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Objective:To explore the effect of intragastric administration of diphenoxylate on inducing epilepsy in mice, as well as the changes in neurobehavioral and hippocampal neurons in mice.Methods:Forty C57 male mice aged 2-3 weeks were selected and divided into control group and diphenoxylate group using random number table method, with 20 mice in each group. The mice in the diphenoxylate group were given diphenoxylate (200 mg/kg) by gavage once a day for 14 consecutive days, while the control group mice were given an equal volume of 0.9% sodium chloride solution every day. After each gavage, the seizure status of mice within 2 hours was observed and the mice were graded based on the Racine score. Open field test, elevated cross test, and Morris water maze test were used to observe the neurobehavioral activities of mouse.A digital electroencephalogram machine was used to monitor the epileptic seizures of mice induced by diphenoxylate.Nissl staining was used to observe the damage of hippocampal neurons in mice.SPSS 25.0 software was used for statistical analysis of the data, and independent sample t-test was used for pairwise comparison. Results:The Racine grading results showed that the mice in diphenoxylate group exhibited grade 2 and 3 seizures at 1 hour after gavage. The EEG monitoring results showed that compared with before gavage, the frequency and amplitude of brain waves of mice in diphenoxylate group increased.In the open field test, the residence time in the central region of mice in diphenoxylate group was significantly lower than that of the control group ((12.21±3.37)s, (17.05±4.34)s, t=3.29, P<0.01). In the elevated cross test, the residence time in the open arm of mice in diphenoxylate group was significantly lower than that of the control group ((17.36±5.41)s, (26.70±9.06)s, t=3.31, P<0.01). In the Morris water maze test, the residence time in the platform quadrant of diphenoxylate group was significantly lower than that of the control group((22.08±6.76)s, (27.64±4.60)s, t=2.54, P<0.05). The residence time and the number of stays in the platform area of diphenoxylate group were both significantly lower than those of the control group(both P<0.05). Nissl staining showed that the number of hippocampal neurons in the CA3 region of mice treated with diphenoxylate was significantly lower than that in the control group((135.67±4.59), (140.67±2.73), P<0.05). Conclusion:Excessive diphenoxylate can induce seizures in mice, and the mice exhibit increased anxiety-like behavior and decreased spatial learning and memory abilities.
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Objective:To investigate the expression of N-myc downstream regulated gene 2(NDRG2) in hippocampus of epileptic mice and its effect on glutamate and glucose uptake in astrocytes of mice.Methods:The epileptic mouse model was induced by lithium chloride and pilocarpine nitrate. The mice were sacrificed at 1 d, 7 d, 15 d and 6 weeks after model establishment and the brain tissues of hippocampus were taken. Western blot was used to detect the expression of NDRG2 protein in hippocampus.The primary astrocytes of wild-type, NDRG2 + /+ and NDRG2 -/- mice were cultured and the NDRG2 phenotype of astrocytes was identified after primary culture. Glutamate content in the supernatant of astrocyte culture was determined by glutamate assay kit and ultraviolet spectrophotometer. Flow cytometry was used to detect the positive rate of 2-NBDG fluorescently labeled astrocytes. Results:(1) Compared with the control group (0.25±0.07), the expression of NDRG2 in the hippocampus of mice increased significantly in the acute phase of epilepsy (1 d(0.45±0.06, t=-3.84, P<0.05), 7 d(0.54±0.09, t=-4.30, P<0.05), 15 d(1.04±0.06, t=-15.08, P<0.01)), and remained significantly high in the chronic phase of epilepsy( 6 weeks (1.30±0.16, t=-10.40, P<0.01)). (2) The content of residual glutamate in the supernatant fluid of primary cell culture medium was detected.It was found that the uptake of glutamate by astrocytes in the NDRG2 -/- group was significantly lower than that in the NDRG2 + /+ group ((689.03±101.78) μmol/L, (113.67±37.35) μmol/L; t=9.19, P<0.01). (3) Western blot results showed that the expression of EAAT1 protein in NDRG2 -/- primary astrocyte was significantly lower than that of NDRG2 + /+ primary astrocyte(0.34±0.03, 1.16±0.21), and the difference was statistically significant ( t=-6.59, P<0.01). (4) Flow cytometry results showed that the positive rate of astrocyte in NDRG2 -/- group cells was significantly lower than that in NDRG2 + /+ group cells ((17.60±5.72)%, (72.22±8.35)%), and the difference was statistically significant ( t=-13.22, P<0.01). Conclusion:NDGR2 is closely related to the occurrence and development of epileptic diseases. The expression of NDRG2 is beneficial to exert its physiological function of EAAT1 and promotes the uptake of glutamate and glucose by astrocyte. It may be a potential cell protective factor to promote nerve protection and repairment.
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BACKGROUND: There are various kinds of design methods about preoperative osteotomy of ankylosing spondylitis with kyphosis, but each has their own errors and limitations. A convenient, precise and available method needs to bedeveloped.OBJECTIVE: To compare the clinical efficacy of two different preoperative osteotomy designs using paper-cut andphotoshop software for ankylosing spondylitis with kyphosis.METHODS: Thirty-nine patients suffering ankylosing spondylitis with kyphosis undergoing osteotomy in the Departmentof Spinal Surgery, Affiliated Hospital of Yan'an University between June 2009 and January 2015 were enrolled, andrandomly allotted to paper-cut (n=19) and photoshop (n=20) groups, followed by the preoperative osteotomy design,respectively. All patients were followed for 12-40 months to compare the postoperative osteotomy angle error andcorrection efficacy at the last follow-up between groups.RESULTS AND CONCLUSION: (1) The postoperative osteotomy angle error in the photoshop group was significantly smaller than that in the paper-cut group (P 0.05). (4) To conclude, compared with the osteotomy design usingtraditional paper-cut splice, the photoshop software can achieve a smaller osteotomy angle error and better postoperative balance of spinal sagittal plane, thus providing precise osteotomy for surgeons to obtain proper correction.