Objective:To observe the effect of exercise-regulated miR-344g-5p on the fibrosis-related SMAD genes and transforming growth factor beta (TGF-β) in the myocardia of mice modeling diabetes.Methods:Twenty-four male C57BL/6 mice (6-8 weeks old) were randomly divided into a control group ( n=12) and a type 2 diabetes group ( n=12). Each group was further divided into two subgroups based on exercise status to form a sedentary control group, an exercise control, a sedentary type 2 diabetes group and an exercise type 2 diabetes group with six mice in each subgroup. The control groups were fed a normal diet, while the type 2 diabetes groups were on a high-fat diet for 12 weeks. Type 2 diabetes was then induced by intraperitoneal injection of streptozotocin. Two weeks later, the exercise groups began 40 minutes of daily swimming training, five days a week for eight consecutive weeks. Right after that, their cardiac function was measured using a small animal ultrasound system and the derived ejection fraction (EF) and the maximal early (E) and late (A) transmitral velocities ratio (E/A ratio) in diastole. They were then sacrificed and myocardial tissue was resected and stained with Sirius red. The expression of miR-344g-5p in the myocardium was detected using quantitative polymerase chain reactions. The expression of phosphorylated SMAD3 (p-SMAD3) and TGF-β were assessed using western blotting. The Target Scan database was exploited to analyze whether there were predicted targets of miR-344g-5p and pro-fibrotic genes such as SMAD3, TGFBR2, COL1A2 and COL12A1, and to determine any correlations in the gene regulation. Results:After 22 weeks, the EF and E/A ratio in the sedentary type 2 diabetes group were (57.5±4.1)% and (1.4±0.3), respectively, both significantly lower than in the other groups. Myocardial collagen fibers in the sedentary type 2 diabetes group were significantly more abundant than in the sedentary control and exercise type 2 diabetes groups. And miR-344g-5p expression in the myocardia of the exercise type 2 diabetes group was significantly greater than that in the sedentary type 2 diabetes group. The expression of p-SMAD3 and TGF-β in the myocardia of the sedentary type 2 diabetes group was significantly higher than in the sedentary control and exercise type 2 diabetes groups. Target Scan analysis revealed that miR-344g-5p had potential binding sites with several fibrosis-related genes such as SMAD3, TGFBR2, COL1A2, and COL12A1. Based on the reduction in TGF-β and p-SMAD protein expression in the exercise type 2 diabetes group, it was hypothesized that miR-344g-5p may inhibit the post-transcriptional processes of those genes.Conclusions:Exercise promotes the recovery of diabetic cardiomyopathy by upregulating myocardial miR-344g-5p expression, which subsequently targets and suppresses p-SMAD3 and TGF-β protein expression, thereby reducing diabetic myocardial fibrosis.

Objective To examine the time window effect of high-load exerciseon skeletal muscle in-flammation genes,and identify the key genes and signaling pathways involved in this process.Methods Forty-eight Sprague-Dawley rats were randomly assigned to a control group(group C,n=8)and an ex-ercise group(group E,n=40).Gastrocnemius muscles were collected immediately(group E0),12h(group E12),24h(group E24),48h(group E48),and 72 h(group E72)after exercise for transcrip-tome sequencing.Differentially expressed genes(DEGs)were identified,and enrichment analysis was carried out using the Gene Ontology(GO)and the Kyoto Encyclopedia of Genes and Genomes(KEGG)annotations.Meanwhile,inflammation-related genes were obtained from databases,and differ-entially expressed inflammation-related genes(DEIRGs)were done through identifying their intersec-tion with DEGs.Moreover,the Mfuzz algorithm was used for time series clustering to obtain subsets with similar characteristics.GO and KEGG analyses,along with protein interaction network analysis,were performed to obtain key DEIRGs,followed by secondary functional enrichment to analyze changes in expression of key genes over time and identify key signaling pathways.Results Seven DEIRG clus-ters were obtained through Mfuzz time series clustering of skeletal muscle inflammation genes after high-load exercise.Overall,the expression of DEGs in cluster 5 was downregulated,while that in cluster 7 was upregulated.The expression of DEGs in clusters 3 and 4 was upregulated at E0 and rap-idly downregulated at E12.In contrast,the expression of DEGs in cluster 2 and 6 were downregulated at E0 and rapidly upregulated at E12.The expression of DEGs in cluster 1 was upregulated at E0,rapidly downregulated at E12,and remained upregulated at E24.Screening identified TP53,STAT3,CD44,AKT1,KDR,GJA1,CYCS,HIF1A,IQGAP3,FASN,and TFRC as key DEIRGswhich were enriched in apoptosis,HIF-1,apoptosis,ferroptosis,MAPK,VEGF,PI3K-Akt,insulin resistance,FoxO,AMPK and the JAK-STAT signaling pathway.Conclusion Inflammation-related genes exhibit temporal dynamic changes in exercise-induced muscle damage and show significant time window effects at 12 h after exercise.The key targets STAT3,AKT1 and HIF-1A react to exercise-induced muscle damage through the JAK-STAT,PI3K-Akt,HIF-1 and VEGF signaling pathways,and promote tissue repair.

