Objective: To investigate the relationship between age, gender and the change of the pupil diameter and the rate of response in patients with general anesthesia. Methods: Selected 90 patients (180 eyes) with general anesthesia from October to July 2015 in General Hospital of Lanzhou Army. Divided into low age group (3-40 years old), the middle age group (41-60 years old), and high age group (over 61 years old). The low age group of 30 people (60 eyes), including 16 males (32 eyes), 14 females (28 eyes), average age (22.60±10.36) years old; the middle age group of 30 people (60 eyes), 13 males (26 eyes), 17 females (34 eyes), average age (48.73±5.82) years old; high age group of 30 people (60 eyes), 20 males (40 eyes), 10 females (20 eyes), average age (66.63±4.88) years old. Results: Before and after anesthesia there were significantly differences between the left and right eye in each group (P<0.05). Before anesthesia there was no significant difference between male and female, after anesthesia there was no significant difference between male and female in each group. Compared either two groups of low, middle, and high age, it is found that, with the increase of age, the pupil diameter and the reaction rate of patients has statistical significance (P<0.05). Conclusion: Patients with general anesthesia for pupil diameter changes with age, gender, closely observe patient’s pupil and the reaction rate change on the anesthesia effect, the change of intraoperative patients has guiding significance.