【Abstract】Objective To observe serum levels of neuronspecific enolase (NSE) and 25 hydroxy vitamin D(25(OH)D) and explore its clinical significance. Methods Serum levels of NSE and 25(OH)D in 65 cases childhood ALL (including 37 newly diagnosed cases,17 complete remission cases, and 11 relapsed cases) and 35 health children were measured by enzyme linked immunosorbent assay (ELISA). Correlation between serum levels of NSE and 25(OH)D in newly diagnosed cases and its pathological characteristics was analyzed. Serum levels of NSE and 25(OH)D in newly diagnosed cases was analyzed by sequential observation. Results Compared to control group and complete remission group, serum level of NSE was obviously high, while 25(OH)D was remarkably lower, in newly diagnosed cases and relapsed cases (P＜0.01). Serum levels of NSE and 25(OH)D of newly diagnosed cases were irrelevant to age, gender, FAB classification, hepatomegaly, and lymphadenectasis (P＞0.05), and were closely relevant to white blood cell(WBC) of peripheral blood and risk classification (P＜0.01). After complete remission, serum level of NSE was evidently decreased, while 25(OH)D was increased, by sequential observation(P＜0.01). Conclusion Serum levels of NSE and 25-(OH)D has a certain clinical value for occurrence, development, and prognosis evaluation of childhood ALL.