Objective:To explore the correlation of serum neurogenic exosome MicroRNA-211-5p(miR-211-5p)levels and brain derived neurotrophic factor(BDNF)levels with cognitive impairment in patients with Parkinson's disease and their diagnostic value.Methods:A total of 80 patients with Parkinson's disease(PD)admitted to the Second Hospital of Jiaxing City from January 2017 to April 2018 were enrolled.According to the Montreal cognitive assessment scale, patients were divided into the cognitive impairment group(n=36)and the non-cognitive impairment group(n=44). Meanwhile, 30 healthy people who took health check-ups during the same period were selected as the control group.Exosomes were extracted from peripheral blood of subjects by using the ExoQuick kit, and the neurogenic exosomes were separated by an L1 cell adhesion molecule(L1CAM)biotinylated antibody.BDNF levels in the exosomes were measured with an enzyme-linked immunosorbent assay(ELISA), and the expression level of miR-211-5p in the exosome was determined by real-time fluorescence quantitative PCR(RT-QPCR).Results:There was a correlation between BDNF and miR-211-5p( r=-0.805, P<0.001)in serum neurogenic exosomes( r=-0.805, P<0.001). BDNF was correlated with miR-211-5p in both the PD and control groups( r=-0.785 and-0.867, P=0.002 and 0.001). The miR-211-5p level was higher and the BDNF level was lower in the PD group than in the control group(0.30±0.08 vs. 0.17±0.04, 0.55±0.06 mg/L vs. 0.75±0.06 mg/L, t=7.125 and 6.368, P=0.000 and 0.000). The BDNF level was lower(0.45±0.07 mg/L vs.0.63±0.07 6.368 and 0.75±0.08 mg/L, t=8.999 and 7.608, P=0.000 and 0.000)and the MiR-211-5p level was higher(0.36±0.07 vs. 0.24±0.05 and 0.17±0.04, t=10.923 and 7.520, P=0.000 and 0.000)in the cognitive impairment group than in the non-cognitive impairment and control groups.The receiver-operating characteristics(ROC)curve showed that the area under the curve of miR-211-5p as a measure for cognitive impairment in Parkinson's disease was 0.860(95% CI: 0.770-0.950)with a threshold of 0.32.The area under the curve of BDNF as a measure for cognitive impairment in Parkinson's disease was 0.891(95% CI: 0.822-0.961)with a threshold of 0.67.BDNF seemed to be the target gene of miR-211-5p, since the latter could inhibit BDNF expression by reducing BDNF mRNA levels. Conclusions:Human serum neurogenic exosome miR-211-5p is highly expressed in PD patients with cognitive impairment and has the potential to be used as one of diagnostic parameters for cognitive impairment in PD patients.The high expression of serum neurogenic exosome miR-211-5p may be related to the inhibition of BDNF by reducing its mRNA levels.

Objective To study the effect of intestinal bacteria on motor ability of amyotrophic lateral sclerosis (ALS) mice models and its mechanism. Methods Twenty wild type C57BL/6J mice (WT group) and 20 SOD1-G93A transgenic ALS mice (ALS group) were selected as the research subjects. (1) Ten mice in both WT group and ALS group were selected, respectively; 5 mice in each group were fed in SPF environment, and the remaining 5 mice were fed in aseptic environment; they were defined as WT+SPF group, WT+aseptic group, ALS+SPF group and ALS+aseptic group. (2) Ten mice in WT group and ALS group were fed in sterile environment; 5 mice in each group were transplanted with fecal bacteria, and the remaining 5 mice were not interfered; they were defined as WT+transplantation group, WT+non-transplantation group, ALS+transplantation group and ALS+non transplantation group. The grip strength of mice was measured by grip force meter, the motor coordination ability of mice was tested by roller treadmill and rotating rod test, the number of motor neurons in the anterior horn of spinal cord was measured by Nissl staining, the expression of microglia activation marker ionic calcium junction protein (IBA-1) in spinal cord tissues was detected by immunohistochemical staining, and the expressions of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in spinal cord tissues were detected by Western blotting; the β-N-methylamino-L-alanine (BMAA) expression was detected by high performance liquid chromatography-tandem mass spectrometry. Results (1) The grip strength, drop latency and drop time of ALS+aseptic mice were significantly higher than those of ALS+SPF mice, the number of Nissl-stained positive cells was significantly larger than that of ALS+SPF mice, the number of IBA-1 positive cells was significantly smaller than that of ALS+SPF mice, the levels of TNF-α and IL-6 protein expressions and BMAA concentration were statistically lower than those of ALS+SPF mice (P<0.05). (2) The grip strength, drop latency and drop time of ALS+transplantion mice were significantly lower than those of ALS+non-transplantation mice, the number of Nissl-stained positive cells was significantly smaller than that of ALS+non-transplantation mice, the number of IBA-1 positive cells was significantly larger than that of ALS+non-transplantation mice, the TNF-α and IL-6 protein expressions and BMAA concentration were significantly higher than those of ALS+non-transplantation mice (P<0.05). Conclusion Imbalance of intestinal bacteria homeostasis can decrease the motor ability of ALS mice, which is related to the activation of microglia.

