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.

Objective:To clarify the infection and epidemic characteristics of the human metapneumovirus (HMPV) in Chinese patients with febrile respiratory syndrome (FRS), and to provide important baseline data for clinical diagnosis, treatment, prevention and control of HMPV-induced respiratory tract diseases in China.Methods:FRS cases from January 2009 to June 2021 in 9 provinces in China, including Beijing, Hebei, Jilin, Shandong, Shaanxi, Xinjiang, Anhui, Guangdong, Hunan were retrospectively analyzed for their respiratory samples, clinical and epidemic data.The respiratory samples were detected for HMPV by quantitative real-time PCR.Results:A total of 11 660 cases were tested for HMPV, involving 296 (2.54%) HMPV-positive cases.Among 296 HMPV-positive cases, 218 were single HMPV infection, and 78/296 (26.35%) were co-infected with one or more respiratory viruses.HMPV mainly affected children under 5 years of age (3.10%), and in this population, the proportion of pneumonia in HMPV co-infection cases was significantly higher than that of single HMPV infection.HMPV could be detected all year round, which was more popular in winter and spring, with the peak of HMPV epidemic in March.Conclusions:HMPV is one of the important pathogens causing acute respiratory infection in children, showing a clear seasonal epidemic.HMPV can be infected alone or in combination with other respiratory viruses, which may increase the risk of pneumonia in children.

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.

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.
Animals ; Cell Line ; Curcumin ; pharmacology ; Dopaminergic Neurons ; drug effects ; Humans ; Mice ; Mice, Inbred C57BL ; Parkinson Disease ; alpha-Synuclein ; metabolism

Objective To investigate whether curcumin protects dopamine neurons in Parkinson's disease (PD) by activating autophagy and discuss its specific mechanism.Methods In vitro,SH-SY5Y human neuroblastoma cells were treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPP+) to induce PD cell models.In vivo,C57BL/6 male mice were performed intraperitoneal injection of 30 mg/kg 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) once daily for 7 d continuously to establish PD models.SH-SY5Y human neuroblastoma cells and C57BL/6 male mice were,respectively,divided into normal control group,model group,curcumin group,curcumin and 3-methyladenine (3-MA) group,and 3-MA group.Curcumin was used as intervention treatment,and 3-MA,an autophagy inhibitor,was used individually or concurrently with curcumin as intervention controls:curcumin (40 μmol/L),curcumin (40 μmol/L)+3-MA (4 mmol/L),and 3-MA (4 mmol/L) were added into the cells of curcumin group,curcumin and 3-MA group,and 3-MA group at the time of MPP+ addition;curcumin (80 mg/kg),curcumin (80 mg/kg)+3-MA (2 mg/kg),and 3-MA (2 mg/kg) were added into the mice of curcumin group,curcumin and 3-MA group,and 3-MA group at the time of MPTP injection once daily.In the normal control group,sterile saline was injected into the abdomen simultaneously (30 rmL/kg).In cell groups,after 48 h of drug treatment,the number of surviving dopamine neurons was detected by tyrosine hydroxylase (TH) immunofluorescence;Western blotting was used to detect protein expressions of α-synuclein (α-Syn),and autophagy related proteins lysosome-associated membrane protein 2A (LAMP2A) and microtubule-associated protein 1 light chain 3 (LC3);transcription factor EB (TFEB) levels in the cytoplasm and nucleus were detected separately by Western blotting;reverse transcription (RT)-PCR was used to detect the α-Syn mRNA expression.In mice groups,14 d after the last injection,mice were sacrificed and sections ofmidbrain nigra were gained for detection of TH level by immunohistochemistry,expressions of α-Syn and LC3 were detected by Western blotting.Results (1) As compared with model group,curcumin group had significantly larger number of surviving dopamine neurons both in vitro and in vivo,significantly decreased α-Syn protein and mRNA expressions in vitro,and significantly decreased α-Syn protein expression in vivo (P＜0.05);curcumin group had statistically increased LAMP2A and LC3-Ⅱ protein expressions in vitro and LC3-Ⅱ protein expression in vivo as compared with model group (P＜0.05).(2) As compared with curcumin group,curcumin and 3-MA group had statistically smaller number of surviving dopamine neurons in vitro and in vivo,and statistically increased α-Syn protein expression in vitro (P＜0.05).(3) In vitro,as compared with model group,curcumin group had significantly increased TFEB protein expression in nucleus (P＜0.05),and curcumin and 3-MA group had statistically increased TFEB protein expression in cytoplasm and statistically decreased TFEB protein expression in nucleus (P＜0.05).Conclusion Curcumin has protective effect on dopamine neurons of PD;one of the main mechanisms is that curcumin can activate the autophagy function and promote the clearance of α-Syn;increasing the TFEB expression,promoting the transfer of TFEB to nucleus to perform the transcription function,and then promoting the synthesis of autolysosome may be the main ways to activate the autophagy function.

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.

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.

