Objective To investigate expression of glycogen synthase kinase(GSK)-3? and phosphorylation GSK-3? (p-GSK-3?) in the anterior temporal neocortex in patients with refractory epilepsy(RE). Methods Expression of GSK-3? and p-GSK-3? were detected by RT-PCR, FQ-PCR, Western Blot and immunohistochemistry in the anterior temporal cortices of 36 RE cases. 8 patients without RE had been used as the controls. Results Compared with control group,the expression of GSK-3? mRNA and protein were significant higher(allP

Objective To establish a high-performance liquid chromatography (HPLC) method with diode array detection to simultaneously determine carbamazepine,phenytoin,and phenobarbital in serum.Methods Extraction solvent (800μl ethylene acetate) and sample (0.2 ml) was mixed,extracted for 2 min,and centrifuged (3500 r/min,4 minutes).A volume (600 μl) of extract liquor was volatilized to dryness in water bath with the volatilization temperature 75 ℃,then was redissolved with 1.0 ml mobile phase.Analysis conditions was column temperature 30°,mobile phase (methanol∶ water =40∶60),and detection wavelength of 254 nm.Three metabolites were effectively separated.Results Under the optimized condition,calibration curves of three metabolites were linear in the ranges of (1.52 ～ 120 mg/L) and the correlation coefficients were not less than 0.999.The detection limits (S/N =3) were in the range of 0.4 ～ 1.5 mg/L.The spiked recoveries were in the range of 91.3％ ～ 111％ with relative standard deviations (RSD) less than 5％.Conclusions The optimal pretreatment condition for the sample was established.The chromatographic separation and the detection condition were optimized.The method was sensitive and accurate,and could meet the need of monitoring serum drug concentration.

Objective To study the validity and reliability of Tengdao's swallowing standard for stroke pa-tients with dysphagia.Methods A total of 128 patients with postroke dysphagia took the swallowing test and then were divided into three sub-groups.Their scores on Tengdao's evaluation and their fluoroscopy results were analyzed using Spearman's correlation coefficient.Intra-class coefficients (ICCs)were used to examine the intra-rater and in-ter-rater reliability of Tengdao's evaluation.Results TenIgdao's evaluation possessed good validity and reliability.There was a high correlation between the scores in Tengdao's evaluation and fluoroscopy results. Conclusions Tengdao's evaluation is valid,reliable,simple and safe.It can be used in the clinic to evaluate the stroke patients with dysphagia.

Objective To investigate the effect of Shenxiong glucose injection on platelet reactivity during aspirin treatment in patients with acute ischemic stroke.Methods A total of 263 patients with acute ischemic stroke admitted to 12 hospitals from January 2014 to December 2016 were enrolled prospectively.They were randomly divided into aspirin group and aspirin + Shenxiong glucose injection group.The changes of platelet maximum aggregation rate induced by 4 platelet aggregating agents (arachidonic acid,adenosine diphosphate,collagen and platelet activating factor) were detected before and after the treatment.Results There were no significant differences in the demographic data and baseline clinical characteristics between the aspirin group (n =132) and the Shenxiong glucose injection + aspirin group (n =131).At baseline,the maximum aggregation rate of platelet induced by arachidonic acid and platelet activating factor in Shenxiong glucose injection + aspirin group was significantly higher than that in the aspirin group (all P ＜0.05).On the 6th day after treatment,the maximum aggregation rate of platelets induced by the 4 aggregating agents in the Shenxiong glucose injection + aspirin group was significantly lower than that in the aspirin group (all P ＜ 0.001).Conclusion Shenxiong glucose injection had a significant inhibitory effect on platelet reactivity during aspkin treatment in patients with acute ischemic stroke.