Objective To investigate the proportion of helper T cell subset Th 9 and the expression of its cytokine interleukin-9(IL-9)in Parkinson's disease(PD)and the clinical significance.Methods Seventy-two patients diagnosed with PD between January 2016 and June 2017 and 20 healthy volunteers in the same period were selected.The PD patients were staged according to the Hoehn-Yahr(H-Y)staging method,21 in stage one,19 in stage two,18 in stage three,11 in stage four,and three in stage five.The proportion of Th9 subset in peripheral blood of PD patients and healthy volunteers was measured by flow cytometry.The expression of IL-9 in peripheral blood of PD patients and healthy volunteers was measured by enzyme-linked immunosorbent assay.Results The proportion of Th9 cells in peripheral blood of PD patients(1.27%±0.34%)was significantly higher than that of healthy volunteers(0.61%±0.11%,t=8.530,P<0.05),and the higher stage of PD,the higher proportion of Th9,suggesting that the proportion of Th9 was related to the PD staging.IL-9 was also highly expressed in PD patients((16.04 ±2.94) pg/ml)and had statistically significant difference compared with healthy volunteers((7.53 ±0.70)pg/ml;t=12.781,P<0.05).IL-9 was similar to Th9, the higher stage of PD, the higher expression of IL-9. Conclusion The proportion of Th9 cells and the expression of IL-9 in peripheral blood of patients with PD increased significantly,having a significant relationship with the H-Y staging of PD.

