LncRNA GMDS-AS1 inhibits lung adenocarcinoma development by regulating miR-96-5p/CYLD signaling.

According to the global cancer statistic, lung cancer is one of the most dangerous tumors, which poses a serious threat to human health. Exploration the mechanism of lung cancer and new targeted therapeutic measures is always the hot topic. Long noncoding RNA (lncRNA) is an important factor affecting the development of tumors. However, the research on the mechanism of lncRNA in the progress of lung cancer needs to be further expanded. In this study, we found that the expression of lncRNA GMDS-AS1 was significantly reduced in lung adenocarcinoma (LUAD) tissues and cells. Upregulated GMDS-AS1 can significantly inhibit the proliferation of LUAD cells and promote cell apoptosis in vitro and in vivo. The results indicate that GMDS-AS1 acts as a tumor suppressor gene to affect the development of LUAD. Further studies revealed that GMDS-AS1 is a target gene of miR-96-5p, and GMDS-AS1 regulates proliferation and apoptosis of LUAD cells in association with miR-96-5p. In addition, we also confirmed that CYLD lysine 63 deubiquitinase (CYLD) is also a target gene of miR-96-5p. Through various validations, we confirmed that GMDS-AS1 can act as a ceRNA to upregulate the expression of CYLD by sponging miR-96-5p. Moreover, the intervention of GMDS-AS1/miR-96-5p/CYLD network can regulate the proliferation and apoptosis of LUAD cells. In this study, we revealed that the GMDS-AS1/miR-96-5p/CYLD network based on ceRNA mechanism plays an important role in the development of LUAD and provides a new direction and theoretical basis for targeted therapy of LUAD.

18F-fluorodeoxyglucose positron emission tomography/computed tomography in the diagnosis of benign pulmonary lesions in sarcoidosis.

BACKGROUND: Many benign pulmonary lesions, especially sarcoidosis, are metabolically active and are indistinguishable from lung cancer using 18F-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) imaging. This study sought to analyze the 18F-FDG PET/CT imaging features of benign pulmonary lesions and to improve the differential diagnosis of benign pulmonary lesions by 18F-FDG PET/CT imaging. METHODS: One hundred and thirteen patients with benign pulmonary lesions were studied retrospectively. Each patient underwent an 18F-FDG PET/CT scan. All cases were identified by pathology, diagnostic therapy or follow-up. The maximum standardized uptake value (SUVmax) was calculated for each pulmonary lesion. RESULTS: According to the final results, the benign pulmonary lesions were classified as inflammatory lesions (n=77) and granulomas (n=36) by histopathological diagnoses. The SUVmax of inflammatory lesions and granulomas were both high (4.55±2.77 and 6.81±3.96, respectively; P<0.05). When the benign pulmonary lesions were classified by clinical diagnoses, the SUVmax of sarcoidosis was significantly different from other diseases (15.12±5.67; P<0.01). CONCLUSIONS: Inflammatory lesions and granulomas show moderate or high FDG uptake on 18F-FDG PET/CT, but granulomas have higher values. 18F-FDG PET/CT appeared to have a higher SUVmax for the differential diagnosis of sarcoidosis and benign pulmonary lesions.

Toll-like receptor 2 and dectin-1 function as promising biomarker for Aspergillus fumigatus infection.