AIM:To investigate the mechanism through which aerobic exercise improves inflammatory senes-cence in SAMP8 mice via Janus kinase 3(JAK3)-signal transducer and activator of transcription 5α(STAT5α)signaling pathway and to provide novel insights for anti-inflammatory aging interventions.METHODS:Sixteen 28-week-old SAMP8 mice were randomly divided into model(Mod)group and exercise(Exe)group,with 8 mice in each group.Addi-tionally,8 senile SAMR1 mice served as control(Con)group.The mice in Exe group underwent an 8-week aerobic training regimen on a treadmill.We monitored the changes in hair quality and body weight,immune organ indexes,histomorpho-logical alterations in immune organs,the expression levels of spleen aging marker P16,and the serum and spleen levels of inflammatory factors[tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)]and aging indicator IL-2.Moreover,we examined the alterations in the mRNA and protein expression of IL-2,JAK3 and STAT5α in the spleen.RESULTS:(1)Compared with Con group,the mice in Mod group exhibited decreased hair luster and volume,with some instances of alopecia areata,increased body weight,markedly reduced spleen and thymus indexes,evident atrophy and senescence of immune organs,and markedly decreased concentration of IL-2 in the serum and spleen.The expression of P16 in the spleen,and the serum and spleen levels of inflammatory factors TNF-α and IL-1β were significantly increased.The mRNA expression of IL-2 and IL-2 receptor subunit gamma(IL-2RG)in the spleen was markedly decreased,whereas that of JAK3 and STAT5α was markedly increased.Furthermore,the protein expression of IL-2 in the spleen was markedly de-creased,whereas that of STAT5α and JAK3 was markedly increased.(2)Compared with Mod group,the mice in Exe group showed relatively vigorous hair growth and better hair luster,lower body weight,markedly increased spleen and thy-mus indexes,and delayed immune organ aging.The concentration of IL-2 in the serum and spleen was markedly in-creased,while the expression of P16 in the spleen was markedly decreased.The levels of TNF-α and IL-1β in the serum and spleen were markedly decreased.The mRNA expression of IL-2 and IL-2RG in the spleen was markedly increased,whereas that of JAK3 and STAT5α was markedly decreased.The protein levels of IL-2 were markedly increased,whereas those of STAT5α and JAK3 were markedly decreased.CONCLUSION:Aerobic exercise can delay inflammatory aging in mice possibly through the JAK3-STAT5α signaling pathway.

Breast cancer organoids faithfully reflect the structural and functional characteristics of primary tumors, demonstrating broad application potential. Clinically, organoids have been utilized for drug sensitivity testing, significantly enhancing the precision of personalized treatment strategies. Organoids play a crucial role in accelerating new drug development, uncovering drug resistance mechanisms, and improving the efficacy of immunotherapy. In-depth analysis of the application of breast cancer organoid technology in clinical and transformation can provide new ideas for clinical diagnosis and treatment of breast cancer.

