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【Objective】 To investigate the association between the long-stranded non-coding ribonucleic acid (lncRNA) MRAK088388 and allergic asthma in children. 【Methods】 A total of 15 healthy children and 15 children with asthma were monitored for disease progression over a 2-year period. Blood samples were collected from patients during the chronic phase of the disease for lncRNA/mRNA expression microarray analysis. Competing endogenous RNA networks (MRAK088388/miR-30a/ATG5) were identified by bioinformatics analysis. In vitro cultured bronchial epithelial (16HBE) cells were used to quantify gene and associated protein expression levels by real-time fluorescent quantitative polymerase chain reaction (qPCR) and protein blotting, respectively. Cell Counting Kit-8 and transwell assays were used to assess the proliferation and migration of 16HBE cells and verify the effects of MRAK088388, miR-30a and ATG5 on asthma. 【Results】 Six lncRNA-miRNA-mRNAs were identified by correlation analysis. By qRT-PCR analysis, MRAK088388/miR-30a/ATG5 was selected to construct the ceRNA network in this study. mRAK088388 and ATG5 expressions were high in the peripheral blood of children with asthma, while the expression of miR-30a was low (P<0.05). The expression level of E-cadherin was significantly higher in 16HBE cells after si-MRAK088388+TGF-β1 group, while the expression levels of Vimentin and α-SMA were significantly lower (P<0.05), indicating that knockdown of MRAK088388 inhibited the epithelial mesenchymal transition in 16HBE cells. Compared with si-NC+ TGF-β1 group, the cell morphology of si-MRAK088388+TGF-β1 group was similar to that of the control group, indicating that MRAK088388 knockdown attenuated TGF-β1-induced cell morphological changes; in addition, MRAK088388 knockdown inhibited TGF-β1-induced proliferation and migration of 16HBE cells. MRAK088388 was confirmed by qPCR and protein blotting to promote the progression of childhood asthma by targeting the miR-30a/ATG5 axis. 【Conclusion】 Childhood asthma is associated with the MRAK088388/miR-30a/ATG5 axis, and MRAK088388 is involved in the process of childhood allergic asthma by negatively regulating miR-30a expression and regulating elevated ATG5 expression levels to affect bronchial epithelial cell mesenchymal transition, proliferation, and migration.
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Objective To investigate the long non-coding RNA(lncRNA) MRAK08838 regulates macrophage function to influence the development of asthmatic airway inflammation. Methods MRAK088388 gene knockout (MRAK088388-/-) mouse model was prepared and allergic asthma was induced by dust mite protein Dermatophagoides farinae 1 (Der f1). The mice were sacrificed after 28 days of modeling, and serum was collected to measure IgE and IgG. The FinePointe RC system was used to measure airway hyperresponsiveness and evaluate lung function in mice. Lung tissue was taken for HE staining, and periodic acid-Schiff (PAS) staining was used to evaluate inflammatory infiltration and mucus secretion in mouse lungs. Fluorescence quantitative PCR was used to detect the expression level of lncRNA MRAK08838 in bronchoalveolar lavage fluid (BALF) cells and lung tissue of asthmatic mice. ELISA was used to detect the levels of inflammatory cytokines IFN-γ, IL-4, IL-5, IL-13, IL-10 and IL-17A. Flow cytometry was used to evaluate the phenotype of macrophages in BALF and lung tissue, as well as the proportion of neutrophils, eosinophils, and alveolar macrophages. The changes of the above indicators were detected in mice by adoptive transfer of bone marrow-derived macrophages (BMDM). Results Under the challengle of Der f1, MRAK088388-/- mice showed reduced allergic airway inflammation, including reduced eosinophils in BALF and reduced production of IgE and IgG1. In addition, Der f1-treated MRAK088388-/- mice had fewer M2 macrophages than wild-type asthmatic mice. Wild-type mouse BMDM (M0) and Der f1-treated MRAK088388-/- mice also showed mild inflammatory response. Conclusion Knockout of MRAK088388 alleviates airway inflammation in asthmatic mice by inhibiting M2 polarization of airway macrophages.
