The Chinese excellent traditional culture is the cultural basis of Marxism sinicization. The cultural characteristics and the style of Huxiang culture have made a profound impact on the formation of Mao Zedong Thought. Seeking truth from facts, the mass line and the independence are all the living soul of Mao Zedong Thought, which has a close relation with the idea of practical utility, the traits of sages and heroes, and the idea of depending on oneself in Huxiang culture. A new study of the living soul of Mao Zedong Thought based on the researches of Huxiang culture can expand the research visions of Marxism sinicization, modernization and popularization, and have important values to explore the regional characteristics of the localization of Marxism in China.