AIM: To investigate the prevalence and clinical effects of the comprehensive treatment of demodex in eyelash follicles in patients with Meibomian gland dysfunction(MGD). METHODS: This study was a prospective analysis.The MGD patients(experimental group, 264 eyes in 132 patients)and normal subjects(control group, 192 eyes in 96 patients)were collected from the He Eye Hospital from January 2016 to July 2017. Comprehensive treatment of patients in the experimental group. Eyelash sampling, demodex counting, tear film break up time(BUT), cornea fluorescein staining(FL)and Schirmer Ⅰ test(SⅠt)were measured for both groups. The symptom score of MGD patients was recorded.RESULTS: For the experimental and control groups, the number of demodex before the treatment was 7(6, 9)and 2(1, 3), respectively(Z=5.264, P<0.01). The percentage of demodex infestation was 100% and 28.1%,respectively(χ2=35.957, P<0.01). The BUT was 4.06±1.38 and 12.00±2.82s. The FL was 3.06±1.57 and 0.46±0.63. The SⅠt was 6.93±2.08mm and 11.13±2.38mm/5min, respectively(t=-9.825, t=5.978, t=-4.776; all P<0.01). The symptom score of the MGD group before and after treatment was 6.57±2.93 and 3.27±1.89, respectively(t=5.443, P<0.01). After the treatment, the percentage of demodex infestation of the experimental group were 57.6%, the number of demodex were 3(0-5), had significant difference with the data before the treatment(Z=3.937, P<0.01). The BUT and FL were 6.53±3.27s and 1.67±0.54, and all had significant difference with the data before the treatment(t=5.152, 4.328; all P<0.01). The SⅠt of the experimental group after the treatment was 8.37±5.34mm/5min, with no significant difference with the data before the treatment(t=-0.285, P=0.748).CONCLUSION: Demodex infection can cause Meibomian gland dysfunction and produce corresponding clinical symptoms. The comprehensive treatment can greatly diminishes the number of demodex mites and effectively relieve the symptoms of Meibomian gland dysfunction which is associated with demodex infestation.