Background:Duodenal papillary adenoma is a benign tumor with relatively low incidence but significant carcinogenesis potential.Despite the minimal invasiveness and low complication rate,endoscopic papillectomy is associated with a definite risk of recurrence for duodenal papillary adenoma.Investigating the driver genes of duodenal papillary adenoma and establishing predictive models for recurrence and malignant progression could facilitate the precision medicine.Aims:To identify the key driver genes for tumor occurrence,carcinogenesis and recurrence in duodenal papillary adenoma by integrating multi-dimensional bioinformatics approaches based on transcriptomics data,and validate clinically.Methods:Expression profiles of duodenal papillary adenoma and adenocarcinoma were obtained from the GEO database(including data sets GSE189035,GSE94919,GSE111156,and GSE102208).Differentially expressed genes(DEGs)between adenomatous and normal tissues were screened.Weighted gene co-expression network analysis(WGCNA)and pseudo-time analysis were combined to identify the core genes exhibiting an"initial rise followed by decline"expression pattern during the dynamic progression from normal tissue to adenoma and adenocarcinoma.Functional annotation,immune microenvironment profiling,and protein-protein interaction network analysis were performed to explore the tumor-promoting mechanisms of these core genes.Clinical validation was conducted using immunohistochemistry to estimate the gene expression level and its relationship with tumor recurrence.Results:A total of 469 common DEGs were identified.WGCNA revealed that the blue module(including 1 051 genes)was associated with adenoma development and progression(Cor=-0.29,0.15,and 0.11 for normal tissue,adenoma,and adenocarcinoma,respectively).Intersection with DEGs pinpointed four key genes:SLC12A2,BEST4,SLC37A2,and SOAT2.Pseudo-time analysis demonstrated that only SLC12A2 maintained sustained high expression in both adenoma and adenocarcinoma tissues.KEGG enrichment analysis indicated that SLC12A2 was linked to various malignant pathways(e.g.,PD-1/PD-L1 signaling pathway),and its high expression correlated with the reduced immune cell infiltration(e.g.,γδ T cells,CD8+T cells,etc.).Clinical validation by immunohistochemistry confirmed the trend of initial upregulation and subsequent downregulation of SLC12A2 expression in normal,adenoma,and adenocarcinoma tissues.Patients with tumor recurrence showed higher SLC12A2 expression level(P=0.004);likewise,SLC12A2 high expression was associated with an elevated recurrence risk(P=0.034).Conclusions:SLC12A2 serves as a critical driver of tumorigenesis and progression for duodenal papillary adenoma,and might be a promising biomarker for recurrence prediction.

Objective:To investigate the protective effects of resveratrol (RSV) on human sperm cryopreservation and explore its underlying protective mechanisms.Methods:A total of 165 normal fresh semen samples were collected from the Reproductive Medicine Center, Department of Obstetrics and Gynecology and Human Sperm Bank of the First Affiliated Hospital of Anhui Medical University between December 2022 and December 2024. Among them, 65 samples were used to obtain semen parameters before and after conventional freezing. Each sample of the other 104 samples was mixed at a 2∶1 volume ratio with cryoprotectant containing 0, 10 -?, 10 -?, or 10 -? mol/L RSV, followed by cryopreservation in liquid nitrogen for 24 h. Post-thaw assessments included routine sperm parameters, sperm DNA fragmentation index (DFI) evaluated by sperm chromatin dispersion assay, reactive oxygen species (ROS) levels measured via flow cytometry, RSV and nuclear factor erythroid 2-related factor 2 (NRF2) interactions examined by molecular docking and cellular thermal shift assay (CETSA), NRF2 protein contents analyzed by immunofluorescence and Western blotting, mRNA levels of NRF2 and downstream antioxidant proteins Heme Oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO1) quantified by qRT-PCR and effects of NRF2 inhibitor ML385 on sperm parameters. Results:Compared with fresh samples, conventional cryopreservation significantly reduced sperm motility (all P<0.001). The addition of 10 -? mol/L RSV significantly improved the percentage of forward motile sperm after freezing (26.98%±8.98% vs. 19.61%±8.03%, P<0.001) while reducing DFI (9.84%±3.81% vs. 15.06%±4.22%, P<0.001) and ROS levels ( P<0.001) compared with the post-freezing group without the addition of RSV. Both molecular docking analysis and CETSA confirmed that RSV interacted with NRF2. Notably, sperm cryopreserved with 10 -? mol/L RSV exhibited significantly higher contents of NRF2 and its downstream effectors HO-1 and NQO1 compared with the post-freezing group without the addition of RSV (all P<0.001). This protective effect was markedly attenuated by co-treatment with the NRF2 inhibitor ML385, as evidenced by a significant decline in sperm motility ( P<0.001). Conclusion:RSV exerts cryoprotective effects likely through NRF2-mediated antioxidant pathways, reducing oxidative stress and enhancing post-thaw sperm quality.

