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ObjectiveTo investigate the effects of Yishen Daluo prescription (YSDL) on Ras homolog(Rho)/Rho-associated coiled-coil containing protein kinase(ROCK)signaling pathway in mice with experimental autoimmune encephalomyelitis (EAE) based on the silencing of β-arrestin1 gene. MethodSixty C57BL/6 female mice were randomly divided into a blank group, a model group, a virus group, a YSDL group, a virus + YSDL group, and a prednisone acetate group (hormone group). The EAE model was induced in mice except for those in the normal group. Adeno-associated virus(AAV)solution (150 μL, 1×1011 vg·mL-1) was injected into the tail vein of each mouse in the virus group and the virus + YSDL group on the 4th day of immunization. Drugs were administered on the 8th day of modeling. Specifically, normal saline was given to the mice in the normal group,the model group,and the virus group at 10 mL∙kg-1, prednisone acetate suspension to those in the hormone group at 3.9 g∙kg-1,and YSDL to those in other groups at 20 g∙kg-1 for 14 consecutive days. The mice were weighed and scored every day. The neurological function scores of mice in each group were recorded every day after immunization. Hematoxylin-eosin (HE) staining was used to determine the inflammatory response and lesion location in the brain tissues and spinal cord tissues of mice. The protein expression of β-arrestin1,Ras homolog gene family member A(RhoA), and Rho-associated coiled-coil forming protein kinase Ⅰ(ROCK Ⅰ) in spinal cord and brain tissues of EAE mice was determined by Western blot. ResultCompared with the model group, the virus group and the virus + YSDL group showed decreased neurological function scores (P<0.01),and the YSDL group also showed decreased neurological function scores(P<0.05). HE results showed that there was obvious inflammatory reaction in the central nervous system (CNS) of the model group, which was alleviated to varying degrees in other groups compared with the model group. Western blot results showed that compared with the blank group, the model group showed increased protein expression levels of β-arrestin1, RhoA, and ROCK Ⅰ in the spinal cord tissues (P<0.01). Compared with the model group, the virus group, the YSDL group, the virus + YSDL group, and the hormone group showed decreased protein expression levels of β-arrestin1, RhoA, and ROCKⅠ in the spinal cord tissues (P<0.01). Compared with the blank group, the model group showed increased protein expression levels of β-arrestin1, RhoA, and ROCK Ⅰ in the brain tissues (P<0.01). Compared with the model group, the virus group, the YSDL group, the virus + YSDL group, and the hormone group showed decreased protein expression level of β-arrestin1 in the brain tissues (P<0.01), and the virus group and the YSDL group showed decreased protein expression levels of RhoA, and ROCKⅠ in the brain tissues (P<0.05). Additionally, the virus + YSDL group and the hormone group showed decreased protein expression levels of RhoA and ROCKⅠ in the brain tissues (P<0.01). ConclusionYSDL can improve the clinical symptoms of EAE mice and improve the inflammatory response of CNS. The mechanism is presumably attributed to the fact that YSDL inhibits the expression of β-arrestin1 in CNS,thereby reducing the expression of Rho/ROCK signaling pathway. Furthermore, YSDL may have a synergistic effect with the inhibition of β-arrestin1 gene expression.
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Resumen La presente revisión sistemática tuvo como objetivo identificar y analizar las propiedades psicométricas de los instrumentos de alimentación emocional (AE) actualmente validados en México. La búsqueda de los test se realizó con base a los criterios PRISMA en tres bases de datos: Pubmed, Science Direct y Redalyc. Fueron identificados dos instrumentos: El Cuestionario sobre la Conducta Alimentaria Relacionada con las Emociones y el Estrés (EADES) y la Escala de Alimentación Emocional (EAE). El método de validez empleado en ambas escalas fue el análisis factorial exploratorio y de confiabilidad el alpha de Cronbach (α = .91 en el EADES y α =.94 en la EAE). Se concluye que el empleo de estas escalas podría estar en función de la definición conceptual y operacional de la AE que considera cada una, con el fin de retomar la que mejor se adecúe a los objetivos del estudio. Asimismo, se sugiere dar continuidad a la consolidación de las propiedades psicométricas de ambas escalas, a través de ampliar a otras poblaciones (clínicas, de diferentes etapas etarias y residentes de distintas zonas del país) así como emplear los actuales indicadores psicométricos sugeridos (entre ellos el AFC y cálculo del coeficiente omega).
