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Objective:To investigate the clinical characteristics of the disease spectrum of pediatric hyper blood immunoglobulin E (IgE).Methods:A total of 498 children with total serum IgE ≥ 5×10 5 IU/L admitted to the Department of Pediatrics, Sun Yat-Sen Memorial Hospital of Sun Yat-Sen University from January 2012 to December 2018 were enrolled.Their clinical data, etiology distribution, clinical manifestations, laboratory results, treatment and outcomes were retrospectively analyzed.According to serum total IgE level, patients were divided into mildly increased IgE group (5×10 5-<10×10 5 IU/L), moderately increased group (10×10 5-<20×10 5 IU/L), and severely increased group (≥20×10 5 IU/L). The distribution of disease types among the 3 groups were compared. Results:(1) Allergic disease (213 cases) was the most common etiology in children with hyper blood IgE, and infectious disease (163 cases), mycoplasma pneumoniae (109 cases) and EB virus (120 cases) were common pathogens.(2) The incidence of allergic diseases (45.0%) and infectious diseases (42.2%) in the mildly increased group was significantly higher than that in the moderately increased group (40.8%, 26.2%, respectively) and the severely increased group(38.9%, 12.2%, respectively) (all P<0.001). The incidence of immune diseases(18.5%), tumors and hematological diseases (5.4%) in the moderately increased group was significantly higher than that of the mildly increased group (4.4%, 2.0%, respectively) (all P<0.001). The incidence of immune diseases (34.4%), tumors and hematological diseases (11.1%) in the severely increased group was significantly higher than that of the mildly increased group(4.4%, 2.0%, respectively) and the moderately increased group (18.5%, 5.4%, respectively) (all P<0.001). (3) The main clinical manifestations were fever (63.5%), respiratory symptoms (53.7%) and lympha-denopathy (53.7%), 47.5% of the children with hyper blood IgE had an increased white blood cell count, and 12.1% of them had an increased eosinophil count.(4) The most common specific allergens were dust mite combination (32.0%), milk (17.0%), and egg white (16.0%). There was no difference in disease distribution among the 3 groups of hyper blood IgE children with positive specific IgE ( P=0.164). Conclusions:Hyper blood IgE in children are most commonly caused by allergic and infectious diseases.The etiological distributions of hyper blood IgE in children at varying severities differ a lot.The higher the total IgE level, the higher the incidence of immunodeficiency disease, rheumatic disease, tumor and hematological disease.
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[Objective]To investigate the pharmacological mechanism of montelukast in the inhibition of inflammation.[Meth-ods]Respiratory syncytial virus-infected human bronchial epithelial cell(16HBEC)inflammatory cell model was established,and mRNA and protein expressions of Nuclear factor NF-E2 related factor(Nrf2),heme oxygenase(HO-1),quinone oxidoreductase (NQO-1),and glutathione transferase (GST) were determined by qPCR and Western-blot ,and production of cellular reactive oxygen species(ROS)was measured by DCFH-DA fluorescent probe method. Nrf2 siRNA was further synthesized to reduce the expression of Nrf2 ,to investigate the chang of inflammatory index.[Results]Montelukast significantly reduced the expressions of inflammatory cytokines IL-6,TNF-α,and IL-1β(P<0.05)on respiratory syncytial virus-infected 16HBEC,and the ROS level in inflammatory cell model was decreased(P<0.05),increased the mRNA and protein expressions of Nrf2,HO-1,NQO-1 and GST (P < 0.05),with a more significant effect at higher dose. After the down-regulation of Nrf2,the expressions of inflammatory cyto-kines IL-6,TNF-α,and IL-1βwere increased(P<0.05),and ROS level was significantly increased(P<0.05),mRNA and pro-tein expressions of Nrf2,HO-1,NQO-1 and GST were decreased(P<0.05).[Conclusion]Montelukast inhibits the inflammation of human bronchial epithelial cells infected by respiratory syncytial virus (RSV),and the potential mechanism may involve its ef-fect on the Nrf2/ARE signaling pathway.
