Irritable bowel syndrome （IBS） is a functional bowel disorder characterized primarily by abdominal pain and altered defecation habits. In recent years， traditional Chinese medicine （TCM） has made progress in multiple aspects of IBS research and treatment， including syndrome distribution， development of TCM formulas， clinical efficacy evaluation， external therapies， and psychosocial regulation. However， it still faces challenges such as over-reliance on symptomatic manifestations rather than biomarkers for diagnostic criteria， and the lack of high-quality evidence-based data supporting the efficacy of TCM formulas in treating IBS. This paper proposed that TCM diagnosis and treatment of IBS should adhere to the strategy of integrating the holistic concept with syndrome differentiation and treatment， combining TCM external therapies such as acupuncture， moxibustion and acupoint application）， and emphasizing individualized diagnosis and treatment for psychosomatic abnormalities. Future research should integrate multi-omics technologies， artificial intelligence and other methods to deepen the understanding of the pathogenesis of IBS and the mechanisms of TCM formulas， so as to promote the standardization and internationalization of TCM in the diagnosis and treatment of IBS.

Background and aims The mechanism of cholestatic liver injury (CLI) is unclear, and effective therapies are lacking. While peroxisome proliferator-activated receptor alpha (PPARα) agonists show potential hepatoprotective effect and pyroptosis is implicated in hepatocellular damage, how PPARα activation mitigates lithocholic acid (LCA)-induced pyroptosis remains unknown. Methods The hepatoprotective effect of PPARα agonists was evaluated in a mouse model of intrahepatic cholestasis induced by LCA. Liver injury was assessed via serum biochemistry, hematoxylin and eosin and TUNEL staining, and electron microscopy. Pyroptosis pathways were analyzed using real-time quantitative polymerase chain reaction, Western blot, and co-immunoprecipitation. Results Combined morphological, histopathological, and biochemical analyses confirmed that PPARα activation protects against CLI. Compared with LCA treatment alone, PPARα activation significantly attenuated the elevation of serum lactate dehydrogenase (LDH), the increased TUNEL-positive cells, and the formation of hepatocyte membrane pores. Mechanistically, PPARα activation suppressed both NOD-like receptor protein 3 (NLRP3) inflammasome-mediated pyroptosis and apoptosis protease-activating factor-1 (APAF-1)/CASPASE-3/GSDME-mediated pyroptosis. Furthermore, PPARα agonist pretreatment inhibited activation of the nuclear factor-kappa B (NF-κB) and forkhead box O1 (FOXO1) signaling pathways. Conclusions PPARα protects against LCA-induced CLI by inhibiting both NLRP3 inflammasome-mediated pyroptosis associated with NF-κB and APAF-1/CASPASE-3/GSDME-mediated pyroptosis associated with the FOXO1 signaling pathway.

