LncRNA-MEG3 functions as a competing endogenous RNA to regulate Treg/Th17 balance in patients with asthma by targeting microRNA-17/ RORγt.

BACKGROUND: Treg/Th17 imbalance plays an essential role in the pathogenesis of asthma. Disordered LncRNAs were observed in asthma, however, whether LncRNAs can regulate the Treg/Th17 balance and its mechanism still needs to be investigated. METHODS: Microarrays were performed to identify the differentially expressed lncRNAs and microRNAs in peripheral blood CD4 + T cells from patients with asthma and healthy controls. Bioinformatical evidence was used to select candidate lncRNAs and microRNAs which may involve in regulation of Treg/Th17 balance. The function of LncRNA-MEG3 and microRNA-17 on the alteration of the CD4 + T cell population were determined in vitro experiments. Meanwhile, the regulatory effect of LncRNA-MEG3 and microRNA-17 on RORγt or Foxp3 was estimated. The interaction of LncRNA-MEG3 with microRNA-17 was confirmed by dual luciferase reporter assay and RNA pull-down. RESULTS: 25 lncRNAs and 19 microRNAs were selected as candidate genes which differentially expressed in CD4 + T cells from patients with asthma compared with healthy controls and had potential to control Treg/Th17 balance by regulating RORγt or Foxp3. Alternation of LncRNA-MEG3 changed the function and increased the percentage of Th17. LncRNA-MEG3 could regulate the RORγt mRNA and protein level. LncRNA-MEG3 could inhibit the level of microRNA-17 as a competing endogenous RNA (ceRNA). microRNA-17 suppressed Th17 though targeting RORγt directly. CONCLUSION: LncRNA-MEG3 can sponge microRNA-17 as a ceRNA, thereby regulating RORγt and ultimately affecting Treg/Th17 balance in asthma. The lncRNA/microRNA axis may have potential application in clinical treatment and diagnosis of the disease.

Clinical features of secondary pulmonary alveolar proteinosis associated with myelodysplastic syndrome: Two case reports.

RATIONALE: Pulmonary alveolar proteinosis (PAP) is a rare lung disorder characterized by the abnormal accumulation of alveolar surfactant protein in alveolar spaces. Secondary PAP can result from myelodysplastic syndrome (MDS). PATIENT CONCERNS: But most reports described a single case; here we reported 2 cases of PAP secondary to MDS. One case developed secondary PAP at the same time as MDS, and the other developed during the course of MDS. DIAGNOSES: The diagnosis of PAP was made by bronchoalveolar lavage and based on the identification of periodic acid-Schiff-positive proteinaceous material. Chest high resolution CT (HRCT) scans showed variable distribution of ground glass opacities, but crazy-paving appearance was not seen in our 2 cases. INTERVENTIONS: Because the patients' general conditions were poor, whole lung lavage was not used in the 2 cases. OUTCOMES: And the 2 cases' prognoses were poor. LESSONS: In conclusion, pulmonary physicians should suspect the possibility of secondary PAP when they encounter unexplained pulmonary infiltrates with some hematologic or infectious disease that shows diffuse bilateral GGO on an HRCT scan.

miR-371, miR-138, miR-544, miR-145, and miR-214 could modulate Th1/Th2 balance in asthma through the combinatorial regulation of Runx3.

Asthma is tightly related to the imbalance of Th1/Th2 cells, and Runx3 plays a pivotal role in the differentiation of T helper cells. The present study aimed to investigate dysregulated microRNAs that may target Runx3 in CD4+ T cells from asthmatic patients and reveal Runx3 function in Th1/Th2 balance regulation. We detected the levels of Th1- and Th2-related cytokines by ELISA and analyzed the differentiation marker gene of T helper cells by qRT-PCR. Results indicated that an imbalance of Th1/Th2 cells was present in our asthmatic subject. Runx3 expression was reduced in the CD4+ T cells from asthmatic patients. Overexpression of Runx3 could restore the Th1/Th2 balance. After performing microRNA microarray assay, we found a series of microRNAs that were considerably altered in the CD4+ T cells from asthmatic patients. Among these upregulated microRNAs, eight microRNAs that may target Runx3 were selected by bioinformatics prediction. Five microRNAs, namely miR-371, miR-138, miR-544, miR-145, and miR-214, were confirmed by qRT-PCR and selected as candidate microRNAs. Luciferase reporter assay showed that these five microRNAs could directly target the 3'-UTR of Runx3. However, only simultaneous inhibition of these five microRNAs could alter the expression of Runx3. Most importantly, only simultaneous inhibition could improve the Th1/Th2 balance. Thus, we suggest that miR-371, miR-138, miR-544, miR-145, and miR-214 can modulate the Th1/Th2 balance in asthma by regulating Runx3 in a combinatorial manner.

1,25-dihydroxyvitamin D3 reduces mouse airway inflammation of neutrophilic asthma by transcriptional modulation of interleukin-17A.

