Mononuclear Cells Of Children Research Articles

Expression of LINC00847 in Peripheral Blood Mononuclear Cells of Children with Asthma and Its Prediction between Asthma Exacerbation and Remission.

Objective Asthma is defined as a heterogeneous disease that is usually characterized by chronic airway inflammation. Long noncoding RNAs play important roles in various biological processes including inflammation. To know more about the relationships between lncRNAs and asthma, we sought to the role of LINC00847 in peripheral blood mononuclear cells (PBMCs) of children with asthma exacerbation or asthma remission. Methods Microarray analysis was performed on GSE143192 and GSE165934 datasets to screen differentially expressed lncRNAs (DElncRNAs) in human PBMCs between asthma patients and normal controls. LINC00847 was selected from DElncRNAs in human PBMCs between asthma patients and normal controls for further investigation. The expression levels of LINC00847 were quantified in PBMCs collected from 54 children with asthma exacerbation, 54 children with asthma remission, and 54 healthy children by real-time qPCR. The forced expiratory volume in the first second in percent predicted values (FEV1%), ratio of forced expiratory volume in 1 second to forced vital capacity (FEV1/FVC), and peak expiratory flow rate (PEF%) were tested for evaluation of lung function. The concentration of immunoglobulin E (IgE) and eosinophil count was examined. The serum levels of interleukin-4 (IL-4), interferon-γ (IFN-γ), and IL-17A were determined by the ELISA method. Results The expression level of LINC00847 in PBMCs of asthma exacerbation children was remarkably higher than that in PBMCs of asthma remission children and healthy children (p < 0.001); the expression level of LINC00847 in PBMCs of asthma remission children was notably higher than that in PBMCs of healthy children (p < 0.001). Pearson correlation analysis revealed that the expression levels of LINC00847 in PBMCs of asthma children were negatively correlated with FEV1% (r = −0.489), FEV1/FVC (r = −0.436), PEF% (r = −0.626), and IFN-γ level (r = −0.614) of asthma children, but positively correlated with IgE concentration (r = 0.680), eosinophil count (r = 0.780), IL-4 (r = 0.524), and IL-17A (r = 0.622) levels. When LINC00847 expression was used to distinguish asthma exacerbation from asthma remission, a 0.871 AUC (95% CI: 0.805–0.936) was yielded with sensitivity of 79.63% and specificity of 77.78%. Conclusion The study demonstrates that increased LINC00847 expression may be associated with the development and progression of asthma, possibly serving as a novel biomarker for predicting asthma exacerbation from asthma remission.

Limited induction of SARS-CoV-2-specific T cell responses in children with multisystem inflammatory syndrome compared with COVID-19.

Why multisystem inflammatory syndrome in children (MIS-C) develops after SARS-CoV-2 infection in a subset of children is unknown. We hypothesized that aberrant virus–specific T cell responses contribute to MIS-C pathogenesis. We quantified SARS-CoV-2–reactive T cells, serologic responses against major viral proteins, and cytokine responses from plasma and peripheral blood mononuclear cells in children with convalescent COVID-19, in children with acute MIS-C, and in healthy controls. Children with MIS-C had significantly lower virus-specific CD4+ and CD8+ T cell responses to major SARS-CoV-2 antigens compared with children convalescing from COVID-19. Furthermore, T cell responses in participants with MIS-C were similar to or lower than those in healthy controls. Serologic responses against spike receptor binding domain (RBD), full-length spike, and nucleocapsid were similar among convalescent COVID-19 and MIS-C, suggesting functional B cell responses. Cytokine profiling demonstrated predominant Th1 polarization of CD4+ T cells from children with convalescent COVID-19 and MIS-C, although cytokine production was reduced in MIS-C. Our findings support a role for constrained induction of anti–SARS-CoV-2–specific T cells in the pathogenesis of MIS-C.

Study of Hepatitis C Genotypes in Serum and Peripheral Blood Mononuclear Cells in Children with Chronic Hepatitis C Infection.

Study of Hepatitis C Genotypes in Serum and Peripheral Blood Mononuclear Cells in Children with Chronic Hepatitis C Infection.

