Objective To understand job satisfaction and influence factors among staffs in 12320 health hotline. Methods Job satisfaction questionnaire was designed, all staffs in 12320 health hotline selected from representative province in our country finished the questionnaire alone. Descriptive statistics was done, and logistic regression method was used to analyze satisfaction influence factors. Results The average scores of comprehensive assessment job satisfaction about staffs was 3.96±0.70, the highest dimension was management system, but the scores of interpersonal relationship and job return was low. The influence between working time and comprehensive assessment job satisfaction had statistical significance, similarly employ way and working environment satisfaction, job position and job itself satisfaction, age and average monthly income to job return satisfaction had statistical significance. Job satisfaction had negative correlation with turnover intention. Conclusion The level of job satisfaction about staffs in 12320 health hotline was neither high nor low. Some effort should be taken to improve job satisfaction level, such as raising the salary and stable staffs team.