Corticosteroid resistance and severe airflow obstruction have been proved to participate in the neutrophilic inflammation of airway in uncontrollable asthmatics. IL-17 is one of the pro-inflammatory cytokines produced by Th17 cells, and it plays an important role in the neutrophilic inflammation of airway in steroid-resistant asthmatics. Recent data have proved that 1,25(OH)2D3 represses IL-17A in inflammation and Th17-mediated autoimmunity through vitamin D receptors(VDR) at the level of transcription. Our study validated that 1,25-(OH)2D3 can modulate IL-17A on the transcriptional level by using Runx1, thus reducing inflammation in the airway of mice with neutrophilic asthma. 1,25(OH)2D3 may be promising for the therapeutic applications of neutrophilic asthma.

Protective effects of astragaloside IV against ovalbumin-induced lung inflammation are regulated/mediated by T-bet/GATA-3.

Bronchial asthma is characterized by chronic lung inflammation, airway hyperresponsiveness, and airway remodelling. Astragaloside IV (3-O-ß-D-xylopyranosyl-6-O-ß-D-glucopyranosyl-cycloastragenol, AST), the primary pure saponin isolated from the root of Astragalus membranaceus, is an effective compound with distinct pharmacological effects including anti-inflammation, immunoregulation, and antifibrosis. However, the effect of AST on asthma remains unclear. In the present study, in the murine model of asthma, the airway hyperresponsiveness was relieved after treatment with AST, accompanied by a reduction of inflammatory cells. In addition, the levels of IL-4 and IL-5 decreased, while the IFN-γ level increased, in bronchoalveolar lavage fluid. The compound also significantly inhibited the synthesis of GATA-3-encoding mRNA and protein in addition to increasing the synthesis of T-bet-encoding mRNA and protein in both lung tissues and CD4+ T cells. Our findings indicate that AST treatment inhibits ovalbumin-induced airway inflammation by modulating the key master switches GATA-3 and T-bet, which results in committing T helper cells to a Th1 phenotype.

[The clinicopathological features of acute fibrinous and organizing pneumonia].

OBJECTIVE: To improve understanding of the clinical, radiological and pathological characteristics of acute fibrinous and organizing pneumonia (AFOP). METHODS: The clinical data of 5 AFOP patients were retrospectively analyzed. AFOP was diagnosed via percutaneous lung biopsy guided by chest computerized tomography (CT) in the Affiliated Drum Tower Hospital of Nanjing University Medical School during March 2011 to June 2012. The clinical, radiological and pathological characteristics of those patients were summarized. RESULTS: Among the 5 patients, 2 were male and 3 were female, aging 43-61 years. They were all subacute onset. The main clinical manifestations were dyspnea, productive cough, fever and chest pain with hypoxemia via blood gas analysis. Bilateral infiltrates with diffuse and pathy distribution were the predominant features in chest HRCT. The pathological examination revealed slightly widened alveolar septa, 1ymphocyte and plasma cell infiltration and the presence of intra-alveolar fibrin in the form of fibrin "balls" (organization) within the alveolar spaces. No neutrophil, and eosinophil infiltration and hyaline membrane formation were detected, which was different from other well-recognized histologic patterns of acute lung injury, such as diffuse alveolar damage, cryptogenic organizing pneumonia and eosinophilic pneumonia. All patients were treated by corticosteroids and showed significant clinical and radiological improvement. CONCLUSIONS: AFOP has nospecific features, and its diagnosis depends on pathological examination. Treatment with corticosteroids is optimal. However, whether it is a unique interstitial disease needs to be further clinically investigated.

Beta(2)-adrenergic receptor haplotype/polymorphisms and asthma susceptibility and clinical phenotype in a Chinese Han population.

Association and linkage studies of beta2-adrenergic receptor (beta2AR) polymorphisms in relation to the expression of asthmatic phenotypes and immune regulatory mechanisms have shown inconsistent results. This study was designed to analyze the relationship of particular combinations of single nucleotide polymorphisms (SNPs) or haplotypes of the beta2AR gene with bronchial asthma, bronchodilator response, and total IgE. By direct DNA sequencing, five SNPs (in positions -47, -20, 46, 79, and 252) of beta2AR gene were determined and combined with haplotypes in 201 asthmatic patients and 276 normal controls recruited from the Chinese Han population. Significantly higher bronchodilator response was observed in patients with homozygotic genotype 46A/A (13.40 ± 3.48%), compared with those with homo-46G/G (7.25 ± 3.11%) and heterozygotes 46A/G (7.39 ± 3.14%), respectively (p < 0.0001). There was also a significant difference in bronchodilator response when beta2AR haplotypes were analyzed (p = 0.003). From two common SNPs at positions 46A/G and 79C/G, we had determined three haplotypes that constructed six haplotype pairs. Comparison of the mean delta forced expiratory volume in 1 second (FEV1) values for the six haplotype pairs showed significant difference. Subjects homozygous for 46A/79C (Arg16/Gln27) had the highest deltaFEV1 (13.40 ± 3.48%) and those with 46G/79C (Gly16/Gln27) homozygote had the lowest (6.43 ± 0.55%). The two SNP haplotype pairs were significantly associated with delta FEV1 (p < 0.0001). Significantly higher total IgE levels were found in patients with homozygotic carriers of 79C genotypes (p = 0.022) and homozygotic haplotype -47 T/-20 T/46 A/79 C/252 G (p < 0.0001). These results indicate that the manifestation of asthma might be affected by either an individual beta2AR SNPs or beta2AR haplotype.

