Patient-centered nursing holistic care is of utmost important to the nursing profession, and humanistic care cultivation has become a global nursing education concern. This study aimed to examine the relationship among emotional intelligence, empathy and humanistic care ability in nursing students, and to determine whether positive emotional intelligence could mediate the relationship between empathy and humanistic care ability. This study used a cross-sectional, descriptive design. A total of 323 nursing students was enrolled from one medical university in Heilongjiang Province, China. The emotional intelligence questionnaires, empathy scale and humanistic care ability scale were used to quantify participants’ responses. There was no significant difference in gender, residence, single-child family and leader experience of nursing students’ humanistic care ability. However, the significant differences were found in grade (t = 4.55, P < .01) and major interests (t = 7.06, P < .01). Obviously, there was positive correlation between positive emotional intelligence and empathy (R = 0.37, P < .01), and positive correlation between humanistic care ability and emotional intelligence (R = 0.62, P < .01), and empathy (R = 0.57, P < .01). Furthermore, emotional intelligence (β = 0.21, P < .01) had a significant mediating effect on the relationship between humanistic care ability and empathy. Nurse educators should improve the empathy of nursing students by developing and implementing emotional intelligence programs, in order to improve their humanistic care ability.