Abstract The objective of this systematic review was to identify and analyze the psychometric properties of the emotional eating (EE) tests currently validated in Mexico. The search was performed according to PRISMA criteria in three databases: Pubmed, Science Direct and Redalyc. Two instruments were identified: The Eating and Appraisal Due to Emotions and Stress Questionnaire (EADES) and the Emotional Eating Scale (EAE). The validity method used in both scales was the exploratory factor analysis and reliability of Cronbach's alpha (α = .91 in the EADES and α = .94 in the EES). It is concluded that the use of these scales could be based on the conceptual and operational definition of the EE that each one considers, in order to retake the one that best suits the objectives of the study. In the same way, it's suggested to give continuity to the consolidation of the psychometric properties of both scales, through expanding to other populations (clinics, of different age groups and residents of different areas of the country) as well as using the currently suggested psychometric indicators (among them the CFA and the calculation omega's coefficient).
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@#【Objective】 To study the mechanisms and therapeutic effects of human olfactory mucosa-derived mesenchymal stem cells(OMSC)on experimental autoimmune encephalomyelitis(EAE)in mice.【Methods】Under local anesthesia by using nasal endoscopy,olfactory epithelia of healthy donors were obtained,digested and cultured up to the 5th passage. OMSC were identified,differentiated and stained. EAE models were induced in C57 female mice by myelin oligodendrocyte glycoprotein(MOG35- 55)and pertussis toxin(PT). Neurological function was documented daily. On day 16 after immunization(peak of incidence),the mice were divided randomly into two groups and treated with OMSC and PBS via tail vein injection respectively. On day 24 after immunization ,blood was collected from angular vein and levels of IL-10,IL-17,IFN-γ and IL-6 were determined by cytometric beads array(CBA). The size of the spinal cord lesion in mice was observed and measured by using HE and LFB staining. Peripheral blood lymphocytes(PBL)of healthy donors were obtained and then co-cultured with OMSC. The proportions of CD4+ T cells secreting IFN- γ(Th1 cells)in lymphocyte group and co-culture group were compared after 2 days of cultivation. Adding IDO or COX pathway inhibitor to co- culture group and cultivating for 2 days,we observed and compared the proportions of Th1 cells in lymphocyte group,co-culture group and inhibitor treatment group respectively.【Results】OMSC exhibited certain mesenchymal stem cell-like characteristics with respect to expression of stem cell surface markers and multilineage differentiation potentials. After induced by MOG35- 55 and PT,EAE models showed different levels of neurological damage. Compared with those in PBS treatment group,in OMSC treatment group,the severity of neural dysfunction in mice was significantly reduced(P =0.002),the level of IFN-γ in serum was lower(P = 0.032),but no significant differences in the levels of IL-10,IL-17 and IL-6 were found between two groups. HE and LFB staining revealed that the inflammatory infiltration and demyelinating areas in OMSC treatment group were less than those in PBS treatment group. The proportion of Th1 cells was lower in co-culture group than that in lymphocyte group(P = 0.001),higher in IDO inhibitor group than that in co-culture group(P = 0.01),but no significant difference was found between IDO inhibitor group and lymphocyte group or between COX inhibitor group and co-culture group.【Conclusions】OMSC may regulate the proportion of Th1 lymphocytes through IDO pathway so as to inhibit the demyelinating injuries of EAE in mice. This study provides a new idea for the clinical treatment of multiple sclerosis.
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Objective:To investigate the effect of miR-322-5p which targets Akt3 on Th17 differentiation in experimental autoimmune encephalomyelitis (EAE) interfered by interferon-β (IFN-β). Methods:The effect of IFN-β on EAE mice which were randomly divided into IFN-β group and PBS group was examine. The percents of Th17 in the two groups were compared by fluorescence activated cell sorting. The miRNA array was made to find different miRNAs between those two groups. MiR-322-5p was screened for further research. The target gene of miR-322-5p was predicted using softwares and the common predicted target gene Akt3 was got. The expression of Akt3 was detected after IFN-β intervention and miR-322-5p overexpression. The target relationship between Akt3 and miR-322-5p was verified by luciferase reporter assay. At last, the effect of Akt3 on Th17 differentiation was explored in vitro. Results:Compared to PBS group, the percent of Th17 was significantly downregulated, the expression of miR-322-5p was significantly upregulated and Akt3 was significantly downregulated in IFN-β group. The expression of Akt3 was obviously decreased after overexpressing miR-322-5p. Luciferase reporter assay showed that Akt3 was directly targeted by miR-322-5p. The percent of Th17 differentiation was greatly promoted by Akt3 in vitro. Conclusion:IFN-β significantly ameliorates the severity of EAE by affecting miR-322-5p which can inhibit Th17 differentiation by targeting Akt3.
