Objective To investigate the distribution characteristics of 677C/T polymorphism of methylene tetrahydrofolate reductase (MTHFR) gene in western Guangxi population and to analyze the distribution differences of 677C/T polymorphisms in Guangxi and other regions, and further to explore whether the 677C/T polymorphism is associated with hyperhomocysteinemia (HHcy) and serum homocysteine (Hcy) level. Methods A total of 2 339 people who underwent physical examination and tested for MTHFR gene at the Health Department of Baise People′s Hospital from January 2020 to January 2022 were selected as study subjects. The polymorphism of 677C/T was detected by the PCR-fluorescence probe method and the serum Hcy was detected by the circulating enzyme method. Differences in the distribution of 677C/T polymorphism were compared according to gender and regional characteristics. In addition, the study subjects were divided into the HHcy group and the control group according to the serum Hcy>15 μmol/L defined as HHcy, and then the correlations between MTHFR 677C/T gene polymorphism and HHcy and serum Hcy level were analyzed. Results Among the 2 339 physical examination population in western Guangxi, the frequencies of CC,CT,and TT genotype of MTHFR 677C/T gene were 59. 9%,34. 2%,and 5. 9%,respectively, and the C and T allele frequencies were 77. 0% and 23. 0%,respectively. There were no statistically significant differences in genotype frequency and allele frequency between male and female (P>0. 05) . The distribution of 677C/T gene polymorphisms in western Guangxi was not significantly different compared with the population in Maoming area (P>0. 05),but there were statistical differences compared with the domestic populations in Huizhou, Quanzhou, Wuhan, Gansu, Beijing and Shanxi and foreign populations in Europe, Japan, Africa, Mexico and Indian (P<0. 05) . The results of the genetic susceptibility analysis showed that individuals carrying CT genotype and TT genotype had 1. 60 times (OR=1. 60,95%CI 1. 24-2. 08,P<0. 001) and 5. 21 times (OR=5. 21,95%CI 3. 24-8. 40,P<0. 001) higher risk of developing HHcy than individuals carrying CC genotype, respectively. Individuals carrying T allele had 1. 82 times (OR=1. 82,95%CI 1. 51-2. 19,P<0. 001) higher risk of developing HHcy than individuals carrying C allele. In the HHcy group, the serum Hcy level in individuals carrying TT genotype were significantly higher than those carrying CC genotype and CT genotype with statistically significant differences (P<0. 001) . However, there were no statistical differences in the serum Hcy level among genotypes in the control group (P>0. 05) . Conclusion The distribution of MTHFR 677C/T polymorphism in western Guangxi is not affected by gender, but it has geographically specific. The T allele may be an independent risk genetic factor for developing HHcy in western Guangxi population and the homozygous mutant TT genotype is closely associated with elevated serum Hcy level. [ABSTRACT FROM AUTHOR]