To investigate the value of free beta subunit of human chorionic gonadotropin (F-β-hCG) detection combined with ultrasound intracranial translucency (IT) thickness for screening neural tube malformation (NTDs) during the second trimester of pregnancy. Methods: 54 pregnant women with confirmed NTDS fetuses were selected in observation group from July 2015 to September 2019, and 68 pregnant women with normal fetuses were selected in control group during the same period. The F-β-HCG level of the pregnant women, and the values of fetal IT and nuchal translucency (NT), and crown-rump length (CRL) during the second trimester of pregnancy were measured. The correlation between the value of fetal IT thickness by ultrasound and F-β-HCG level was analyzed. The values of fetal IT thickness by ultrasound and F-β-HCG level for screening fetal NTDS were also analyzed. Results: The ultrasonic IT thickness (1.29±0.37mm) of the fetus in the observation group was significant lower than that (1.72±0.38mm) of the fetus in the control group, and the F-β-HCG level (213.61±62.64μg/L) of the women in the observation group was significant higher than that (124.30±39.71μg/L) of the women in the control group (all P＜0.05). The area under the ROC curve, the cut-off value, the sensitivity, and the specificity of ultrasonic IT thickness for diagnosing NTDs were 0.842, 1.580, 88.9%, and 63.2%, respectively. The area under the ROC curve, the cutoff value, the sensitivity, and the specificity of F-β-HCG level for diagnosing NTDs were 0.868, 143.252μg/L, 87.0%, and 73.5%, respectively. The area under the ROC curve, the sensitivity, and the specificity of ultrasonic IT thickness combined with F-β-HCG level for diagnosing NTDs by were 0.930, 94.4%, 82.4%, respectively. The F-β-HCG level of the women was negatively correlated with the fetal values of IT and CRL, but was positively correlated with the fetal NT value (P＜0.05). Conclusion: Ultrasounic IT value of the fetus and the F-β-hCG level of the women during the second trimester of pregnancy have certain value for screening NTDs, and the ultrasounic IT of the fetus combined with the F-β-hCG level of the women has higher value for screening NTDs.