Objective This study aims to elucidate the functional mechanism through which seminal plasma exosomal miR-26a-5p and its target genes contribute to idiopathic teratozoospermia,with emphasis on their regulatory pathways in sperm morphogenesis defects,thereby establishing a theoretical foundation for novel diagnostic markers and targeted therapies.Methods A case-control study design was adopted,enrolling 154 male subjects(84 in the teratozoospermia group[TZS]and 70 in the normal control group[NC]).Seminal plasma exosomes were isolated to screen differentially expressed miRNAs,followed by prediction and analysis of target genes and signaling pathways.Dual-luciferase reporter gene assays were employed to validate the direct binding of miR-26a-5p to PTEN.Quantitative real-time PCR and Western blot were used to measure miRNA and protein expression levels.Spearman correlation analysis evaluated relationships between miR-26a-5p,PTEN,and sperm morphological parameters,while ROC curve analysis assessed diagnostic efficacy.Results Seminal plasma exosomal miRNA sequencing identified 11 differentially expressed miRNAs,with miR-26a-5p significantly upregulated in the TZS group(P<0.05)and negatively correlated with the percentage of normal sperm morphology in TZS(r=-0.762,P<0.05).PTEN was confirmed as a direct target of miR-26a-5p,with its expression significantly reduced in the TZS group(P<0.05)and positively correlated with normal sperm morphology(r=0.821,P<0.05).A negative correlation was observed between miR-26a-5p and PTEN(r=-0.878,P<0.05).Dual-luciferase assays demonstrated that miR-26a-5p specifically inhibited PTEN 3'UTR activity(45%reduction in luciferase activity,P<0.05).Functional enrichment analysis revealed that the miR-26a-5p-PTEN axis exacerbates oxidative stress,DNA damage,and abnormal spermatocyte division by regulating the PI3K/AKT/mTOR,p53,Wnt/β-catenin,and NF-κB pathways.ROC analysis showed diagnostic efficacy for teratozoospermia with AUCROC values of 0.785(sensitivity 78.9%,specificity 71.3%)for miR-26a-5p and 0.754(sensitivity 73.3%,specificity 75.1%)for PTEN.Conclusions miR-26a-5p suppresses PTEN through targeted inhibition,activating PI3K/AKT/mTOR signaling pathways while impairing DNA repair functions.This cascade leads to accumulated oxidative stress and sperm morphological abnormalities.The combined pattern of elevated seminal plasma exosomal miR-26a-5p expression and reduced PTEN levels demonstrates diagnostic potential for idiopathic teratozoospermia,with its molecular mechanism providing a theoretical foundation for biomarker development.Targeted silencing of miR-26a-5p,either alone or in combination with AKT/mTOR inhibitors and antioxidant therapy,may represent a novel strategy for improving sperm morphology.

