Abstract：Objective Research presented by this paper reveals the trend in drug resistance of Gram-positive coccus and thus provides guidance to effective use of antibiotics in the medical treatment. This research was based on clinical data of isolation and drug resistance of Gram-positive coccus collected from clinical patients and in-patient in a regional hospital during the period of 2011—2016. Methods In this research, we applied isolation and culturing to samples of the Gram-positive coccus bacteria collected from patients treated by the hospital between January 2011 and December 2016. We used both fully automated identification system and and API manua identification system for bacterial identification. The drug sensitivity test and results were determined by referencing to clinical and laboratory standards institute (CLSI), m100-s26. The Whonet 5.6 software and SPSS 13.0 software were used for the statistical analysis. Results From 2011 to 2016, out of all 50,412 strains of isolated bacteria in the hospital, 15123 of them were Gram-positive coccus, approximately 29.99% of the total count. Staphylococcus aureus, Streptococcus pneumoniae, Staphylococcus epidermis, Enterococcus faecalis, Enterococcus faecium, β-hemolytic Streptococcus, and Streptococcus viridans group ranked among the top seven, and were 6.48%~7.78%, 5.05%~6.47%, 1.25%~6.47%, 0.91%~1.26%, 0.62%~1.26%, 0.55%~1.05%, and 0.60%~1.05% of the total bacteria, respectively, and showed no statistical difference (P>0.05). The trend chart showed that there was a gradual upward trend in the separation proportion of β-hemolytic Streptococcus and Streptococcus viridans. From 2014 to 2016, the percentage of Enterococcus faecalis had been decreasing while that of Enterococcus faecium increasing. In 2016, the percentage of Enterococcus faecium became greater than that of Enterococcus faecalis. The MRSE ratio was between 52.30%~77.60%, and the proportion of MRSA was between 32.20%~46.80%, and the proportion was increasing year by year. The proportion of Streptococcus pneumoniae resistant to penicillin was between 1.70% and 3.10%. This research did not find Staphylococcus aureus or Staphylococcus epidermidis that was resistant to either vancomycin or linezolid. The resistance of Staphylococcus aureus to tetracycline, ciprofloxacin, and levofloxacin had a declining trend to a certain degree. Drug resistance of Enterococcus has been relatively stable. Streptococcus pneumoniae has a higher resistance rate to erythromycin, clindamycin, trimethoprim/sulfamethoxazole and tetracycline. The resistance rate of the Streptococcus viridans to ampicillin and cefotaxime increased year by year, and β-hemolytic Streptococcus has kept a high sensitivity to common antibiotics, but resistant strains to levofloxacin were also found in recent years. Conclusion The separation ratio of Gram-positive bacteria was relatively stable, and the proportions of β-hemolytic Streptococcus, Streptococcus viridans and Enterococcus faecium were increasing year by year. MRSA rate observed during this research was above national average level, while MRSE rate was lower than national average, both with rising trends. PRSP and VRE remained at low levels throughout the country. Drug resistance of Staphylococcus aureus was unstable with a great variance, drug resistance of Staphylococcus epidermidis was relatively stable. Two kinds of staphylococci had a lower drug resistance rate for fluoroquinolones, and there was a decreasing trend year by year. The resistance rate of Streptococcus pneumoniae to cefotaxime increased year by year. The multidrug resistant Streptococcus viridans were increasing year by year. Extensive study of the drug resistance monitoring data for the hospital and its covered geographical area is very useful to identify characteristics of drug resistance of bacteria in the area, and thus provides effective guidance to the clinical use of antibiotics.