Objective To investigate the effects of dl-3-n-butylphthalide ( NBP) on angiogenesis of human umbilical vein endothelial cells ( HUVEC) in vitro under ischemia/hypoxia and the correlation with vascular endothelial growth factor/vascular endothelial growth factor receptor 2 ( VEGF/VEGFR2 )-Notch1/Dll4 signaling pathway .Methods The control group , ischemia/hypoxia group , and ischemia/hypoxia +NBP group ( high dose group and low dose group ) were set up after HUVEC subculture . The cell concentration was adjusted to be 1 ×105/ml, and each group was randomly added 1 ml (1 ×105 cells in each group).Both of the ischemia/hypoxia group and the ischemia/hypoxia +NBP group were cultured under the condition of ischemia and hypoxia .The NBP concentrations of the high and low dose groups were 20 μmol/L and 5 μmol/L respectively.The cell viability of each group was detected by cell counting kit-8, and cell scratch test was used to detect the migration ability of the cells in each group .The cell formation ability of each group was examined by in vitro angiogenesis assay.Western blotting was used to detect the expressions of receptor proteins VEGFR2, Notch1 and Dll4,and mRNA expressions of VEGF, VEGFR2, Notch1 and Dll4 were detected by real-time quantitative PCR.Results NBP increased the survival rates of HUVEC under ischemia/hypoxia condition.In the low dose group, the survival rates of HUVEC at 6, 12, 24, 48 h were 78.6％ ±3.0％, 59.6％ ±5.3％, 44.6％ ±4.2％, 38.2％ ±4.3％, respectively, significantly higher than that in the ischemia/hypoxia group (75.2％±5.8％, 53.2％±4.8％, 36.2％± 7.8％, 22.5％±4.1％;t=4.513, 6.231, 9.322, 9.674; P=0.021, 0.018, 0.026, 0.015).In the high dose group, the survival rates of HUVEC at 6, 12, 24, 48 h were 88.6％±6.3％, 67.5％±5.4％, 53.3％±4.2％, 46.3％±3.9％, respectively , also significantly higher than that in the ischemia/hypoxia group (t=8.123, 11.211, 12.312, 14.154;P=0.001, 0.002, 0.001, 0.001).The mobility of the low dose group was 52.3％+4.2％, with statistically significant difference compared with that in the ischemia /hypoxia group ( 18.5％ ±3.2％) and the control group ( 22.3％ ±4.1％; t=18.324, 15.183; P=0.000, 0.000).The mobility of the high dose group was 87.5％±5.2％, also with statistically significant difference compared with that in the ischemia/hypoxia group and the control group ( t=22.142, 19.341;P=0.000, 0.000).NBP increased the protein and mRNA expression of VEGF , VEGFR2, Notch1 and Dll4.The relative expression of VEGFR2, Notch1 and Dll4 in the low dose group was 1.12 ±0.17, 0.35 ± 0.07 and 0.42 ±0.08, respectively, with statistically significant difference compared with that in the ischemia/hypoxia group (0.82 ±0.05, 0.30 ±0.03, 0.32 ±0.04;t=6.120, 2.123, 4.112;P=0.000, 0.020, 0.003).The relative expression of VEGFR2, Notch1 and Dll4 in the high dose group was 1.30 ± 0.15, 0.41 ±0.10 and 0.48 ±0.11, respectively, also with statistically significant difference compared with that in the ischemia/hypoxia group ( t=8.122, 3.851, 5.130; P=0.000, 0.000, 0.001 ) .The mRNA expressions of VEGF , VEGFR2, Notch1 and Dll4 in the low dose group were 0.43 ±0.08, 0.41 ± 0.05, 0.38 ±0.03 and 0.36 ±0.04, respectively, with statistically significant difference compared with that in the ischemia/hypoxia group (0.28 ±0.03, 0.34 ±0.04, 0.27 ±0.03, 0.19 ±0.04;t=3.122, 3.825, 4.311, 5.211; P=0.000, 0.006, 0.001, 0.000).And the mRNA expressions of VEGF, VEGFR2, Notch1 and Dll4 in the high dose group were 0.58 ±0.05, 0.50 ±0.06, 0.41 ±0.05, 0.52 ±0.06, respectively, also with statistically significant difference compared with that in the ischemia /hypoxia group (t=4.225, 4.872, 5.311, 8.220;P=0.000, 0.000, 0.000, 0.000).Conclusions NBP can promote HUVEC to form blood vessels under ischemia/hypoxia condition , the mechanism of which may be related to the activation of VEGF/VEGFR2-Notch1/Dll4 signaling pathway .It may be one of the mechanisms that NBP improves cerebral microcirculation in acute ischemic stroke .