Objective:To investigate the role and molecular mechanism of dual-specificity phosphatase 26(DUSP26)in the proliferation,migration and invasion of lung adenocarcinoma(LUAD)A549 cells.Methods:The expression profile of DUSP26 was retrieved from the GEPIA2 tumor database,and its differential expression in LUAD tissues and normal human lung tissues were analyzed.Twelve pairs of LUAD tissue and paracancerous tissue surgically resected at Pingxiang People's Hospital between October 2022 and October 2023 were collected.The difference in DUSP26 expression between LUAD tissues and paracancerous tissues was analyzed using immunohistochemical(IHC)staining and Western blotting(WB).Additionally,the expression of DUSP26 in four LUAD cells(A549,SK-LU-1,Calu-3,H1299)and two normal bronchial epithelial cells(BEAS-2B,HBEC)was detected using WB method.A549 cells stably overexpressing DUSP26(DUSP26-OE)or corresponding negative control(DUSP26-OENC)were constructed via lentiviral transfection.The effects of DUSP26 overexpression on cell proliferation,migration and invasion were detected using colony formation,scratch assay,and Transwell chamber assay,respectively.The expression levels of TGF-β1/SMAD2/3 pathway-and epithelial-mesenchymal transition(EMT)-related proteins were detected using WB method,and the expression levels of Ki-67 and cyclin D1 in cells were detected by immunofluorescence staining.Rescue experiments were conducted by adding 5 ng/mL recombinant TGF-β1.A nude mouse xenograft model was established using A549 cells to observe the effect of DUSP26 overexpression on the in vivo growth of transplanted tumors.The expression levels of TGF-β1/SMAD2/3 pathway-and EMT-related proteins in transplanted tumor tissues were assessed using WB method,and the expression levels of Ki-67 and cyclin D1 in transplanted tumor tissues were detected using immunofluorescence staining.Results:DUSP26 expression was downregulated in both LUAD tissues and cells(P<0.05,P<0.01,P<0.001 or P<0.000 1).Compared with the DUSP26-OENC group,the DUSP26-OE group showed significantly reduced proliferation,migration and invasion of A549 cells(P<0.01 or P<0.001).Furthermore,the protein levels of TGF-β1,p-SMAD2/3,vimentin,N-cadherin,snail,Ki-67,and cyclin D1 were significantly reduced(P<0.01,P<0.001 or P<0.000 1),while E-cadherin level was increased in the DUSP26-OE group(P<0.000 1).The addition of 5 ng/mL TGF-β1 recombinant protein partially reversed the effects of DUSP26 overexpression in vitro experiments.The nude mice A549 cell xenograft model was successfully constructed.The growth rate of transplanted tumors was significantly slower in the DUSP26-OE group,with reduced volume and mass(all P<0.001).The protein levels of TGF-β1,p-SMAD2/3,vimentin,N-cadherin,snail,Ki-67,and cyclin D1 in the transplanted tumor tissues were all reduced(P<0.01 or P<0.001),while E-cadherin level was increased(P<0.000 1).Conclusion:DUSP26 is downregulated in both LUAD tissues and cells.Upregulation of DUSP26 suppresses the proliferation,migration and invasion of A549 cells by inhibiting the TGF-β1/SMAD2/3 signaling pathway.

Objective To explore the clinical value of mini-incision combined with balloon dilatation under the guidance of miniprobe echoendoscope in the treatment of refractory benign esophageal stenosis.Methods From October 2019 to October 2023,30 cases of esophageal early cancer with esophageal scar stenosis after endoscopic submucosal dissection(ESD)were selected and randomly divided into study group(miniprobe echoendoscope guided small incision+balloon dilatation,15 cases)and control group[esophageal radial incision(ERI),15 cases],EUS in the study group judged the hierarchical structure of esophageal wall before operation,and excluded those with esophageal cancer recurrence and submucosal invasive growth.According to miniprobe echoendoscope,the length of scar segment and the scope of scar ring lumen were defined,and the location and length of pre-incision were defined,and then balloon was used to expand step by step.The short-term postoperative complications,Stooler dysphagia scale score and restenosis rate of the two groups were observed,and the feasibility,safety and efficacy were evaluated.Results The incidence of short-term postoperative complications 6.7％in the study group was significantly lower than that 46.7％in the control group,the difference was statistically significant(P＜0.05),and the Stooler dysphagia scale score were(1.133±0.990)and(1.600±0.737)in the study group at 1 and 2 months after operation with significantly lower than those(2.067±1.033)and(2.467±0.915)in the control group,the differences were statistically significant(P＜0.05).Two months after operation,the restenosis rate was 13.3％,which was significantly lower than that 46.7％of the control group,the difference was statistically significant(P＜0.05),and the scar thickness under miniprobe echoendoscope(2.113±0.887)mm was significantly thinner than that(3.353±1.468)mm of the control group,the difference was statistically significant(P＜0.05).Conclusion Preoperative miniprobe echoendoscope evaluation,can accurately locate lesions,endoscopic mini-incision combined with balloon mechanical expansion are safe and effective,with less pain and fewer postoperative complication.