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Flavonoids are widely found in medicinal plants, which have important medical properties. Flavonoids were proved to have many pharmacological activities, such as anti-oxidation, antitumor, antimutation, anti-inflammatory, antibacterial and anti-aging. The extraction of flavonoids is the crucial link in their clinical applications. In recent years, many emerging Chinese medicine extraction methods have also been widely used in the extraction of flavonoids. This paper reviews the current application of new methods for flavonoid extraction, in order to provide references for the extraction, development and utilization of flavonoids. These new extraction methods include supercritical fluid extraction (SFE), ultrasonic assisted extraction (UAE), microwave assisted extraction (MAE), pressurized liquid extraction (PLE), pulsed electric field (PEF) assisted extraction, enzyme assisted extraction (EAE), green solvent extraction, steam explosion assisted extraction, dynamic high pressure microfluidization (DHPM) assisted extraction, etc.
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Engajamento escolar refere-se ao nível de envolvimento do aluno em atividades escolares, envolvendo quatro aspectos: comportamental, cognitivo, afetivo e agente. Este estudo objetivou adaptar e obter evidências de validade da escala EAE-4DE para o contexto brasileiro. Participaram desta pesquisa 337 alunos do 5º e 6º anos do Ensino Fundamental de escolas públicas e privadas. Os dados foram submetidos à análise fatorial exploratória com o método de extração dos principais eixos fatoriais. O método das Análises Paralelas foi empregado, o que resultou em quatro fatores que explicaram 52,08% da variância. Os fatores foram teoricamente coerentes com as quatro dimensões do engajamento escolar previstas na escala original e apresentaram índices Alfa de Cronbach de aceitáveis (0,62 para o cognitivo) a bons (0,74, 0,78 e 0,81 para agente, comportamental e afetivo, respectivamente). Destaca-se a utilidade da escala para rastreio e monitoramento do engajamento na escola, mas recomenda-se cautela na interpretação do fator "engajamento cognitivo".
School engagement is understood as the student's level of involvement in school activities, consisting of four aspects: behavioral, cognitive, emotional, and agent. This study aimed at adapting and bringing validity evidence for the EAE-4DE scale in Brazil. Three hundred and thirty-seven students from the 5th and 6th grades of private and public elementary schools took part in this study. Factor Analysis by principal axis factoring extraction method and Parallel analysis method resulted in four factors. These four factors explained 52.08% of item variance and mirrored the four school engagement dimensions predicted by the original EAE-4DE scale. The cognitive engagement factor presented a fair Cronbach's alpha (.62), and the other factors presented good Cronbach's alphas: agentic (.74), behavioral (.78), and affective (.81). There was a highlight in the screening and pedagogical potential of the Brazilian version of the EAE-4DE scale. However, the scales' cognitive engagement factor calls for cautious interpretation.
Lo compromiso escolar se refiere al nivel de envolvimiento del alumno en actividades escolares las cuales envuelven cuatro aspectos: comportamental, cognitivo, afectivo y agente. Este estudio tiene como objetivo principal adaptar y obtener evidencias de validez de la escala EAE-4DE para el contexto brasileño. Participaron de esta investigación 337 alumnos del 5º y 6º año de Enseñanza Básica de escuelas públicas y privadas. Las informaciones fueron sometidas a análisis factorial exploratorio con el método de extracción de los principales ejes factoriales. El método de análisis paralelos fue empleado resultando en cuatro factores, explicaron 52,08% de varianza. Los facto- res fueron teóricamente coherentes con las cuatro dimensiones de la participación escolar previstas en la escala original y presentaron índices Alfa de Cronbach, de aceptables (0,62 para lo cognitivo) a buenos (0,74, 0,78 e 0,81 para agente, comportamental y afectivo, respectivamente). Se destaca la utilización de la escala EAE-4DE en la escuela, más se recomienda cautela en la interpretación del factor "Adaptación Cognitiva".