Objective: To study the mechanism of interleukin (IL)-17 in mice with non-alcoholic fatty liver disease for promoting M1-type macrophage polarization to exacerbate liver inflammation, and to provide references for the mechanism of NAFLD occurrence and development. Methods: A mouse model of NAFLD was constructed by high-fat diet. Mice were divided into control group, model group, IL-17 group, and anti IL-17 group. Histopathological changes of the liver were observed by HE staining. The serum levels of ALT and AST in peripheral blood of mice was detected by chemical colorimetry. Macrophages labeled with F4/80-PE, CD11C-FITC was designated as M1-type macrophages, those labeled with F4/80-PE, and CD206-APC was designated as M2-type macrophages. The proportion of M1 and M2 macrophages infiltrated into the liver tissues of mice were measured by flow cytometry. CD168 expression level of liver tissues was detected using immunohistochemistry. Protein and mRNA levels of the marker molecules (iNOS, TNF-alpha and IL-6) of M1 macrophages were detected using ELISA and RT-Q PCR. Western blot was used to detect the protein expression of JAK-STAT signal pathway and the expression level of MCP-1. Data were analyzed using one-way ANOVA and t-test. Results: High-fat diet NAFLD mice model was successfully constructed. IL-17 had increased the proportion of M1 macrophages in mice liver tissues and decreased the proportion of M2 macrophages (P < 0.05). The proportion of M1 and M2 macrophages in the liver tissues of normal mice was 7.9% ± 1.1% and 19.2% ± 1.8%. The proportion of M1 and M2 macrophages in the model group was 17.3% ± 2.5% and 15.0% ± 2.1. The proportion of M1 macrophages (33.8% ± 4.2%) in IL-17 group was higher than model group, while the proportion of M2 macrophages (7.8% + 1.0%) in IL-17 group was lower than model group. Protein and mRNA marker levels of M1 macrophage (iNOS, IL-12, TNFα and IL-6) in liver tissues were significantly higher than model group, control group, and anti-IL-17 group (P < 0.05). The expression levels of JAK1, STAT1, MCP-1, and CD168 in mice liver tissues of IL-17 group had increased (P < 0.05). The levels of aspartate and alanine aminotransferases in peripheral blood of mice in IL-17 group were significantly higher than other three groups (P < 0.05). Conclusion IL-17 can promote M1-type macrophage polarization, and exacerbates the liver inflammatory response to accelerate the progression of NAFLD in mice.

Objective To explore the the characteristics of non-motor symptoms of essential tremor(ET).Methods Totally 50 ET patients and 45 age-gender-matched healthy volunteers,admitted in Department of Neurology,The Second Affiliated Hospital of Zhejiang University School of Medicine from May 2015 to April 2016,were included.Clinical data and tremor analyses under different postures were obtained.The non-motor symptoms were evaluated using the Pittsburgh Sleep Quality Index(PSQI),Hamilton anxiety scale(HAMA),Hamilton's Depression Scale(HAMD),and the MOS item short from health survey (SF-36).Cognitive functions were evaluated by the Minimental state examination(MMSE).Results ET group had lower MMSE total score of (25.81 ±2.75 vs.28.16 ± 1.71),increased rate of dyssomnia (62.0％ vs.15.6％) and higher PQSI score (6.42±2.71 vs.3.84±2.13)compared with the control group(all P＜0.05).Moreover,more patients in the ET group had moderate and severe anxiety(60.0 ％ vs.37.8 ％),moderate and severe depression (34.0％ vs.15.6％)than the control group(both P ＜ 0.05).Anxiety and depression had effects on physical and mental health and were also related to the quality of life.Conclusions Non-motor symptoms,such as mild cognitive deficits,depression,anxiety and dyssomnia are common in ET patients.Furthermore,depression and anxiety have negative effects on physical and mental health.

Objective To analyze the clinical features of Parkinson's disease (PD) combined with chronic pain in the elderly.Methods A total of 366 idiopathic PD patients experiencing pain were enrolled and divided into two groups:the elderly group (n=289) and the young group (n=77).Rating scales including Visual Analogue Scale (VAS),Unified Parkinson's Disease Rating Scale (UPDRS),Hoehn Yahr (H-Y) Scale,Self-rating Anxiety Scale (SAS),Self rating Depression Scale (SDS) and Brief Pain Inventory (BPI) were evaluated.Results Compared with the young group,the elderly group had evidently higher scores of UPDRSⅡ,Ⅱ,Ⅳ,H-Y Scale,VAS as well as five sub-items of BPI including daily living,working,sleeping,walking ability and social communication [(13.7±5.3) vs.(12.3±6.3),(27.3±12.9)vs.(23.3±9.6),(2.3±2.2)vs.(1.7±1.3),(2.4± 1.0)vs.(2.1±0.9),(63.3±25.6)vs.(56.6±25.0),(5.3±2.7)vs.(4.6±2.7),(5.9±3.2)vs.(5.1±2.8),(6.3±2.5)vs.(5.6±2.6),(4.7±3.1)vs.(3.8±2.0),(3.2±2.1)vs.(2.6±2.5),t=1.976,2.539,2.287,2.381,2.050,2.021,1.997,2.165,2.420,2.134,respectively,all P＜0.05].No significant differences were found in SAS,SDS or other sub-items of BPI such as life pleasure and mood scores between the two groups (all P＞0.05).Compared with the young group,patients in the elderly group had a higher ratio of two or more pain types associated with PD[41.2％ (119/299)vs.23.4％ (18/77),x2=8.190,P＜0.05],but a lower ratio of pain-related treatment [29.76％ (86/299)vs.51.95％ (40/77),x2=13.260,P＜0.05].Conclusions Pain in elderlyPD patients is more severe,shows more diverse types,and significantly aftects the quality of life.Enhanced intervention is needed.