Background:Duodenal papillary adenoma is a benign tumor with relatively low incidence but significant carcinogenesis potential.Despite the minimal invasiveness and low complication rate,endoscopic papillectomy is associated with a definite risk of recurrence for duodenal papillary adenoma.Investigating the driver genes of duodenal papillary adenoma and establishing predictive models for recurrence and malignant progression could facilitate the precision medicine.Aims:To identify the key driver genes for tumor occurrence,carcinogenesis and recurrence in duodenal papillary adenoma by integrating multi-dimensional bioinformatics approaches based on transcriptomics data,and validate clinically.Methods:Expression profiles of duodenal papillary adenoma and adenocarcinoma were obtained from the GEO database(including data sets GSE189035,GSE94919,GSE111156,and GSE102208).Differentially expressed genes(DEGs)between adenomatous and normal tissues were screened.Weighted gene co-expression network analysis(WGCNA)and pseudo-time analysis were combined to identify the core genes exhibiting an"initial rise followed by decline"expression pattern during the dynamic progression from normal tissue to adenoma and adenocarcinoma.Functional annotation,immune microenvironment profiling,and protein-protein interaction network analysis were performed to explore the tumor-promoting mechanisms of these core genes.Clinical validation was conducted using immunohistochemistry to estimate the gene expression level and its relationship with tumor recurrence.Results:A total of 469 common DEGs were identified.WGCNA revealed that the blue module(including 1 051 genes)was associated with adenoma development and progression(Cor=-0.29,0.15,and 0.11 for normal tissue,adenoma,and adenocarcinoma,respectively).Intersection with DEGs pinpointed four key genes:SLC12A2,BEST4,SLC37A2,and SOAT2.Pseudo-time analysis demonstrated that only SLC12A2 maintained sustained high expression in both adenoma and adenocarcinoma tissues.KEGG enrichment analysis indicated that SLC12A2 was linked to various malignant pathways(e.g.,PD-1/PD-L1 signaling pathway),and its high expression correlated with the reduced immune cell infiltration(e.g.,γδ T cells,CD8+T cells,etc.).Clinical validation by immunohistochemistry confirmed the trend of initial upregulation and subsequent downregulation of SLC12A2 expression in normal,adenoma,and adenocarcinoma tissues.Patients with tumor recurrence showed higher SLC12A2 expression level(P=0.004);likewise,SLC12A2 high expression was associated with an elevated recurrence risk(P=0.034).Conclusions:SLC12A2 serves as a critical driver of tumorigenesis and progression for duodenal papillary adenoma,and might be a promising biomarker for recurrence prediction.

Objective Systematic identification and analysis of the cytochrome P450(CYP450)gene family members and their functions in Chrysanthemum indicum L.to provide a basis for further studies on their roles in the flavonoid synthesis pathway.Methods Based on the genomic data of a diploid Chrysanthemum indicum L.,the members of the P450 gene family were identified and analyzed using bioinformatics methods.Moreover,the transcriptome and quasi-targeted metabolome analysis were combined to screen the flavone synthase Ⅱ(FNSII)genes from the CYP93 family,and their expression levels were tested in different tissues of C.indicum using real-time fluorescence quantitative PCR(qPCR).Results A total of 460 P450 genes were identified from the C.indicum genome,belonging to 43 families within 8 family clans.The encoded proteins ranged from 336 to 1538 amino acids in length,with a relative molecular mass between 38.01 to 175.00 kDa,and an isoelectric point between 5.61 to 9.71.Chromosome localization analysis indicated that these 460 P450 genes were unevenly distributed on nine chromosomes,with the highest number of light-responsive elements in the promoter regions.Expression pattern analysis showed that the expression levels of two FNSII genes(CindChr2G00102820.1 and CindChr8G00552890.1)were significantly higher in leaf of C.indicum than in flower.Conclusion The comprehensive analysis of the P450 gene family in C.indicum is helpful to further elucidate the function of the P450 gene family in flavonoid biosynthesis.

