Hydrogen/oxygen therapy for the treatment of an acute exacerbation of chronic obstructive pulmonary disease: results of a multicenter, randomized, double-blind, parallel-group controlled trial.

BACKGROUND: To investigate whether the administration of hydrogen/oxygen mixture was superior to oxygen in improving symptoms in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). METHODS: This prospective, randomized, double-blind, controlled clinical trial in 10 centres enrolled patient with AECOPD and a Breathlessness, Cough, and Sputum Scale (BCSS) score of at least 6 points. Eligible patients were randomly assigned (in a 1:1 ratio) to receive either hydrogen/oxygen mixture or oxygen therapy. Primary endpoint was the change from baseline in BCSS score at day 7. Adverse events (AEs) were recorded to evaluate safety. RESULTS: Change of BCSS score in Hydrogen/oxygen group was larger than that in Oxygen group (- 5.3 vs. - 2.4 point; difference: - 2.75 [95% CI - 3.27 to - 2.22], meeting criteria for superiority). Similar results were observed in other time points from day 2 through day 6. There was a significant reduction of Cough Assessment Test score in Hydrogen/oxygen group compared to control (- 11.00 vs. - 6.00, p < 0.001). Changes in pulmonary function, arterial blood gas and noninvasive oxygen saturation did not differ significantly between groups as well as other endpoints. AEs were reported in 34 (63.0%) patients in Hydrogen/oxygen group and 42 (77.8%) in Oxygen group. No death and equipment defects were reported during study period. CONCLUSIONS: The trial demonstrated that hydrogen/oxygen therapy is superior to oxygen therapy in patient with AECOPD with acceptable safety and tolerability profile. TRIAL REGISTRATION: Name of the registry: U.S National Library of Medicine Clinical Trials; Trial registration number: NCT04000451; Date of registration: June 27, 2019-Retrospectively registered; URL of trial registry record: https://www.clinicaltrials.gov/ct2/show/study/NCT04000451?term=04000451&draw=2&rank=1 .

LncRNA-DANCR contributes to lung adenocarcinoma progression by sponging miR-496 to modulate mTOR expression.

Long non-coding RNAs (lncRNAs) have emerged as new and important regulators of pathological processes including tumour development. In this study, we demonstrated that differentiation antagonizing non-protein coding RNA (DANCR) was up-regulated in lung adenocarcinoma (ADC) and that the knockdown of DANCR inhibited tumour cell proliferation, migration and invasion and restored cell apoptosis rescued; cotransfection with a miR-496 inhibitor reversed these effects. Luciferase reporter assays showed that miR-496 directly modulated DANCR; additionally, we used RNA-binding protein immunoprecipitation (RIP) and RNA pull-down assays to further confirm that the suppression of DANCR by miR-496 was RISC-dependent. Our study also indicated that mTOR was a target of miR-496 and that DANCR could modulate the expression levels of mTOR by working as a competing endogenous RNA (ceRNA). Furthermore, the knockdown of DANCR reduced tumour volumes in vivo compared with those of the control group. In conclusion, this study showed that DANCR might be an oncogenic lncRNA that regulates mTOR expression through directly binding to miR-496. DANCR may be regarded as a biomarker or therapeutic target for ADC.

[Modulation of TLR9 on anti-tumor immune responses of peripheral blood mononuclear cells from patients with non-small-cell lung cancer].

