Objective: This study aims to explore the influence of birth order and age difference on the emotional and behavioral problems among children from two-child family, so as to provide advice on parenting. Methods: A total of 482 students in grades 2 to 5 from the 5 primary schools in Guangzhou were selected, all of whom came from two-child families (excluding twin children). Emotional and behavioral problems were investigated by using the Conner’s Parental Symptom Questionnaire (PSQ). Results: Among children from two-child families, higher prevalence of behavioral problems, learning problems, impulsive hyperactivity and hyperactivity index in older compared with younger children (β=-0.09，-0.16，-0.09，-0.10,P<0.05). The scores of behavioral problems and hyperactivity index were significantly higher among those of children who were 6-9 years older than young siblings, compared to those who were 0-3 years older than younger siblings(β=0.20,0.21,P<0.05). No similar pattern was found among the younger siblings(P>0.05). Conclusion Among school-age children from two-child families, emotional and behavioral problems of the elder ones were more serious than the younger ones’. Age difference between siblings shows impact on the emotional and behavioral development among elder children, however no significant impact is noticed among the younger ones.