Objective: To investigate and summarize the measures for identification and protection of parathyroid glands in total thyroidectomy (TT). Methods: The clinical data of 3 425 patients who underwent TT from March 2003 to March 2013 were analyzed retrospectively. Results: Of the entire group of patients, 489 cases were male and 2 936 cases were female, and their ages ranged from 9 to 72 years; operation was performed in 2 261 cases of nodular goiter, 684 cases of thyroid cancer, 92 cases of primary hyperthyroidism, 57 cases of hyperthyroidism resulting from a nodular goiter, 41 cases of thyroid adenoma, 31 cases of granulomatous thyroiditis, and 259 cases of Hashimoto’s disease; 364 cases underwent unilateral TT and bilateral TT was done in 3 061 cases; 186 cases underwent neck dissection and 834 cases received parathyroid autotransplantation. The length of postoperative hospital stay was 5 to 8 d and postoperative follow-up period ranged from 2 months to 10 years; there were no cases of permanent hypoparathyroidism and the efficacy of treatment was satisfactory. Conclusion: The parathyroid glands have special anatomical features, with complicated and diverse shapes. During TT, surgeons should operate meticulously, dissect along the tissue planes, and keep a clean operative field so as to accurately identify and protect the parathyroid glands and avoid damaging them, which may effectively reduce the incidence of permanent hypoparathyroidism.