Background: Chronic hepatitis C (CHC) infection has a close association with diabetes mellitus. However, Conflicting results exist on the analysis of relationship between serum glucose levels and development of fibrosis. The aim of the present study was to evaluate the impact of serum glucose levels on the longitudinal development of liver fibrosis. Methods: In this prospective cohort study, a total of 581 CHC patients were followed from September 2012 to September 2014. Advanced liver fibrosis was defined as a The fibrosis-4 index (FIB-4) index score above 3.25. We assessed the prevalence of liver cirrhosis and identified related factors. We also examined the association between hyperglycaemia and longitudinal development of advanced liver fibrosis. Results: Among 581 patients, 321 (55.2%) showed no advanced fibrosis, 168 (28.9%) showed progression of advanced fibrosis, 81 (13.9%) showed persistent advanced fibrosis and 11 (3.4%) showed improvement at the final visit. Compared to patients with no advanced fibrosis, older age (OR=2.77, 95% CI: 1.85-4.14), heavy alcohol consumption (OR=2.03, 95% CI: 1.22-3.37), and high HCV RNA viral load (OR=3.01, 95% CI: 1.82-4.97) were positively associated with progression of advanced fibrosis. Older age (OR=4.57, 95% CI: 2.58-8.08), hyperglycaemia (OR=3.53, 95% CI: 1.69-7.38), and high HCV RNA viral load (OR=2.44, 95% CI: 1.28-4.65) were independently associated with increased risk of persistently advanced fibrosis. Conclusions: Hyperglycaemia was not substantially associated with progression of advanced fibrosis in shorter-term, whereas hyperglycaemia was positively associated with persistent advanced fibrosis. Strict control of blood glucose levels should be considered for patients with cirrhosis and diabetes.