Objective To investigate the effects and mechanism of Xuefu Zhuyu Decoction in oxidative stress in coronary heart disease model rats with heart blood stasis syndrome based on Keap1/Nrf2 signaling pathway.Methods The rats were divided into normal group,sham-operation group,model group,Xuefu Zhuyu Decoction group and trimetazidine group.The rat model of coronary heart disease with heart blood stasis syndrome was established by ligation of the left anterior descending branch of the coronary artery.Xuefu Zhuyu Decoction group and trimetazidine group were administrated with the corresponding drugs at the dosages of 14.04 g/kg and 5.4 mg/kg,respectively,and normal group,sham-operation group and model group were administrated with the same volume of normal saline for 14 days.The general state of rats was observed,body mass was recorded and electrocardiogram was collected.Echocardiography was used to examine cardiac functions(LVEF,LVFS,LVIDd,LVIDs);the morphology of myocardial tissue was observed by HE staining,serum malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and total antioxidant capacity(T-AOC)were detected by ELISA,the positive expressions of Keap1,Nrf2,HO-1 and NQO1 in myocardial tissue were detected by immunohistochemistry.Results Compared with the normal group and sham-operation group,the rats in the model group showed signs of mental fatigue,reduced activity,dull fur,purple claws,and a significant decrease in body mass(P<0.01);the ST segment in lead Ⅱ of the electrocardiogram was significantly elevated,LVEF and LVFS were significantly reduced,and LVIDd and LVIDs significantly increased(P<0.01),with severe degeneration and necrosis of myocardial cells,disappearance of striated structures,disordered arrangement of myocardial fibers,infiltration of inflammatory cells;the serum MDA content significantly increased,while the activities of SOD,GSH-Px and T-AOC significantly decreased(P<0.01);the positive expressions of Keap1 and Nrf2 in myocardial tissue significantly increased,while the positive expression of HO-1 and NQO1 significantly decreased(P<0.01).Compared with the model group,the rats in Xuefu Zhuyu Decoction group and trimetazidine group showed improvement in their mental state,increased activity,shiny fur,rosy nails,and significantly increased body mass(P<0.01);the ST segment of the electrocardiogram decreased to varying degrees,with significant increases in LVEF and LVFS,and significant decreases in LVIDd and LVIDs(P<0.01);a large number of myocardial cells survived,the arrangement of myocardial fibers was relatively regular,and the infiltration of inflammatory cells was significantly reduced;the serum MDA content was significantly reduced,while the activities of SOD,GSH-Px and T-AOC significantly increased(P<0.01);the positive expression of Keap1 in myocardial tissue significantly decreased,while the positive expressions of Nrf2,HO-1 and NQO1 significantly increased(P<0.01).Conclusion Xuefu Zhuyu Decoction may inhibit oxidative stress by activating Keap1/Nrf2 signaling pathway to improve the pathological morphology and structural damage of myocardial tissue and promote the recovery of cardiac functions in rats with heart blood stasis syndrome of coronary heart disease.

Aim To establish the pyroptosis model of rat chondrocytes induced by tumor necrosis factor α(TNF-α)in order to study the effect of lysine acetyl-transferase 7(KAT7)on pyroptosis of chondrocytes.Methods Chondrocytes of rat knee joint were isolated by type Ⅱ collagenase digestion,and were identified by toluidine blue staining and Col Ⅱ immunofluorescence.CCK-8 was used to evaluate cell viability.Western blot was used to detect the expression of pyroptosis-related proteins NLRP3,GSDMD,caspase-8 and KAT7 in cells intervened with TNF-α,adenovirus overexpression of KAT7(KAT7-oe)and KAT7 inhibitor WM-3835.The microstructure of the cells was observed by scanning e-lectron microscopy.Pyroptosis was detected by TUNEL staining,and the expression of pyroptosis-related pro-tein and KAT7 was detected by immunofluorescence.Results Compared with the empty virus group,KAT7-oe inhibited cell viability,promoted the expression of pyroptosis-related proteins,and TNF-α enhanced this effect.At the same time,the expression of KAT7 and pyroptosis-related proteins in the TNF-α stimulation group increased,and WM-3835 reduced the related proteins expression.Electron microscopy showed that KAT7-oe caused cell swelling,deformation,membrane perforation and rupture,while WM-3835 could restore cell morphology.TUNEL staining and immunofluores-cence results also confirmed that KAT7-oe induced chondrocyte pyroptosis,and WM-3835 could down-reg-ulate the fluorescence of pyroptosis-related proteins.Conclusions The expression of KAT7 increases in rat chondrocyte pyroptosis model,and the intervention of KAT7 expression affects signal molecules related to py-roptosis pathway,suggesting that KAT7 may be related to chondrocyte pyroptosis.