In recent years, along with the wide application of organ transplantation and immunosuppressive agents, as well as the abuse of broad spectrum antibiotics, the incidence of invasive fungal infections has been increasing gradually. The present study aimed to identify novel biomarkers in cells infected with Aspergillus fumigatus. Human umbilical vein endothelial cells (HUVECs) were infected with Aspergillus fumigatus and then harvested at different time-points (0, 1, 2, 4 and 6 h). The expression Toll-like receptor 2 (TLR2) and dectin-1 expression were examined using flow cytometry and western blotting, and fluorescence-based microscopy was used to evaluate their distribution. The results indicated that TLR2 and dectin-1 protein levels were localized on the surface of HUVECs, and that dectin-1 was distributed on HUVEC membranes as observed under confocal microscope. Immunofluorescence assay result revealed that the optical intensity of dectin-1 in the Aspergillus fumigatus-infected group was significantly increased at 0, 1 and 2 h compared with the control group (P<0.05). However, the optical intensity of TLR2 in the Aspergillus fumigatus-infected group was markedly decreased between 0 and 6 h, as compared with the control group (P<0.05). Western blot analysis indicated that dectin-1 expression was significantly increased and TLR2 expression was significantly decreased at 0, 1 and 2 h post infection in the Aspergillus fumigatus-infected group compared with the control group. Furthermore, the expression of TLR2 was also negatively correlated with the concentration of Aspergillus fumigatus. In conclusion, upon infection of cells with Aspergillus fumigatus, TLR2 and dectin-1 expression levels were significantly altered. Therefore, TLR2 and dectin-1 levels may function as promising biomarkers for the treatment or diagnosis of Aspergillus fumigatus infection.

sRAGE alleviates neutrophilic asthma by blocking HMGB1/RAGE signalling in airway dendritic cells.

Receptor for advanced glycation end products (RAGE) plays a role in inflammatory reactions. The soluble form of RAGE (sRAGE) acts as a decoy to inhibit interactions of RAGE with advanced glycation end products such as High mobility group box 1 (HMGB1). We have demonstrated that HMGB1 directs Th17 skewing by regulating dendritic cell (DC) functions in a previous study. However, the protective effects of HMGB1 blockade with sRAGE in the development of neutrophilic asthma remain unclear. Here, we showed that allergen challenge decreased expression of sRAGE in a murine model of neutrophilic asthma, correlating well with neutrophil counts and interleukin (IL)-17 production. When HMGB1 signalling was blocked by intratracheal administration of sRAGE before sensitisation, HMGB1 expression, neutrophilic inflammation, and Th17-type responses were reduced significantly. Anti-asthma effects of sRAGE were achieved by inhibition of RAGE and IL-23 expression in airway CD11c+ antigen-presenting cells. Finally, we showed that sRAGE inhibited Th17 polarisation induced by recombinant HMGB1 (rHMGB1)-activated dendritic cells (DCs) in vitro. Adoptive transfer of rHMGB1-activated DCs was sufficient to restore airway inflammation, whereas transfer of rHMGB1 plus sRAGE-activated DCs significantly reduced neutrophilic inflammation. Thus, sRAGE prevents Th17-mediated airway inflammation in neutrophilic asthma at least partly by blocking HMGB1/RAGE signalling in DCs.

Adenosine Triphosphate Promotes Allergen-Induced Airway Inflammation and Th17 Cell Polarization in Neutrophilic Asthma.

Adenosine triphosphate (ATP) is a key mediator to alert the immune dysfunction by acting on P2 receptors. Here, we found that allergen challenge caused an increase of ATP secretion in a murine model of neutrophilic asthma, which correlated well with neutrophil counts and interleukin-17 production. When ATP signaling was blocked by intratracheal administration of the ATP receptor antagonist suramin before challenge, neutrophilic airway inflammation, airway hyperresponsiveness, and Th17-type responses were reduced significantly. Also, neutrophilic inflammation was abrogated when airway ATP levels were locally neutralized using apyrase. Furthermore, ATP promoted the Th17 polarization of splenic CD4+ T cells from DO11.10 mice in vitro. In addition, ovalbumin (OVA) challenge induced neutrophilic inflammation and Th17 polarization in DO11.10 mice, whereas administration of suramin before challenge alleviated these parameters. Thus, ATP may serve as a marker of neutrophilic asthma, and local blockade of ATP signaling might provide an alternative method to prevent Th17-mediated airway inflammation in neutrophilic asthma.

