Objective To explore the changes of serum thyroid stimulating hormone (TSH) and thyroglobulin (Tg) levels and their clinical value in the diagnosis of malignant and benign thyroid nodules. Methods The serum levels of TSH and Tg in 96 patients with thyroid nodular disease were measured, the relationship between the serum TSH andTg levels of patients with benign and malignant thyroid nodules were analyzed. Results The serum TSH levels in patients with thyroid carcinoma and with benign thyroid nodules were 2.55 ± 1.46 mIU/L and 1.15 ± 1.10 mIU/L respectively and had significantly different (P<0.05). The ratio of thyroid nodule canceration was increase as the increase of serum levels of TSH in patients with thyroid nodular disease. The serum Tg levels in thyroid cancer patients were significantly higher than that in patients with benign thyroid nodules (P<0.05). There was no significantly difference in the malignant thyroid nodule numbers between different serum Tg levels (P>0.05). Conclusion The detection of serum TSH and Tg has certain clinical significance in the diagnosis of malignant and benign thyroid nodule, and serum TSH level might be used as a factor in prediction of thyroid nodule canceration.