Objective To investigate whether GLT25D2 gene regulates autophagy in acetaminophen (APAP)-induced hepatotoxicity injury.Methods GLT25D2+/+ wild-type C57BL/6J mice and GLT25D2-/- C57BL/6J mice were selected as subjects. ①In vivo experiment: 20 for wild-type mice and 20 for GLT25D2-/- mice were respectively divided into phosphate buffer (PBS) control group and APAP intervention group according to random number table, with 10 mice in each group. The hepatotoxicity injury model of mice was reproduced by intraperitoneal injection of 25 g/L APAP solution 500 mg/kg. The PBS control group was intraperitoneally injected with the same amount of PBS. The mice were sacrificed immediately after model reproduction, and the liver tissues were harvested. Western Blot was used to detect the expressions of autophagy-related proteins ATG5, ATG7, microtubule-associated protein 1 light chain 3 (LC3) and P62. The ultrastructural changes in liver tissue were observed under electron microscope to observe the level of autophagy. ②In vitro experiment: primary hepatocytes extracted by two-step collagen perfusion from one GLT25D2+/+wild-type mouse and one GLT25D2-/- mouse were divided into two parts respectively. One part was treated with 5 mmol/L APAP solution. The cells were harvested at 0, 8, and 12 hours, and the expressions of autophagy-related proteins ATG5, ATG7, LC3, and P62 were determined by Western Blot. The other part was transfected with the green fluorescent protein-LC3 plasmid (GFP-LC3) for 24 hours. The cells were cultured with PBS (PBS control group) or 5 mmol/L APAP (APAP intervention group) for 12 hours, and the positive expression of GFP-LC3 was observed under the fluorescence microscope, thereby reflecting the expression of autophagosomes.Results ①In vivo experiment: compared with the corresponding PBS control group, the expressions of the positive-associated proteins ATG5, ATG7 and LC3-Ⅱ in liver tissue of the APAPintervention group were down-regulated in the wild-type and GLT25D2-/- mice, while the expression of the negative correlation protein P62 was up-regulated, indicating that the overall level of autophagy decreased after treatment with APAP. Compared with wild-type mice, the expressions of autophagy positive correlation proteins ATG5 and ATG7 were up-regulated in GLT25D2-/- mice (ATG5/β-actin: 1.21±0.29 vs. 0.84±0.19, ATG7/β-actin:1.29±0.14 vs. 1.54±0.40, bothP > 0.05), LC3-Ⅱ expression was slightly down-regulated (LC3-Ⅱ/β-actin: 0.52±0.06 vs. 0.58±0.06,P > 0.05), while negative correlation protein P62 was down-regulated (P62/β-actin: 1.13±0.94 vs. 1.54±0.40,P > 0.05), indicating that the expression of autophagy in GLT25D2-/- mice was higher than that in wild-type mice. Ultrastructural observation under electron microscope showed that the number of autophagosomes in the liver tissue of wild-type mice did not change significantly after APAP intervention as compared with that in PBS control group, but the number of autophagosomes in GLT25D2-/- mice was increased. ②In vitro experiment: with the prolongation of APAP intervention, the expressions of ATG5 and ATG7 in the primary hepatocytes of wild-type and GLT25D2-/-mice were up-regulated, LC3 was slightly fluctuated, and the expression of negative-related protein P62 was gradually down-regulated. The peak value or the trough value reached at 12 hours. It was indicated that the expression of autophagy in APAP-stimulated cells was enhanced with a time-dependent manner. Compared with wild-type mice, the expressions of autophagy correlation proteins ATG5, ATG7, LC3-Ⅱ and P62 were up-regulated in GLT25D2-/- mice at 12 hours (ATG5/β-actin: 0.93±0.09 vs. 0.74±0.06, ATG7/β-actin: 0.80±0.09 vs. 0.65±0.10, LC3-Ⅱ/β-actin:1.35±0.30 vs. 1.15±0.20, P62/β-actin: 0.36±0.02 vs. 0.31±0.03, allP > 0.05), indicating that the expression of autophagy was enhanced after gene knockout. Fluorescence microscopy showed that GFP-LC3 positive cells in both wild-type and GLT25D2-/- mice hepatocytes were significantly increased after APAP intervention as compared with those of PBS control group, and the proportion of GFP-LC3 positive cells in GLT25D2-/- mice was significantly higher than that in wild-type mice (0.64±0.08 vs. 0.36±0.05,P < 0.05).Conclusions GLT25D2 is a negative regulator of autophagy. Knockout of GLT25D2 gene can enhance the autophagy level of APAP-induced hepatotoxicity injury in mice.

Objective To investigate expression of extracelluar signal regulated protein kinase(ERK)and phosphorylation ERK(p-ERK)in the temporal lobe from patients with DR-TLE so as to explore the possible roles of ERK in the pathogenesis of DR-TLE.Methods Expression of ERK was detected with Western blot and immunohistochemistry in 32 patients with DR-TLE(24 temporal lobe,8 hippocampi),as compared with 12 controls(9 temporal lobe,3 hippocampi).Results ERK and p-ERK expression in DR-TLE was significantly higher(0.2266?0.0613,0.2097?0.0183 and 0.1924?0.0054,respectively)than those of controls(0.1840?0.0023,0.1974?0.0056 and 0.1825?0.0063,respectively,all P

Objective To explore the effect of high-intensity ultrasound focused ablation in the treatment of ⅡB cesarean scar Pregnancy.Methods A total of 365 patients in our hospital from January 2019 to December 2021 which were ⅡB Cesarean Scar Pregnancy were selected as the research objects.The research objects were divided into the experimental group pretreated by high-intensity ultrasound ablation(186 cases)and the control group without treatment(179 cases)to compare the levels of intraoperative blood loss,postoperative vaginal bleeding time,human chorionic gonadotropin(HCG)decline rate after the operation,success rate of operation and other indicators.Results The amount of intraoperative blood loss and postoperative vaginal bleeding time in the experimental group were less than those in the control group(P<0.05),and the decrease rate of blood HCG in the experimental group was higher than that in the control group(P<0.05).There was significant difference in the success rate of operation between the experimental group and the control group(P<0.05).There was no significant difference in menstrual volume between the experimental group and the control group(P>0.05),but the control group had prolonged menstruation,and the incidence of postoperative complications between the experimental group and the control group have no statistical significance.Conclusion High-intensity focused ultrasound ablation can effectively reduce intraoperative blood loss in patients of type ⅡB cesarean scar pregnancy,reduce the application of invasive methods such as laparoscopy,can be promoted as a pretreatment method for type ⅡB cesarean scar pregnancy.