Periventricular leukomalacia (PVL) in premature infants is a major cause of mortality and disability. However, while China is the largest developing country, there is an absence of data concerning PVL. This study was carried out in order to explore the incidence rate and analyze the high-risk factors of PVL in premature infants, and to suggest a working protocol for the prevention of PVL in newborns at risk. The cohort prospective study included 921 premature infants < 37 weeks' gestation from January 2004 to July 2007. The study group was comprised of 271 premature infants with an ultrasound diagnosis of PVL and the control group was comprised of 650 premature infants who were submitted to the same evaluation protocol but did not show any echographic signs of PVL. The results showed that the incidence rate of PVL was 29.4% in China. High-degree PVL accounted for < 12% and > 88% of PVL was low-degree. The incidence rate was much higher when premature infants were accompanied by the following complications: low gestational age, low birth weight, prolonged rupture of membranes, chorionitis, mechanical ventilation, periventricular-ventricular hemorrhage, hypocapnia, and hyperlactacidemia. Severe complications that occur in premature infants also significantly increase the incidence of PVL. Antenatal corticosteroids can significantly decrease the incidence rate of PVL. This investigation provided a theoretical reference for the prevention of premature PVL.