Expression of Nod-like receptor protein 3 inflammasome in peripheral blood mononuclear cells of children with Kawasaki disease in the acute stage

To study the association of Nod-like receptor protein 3 (NLRP3) inflammasome with inflammatory response in the acute stage and coronary artery lesion (CAL) in children with Kawasaki disease (KD). A total of 42 children with KD who were hospitalized from January to October 2017 were enrolled as the KD group, among whom 9 had CAL (CAL group) and 33 had no CAL (NCAL group). Fifteen age- and gender-matched children with pneumonia and pyrexia were enrolled as the pneumonia-pyrexia group. Fifteen healthy children were enrolled as the healthy control group. Real-time PCR was used to measure the mRNA expression of NLRP3 inflammasome (NLRP3, ASC and caspase-1) in peripheral blood mononuclear cells. The Spearman rank correlation test was used to investigate the correlation of NLRP3 mRNA expression with serum levels of C-reactive protein, erythrocyte sedimentation rate, interleukin-6, interleukin-1β, procalcitonin, albumin and prealbumin. The KD group had significantly higher mRNA expression of NLRP3, ASC and caspase-1 in the acute stage than the pneumonia-pyrexia and healthy control groups (P<0.05). The CAL group had significantly higher mRNA expression of NLRP3 than the NCAL group (P<0.05). NLRP3 mRNA expression was correlated with C-reactive protein, interleukin-6, interleukin-1β, and prealbumin levels in children with KD in the acute stage (rs=0.449, 0.376, 0.427, and -0.416 respectively; P<0.05). NLRP3 inflammasome may participate in inflammatory response in the acute stage and the development of CAL in children with KD.

Low Mononuclear Cell IL-18 and IL-27 Response in Children: Susceptibility to Tuberculosis Infection after Contact

Background Identification of the immune response against tuberculosis is vital to develop new diagnostic and therapeutic modalities. The objective of this study was to determine IL (interleukin)-18 and IL-27 responses of peripheral blood mononuclear cells to early secreted antigen (ESAT-6) and culture filtrate protein-10 (CFP-10) stimulation in children with a (+) or (−) tuberculin skin test (TST) with in-house tuberculosis contact. Methods We enrolled 40 children aged 1 to 5 years who had an in-house contact with a tuberculous adult. Blood samples were obtained from all children for QuantiFERON tuberculosis (TB) gold in tube (QFT-GIT), and peripheral blood mononuclear blood cell culture tests. The subjects were grouped as TST (−) QFT-GIT (−), TST (+) QFT-GIT (−), and TST (+) QFT-GIT (+). Supernatant of peripheral blood mononuclear cell culture was separated with and without stimulation of ESAT-6 and CFP-10, and IL-18 and IL-27 levels were measured with enzyme linked immunoassay (ELISA) test. Results The study group included 22 boys and 18 girls with mean age 4.25 ± 0.9 years. IL-18 and IL-27 levels were statistically significant in ESAT-6/CFP-10-stimulated supernatants of peripheral blood mononuclear cell (PBMC) samples among the three groups (p = 0.000, p = 0.007, respectively). IL-18 levels between the TST (−) QFT-GIT (−) and TST (+) QFT-GIT (+) groups were significantly different (p = 0.026). Both IL-18 and IL-27 levels were significantly different between ESAT-6/CFP-10 stimulated PBMC supernatants of TST (−) QFT-GIT (−) and TST (+) QFT-GIT (−) groups (p = 0.000, p = 0.003, respectively). Conclusion Low IL-18 and IL-27 responses of peripheral blood mononuclear cells in children with Bacillus Calmette–Guérin (BCG) vaccine may play a role in Mycobacterium tuberculosis infection after in-house contact.

Expression and clinical significance of signal transducer and activator of transcription-1, interferon regulatory factor-1, and interferon-γ in children with bronchial asthma