Objective To investigate the effects and mechanism of Xuefu Zhuyu Decoction in oxidative stress in coronary heart disease model rats with heart blood stasis syndrome based on Keap1/Nrf2 signaling pathway.Methods The rats were divided into normal group,sham-operation group,model group,Xuefu Zhuyu Decoction group and trimetazidine group.The rat model of coronary heart disease with heart blood stasis syndrome was established by ligation of the left anterior descending branch of the coronary artery.Xuefu Zhuyu Decoction group and trimetazidine group were administrated with the corresponding drugs at the dosages of 14.04 g/kg and 5.4 mg/kg,respectively,and normal group,sham-operation group and model group were administrated with the same volume of normal saline for 14 days.The general state of rats was observed,body mass was recorded and electrocardiogram was collected.Echocardiography was used to examine cardiac functions(LVEF,LVFS,LVIDd,LVIDs);the morphology of myocardial tissue was observed by HE staining,serum malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and total antioxidant capacity(T-AOC)were detected by ELISA,the positive expressions of Keap1,Nrf2,HO-1 and NQO1 in myocardial tissue were detected by immunohistochemistry.Results Compared with the normal group and sham-operation group,the rats in the model group showed signs of mental fatigue,reduced activity,dull fur,purple claws,and a significant decrease in body mass(P<0.01);the ST segment in lead Ⅱ of the electrocardiogram was significantly elevated,LVEF and LVFS were significantly reduced,and LVIDd and LVIDs significantly increased(P<0.01),with severe degeneration and necrosis of myocardial cells,disappearance of striated structures,disordered arrangement of myocardial fibers,infiltration of inflammatory cells;the serum MDA content significantly increased,while the activities of SOD,GSH-Px and T-AOC significantly decreased(P<0.01);the positive expressions of Keap1 and Nrf2 in myocardial tissue significantly increased,while the positive expression of HO-1 and NQO1 significantly decreased(P<0.01).Compared with the model group,the rats in Xuefu Zhuyu Decoction group and trimetazidine group showed improvement in their mental state,increased activity,shiny fur,rosy nails,and significantly increased body mass(P<0.01);the ST segment of the electrocardiogram decreased to varying degrees,with significant increases in LVEF and LVFS,and significant decreases in LVIDd and LVIDs(P<0.01);a large number of myocardial cells survived,the arrangement of myocardial fibers was relatively regular,and the infiltration of inflammatory cells was significantly reduced;the serum MDA content was significantly reduced,while the activities of SOD,GSH-Px and T-AOC significantly increased(P<0.01);the positive expression of Keap1 in myocardial tissue significantly decreased,while the positive expressions of Nrf2,HO-1 and NQO1 significantly increased(P<0.01).Conclusion Xuefu Zhuyu Decoction may inhibit oxidative stress by activating Keap1/Nrf2 signaling pathway to improve the pathological morphology and structural damage of myocardial tissue and promote the recovery of cardiac functions in rats with heart blood stasis syndrome of coronary heart disease.

Aim To establish the pyroptosis model of rat chondrocytes induced by tumor necrosis factor α(TNF-α)in order to study the effect of lysine acetyl-transferase 7(KAT7)on pyroptosis of chondrocytes.Methods Chondrocytes of rat knee joint were isolated by type Ⅱ collagenase digestion,and were identified by toluidine blue staining and Col Ⅱ immunofluorescence.CCK-8 was used to evaluate cell viability.Western blot was used to detect the expression of pyroptosis-related proteins NLRP3,GSDMD,caspase-8 and KAT7 in cells intervened with TNF-α,adenovirus overexpression of KAT7(KAT7-oe)and KAT7 inhibitor WM-3835.The microstructure of the cells was observed by scanning e-lectron microscopy.Pyroptosis was detected by TUNEL staining,and the expression of pyroptosis-related pro-tein and KAT7 was detected by immunofluorescence.Results Compared with the empty virus group,KAT7-oe inhibited cell viability,promoted the expression of pyroptosis-related proteins,and TNF-α enhanced this effect.At the same time,the expression of KAT7 and pyroptosis-related proteins in the TNF-α stimulation group increased,and WM-3835 reduced the related proteins expression.Electron microscopy showed that KAT7-oe caused cell swelling,deformation,membrane perforation and rupture,while WM-3835 could restore cell morphology.TUNEL staining and immunofluores-cence results also confirmed that KAT7-oe induced chondrocyte pyroptosis,and WM-3835 could down-reg-ulate the fluorescence of pyroptosis-related proteins.Conclusions The expression of KAT7 increases in rat chondrocyte pyroptosis model,and the intervention of KAT7 expression affects signal molecules related to py-roptosis pathway,suggesting that KAT7 may be related to chondrocyte pyroptosis.

