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Objective To evaluate the effect of glandular involvement and persistent human papillomavirus（HPV） infection upon the recurrence of high-grade squamous intraepithelial lesion（HSIL） of the cervix， aiming to provide evidence for postoperative clinical management. Methods Clinical data of 596 HSIL patients who underwent cold-knife conization were collected， including 240 cases of glandular involvement（glandular involvement group） and 356 cases of non-glandular involvement（non-glandular involvement group）. The recurrence of HSIL at postoperative 2 years was considered as the end point. The differences between recurrent and non-recurrent patients were compared. According to glandular involvement and HPV-16 infection， patients were divided into the glandular involvement+HPV-16 infection， glandular involvement alone， HPV-16 infection alone and non-glandular involvement+non-HPV-16 infection groups， respectively. The differences in the recurrence rate of HSIL were compared among four groups. Results Among 596 HSIL patients， 75 cases recurred（12.6%）. HPV-16 infection rate in the glandular involvement group was 67.9%， significantly higher than 23.0% in the non-glandular involvement group， and the difference was statistically significant（P < 0.05）. In univariate analysis， the proportion of≥45 years old， glandular involvement and persistent HPV-16 infection were not significantly correlated with the recurrence of HSIL（all P > 0.05）. The recurrence rates of HSIL were 33.8%， 10.9%， 8.5% and 9.7% in the glandular involvement+HPV-16 infection， glandular involvement alone， HPV-16 infection alone and non-glandular involvement+non-HPV-16 infection groups， respectively. The recurrence rate of HSIL in patients with glandular involvement+HPV-16 infection was significantly higher than that in other three groups， and the difference was statistically significant（all P < 0.017）. Conclusions HSIL patients with glandular involvement and persistent HPV-16 infection are prone to relapse after cervical cold-knife conization. For these patients， active surgical regimen should be adopted and postoperative follow-up should be strengthened.