Objective To study the expression profiles of signal transducer and activator of transcription-1(STAT1), interferon regulatory factor-1(IRF1), and interferon-γ(IFN-γ) in peripheral blood mononuclear cells in children with bronchial asthma, in order to further study their roles in the pathogenesis of asthma. Methods Thirty-eight children with acute exacerbation of bronchial asthma who visited pediatric outpatient clinic or hospitalized in pediatric department from September 2017 to May 2018 were enrolled as the asthma group.After standardized treatment, the 38 children with asthma were divided into the effective asthma treatment group(n=23) and the ineffective asthma treatment group(n=15), according to the treatment effect.Twenty-five healthy children were enrolled as the control group.RT-PCR and Western blot were used to measure the mRNA and protein expression levels of STAT1, IRF1, and IFN-γ in peripheral blood mononuclear cells.The correlations between the mRNA and protein expression of STAT1, IRF1, and IFN-γ were analyzed. Results The asthma group, the effective asthma treatment group and the ineffective asthma treatment group had significantly higher mRNA and protein expression levels of STAT1 and IRF1 than those of the control group (P<0.05). The ineffective asthma treatment group had significantly higher mRNA and protein expression levels of IRF1 than those of the effective asthma treatment group (P<0.05). The asthma group, the effective asthma treatment group and the ineffective asthma treatment group had significantly lower mRNA and protein expression levels of INF-γ than those of the control group (P<0.05). Conclusion The expression level of STAT1 increases in children with asthma, while the expression level of IFN-γ decreases in those children.The expression level of IRF1 increases in children with asthma and is closely related to the treatment effectiveness. Key words: Bronchial asthma; Signal transducer and activator of transcription-1; Interferon regulator factor-1; Interferon-γ; Children

Expressions of Toll like receptor-1, -3, -9 in peripheral blood mononuclear cells of children with idiopathic nephrotic syndrome

Objective To investigate the expressions of Toll-like receptor (TLR)-1, TLR-3, TLR-9 in peripheral blood mononuclear cells (PBMC) of children with idiopathic nephrotic syndrome (INS). Methods The expressions of TLR-1, TLR-3 and TLR-9 mRNA were measured by real time-polymerase chain reaction (RT-PCR) in PBMC of 45 children with INS came from the First Affiliated Hospital of Xinxiang Medical University before and after treatment (active group and remission group, respectively) and 30 healthy children from medical examination center as healthy control group, so as to explore the relativity between TLR-1, TLR-3 and TLR-9 mRNA and serum levels of albumin and cholesterol. Results The expression of TLR-1 mRNA/β-actin in PBMC of children with INS before treatment (the active group) was significantly higher than that in the remission group (2.432±0.231 vs.1.675±0.627, t=7.599, P 0.05). The correlation analysis showed that there was a negative correlation between the expressions of TLR-1, TLR-3 and TLR-9 mRNA and the serum level of albumin in PBMC of children with INS before treatment (r=-0.457, -0.891, -0.125, respectively, all P<0.05). However, there was a positive correlation between the expressions of TLR-1, TLR-3 and TLR-9 mRNA and the serum level of cholesterol (r=0.445, 0.911, 0.872, all P<0.05). Conclusions The expressions of TLR-1, TLR-3 and TLR-9 mRNA in PBMC of children with INS were significantly higher and correlated with activity of disease. Key words: Idiopathic nephrotic syndrome; Child; Toll-like receptor; Peripheral blood mononuclear cells

Doxycycline suppresses Chlamydia pneumoniae induced interferon-gamma responses in peripheral blood mononuclear cells inchildren with allergic asthma.

Persistent respiratory infections caused by Chlamydia pneumoniae have been implicated in the pathogenesis of chronic diseases (e.g. asthma). Antibiotics are used to treat C.pneumoniae respiratory infections; however, the use of antibiotics as anti-inflammatory agents in treatment of asthma remains controversial. The current study investigated whether ciprofloxacin, azithromycin, or doxycycline can suppress C.pneumoniae-induced production of immunoglobulin (Ig) E or cytokines in peripheral blood mononuclear cells (PBMC) obtained from asthmatic children. Apart from blood, nasopharyngeal swab specimens were also collected to test for the presence of C.pneumoniae and/or M.pneumoniae (qPCR). PBMC (1.5 x 106) from asthmatic pediatric patients (N=18) were infected or mock infected for 1h±C.pneumoniae AR-39at a multiplicity of infection (MOI)=0.1, and cultured±ciprofloxacin, azithromycin, or doxycycline (0.1 or 1.0μg/mLmL) for either 48h(cytokines) or 10 days (IgE). Interleukin (IL)-4, interferon (IFN)-γ and IgE levels in supernatants were measured (ELISA). When PBMC were infected with C.pneumoniae, IL-4 and IFNγ production increased (p=0.06 and 0.03, respectively); IgE levels were low. The now-elevated levels of IL-4 didn't decrease significantly after addition of ciprofloxacin, azithromycin, or doxycycline. However, infected PBMC IFNγ formation decreased significantly when 0.1μg/mL doxycycline was employed (p=0.04); no dose of ciprofloxacin or azithromycin had any impact. This inhibitory outcome with doxycycline lends support to the use of tetracyclines as immune modulators and anti-inflammatory medications in treatment of C.pneumoniae-infected asthma patients.