Anti-HMGB1 neutralizing antibody ameliorates neutrophilic airway inflammation by suppressing dendritic cell-mediated Th17 polarization.

We demonstrate that high mobility group box 1 protein (HMGB1) directs Th17 skewing by regulating dendritic cell (DC) function. First, our in vitro studies reveal that recombinant HMGB1 (rHMGB1) activates myeloid DCs to produce IL-23 in vitro, and rHMGB1-activated DCs prime naïve lymphocytes to produce the Th17 cytokine IL-17A. Second, we demonstrate that anti-HMGB1 neutralizing antibody attenuates HMGB1 expression, neutrophilic inflammation, airway hyperresponsiveness, and Th17-related cytokine secretion in vivo by using a murine model of neutrophilic asthma induced by ovalbumin (OVA) plus lipopolysaccharide (LPS). Furthermore, anti-HMGB1 neutralizing antibody decreases the number of Th17 cells in lung cells and suppresses the production of IL-23 by lung CD11C(+) APCs. Finally, we show that intranasal adoptive transfer of rHMGB1-activated DCs was sufficient to restore lung neutrophilic inflammation and the Th17 response in a DC-driven model of asthma, whereas the transfer of rHMGB1 plus anti-HMGB1-treated mDCs significantly reduced these inflammation phenotypes. These data suggest, for the first time, that HMGB1 drives the DC-polarized Th17-type response in allergic lung inflammation and that blocking HMGB1 may benefit the attenuation of neutrophilic airway inflammation in asthma.

[The expressions of nerve growth factor and its tyrosine kinase A receptor on alveolar macrophages in a rat model of chronic obstructive pulmonary disease].

OBJECTIVE: To observe the expressions of nerve growth factor (NGF) and its tyrosine kinase A (TrkA) receptor on alveolar macrophage in a rat model of chronic obstructive pulmonary disease (COPD). METHODS: Forty healthy male SD rats were randomly divided into a control group and a COPD group. The COPD model was established by exposing the rats to cigarette smoke for 6 months, and lung function changes were measured. Lung histopathological changes were detected by HE staining. The expression of NGF protein in the supernatant of alveolar macrophage (AM) culture medium was detected by ELISA. Confocal microscopy was used to identify the separation and purification of AM from bronchoalveolar lavage fluid, and to detect semi-quantitatively the expression of TrkA receptor on AM. NGF and its TrkA receptor at the mRNA level were evaluated by real-time PCR. The differences among groups were calculated by one way ANOVA, and comparison between groups was made by t test. RESULTS: Significant decrease of pulmonary compliance [(0.15 ± 0.03) ml/cm H(2)O (1 cm H2O = 0.098 kPa)] and minute ventilation [(0.045 ± 0.004) L], and significant increase of airway resistance [(0.038 ± 0.004) cm H2O×L(-1)×s(-1)] were found in the COPD group compared with the control group [(0.42 ± 0.05) ml/cm H2O and (0.102 ± 0.010) L and (0.016 ± 0.002) cm H2O×L(-1)×s(-1), t = 9.46 - 12.99, respectively, all P < 0.01]. Alveolar wall thinning, alveolar septa breakdown, alveolar enlargement and emphysema were significant in the COPD rats. The expression of NGF protein in the supernatant of AM culture medium was enhanced in the COPD group [(3.79 ± 1.52) ng/L] compared with the controls [(0.94 ± 0.27) ng/L, t = 4.13, P < 0.05]. Mean fluorescence intensity of TrkA protein on AM in the COPD group (19.5 ± 1.5) was higher than that in the control group (11.2 ± 1.9, t = 7.95, P < 0.05). The expressions of NGF and TrkA at mRNA level in the COPD group (24.8 ± 6.0 and 9.0 ± 3.3) were increased compared with the control group (1.0 ± 0.2 and 1.0 ± 0.4, t = 8.48 and 5.16, all P < 0.05). CONCLUSIONS: The expressions of NGF and its TrkA receptor on AM in COPD group were increased, indicating that NGF and its TrkA receptor might be involved in the pathogenesis of COPD mediated by AM.

