Objective:To explore the mechanism of astaxanthin improving renal damage in diabetic mice by regulating the PI3K/Akt signaling pathway.Methods:C57BL/6J adult male mice (8 weeks, 22-24 g) were provided by Nanjing Junke Biological Co.,Ltd. The mice were divided into control group (mice raised under normal conditions and given phosphate buffered saline injection, n=15), model group (DN mouse model established as mentioned above, n=15), and astaxanthin group (on the basis of model mice,10 mg/kg body weight dose of astaxanthin was given, n=15). The serum urea nitrogen, serum creatinine and 24 h urine protein levels of mice were detected by biochemical kits. The levels of serum inflammatory factors in mice were detected by ELISA. Mesangial matrix expansion and fibrosis in mice were observed by renal histological analysis. Glomerular podocytes were analyzed by TUNEL detection and immunohistochemical staining. Nephrin and CD2AP expression were analyzed by Western blot.The expression of PI3K/Akt signaling pathway was analyzed by Western blot. Results:The levels of serum urea nitrogen, serum creatinine and 24h urinary protein in model group were higher than those in control group ( P<0.05), but the levels of serum urea nitrogen, serum creatinine and 24h urinary protein in astaxanthin group were lower than those in model group ( P<0.05). The serum levels of TNF-α,1L-1β and 1L-6 in model group were higher than those in control group ( P<0.05), while the levels of TNF-α,1L-1β and 1L-6 in astaxanthin group were lower than those in model group ( P<0.05). Compared with the control group, the model group mainly showed different degrees of pancreatic islet lesions and vacuolar degeneration under light microscope ( P<0.05). HE staining showed glomerular sclerosis and dilatation, capillary lumen shrinkage, diffuse mesangial matrix dilatation, and peripheral capillary thickening and hardening ( P<0.05). PAS staining showed an increase in PAS-positive substances (purple plaques) in the model group of mice ( P<0.05), indicating glycogen accumulation in diabetic glomeruli. Masson staining showed accumulation of type Ⅳ collagen and increased fibrosis (blue stained area) in the kidney of the model group ( P<0.05). Astaxanthin treatment can significantly improve these diabetic induced histopathological changes ( P<0.05). Compared with control group,mesangial matrix expansion and fibrosis were increased in model group ( P<0.05), and decreased in astaxanthin group ( P<0.05). Compared with the control group, the apoptosis rate of podocyte in model group was increased ( P<0.05) ,while that in astaxanthin group was decreased ( P<0.05). The number of WT-1 positive podocytes in model group was lower than that in model group ( P<0.05), and the number of WT-1 positive podocytes in astaxanthin group was higher than that in model group ( P<0.05). Compared with the control group, Nephrin and CD2AP proteins in the model group were decreased ( P<0.05), and the expressions of Nephrin and CD2AP proteins in astaxanthin group were increased ( P<0.05). The protein expressions of p85, p-Akt Ser473 and P-mtor Ser2448 in model group were increased compared with those in control group ( P<0.05), while the protein expressions of p85, P-Akt Ser473 and P-mtor Ser2448 in astaxanthin group were decreased compared with those in model group ( P<0.05) . Conclusion:Astaxanthin significantly improves kidney damage in diabetic mice by regulating the PI3K/Akt signaling pathway,which manifests as inhibiting renal cell lesions and reducing inflammation.