Effect of Levetiracetam add-on therapy on expression of multidrug resistant-associated protein 1 and 2 in peripheral blood mononuclear cells of children with intractable epilepsy

Objective To explore the changes of multidrug resistant-associated protein (MRP)1 expression and MRP2 expression in the peripheral blood mononuclear cells in intractable epileptic children after accepting the Levetiracetam (LEV) add-on therapy. Methods From June 2012 to September 2015, 68 intractable epileptic children who had been diagnosed at the Outpatient Department of the Xuzhou Children′s Hospital were randomly selected and given the LEV add-on therapy.The expressions of MRP1 and MRP2 in their peripheral blood mononuclear cells were detected by real-time fluorescence quantitative polymerase chain reaction and Western blot before the LEV add-on therapy and 6 months after the LEV add-on therapy. Results (1) Compared to the expressions of MRP1 mRNA and MRP2 mRNA in the peripheral blood mononuclear of intractable epileptic children before therapy, the expressions of MRP1 mRNA (0.23±0.02) and MRP2 mRNA (0.17±0.03) after therapy were obviously down-regulated with significant differences(tMRP1=32.77, P<0.01; tMRP2=26.73, P<0.01). (2)The relative expressions of protein of MRP1 and MRP2 in the peripheral blood mononuclear of intractable epileptic children before therapy were 0.59±0.03, 0.53±0.02 respectively, and they were 0.39±0.03, 0.38±0.02 after therapy.Compared with those before therapy, the protein expressions of MRP1 and MRP2 were down-regulated, and the differences were significant (tMRP1=11.55, P<0.01; tMRP2=9.87, P<0.01). Conclusions The expressions of MRP1 and MRP2 mRNA and protein in the peripheral blood mononuclear cells in some intractable epileptic children can be down-regulated if patients receive the LEV add-on therapy, and LEV may control the seizures of intractable epileptic children by down-regulating the expressions of MRP1 and MRP2, then reduce the occurrence and development of drug-resistant epilepsy. Key words: Intractable epilepsy; Levetiracetam; Multidrug resistant-associated protein; Child

Preparing for re-entry: is sequential switching the result of recurrent secondary responses?

Preparing for re-entry: is sequential switching the result of recurrent secondary responses?

Prevention of diabetes in overweight/obese children through a family based intervention program including supervised exercise (PREDIKID project): study protocol for a randomized controlled trial

BackgroundThe global pandemic of obesity has led to an increased risk for prediabetes and type-2 diabetes (T2D). The aims of the current project are: (1) to evaluate the effect of a 22-week family based intervention program, including supervised exercise, on insulin resistance syndrome (IRS) risk in children with a high risk of developing T2D and (2) to identify the profile of microRNA in circulating exosomes and in peripheral blood mononuclear cells in children with a high risk of developing T2D and its response to a multidisciplinary intervention program including exercise.MethodsA total of 84 children, aged 8–12 years, with a high risk of T2D will be included and randomly assigned to control (N = 42) or intervention (N = 42) groups. The control group will receive a family based lifestyle education and psycho-educational program (2 days/month), while the intervention group will attend the same lifestyle education and psycho-educational program plus the exercise program (3 days/week, 90 min per session including warm-up, moderate to vigorous aerobic activities, and strength exercises). The following measurements will be evaluated at baseline prior to randomization and after the intervention: fasting insulin, glucose and hemoglobin A1c; body composition (dual-energy X-ray absorptiometry); ectopic fat (magnetic resonance imaging); microRNA expression in circulating exosomes and in peripheral blood mononuclear cells (MiSeq; Illumina); cardiorespiratory fitness (cardiopulmonary exercise testing); dietary habits and physical activity (accelerometry).DiscussionPrevention and identification of children with a high risk of developing T2D could help to improve their cardiovascular health and to reduce the comorbidities associated with obesity.Trial registrationClinicalTrials.gov, ID: NCT03027726. Registered on 16 January 2017.