Objective To examine the time window effect of high-load exerciseon skeletal muscle in-flammation genes,and identify the key genes and signaling pathways involved in this process.Methods Forty-eight Sprague-Dawley rats were randomly assigned to a control group(group C,n=8)and an ex-ercise group(group E,n=40).Gastrocnemius muscles were collected immediately(group E0),12h(group E12),24h(group E24),48h(group E48),and 72 h(group E72)after exercise for transcrip-tome sequencing.Differentially expressed genes(DEGs)were identified,and enrichment analysis was carried out using the Gene Ontology(GO)and the Kyoto Encyclopedia of Genes and Genomes(KEGG)annotations.Meanwhile,inflammation-related genes were obtained from databases,and differ-entially expressed inflammation-related genes(DEIRGs)were done through identifying their intersec-tion with DEGs.Moreover,the Mfuzz algorithm was used for time series clustering to obtain subsets with similar characteristics.GO and KEGG analyses,along with protein interaction network analysis,were performed to obtain key DEIRGs,followed by secondary functional enrichment to analyze changes in expression of key genes over time and identify key signaling pathways.Results Seven DEIRG clus-ters were obtained through Mfuzz time series clustering of skeletal muscle inflammation genes after high-load exercise.Overall,the expression of DEGs in cluster 5 was downregulated,while that in cluster 7 was upregulated.The expression of DEGs in clusters 3 and 4 was upregulated at E0 and rap-idly downregulated at E12.In contrast,the expression of DEGs in cluster 2 and 6 were downregulated at E0 and rapidly upregulated at E12.The expression of DEGs in cluster 1 was upregulated at E0,rapidly downregulated at E12,and remained upregulated at E24.Screening identified TP53,STAT3,CD44,AKT1,KDR,GJA1,CYCS,HIF1A,IQGAP3,FASN,and TFRC as key DEIRGswhich were enriched in apoptosis,HIF-1,apoptosis,ferroptosis,MAPK,VEGF,PI3K-Akt,insulin resistance,FoxO,AMPK and the JAK-STAT signaling pathway.Conclusion Inflammation-related genes exhibit temporal dynamic changes in exercise-induced muscle damage and show significant time window effects at 12 h after exercise.The key targets STAT3,AKT1 and HIF-1A react to exercise-induced muscle damage through the JAK-STAT,PI3K-Akt,HIF-1 and VEGF signaling pathways,and promote tissue repair.

Objective To explore the mechanism of Quhan Zhufeng Mixture on proliferation and apoptosis of rheumatoid arthritis fibroblast-like synoviocyte(RA-FLS)based on NDRG2/JAK2/STAT3 signaling pathway.Methods RA-FLS cells were cultured in vitro,and were divided into ① blank serum group,methotrexate group,Quhan Zhufeng Mixture low-,medium-and high-dosage groups;② blank serum group,AG490 group,Quhan Zhufeng Mixture low-,medium-and high-dosage groups.Different concentrations of drug-containing serum were used to intervene cells.Cell proliferation was detected by CCK-8 method,apoptosis was detected by flow cytometry,and mRNA expressions of Bax,Bcl-2,Caspace-3,Caspace-9,N-myc downstream regulatory gene 2(NDRG2),Janus kinase 2(JAK2)and signal transduction and transcription activator 3(STAT3)were detected by RT-qPCR,Western blot was used to detect the protein expressions of Bax,Bcl-2,Caspace-3,Caspace-9,NDRG2,JAK2,STAT3,p-JAK2 and p-STAT3 in cells.Results Compared with the blank serum group,cell survival rate in methotrexate group,Quhan Zhufeng Mixture all dosage groups significantly decreased(P<0.01),the apoptosis rate significantly increased(P<0.01),the mRNA and protein expressions of Bax and Caspase-9 significantly increased(P<0.05,P<0.01),while the mRNA and protein expression of Bcl-2 significantly decreased(P<0.01),and Caspase-3 mRNA and protein expression in methotrexate group and Quhan Zhufeng Mixture medium-and high-dosage groups significantly increased(P<0.01).Compared with the blank serum group,the mRNA and protein expression of NDRG2 significantly increased in Quhan Zhufeng Mixture all dosage groups(P<0.05,P<0.01),the mRNA and protein expressions of JAK2 and STAT3 were significantly reduced in AG490 group and Quhan Zhufeng Mixture medium-and high-dosage groups(P<0.05,P<0.01),and the expressions of p-JAK2 and p-STAT3 proteins were significantly reduced(P<0.01).Conclusion Quhan Zhufeng Mixture can regulate the proliferation and apoptosis of RA-FLS by regulating the activity of NDRG2/JAK2/STAT3 signaling pathway,playing a role in treating rheumatoid arthritis.

