Cornuside alleviates experimental autoimmune encephalomyelitis by inhibiting Th17 cell infiltration into the central nervous system / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The present study was conducted to clarify the therapeutic effect of cornuside on experimental autoimmune encephalomyelitis (EAE) and its influence on T helper 17 (Th17) cell and regulatory T (Treg) cell infiltration into the central nervous system. Rats were randomly placed into four treatment groups: control, EAE, EAE+cornuside, and EAE+prednisolone. The neurological function scores of rats were assessed daily. On the second day after EAE rats began to show neurological deficit symptoms, the four groups were treated with normal saline, normal saline, cornuside (150 mg/kg), and prednisolone (5 mg/kg), respectively. The treatment was discontinued after two weeks, and the spinal cord was obtained for hematoxylin and eosin (H&E) and luxol fast blue staining, as well as retinoic acid receptor-related orphan receptor γ (RORγ) and forkhead box protein P3 (Foxp3) immunohistochemical staining. Blood was collected for Th17 and Treg cell flow cytometry testing, and the serum levels of interleukin (IL)-17A, IL-10, transforming growth factor-β (TGF-β), IL-6, IL-23, and IL-2 were measured via enzyme-linked immunosorbent assay (ELISA). Compared with rats in the EAE group, rats in the EAE+cornuside and EAE+prednisolone groups began to recover from neurological deficits earlier, and had a greater degree of improvement of symptoms. Focal inflammation, demyelination, and RORγ-positive cell infiltration were reduced by cornuside or prednisolone treatment, whereas the Foxp3-positive cell numbers were not significantly different. Meanwhile, the number of Th17 cells and the IL-17A, IL-6, and IL-23 levels were lower in the blood after cornuside or prednisolone treatment, whereas the number of Treg cells or the levels of IL-10, TGF-β, and IL-2 were not markedly different. Cornuside can alleviate symptoms of EAE neurological deficits through its anti-inflammatory and immunosuppressive effects, and Th17 cells may be one of its therapeutic targets.

Application of flipped classroom based on Wechat platform in the neurology teaching / 中华医学教育探索杂志

Objective:To discuss the application and effectiveness of the flipped classroom based on Wechat platform in the teaching of neurology.Methods:clinical students of grade 2014 of binjiang college of Zhejiang Chinese Medical University were selected as teaching subject. The students were divided into experimental group which contained a total of 74 students in class one and class three and the control group which contained a total of 38 students in class two. Peripheral neuropathy and neuromuscular-junction disease were selected as teaching contents. The experimental group adopted flipped classroom as the teaching mode and Wechat platform for pre-class preparation, after-school review and interactive communication. The control group was taught by traditional teaching mode. Before and after class, students in the two groups had a small-scale test and were surveyed by questionnaire, respectively. Students in the two groups took the same final exam which included case analysis when the course was over. The scores of the final exam and the results of the case analysis of each group were recorded and analyzed. All data were processed with statistical software SPSS 20.0, and t-test or chi-square test was used. Results:Students in the experimental group had significantly higher test scores in the after-class small-scale test than those in the control group ( P=0.038). Their final exam scores were higher than those in the control group ( P=0.046), and their scores of case analysis in the final exam were higher than those in the control group ( P=0.026). The results of pre-class questionnaire survey showed that the proportion of students who chose "good" in the experimental group was higher than that in the control group on the understanding of the learning content and the preparation ( P<0.05). In the after-class questionnaire survey, the proportion of students who chose "excellent" and "good" in the evaluation of learning interest in the experimental group was higher than those in the control group ( P<0.05), the proportion of students in the experimental chose "good" in evaluating their self-learning ability was higher than that in the control group ( P<0.05), the proportion of "excellent" on clinical thinking ability and teaching satisfaction was higher in the experimental group than that in the control group ( P<0.05), and the overall proportion of students who chose "excellent" and "good" in all items in the experimental group were higher than that in the control group ( P<0.05). In the experimental group, the overall proportion of students who selected "excellent" and "good" in on the evaluation of their learning interest was significantly increased in the after-class questionnaire survey compared with the pre-class questionnaire survey ( P<0.01). Conclusions:The application of flipped classroom based on Wechat platform is feasible and effective in the teaching of neurology. It can make up for the deficiency of traditional teaching methods. It is helpful to improve students' learning interest and self-learning ability, and is also helpful to exercise their clinical thinking ability. Thus this method deserves further popularization.

