(TP),albumin(ALB),glucose(GLU),cholesterol(T-CHO)content,and immune organ index(P＜0.05).In summary,PSP can improve growth performance,enhance an-tioxidant capacity,improve ileal morphology and epithelial barrier function,and regulate mucosal immune status.Considering the overall economic benefits,the recommended level of PSP addition is 500 mg/L.

AIM:To determine if scutellarin(Scu)provides neuroprotection by reducing neuronal apoptosis in rats subjected to middle cerebral artery occlusion(MCAO)via the inhibition of the JAK2/STAT3 signalling pathway.METHODS:Proteins linked to Scu and ischaemic stroke-induced neuronal apoptosis were identified using the Swiss Tar-get Prediction,PharmMapper,OMIM,and GeneCards databases.Intersecting targets were identified through Venn analy-sis.Protein-protein interaction networks were visualised utilising Cytoscape software,and principal targets were identi-fied.Enrichment analyses of GO functions and KEGG pathways were conducted utilising the Metascape database.Molecu-lar docking of Scu with core targets was performed utilising AutoDock Vina.The neuroprotective effects of Scu were as-sessed in MCAO rats using Zea Longa scoring and the suspension test.JAK2/STAT3 phosphorylation levels and apoptosis-related proteins[cleaved caspase-3(C-caspase-3),caspase-3,Bax,and Bcl-2]were assessed using Western blot and im-munofluorescence staining.The JAK2-specific inhibitor AG490 was employed to further investigate the role of the JAK2/STAT3 signaling pathway.RESULTS:Network pharmacology analysis revealed 832 shared targets,with pathways en-riched in tumor-associated pathways,the JAK/STAT signalling pathway,and the HIF-1 signalling pathway.Molecular docking revealed robust binding affinities of Scu with the ten principal targets.Behavioural assessments utilising Zea Lon-ga scoring and the suspension test demonstrated that Scu markedly enhanced neurological recovery in MCAO rats.Western blot and immunofluorescence analyses demonstrated that phosphorylation levels of JAK2 and STAT3,along with the ex-pression of C-caspase-3,Bax,and Bcl-2,were markedly elevated in the MCAO group relative to the sham group(P<0.05).Post Scu treatment,phosphorylation levels of JAK2 and STAT3,along with C-caspase-3 and Bax expression,were markedly diminished,whereas Bcl-2 expression and fluorescence intensity were substantially increased(P<0.05).In the combined AG490 and Scu treatment group(MCAO+Scu+AG490),the phosphorylation levels of JAK2 and STAT3,as well as the expression of C-caspase-3 and Bax,exhibited no significant difference when compared to the Scu-alone group(P>0.05).Bcl-2 expression and fluorescence intensity were markedly reduced in the combined AG490 and Scu treatment group relative to the Scu-alone group(P<0.05).CONCLUSION:Scu seems to provide neuroprotection in ischaemic stroke by reducing neuronal apoptosis through the inhibition of the JAK2/STAT3 signalling pathway.

Objective:To investigate the role of the Smad3 signaling pathway in the process of silica-induced epithelial-mesenchymal transition (EMT) .Methods:In September 2022, lung epithelial cells (BEAS-2B) were exposed to different concentrations of silica suspension (0, 50, 100, and 150 μg/ml) for 6 and 12 hours. Additionally, SIS3, a specific inhibitor of phosphorylated Smad3 (p-Smad3) , was utilized to establish the p-Smad3 inhibition model. The cells were divided into four groups: blank control gruop, silica group, SIS3 intervention group, and SIS3 +silica group. Cell morphology was observed using an inverted fluorescence microscope, while cell viability was assessed using a Cell Counting Kit-8 (CCK-8) . The mRNA and protein expression levels of E-cadherin (E-Cad) , N-cadherin (N-Cad) , Vimentin, Smad3, and p-Smad3 were analyzed by Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting, respectively. Differences between two groups were compared using Student's t-test, and multiple group comparisons were analyzed using a one-way analysis of variance with the Student-Newman-Keuls test.Results:Compared with the blank control group, the morphology of BEAS-2B cells shifted from epithelial to mesenchymal cell-like following silica exposure, and the cell viability of BEAS-2B cells declined after exposure to 150 μg/ml silica for 6 and 12 hours. Furthermore, silica exposure led to significant reductions in mRNA and protein expression levels of the epithelial cellular marker (E-Cad) in BEAS-2B cells, accompanied by increased expressions of interstitial cellular markers (N-Cad and Vimentin) . Importantly, the level of p-Smad3/Smad3 expression levels was also elevated in silica-treated cells ( P<0.05) . Compared to the blank control group, the level of p-Smad3/Smad3 expression levels was significantty reduced. Moreover, compared to the silica group, the protein expression levels of N-Cad and Vimentin in the cell of the SIS3+silica group were significantly reduced, while the E-Cad expression was increased ( P<0.05) . Conclusion:Silica exposure can prmote the epithelial mesenchymaol transformotion process by activating smod3 signa ling pathuay, and in hibiting smad3 signa ling pathuay can effctively alleviate the occurrence of epithelial mesenchymal transformation process.