Objective To investigate the independent risk factors for portal vein thrombosis(PVT)in patients with viral hepatitis-related decompensated cirrhosis,and to establish and validate a nomogram risk prediction model.Methods A retrospective analysis was performed for the clinical data of 1 116 patients with decompensated HBV/HCV cirrhosis who attended The Third People's Hospital of Kunming for the first time from January 2022 to December 2023,and according to the presence or absence of PVT,they were divided into PVT group and control group.The independent samples t-test or the Mann-Whitney U test was used for comparison of continuous data between groups,and the chi-square test was used for comparison of categorical data between groups.Univariate analysis and least absolute shrinkage and selection operator(LASSO)regression analysis were used to identify variables,and a binary logistic regression analysis was used to obtain independent influencing factors and establish a predictive model,which was visualized using a nomogram.The model was validated based on the receiver operating characteristic(ROC)curve,the area under the ROC curve(AUC),the Hosmer-Lemeshow test,Bootstrap sampling(1 000 iterations),the calibration curve,the decision curve analysis(DCA),and the clinical impact curve(CIC).Results There were 178 patients in the PVT group and 938 patients in the control group,and the prevalence rate of PVT was 15.9%(178/1 116).Male patients accounted for 68.5%(764/1 116),and the patients with drinking,Child-Pugh class B liver function,and ascites accounted for 51.0%(569/1 116),78.8%(879/1 116),and 67.1%(749/1 116),respectively.Compared with the control group,the PVT group had significantly higher age(Z=-2.362,P<0.05),prothrombin time(Z=-2.403,P<0.05),international normalized ratio(Z=-2.470,P<0.05),free thyroxine(Z=-5.910,P<0.05),D-dimer(Z=-5.764,P<0.05),interleukin-6(Z=-6.581,P<0.05),interleukin-10(IL-10)(Z=-3.915,P<0.05),interleukin-8(Z=-3.705,P<0.05),diameter of the portal vein(Z=-9.690,P<0.05),and spleen thickness(Z=-7.183,P<0.05),as well as significantly lower levels of white blood cell count(Z=-2.115,P<0.05),platelet count(Z=-3.026,P<0.05),fibrinogen(Z=-2.169,P<0.05),alanine aminotransferase(Z=-3.151,P<0.05),prealbumin(Z=-3.509,P<0.05),cholinesterase(Z=-3.415,P<0.05),alpha-fetoprotein(Z=-3.513,P<0.05),triglycerides(Z=-2.679,P<0.05),CD3 cell count(Z=-6.059,P<0.05),CD4 cell count(Z=-7.257,P<0.05),CD8 cell count(Z=-2.340,P<0.05),CD4+/CD8+cell ratio(Z=-4.479,P<0.05),triiodothyronine(Z=-3.338,P<0.05),free triiodothyronine(FT3)(Z=-9.560,P<0.05),and portal blood flow velocity(Z=-4.568,P<0.05).The multivariate logistic regression analysis was performed for the variables with statistical significance identified by the LASSO regression analysis,and the results showed that age(odds ratio[OR]=1.046,95%confidence interval[CI]:1.026-1.066),CD4+/CD8+cell ratio(OR=0.568,95%CI:0.410-0.787),FT3(OR=0.956,95%CI:0.944-0.968),IL-10(OR=1.021,95%CI:1.001-1.042),diameter of the portal vein(OR=1.446,95%CI:1.329-1.574),and spleen thickness(OR=1.035,95%CI:1.014-1.055)were independent influencing factors.A model was established as Logit(P)=-8.784+0.045×age-0.566×CD4+/CD8+-0.046×FT3+0.021×IL-10+0.369×diameter of the portal vein+0.034×spleen thickness,and a nomogram model was established and validated based on this model,with an AUC of 0.859(95%CI:0.833-0.887).The Hosmer-Lemeshow test showed that the model had a high goodness of fit(χ2=11.349,P=0.183).Bootstrap internal validation showed a mean absolute error of 0.006 and a C-index of 0.855.The decision curve analysis showed that the model had a high net clinical benefit within a wide range of thresholds.Conclusion Age,CD4+/CD8+ratio,FT3,IL-10,diameter of the portal vein,and spleen thickness may be independent influencing factors for PVT in patients with decompensated HBV/HCV cirrhosis.The predictive model established based on these six variables can help to predict the risk of PVT in patients with hepatitis-related decompensated cirrhosis in the early stage in clinical practice.