Objective To explore the mechanism of Quhan Zhufeng Mixture on proliferation and apoptosis of rheumatoid arthritis fibroblast-like synoviocyte(RA-FLS)based on NDRG2/JAK2/STAT3 signaling pathway.Methods RA-FLS cells were cultured in vitro,and were divided into ① blank serum group,methotrexate group,Quhan Zhufeng Mixture low-,medium-and high-dosage groups;② blank serum group,AG490 group,Quhan Zhufeng Mixture low-,medium-and high-dosage groups.Different concentrations of drug-containing serum were used to intervene cells.Cell proliferation was detected by CCK-8 method,apoptosis was detected by flow cytometry,and mRNA expressions of Bax,Bcl-2,Caspace-3,Caspace-9,N-myc downstream regulatory gene 2(NDRG2),Janus kinase 2(JAK2)and signal transduction and transcription activator 3(STAT3)were detected by RT-qPCR,Western blot was used to detect the protein expressions of Bax,Bcl-2,Caspace-3,Caspace-9,NDRG2,JAK2,STAT3,p-JAK2 and p-STAT3 in cells.Results Compared with the blank serum group,cell survival rate in methotrexate group,Quhan Zhufeng Mixture all dosage groups significantly decreased(P<0.01),the apoptosis rate significantly increased(P<0.01),the mRNA and protein expressions of Bax and Caspase-9 significantly increased(P<0.05,P<0.01),while the mRNA and protein expression of Bcl-2 significantly decreased(P<0.01),and Caspase-3 mRNA and protein expression in methotrexate group and Quhan Zhufeng Mixture medium-and high-dosage groups significantly increased(P<0.01).Compared with the blank serum group,the mRNA and protein expression of NDRG2 significantly increased in Quhan Zhufeng Mixture all dosage groups(P<0.05,P<0.01),the mRNA and protein expressions of JAK2 and STAT3 were significantly reduced in AG490 group and Quhan Zhufeng Mixture medium-and high-dosage groups(P<0.05,P<0.01),and the expressions of p-JAK2 and p-STAT3 proteins were significantly reduced(P<0.01).Conclusion Quhan Zhufeng Mixture can regulate the proliferation and apoptosis of RA-FLS by regulating the activity of NDRG2/JAK2/STAT3 signaling pathway,playing a role in treating rheumatoid arthritis.

AIM:To investigate the mechanism through which aerobic exercise improves inflammatory senes-cence in SAMP8 mice via Janus kinase 3(JAK3)-signal transducer and activator of transcription 5α(STAT5α)signaling pathway and to provide novel insights for anti-inflammatory aging interventions.METHODS:Sixteen 28-week-old SAMP8 mice were randomly divided into model(Mod)group and exercise(Exe)group,with 8 mice in each group.Addi-tionally,8 senile SAMR1 mice served as control(Con)group.The mice in Exe group underwent an 8-week aerobic training regimen on a treadmill.We monitored the changes in hair quality and body weight,immune organ indexes,histomorpho-logical alterations in immune organs,the expression levels of spleen aging marker P16,and the serum and spleen levels of inflammatory factors[tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)]and aging indicator IL-2.Moreover,we examined the alterations in the mRNA and protein expression of IL-2,JAK3 and STAT5α in the spleen.RESULTS:(1)Compared with Con group,the mice in Mod group exhibited decreased hair luster and volume,with some instances of alopecia areata,increased body weight,markedly reduced spleen and thymus indexes,evident atrophy and senescence of immune organs,and markedly decreased concentration of IL-2 in the serum and spleen.The expression of P16 in the spleen,and the serum and spleen levels of inflammatory factors TNF-α and IL-1β were significantly increased.The mRNA expression of IL-2 and IL-2 receptor subunit gamma(IL-2RG)in the spleen was markedly decreased,whereas that of JAK3 and STAT5α was markedly increased.Furthermore,the protein expression of IL-2 in the spleen was markedly de-creased,whereas that of STAT5α and JAK3 was markedly increased.(2)Compared with Mod group,the mice in Exe group showed relatively vigorous hair growth and better hair luster,lower body weight,markedly increased spleen and thy-mus indexes,and delayed immune organ aging.The concentration of IL-2 in the serum and spleen was markedly in-creased,while the expression of P16 in the spleen was markedly decreased.The levels of TNF-α and IL-1β in the serum and spleen were markedly decreased.The mRNA expression of IL-2 and IL-2RG in the spleen was markedly increased,whereas that of JAK3 and STAT5α was markedly decreased.The protein levels of IL-2 were markedly increased,whereas those of STAT5α and JAK3 were markedly decreased.CONCLUSION:Aerobic exercise can delay inflammatory aging in mice possibly through the JAK3-STAT5α signaling pathway.