[Metalloproteinase-9/tissue inhibitor of metalloproteinase-1 in induced sputum in patients with asthma and chronic obstructive pulmonary disease and their relationship to airway inflammation and airflow limitation].

OBJECTIVE: To investigate the levels of metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in induced sputum in patients with asthma and chronic obstructive pulmonary disease (COPD), and its relationship to the number of inflammation cells and lung function. METHODS: Fourteen patients with asthma in remission stages, 12 patients with stable COPD and 10 normal control subjects were included in this study. Lung function was measured. Induced sputum was obtained and processed for cell differential and the supernatant was assayed for the concentrations of interleukin-4 (IL-4), MMP-9 and TIMP-1 by enzyme-linked immunosorbent assay (ELISA). RESULTS: The percentage of eosinophils in induced sputum in asthmatics (0.181 +/- 0.067) was significantly higher than that in normal control subjects (0.007 +/- 0.005) and in COPD (0.042 +/- 0.017, F = 4.32, P < 0.05). The percentage of neutrophils in induced sputum in patients with COPD (0.500 +/- 0.101) was significantly higher than that in asthmatics (0.30 +/- 0.07) and in normal control subjects (0.26 +/- 0.06, F = 4.13, P < 0.05). The concentrations of IL-4 in asthmatics, COPD and normal control subjects [respectively, (19 +/- 7) x 10(-3) g/L, (14 +/- 6) x 10(-3) g/L, (11 +/- 4) x 10(-3) g/L] did not show significant difference (F = 1.56, all P > 0.05) and did not correlate with the number of eosinophils (r = 0.33, 0.11, 0.19, all P > 0.05) and neutrophil (r = 0.25, 0.39, 0.40, all P > 0.05) and FEV(1) values (predicted r = 0.21, 0.35, 0.17, all P > 0.05). The concentrations of MMP-9 and TIMP-1 in induced sputum in asthmatics [(15.9 +/- 6.0) g/L, (19.8 +/- 8.5) g/L, respectively] and COPD [(13.4 +/- 5.1) g/L, (16.7 +/- 7.6) g/L, respectively] were significantly higher than those in normal control subjects [(1.8 +/- 1.1) g/L, (1.3 +/- 0.9) g/L, respectively] (F = 2.99, 4.22, respectively, all P < 0.05). Increased concentration of MMP-9 correlated positively with the percentage of eosinophils in asthmatics (r = 0.71, P < 0.05) and with the percentage of neutrophils in COPD (r = 0.59, P < 0.05), but did not correlate with FEV(1) values (predicted r = 0.22, 0.16, all P > 0.05) and FEV(1)/FVC (r = 0.25, 0.30, all P > 0.05). Increased concentration of TIMP-1 did not correlate with the number of eosinophils (r = 0.27, 0.31, all P > 0.05) and neutrophil (r = 0.20, 0.35, all P > 0.05) in asthmatics and COPD, but correlated inversely with FEV(1) values (predicted, respectively, r = -0.58, -0.62, all P < 0.05). The ratio of MMP-9/TIMP-1 was significantly lower in asthmatics 0.8 +/- 0.7 and COPD 0.8 +/- 0.6 than that in normal control subjects (1.5 +/- 0.6, F = 3.70, P < 0.05). The ratio was not statistically different between asthmatics and COPD (F = 1.78, P > 0.05). In asthmatics and COPD patients, the ratio of MMP-9/TIMP-1 in induced sputum correlated positively with FEV(1%) (respectively, r = 0.56, 0.61, all P < 0.05). CONCLUSION: An imbalance between MMP-9 and TIMP-1 in induced sputum in asthmatics and COPD is associated with airway inflammation and airflow limitation, which may play a role in the pathogenesis of extracellular matrix remodeling and airflow limitation.