Objective To investigate the effects and mechanisms of replication activating factor C2(RFC2)on the proliferation and migration of renal cell carcinoma(RCC)cells.Methods Western blotting was used to determine and compare the expression levels of RFC2 protein in human normal renal tubular epithelial cell lines HK-2 and RCC cell lines ACHN,786-O,Caki-1,KETR3,and OS-RC2.The KETR-3 cell line was transfected with Lipofectamine 3 000 using RFC2 small interfering RNA(siRNA)and control inhibitory sequence scaffold,respectively,and divided into RFC2 silenced expression group(si-RFC2)and inhibitory control group(si-NC).Transfect RFC2 overexpression plasmid(RFC2 plasmid)and blank control plasmid,and divide into RFC2 overexpression group(OE-RFC2)and overexpression control group(Vector).Cell proliferation ability was detected using Methylthiazolyldiphenyl-tetrazolium bromide(MTT)cell proliferation detection kit,cell migration ability was measured using cell scratch assay,and protein blotting assay was used to determine the expression levels of RFC2,p-PI3 K,PI3 K,p-Akt,and Akt proteins.Results Compared with the normal human renal tubular epithelial cell line HK-2,the RCC cell lines ACHN,786-O,Caki-1,KETR3,and OS-RC2 showed high expression of RFC2 protein(all P＜0.001).MTT assay showed that the A490nm values of the si-RFC2 group were lower than those of the si-NC group at 0,24,48,and 72 h(all P＜0.05),while the A490nm values of the OE-RFC2 group were higher than those of the Vector group at 0,24,48,and 72 h(all P＜0.05).The scratch healing rate of the OE-RFC2 group was higher than that of the Vector group[(89.4±9.4)％vs(15.8±6.3)％,P＜0.05].The cell scratch healing rate in the si-RFC2 group was lower than that in the si-NC group[(5.2±1.9)％vs(16.5±5.5)％,P＜0.05].After upregulating RFC2 expression in KETR-3 cells,the relative expression level of RFC2 protein in the OE-RFC2 group was higher than that in the Vector group(1.04±0.19 vs 0.25±0.03,P＜0.05),the p-PI3K/PI3K ratio in the OE-RFC2 group was higher than that in the Vector group(1.15±0.23 vs 0.34±0.024,P＜0.05),and the p-Akt/Akt ratio in the OE-RFC2 group was higher than that in the Vector group(1.26±0.25 vs 0.38±0.022,P＜0.05).After silencing the expression of RFC2,the relative expression level of RFC2 protein in the si-RFC2 group was lower than that in the si-NC group(0.16±0.02 vs 1.08±0.06,P＜0.05),the p-PI3K/PI3K ratio in the si-RFC2 group was lower than that in the si-NC group(0.18±0.04 vs 0.86±0.14,P＜0.05),and the p-Akt/Akt ratio in the si-RFC2 group was lower than that in the si-NC group(0.10±0.01 vs0.58±0.13,P＜0.05).Conclusion RFC2 promotes the proliferation and migration of renal cell carcinoma cells,and the mechanism may be related to the activation of the PI3K/Akt signaling pathway.