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Humanos , Masculino , Feminino , Criança , Adolescente , Psicologia , Escolaridade , Brasil , Análise Fatorial , Ensino Fundamental e MédioRESUMO
Objective:To investigate the therapeutic effects of astragalus polysaccharide (APS) on experimental autoimmune encephalomyelitis(EAE) and to explore the regulating effects on microglia activation that is associated with the pathogenesis of EAE and its possible mechanisms.Methods:Animal experiments:EAE model was induced by MOG35-55in C57BL/6 mice.APS was given by gavage.EAE was scored according to a 0-5 scale to observe the therapeutic effects of APS.Cell experiments:The effects of lipopolysac-charide (LPS) on cell viability of BV-2 microglial cell line were investigated by MTT assay and then the appropriate concentration of LPS to activate the BV-2 microglial cell line was selected.The microglia activation model was established.The changes in BV-2 microglial cell line morphology were observed with an inverted microscope.The cytokines of TNF-α and IFN-γ in the cell culture supernatant of BV-2 microglial cell line were detected by ELISA.The activated BV-2 microglial cells were treated with APS in different concentrations.The regulatory roles of the APS on the BV-2 microglial cell activation were observed.Western blot and Real-time PCR method were used to measure the protein and mRNA level of the PD-L1 on the cell surface of BV-2 microglial cells treated with APS.Results:APS could effectively ameliorate the symptoms in EAE mice and could suppress neuroinflammation of EAE significantly.The microglia activation model in vitro induced by LPS was successful.APS in certain concentration could inhibit the activation of microglia,increase the viabilily of the active microglia.Meanwhile,it could downregulated the level of the cytokines including IFN-γ and TNF-α and upregulated the expression of protein and mRNA of PD-L1 on activated microglia.Conclusion:APS can effectively inhibit the autoimmune reaction of EAE and effectively suppress the microglia activation induced by LPS,reduce the pro-duction of IFN-γ and TNF-α.APS plays a crucial role in reducing the inflammation induced by microglia activation.The potential mech-anisms might be related to the upregualtion of the PD-1/PD-L1 pathway.
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In recent years,it has been found that there are lymphoid follicle structures outside the lymphoid tissues and organs, which are called ectopic lymphoid tissues,also known as tertiary lymphoid tissues(TLTs).TLTs currently have been found in a number of diseases such as multiple sclerosis,diabetes,cancer,atherosclerosis and rheumatoid arthritis and the like.Here we focus on the molecular mechanism of TLTs and the relationship between TLTs and multiple sclerosis (MS)/experimental autoimmune encephalomyelitis (EAE) disease,and briefly describe the role of TLTs in the development of MS/EAE,the relevant mechanisms and research progress,make people have more understanding and insights on the formation of TLTs and the mechanism that it involved in disease,to provid theoretical knowledge for clinical treatment.
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Objective To explore the effects of vasoactive intestinal peptide (VIP)on the ratio of CD4+CD25+Treg/CD4+T cell and the expression of TGF-β1 in experimental autoimmune encephalomyelitis (EAE)rats. Methods We randomly divided 60 healthy female Wistar rats into normal control group,EAE control group,VIP low-dose group and VIP high-dose group.We used myelin basic protein (MBP)+ complete adjuvant (CFA)to establish EAE model. Since the day of model construction, the low- and high-dose VIP groups received intraperitoneal injection of 4 nmol/kg (0.2 mL)and 16 nmol/kg (0.8 mL)of VIP every other day,respectively;normal control group and EAE group received injection of saline of 0.8 mL for 10 days in a row.We recorded the peak of neurological dysfunction score (NDS)changes in the rats,observed the pathological changes and GFAP+astrocyte activation in the brain at the morbidity peak of rats with HE staining,and detected the ratio of CD4+CD25+Treg/CD4+T in the spleen with FACS and TGF-β1 cytokine level in brain tissue with ELISA.Results The peak nerve dysfunction score was decreased in each VIP dose group.In normal control group,there were decreased inflammatory cell infiltration and decreased number of active astrocytes in the brain tissue.The degree of infiltration of inflammatory cells and astrocyte activation in VIP control groups were significantly lower than those in EAE group.The CD4+CD25+Treg/CD4+T cell ratio of the spleen tissue in each dose VIP treated group rats was higher than that in EAE control group.The cytokine level of TGF-β1 in the brain tissue increased in each VIP dose group in the dose-dependent manner.Conclusion Through up-regulating the ratio of CD4+CD25+Treg/CD4+T cell in the spleen tissue,increasing TGF-β1 content in brain tissue,and inhibiting the infiltration of inflammatory cells and the astrocyte activation,VIP plays an important role in prevention and control of EAE.