Chlamydia pneumoniae Induces Interferon Gamma Responses in Peripheral Blood Mononuclear Cells in Children with Allergic Asthma

Respiratory infections caused by Chlamydia pneumoniae have been associated with exacerbations of asthma. Cell-mediated immunity (CMI) is critical for maintaining immunity. We compared interferon (IFN)-γ responses in C. pneumoniae-infected peripheral blood mononuclear cells (PBMC) in paediatric patients ± asthma. Presence of C. pneumoniae was tested from asthma patients (N=17) and non-asthmatic controls (N=16) (PCR). PBMC were infected for 1h ± C. pneumoniae AR-39 (MOI=0.1) and cultured for 48h. IFN-γ levels were measured in supernatants (ELISA). C. pneumoniae-IgG antibodies in serum were determined (MIF). All subjects tested negative for C. pneumoniae (PCR). C. pneumoniae-induced IFN-γ production invitro was more prevalent in asthma compared with non-asthma; levels of IFN-γ were higher in asthma compared with non-asthma (P=0.003). There was no association between recent respiratory infection and positive IFN-γ responses. These data show that C. pneumoniae modulates IFN-γ responses in patients with asthma, even in absence of active infection.

Expression profiles of PI3K, NF-κB, and STAT1 in peripheral blood mononuclear cells in children with bronchial asthma

To study the expression profiles of PI3K, NF-κB, and STAT1 in peripheral blood mononuclear cells (PBMCs) in children with bronchial asthma, as well as their roles in the pathogenesis of asthma. Thirty children with acute exacerbation of bronchial asthma were enrolled as the asthma group, and 20 healthy children were enrolled as the control group. RT-PCR and Western blot were used to measure the mRNA and protein expression levels of PI3K, NF-κB, and STAT1 in PBMCs. A spirometer was used to compare the pulmonary function between the two groups. The correlations between the mRNA expression of PI3K, NF-κB, and STAT1 and pulmonary function in children with bronchial asthma were analyzed. The asthma group had significantly higher mRNA and protein expression levels of PI3K, NF-κB, and STAT1 than the control group (P<0.05). Compared with the control group, the asthma group showed significant reductions in pulmonary function indices such as FEV1%, FEV1/FVC, and PEF% (P<0.05). In children with bronchial asthma, the mRNA expression levels of PI3K, NF-κB, and STAT1 were negatively correlated with FEV1%, FEV1/FVC, and PEF% (P<0.05). The expression levels of PI3K, NF-κB, and STAT1 increase in children with asthma, and are negatively correlated with pulmonary function indices, suggesting that PI3K, NF-κB and STAT1 are involved in the development and progression of bronchial asthma in children.

Alterations in immunophenotype of peripheral blood mononuclear cells in children with chronic viral hepatitis following anti-viral therapy

The aim of this study was to investigate the impact interferon type I based anti-viral therapy on the immune system of chronically infected children with hepatitis C and B virus. Peripheral blood mononuclear cells were examined by three-color flow cytometry, using a panel of fluorochrome labeled monoclonal antibodies. It was found that the majority of white blood cell subsets were decreased in patients after termination of long-term anti-viral treatment. In particular, potentially cytotoxic, effector cell subsets, such as natural killer, natural killer T-lymphocytes and CD8+CD28- T cells were significantly decreased. Moreover, plasmacytoid dendritic cells were decreased, both in percentage and in absolute values. These data suggest that interferon-α based treatment, apart from its anti-viral effect, has also remarkable impact on patient's cell-mediated immunity.

Expression of EB virus in peripheral blood mononuclear cells in children with systemic lupus erythematosus

Objective To discuss the role of EB virus(EBV)in the pathogenesis of systemic lupus erythematosus(SLE) in children through investigating the copies of EBV DNA and expression of EBV genes in peripheral blood mononuclear cells(PBMCs). Methods (1)PBMCs were isolated from 30 patients with SLE and 12 healthy normal controls respectively and DNA was extracted from PBMCs.(2) PBMCs were co-cultured with EBV for 12 days and RNA was extracted from PBMCs.(3)Real-time fluorescence quantitative PCR(Real-time PCR) was applied to detect the copies of EBV DNA in PBMCs.(4)Reverse transcription PCR was applied to detect expression of EBV genes. Results (1) Compared with the healthy control group[(40.1±11.6) copies/μg], a significant increase of EBV DNA copies was observed in SLE group[(658.6±183.6) copies/μg](P 0.05). (3)After PBMCs got co-cultured with EBV, expression of latent EBV genes and lytic genes were both increased in the patients and healthy controls.The latent EBV genes including latent membrane protein 1(LMP1), LMP2, EBV nuclear antigen 1 and the lytic genes including BCRF1, BLLF1 were all increased significantly in the patients compared with the healthy controls(all P<0.05). Conclusions There is a significant increase of EBV DNA copies and aberrant expression of EBV genes in SLE patients, which suggests that EBV may contribute to the pathogenesis of SLE. Key words: Child; Lupus erythematosus, systemic; EB virus; Gene expression

Expression and effect of serum interleukin-24 level on bone marrow mononuclear cells in children with acute leukemia.

