Objective:To investigate the effect of lanatoside C on autophagy in hepatocellular carcinoma Huh-7 cells,and the possible mechanism of action.Methods:After Huh-7 cells were treated with gradient concentration of lanatoside C for 24 h,the IC50 value of lanatoside C on Huh-7 cells was detected by CCK-8 assay,and the role of intervention of gradient concentration of lanatoside C in the proliferation and clone formation of Huh-7 cells was analyzed by CCK-8 assay and clone formation assay.Western blotting was used to detect the expression levels of autophagy markers in Huh-7 cells,such as microtubule-associated protein light chain 3(LC3)-II,LC3-Ⅰ,p62,Beclin1,vacuolar protein sorting 34(VPS30),VPS15 and autophagy-related protein 3(ATG3)in Huh-7 cells,and the changes of LC3-II/LC3-Ⅰ ratio were analyzed.The effects of lanatoside C on the formation of autophagic vesicles in Huh-7 cells were observed by laser confocal microscopy,and the effects of lanatoside C on the formation of autophagic vesicles in Huh-7 cells were confirmed by artificial intelligence image analysis.Finally,the effects of lanatoside C on the interaction between Beclin1,VPS34 and VPS15 on the formation of autophagic vesicles in Huh-7 cells were detected by co-immunoprecipitation.Results:The IC50 value of lanatoside C on Huh-7 cells at 24 h was(0.49±0.07)μg/mL.The results of the CCK-8 assay and clone formation assay suggested that lanatoside C had a significant inhibitory effect on the growth of Huh-7 cells in a concentration-dependent manner(both P<0.01).The results of Western blotting assay showed that the LC3-II/LC3-Ⅰ ratio was significantly increased with the increase in the concentration of lanatoside C(P<0.01),while the expression level of Beclin1 protein was up-regulated(P<0.05),and the expression level of p62 protein was significantly down-regulated(P<0.01).The results of laser confocal microscopy showed that lanatoside C could significantly induce autophagic vesicle formation in Huh-7 cells(P<0.01).The results of artificial intelligence image analysis confirmed this effect of lanatoside C(all P<0.01).The results of co-immunoprecipitation confirmed that lanatoside C was able to promote the binding of Beclin1 to VPS34 and VPS15 with each other(both P<0.01).Conclusion:Lanatoside C significantly inhibits the proliferation of Huh-7 cells and and induces Huh-7 cells autophagy,the mechanism of this effect may be related to up-regulating Beclin1 expression and promoting the combination of Beclin1 protein with VPS34 and VPS15 proteins.

Objective:To explore the role of mechanical hemodynamic support (MHS) in mapping and catheter ablation of patients with hemodynamically unstable ventricular tachycardia (VT), report single-center experience in a cohort of consecutive patients receiving VT ablation during MHS therapy, and provide evidence-based medical evidence for clinical practice.Methods:This was a retrospective cohort study. Patients with hemodynamically unstable VT who underwent catheter ablation with MHS at Beijing Anzhen Hospital, Capital Medical University between August 2021 and December 2023 were included. Patients were divided into rescue group and preventive group according to the purpose of treatment. Their demographic data, periprocedural details, and clinical outcomes were collected and analyzed.Results:A total of 15 patients with hemodynamically unstable VT were included (8 patients in the rescue group and 7 patients in the preventive group). The acute procedure was successful in all patients. One patient in the rescue group had surgical left ventricular assist device (LVAD) implantation, remaining 14 patients received extracorporeal membrane oxygenation (ECMO) for circulation support. ECMO decannulation was performed in 12 patients due to clinical and hemodynamic stability, of which 6 patients were decannulation immediately after surgery and the remaining patients were decannulation at 2.0 (2.5) d after surgery. Two patients in the rescue group died during the index admission due to refractory heart failure and cerebral hemorrhage. During a median follow-up of 30 d (1 d to 12 months), one patient with LVAD had one episode of ventricular fibrillation at 6 months after discharge, and no further episodes of ventricular fibrillation and/or VT occurred after treatment with antiarrhythmic drugs. No malignant ventricular arrhythmia occurred in the remaining 12 patients who were followed up.Conclusions:MHS contributes to the successful completion of mapping and catheter ablation in patients with hemodynamically unstable VT, providing desirable hemodynamic status for emergency and elective conditions.

Objective:To investigate the effect of lanatoside C on autophagy in hepatocellular carcinoma Huh-7 cells,and the possible mechanism of action.Methods:After Huh-7 cells were treated with gradient concentration of lanatoside C for 24 h,the IC50 value of lanatoside C on Huh-7 cells was detected by CCK-8 assay,and the role of intervention of gradient concentration of lanatoside C in the proliferation and clone formation of Huh-7 cells was analyzed by CCK-8 assay and clone formation assay.Western blotting was used to detect the expression levels of autophagy markers in Huh-7 cells,such as microtubule-associated protein light chain 3(LC3)-II,LC3-Ⅰ,p62,Beclin1,vacuolar protein sorting 34(VPS30),VPS15 and autophagy-related protein 3(ATG3)in Huh-7 cells,and the changes of LC3-II/LC3-Ⅰ ratio were analyzed.The effects of lanatoside C on the formation of autophagic vesicles in Huh-7 cells were observed by laser confocal microscopy,and the effects of lanatoside C on the formation of autophagic vesicles in Huh-7 cells were confirmed by artificial intelligence image analysis.Finally,the effects of lanatoside C on the interaction between Beclin1,VPS34 and VPS15 on the formation of autophagic vesicles in Huh-7 cells were detected by co-immunoprecipitation.Results:The IC50 value of lanatoside C on Huh-7 cells at 24 h was(0.49±0.07)μg/mL.The results of the CCK-8 assay and clone formation assay suggested that lanatoside C had a significant inhibitory effect on the growth of Huh-7 cells in a concentration-dependent manner(both P<0.01).The results of Western blotting assay showed that the LC3-II/LC3-Ⅰ ratio was significantly increased with the increase in the concentration of lanatoside C(P<0.01),while the expression level of Beclin1 protein was up-regulated(P<0.05),and the expression level of p62 protein was significantly down-regulated(P<0.01).The results of laser confocal microscopy showed that lanatoside C could significantly induce autophagic vesicle formation in Huh-7 cells(P<0.01).The results of artificial intelligence image analysis confirmed this effect of lanatoside C(all P<0.01).The results of co-immunoprecipitation confirmed that lanatoside C was able to promote the binding of Beclin1 to VPS34 and VPS15 with each other(both P<0.01).Conclusion:Lanatoside C significantly inhibits the proliferation of Huh-7 cells and and induces Huh-7 cells autophagy,the mechanism of this effect may be related to up-regulating Beclin1 expression and promoting the combination of Beclin1 protein with VPS34 and VPS15 proteins.