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Multiple sclerosis (MS) is a T-lymphocyte-mediated autoimmune disease that is characterized by inflammation in the central nervous system (CNS). Although many disease-modifying therapies (DMTs) are presumed effective in patients with MS, studies on the efficacy and safety of DMTs for preventing MS relapse are limited. Therefore, we tested the immunosuppressive anti-inflammatory effects of oral-formulated tacrolimus (FK506) on MS in a mouse model of experimental autoimmune encephalomyelitis (EAE). The mice were randomly divided into 3 experimental groups: an untreated EAE group, a low-dose tacrolimus-treated EAE group, and a high-dose tacrolimus-treated EAE group. After autoimmunization of the EAE mice with myelin oligodendrocyte glycoprotein, symptom severity scores, immunohistochemistry of the myelination of the spinal cord, and western blotting were used to evaluate the EAE mice. After the autoimmunization, the symptom scores of each EAE group significantly differed at times. The group treated with the larger tacrolimus dose had the lowest symptom scores. The tacrolimus-treated EAE groups exhibited less demyelination and inflammation and weak immunoreactivity for all of the immunization biomarkers. Our results revealed that oral-formulated tacrolimus inhibited the autoimmunization in MS pathogenesis by inactivating inflammatory cells.
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Animais , Humanos , Camundongos , Doenças Autoimunes , Biomarcadores , Western Blotting , Sistema Nervoso Central , Doenças Desmielinizantes , Encefalomielite Autoimune Experimental , Imunização , Imuno-Histoquímica , Inflamação , Esclerose Múltipla , Bainha de Mielina , Glicoproteína Mielina-Oligodendrócito , Neuromielite Óptica , Recidiva , Medula Espinal , TacrolimoRESUMO
Objective:To investigate the role of B cell adoptor protein with ankyrin repeats ( BANK ) in experimental autoimmune encephalomyelitis ( EAE) .Methods: C57BL/6 mice and BANK-deficient ( BANK-/-) mice were immunized with MOG peptide in CFA,and then observed the clinical symptoms and pathological severity .Results: The percentages of CD4+T cells,CD8+T cells and regulatory T cells in brain and spleen were analyzed by flow cytometry .BANK-/-mice showed significantly higher score at the peak and the plateau phase compared with wild-type mice(P<0.05).HE staining showed more widespread areas of inflammation and demyelination in BANK-/-mice when compared to wild-type mice on day 16.In addition,the frequency of CNS-infiltrating CD8+T cells was markedly higher in BANK-/-mice than in wild-type mice.In addition,the percentage of CD8+T cells from spleen in BANK-/-mice was also increased compared with wild-type mice (P<0.05).By contrast,the percentage of regulatory T cells and the ratio of CD 4/CD8 T cells from spleen in BANK-/-mice were significantly lower than in wild-type mice(P<0.05).Conclusion:Thus,BANK expression in B cells can inhibit the development of EAE .