OBJECTIVE: To assess the modulation of TLR9 on anti-tumor immune responses in peripheral blood mononuclear cells (PBMC) from patients with non-small-cell lung cancer (NSCLC). METHODS: PBMCs were isolated from 36 NSCLC patients. Lung cancer cells were isolated from these patients and further enriched. PBMCs were cultured in RPMI-1640 medium (blank control group), and medium with cytosine guanine oligodeoxynucleotide (CpG ODN, an TLR9 agonist) or control ODN for 72 h; and then flow cytometry was used to examine the expression of CD69 antigen on the surface of CD3 cells, [3H]-thymidine incorporation method was used to examine the cell proliferation, and the IFN-alpha level in the supernatant was measured. Another PBMCs were cultured in medium with interleukin (IL)-1 and then CpG ODN, control ODN, and CpG ODN + chloroquine or inhibitory ODN were added respectively for 24-48 h. Then the IFN-alpha in the supernatant was measured. Subsets were assessed by flow cytometry and the expression of TLR9-mRNA in freshly isolated PBMC was detected by RT-PCR. The production of interferon (IFN)-alpha in the PBMCs was measured by ELISA. The proliferation of the PBMCs was determined by [3H]-thymidine incorporation. The PBMCs co-cultured with CpG ODN and autologous lung tumor cells treated with mitomycin C were used as effector cells, and K562 cells and autologous tumor cells were used as target cells flow cytometry was used to detect the capacity of PBMCs to kill autologous lung tumor cells and K562 cells. Meanwhile we investigated the intracellular expression of IFN-gamma and IL-4 in CD8+ T. RESULTS: The expression level of TLR9 of the PBMCs from patients was not significantly different from that of the PBMCs from the healthy donors. The proportion of CD69 antigen expressing CD3+ T cells of the CpG ODN group was (39.5 +/- 8.9)%, significantly higher than those of the blank control group [(8.8 +/- 1.2)%, t = 40.30, P = 0.00] ands control ODN group [(10.6 +/- 1.0)%, t = 41.85, P = 0.00]. Examination with beta liquid scintillation counter showed that the cpm value of the CpG ODN group was (1.61 +/- 0.20) x 10(4), significantly higher than those of the blank control group [(0.27 +/- 0.14) x 10(4), t = 20.43, P = 0.00] and control ODN group [(0.34 +/- 0.13) x 10(4), t =20.20, P = 0.00]. Chloroquine and inhibitory ODN dose-dependently inhibited the IFN-alpha levels in the supernatant. The CD4 + T/CD8 + T of the CpG ODN group was (3.44 +/- 0.20), significantly higher than those of the control ODN group (1.73 +/- 0.27, t = 19.85, P = 0.00) and blank control group (1.69 +/- 0.13, t = 29.32, P = 0.00). The IFN-gamma positive CD8 + T cells of the CpG ODN group was (18.5 +/- 4.2)%, significantly higher than those of the control ODN group [(4.2 +/- 1.0)%, t = 24.12, P = 0.00] and blank control group [(3.1 +/- 1.2)%, t = 25.1, P = 0.00]. There was no significant differences in the proportion of IL-4 positive CD8 + T cells among different groups. When the E/T was 40:1 the killing capacity of PBMCs against the K562 cells was (19.5 +/- 1.0), significantly higher than those of the control ODN group (7.9 +/- 1.1, t = 19.9, P = 0.00) and blank control group (5.1 +/- 1.6, t = 21.9, P = 0.00), and the killing capacity of PBMCs against the autologous lung tumor cells was (29.8 +/- 2.1), significantly higher than those of the control ODN group (8.1 +/- 0.9, t = 36.9, P = 0.00) and blank control group (5.7 +/- 1.6, t = 35.7, P = 0.00). CONCLUSION: TLR9 signal takes part in the immunomodulation of PBMCs. The activation of TLR9 results in enhanced anti-tumor response in the PBMCs against autologous lung cancer cells and K562 cells.

Continuous tracheal gas insufflation during protective mechanical ventilation in juvenile piglets with acute lung injury induced by endotoxin.

BACKGROUND: Low tidal volume mechanical ventilation is difficult to correct hypoxemia, and prolonged inhalation of pure oxygen can lead to oxygen poisoning. We suggest that continuous tracheal gas insufflation (TGI) during protective mechanical ventilation could improve cardiopulmonary function in acute lung injury. METHODS: Totally 12 healthy juvenile piglets were anesthetized and mechanically ventilated at PEEP of 2 cmH2O with a peak inspiratory pressure of 10 cmH2O. The piglets were challenged with lipopolysaccharide and randomly assigned into two groups (n=6 each group): mechanical ventilation (MV) alone and TGI with continuous airway flow 2 l/min. FIO2 was set at 0.4 to avoid oxygen toxicity and continuously monitored with an oxygen analyzer. RESULTS: Tidal volume, ventilation efficacy index and mean airway resistant pressure were significantly improved in the TGI group (P<0.01 or P<0.05). At 4 hours post ALI, pH decreased to below 7.20 in the MV group, and improved in the TGI group (P<0.01). Similarly, PaCO2 was stable and was significantly lower in the TGI group than in the MV group (P<0.01). PaO2 and PaO2/FIO2 increased also in the TGI group (P<0.05). There was no significant difference in heart rate, respiratory rate, mean artery pressure, central venous pressure, dynamic lung compliance and mean resistance of airway between the two groups. Lung histological examination showed reduced inflammation, reduced intra-alveolar and interstitial patchy hemorrhage, and homogenously expanded lungs in the TGI group. CONCLUSION: Continuous TGI during MV can significantly improve gas exchange and ventilation efficacy and may provide a better treatment for acute lung injury.