Objective:To investigate the protective effects of resveratrol (RSV) on human sperm cryopreservation and explore its underlying protective mechanisms.Methods:A total of 165 normal fresh semen samples were collected from the Reproductive Medicine Center, Department of Obstetrics and Gynecology and Human Sperm Bank of the First Affiliated Hospital of Anhui Medical University between December 2022 and December 2024. Among them, 65 samples were used to obtain semen parameters before and after conventional freezing. Each sample of the other 104 samples was mixed at a 2∶1 volume ratio with cryoprotectant containing 0, 10 -?, 10 -?, or 10 -? mol/L RSV, followed by cryopreservation in liquid nitrogen for 24 h. Post-thaw assessments included routine sperm parameters, sperm DNA fragmentation index (DFI) evaluated by sperm chromatin dispersion assay, reactive oxygen species (ROS) levels measured via flow cytometry, RSV and nuclear factor erythroid 2-related factor 2 (NRF2) interactions examined by molecular docking and cellular thermal shift assay (CETSA), NRF2 protein contents analyzed by immunofluorescence and Western blotting, mRNA levels of NRF2 and downstream antioxidant proteins Heme Oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO1) quantified by qRT-PCR and effects of NRF2 inhibitor ML385 on sperm parameters. Results:Compared with fresh samples, conventional cryopreservation significantly reduced sperm motility (all P<0.001). The addition of 10 -? mol/L RSV significantly improved the percentage of forward motile sperm after freezing (26.98%±8.98% vs. 19.61%±8.03%, P<0.001) while reducing DFI (9.84%±3.81% vs. 15.06%±4.22%, P<0.001) and ROS levels ( P<0.001) compared with the post-freezing group without the addition of RSV. Both molecular docking analysis and CETSA confirmed that RSV interacted with NRF2. Notably, sperm cryopreserved with 10 -? mol/L RSV exhibited significantly higher contents of NRF2 and its downstream effectors HO-1 and NQO1 compared with the post-freezing group without the addition of RSV (all P<0.001). This protective effect was markedly attenuated by co-treatment with the NRF2 inhibitor ML385, as evidenced by a significant decline in sperm motility ( P<0.001). Conclusion:RSV exerts cryoprotective effects likely through NRF2-mediated antioxidant pathways, reducing oxidative stress and enhancing post-thaw sperm quality.

The female reproductive system is essential for sustaining reproductive endocrine homeostasis,however,its vulnerability to various endogenous and exogenous insults,including pathological conditions,pharmacological agents,genetic predispositions,and environmental factors,often results in compromised fertility.The existing protective approaches(including surgical interventions,hormonal replacement therapies,and assisted reproductive techniques)are constrained by several limitations,such as adverse therapeutic effects,technical complexities,and their incapacity to reverse ovarian senescence.Artemisinin and its derivatives(ARTs),characterized by their unique endoperoxide bridge configuration,have exhibited outstanding therapeutic performance across multiple domains including malaria treatment,anticancer therapy,inflammation modulation,and parasitic infection control.Emerging research has identified their novel protective capabilities against various reproductive system pathologies.This comprehensive review systematically elucidates the molecular mechanisms underlying artemisinin-based interventions in reproductive pathologies and evaluates their clinical translation prospects,thereby proposing innovative strategies for the development of next-generation fertility-protective agents with enhanced safety and efficacy profiles.