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We investigated the carrying situation of Escherichia albertii from healthy people engaged in breeding and slaughtering poultry for a long time.We collected stool samples from people engaged in breeding and slaughtering poultry and other healthy people.After enriched with EC broth,eae-positive enrichment culture was directly streaked on MacConkey,and eaepositive lactose non-fermenting isolate was retained for further investigation.The 16S rDNA sequencing and multilocus sequence typing (MLST) were applied in the identification of E.albertii from suspected strains.Intimin subtypes and cdtB types of E.albertii strains were detected.Pulsed-field gel electrophoresis (PFGE) was used to detected genetic polymorphism of strains from this study and animal source ones.Results showed that two isolates were identified as E.albertii from 189 stools of people exposed to slaughtering chickens and ducks and one from 58 stools in control groups.No isolate was identified as E.albertii from 138 stools samples of people exposed to breeding poultry.Intimin subtypes of three isolates from stool samples were subtyped as sigma,iota 2,nu,and cdtB types were closely related to types Ⅱ/Ⅲ/Ⅴ.PFGE patterns of the three strains was distinguishable (<80% similarity),and appeared in different cluster with chickens,ducks and other sources of E.albertii strains.The rate of carrying E.albertii to a certain extent exist in healthy people engaged in slaughtering chickens and ducks,and the relationship between these strains and strains from poultry should be further investigated.
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BACKGROUND: CD4+ T cells play an important role in the initiation of an immune response by providing help to other cells. Among the helper T subsets, interferon-γ (IFN-γ)-secreting T helper 1 (Th1) and IL-17-secreting T helper 17 (Th17) cells are indispensable for clearance of intracellular as well as extracellular pathogens. However, Th1 and Th17 cells are also associated with pathogenesis and contribute to the progression of multiple inflammatory conditions and autoimmune diseases. RESULTS: In the current study, we found that BJ-1108, a 6-aminopyridin-3-ol analogue, significantly inhibited Th1 and Th17 differentiation in vitro in a concentration-dependent manner, with no effect on proliferation or apoptosis of activated T cells. Moreover, BJ-1108 inhibited differentiation of Th1 and Th17 cells in ovalbumin (OVA)-specific OT II mice. A complete Freund's adjuvant (CFA)/OVA-induced inflammatory model revealed that BJ-1108 can reduce generation of proinflammatory Th1 and Th17 cells. Furthermore, in vivo studies showed that BJ-1108 delayed onset of disease and suppressed experimental autoimmune encephalomyelitis (EAE) disease progression by inhibiting differentiation of Th1 and Th17 cells. CONCLUSIONS: BJ-1108 treatment ameliorates inflammation and EAE by inhibiting Th1 and Th17 cells differentiation. Our findings suggest that BJ-1108 is a promising novel therapeutic agent for the treatment of inflammation and autoimmune disease.
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Animais , Feminino , Camundongos , Diferenciação Celular/efeitos dos fármacos , Células Th1/efeitos dos fármacos , Encefalomielite Autoimune Experimental/tratamento farmacológico , Células Th17/efeitos dos fármacos , Aminopiridinas/farmacologia , Compostos de Anilina/farmacologia , Baço/imunologia , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Reprodutibilidade dos Testes , Apoptose/efeitos dos fármacos , Apoptose/imunologia , Células Th1/imunologia , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/imunologia , Células Th17/imunologia , Citometria de Fluxo , Aminopiridinas/imunologia , Compostos de Anilina/imunologia , Linfonodos/imunologia , Camundongos Endogâmicos C57BLRESUMO
Background: Animal model of multiple sclerosis is a demyelinating and inflammatory disorder of central nervous system and eye. Histological evaluation in eyes in experimental autoimmune encephalomyelitis (EAE) models demonstrated evidence of retinal atrophy and inflammation in late stage of disease. Deciphering the relationships between the retinal atrophy and proliferation on retinal layers may help us in understanding the factors that drive atrophy and proliferation in multiple sclerosis. Aims: The aim of the present study was to determine alterations in thickness of retina and its sub-layers in MS induced mice model in comparison with control group. Study Design: Experimental study. Methodology: EAE was induced in female C57BL/6 mice using Hooke kits. Animals were scored for clinical signs of the disease according to 10-point EAE scoring system. At 35th day after immunization, mice (n=15/group) were deeply anesthetized and eyes were removed. Morphometric study of proliferation in retinal sub-layers were assessed by Hematoxilen and Eosin staining and expression of Ki67. The proliferating marker was performed by Ki67analyzing kit. All measurement obtained by Motic image analyzer software 2 and analyzed spss 18, respectively. Results: Here we reported that retinal thickness in MS group including total retinal layer, especially photoreceptor layer, ganglion cell layer and neural layer reduced in comparison to control group (p< 0.001). In Ms Group proliferation rate is also decreased in comparison to control group (0.05). Conclusion: Our results show that ki67 expression, as an indication of proliferation, decreased in retinal layers in MS group. Furthermore, our data revealed that retinal thickness also decreased in MS group.
