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Objective To determine whether the apoptosis of PC12 cells induced by MPP+ can be protected when the activity of cyclin-dependent kinase 5(CDK5)/p25 is inhibited specifically by TFP5.Methods The 100 μg/L of beta nerve growth factor (β-NGF) was used to induce PCI2 cells differentiating into dopaminergic neurons in vitro.Different concentrations of MPP+ (0,100,200,300,400,600,800 and 1000 μmol/L) were added to the cells;CCK8 assay was used to determine the cell activities and adequate concentration of MPP+.After induction,four groups were designed:PBS and PBS group,MPP+ and PBS group,MPP+ and TFP5 group,and MPP+ and Roscovitine group.Pretreatment of TFP5 and Roscovitne for 12 h was given to the MPP+ and TFP5 group and MPP+ and Roscovitine group,respectively.Hochest33258 staining and flow cytometry were used to detect the cell apoptosis.Western blotting was used to detect the protein expressions ofp35/25,caspase3,cleaved caspase3.Results CCK8 assay showed that the survival rate of PC12 cells was (64.84±1.58)％ when the MPP+ concentration was 300 μmol/L.Flow cytometry indicated significant differences in the apoptosis rate between different groups,which was the highest in MPP+ and PBS group ([25.61±2.74]％),following by MPP+ and TFP5 group ([13.33±1.24]％),MPP+ and Roscovitine group ([9.94±1.70]％),and PBS and PBS group ([8.68±0.21]％);significant difference was noted between MPP+ and TFP5 group and MPP+ and Roscovitine group (P＜0.05).Hochest33258 staining indicated the most obvious nucleus condensation and fragmentation and more apoptotic bodies in MPP+ and PBS group,While few apoptotic bodies were found in MPP+ and TFP5 group and MPP+ and Roscovitine group.Western blotting showed that as compared with that in the PBS and PBS group,the p25 protein level in the MPP+ and PBS group,MPP+ and TFP5 group,and MPP+ and Roscovitine group was significantly increased (P＜0.05).The cleaved caspase-3 protein expression in the MPP+ and PBS group was significantly higher than that in the PBS and PBS group (P＜0.05);the cleaved caspase-3 protein expression in the MPP+ and PBS group was significantly higher than that in the MPP+ and TFP5 group (P＜0.05).Conclusion TFP5 has protective effect against the apoptosis of PC12 cells induced by MPP+ through inhibiting the CDK5/p25 expression and reducing the cleaved caspase-3 protein production.
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Objective To explore the clinical features,laboratory test results,diagnosis,treatment and prognosis of patients with anti-N-methyl-D-aspartic acid receptor (NMDAR) encephalitis.Methods A retrospectives analysis of clinical characteristics,laboratory test,treatment and prognosis of 3 patients with NMDRA encephalitis,admitted to our hospital from February 2012 to November 2014,was performed in our study.Results The most predominant symptoms of 3 patients with anti-NMDAR encephalitis included fever,psychiatric behavioral disturbances,seizures,orofacial dyskinesias and movement disorders and autonomic dysfunction.One patient had immature ovarian teratoma and one had mature mediastinal teratoma.Anti-NMDAR antibody was positive in serum and cerebrospinal fluid of two patients with teratoma,and anti-NMDAR antibody was positive in cerebrospinal fluid and negative in serum of one patient without tumour;all three patients showed abnormal electroencephalography,and 2 had increased signal on MR imaging T2 sequences or fluid-attenuated inversion.Patients were responsive to early immunotherapy and tumour resection.Conclusion The patient with NMDAR encephalitis presented with characteristic clinical manifestations;detection of anti-NMDAR antibody in serum and cerebrospinal fluid may be important for early diagnosis;early immunotherapy and tumour resection are effective for patients with NMDAR encephalitis.