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Secondary spinal cord injury is the inflammatory damage to surrounding tissues caused by activated microglial-mediated neuroinflammatory responses. The nuclear factor-κB (p65/p5 pathway and PU.1 are closely correlated with inflammatory responses; thus, we examined the relationship and function between PU.1 and p50 in secondary spinal cord injury. In this study, we established an adult rat acute spinal cord injury model to simulate the pathological process of spinal cord injury. We found that the expression of PU.1 was significantly inc