Objective:To investigate the expression of adenosine 5'-monophosphate-activated protein kinase (AMPK) phosphorylation in corneal epithelial cells and the effects of fungus on AMPK phosphorylation and interleukin-6 (IL-6) production in corneal epithelial cells.Methods:The human immortalized corneal epithelial cell line was selected.The safe concentration range of AMPK agonist 5-aminoimidazole-4-carboxamide 1-β-D-ribofuranoside (AICAR) (100, 300, 500, 1 000 μmol/L) and inhibitor Compound C (10.0, 12.5, 15.0, 17.5, 20.0 μmol/L) on corneal epithelial cells was screened by multi-function real-time unlabeled cell analyzer.Corneal epithelial cells without any treatment were used as the normal control group, and those co-cultured with spores were used as the spore control group.Corneal epithelial cells co-cultured with spores were treated with AICAR and Compound C for 4 hours in the AICAR group and Compound C group, respectively.The expression of phosphorylated AMPK (p-AMPK) and AMPK in corneal epithelial cells was detected by Western blot, and the concentration of IL-6 in the culture supernatant was determined by enzyme-linked immunosorbent assay (ELISA).Results:After treatment with different concentrations of AICAR for different periods, there was no statistical significance in the cell index of corneal epithelial cells (all at P>0.05). The cell index of corneal epithelial cells was increased with 10.0 μmol/L and 12.5 μmol/L Compound C treatment compared with that of the normal control group.The expression levels of p-AMPK were 0.67±0.15, 2.57±0.12, 3.67±0.58 and 1.50±0.50, respectively, in the normal control group, spore control group, AICAR group and Compound C group, showing a statistically significant difference among them ( F=32.820, P<0.001). The expression level of p-AMPK was significantly higher in the spore control group compared with the normal control group ( P<0.001). The expression level of p-AMPK in the AICAR group was higher than that in the spore control group, and the expression level of p-AMPK in the Compound C group was lower than that in the spore control group, and the differences were statistically significant (both at P=0.010). There was no significant difference in the relative expression level of AMPK among the four groups ( F=0.120, P=0.950). The expression levels of IL-6 concentration in the normal control group, spore control group, AICAR group and Compound C group were (107.81±17.15), (156.32±9.94), (167.96±14.16) and (127.42±19.75)pg/ml, respectively, showing a statistically significant difference among them ( F=15.210, P<0.001). The IL-6 concentration of the spore control group was higher than that of the normal control group, and the difference was statistically significant ( P<0.001). The IL-6 concentration of the AICAR group was higher than that of the spore control group, but the difference was not statistically significant ( P=0.260). The IL-6 concentration of the Compound C group was lower than that of the spore control group, and the difference was statistically significant ( P=0.010). Conclusions:In corneal epithelial cells, AMPK phosphorylation is found, which is enhanced after fungal spores stimulation, and the secretion of IL-6 increases.

Objective To investigate the effect of scalp site medication injection on endogenous neural stem cells proliferation,differen-tiation and the expression of the nerve regeneration inhibitory factor Nogo-A mRNA in newborn rats with hypoxic-ischemic encephalopathy(HIE).Methods Seven-day-old SD rats were randomly divided into 4 groups: sham operation group(n = 20),HIE model group(n=20) and sites injected with normal 9 g·L-1 saline group(n=20) and sites injected with VitB1 and VitB12 group(n=20).Rat models of HIE were prepared by ligation of left common carotid artery and later with a temporary systemic hypoxia for 2 hours.Fourteen days later,sites injected with 9 g·L-1 saline group and the sites injected with VitB1 and VitB12 group were respectively injected with 9 g·L-1 saline and mixed diluent VitB1 and VitB12 in the left frontal lobes and the apieal lobe of the rats one time a day for 20 days.Sham operation group and the model group were not treated.After 7 days intervention,rats in each group were killed,brains were removed after cardiac perfusion organizations with 40 g·L-1 paraformaldehyde,and a series of slices were made separately.The expression of Nestin used as neural stem cell marker in hippocampus was determined by immunofluorescence,and the expression of glial fibrillary acidic protein(GFAP) and neuron specific enolase(NSE) were detected with immunohistochemisty.The expression of nerve regene-ration inhibitory factor Nogo-A mRNA was determined by in situ hybridization.Results 1.Site medication injection could increase the expression of Nestin in hippocampus,with the highest expression when injected with VitB1 and VitB12,but with the lowest expression in sham operation group.2.In sham operation group and sites injected with VitB1 and VitB12 group,NSE expressions were higher,while the model group had the least expression(P0.05).3.The expressions of GFAP were least in hippocampus respectively in the sham operation and most in the model group by immunohistochemistry.Sites injected with 9 g·L-1 saline and sites injected with VitB1 and VitB12 could inhibit its expression,which were obviously for sites injected with VitB1 and VitB12 group(P0.05).4.The expression of Nogo-A mRNA was the least in the sham operation group,but the expression in the model group was in the opposite way.Sites injected with 9 g·L-1 saline group and sites injected with VitB1 and VitB12 group had a lower expression than that in the model group,which was obviously for sites injected with VitB1 and VitB12(P0.05).Conclusions Scalp site medication injection may stimulate endogenous neural stem cells proliferation and promote their differentiation,and also can inhibit expression of the nerve regene-ration inhibitory factor Nogo-A in brain,thereby promoting the repair of brain.

Objective To explore the clinical features,EEG characteristicsand prognosis of childhood frontal lobe epilepsy(FLE) in order to improve the cognition of FLE.Methods Twenty-four children with FLE collected from Jun.2009 to Jun.2010 in the neurology outpatient of the Third Affiliated Hospital of Zhengzhou University were reviewed retrospectively.All of them were examined by skull imaging studies and monitored by 24 h vigilance-controlled electroencephalogram(VEEG).Then the epileptic seizure types and seizure features,clinical characteristics,manifestations of EEG and iconography,therapeutic effects of antiepileptic drugs were analyzed.Results The seizures types were as follows:seizures of tic like(6 cases,25.00%),seizures of abnormal behaviors like(5 cases,20.83%),hypermotors(5 cases,20.83%),automatisms(3 cases,12.50%),torsive seizures(2 cases,8.33%),clonic seizures(2 cases,8.33%),asymmetric tonic seizures(1 case,4.17%).FLE typically came in short bursts that last less than 1 minute and often occur frequently while sleeping.All children had abnormal amount of discharge area,mainly for the amount of area origin of sharp waves,sharp slow wave,spike wave,spike slow wave,multi-spike slow wave,slow wave or more mixed,affecting central,and temporal region even all the leads.Five cases of them had imaging abnormality.After treatment of oral anti-epileptics(AEDs),21 cases(87.5%) were seizure free,and 3 cases(12.5%) got better.Conclusions Manifestations of seizures with FLE are various.The main types of seizures are tic like,abnormal behavior like,hypermotor automatisms,et al.Its features include short duration,frequent attacks and nocturnal preponderance.Routine EEG has low positive rate,so FLE has high misdiagnosis rate.The prolonged VEEG monitoring can increase the positive rate.The therapeutic effect of oral AEDs is good.