Protective effect of curcumin on dopamine neurons in Parkinson's disease and its mechanism / 浙江大学学报·医学版

OBJECTIVE@#To investigate the effect of curcumin on dopamine neurons in Parkinson's disease (PD) and its mechanism.@*METHODS@#SH-SY5Y human neuroblastoma cells were treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to establish the PD cell model. The model cells were treated with curcumin and/or autophagy inhibitor 3-MA. After 48 h of drug treatment, the number of surviving dopamine neurons was detected by tyrosine hydroxylase immunofluorescence method. Western blotting was used to detect protein expression of α-Synuclein (α-Syn), transcription factor EB (TFEB) and autophagy-related proteins lysosome-associated membrane protein 2A (LAMP2A) and microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ); RT-PCR was used to detect mRNA expression of α-Syn.@*RESULTS@#Compared with MPTP model group, curcumin increased the number of surviving dopamine neurons(<0.01), decreased both protein expression and mRNA expression of α-Syn (all <0.01), and increased protein expression of TFEB, LAMP2A and LC3-Ⅱ (all <0.01). When curcumin and 3-MA were given concurrently, the number of surviving dopamine neurons, protein expression of TFEB, LAMP2A and LC3-Ⅱ increased (<0.05 or <0.01), and both protein expression and mRNA expression of α-Syn decreased (<0.05 or <0.01) compared with MPTP model group; but the number of surviving dopamine neurons and protein expression of LAMP2A and LC3-Ⅱ decreased compared with curcumin group (all <0.05).@*CONCLUSIONS@#Curcumin exerts protective effect on dopamine neurons in PD, which may be associated with enhancing autophagy and promoting the clearance of α-Syn.

Effects of exercise on the expression of the LINGO-1 gene in rats after intra-cerebral hemorrhage / 中华物理医学与康复杂志

Objective To explore the effect of exercise on the expression of the LINGO-1 gene after intra cerebral hemorrhage (ICH).Methods Seventy-two Sprague-Dawley rats were randomly divided into a sham operation group,a control group and an exercise training group,each of 24.The three groups were further divided into 1 d,3 d,7 d and 14 d groups.ICH was induced using intra-parenchymal injection of autologous blood.The exercise training group was then forced to exercise on a treadmill.Any recovery of neurological functional was evaluated using Longa scoring,and the expression of LINGO-1 mRNA and protein were detected using the reverse transcription-polymerase chain reaction and western blotting.Results The sham operation group displayed no obvious neurological deficiency,with little expression of LINGO-1 mRNA or protein at any time point.The control group's average Longa score reached a maximum of 3 seven days after the operation,decreasing to 2 after another 7 days.The average expression of LINGO-1 mRNA and protein in that group peaked at 1.335±0.393 three days after the ICH,then decreased to 0.429±0.035 on the 7th day and 0.371±0.038 on the 14th day.In the exercise training group the average Longa score on the 7th day was 2,the average LINGO-1 mRNA level was 0.257±0.042 and the average protein level was 1.142±0.287,all significantly lower than in the control group.Moreover,in the exercise group there was a significant positive correlation between LINGO-1 protein expression and the Longa scores.Conclusion Exercise can decrease the expression of LINGO-1 mRNA and protein and promote the recovery of neural function after ICH.