Objective:To explore the mechanism of astaxanthin improving renal damage in diabetic mice by regulating the PI3K/Akt signaling pathway.Methods:C57BL/6J adult male mice (8 weeks, 22-24 g) were provided by Nanjing Junke Biological Co.,Ltd. The mice were divided into control group (mice raised under normal conditions and given phosphate buffered saline injection, n=15), model group (DN mouse model established as mentioned above, n=15), and astaxanthin group (on the basis of model mice,10 mg/kg body weight dose of astaxanthin was given, n=15). The serum urea nitrogen, serum creatinine and 24 h urine protein levels of mice were detected by biochemical kits. The levels of serum inflammatory factors in mice were detected by ELISA. Mesangial matrix expansion and fibrosis in mice were observed by renal histological analysis. Glomerular podocytes were analyzed by TUNEL detection and immunohistochemical staining. Nephrin and CD2AP expression were analyzed by Western blot.The expression of PI3K/Akt signaling pathway was analyzed by Western blot. Results:The levels of serum urea nitrogen, serum creatinine and 24h urinary protein in model group were higher than those in control group ( P<0.05), but the levels of serum urea nitrogen, serum creatinine and 24h urinary protein in astaxanthin group were lower than those in model group ( P<0.05). The serum levels of TNF-α,1L-1β and 1L-6 in model group were higher than those in control group ( P<0.05), while the levels of TNF-α,1L-1β and 1L-6 in astaxanthin group were lower than those in model group ( P<0.05). Compared with the control group, the model group mainly showed different degrees of pancreatic islet lesions and vacuolar degeneration under light microscope ( P<0.05). HE staining showed glomerular sclerosis and dilatation, capillary lumen shrinkage, diffuse mesangial matrix dilatation, and peripheral capillary thickening and hardening ( P<0.05). PAS staining showed an increase in PAS-positive substances (purple plaques) in the model group of mice ( P<0.05), indicating glycogen accumulation in diabetic glomeruli. Masson staining showed accumulation of type Ⅳ collagen and increased fibrosis (blue stained area) in the kidney of the model group ( P<0.05). Astaxanthin treatment can significantly improve these diabetic induced histopathological changes ( P<0.05). Compared with control group,mesangial matrix expansion and fibrosis were increased in model group ( P<0.05), and decreased in astaxanthin group ( P<0.05). Compared with the control group, the apoptosis rate of podocyte in model group was increased ( P<0.05) ,while that in astaxanthin group was decreased ( P<0.05). The number of WT-1 positive podocytes in model group was lower than that in model group ( P<0.05), and the number of WT-1 positive podocytes in astaxanthin group was higher than that in model group ( P<0.05). Compared with the control group, Nephrin and CD2AP proteins in the model group were decreased ( P<0.05), and the expressions of Nephrin and CD2AP proteins in astaxanthin group were increased ( P<0.05). The protein expressions of p85, p-Akt Ser473 and P-mtor Ser2448 in model group were increased compared with those in control group ( P<0.05), while the protein expressions of p85, P-Akt Ser473 and P-mtor Ser2448 in astaxanthin group were decreased compared with those in model group ( P<0.05) . Conclusion:Astaxanthin significantly improves kidney damage in diabetic mice by regulating the PI3K/Akt signaling pathway,which manifests as inhibiting renal cell lesions and reducing inflammation.

Objective To investigate the relationship between serum arginase-1(Arg-1),thrombomodulin motif 9(ADAMTS-9),Ectodysplasin A and cardiac function after percutaneous coronary intervention(PCI)in patients with acute myocardial infarction(AMI),and their predictive value for major adverse cardiovascular events(MACE).Methods A total of 102 AMI patients who underwent PCI in the Third Medical Center of People's Liberation Army General Hospital from January 2021 to December 2023 were selected as the AMI group,and 110 healthy people were recruited as the control group.The serum levels of Arg-1,ADAMTS-9 and Ectodysplasin A were detected by enzyme-linked immunosorbent assay.Echocardiography was performed by color Doppler ultrasound,and left ventricular mass index(LVMI)was calculated.Patients were divided into MACE group(n=32)and non-MACE group(n=70)according to the occurrence of MACE.Pearson correla-tion analysis was used to analyze the correlation between serum Arg-1,ADAMTS-9,Ectodysplasin A levels and LVMI after PCI in AMI patients.