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Objective To investigate the expression and clinical significance of miR-125b in patients before and after treatment of acute myeloid leukemia (AML) with different types.Methods The levels of miR-125b were measured by using real-time fluorescence quantification PCR (qRT-PCR) in 65 AML patients (all AML samples from the Sun Yat-Sen Memorial Hospital of Sun Yat-Sen University,and the control samples isolated from cord blood which were obtained after normal full-term delivery of babies) before and after treatment,and then the relationship between the levels of miR-125b and patients＇ sex,age,peripheral blood cells,type clinically,relapse and therapeutic effect were analyzed.Results Of newly diagnosed AML patients and the control samples,miR-125b positive rate was 100％.The miR-125b expression levels in the control group was 1.50,and its expression in AML was 11.06,and the difference was significant (P =0.036 6).In complete remission (CR) group of acute promyelocytic leukemia (M3),the expression of miR-125b after induction therapy was significantly reduced,and CR rate of miR-125b decreased group was 91.3％,while that of the increased group was 50.0％,and the difference was significant (P =0.042 6).The miR-125 b expression level was decreased from 29.7 ± 4.9 to 19.2 ± 6.0 after chemotherapy,and the difference was significant (P ＜ 0.036 6).In non-M3 AML,CR rate of miR-125 b decreased group was 86.7 ％,while that of the increase group was 42.1％,and the difference was significant (P =0.021 5).There was no correlation between miR-125b expression and patients＇ gender,age and peripheral blood cells.Conclusions The differences in expression of miR-125b is very important in disease occurrence and progress.Using qRT-PCR to dynamically detect the expression of miR-125b dynamically in AML patients before and after therapy may predict outcome more precisely and has the potentials as an effective biomarker in determining prognosis,monitoring the risk of recurrence,and guiding the treatment.
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AIM: To investigate the regulatory function of bone marrow-derived mesenchymal stem cells (MSCs) on T helper 17 cells (Th17) and regulatory T cells (Treg) in peripheral blood of severe asthmatic children . METHODS:MSCs were isolated , cultured and identified in vitro.MSCs digested with mitomycin were cocultured with T lymphocytes (TLC) at different ratios (1∶1, 1∶2, 1∶10 and 1∶20) from severe asthmatic children for 72 h.The prolifera-tion of TLC was measured by CCK-8 method.In the coculture system of the 1∶2 ratio and the single TLC system , the super-natant levels of interleukin-17 (IL-17) and transforming growth factor-β(TGF-β) were measured by ELISA.The mRNA expression of retinoic acid-related orphan nuclear receptor C (RORC) and forkhead box protein 3 (Foxp3) in TLC was de-tected by qRT-PCR.RESULTS:After cocultured with MSCs , the proliferation of TLC decreased significantly in a dose-dependent manner (P0.05).CONCLUSION: MSCs suppresses Th17 polarization of naive peripheral blood CD 4 +T cells and matures Th17 cells secreting IL-17, which may ef-fectively revise Th17/Treg imbalance of asthma .
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AIM:To investigate whether early endothelial progenitor cells (early-EPCs) expressβ2-adrenergic receptor (β2 AR) in the chronic obstructive pulmonary disease ( COPD) patients and the effect of β2 AR expression on the migration of early-EPCs.METHODS:Venous blood samples (20 mL) were obtained from antecubital vein of COPD pa-tients or healthy controls .Peripheral blood mononuclear cells were isolated by standard Ficoll gradient centrifugation , and purified by CD34 positive selection cocktail .The mRNA expression of β2 AR in the early-EPCs was detected by RT-PCR. The protein levels of β2 AR were assessed by Western blotting and flow cytometry .Chemotaxis was studied by Transwell as-say.Cultured early-EPCs were treated with ICI118551, norpinephrine (NE) or monoclonal antibody of β2AR (mAb-β2 AR) for 24 h.The number of migratory cells was counted under a light microscope .RESULTS:The level of β2 AR ex-pression in the COPD patients was higher than that in the controls .The number of migratory early-EPCs to stromal cell-de-rived factor 1αwas significantly improved by ICI 118551 compared with other COPD groups .When early-EPCs from the COPD patients or the controls were treated with different concentrations of mAb-β2 AR for 24 h, the number of migratory early-EPCs from the COPD patients and the controls treated with NE at concentration of 100 nmol/L was significantly re-duced.However, a marked decrease in the number of migratory early-EPCs from the COPD patients treated with NE was observed compared with control group .Before treated with ICI118551 or NE for 24 h, the early-EPCs were co-incubated with mAb-β2 AR for 40 min, and the number of migratory early-EPCs was not significantly different between COPD group and control group .Genetic down-regulation of β2 AR promoted the migration of early-EPCs in COPD group .CONCLU-SION:The level of β2 AR expression in the COPD patients is increased compared with the controls .The down-regulation ofβ2 AR improves the migration of early-EPCs.
