The incidence of Papillary Thyroid Carcinoma (PTC) has increased gradually in recent years, especially for Papillary Thyroid Microcarcinoma (PTMC). The purpose of this study was to evaluate the expression of Ki67, CK19, Galectin-3 and HBME-1 protein in PTMC and evaluate the clinical significance of differential diagnosis. The specimens of 80 PTMC cases and 40 benign papillary hyperplasia thyroid cases were collected in our study. Ki67, CK19, Galectin-3 and HBME-1 protein expression were evaluated in these specimens by using immunohistochemistry.The positive expression rates of Ki67, CK19, Galectin-3 and HBME-1 in PTMC were 43%, 99%, 95% and 71%. Respectively, the rates were 15%, 38%, 25% and 15% in benign thyroid (P<0.01). Sensitivity of Ki67, CK19, Galectin-3 and HBME-1 in differential diagnosis were 43%, 99%, 95% and 71%; Specificity were 85%, 63%, 75% and 85%;Diagnostic accuracy were 57%, 87%, 88% and 76%; The area under the ROC curve were 0.638, 0.806, 0.850 and 0.781. Our study suggested that detection of Ki67, CK19, Galectin-3 and HBME-1 protein may be helpful for differential diagnosis of PTMC, and Galectin-3 was the best one in these four markers for differential diagnosis.