96% 7-10 days after culture. Immunohistochemistry of Ⅷ factor polyclone antibody showed positive in cytomembrane and cytoplasm, but negative in cell nucleus. These were cells with blood-brain barrier. CONCLUSION: Ideal endothelial cells of blood-brain barrier can be cultured by the improved cultural method of double filtering.

Objective To investigate the phenotypes and genetics of an early-onset familial Alzheimer′s disease ( EO-FAD ) family.Methods The clinical manifestations , brain MRI results and neuropathological findings of the proband and pedigree members of the EO -FAD family were evaluated. Autopsy was performed in the proband . Results Fifteen members of this family had a presenilin 1 (PSEN1) p.G378E mutation and nine of them had clinical manifestations or the MRI changes of EO -FAD. Neuropathological findings from autopsy of the proband disclosed moderate cortical atrophy throughout the brain, especially in frontal lobe and temporal lobe .Neuronal loss with gliosis was observed in the cortices of the frontal, temporal and occipital lobes , as well as in parahippocampal gyrus .Numerous senile plaques and neurofibrillary tangles were present in the cerebral cortex .The proband′s younger sister showed similar clinical presentations and MRI changes , and other members of this family demonstrated progressive memory loss.Conclusion A p.G378E mutation in the PSENl gene was identified in a Chinese EO-FAD pedigree.

Objective To determine the expression profile of serum microRNAs(miRNAs) in early-onset familial Alzheimer's disease (EO-FAD) patients. methods miRNA microarrays were performed to detect the expression profile of serum miRNAs in 2 cases of EO-FAD patients,2 cases of EO-FAD carriers and 2 cases of normal controls.Preliminary bioinformatic analysis was conducted. Result sIt was found that 21 miRNAs were up-regulated and 22 miRNAs were down-regulated in serum of EO-FAD patients,the differences were statistically significant(P<0.05).miR-5704(P=0.0002),miR-4639-3p(P=0.0195),miR-107(P=0.0204) were markedly up-regulated,miR-5572(P=0.0008),miR-204-3p(P=0.0014),miR-542-5p(P=0.0106) and miR-155-5p(P=0.0240) were markedly down-regulated.Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis suggested that the dysregulated miRNAs may be involved in the mechanism of EO-FAD by affecting neurotrophin signaling pathway.Conclusion miR-5704,miR-4639-3p,miR-107,miR-5572,miR-204-3p,miR-542-5p and miR-155-5p may be used as potential biomarkers of EO-FAD,and involved in the mechanism of EO-FAD by affecting neurotrophin signaling pathway.

N6-methyladenosine (m 6A), one of the most common types of eukaryotic mRNA methylation modification, is widely distributed in the nervous system and participates in important regulatory mechanisms for controlling gene expression. With the participation of related methylation-modifying enzymes and proteins, m 6A plays an important role in the growth and development of the nervous system, related functions, and the occurrence and development of diseases in the nervous system by affecting the "life cycle" of mRNA. This article briefly reviews the research progress on the role of mRNA m 6A methylation modification in neurological diseases and nervous system growth and development.

Both multiple sclerosis (MS) and neuromyelitis optica spectrum disorders (NMOSDs)are the most common demyelinating diseases of the Central Nervous System.Hormone,intravenous immunoglobulin and plasma exchange seem to be an effective therapy during MS and NMOSD acute phase;however,immunomodulators and immunosuppressive agents are conventional treatments for MS in the remission period,in addition monoclonal antibody,intravenous immunoglobulin G,estrogen,statins,vaccination and traditional Chinese medicine have also been gradually applied to clinical practice.Combined maintenance therapy with low-dose hormones and immunosuppressive agents will benefit NMOSD patients.Because of the different pathogenesis,MS and NMOSD have obvious differences in treatment.This review mainly focus on progress in clinical treatment of multiple sclerosis and neuromyelitis optica spectrum disorders.

Objective To investigate the expression of ferroptosis-related proteins in the hippocampus in AD mice. Methods AD mice were created by knocking in PSEN1 p.G378E gene in C57BL/6 mice. The expression of ferroptosis-related proteins, glutathione peroxidase 4 (GPX4), cystine-glutamate exchanger (SLC7A11), long-chain fattyacyl-CoA synthetase 4 (ACSL4) and phosphatidylethanolamine-binding protein1 (PEBP1) in the hippocampus were compared among PSEN1 p.G378E-/-、 PSEN1 p.G378E-/+and WT mice. Results The expression levels of GPX4, PEBP1, SLC7A11 and ACSL4 proteins in the hippocampus were significantly increased in PSEN1 p.G378E-/-mice than in either PSEN1 p.G378E-/+or WT mice (P<0.05). Meanwhile, the expression levels of GPX4 and PSEN1 proteins were higher in PSEN1 p.G378E-/+mice than in WT mice (P<0.05). However, there were no significant differences in expression levels of SLC7A11 and ACSL4 proteins between PSEN1 p.G378E-/+mice and WT mice (P>0.05). Conclusion Expression levels of ferroptosis-related proteins are significantly increased in the hippocampus of PSEN1 p.G378E knock-in mice.