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[Objective] This study was designed to determine Th1, Th2 cell numbers and investigate T-bet mRNA, GATA-3 mRNA expression of spleen MNC in a mufine asthmatic model which intended to understand effect of airway T-bet plasmid gene transfer on Th differentiation. [ Methods] A mouse asthmatic model was established by sensitization with ovalbumin (OVA). Thirty-two C57BL/6 mice were divided into four groups (8 mice in each group): the normal control group (group A ), the asthmatic model group (group B), the pcDNA3 plasmid group (group C), the pcDNA3-T-bet group (group D). All animals were sensitized and challenged with OVA, except group A normal saline was applied. The group C was intranasally administered 50 μg pcDNA3 plasmid at 24 h before intranasal challenges, and the 50 μg pcDNA3-T-bet plasmid for the mice of group D. We investigated Th1 and Th2 cell numbers by FACS and T-bet, GATA-3mRNA expression of spleen mononuclear cells (MNC) by semi-quantitative PCR in the four groups. [Result] Th1 percent in spleen MNC of pcDNA3-T-bet treated mice was significantly increased ([2.29±1.551% vs. [1.93±1.141%, P<0.05), while Th2 percent was significantly decreased ([0.93±0.64]% vs. [1.63±0.59]%), compared with that of the asthmatic control group mice by FACS. Spleen MNC was detected a high level of T-bet mRNA expression (0.53±0.027 vs. 0.28±0.035, P<0.05) and a low level of GATA-3 mRNA expression (0.24±0.022 vs. 0.58±0.038, P<0.05) after pcDNA3-T-bet treatment by RT-PCR. There was no significant change between the pcDNA3 plasmid group and the asthmatic model group. [Conclusion] The intranasal transfer of pcDNA3-T-bet plasmid was effective in modulating the imbalance of Th1/Th2 in mice asthma model, which provides a novel therapeutic strategy for transferring transcriptional factor in allergic asthma.
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[Objective] To investigate the changes of CD4~+ CD25~+ regulatory T cells (Tr) in peripheral blood and their relation with their body mass index (BMI) of children with acute attack asthma. [Methods] Peripheral blood was obtained from 70 children with acute attack asthma, 30 remission children, and 50 normal control children. Then 70 children with acute attack asthma, were divided by normal weight group (40 cases) and overweight group (30 cases). The levels of CD4~+CD25~+Tr of the patients were tested by flow cytometry (FCM), and their BMI were calculated. [ Results] The levels of CD4~+ CD25~+ Tr [(6.17± 1.72)%] in acute attack group were lower than that in remission group [(7.56±1.48)%] or that in the control group [(7.13± 1.48)%] (P<0.05), but no difference between that in the remission and that in the control (P>0.05). The CD4~+CD25~+Tr of asthmatic children with normal weight [(6.34±1.71)%] was higher than that of asthmatic children with overweight [(4.74±1.20)%] (P<0.05). There was a remarkably negative correlation between the level of CD4~+ CD25~+ Tr of asthmatic children [(6.17±1.72)%] and the BMI (16.00±2.14) (r_p=-0.814, P<0.05). [Conclusion] The levels of CD4~+ CD25~+Tr are remarkable decrease in attack asthmatic children, and more decrease in overweight patients. There is remarkably negative correlation between the levels of CD4~+CD25~+ Tr in peripheral blood of attack asthmatic children and their BMI.
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AIM: To investigate the effect of glucocorticoid inhalation on the levels of CD4~+CD25~+ regulatory T cells in peripheral blood of asthmatic children. METHODS: Glucocorticoid inhalator was inhaled by 70 children with attack asthma. The levels of CD4~+CD25~+ Tr in peripheral blood of asthmatic children were tested by flow cytometry (FCM). RESULTS: The CD4~+CD25~+ Tr levels in peripheral blood of asthmatic children were (5.62% ± 1.29% ) and (7.05% ± 1.61%) before and after of regulated glucocorticoid inhalation, respectively (P<0.01). The Tr levels were (7.56% ± 1.88% ) , (7.09% ± 1.23% ) and (6.11% ± 1.96% ) in the complete control group, part control group and poor control group, respectively ( P < 0.05 ). The Tr level in formal treatment group (7.05% ±1.61%) was higher than that in irregular treatment group ( 5.91 % ± 1.76% ), P < 0.01.CONCLUSION: The level of CD4~+CD25~+ Tr is remarkable increased by regulated glucocorticoid inhalation, and the level of Tr can reflect the effects of glucocorticoid inhalation.