Objective To investigate the independent risk factors for portal vein thrombosis(PVT)in patients with viral hepatitis-related decompensated cirrhosis,and to establish and validate a nomogram risk prediction model.Methods A retrospective analysis was performed for the clinical data of 1 116 patients with decompensated HBV/HCV cirrhosis who attended The Third People's Hospital of Kunming for the first time from January 2022 to December 2023,and according to the presence or absence of PVT,they were divided into PVT group and control group.The independent samples t-test or the Mann-Whitney U test was used for comparison of continuous data between groups,and the chi-square test was used for comparison of categorical data between groups.Univariate analysis and least absolute shrinkage and selection operator(LASSO)regression analysis were used to identify variables,and a binary logistic regression analysis was used to obtain independent influencing factors and establish a predictive model,which was visualized using a nomogram.The model was validated based on the receiver operating characteristic(ROC)curve,the area under the ROC curve(AUC),the Hosmer-Lemeshow test,Bootstrap sampling(1 000 iterations),the calibration curve,the decision curve analysis(DCA),and the clinical impact curve(CIC).Results There were 178 patients in the PVT group and 938 patients in the control group,and the prevalence rate of PVT was 15.9%(178/1 116).Male patients accounted for 68.5%(764/1 116),and the patients with drinking,Child-Pugh class B liver function,and ascites accounted for 51.0%(569/1 116),78.8%(879/1 116),and 67.1%(749/1 116),respectively.Compared with the control group,the PVT group had significantly higher age(Z=-2.362,P<0.05),prothrombin time(Z=-2.403,P<0.05),international normalized ratio(Z=-2.470,P<0.05),free thyroxine(Z=-5.910,P<0.05),D-dimer(Z=-5.764,P<0.05),interleukin-6(Z=-6.581,P<0.05),interleukin-10(IL-10)(Z=-3.915,P<0.05),interleukin-8(Z=-3.705,P<0.05),diameter of the portal vein(Z=-9.690,P<0.05),and spleen thickness(Z=-7.183,P<0.05),as well as significantly lower levels of white blood cell count(Z=-2.115,P<0.05),platelet count(Z=-3.026,P<0.05),fibrinogen(Z=-2.169,P<0.05),alanine aminotransferase(Z=-3.151,P<0.05),prealbumin(Z=-3.509,P<0.05),cholinesterase(Z=-3.415,P<0.05),alpha-fetoprotein(Z=-3.513,P<0.05),triglycerides(Z=-2.679,P<0.05),CD3 cell count(Z=-6.059,P<0.05),CD4 cell count(Z=-7.257,P<0.05),CD8 cell count(Z=-2.340,P<0.05),CD4+/CD8+cell ratio(Z=-4.479,P<0.05),triiodothyronine(Z=-3.338,P<0.05),free triiodothyronine(FT3)(Z=-9.560,P<0.05),and portal blood flow velocity(Z=-4.568,P<0.05).The multivariate logistic regression analysis was performed for the variables with statistical significance identified by the LASSO regression analysis,and the results showed that age(odds ratio[OR]=1.046,95%confidence interval[CI]:1.026-1.066),CD4+/CD8+cell ratio(OR=0.568,95%CI:0.410-0.787),FT3(OR=0.956,95%CI:0.944-0.968),IL-10(OR=1.021,95%CI:1.001-1.042),diameter of the portal vein(OR=1.446,95%CI:1.329-1.574),and spleen thickness(OR=1.035,95%CI:1.014-1.055)were independent influencing factors.A model was established as Logit(P)=-8.784+0.045×age-0.566×CD4+/CD8+-0.046×FT3+0.021×IL-10+0.369×diameter of the portal vein+0.034×spleen thickness,and a nomogram model was established and validated based on this model,with an AUC of 0.859(95%CI:0.833-0.887).The Hosmer-Lemeshow test showed that the model had a high goodness of fit(χ2=11.349,P=0.183).Bootstrap internal validation showed a mean absolute error of 0.006 and a C-index of 0.855.The decision curve analysis showed that the model had a high net clinical benefit within a wide range of thresholds.Conclusion Age,CD4+/CD8+ratio,FT3,IL-10,diameter of the portal vein,and spleen thickness may be independent influencing factors for PVT in patients with decompensated HBV/HCV cirrhosis.The predictive model established based on these six variables can help to predict the risk of PVT in patients with hepatitis-related decompensated cirrhosis in the early stage in clinical practice.

