Objective:To evaluate the impact of the sequence of pulmonary vein ablation during cryoablation on vagal reflex and heart rate variability(HRV).Methods:A retrospective analysis was conducted on 114 paroxysmal atrial fibrillation patients who underwent cryoballoon ablation at our institution from January 1,2020 to June 30,2023.Patients were divided into two groups based on the sequence of cryoablation:the firstly right superior pul-monary vein ablation group(RSPV,n=57)and the firstly left superior pulmonary vein ablation group(LSPV group,n=57).Vagal reflex events and heart rate variability indicators between the two groups were compared.Results:During cryoablation,the RSPV group had a significantly lower incidence of vagal reflex compared to the LSPV group(28.1%vs 1.8%,P<0.000 1).Only one case of vagal reflex,manifested as sinus bradycardia,oc-curred in the RSPV group,while 16 occurred in the LSPV group manifested as sinus bradycardia,atrioventricular block,cardiac arrest(all patients required pacing),and hypotension.Heart rate variability indicators SDNN,SDANN,rMSSD,and LF/HF decreased significantly after ablation compared to those before ablation in the LSPV and RSPV groups(all P<0.05),but there was no statistically significant difference in the above indicators between the two groups.Conclusion:Starting cryoablation from the right pulmonary vein,followed by the left pul-monary vein,significantly reduces the occurrence of vagal reflex during atrial fibrillation cryoablation.While HRV decreases significantly after ablation,the change in HRV is not related to the sequence of ablation.