Role of hypomethylation of suppressor of cytokine signaling in T helper 17 cell/regulatory T cell imbalance in children with Henoch-Schönlein purpura / 中国当代儿科杂志

OBJECTIVE@#To investigate the association between suppressor of cytokine signaling (SOCS) hypomethylation and T helper 17 (Th17) cell/regulatory T (Treg) cell imbalance in children with Henoch-Schönlein purpura (HSP) and the immune pathogenesis of HSP.@*METHODS@#A total of 32 children in the acute stage of HSP who were hospitalized from May 2014 to January 2015 were enrolled as subjects, and 28 children who underwent physical examination were enrolled as normal control group. ELISA was used to measure the plasma level of interleukin-6 (IL-6). Flow cytometry was used to measure the percentages of CD4 IL-17A T cells (Th17 cells) and CD4CD25 Treg cells (Treg cells) in peripheral blood and mean fluorescence intensity (MFI) for phosphorylated-STAT3 (pSTAT3) protein in CD4 T cells. Quantitative real-time PCR was used to measure the mRNA expression of suppressor of cytokine signaling-1 (SOCS1) and suppressor of cytokine signaling-3 (SOCS3) in CD4 T cells. High-resolution melting (HRM) was used to evaluate the methylation level of the CpG islands in SOCS1 exon 2 and the CpG islands of the potential bind sites for STAT3 in the 5'-untranslated region (5'-UTR) of SOCS3 in peripheral blood mononucleated cells.@*RESULTS@#Compared with the normal control group, the HSP group had significant increases in plasma IL-6 concentration and MFI for pSTAT3 in CD4 T cells, as well as a significant increase in the percentage of Th17 cells and a significant reduction in the percentage of Treg cells (P<0.05). The HSP group had significantly higher mRNA expression of SOCS1 and SOCS3 in peripheral blood mononucleated cells than the normal control group (P<0.05). In the HSP group, the mRNA expression of SOCS1 and SOCS3 was negatively correlated with Th17/Treg ratio (P<0.05). The HSP group had hypomethylation of the CpG islands in SOCS1 exon 2 and the potential binding site for STAT3 in SOCS3 5'-UTR, while the normal control group had complete demethylation.@*CONCLUSIONS@#Low relative expression of SOCS1 and SOCS3 caused by hypomethylation may be a factor for Th17/Treg imbalance in children with HSP.

Effects of Astragalus membranaceus on cytokine secretion of peripheral dendritic cells in children with Henoch-Schonlein purpura in the acute phase / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effects of Astragalus membranaceus (AM) on the cytokines secretion of peripheral dendritic cells (DC), including interleukin-10, -12, and -18 (IL-10, IL-12 and IL-18), in children with Henoch-Schonlein purpura (HSP) in the acute phase; and to study the immunological regulation mechanism of AM.</p><p><b>METHODS</b>Peripheral blood mononuclear cells (PBMC) were obtained from 28 children with acute HSP by density gradient centrifugation, and each sample was divided into two parts, one untreated and one treated with AM. All cells were developed to mature DC through treating with recombinant human granulocyte macrophage colony stimulating factor (GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-alpha). Expression of CD83 in the surface of mature DC was detected by flow cytometry, and levels of IL-10, IL-12 and IL-18 in the supernatant were measured by ELISA.</p><p><b>RESULTS</b>The supernatant level of IL-12 was higher [(141.58 +/- 100.19) ng/L vs (96.18 +/- 76.65) ng/L, t = 3.90, P<0.01], while levels of IL-10 and IL-18 were lower (t = 2.70, P<0.05; t = 4.07, P<0.01) in AM treated PBMCs than those in the untreated ones.</p><p><b>CONCLUSION</b>AM can correct the immunologic dysfunction of HSP children through increasing the IL-12, and decreasing the IL-10 and IL-18 secretions of PBMCs.</p>

Levels of IL-12 produced by dendritic cells and changes of TH1/TH2 balance in children with Henoch-Schonlein purpura / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the changes of IL-12 produced by dendritic cells in peripheral blood in children with Henoch-Schonlein purpura (HSP), and to explore its influence on TH1/TH2 balance in order to elucidate its significance in the pathogenesis of HSP.</p><p><b>METHODS</b>The levels of interferon-gamma (IFN-gamma), interleukin-4 (IL-4) and interleukin-12 (IL-12) in plasma were determined by ELISA in 60 HSP children (HSP group) and 21 healthy children (Control group). Peripheral blood mononuclear cells (PBMC) of 22 HSP patients and 21 healthy children were cultured in vitro and then were transformed into dendritic cells. The levels of IL-12 in the supernatant were detected by ELISA and the positive expression rate of CD1a(+) was detected by indirect immunofluorescence procedure.</p><p><b>RESULTS</b>1) The levels of IFN-gamma and the ratio of IFN-gamma/IL-4 in plasma of the HSP group were lower than those of the Control group (IFN-gamma 30.59 +/- 11.27 pg/mL vs 43.38 +/- 19.19 pg/mL; IFN-gamma/IL-4 ratio 0.70 +/- 0.28 vs 1.33 +/- 0.57) (P < 0.01). The levels of IL-12 in the HSP group were also lower than those of the Control group (153.95 +/- 91.88 pg/mL vs 323.06 +/- 162.34 pg/mL; P < 0.01). In contrast, the levels of IL-4 were higher than those of the Control group (45.08 +/- 9.19 pg/mL vs 32.95 +/- 7.10 pg/mL; P < 0.01). The plasma levels of IL-12 positively correlated with the IFN-gamma levels (r=0.52, P < 0.01) and the ratio of IFN-gamma/IL-4 (r=0.43, P < 0.01) in the HSP group. 2) The IL-12 levels in the supernatant of the HSP group were lower than those of the Control group (357.06 +/- 153.56 pg/mL vs 489.80 +/- 213.45 pg/mL; P < 0.05), and had a positive correlation with the plasma IL-12 levels (r=0.74, P < 0.01). 3) The positive expression rate of CD1a(+) of the HSP group was lower than that of the Control group [(27.42 +/- 10.75)% vs (35.68 +/- 12.18)%; P < 0.05], and positively correlated with the IL-12 levels in the supernatants (r=0.57, P < 0.01) and in plasma (r=0.68, P < 0.01).</p><p><b>CONCLUSIONS</b>There was an imbalance of TH1/TH2 in HSP children. The decrease of TH1 function had a positive correlation with the low levels of IL-12 in plasma, while the latter correlated closely with decreased number and / or function of dendritic cells, suggesting that the decreased number and / or function of dendritic cells in peripheral blood resulted in the imbalance of TH1/TH2 indirectly.</p>