Objective To explore the clinical value of mini-incision combined with balloon dilatation under the guidance of miniprobe echoendoscope in the treatment of refractory benign esophageal stenosis.Methods From October 2019 to October 2023,30 cases of esophageal early cancer with esophageal scar stenosis after endoscopic submucosal dissection(ESD)were selected and randomly divided into study group(miniprobe echoendoscope guided small incision+balloon dilatation,15 cases)and control group[esophageal radial incision(ERI),15 cases],EUS in the study group judged the hierarchical structure of esophageal wall before operation,and excluded those with esophageal cancer recurrence and submucosal invasive growth.According to miniprobe echoendoscope,the length of scar segment and the scope of scar ring lumen were defined,and the location and length of pre-incision were defined,and then balloon was used to expand step by step.The short-term postoperative complications,Stooler dysphagia scale score and restenosis rate of the two groups were observed,and the feasibility,safety and efficacy were evaluated.Results The incidence of short-term postoperative complications 6.7％in the study group was significantly lower than that 46.7％in the control group,the difference was statistically significant(P＜0.05),and the Stooler dysphagia scale score were(1.133±0.990)and(1.600±0.737)in the study group at 1 and 2 months after operation with significantly lower than those(2.067±1.033)and(2.467±0.915)in the control group,the differences were statistically significant(P＜0.05).Two months after operation,the restenosis rate was 13.3％,which was significantly lower than that 46.7％of the control group,the difference was statistically significant(P＜0.05),and the scar thickness under miniprobe echoendoscope(2.113±0.887)mm was significantly thinner than that(3.353±1.468)mm of the control group,the difference was statistically significant(P＜0.05).Conclusion Preoperative miniprobe echoendoscope evaluation,can accurately locate lesions,endoscopic mini-incision combined with balloon mechanical expansion are safe and effective,with less pain and fewer postoperative complication.

In order to establish a serological method for the detection of bovine parainfluenza virus type 3(BPIV3),the prokaryotic expression and purification of BPIV3 HN,NP,F,and P proteins were carried out,and the optimal protein-coated antigen was screened,and an indirect ELISA de-tection method was established.The results showed that the four recombinant proteins of BPIV3,rHN,rNP,rF,and rP were expressed,and the checkerboard titration results showed that rHN pro-tein had the highest P/N value as the coating protein,so it was used for the subsequent method es-tablishment.The optimal reaction conditions for indirect ELISA were found to be:the mass con-centration of the antigen coating was 0.5 mg/L,37 ℃ 1.5 h,5％skim milk,overnight blocking at 4 ℃,serum dilution at 1∶50,incubation at 37 ℃ 1 h,secondary antibody dilution at 1∶10 000 and incubation at 37℃ 0.5 h,substrate reaction conditions were 37℃ for 12 min.The results of speci-ficity experiments showed that the established method could specifically identify BPIV3 antibody-positive serum with a sensitivity of 1∶800,and the coefficient of variation in the detection of intra-and inter-assay repeatability was less than 10％,and the overall coincidence rate of the same batch of samples detected with the SVANOVIR kit was 92.22％.This method was used to detect 192 se-rum samples in Hebei Province,and the positive rate of BPIV3 antibody in serum was 66.15％.The indirect ELISA detection method of BP1V3 antibody constructed in this study is suitable for large-scale clinical serological investigations,and provides valuable data support for the research and de-velopment of BPIV3 antigen and antibody detection kits in China.

Objective To investigate the effect of fumigation decoction on the expression of aggrecan(AGG)and type Ⅱ collagen(COL2A1)in an intervertebral disc degeneration(IDD)model,and to gain an in-depth understanding of the cellular and molecular mechanisms underlying traditional Chinese medicine fumigation therapy.Methods An IDD model was established using Sprague-Dawley rats by inducing disc injury via needle puncture.The rats were divided into three groups:normal control(CON),model(PUN),and fumigation(CHF).Only the CHF group received treatment with fumigation decoction for 4 weeks.Imaging analysis was conducted to assess the relative height of intervertebral discs,while Masson's trichrome staining was used to observe structural changes in the discs.The expression of NLRP3 inflammasome was detected by immunohistochemistry.Quantitative RT-PCR and Western blotting were employed to evaluate the expression levels of AGG and COL2A1 genes and proteins.Results A total of 12 rats were randomly divided into 3 groups,with 4 rats in each group.Compared with the CON group,disc height was significantly lost in the PUN group(P＜0.05),and disc height could be partially restored in the CHF group(P＜0.05).In the PUN group,the expression levels of AGG mRNA and COL2A1 mRNA and protein in the nucleus pulposus tissue were significantly lower than those in the CON group(P＜0.05).Compared with the PUN group,the CHF group showed significantly increased expression levels of AGG mRNA and COL2A1 mRNA and protein in the nucleus pulposus tissue(P＜0.05).Histological analysis revealed that,compared with the PUN group,the CHF group exhibited partial restoration of the gel-like properties of the nucleus pulposus and an increase in proteoglycan content.Additionally,NLRP3 expression in the nucleus pulposus tissue of the CHF group was markedly reduced.Conclusion Fumigation decoction treatment can enhance the expression of AGG and COL2A1 and decrease the expression of NLRP3 in IDD models,which may be one of its mechanisms for treating lumbar degenerative diseases.This research provides new insights into the treatment of IDD.