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of serum Arg-1,ADAMTS-9 and Ectodysplasin A levels for MACE in AMI patients after PCI.Z test was used to compare the area under the curve(AUC).Multivariate Logistic regression analysis was used to analyze the influencing factors of MACE in patients with AMI.Results Compared with the con-trol group,the AMI group had significantly higher serum levels of Arg-1,ADAMTS-9 and Ectodysplasin A(P<0.05).The serum levels of Arg-1,ADAMTS-9 and Ectodysplasin A in patients with AMI were positively correlated with LVMI(P<0.05).There were no significant differences in gender,body mass index,systolic blood pressure,heart rate,history of diabetes,diastolic blood pressure,smoking history,time from onset to treatment,drinking history,age,course of disease,and LVMI between MACE group and non-MACE group(P>0.05).The serum levels of Arg-1,ADAMTS-9 and Ectodysplasin A in the MACE group were higher than those in the non-MACE group(P<0.05).Arg-1,ADAMTS-9 and Ectodysplasin A were independent risk factors for MACE in AMI patients(P<0.05).The AUC of serum Arg-1,ADAMTS-9 and Ectodysplasin A levels in predicting MACE in AMI patients were 0.821,0.779,0.818 and 0.950,respectively.The combined prediction value of the three was higher than that of individual prediction(Z=3.137,3.702,2.699,P=0.002,<0.001,0.007).Conclusion The serum levels of Arg-1,ADAMTS-9 and Ectodysplasin A are increased in AMI patients,and they have certain predictive value for the occurrence of MACE in AMI patients after PCI.

Objective To build the early predictive model for chronic kidney disease(CKD)in hypertension and diabetes patients in the community.Methods The CKD patients were recruited from 4 health care centers in 4 urban areas in Kunming.The control group was residents without hypertension and diabetes(n = 1267).The disease group was residents with hypertension and/or diabetes(n = 566).The questionnaire survey,physical examination,laboratory testing,and 5 SNPs gene types in the PVT1 gene.The risk factors,which were filtered with logistics regression,were used to build predictive models.Four machine learning algorithms were built:support vector machine(SVM),random forest(RF),Na?ve Bayes(NB),and artificial neural network(ANN)models.Results Thirteen indicators included in the final diagnostic model:age,disease type,ethnicity,blood urea nitrogen,creatinine,eGFR from MDRD,ACR,eGFR from EPI2009,PAM13 score,sleep quality survey,staying-up late,PVT1 SNP rs11993333 and rs2720659.The accuracy,specificity,Kappa value,AUC of ROC,and PRC of ANN are greater than those of the other 3 models.The sensitivity of RF is the highest among 4 types of machine learning.Conclusions The ANN predictive model has a good ability of efficiency and classification to predict CKD with hypertension and/or diabetes patients in the community.

Objective:To analyze the relationship between Twist2 and the prognosis and vascular infiltration of ovarian cancer patients,and to explore the role and mechanism of Twist2 in vascular infiltration of ovarian cancer.Methods:KM plotter was used to explore the correaltion between Twist2 mRNA expression and overall survival(OS),progression free survival(PFS)and post progression survival(PPS)in ovarian cancer.To analyze the cor-relation between Twist2 and vascular infiltration in ovarian cancer using the cancer genomics database cBioPor-tal.In vitro experiment:Transwell method was employed to determine the role of Twist2 in the invasion and migra-tion abilities of ovarian cancer cell CAOV3[blank group,negative control group(siNC group),siTwist2 group].Uti-lizing Realtime-PCR and Western blot to clarify the changes in Twist2 and VEGFC expression in CAOV3 cells af-ter downregulating Twist2 expression at the RNA and protein levels,respectively.Results:①Online data analysis of KM plotter showed that the ovarian cancer patients with high expression of Twist2 were associated with poor prognosis,with OS(HR 1.24,95％Cl 1.01-1.52),PFS(HR 1.39,95％CI 1.14-1.70)and PPS(HR 1.37,95％CI 1.08-1.74)all showing statistical significance(P＜0.05).CBioportal analysis showed that Twist2 mRNA expression was positively correlated with vascular infiltration(r=0.93,P=0.001)and lymphatic infiltration(r=0.89,P=0.009)in ovarian cancer.②Compared with the blank group and siNC group,in vitro experiment Tran-swell assay showed that the invasion and migration ability of ovarian cancer cells in siTwist2 group was significant-ly reduced(P＜0.05).In mRNA and protein level,Realtime-PCR and Western blot showed that compared with the blank group and siNC group,the expression of Twist2,as well as VEGFC,were significantly reduced in the siTwist2 group(P＜0.05).Conclusions:The expression of Twist2 in ovarian cancer is closely related to tumor prognosis and vascular infiltration.After downregulating Twist2,the number of cells that migrate and invade is sig-nificantly reduced,and the expression of VEGFC is reduced.Twist2 can induce migration and invasion of ovarian cancer cells through VEGFC,which may be one of the indicators for prognosis evaluate and targeted therapy of ovarian cancer in future.

