overexpression of CIRP and its mechanisms research Cold inducible RNA-binding protein (CIRP) is a glycine-rich RNA-binding protein with an RNA recognition motif and a carboxyl-terminal domain. CIRP initially localized in the nucleus, under some stresses, especially cold stress, it can translocate to the cytoplasm and bind the 3’ regions of the target mRNA with its RNA-recognition motif, and then exerts an effect on regulating gene expression. CIRP expression in the brain is increased significantly under cold stress, and the upregulation of CIRP plays an important role on triggering the animal hibernation. These findings inspired the idea that highly expressed CIRP may induce neuroprotection. Our previous studies demonstrated that CIRP can inhibit neuron apoptosis, overexpressed CIRP under hypoxia can restore the proliferation of neuronal stem cells, suggesting a neuroprotective role of CIRP. We hypothesized that highly expressed intracellular CIRP with minimal extracellular distribution could achieve a most positive effect on neuroprotection. Then the pathway underlying intracellular CIRP expression may provide new clue for a practical neuroprotection protocol. CIRP overexpression was established on a glial cell line, SHG 44, through transfection of lentivirus with CIRP. . The effects of CIRP overexpression on the cell viability were evaluated by MTT assay after treatment with oxidation, hypoxia, neurotoxicity, and glucose deprivation. To explore the relationship between highly expressed CIRP and some neuroprotective factors, the mRNA expressions of these factors were analyzed by Real-time polymerase chain reaction. Then Double immunohistochemistry and western blot were used to determine the increased expressions.