Objectives: Heroin dependence (HD) is a chronic brain disease with genetic contribution. NeuroD1/D6 are neurogenic differentiation factors that play a role in development and differentiation of the brain. In this study, we intended to investigate the association of NeuroD1/D6 gene polymorphism and the occurrence of HD, and the role of this genetic variability in specific personality traits of HD patients. Materials and Methods: A total of 694 individuals were recruited in this study (313 HD patients and 381 normal controls). Patients with HD were further classified into four clinical subgroups to reduce heterogeneity. Genetic variants of selected single‑nucleotide polymorphisms in NeuroD1 (rs1801262, rs16867467, and rs2583016) and NeuroD6 (rs2233404) were genotyped. We used the Chinese version of tridimensional personality questionnaire to assess specific personality traits in subjects. Results: For allele and genotype frequency analysis, we found a weak association between NeuroD1 rs16867467 and patients with HD (P < 0.05). Nonetheless, the association findings could not remain significant after Bonferroni’s correction. For specific personality traits, NS score was negatively correlated with age in both study groups (P < 0.05); however, the NeuroD SNPs were not associated with the novelty‑seeking/harm avoidance scores in patients with HD. Conclusions: Our study suggests that NeuroD1/D6 gene polymorphism may neither be associated with the occurrence of HD nor did these polymorphisms influence the personality traits in patients with HD.