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AIM: To investigate the effect of T-bet plasmid gene transfer to airway on allergen induced airway inflammation in a murine asthmatic model. METHODS: A mouse asthma model was established by sensitization with ovalbumin (OVA). Forty C57BL/6 mice were divided into 4 groups (10 mice in each group): the normal control group (group A), the asthmatic model group (group B), the pcDNA3 plasmid group (group C), and the pcDNA3-T-bet group (group D). The animals in group B, C and D were sensitized and challenged with OVA. The animals in group A were applied with normal saline. pcDNA3 plasmid at dose of 50 μg was intranasally administered at 24 h before intranasal challenges to the mice in group C, and the 50 μg pcDNA3-T-bet plasmid for the mice in group D. Bronchial alveolar lavage fluid (BALF) was collected and lung tissues were resected at 48 h after OVA challenge for later assay. RESULTS: After administration with pcDNA3-T-bet plasmid, high level of T-bet expression at 48 h was detected in the lung tissue by Western blotting. In pcDNA3-T-bet treated asthmatic models, histological evaluation revealed the significant suppression of eosinophil peribronchial and perivascular infiltration, and reduction of epithelial damage. The numbers of eosinophils, neutrophils and lymphocytes in BALF from pcDNA3-T-bet treated mice were significantly reduced compared to those in asthmatic control group (P<0.05). The level of IL-4 in BALF was significantly decreased in pcDNA3-T-bet group compared to that in asthmatic control group (P<0.05), while the level of IFN-γ in BALF was significantly increased in pcDNA3-T-bet group. No significant change of inflammation cells and cytokines in pcDNA3 plasmid group and asthmatic control group was observed (P>0.05). CONCLUSION: Intranasal pcDNA3-T-bet plasmid transfer inhibits asthmatic airway inflammation in the murine asthmatic model, suggesting a new therapeutic strategy for allergic asthma.
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In this study,IL-4 and IL-12 levels in plasma and peripheral blood mononuclear cells(PBMC) as well as serum IgE level were prospectively assessed with double-antibody sandwich ELISA technique in children with asthma during the attack group (30 cases)and the interval group(12 cases).The results observed revealed that serum IgE level and IL-4 level in plasma and PBMC after PHA and LPS provocation during both of the attack stage and the interval stage were found to be evidently higher than those of the normal control group (P＜0.01).Otherwise,IL-12 level during both stages was much lower than that of the normal control(P＜0.01).In other hand,there were found to have a significant difference in all these 3 data between attack and interval stages.Thus, the conclusion indicates that there might be an imbalance of IL-4,IL-12 and IgE level in children with asthma during both of the attack and the interval stages.
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AIM and METHODS: To study the immunological effect of measles vaccine therapy on asthmatic children, we examined the changes of interleukin-12 , interleukin-13 and total serum IgE levels in plasma and cultured peripheral blood mononuclear cells(PBMC) supernatant by means of ELISA in 13 mild-moderate asthmatic children treated with measles vaccine. Results were compared with 12 anti-symptomatic treatment mild-moderate asthmatic children and 17 normal children control group. RESULTS:After measles vaccine treatment, IL-13 and total serum IgE levels decreased remarkably, statistically lower than that of group receiving only anti-symptomatic treatment. There was no statistical difference in IL-12 level between the two group. Correlation analysis: 1)IL-12 level of plasma was negatively correlated to the level of serum total IgE, there was no correlation of supernatant IL-12 in PBMC to the total serum IgE; 2)IL-13 levels in plasma and PBMC were positively correlated to the level of total serum IgE; 3) IL-12 level was negatively correlated to IL-13. CONCLUSION: Measles vaccine could down-regulate IL-13 level, therefore decrease total IgE synthesis, but not affect IL-12 level in asthmatic children.