Objective:To investigate the expression difference of microRNAs (miRNAs) in urine of chronic hepatitis B (CHB) patients with liver-gallbladder dampness-heat (LGDH) syndrome and liver depression and spleen deficiency (LDSD) syndrome, to explore the characteristic miRNAs of different syndromes and to provide biological markers and objective basis for CHB syndrome differentiation and classification. MethodsUrine samples were collected from 53 subjects with LGDH syndrome, 53 subjects with LDSD syndrome and 24 healthy subjects. The expression levels of urinary miRNA were detected by miRNA microarray and miRNAs with differential expressions were analyzed; The quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to detect the relative expressions of some differentially expressed miRNAs in urine of CHB patients with LGDH syndrome and LDSD syndrome. Prediction, GO and pathway analysis of miRNAs potential target genes were carried out. Receiver-operator characteristic (ROC) curves were generated to determine the specificity and sensitivity of each individual miRNA. The target genes of miRNAs were predicted and Gene Ontology (GO) and pathways were analyzed using bioinformatics methods.  Results:①The microarray of miRNAs expression profiles showed that 21 miRNAs were up-regulated and 1 miRNA was down-regulated in CHB patients with LGDH compared with LDSD (fold change>1.5, P<0.05). ②The results from RT-qPCR revealed that miR-483-3p and miR-4700-3p were in significantly higher expression (P<0.05) and miR-4745-5p was in significantly lower expression (P<0.05) in the urine of CHB patients with LGDH syndrome compared with those with LDSD syndrome; miR-483-3p was in lower expression (P<0.05) and miR-4745-5p was in higher expression (P<0.05) in the urine of CHB subjects with LGDH syndrome LDSD syndrome and miR-4700-3p was in lower expression (P<0.05) in the urine of CHB subjects with LDSD syndrome compared with healthy subjects. ③ROC curve analysis exhibited the AUC (0.8106) and showed that these three miRNAs of miR-483-3p, miR-4700-3p and miR-4745-5p were sensitive and specific enough to distinguish LGDH and LDSD subjects. ④Bioinformatics methods showed that the difference of syndromes expressed potential target genes regulated by miRNAs. GO was mostly related to biological regulation, metabolic process, development of multicellular organisms, stress response and cell proliferation; Pathway was mostly related to post-translational protein modification, TGF-β signaling pathway, TGF-β family member signaling, TGF-β receptor complex signaling and other pathways.  Conclusion:The miRNAs in the urine of CHB patients with LGDH syndrome and LDSD syndrome are differentially expressed. miR-483-3p, miR-4700-3p and miR-4745-5p may be potential markers for syndrome differentiation in CHB and these miRNAs may affect the development of syndromes by regulating their related target genes.