Objective To investigate the effect of cystatin C (Cys C) on adventitia in rabbit abdominal aorta restenosis after angioplasty and its mechanism.Methods 48 New Zealand white rabbits were randomly divided into injury group (receiving balloon dilation of abdominal aorta),the treatment group (taking Cys C monoclonal antibody therapy) and the control group (receiving femoral artery puncture and catheter sheath without balloon dilation and intervention of Cys C monoclonal antibody injection),and each group had 16 rabbits.Peripheral vein blood was drawn to measure the serum level of cystatin C before and 8 h,1 day,1 week,3 weeks,6 weeks after the operation in all rabbits.After 6 weeks of operation,the abdominal aorta were taken and stained with HE.Vascular morphometry analysis and adventitial cell count were conducted.Smooth muscle actin (SM-actin) and proliferating cell nuclear antigen (PCNA) expressions in the adventitia were observed by immunohistochemical staining.The number of PCNA positive cell in the adventitia was counted and the PCNA proliferation index was calculated.The vascular remodeling index,vascular external elastic lamina area (EELA),internal elastic lamina area (IEIA) were used to evaluate the vascular remodeling and the residual stenosis and vascular cavity area was used to measure the vascular stenosis.Results Plasma Cys C level began to rise at 8h after operation and reached the peak at 1 week after operation,and continuously increased for 5 weeks in injury group,and reached to respectively at 3 weeks and 6 weeks after operation.The Cys C levels were significantly higher in injury group than in the treatment and control groups at different time points (all P＜0.05).There were no significant differences in Cys C levels at different time points between the treatment group and the control group.The injury group showed that the number of PCNA positive cells was higher in injury group than in treatment and control groups,both P＜0.05).Compared with the control group,the vascular luminal area,EELA and IELA were significantly increased (all P＜0.05).After treated with the monoclonal antibody Cys C intervention,the treatment group showed that lumen area,vascular EELA,IELA was significantly decreased (P＜0.05),and the vascular remodeling index and residual stenosis rate were decreased as compared with the injury group (0.871 vs.0.784,33.1％ vs.19.7 ％,both P＜0.05).Correlation analysis showed that Cystatin C level was positively correlated with the vascular lumen area,neointimal area,internal elastic lamina area,external elastic lamina area and the number of PCNA positive cells (r=0.812,0.797,0.876,0.932 and 0.822 respectively,all P＜0.01).Conclusions Plasma Cys C level is increased in rabbit after abdominal aorta balloon injury and has a positive correlation with the severity of arterial stenosis.High Cys C level can induce adventitial fibroblast activation,proliferation,phenotype transformation and migration,and accelerate the processes of atherosclerosis and stenosis.Cys C level is the independent risk factor for abdominal aortic stenosis.

Objective:To investigate the clinical efficacy of posterolateral locking plate in the treatment of large posterior malle-olar fracture .Methods :From January 2010 to June 2012 ,23 patients with large posterior malleolar fractures were treated by posterolateral locking plate ,including 15 males and 8 females ,all of whom aged from 32 to 78 years with an average age of 43 .7 years .The average time from injury to operation was 7 .3 days ,which ranged from 5 to 12 days .Calcaneal traction fixa-tion or plaster was performed on the injured limb before operation .Once that swelling reduced ,all patients were treated by posterolateral locking plate and were assessed with the ankle hindfoot clinical rating system of the American Orthopaedic Foot and Ankle Society (AOFAS) .Results:The follow-up time was from 14 to 22 months ,with an average period of 17 .4 months . 23 patients resulted in bone healing ,without loosen or breakage of internal fixation .As the X-ray pictures showed ,fracture healing time after operation ranged from 11 to 21 weeks ,which was 14 .7 weeks on average ,and full weight bearing time was from 15 to 25 weeks ,which was 17 .1 weeks on average .The AOFAS scores ranged from 73 to 94 points with an average of 83 .4 points at the 12th month after operation .One patient had superficial wound infection at the 7th day after operation and was healed by sensitive antibiotics and changing fresh dressing .Conclusions :Posterolateral locking plate could be a good method for the treatment of posterior malleolar fracture because effective redution and proper fixation could be obtained through straight access .

Objective To investigate the effects of the treatment of open injury and orthopedic postoperative wound infections with Vacuum Sealing Drainage.Methods Retrospective analysis of the clinical results of 32 patients with open injury or orthopedic postoperative wound infections in our hospital.Results Follow the indications and contraindications strictly and ensure right pressure,all the infection of wounds was controlled,and split skin graft survived.Without the package and pressurized fixation,the wounds healed better.Conclusion Vacuum Sealing Drainage can be considered one of the effective methods to treat the open injury or orthopedic postoperative wound infections and is worthy to be carried out as a routine.For patients with vascular rupture or defect,when there was still skin defects after vascular graft repair,as long as covered vascular anastomosis with the surrounding skin and soft tissue during the operation,there was still can use closed vacuum sealing drainage to cover the wound to prevent infection.