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Objective:To investigate the change of thymus in different period of EAE mice. Methods: The thymic index and thymocyte number at 0,10,15,20 days after EAE induced were recorded. The percent of thymic apoptosis and CD4+CD44+T cells in spleen was acquired by Flow Cytometry. The content of p53 and Bcl-2 gene was detected by PCR and electrophoretic technique. Results:The thymic index and thymocyte number correspondingly reduced at 0,10,15 days after EAE induced. Apoptotic rate of thymocyte increased at 10,15 days after EAE induced,highest at 10 days. The percent of CD4+CD44+T cells also increased at 10,15 days after EAE induced, highest at 10 days. The content of p53 increased, while Bcl-2 reduced at 10 days after EAE induced. Conclusion:The atrophy of thymus is most serious at 15 days after induction;while the apoptotic percent is highest at 10 days after induction,which have a relationship with regulation of p53 and Bcl-2. The change of thymus in EAE mice closely related with EAE attack,and the recovery of thymus precede EAE.
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Objective: To evaluate the mechanism of T follicular helper cells ( Tfh ) in experimental autoimmune encephalomyelitis (EAE) via in vivo experiments. Methods:C57BL/6 mice were randomly divided into four groups,CFA group,EAE group,anti-ICOSL group and control group. Lymphocytes of different time points isolated from draining lymph nodes and spleen were stained for T follicular helper cells surface marker and T cells activation surface marker and analyzed by FACS. Observed parameters include inflammatory infiltration,demyelination in spinal cord and germinal center in spleen. ELISA was used to measure the level of antigen specific antibodies. Results: Mice in anti-ICOSL treated group developed mild disease was with lower clinical scores when compared with the EAE group. HE staining results turned out with alleviated inflammation and Luxol Fast Blue staining( LFB) showed no demyelization in anti-ICOSL treated mice compared with non-treated EAE models. Flow cytometry results revealed that percentages of T follicular helper cells decreased though the whole activated degree T cells was not influenced in anti-ICOSL treated group. Fewer ger minal center was found in both anti-ICOSL group and CFA group with reduced secretion of MOG-specific Ab. Conclusion:T follicular helper cells supported the development of cognate B cells,promoted the formation of germinal center,facilitate pathogenic MOG-specific Ab secretion,thus enhance EAE.
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Objective To explore the appropriate dosage of drugs inducing experimental allergic en cephalomyelitis (EAE) in mice,and evaluate the modified model mice.Methods Different doses of myelin oligodendrocyte glycoprotein (MOG35-55:200 μg,100 μg,50 μg,25 μg),together with different doses of inactivation of mycobacterium tuberculosis (H37RA:800 μg,250 μg,100 μg) and pertussis toxin (500 ng,200 ng),were used to induce the EAE model.After immunized,the clinical disease severity of EAE mice was measured by the standard EAE grading clinical score daily.The open field test was used to detect the locomotion of mice.The Western blot and immunofluorescence were used to detect the level of myelin basic proteins (MBP) in different brain regions of mice.Results Compared with the EAE mice induced with high-dose drugs,the mice with low-dose drugs (25 μg MOG35-55,100 μg H37RA,200 ng pertussis toxin) had low neu rological scores.And they displayed normal locomotion compared with the control mice (day 16:group EAE (8.885±0.772) cm/s vs control group (8.933±0.567) cm/s,P>0.05;day 31:group EAE (11.130±0.630) cm/s vs control group (10.670±0.959) cm/s,P>0.05;day 55:group EAE (7.686±0.428) cm/s vs control group (8.313±0.918) cm/s,P>0.05).Moreover,there was a significant decrease of MBP in the parahippocampal cortex (PHC) and fimbria-fornix of EAE mice induced with low-dose of drugs (PHC:group EAE (0.369±0.096) vs control group (1.000±0.163),P<0.05;fimbria-fomix:group EAE (0.494±0.071) vs control group (1.000±0.143),P<0.05).Conclusion The EAE mice induced with low-dose drugs(25 μg MOG35-55,100 μg H37RA,200 ng pertussis toxin) have low neurological scores,normal locomotion,and myelin impairment in the central neuronal system.And it can be used in the cognitive behavioral research of demyelination disease,such as multiple sclerosis.