Objective:To analyze the relationship between Twist2 and the prognosis and vascular infiltration of ovarian cancer patients,and to explore the role and mechanism of Twist2 in vascular infiltration of ovarian cancer.Methods:KM plotter was used to explore the correaltion between Twist2 mRNA expression and overall survival(OS),progression free survival(PFS)and post progression survival(PPS)in ovarian cancer.To analyze the cor-relation between Twist2 and vascular infiltration in ovarian cancer using the cancer genomics database cBioPor-tal.In vitro experiment:Transwell method was employed to determine the role of Twist2 in the invasion and migra-tion abilities of ovarian cancer cell CAOV3[blank group,negative control group(siNC group),siTwist2 group].Uti-lizing Realtime-PCR and Western blot to clarify the changes in Twist2 and VEGFC expression in CAOV3 cells af-ter downregulating Twist2 expression at the RNA and protein levels,respectively.Results:①Online data analysis of KM plotter showed that the ovarian cancer patients with high expression of Twist2 were associated with poor prognosis,with OS(HR 1.24,95％Cl 1.01-1.52),PFS(HR 1.39,95％CI 1.14-1.70)and PPS(HR 1.37,95％CI 1.08-1.74)all showing statistical significance(P＜0.05).CBioportal analysis showed that Twist2 mRNA expression was positively correlated with vascular infiltration(r=0.93,P=0.001)and lymphatic infiltration(r=0.89,P=0.009)in ovarian cancer.②Compared with the blank group and siNC group,in vitro experiment Tran-swell assay showed that the invasion and migration ability of ovarian cancer cells in siTwist2 group was significant-ly reduced(P＜0.05).In mRNA and protein level,Realtime-PCR and Western blot showed that compared with the blank group and siNC group,the expression of Twist2,as well as VEGFC,were significantly reduced in the siTwist2 group(P＜0.05).Conclusions:The expression of Twist2 in ovarian cancer is closely related to tumor prognosis and vascular infiltration.After downregulating Twist2,the number of cells that migrate and invade is sig-nificantly reduced,and the expression of VEGFC is reduced.Twist2 can induce migration and invasion of ovarian cancer cells through VEGFC,which may be one of the indicators for prognosis evaluate and targeted therapy of ovarian cancer in future.