Changes of Soluble Tumor Necrosis Factor-? Receptor Type 1 in Cerebrospinal Fluid of Children with Virus Encephalitis and Its Clinical Significance / 实用儿科临床杂志

Objective To explore the changes of soluble tumor necrosis factor receptor type 1(sTNFR1)in children with acute virus encephalitis(VE)and its clinical significance.Methods The levels of tumor necrosis factor-?(TNF-?),sTNFR1 and neuron specific enolase(NSE)in cerebrospinal fluid were determined by enzyme-linked immunosorbent assay(ELISA)in 55 children who were admitted with VE,including 25 cases with severe VE(SVE)and 30 cases with mild VE(MVE).Fifteen cases without VE were as control group.Results The levels of TNF-?,sTNFR1 in cerebrospinal fluid in encephalitis children were significantly higher than that in control group(Pa

Influence of Azithromycin on Helper T Lymphocyte Cell in Peripheral Blood of Children with Bronchitic Asthma / 实用儿科临床杂志

Objective To explore the influence of Azithromycin on helper T lymphocyte cell(Th)1/Th2 in peripheral blood of children with bronchitic asthma.Methods Twenty-four asthmatic children and 20 healthy children were selected.Peripheral blood mononuclear cells (PBMC) were isolated from venous blood and made into cells suspension in aseptic condition.0.2 mg/L,0.1 mg/L,0.05 mg/L and 0 mg/L Azithromycin were added into the cultures in asthmatic group.The control group was not interfered with Azithromycin.The supernatant was collected after 48 h.The levels of IFN-?,IL-4 and IL-10 in the supernatant were determined by enzyme-linked immunosobent assay(ELISA).SPSS 11.5 software was used to analyze data.Results 1.The level of IL-4 produced from PBMC of asthmatic group was significantly higher than that of control group(P0.05).2.Azithromycin 0.1 mg/L more promoted the secretion of IL-4 than the other 3 concentrations(Pa0.05).3.Azithromycin 0.2 mg/L and 0.1 mg/L more increased the level of IL-10 than the control group(P0.05).Conclusions The routine drug level of Azithromycin(0.1 mg/L) had no effects on the imbalance of Th1/Th2 of asthmatic children,but could modulate the immunological function by up-regulating the level of IL-10.

Cerebrespinal fluid ferritin in the child patients with acute lymphatic leukemia / 实用儿科临床杂志

Objcetives To sparch for the change of cerebrospinal fluid ferritin (CSF-Ft) content and its clinical significance.Methods The 42 patients with acute lymphatic leukemia(ALL)diagnosed by bone marrow biopsy were divided into 3 groups. There were 14 cases in Ⅰ group [introduced therapy stage without central nervous system leukemia(CNSL)7, 24 cases in Ⅱgroup (complete remission stage without CNSL) and 18 cases in Ⅲ grotip (with CNSL). There were 17 patients with viral encephalitis in viral encephalitis group and 15 patients without central nervous system oisease in control group. The CSF-Ft and SFt were determined by radioimmunoassay.Results The CSF-Ft contents of Ⅰ、Ⅱ、Ⅲ 、viral encepbalitis and control groups are 7.03 ?2.21 ?g/L,6.75 ?1.94?g/L, 31.06 ? 8.85?g/L, 7.26?1.83?g/L and 6.52 ?1.57?g/L. The CSF-Ft content in Ⅲ group are bigher than that in the other group (P

The Changes of T Cells Subpopulations and Immunoglobulin and Their Relationship in Children Patients with Simple Nephrotic Syndrome / 实用儿科临床杂志

Objective To search for the changes of T cells subpopulations and immunoglobulin and their relation-ship in children patients with simple nephrotic syndrome. Design Case-control research. Patients aud Participants 39 patients with simple nephrotic syndrome were divided into two groups:the incipient group and relapse group (6 cases were determined at the incipient and relapse time) .Thereare 28 patients in incipient group, 19 males and 9 females, at the age of 2 to 10 years old. There are20 patients in relapse group, 12 males and 8 females, at the age of 3 to 13 years old. There are 35health children in control group, 21 males and 14 females, 2～13 years old. Interventions T cells subpopulations were determined by indirect immunofluorescence of OKT linesmonoclonal antibodies. The serum IgG was determined by routine simple agar immunodiffusion tests. Results and Conclusions The CD_3~+ and CD_4~+ cells are of no change in the children patients withsimple nephrotic syndrome, and the CD_8~+ and CD_(10)~+ cells are obviously increased, the Values of CD_4~+/CD_8~+ are obviously lower than those in the control qroup, there are no difference between the incipientand relapse groups. The levels of serum IgG were decreased in the 85.3% children patients, IgM were inc-reased in 29.4% of that. The values of CD_4~+/CD_8~+ have positive correlation and negative correlationwith the levels of serum IgG and IgM respectively.