Objectives:To investigate the effect and mechanism of mitochondrial targeting peptide SS-31 on senescence and pyroptosis of nucleus pulposus(NP)cells induced by inflammation.Methods:NP cells were isolated and cultured in vitro from 6-week-old male SD rats,and were divided into control group,lipopolysaccharide(LPS)group,and LPS+SS-31 group;And the control group of cells received no treatment,LPS group of cells were treated with 100μg/mL LPS for 24h,and LPS+SS-31 group of cells were pre-treated with SS-31 30min before LPS intervention.The levels of cellular senescence and pyroptosis were assessed in each group using β-galactosidase(SA-β-Gal)staining and westem blot(WB)assay;Transmission electron mi-croscopy and oxygen consumption rate(OCR)assay were used to analyze the mitochondrial function of NP cells in each group;The expressions of phosphorylated nuclear factor-κB-p65(p-NF-κB-p65),cyclic GMP-AMP synthase(cGAS),and interferon stimulating gene(STING)in each group were detected by WB and im-munofluorescence;Hematoxylin-eosin(HE)staining,immunohistochemistry(IHC)staining and WB assay were performed to analyze the effects of in vivo injection of SS-31 on intervertebral disc degeneration,senescence,and pyroptosis.Results:The positive SA-β-Gal staining of NP cells,expression of senescence marker pro-teins(p53,p21),and pyroptosis-related molecules(NLRP3,Caspase-1 p20)were significantly lower in the LPS+SS-31 group compared with the LPS group(P＜0.05);Transmission electron microscopy(TEM)showed that the mitochondrial swelling of NP cells was round,the matrix was translucent,and the interstitial lumen of the mitochondrial cristae was dilated in the LPS group,while the mitochondrial swelling was subsided,and the morphology of mitochondria tended to be normalized in the LPS+SS-31 group.Meanwhile,the maximum respi-ration level of NP cells was significantly enhanced in the LPS+SS-31 group compared with the LPS group(P＜0.05).The results of WB and immunofluorescence showed that LPS significantly up-regulated the expression of p-NF-κB-p65,cGAS,and STING(P＜0.05),while SS-31 pretreatment significantly inhibited the expression of p-NF-κB-p65,cGAS and STING in NP cells induced by LPS(P＜0.05).Finally,in vivo experiments con-firmed that local injection of SS-31 reduced the senescence and pyroptosis of NP cells,and alleviated the degeneration of intervertebral disc induced by annulus fibropuncture.Conclusions:SS-31 alleviates inflamma-tion-induced senescence and pyroptosis of NP cells,which is related to the inhibition of NF-κB signaling pathway and cGAS/STING signaling pathway.

Functional constipation is a common functional gastrointestinal disorder with a multifactorial and incompletely understood pathogenesis.Recent studies have revealed that its development involves the interplay of multiple mechanisms,including neurogenic and myogenic dysfunction of the colon,reduction and impairment of interstitial cells of Cajal(ICCs),outlet obstruction,dysregulation of the gut-brain axis,immune activation,and gut microbiota imbalance.Slow-transit constipation is mainly associated with enteric neural abnormalities,disruption of ICC signaling,and inflammation,whereas outlet obstruction constipation often results from pelvic floor dysfunction and rectal hyposensitivity.Dysregulation of the gut-brain axis plays a central role,involving impaired central regulation,hormonal imbalance,and enhanced local immune response.Additionally,gut microbial metabolites such as short-chain fatty acids,bile acids,and methane affect colonic motility and inflammation.This review summarizes the current understanding and research progress on the pathogenesis of functional constipation,providing insights for mechanism-based and individualized therapeutic approaches.