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Experimental autoimmune encephalomyelitis (EAE) is an autoimmune disease model mediated by specific allergenic CD4GT cell and accompanied by mononuclear cells invasion in central nervous system (CNS) and demyelination. Similar to the histopathology and immune characters of multiple sclerosi (MS), EAE has been used as a classical animal model for the study of MS. In recent years, as an immune regulator for the treatment of relapsing-remitting MS, study of estrogen has entered the preclinical phase H trials. Larger numbers of studies have verified that high concentration of estrogen could improve EAE. However, the specific mechanisms are still in controversy. For these considerations, this review summarizes the protective mechanisms of estrogen in EAE.
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Objective To analyze the subtypes of eae genes in various non-O157 Shiga toxin-pro-ducing Escherichia coli ( STEC) strains isolated in China.Methods The complete nucleotide sequences of 10 eae genes were amplified by PCR and sequenced.The BLASTn software was used to analyze the se-quences for eae gene subtyping.A phylogenetic tree was constructed based on the10 ea e gene sequences to-gether with the gene sequences of 30 different subtypes in GenBank and those of STEC strains of 7 prevalent serotypes (O157 ∶H7, O26 ∶H11, O103 ∶H2, O111 ∶H8, O145 ∶H28, O45 ∶H2 and O121 ∶H19) using MEGA 5.0.Multilocus sequence typing (MLST) was performed on the 10 STEC strains with reference to the Escherichia coli ( E.coli) MLST website ( http://mlst.warwick.ac.uk/mlst/dbs/Ecoli) for the typing of multiple loci.A minimum spanning tree ( MST) was constructed using the BioNumerics software to inves-tigate the phylogenetic relationships between the 10 eae gene-positive STEC strains in this study and hemolyt-ic uremic syndrome-associated enterohemorrhagic E.coli ( HUSEC) strains as well as all human STEC strains of O157, O26, O45, O103, O111, O121 and O145 serotypes submitted to the E.coli MLST website data-base.Results The complete nucleotide sequences of eae genes in 10 non-O157 STEC strains were 2.8 kb in length and belonged to 3 known subtypes.The predominant subtype wasβ1, accounting for 60%of the 10 STEC strains (6/10), followed byθandγ1 subtypes with two strains in each type.The eae gene sequences in certain strains were identical to those of the prevalent serotypes.Seven sequence types ( STs) were identi-fied from the 10 STEC strains carrying eae gene.Conclusion The eae genes harbored by the non-O157 STEC strains isolated from different specimens in China were diverse and had close phylogenetic relationships with the highly pathogenic and prevalent STEC strains.This study implied that the STEC strains harboring eae gene had high pathogenic potential.
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Objective: To investigate whether Irgm 1 impact CD4+T cell subsets in the experimental autoimmune encephalomyelitis.Methods: The Irgm1 heterozygous mice were backcrossed with C 57BL/6 Wt.mice for 10 generations to produce C57BL/6 Irgm1+/-mouse.C57BL/6 Irgm1+/-mice were intercrossed to obtain three genotypes:Irgm1-/-, Irgm1+/-and Irgm1+/+.To establish model of EAE ,C57BL/6 Wt.mice and Irgm1 knock out mice were immunized with myelin oligodendrocyte glycolprotein 33-55 ( MOG33-55 ) and the clinical symptoms were observed.The proliferation of lymphocytes to MOG antigen was detected with Methyl Thiazolyl Tetrazolium ( MTT).The infiltration of inflammatory cells in the spinal cords was observed through HE staining.The CD4+T cell subsets from lymph nodes ,spleens and CNS were detected by flow cytometry.Results:EAE model was induced successfully.The proliferation of T cells in lymph nodes in Irgm 1-/-mice was lower than Wt.mice.Quantitative analysis of flow cytometry indicates that , compared with Wt.mice,the level of Th1 subset was higher,Th17 was lower relatively in lymph nodes and CNS of Irgm1 knock out mice.Conclusion:Irgm1 knockout mice can be partially protected EAE spinal cord function and clinical symptoms .Irgm1 may play a key role at early stage of EAE ,which may use as an important molecular target for treatment of EAE.