Objective To prospectively explore the impact of hypocalcemia on the prognostic value of intracerebral hemorrhage.Methods A total of 410 patients consecutively admitted within 12 hours after intracerebral hemorrhage onset were divided into 3 groups based on admission serum calcium:low serum calcium group,normal serum calcium group and high serum calcium group.Baseline characteristics of patients including age,gender,Glasgow coma score(GCS),hematoma volume,etc were collected and analyzed.A follow-up was performed after 6 months.Final outcome was assessed using Glasgow outcome scale(GOS)with a score＞3 regarded as favourable prognosis,a score≤3 as unfavourable prognosis.Results Patients with low serum calcium had lower GOS,bigger hematoma volume,higher rate of operation,higher re-bleeding rate,more unfavourable prognosis than did the other 2 serum calcium groups.Multivariable Logistic regression analysis showed that patients with low serum calcium had poorer prognosis than patients with normal serum calcium after adjusting for other potential confounders(Odds ratio:3.01,95％ confidence interval:1.06-6.12,P＜0.05).Conclusions Hypocalcemia is an independent risk factor for the prognosis of patients with intracerebral hemorrhage.

Semont maneuver was performed in 97 patients with benign paroxysmal positional vertigo involving the posterior semicircular canal.Among 97 patients the Semont maneuver was successful in 69 cases and failed in 28 cases.There were three nystagmus patterns during the third position of the Semont maneuver:orthotropic nystagmus (n=45), no nystagmus (n=42) and reversed nystagmus (n=10);and the effective rates in three groups were 93%, 64% and 0%, respectively (P<0.05).The duration of latency period and nystagmus status in Dix-Hallpike test has no effect on repositioning efficacy ( P>0.05) .

Objective To investigate the incidence trends,clinical features and prognosis of ischemic colitis (IC) in China,and improve the level of diagnosis and treatment of IC.Methods China National Knowledge Infrastructure and VIP Chinese Science and Technology Periodical Database were searched.The ending date of search was May 15,2014.Results A total of 324 references were found,which were published from 1982 to 2013.There were 9202 cases reported,3973 cases of males and 5229 cases of females,with the male/female ratio of 1 ∶ 1.32 and mean age of (63.6±7.8)years.The amount of references and cases reported began to rise after 2002.Estimated ratio of cumulative incidence was higher in the north than in other areas of China (x2 =1097.95,P=0.000).The most common IC accompanying diseases were hypertension,heart diseases,diabetes,hyperlipidemia and constipation.There were statistically significant differences in the accompanying diseases between different regions and different times.Drugs,enteroscopy,surgery and low blood volume might be the precipitating factors.Patients commonly complained of abdominal pain,diarrhea/ desiring to defecate and hematochezia.Computed tomography was feasible in detecting lesions.Colonoscopy was the main method for diagnosis.The lesions were most common located in the left half colon including sigmoid colon,descending colon and splenic flexure,with typically in a segmental manner.Pan-colon involvement or rectum involvement rarely occurred.Type of transient lesion was the predominant subtype,which was generally managed non-operatively with good prognosis.Different from the type of transient lesion,pathological changes in gangrenous type were located in the right half colon including hepatic flexure of transverse colon,ascending colon and ileocecal junction.Gangrenous type required prompt surgical intervention but the mortality was much higher.Conclusions Incidence of CI has been increasing year by year.Patients who complain of abdominal pain,diarrhea/desiring to defecate and hematochezia should be considered as IC,particularly in the elderly women and patients with cardiovascular disease.Diseased regions are mainly located in sigmoid colon,descending colon and splenic flexure.Transient colitis is the predominant subtype,which have good prognosis.