Tracheal gas insufflation with partial liquid ventilation to treat LPS-induced acute lung injury in juvenile piglets.

OBJECTIVES: Partial liquid ventilation (PLV) with perfluorocarbons (PFC) seems not superior to conventional ventilation clinically. We hypothesized that a combination of continuous tracheal gas insufflation (TGI) with protective strategy of PLV (low dose of PFC, low inflation pressure, moderate inhalation of oxygen and moderate anesthesia) would improve cardiopulmonary function in acute lung injury. METHODS: Twenty-four healthy juvenile piglets were anesthetized and mechanically ventilated at PEEP of 2 cmH(2)O with a peak inspiratory pressure of 10 cmH(2)O and FIO(2) of 0.4. The piglets were challenged with lipopolysaccharide and randomly assigned to four groups (n = 6 each): (1) mechanical ventilation alone (MV); (2) PLV with perfluorodecalin (10 ml/kg); (3) TGI with continuous airway flow 2 L/min; and (4) combination of PLV and TGI. The outcome was assessed functionally and histologically. RESULTS: All treatments except MV improved pH, PaO(2)/FIO(2), PaCO(2), ventilation efficacy index (VEI) and tidal volume. Both PLV-associated treatments also improved heart rate, respiratory rate, pulse contour cardiac output, systemic vascular resistance, dynamic lung compliance, mean airway resistance and mean airway pressure. The combination group resulted in higher PaO(2)/FIO(2), VEI and a better lung histology score than any other treatments. CONCLUSIONS: The new protective strategy may provide a better treatment for sepsis-induced acute lung injury.

Partial liquid ventilation confers protection against acute lung injury induced by endotoxin in juvenile piglets.

To investigate the effect of partial liquid ventilation (PLV) at low inflation pressures on acute lung injury (ALI), endotoxin was administered to healthy anesthetized juvenile piglets. The animals were randomly assigned to two groups, n=6 each: (1) conventional mechanical ventilation (MV) and (2) PLV with perfluorodecalin (10 mL kg(-1)). Compared with MV, PLV improved each cardiopulmonary variable measured. These variables included pulse contour cardiac output, heart rate, blood pH, breathing rate, both partial pressure of arterial oxygen (PaO2) and PaO2/FIO2 (fraction of inspired oxygen), partial pressure of arterial carbon dioxide (PaCO2), dynamic lung compliance, tidal volume, and ventilation efficacy index. Lung morphology also showed less damage in the PLV group, even in non-dependent regions (P<0.05). Our data support the hypothesis that PLV can decrease pulmonary damage, improve gas exchange and cardiac output, and may lead to a better prognosis in endotoxin-induced ALI.

Functional expression of TLR9 is associated to the metastatic potential of human lung cancer cell: functional active role of TLR9 on tumor metastasis.

CpG-oligonucleotides (CpG-ODN), which induce signaling through Toll-like receptor 9 (TLR9), are widely used as adjuvants in therapy against cancer. However, tumor cells express functional TLR9 were recently reported and the immune effect of CpG ODN on tumor cells remains unclear. Here we investigated the direct effects of CpG ODN on human tumor cell line 95D cells using flow cytometric analysis and Western blotting. We found strongly high expression of TLR9 in 95D cells. Stimulation of 95D cells with CpG ODN induced significantly elevated secretion of IL-1alpha and IL-8, as well as the expression of CXCR4, ICAM-1 and MMP-2. Furthermore, the invasion of 95D cells and TLR9 modifying 95C cells were significantly enhanced by stimulation of CpG ODN, which could be abrogated by inhibitory CpG ODN and chloroquine. These results suggest that functionally active TLR9 is expressed on human tumor cell lines, and may represent a novel insight on the role of TIL9 agonist used in tumor immunotherapy.