Effects of exercise on the expression of the LINGO-1 gene in rats after intra-cerebral hemorrhage / 中华物理医学与康复杂志

Objective To explore the effect of exercise on the expression of the LINGO-1 gene after intra cerebral hemorrhage (ICH).Methods Seventy-two Sprague-Dawley rats were randomly divided into a sham operation group,a control group and an exercise training group,each of 24.The three groups were further divided into 1 d,3 d,7 d and 14 d groups.ICH was induced using intra-parenchymal injection of autologous blood.The exercise training group was then forced to exercise on a treadmill.Any recovery of neurological functional was evaluated using Longa scoring,and the expression of LINGO-1 mRNA and protein were detected using the reverse transcription-polymerase chain reaction and western blotting.Results The sham operation group displayed no obvious neurological deficiency,with little expression of LINGO-1 mRNA or protein at any time point.The control group's average Longa score reached a maximum of 3 seven days after the operation,decreasing to 2 after another 7 days.The average expression of LINGO-1 mRNA and protein in that group peaked at 1.335±0.393 three days after the ICH,then decreased to 0.429±0.035 on the 7th day and 0.371±0.038 on the 14th day.In the exercise training group the average Longa score on the 7th day was 2,the average LINGO-1 mRNA level was 0.257±0.042 and the average protein level was 1.142±0.287,all significantly lower than in the control group.Moreover,in the exercise group there was a significant positive correlation between LINGO-1 protein expression and the Longa scores.Conclusion Exercise can decrease the expression of LINGO-1 mRNA and protein and promote the recovery of neural function after ICH.

Effects of rapamycin and 3-MA on motor behavior and autophagy related protein LC3 in PD model mice / 中华行为医学与脑科学杂志

Objective To investigate the effects of rapamycin which is an autophagy inducer and 3-adenine (3-MA) which is an autophagy inhibitor on motor behavior and autophagy related protein LC3 in C57BL/6 mice of Parkinson's disease(PD).Methods 40 C57BL/6 male mice were randomly divided into control group and experimental groups which consist of MPTP model group,rapamycin group and 3-MA group,with 10 in each group.Mice in experimental groups received intraperitoneal injection with MPTP to establish PD models,and mice in control group received intraperitoneal injection with the same amount of saline solution for 1 week.Mice in rapamycin group received intraperitoneal injection with rapamycin and mice in 3-MA group received intraperitoneal injection with 3-MA at the same time when MPTP was injected.Animal behavior detections were carried out on the 1 th,7th and 14th day after the last injection.After the last injection,mice were sacrificed and sections of midbrain nigra were gained for the detection of expression of autophagy specificity protein LC3 by Western Blot.Results Compared with MPTP model group,rapamycin could improve the general condition and behavioral manifestation of mice in pole test((14.89± 1.14) s vs (16.24±1.39) s,P＜0.05;(15.18±1.36) s vs(17.93±1.11s),P＜0.01),traction test((1.7±0.5) vs (1.2±0.4),P＜ 0.05;(1.5±0.5)vs (1.1±0.3),P＜0.05) and open field test which contained total activity distance((5 875.3 ± 1148.9) cm vs (5 061.5±773.1) cm,P＜0.05;(5 753.2± 1 106.7) cm vs (4 669.3±969.1) cm,P＜0.01) and average speed((19.29±2.35) cm/s vs (16.47±3.01) cm/s,P＜0.05;(18.71±2.71)cm/s vs (15.80±2.50) cm/s,P＜0.01),while 3-MA aggravated behavioral deficits of mice in pole test(19.92± 1.61s vs 17.93± 1.11 s,P＜0.05),and both total activity distance((3 879.7±575.0) cm vs (4 669.3±969.1) cm,P＜0.05) and average speed((13.34± 1.87) cm/s vs (15.80±2.50) cm/s,P＜0.05) in open field test were decreased.The results of Western Blot confirmed that rapamycin could increase the expression of LC3-Ⅱ,however 3-MA could re duce the expression of LC3-Ⅱ.Conclusion This study confirmed that rapamycin could alleviate behavioral symptoms of PD model mice and increase the level of LC3 in midbrain nigra,while 3-MA could exacerbate behavioral symptoms in PD model mice and decrease the level of LC3 in midbrain nigra.Thus it can be speculated that drugs which can regulate autophagy may be potential treatment protocols for PD.