Punctate white matter lesion (PWML) was the common abnormal findings by MRI in neonates, which might predict poor motor and cognitive outcomes. Assisted vaginal delivery (AVD) increased brain injury. But the association between AVD and PWML is not clear. This study showed the mode of delivery is associated with PWML. AVD markedly increased the risk of PWML compared with cesarean delivery, especially vacuum extraction with cesarean section as a reference. The incidence of II and III-IV grade PWML of neonates with AVD was much higher than that with cesarean section.