Objective To investigate the status and influencing factors of psychological distress associated with weight loss among young and middle-aged adults with obesity,so as to provide a theoretical basis for developing personalized interventions and improving psychological well-being.Methods A convenience sampling method was used to select young and middle-aged obese patients who visited the weight management clinic of a Grade A tertiary hospital in Jiangxi Province from October 2024 to May 2025.Data were collected using a general information questionnaire,the distress thermometer,the perceived social support scale,the brief illness perception questionnaire,and the simplified coping style questionnaire.Binary logistic regression was applied to identify factors influencing psychological distress.Results A total of 204 valid questionnaires were collected.The detection rate of significant psychological distress was 58.82%(120 cases).Regarding weight loss methods,32.84%of participants opted for medication.The top five issues reported on the psychological distress problem list were:appearance/body image,work/studies,lack of time/energy to care for elderly parents or children,bathing/dressing,and relationship with a partner.Binary logistic regression indicated that age,body mass index,waist circumference,history of chronic disease,perceived social support,illness perception,and negative coping style were significant influencing factors of psychological distress(P﹤0.05).Conclusions The rate of significant psychological distress is relatively high among young and middle-aged obese patients and is influenced by multiple factors.Medical staff may develop personalized interventions based on these factors to reduce the incidence of psychological distress.

Objective This study aims to elucidate the functional mechanism through which seminal plasma exosomal miR-26a-5p and its target genes contribute to idiopathic teratozoospermia,with emphasis on their regulatory pathways in sperm morphogenesis defects,thereby establishing a theoretical foundation for novel diagnostic markers and targeted therapies.Methods A case-control study design was adopted,enrolling 154 male subjects(84 in the teratozoospermia group[TZS]and 70 in the normal control group[NC]).Seminal plasma exosomes were isolated to screen differentially expressed miRNAs,followed by prediction and analysis of target genes and signaling pathways.Dual-luciferase reporter gene assays were employed to validate the direct binding of miR-26a-5p to PTEN.Quantitative real-time PCR and Western blot were used to measure miRNA and protein expression levels.Spearman correlation analysis evaluated relationships between miR-26a-5p,PTEN,and sperm morphological parameters,while ROC curve analysis assessed diagnostic efficacy.Results Seminal plasma exosomal miRNA sequencing identified 11 differentially expressed miRNAs,with miR-26a-5p significantly upregulated in the TZS group(P<0.05)and negatively correlated with the percentage of normal sperm morphology in TZS(r=-0.762,P<0.05).PTEN was confirmed as a direct target of miR-26a-5p,with its expression significantly reduced in the TZS group(P<0.05)and positively correlated with normal sperm morphology(r=0.821,P<0.05).A negative correlation was observed between miR-26a-5p and PTEN(r=-0.878,P<0.05).Dual-luciferase assays demonstrated that miR-26a-5p specifically inhibited PTEN 3'UTR activity(45%reduction in luciferase activity,P<0.05).Functional enrichment analysis revealed that the miR-26a-5p-PTEN axis exacerbates oxidative stress,DNA damage,and abnormal spermatocyte division by regulating the PI3K/AKT/mTOR,p53,Wnt/β-catenin,and NF-κB pathways.ROC analysis showed diagnostic efficacy for teratozoospermia with AUCROC values of 0.785(sensitivity 78.9%,specificity 71.3%)for miR-26a-5p and 0.754(sensitivity 73.3%,specificity 75.1%)for PTEN.Conclusions miR-26a-5p suppresses PTEN through targeted inhibition,activating PI3K/AKT/mTOR signaling pathways while impairing DNA repair functions.This cascade leads to accumulated oxidative stress and sperm morphological abnormalities.The combined pattern of elevated seminal plasma exosomal miR-26a-5p expression and reduced PTEN levels demonstrates diagnostic potential for idiopathic teratozoospermia,with its molecular mechanism providing a theoretical foundation for biomarker development.Targeted silencing of miR-26a-5p,either alone or in combination with AKT/mTOR inhibitors and antioxidant therapy,may represent a novel strategy for improving sperm morphology.