Objective: To evaluate the mutation rates of homologous recombination repair (HRR) genes and the impact of these mutations on the clinical characteristics of metastatic endometrial cancer (EC). Methods: Somatic DNA from 895 patients with metastatic EC in the Memorial Sloan Kettering-Metastatic Events and Tropisms cohort was assessed for mutations in 10 HRR genes (BRCA1, BRCA2, ATM, BARD1, BRIP1, PALB2, RAD51C, RAD51D, CHEK2, and CDK12).The correlation between the mutation status of HRR genes and the clinical characteristics of patients with metastatic EC was evaluated. Results: Somatic mutations in HRR genes were detected in 106 (11.8%) patients with metastatic EC. Compared with nonmutation carriers, a greater proportion of carriers had endometrioid carcinoma (76.4% vs. 50.3%, p<0.001). Regarding the TCGA classification, the proportions of the POLE-ultramutated (POLEmut) and mismatch repair-deficient (dMMR) subtypes were significantly greater among mutation carriers than noncarriers (20.8% vs.0.4%, p<0.001; 34.9% vs. 11.9%, p<0.001, respectively). The carriers had a significantly lower frequency of TP53 mutations than noncarriers (25.5% vs. 54.1%, p<0.001). Fewer mutation carriers than noncarriers had intra-abdominal and lung metastases (41.5% vs.54.2%, p=0.014; 19.8% vs. 30.3%, p=0.026, respectively). The mutation status of HRR genes did not significantly affect the overall survival of patients with metastatic EC. Conclusion Somatic HRR mutations are detected in 11.8% of metastatic EC. Compared with noncarriers, HRR mutation carriers in metastatic EC have higher proportions of endometrioid carcinoma, POLEmut, and dMMR subtypes, and unique metastatic patterns.However, the prognoses are similar regardless of HRR status.

Objective:To observe the effect of exercise-regulated miR-344g-5p on the fibrosis-related SMAD genes and transforming growth factor beta (TGF-β) in the myocardia of mice modeling diabetes.Methods:Twenty-four male C57BL/6 mice (6-8 weeks old) were randomly divided into a control group ( n=12) and a type 2 diabetes group ( n=12). Each group was further divided into two subgroups based on exercise status to form a sedentary control group, an exercise control, a sedentary type 2 diabetes group and an exercise type 2 diabetes group with six mice in each subgroup. The control groups were fed a normal diet, while the type 2 diabetes groups were on a high-fat diet for 12 weeks. Type 2 diabetes was then induced by intraperitoneal injection of streptozotocin. Two weeks later, the exercise groups began 40 minutes of daily swimming training, five days a week for eight consecutive weeks. Right after that, their cardiac function was measured using a small animal ultrasound system and the derived ejection fraction (EF) and the maximal early (E) and late (A) transmitral velocities ratio (E/A ratio) in diastole. They were then sacrificed and myocardial tissue was resected and stained with Sirius red. The expression of miR-344g-5p in the myocardium was detected using quantitative polymerase chain reactions. The expression of phosphorylated SMAD3 (p-SMAD3) and TGF-β were assessed using western blotting. The Target Scan database was exploited to analyze whether there were predicted targets of miR-344g-5p and pro-fibrotic genes such as SMAD3, TGFBR2, COL1A2 and COL12A1, and to determine any correlations in the gene regulation. Results:After 22 weeks, the EF and E/A ratio in the sedentary type 2 diabetes group were (57.5±4.1)% and (1.4±0.3), respectively, both significantly lower than in the other groups. Myocardial collagen fibers in the sedentary type 2 diabetes group were significantly more abundant than in the sedentary control and exercise type 2 diabetes groups. And miR-344g-5p expression in the myocardia of the exercise type 2 diabetes group was significantly greater than that in the sedentary type 2 diabetes group. The expression of p-SMAD3 and TGF-β in the myocardia of the sedentary type 2 diabetes group was significantly higher than in the sedentary control and exercise type 2 diabetes groups. Target Scan analysis revealed that miR-344g-5p had potential binding sites with several fibrosis-related genes such as SMAD3, TGFBR2, COL1A2, and COL12A1. Based on the reduction in TGF-β and p-SMAD protein expression in the exercise type 2 diabetes group, it was hypothesized that miR-344g-5p may inhibit the post-transcriptional processes of those genes.Conclusions:Exercise promotes the recovery of diabetic cardiomyopathy by upregulating myocardial miR-344g-5p expression, which subsequently targets and suppresses p-SMAD3 and TGF-β protein expression, thereby reducing diabetic myocardial fibrosis.