Objective To study the association between the single nucleotide polymorphism (SNP) of SEPT14 (rs77231105,rs10241628,rs11981883,rs73701167) and sporadic PD in Chinese Han populationin of Southwest Shandong.Methods One hundred and eighty PD patients from Southwest Shandong served as PD patient group and 200 healthy subjects from Southwest Shandong served as control group in this study.The distribution frequencies of alleles and genotypes in SNP of rs77231105,rs10241628,rs11981883 and rs73701167 were compared by PCR and sequencing respectively.Results No significant difference was found in the distribution frequencies of alleles and genotypes in rs77231105,rs10241628,rs11981883 between the two groups (P＞0.05).The distribution frequency of rs73701167 was significantly higher in PD patient group than in control group (31.1％ vs 20.5％,OR=1.75,95％CI=1.261-2.428,P=0.001).Conclusion The SNP of SEPT14 rs77231105,rs10241628 and rs1198188 are not associated with PD,the SNP of rs73701167 is associated with PD in Chinese Han population of Southwest Shandong.The allele C is a risk factor for PD.

At present, many drugs were developed based on the main pathological feature of Alzheimer′s disease (AD): "β-amyloid cascade hypothesis and abnormal tau protein aggregation" as targets, but the efficacy is unsatisfactory. With the progress on the study of pathological mechanism of AD, the role of microglia and their related expression genes, such as TREM2, CD 33, ABCA7 gene and their related signal transduction pathways in the pathological mechanism of AD has been paid more and more attention. The study on AD biomarkers and therapeutic targets based on microglia and their related expression genes has also increased significantly. This review will mainly focus on the pathophysiology of microglia, the mechanism of microglia in AD, the biomarkers related to microglia and the drug treatment of AD.

Objective:To compare the expression of myeloid cell trigger receptor expressed on myoid cell 2 (TREM2) in different brain regions of tyrosine kinase binding protein(TYROBP) knockout mice and wild-type mice at different months of age, and to explore the relationship between TREM2, TYROBP and early onset Alzheimer's disease(EOAD).Methods:Healthy TYROBP gene knockout mice were divided into three groups according to the results of gene sequencing: the homozygous (TYROBP -/-) group, the heterozygous (TYROBP -/+ ) group, and the wild type (WT) group.Western blot and RT-qPCR were used to detect the expression of TREM2 in prefrontal cortex and hippocampus of 2, 4 and 6 month old mice in the three groups and with 10 in each group at each time point. Results:(1) In the prefrontal cortex: Western blot and RT-qPCR results showed that compared with WT mice (2-month-old: (0.993±0.048), (1.654±0.033); 4-month-old: (0.503±0.019), (2.169±0.023); 6-month-old: (0.600±0.036), (1.468±0.057)), the levels of TREM2 protein and mRNA in 2-month-old TYROBP -/+ group ((0.746±0.062), (1.137±0.067)) and TYROBP -/- group ((0.661±0.028), (0.644±0.012)) were decreased.While in 4-month-old and 6-month-old TYROBP -/+ group (4-month-old: (1.140±0.006), (5.483±0.088); 6-month-old: (0.827±0.043), (3.020±0.082)) and TYROBP -/- group (4-month-old: (1.071±0.010), (3.012±0.150); 6-month-old: (0.627±0.026), (1.633±0.027)) were increased, especially in 4-month-old mice and the differences were statistically significant ( F=12.946, 134.445; 725.318, 289.202; 12.172, 202.791; all P<0.05). (2) In the hippocampus: Western blot results showed that compared with WT mice (2-month-old: (1.268±0.036); 4-month-old: (0.813±0.010); 6-month-old: (0.312±0.021)), the level of TREM2 protein in 2-month-old TYROBP -/+ group ((0.804±0.034)) and TYROBP -/- group ((0.534±0.020)) were decreased.While in 4-month-old and 6-month-old TYROBP -/+ group ((0.932±0.011); (0.769±0.031)) and TYROBP -/- group ((0.910±0.014); (0.609±0.018)) were increased, especially in 4-month-old mice and the differences were statistically significant ( F=142.807; 27.884; 94.067; all P<0.05). Conclusion:The expression level of TREM2 decreases in 2-month-old TYROBP gene knockout mice while increases in 4-month-old and 6-month-old TYROBP gene knockout mice.It is presumed that TREM2/TYROBP signal pathway participates in the pathological process of EOAD and plays different roles in different pathological stages of EOAD.

Objective:To explore the effects of behavior intervention on patients with ischemic cerebrovascular disease recurrence, carotid artery stenosis recurrence, and quality of life after carotid artery stenting implantation.Methods:Sixty patients with carotid stenosis who underwent stenting surgery between January 2017 and July 2018 in Affiliated Hospital of Jining Medical University were recruited.The subjects were randomly divided into behavioral intervention group and control group.The control group was routinely followed up after carotid artery stenting implantation.The behavioral intervention group added home visit, education, guidance of the control and detection of risk factors for cerebrovascular disease of stent implantation.Results:After 12 months of intervention, the incidence of ischemic cerebrovascular disease and the recurrence rate of carotid restenosis in the behavioral intervention group(6.7%, 3.3%) were significantly lower than those in the control group(30.0%, 20.0%) (both P<0.05). Six months after the intervention and 12 months after the intervention, the self-management ability score (intervention group: six months after the intervention (171.20±18.43), 12 months after the intervention (179.90±14.34); control group: six months after the intervention (160.77±13.43); 12 months after the intervention (164.27±14.85)) and quality of SS-QOL score (intervention group: 6 months after intervention (188.47±16.25), 12 months after intervention (203.17±13.84); control group: 6 months after intervention (170.67±15.82); 12 months after intervention (183.80±18.19)of the intervention group were higher than that of the control group, the difference is statistically significant (all P<0.05). Conclusion:Behavioral intervention after carotid artery stenting implantation can effectively reduce the incidence of stroke and the recurrence rate of carotid stenosis and improve the prognosis of patients.The mechanism may be related with that the behavioral intervention can improve the self-management ability and the quality of life of patients with carotid stenosis.