Objective To investigate the effects and mechanisms of replication activating factor C2(RFC2)on the proliferation and migration of renal cell carcinoma(RCC)cells.Methods Western blotting was used to determine and compare the expression levels of RFC2 protein in human normal renal tubular epithelial cell lines HK-2 and RCC cell lines ACHN,786-O,Caki-1,KETR3,and OS-RC2.The KETR-3 cell line was transfected with Lipofectamine 3 000 using RFC2 small interfering RNA(siRNA)and control inhibitory sequence scaffold,respectively,and divided into RFC2 silenced expression group(si-RFC2)and inhibitory control group(si-NC).Transfect RFC2 overexpression plasmid(RFC2 plasmid)and blank control plasmid,and divide into RFC2 overexpression group(OE-RFC2)and overexpression control group(Vector).Cell proliferation ability was detected using Methylthiazolyldiphenyl-tetrazolium bromide(MTT)cell proliferation detection kit,cell migration ability was measured using cell scratch assay,and protein blotting assay was used to determine the expression levels of RFC2,p-PI3 K,PI3 K,p-Akt,and Akt proteins.Results Compared with the normal human renal tubular epithelial cell line HK-2,the RCC cell lines ACHN,786-O,Caki-1,KETR3,and OS-RC2 showed high expression of RFC2 protein(all P＜0.001).MTT assay showed that the A490nm values of the si-RFC2 group were lower than those of the si-NC group at 0,24,48,and 72 h(all P＜0.05),while the A490nm values of the OE-RFC2 group were higher than those of the Vector group at 0,24,48,and 72 h(all P＜0.05).The scratch healing rate of the OE-RFC2 group was higher than that of the Vector group[(89.4±9.4)％vs(15.8±6.3)％,P＜0.05].The cell scratch healing rate in the si-RFC2 group was lower than that in the si-NC group[(5.2±1.9)％vs(16.5±5.5)％,P＜0.05].After upregulating RFC2 expression in KETR-3 cells,the relative expression level of RFC2 protein in the OE-RFC2 group was higher than that in the Vector group(1.04±0.19 vs 0.25±0.03,P＜0.05),the p-PI3K/PI3K ratio in the OE-RFC2 group was higher than that in the Vector group(1.15±0.23 vs 0.34±0.024,P＜0.05),and the p-Akt/Akt ratio in the OE-RFC2 group was higher than that in the Vector group(1.26±0.25 vs 0.38±0.022,P＜0.05).After silencing the expression of RFC2,the relative expression level of RFC2 protein in the si-RFC2 group was lower than that in the si-NC group(0.16±0.02 vs 1.08±0.06,P＜0.05),the p-PI3K/PI3K ratio in the si-RFC2 group was lower than that in the si-NC group(0.18±0.04 vs 0.86±0.14,P＜0.05),and the p-Akt/Akt ratio in the si-RFC2 group was lower than that in the si-NC group(0.10±0.01 vs0.58±0.13,P＜0.05).Conclusion RFC2 promotes the proliferation and migration of renal cell carcinoma cells,and the mechanism may be related to the activation of the PI3K/Akt signaling pathway.