To investigate the expression of interleukin-24 (IL-24) in the children with acute leukemia (AL) and its effect on the apoptosis of bone marrow mononuclear cells (BMMNCs) in vitro. Four groups were assessed: acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), non-leukemia, and healthy groups, 20 children in each group. ELISA was used to measure IL-24 serum level. The bone marrow was taken from patients and controls. BMMNCs were isolated and the DNA was analyzed by glucose electrophoresis. Flow cytometry was used to determine BMMNC apoptosis. The serum level of IL-24 in the ALL and AML groups were significantly higher than in the other two groups. There was no statistical difference between ALL and AML groups, either between non-leukemia and healthy groups. BMMNCs were exposed to IL-24 for 48 h, and the apoptotic rate of the group treated with 50 ng/ml IL-24 was obviously higher than that of control group (0 ng/mL). The serum IL-24 level of AL children decreased comparing to non-leukemia and healthy children, indicating that IL-24 can induce BMMNCs apoptosis of AL children in vitro.

Significance of multidrug resistance associated protein 1 and protein 2 expressions in peripheral blood mononuclear cells in children with intractable epilepsy

Objective To study the expression of multidrug resistance associated protein 1 (MRP1) and MRP2 in the peripheral blood mononuclear cells of children with intractable epilepsy (IE). Methods During Nov.2010 to Oct.2013, 50 children with IE were collected as the experimental group, simultaneously 50 children with epilepsy controlled by the anti-epileptic drugs (AEDs) and 50 healthy children without epilepsy were collected as the control group from the outpatient or inpatient of Xuzhou Children's Hospital. The expressions of MRP1 and MRP2 in the peripheral blood mononuclear cells of children were detected by reverse transcriptase polymerase chain reaction and Wes-tern blot. Results 1.The mean relative expression of MRP1 and MRP2 mRNA in the peripheral blood mononuclear cells of children with IE (0.795±0.042, 0.804±0.023) were higher than those in epilepsy controlled by AEDs (0.682±0.030, 0.675±0.021)and healthy children without epilepsy (0.665±0.031, 0.654±0.029) (all P 0.05). Conclusions MRP1 and MRP2 over-expression in the peripheral blood mononuclear cells of children with IE may be associated with drug-resistance mechanism for medically intractable epilepsy. Key words: Intractable epilepsy; Multidrug resistant associated protein; Peripheral blood mononuclear cell; Child

Bioinformatics Analysis of Gene Expression in Peripheral Blood Mononuclear Cells from Children with Type 1 Diabetes in 3 Periods

To explore the expression changes of potential key genes and relevant biological processes in peripheral blood mononuclear cells of children with newly diagnosis of type 1 diabetes (T1D).Microarray data GSE9006 were downloaded from Gene Expression Omnibus (GEO) database, including peripheral blood mononuclear cells samples from 43 children with newly diagnosed T1D (NEW), 19 one-month (1-MO) follow-up samples, 19 4-month (4-MO) follow-up samples and 24 healthy controls. The differentially expressed genes (DEGs) were identified using Affy package in R, and cluster analysis of DEGs were performed following functional enrichment analysis with Database for Annotation, Visualization and Integrated Discovery (DAVID) and construction of protein-protein interaction (PPI) network with STRING database.We identified 73, 73, 96 DEGs in NEW group, 1-MO group and 4-MO group, respectively by comparing with healthy controls with |logFC|>0.58 and P-value<0.05. The cluster analysis of these DEGs showed that 4 genes, including human leukocyte antigen (HLA-DQA1), HLA-DRB4, integrin 3 (ITGB3) and killer cell lectin-like receptor subfamily F member 1 (KLRF1) were all significantly expressed in 3 groups, which were significantly enriched in asthma, T1D and intestinal immune network for IgA production pathway. And 57 genes enriched in cluster 5, which were only differentially expressed in NEW group, were involved in response to wounding, inflammatory response and blood coagulation as well as chemokine signaling pathway. Besides, the hub genes in PPI network of cluster 5 were identified, containing FOS, pro-platelet basic protein (PPBP), interleukin 8 (IL8), formyl peptide receptor-like 2 (FPR2) and platelet factor 4 (PF4).HLA-DQA1, HLA-DRB4, ITGB3 and KLRF1 might be targets for treatment of T1D, and 5 hub proteins, FOS, PPBP, IL8, FPR2 and PF4, were likely to be new markers for diagnosis of T1D.