BACKGROUND:The imbalance between proliferation and apoptosis of chondrocytes plays an important role in the occurrence and development of osteoarthritis. Previous studies have found that hsa-miR-3202 is involved in regulating the proliferation and apoptosis of various cells. However,no studies have explored the correlation between hsa-miR-3202 and osteoarthritis.OBJECTIVE:To investigate the expression of hsa-miR-3202 in osteoarthritic chondrocytes and its effect on the proliferation and apoptosis of chondrocytes. METHODS:(1) MicroRNAs differentially expressed in osteoarthritic chondrocytes were screened by biogenic analysis. Based on the current research situation at home and abroad,hsa-miR-3202 was selected for follow-up studies,and its target genes were predicted by gene ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. (2) Human normal chondrocyte cell lines C28/I2 in logarithmic growth phase were selected and randomly divided into four groups for culture:in normal group,cells were cultured in normal medium for 24 hours,the medium was then changed to normal medium for another 6 hours of culture,and changed to normal medium for subsequent culture;in lipopolysaccharide group,cells were cultured in lipopolysaccharide-containing medium for 24 hours,the medium was then changed to normal medium for another 6 hours,and changed to normal medium for subsequent culture;in lipopolysaccharide+NC group,cells were cultured in lipopolysaccharide-containing medium for 24 hours,and then transfected with has-miR-3202 mimics control for 6 hours,and the medium was change to normal medium for subsequent culture;in lipopolysaccharide+hsa-miR-3202 mimics group,cells were cultured in lipopolysaccharide-containing medium for 24 hours and then transfected with has-miR-3202 mimics for 6 hours,and the medium was changed to normal medium for subsequent culture. After further 48 hours of culture,the expression level of hsa-miR-3202 was detected by fluorescence quantitative PCR and cell apoptosis was detected by flow cytometry. After further culture of 0-72 hours,cell proliferation activity was detected by cell counting kit-8. RESULTS AND CONCLUSION:Bioinformatics analysis results indicated that hsa-miR-3202 was significantly down-regulated in osteoarthritic chondrocytes. GO functional enrichment and KEGG pathway enrichment showed that the function of hsa-miR-3202 target gene was closely related to cell growth and apoptosis. The results of in vitro cell experiments showed that compared with the normal group,the expression level of hsa-miR-3202 and proliferation ability of chondrocytes were significantly decreased in the lipopolysaccharide group (P<0.05),while the apoptotic rate was significantly increased (P<0.05). Compared with the lipopolysaccharide group,the expression level of hsa-miR-3202 and proliferation ability of chondrocytes were significantly increased in the lipopolysaccharide+hsa-miR-3202 mimics group (P<0.05),while the apoptotic rate was significantly decreased (P<0.05). To conclude,the expression of hsa-miR-3202 is down-regulated in osteoarthritic chondrocytes to inhibit cell proliferation and promote cell apoptosis,thus affecting the occurrence and development of osteoarthritis.

Objective:To explore the guiding significance of Expanded Carrier Screening (ECS) for the fertility process of individuals undergoing Assisted Reproductive Technology (ART) in Dalian.Methods:A retrospective analysis was conducted on the ECS results of patients who visited the Dalian Maternal and Child Health Care Center (Group) from January 2023 to June 2024. The study included the screening of 155 monogenic diseases and the ART status of carrier couples.Results:Among the 1 384 patient samples, 490 carriers were identified, yielding a detection rate of 35.41% (490/1 384). A total of 100 diseases and 91 pathogenic genes were detected. The most frequently identified disease was autosomal recessive hearing loss type 4 (SLC26A4), with a carrier rate of 3.54% (49/1384). For the 490 carriers, their partners were recalled for testing and, 322 partners participated. Eight high-risk couples were identified, accounting for 2.48% (8/322). Among the 322 couples, 4 used artificial insemination by husband (AIH, 1.24%, 4/322), 166 used in vitro fertilization (IVF, 51.55%, 166/322), 139 used intracytoplasmic sperm injection (ICSI, 43.17%, 139/322), and 1 underwent preimplantation genetic testing for monogenic diseases (PGT-M, 0.31%, 1/322). Twelve couples (3.73%, 12/322) did not undergo ART, were not pregnant, or chose gamete donation. Among the eight high-risk couples, three underwent IVF, four underwent ICSI, and one underwent PGT-M. One couple conceived through ICSI, and the fetus was followed up. Amniotic fluid Sanger sequencing revealed that the fetus had compound heterozygous mutations in the PAH gene, with two suspected pathogenic variants at c.532G>A and c.1174T>A. One couple undergoing PGT-M currently has one retrieved oocyte and zero usable blastocysts. The embryo carries a paternal mutation and is aneuploid.Conclusions:This study not only identified the common pathogenic diseases in Dalian, providing a reference for clinical treatment, but also validated the critical significance of ECS for individuals undergoing ART.