Objective:To analyze the relationship between Twist2 and the prognosis and vascular infiltration of ovarian cancer patients,and to explore the role and mechanism of Twist2 in vascular infiltration of ovarian cancer.Methods:KM plotter was used to explore the correaltion between Twist2 mRNA expression and overall survival(OS),progression free survival(PFS)and post progression survival(PPS)in ovarian cancer.To analyze the cor-relation between Twist2 and vascular infiltration in ovarian cancer using the cancer genomics database cBioPor-tal.In vitro experiment:Transwell method was employed to determine the role of Twist2 in the invasion and migra-tion abilities of ovarian cancer cell CAOV3[blank group,negative control group(siNC group),siTwist2 group].Uti-lizing Realtime-PCR and Western blot to clarify the changes in Twist2 and VEGFC expression in CAOV3 cells af-ter downregulating Twist2 expression at the RNA and protein levels,respectively.Results:①Online data analysis of KM plotter showed that the ovarian cancer patients with high expression of Twist2 were associated with poor prognosis,with OS(HR 1.24,95％Cl 1.01-1.52),PFS(HR 1.39,95％CI 1.14-1.70)and PPS(HR 1.37,95％CI 1.08-1.74)all showing statistical significance(P＜0.05).CBioportal analysis showed that Twist2 mRNA expression was positively correlated with vascular infiltration(r=0.93,P=0.001)and lymphatic infiltration(r=0.89,P=0.009)in ovarian cancer.②Compared with the blank group and siNC group,in vitro experiment Tran-swell assay showed that the invasion and migration ability of ovarian cancer cells in siTwist2 group was significant-ly reduced(P＜0.05).In mRNA and protein level,Realtime-PCR and Western blot showed that compared with the blank group and siNC group,the expression of Twist2,as well as VEGFC,were significantly reduced in the siTwist2 group(P＜0.05).Conclusions:The expression of Twist2 in ovarian cancer is closely related to tumor prognosis and vascular infiltration.After downregulating Twist2,the number of cells that migrate and invade is sig-nificantly reduced,and the expression of VEGFC is reduced.Twist2 can induce migration and invasion of ovarian cancer cells through VEGFC,which may be one of the indicators for prognosis evaluate and targeted therapy of ovarian cancer in future.

Objective:To analyze the relationship between Twist2 and the prognosis and vascular infiltration of ovarian cancer patients,and to explore the role and mechanism of Twist2 in vascular infiltration of ovarian cancer.Methods:KM plotter was used to explore the correaltion between Twist2 mRNA expression and overall survival(OS),progression free survival(PFS)and post progression survival(PPS)in ovarian cancer.To analyze the cor-relation between Twist2 and vascular infiltration in ovarian cancer using the cancer genomics database cBioPor-tal.In vitro experiment:Transwell method was employed to determine the role of Twist2 in the invasion and migra-tion abilities of ovarian cancer cell CAOV3[blank group,negative control group(siNC group),siTwist2 group].Uti-lizing Realtime-PCR and Western blot to clarify the changes in Twist2 and VEGFC expression in CAOV3 cells af-ter downregulating Twist2 expression at the RNA and protein levels,respectively.Results:①Online data analysis of KM plotter showed that the ovarian cancer patients with high expression of Twist2 were associated with poor prognosis,with OS(HR 1.24,95％Cl 1.01-1.52),PFS(HR 1.39,95％CI 1.14-1.70)and PPS(HR 1.37,95％CI 1.08-1.74)all showing statistical significance(P＜0.05).CBioportal analysis showed that Twist2 mRNA expression was positively correlated with vascular infiltration(r=0.93,P=0.001)and lymphatic infiltration(r=0.89,P=0.009)in ovarian cancer.②Compared with the blank group and siNC group,in vitro experiment Tran-swell assay showed that the invasion and migration ability of ovarian cancer cells in siTwist2 group was significant-ly reduced(P＜0.05).In mRNA and protein level,Realtime-PCR and Western blot showed that compared with the blank group and siNC group,the expression of Twist2,as well as VEGFC,were significantly reduced in the siTwist2 group(P＜0.05).Conclusions:The expression of Twist2 in ovarian cancer is closely related to tumor prognosis and vascular infiltration.After downregulating Twist2,the number of cells that migrate and invade is sig-nificantly reduced,and the expression of VEGFC is reduced.Twist2 can induce migration and invasion of ovarian cancer cells through VEGFC,which may be one of the indicators for prognosis evaluate and targeted therapy of ovarian cancer in future.

