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Diffuse Axonal Injury (DAI) is the most common type of traumatic brain injury, and it is associated with the linear and rotational accelerations resulting from head impacts, which often occurs in traffic related and sports accidents. To investigate the degree of influence of linear and rotational acceleration on DAI, a two-factor, two-level rat head impact experimental protocol involving linear and rotational acceleration was established using the L4(23) orthogonal table in this paper. Following the protocol, rats head was injured and diffusion tensor imaging (DTI) was performed at 24h post-injury to obtain the whole brain DAI injury, and the fractional anisotropy (FA) value of the corpus callosum was selected as the evaluation indicator. Using analysis of variance, the sum of squared deviations for the evaluation indicators was calculated to determine the degree of influence of linear acceleration and rotational acceleration on DAI. The results show that, 1.