Abstract：Objective To explore the application effect of ultrasound assisted combined with vascular puncture map in optimizing the puncture protocols for newly created autologous arteriovenous fistula （AVF）.  Methods A total of 80 patients undergoing maintenance hemodialysis at the hemodialysis center of the Nanning First People's Hospital from January 2022 to December 2022 were selected and divided into a control group and an observation group using a random number table method, with 40 patients in each group. The control group underwent conventional blind cannulation, while the observation group underwent ultrasound assisted cannulation guided by a pre-drawn vascular puncture map. The success rate of first-attempt cannulation, hemodialysis blood flow velocity, and the incidence of AVF-related complications between two groups were compared. Results In the observation group, both the cannulation success rate during the AVF initiation phase and at 6 months post-creation were significantly higher than those in the control group, there was a statistically significant difference between the two groups （χ2＝8.889, 7.671, both P＜0.05）. At 6 months, blood flow velocity during hemodialysis was significantly greater in the observation group than in the control group, there was a statistically significant difference between the two groups （t＝7.383, P＜0.05）. The incidence rates of vatious complications retated to AVF were significantly lower in the observation group （χ2＝8.889, 4.114, 8.650, 6.486; all P＜0.05）. Conclusion The application of ultrasound assisted combined with vascular puncture imaging substantially improves first-attempt cannulation success rates for newly created autologous AVF and reduces the incidence of AVF-related complications.