Objective:To analyze the relationship between Twist2 and the prognosis and vascular infiltration of ovarian cancer patients,and to explore the role and mechanism of Twist2 in vascular infiltration of ovarian cancer.Methods:KM plotter was used to explore the correaltion between Twist2 mRNA expression and overall survival(OS),progression free survival(PFS)and post progression survival(PPS)in ovarian cancer.To analyze the cor-relation between Twist2 and vascular infiltration in ovarian cancer using the cancer genomics database cBioPor-tal.In vitro experiment:Transwell method was employed to determine the role of Twist2 in the invasion and migra-tion abilities of ovarian cancer cell CAOV3[blank group,negative control group(siNC group),siTwist2 group].Uti-lizing Realtime-PCR and Western blot to clarify the changes in Twist2 and VEGFC expression in CAOV3 cells af-ter downregulating Twist2 expression at the RNA and protein levels,respectively.Results:①Online data analysis of KM plotter showed that the ovarian cancer patients with high expression of Twist2 were associated with poor prognosis,with OS(HR 1.24,95％Cl 1.01-1.52),PFS(HR 1.39,95％CI 1.14-1.70)and PPS(HR 1.37,95％CI 1.08-1.74)all showing statistical significance(P＜0.05).CBioportal analysis showed that Twist2 mRNA expression was positively correlated with vascular infiltration(r=0.93,P=0.001)and lymphatic infiltration(r=0.89,P=0.009)in ovarian cancer.②Compared with the blank group and siNC group,in vitro experiment Tran-swell assay showed that the invasion and migration ability of ovarian cancer cells in siTwist2 group was significant-ly reduced(P＜0.05).In mRNA and protein level,Realtime-PCR and Western blot showed that compared with the blank group and siNC group,the expression of Twist2,as well as VEGFC,were significantly reduced in the siTwist2 group(P＜0.05).Conclusions:The expression of Twist2 in ovarian cancer is closely related to tumor prognosis and vascular infiltration.After downregulating Twist2,the number of cells that migrate and invade is sig-nificantly reduced,and the expression of VEGFC is reduced.Twist2 can induce migration and invasion of ovarian cancer cells through VEGFC,which may be one of the indicators for prognosis evaluate and targeted therapy of ovarian cancer in future.

Objective:To analyze the relationship between Twist2 and the prognosis and vascular infiltration of ovarian cancer patients,and to explore the role and mechanism of Twist2 in vascular infiltration of ovarian cancer.Methods:KM plotter was used to explore the correaltion between Twist2 mRNA expression and overall survival(OS),progression free survival(PFS)and post progression survival(PPS)in ovarian cancer.To analyze the cor-relation between Twist2 and vascular infiltration in ovarian cancer using the cancer genomics database cBioPor-tal.In vitro experiment:Transwell method was employed to determine the role of Twist2 in the invasion and migra-tion abilities of ovarian cancer cell CAOV3[blank group,negative control group(siNC group),siTwist2 group].Uti-lizing Realtime-PCR and Western blot to clarify the changes in Twist2 and VEGFC expression in CAOV3 cells af-ter downregulating Twist2 expression at the RNA and protein levels,respectively.Results:①Online data analysis of KM plotter showed that the ovarian cancer patients with high expression of Twist2 were associated with poor prognosis,with OS(HR 1.24,95％Cl 1.01-1.52),PFS(HR 1.39,95％CI 1.14-1.70)and PPS(HR 1.37,95％CI 1.08-1.74)all showing statistical significance(P＜0.05).CBioportal analysis showed that Twist2 mRNA expression was positively correlated with vascular infiltration(r=0.93,P=0.001)and lymphatic infiltration(r=0.89,P=0.009)in ovarian cancer.②Compared with the blank group and siNC group,in vitro experiment Tran-swell assay showed that the invasion and migration ability of ovarian cancer cells in siTwist2 group was significant-ly reduced(P＜0.05).In mRNA and protein level,Realtime-PCR and Western blot showed that compared with the blank group and siNC group,the expression of Twist2,as well as VEGFC,were significantly reduced in the siTwist2 group(P＜0.05).Conclusions:The expression of Twist2 in ovarian cancer is closely related to tumor prognosis and vascular infiltration.After downregulating Twist2,the number of cells that migrate and invade is sig-nificantly reduced,and the expression of VEGFC is reduced.Twist2 can induce migration and invasion of ovarian cancer cells through VEGFC,which may be one of the indicators for prognosis evaluate and targeted therapy of ovarian cancer in future.