Objective To develop an effective intervention strategy to improve medication adherence among chronic disease patients.Methods Based on the behavior change wheel theory and integrating literature analysis,questionnaire surveys,clinical practice experience,the initial draft of the intervention strategies was developed.The intervention strategies were revised and refined through two rounds of Delphi expert consultation.Results The effective response rates for the two rounds of expert consultation questionnaires were 95%and 100%,respectively.The expert authority coefficient was 0.91.The Kendall's coefficient of concordance for the importance of the first and second rounds of expert consultation was 0.224 and 0.202,while the Kendall's coefficient of concordance for feasibility was 0.172 in the second round of expert consultation.The differences were statistically significant(P﹤0.05).The coefficient of variation of entries was 0.00-0.23.Based on the experts'feedback,adjustments were made to several items,resulting in a final intervention strategy that includes 3 primary indicators,9 secondary indicators and 41 intervention measures.Conclusion This strategy demonstrates a high level of scientific rigor and practicality,making it applicable in clinical settings to enhance medication adherence among chronic disease patients effectively,thus providing robust support for patient management.

Objective:To explore the impact of adolescent mental health status on smartphone addiction, and construct a predictive model for smartphone addiction based on the eXtreme Gradient Boosting(XGBoost) algorithm and multivariate Logistic regression.Methods:In April 2023, a cross-sectional survey was conducted among 14 666 adolescents.All participants were systematically evaluated using a self-developed general information questionnaire, the middle school student mental health scale(MSSMHS), the adolescents self-harm scale(ASHS), the interaction anxiousness scale(IAS), the mobile phone addiction index(MPAI), the middle school students shame scale(MSSS), the UCLA loneliness scale(UCLA-LS), the multidimensional peer victimization scale(MPVS), and the basic psychological needs scale(BPNS).R software version 4.3.2 was used for data analysis. Participants were randomly divided into training set and validation set at the ratio of 7∶3.The XGBoost model and multivariate logistic regression model were constructed to predict the risk of smartphone addiction, and a nomogram was plotted.Model performance was evaluated using the Hosmer-Lemeshow test, area under the curve(AUC), and accuracy(ACC).Results:(1) A total of 14 036 high school students were included in the study, with 5 069(36.1%) exhibited smartphone addiction.The training set comprised 9 826 students, with 3 549(36.1%) being smartphone addicts.The validation set included 4 210 students, with 1 520(36.1%) being smartphone addicts.(2) The XGBoost model identified shame-proneness and social anxiety as the two main predictors of smartphone addiction.(3) Multivariate Logistic regression analysis revealed that anxiety( B=0.328, OR(95% CI)=1.39(1.07-1.81), P=0.015), interpersonal sensitivity( B=0.311, OR(95% CI)=1.36(1.05-1.77), P=0.018), learning pressure( B=0.606, OR(95% CI)=1.83(1.46-2.31), P<0.001), mood swings( B=0.775, OR(95% CI)=2.17(1.70-2.78), P<0.001), social anxiety( B=0.024, OR(95% CI)=1.02(1.01-1.04), P<0.001), shame-proneness( B=0.049, OR(95% CI)=1.05(1.04-1.06), P<0.001), and peer victimization( B=0.037, OR(95% CI)=1.04(1.02-1.06), P<0.001) were significant predictors of smartphone addiction.(4) The ACC and AUC values of the XGBoost model were 0.890 and 0.929 in the training set, and 0.865 and 0.864 in the validation set, respectively.The multivariate Logistic regression model achieved ACC and AUC values of 0.870 and 0.854 in the training set, and 0.867 and 0.859 in the validation set, respectively. Conclusion:Anxiety, interpersonal sensitivity, learning pressure, mood swings, social anxiety, shame-proneness, and peer victimization are identified risk predictors of smartphone addiction in high school adolescents.