Profiles of innate immune receptors and cytokines in peripheral blood mononuclear cells in children with measles

The aim of this study was to explore the roles and clinical significance of innate immune receptors and cytokine in children with measles. The children with measles hospitalized in the department of infectious diseases, Children's Hospital of Fudan University during 2009-2011 were enrolled into measles group, while the healthy children examined in well baby clinic were enrolled into control group. The mRNA expression of TLR2/3/4/7, melanoma differentiation-associated gene-5 (MDA-5), retinoic acid-inducible gene I (RIG-I), IFN-α/β and IL-10 in peripheral blood mononuclear cells were detected by real-time PCR. The protein levels of IFN-α, IFN-β and IL-10 in plasma were measured using ELISA. SPSS 13.0 software was applied to analyze the difference between two groups. Data from a total of 98 patients in measles group and 59 children in control group were collected. The mRNA expressions of TLR2, MDA-5 and RIG-I had no statistical significance between two groups (P > 0.05, respectively). The relative mRNA expressions of TLR3, TLR4, TLR7 in measles group (2.25 ± 0.74, 2.05 ± 0.72, 2.12 ± 0.29) were significantly lower than those in control group (2.09 ± 0.78, 1.90 ± 0.75, 1.87 ± 0.68) (P < 0.01; respectively). Both IFN-α and IFN-β had significantly decreased mRNA expressions in measles patients (2.41 ± 1.31, 2.47 ± 1.26) compared with those in controls (2.22 ± 0.48, 2.35 ± 0.64)(P < 0.01 respectively); however, IL-10 mRNA levels significantly increased (2.49 ± 0.58 vs. 2.62 ± 0.95) (P < 0.001). The IL-10 levels in plasma in measles group were significantly higher during the whole period of fever [<5 d group: 29.89 (25.82-38.15) ng/L and ≥ 5 d group:34.55 (28.26-38.70) ng/L] than that in control group [25.15 (24.20-27.38) ng/L] (P < 0.05 respectively). TLR3/4/7 mRNA expression was low in peripheral blood mononuclear cells of measles patients. Levels of IL-10 were significantly raised in the early stage after infection and lasted for a long time, and reduced IFN-α levels in plasma were associated with the fever durations of measles patients. These results indicated that multiple TLRs and cytokines may participate in the immune response after measles virus infection.

Effect of Cardiopulmonary Bypass on Annexin A1 Expression in Peripheral Blood Mononuclear Cells of Children with Congenital Heart Disease

Effect of Cardiopulmonary Bypass on Annexin A1 Expression in Peripheral Blood Mononuclear Cells of Children with Congenital Heart DiseaseThis study aimed to investigate the effect of cardiopulmonary bypass (CPB) on Annexin A1 expression in the peripheral blood mononuclear cells (PBMCs) of children with congenital heart disease (CHD). A total of 30 children receiving CPB for interventricular septal defect were included. Peripheral blood was collected before and after CPB. PBMCs were collected by density gradient centrifugation. Protein extraction was performed by lysis and subjected to 2D-QUANT for protein quantitation. Isoelectric focusing electrophoresis (IEF) was carried out followed by gel image analysis. Protein spots with a difference in expression of &gt;1.5 fold were collected as candidate proteins which were subjected to mass spectrometry for the identification of differentially expressed proteins. Western blot assay was employed to confirm the expressions of target proteins. Peripheral blood collected at two time points was subjected to two-dimensional electrophoresis, and a total of 12 differentially expressed proteins were identified. Of them, 5 proteins had decreased expression before CPB (T0) but their expressions increased after CPB (T1); the remaining 7 proteins had increased expressions before CPB but their expressions reduced after CPB. One of these differentially expressed proteins was Annexin A1. Western blot assay confirmed that Annexin A1 expression began to increase at 0.5 h after CPB, and the increase of Annexin A1 was more obvious after CPB. Our findings primarily indicate the potential mechanism underlying the role of PBMC in inflammatory response following CPB, and provide a target for the prevention and control of post-CPB systemic inflammatory response syndrome (SIRS).