To investigate the efficacy and prognostic factors of cervical cold knife conization (CKC) combined with recombinant human interferon α-2b gel for treating patients with grade II of cervical intraepithelial neoplasia (CIN). Methods: A total of 132 patients with CIN grade II were selected as the research subjects from January 2019 to January 2021. These patients all were treated by CKC combined with recombinant human interferon α-2b gel. The curative effect, intraoperative blood loss, hospital stay, disappearance time of the vaginal excessive secretion, bloody leucorrhea disappearance time, and CIN recurrence rate of these patients were observed. The changes of IL-10, CRP, TNF-α, and other inflammatory factor levels of the patients were compared between before and after treatment. These patients were divided into group A (102 patients with CIN recurrence) and group B (30 patients without CIN recurrence) according to the CIN recurrence situation during the follow-up period. The risk factors affecting the prognosis of the patients were analyzed. Results: The total effective rate of 132 patients was 96.2%, their intraoperative blood loss was 10.5±1.3ml, their time of hospital stay was 4.2±1.5d, their disappearance time of vaginal excessive secretion was 5.6±1.3d, and their disappearance time of hemorrhagic leucorrhea was 5.3±1.5d. The inflammatory factors of the patients had been improved after operation (P＜0.05). The CIN recurrence rate of these patients was 22.7% after 12 months of follow-up. There were no significant differences in the body mass index (BMI) value, the history of alcohol, the intraoperative blood loss, the number of pregnancies, and the number of parities of the patients between group A and B (P＞0.05), but there were significant differences in the age, the situations of menopause and surgical margin, the smoking history, the human papillomavirus (HPV) infection situation, the degree of cervical erosion, the history of abortion, and the initial age of sexual life of the patients between group A and B (P＜0.05). Binary logistic regression analysis showed that age ≥55 years old, the menopause status, the positive lesion of surgical margin, the smoking history, the high-risk HPV infection, the severe cervical erosion, the abortion history, the initial age of sexual life <16 years old of the patients were the independent risk factors affecting their prognosis of grade Ⅱ CIN (P＜0.05). Conclusion: CKC combined with recombinant human interferon α-2b gel for treating patients with CIN grade II has ideal effect. The age, the menopause status, the surgical margin situation, the smoking history, the HPV infection, the severe cervical erosion, the abortion history, the initial age of sexual life of the patients are the risk factors affecting their prognosis.