Assuntos
Animais , Camundongos , Camundongos Knockout , RNA Longo não Codificante/genética , Asma/genética , Macrófagos , Imunoglobulina ERESUMO
Objective To investigate the clinical value of serum lipoprotein ( a ) concentration in evaluation of plaques characteristics for patients with coronary atherosclerotic heart diseases ( CAHD ) . Methods Using case-control method, Patients with suspicious CAHD, received coronary computed tomography angiography in the Shanghai East Hospital during October 2013 to June 2015 were enrolled.According to the results of coronary artery CTA, the patients were divided into two groups : the CAHD group (352 cases) and control group(438 cases) , the particle concentrations and mass concentration of lipoprotein(a), triglyceride, total cholesterol, HDL-C, LDL-C, glucose, HBA1c and hs-CRP and other tests were measured, the patients of CAHD group were divided into three subgroups by characteristics of coronary artery plaques including soft plaque (176 cases), calcified plaque (90 cases) and mixed plaque (86 cases), analysis were made with all these data.Using T test or variance analysis to compare the means between or among groups, the risk for CAHD was analyzed by logistic regression, the relationship between LP (a) -P and LP( a) -M were explored by linearly egression analysis, Conformance test were analyzed using kappa test.Results Compared with control group, the mean results of the CAHD group are significantly higher than that of control group, including LP (a) -P 18.5(8.3 -43.0))nmol/L vs.13.6 (7.6-32.4)nmol/L( t =-2.110), LP(a)-M 183(71 -361)mg/L vs.126(67 -293)mg/L(t =-2.063), age (62 ±9)years vs.(52 ±9)years(t=-7.691), hs-CRP 0.86(0.44-1.97) )mg/L vs.0.70(0.38-1.64)mg/L(t=-2.236), glucose (6.1 ±2.29 )mmol/L vs.(5.36 ±1.32 )mmol/L(t=-4.914), BA1c (6.13 ±0.98) % vs.(5.81 ±0.58) %(t=-4.842), APO(B) (1.09 ±0.33) g/L vs.(1.03 ±0.29) g/L( t=-2.407), all of the P values <0.05;The relative risk(RR)of age, glucose, LP( a)-P and LP ( a)-M are 1.067, 2.377, 1.384 and 1.342 respectively; Among the three types of plaques groups,the mean differences of age, TC, HDL-C, LDL-C and LP ( a)-P are statistically significant ( F=6.276,3.060,3.127,4.723,2.878;all of the P<0.05);The median of LP ( a)-P in the soft plaque group 20.3(8.3-48.2)nmol/L is higher than that of the mixed plaque group 15.7(7.3-26.0)nmol/L(P<0.05 ) and calcified plaque group 15.6 ( 8.1 -23.1 ) nmol/L ( P <0.05 ).The linearly regression equation of LP ( a) -M and LP( a)-P is Y=6.646X, r=0.939; Consistency test indicate the two methods are not consistent when used for grouping ( Kappa value is 0.557 ).Conclusions Serum concentration of lipoprotein(a) is an independent risk factor of CAHD, and the particle concentration of LP(a) is closely related to the characteristics of the plaques, especially to the soft plaque.
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Objective To explore the serum level of Glucagon like peptide-1 in late-onset Alzheimer′s patients and its clinical significance.Methods Case control study.Collecting cerebral vascular disease fifty-five cases, diagnosed with late-onset Alzheimer′s disease sixty-one cases, type 2 diabetes mellitus fifty-one cases , type 2 diabetic patients combined with late-onset Alzheimer′s disease thirty-seven patients from the Shanghai East Hospital and partly Pudong area elderdly hospital during October 2013 to March 2014, and forty healthy persons as normal control from physical examination center of Shanghai East Hospital during September 2013 to February 2014.Measuring the concentrations of GLP-1,β-amyloid, Tau protein and other routinely used clinical tests in the serum of patients from the normal controls , cerebrovascular disease , late-onset Alzheimer′s disease, type 2 diabetes and type 2 diabetes mellitus combined with late-onset Alzheimer′s disease by ELISA method developed in our laboratory.The blood samples were also collected at three fixed time including fasting time ,1 hour after taking glucose , 2 hour after taking glucose, the concentrations of GLP-1 were determined in the LOAD group , T2DM group and the T2DM combined with LOAD group and normal control group.The concentrations of serum GLP-1 among groups were compared with single factor analysis of variance , and the concentrations of serum GLP-1 between the two groups were compared using LSD-t test.Analysing the correlation between GLP-1 and other indicators with Pearson analysis.Results The fasting GLP-1 levels of LOAD group were ( 123.4 ±20.8 ) nmol/L, and they were highest between the normal control group (78.6 ±6.0) nmol/L and the cerebral blood vessel disease group(89.0 ±8.7)nmol/L (F values were 3.46 and 1.98, P0.05).Deficient secretion of GLP-1 after taking glucose 1 hour in most of the patients of T2DM combined with LOAD group (99.1 ±14.2) nmol/L, LOAD group(73.9 ±6.6 ) nmol/L and T2DM group (96.3 ±7.0 ) nmol/L could be concluded .The GLP-1 levels of T2DM combined with LOAD group after taking sugar 2 hour were (115.4 ±18.6)nmol/L ,and were higher than that of normal levels (63.3 ±6.2) nmol/L after taking sugar 2 hour(t=4.49,P0.05).Pearson correlation analysis showed that the relationship of the levels of GLP-1 with Aβ( 1-42 ) and the levels of GLP-1 after taking glucose 1 h and 2 h were positively relative, and its coefficients of correlation were 0.401,0.436,0.722.Conclusions LOAD and T2MD are similar, and they have GLP-1 secretion shortage phenomenon after taking glucose , so monitoring dynamic change of GLP-1 after taking glucose may contribute to the auxiliary diagnosis of LOAD.
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Objective To compare the difference of kidney function evaluated by using 3 different estimated glomerular filtration rate(GFR) equations in populations .Methods Retrospectively analyzed 65 856 patients who measured serum creatinine and Cysta‐tin C at the same time ,and come from the outpatients or inpatients of the hospital .The estimated GFR (eGFR) were calculated through 3 equations ,then compared the eGFR in the population and among different groups according to different kidney functions , and then grouped the people enrolled in the study again according to the eGFR calculated by using the 3 different equations and compared the differences among groups .Results Compared with the eGFR calculated by using Creatinine equation ,the correlation coefficients of the eGFRs calculated by using the other two equations were 0 .81 and 0 .90 ,respectively ,both P<0 .05 ;The differ‐ence between the means of eGFR were 6 .19 and 1 .79 mL/(min × 1 .73 m2 ) respectively with obvious significance (P<0 .01) ,in consistency analysis .There were obvious overestimation of kidney function when using Creatinine equation to calculate eGFR .Con‐clusion There is consistence and obvious difference by using the 3 CKD‐EPI′s eGFR equations .Physicians should choose suitable equations to evaluate kidney function in different populations .