Objective:To examine the validity and reliability of the DSM-5 Social Anxiety Disorder Severity Scale(SAD-D)in a Chinese adult population.Methods:The Chinese version of the DSM-5 Social Anxiety Disor-der Severity Scale was administered via online data collection platform Credamo to 300 adults(Sample 1,for item analysis,exploratory factor analysis and item selection of brief version of SAD-D)and 528 adults(Sample 2,for confirmatory factor analysis,criterion validity test,measurement invariance analysis and internal consistency reliabil-ity analysis for both SAD-D and its brief version).Criterion validity was tested with the Social Phobia Scale(SPIN)and Personal Report of Confidence as a Speaker(PRCS).A brief version of the scale was developed by u-sing the Ant Colony Optimization(ACO).A retest was conducted with 152 participants from Sample 2 after three weeks.Results:Exploratory factor analysis indicated that the SAD-D was a unidimensional scale with factor load-ings ranging from 0.49 to 0.82,and the results of the confirmatory factor analysis also supported the unidimension-al structure(x2/df=3.49,RMSEA=0.069,CFI=0.971,TLI=0.962,SRMR=0.028).The scores of Chinese version of the SAD-D were positively correlated with the SPIN scores(ICC=0.70,P＜0.001)and PRCS scores(ICC=0.73,P＜0.001).The Cronbach'α of the scale was 0.92,and the retest reliability was 0.85.The scale dem-onstrated cross-gender measurement invariance(△CFI＜0.01,△RMSEA＜0.01).The brief version of the SAD-D was selected as items 2,5,and 6,and its Cronbach'α coefficient was 0.86.Conclusion:The Chinese version of the SAD-D has satisfactoryvalidity andreliability,making it suitable for the assessment of social anxiety symptoms with Chinese adults.

Objective:To examine the validity and reliability of the DSM-5 Social Anxiety Disorder Severity Scale(SAD-D)in a Chinese adult population.Methods:The Chinese version of the DSM-5 Social Anxiety Disor-der Severity Scale was administered via online data collection platform Credamo to 300 adults(Sample 1,for item analysis,exploratory factor analysis and item selection of brief version of SAD-D)and 528 adults(Sample 2,for confirmatory factor analysis,criterion validity test,measurement invariance analysis and internal consistency reliabil-ity analysis for both SAD-D and its brief version).Criterion validity was tested with the Social Phobia Scale(SPIN)and Personal Report of Confidence as a Speaker(PRCS).A brief version of the scale was developed by u-sing the Ant Colony Optimization(ACO).A retest was conducted with 152 participants from Sample 2 after three weeks.Results:Exploratory factor analysis indicated that the SAD-D was a unidimensional scale with factor load-ings ranging from 0.49 to 0.82,and the results of the confirmatory factor analysis also supported the unidimension-al structure(x2/df=3.49,RMSEA=0.069,CFI=0.971,TLI=0.962,SRMR=0.028).The scores of Chinese version of the SAD-D were positively correlated with the SPIN scores(ICC=0.70,P＜0.001)and PRCS scores(ICC=0.73,P＜0.001).The Cronbach'α of the scale was 0.92,and the retest reliability was 0.85.The scale dem-onstrated cross-gender measurement invariance(△CFI＜0.01,△RMSEA＜0.01).The brief version of the SAD-D was selected as items 2,5,and 6,and its Cronbach'α coefficient was 0.86.Conclusion:The Chinese version of the SAD-D has satisfactoryvalidity andreliability,making it suitable for the assessment of social anxiety symptoms with Chinese adults.