[Association between beta2-adrenergic receptor haplotype/polymorphisms and asthma phenotype].

OBJECTIVE: To analyze the association of single nucleotide polymorphisms (SNP)/ haplotypes of beta2-adrenergic receptor (beta2-AR) gene with bronchodilator response and total serum immunoglobulin E (IgE). METHODS: Five SNP (in positions: -47, -20, 46, 79, 252) genotypes of beta2-AR gene in 201 asthmatic patients and 276 controls were determined by direct DNA sequencing, and the haplotypes were combined from 2006 February to 2007 February. Statistical analyses were performed with SPSS 11.5 software. The genotype frequencies for each polymorphism were tested for deviation from the Hardy-Weinberg equilibrium by chi2 goodness-of-fit analysis. The frequencies of the five polymorphic genotypes were compared with chi2 test, and the degree of linkage disequilibrium occurring between these polymorphic loci were analyzed by fisher' s exact. For the comparison of quantitative traits among different genotypes/haplotypes, ANOVA was used. Least significant difference (LSD) was used if there was statistical significant. RESULTS: The bronchodilator response in patients with Argl6Argl6 was (13 +/- 4)L, significantly higher than those with Arg16Gly16 [(7 +/- 3) L and Gly16Gly16 (7 +/- 3)L, F = 81.55, P < 0.01]. When beta2-AR haplotypes were analyzed, bronchodilator response was highest in patients with haplotype Arg16Gln27/Arg16Gln27 [(13.4 +/- 3.5) L], compared to that with Gly16Gln27/Gly16Gln27 [(6.4 +/- 0.6) L], Gly16Glu27/Gly16Glu27 [(7.6 +/- 3.1)L], Gly16Gln27/Gly16Glu27 [(6.9 +/- 3.5)L], Gly16Gln27/Arg16Gln27 [(7.2 +/- 3.3) L, and Gly16Glu27/Arg16Gln27 (7.9 +/- 2.7) L, F = 32.55, P < 0.01]. The total IgE level was (2.51 +/- 0.33) IU/L in patients with genotype Gln27Gln27, significantly higher than that with Gln27Glu27 [(2.30 +/- 0.82) IU/L, F = 3.89, P < 0.05]. The total IgE level was significantly lowest in patients with haplotype Gly16Gln27/Arg16Gln27 (2.13 +/- 0.15) IU/L, compared to that with Arg16Gln27/Arg16Gln27 (2.56 +/- 0.14) IU/L, Gly16Glu27/Gly16Glu27 (2.40 +/- 0.16) IU/L, Gly16Gln27/Gly16Glu27 (2.54 +/- 1.26) IU/L, Gly16Gln27/Arg16Gln27 [(2.48 +/- 0.48) IU/L, F = 3.56, P < 0.01]. CONCLUSIONS: These results indicate that depending on phenotypes studied, either an individual beta2-AR SNP or haplotype might affect disease manifestations.

Association between beta2-adrenergic receptor genetic polymorphisms and total serum IgE in asthmatic patients of Chinese Han nationality.

BACKGROUND: beta(2)-Adrenergic receptor (beta(2)-AR) polymorphisms occurring at amino acid position 16 (Arg-Gly) and 27 (Gln-Glu) are known to be functionally relevant and also disease-modifying in subjects with asthma. It has been found in Caucasoid asthmatic patients that the Gln27 genotype beta(2)-AR was associated with an increase in total serum IgE levels. The association between beta(2)-AR genetic polymorphisms and total serum IgE in asthmatic patients of Chinese Han nationality remains to be established. OBJECTIVES: It was the aim of this study to investigate the association between beta(2)-AR genetic polymorphisms and total serum IgE in asthmatic patients of Chinese Han nationality. METHODS: All 59 asthmatic patients investigated (27 males and 32 females, aged between 16 and 60 years) were people of Chinese Han nationality. They were tested for their total serum IgE levels with the enzyme-linked immunosorbent assay test, and beta(2)-AR genetic polymorphisms were tested with the polymerase chain reaction allele-specific oligonucleotide hybridization assay. RESULTS: There was a significant difference of serum IgE levels among three beta(2)-AR 27 loci groups (p < 0.0001), with the highest IgE level [(1.24 +/- 0.25) x 10(6) IU/l] in the Gln/Gln group and the lowest IgE level [(0.48 +/- 0.06) x 10(6) IU/l] in the Glu/Glu group. No polymorphism of beta(2)-AR 16 loci was found to be associated with total serum IgE (p > 0.05). CONCLUSIONS: Our research suggested that in asthmatic patients of Chinese Han nationality, the beta(2)-AR genetic polymorphism at 27 loci could be associated with serum IgE levels and it might therefore play an important role in the determination of phenotypes of bronchial asthma.