Objective To investigate the role of Toll?like receptor 4(TLR4)in acute lung injury induced by paraquat(PQ)poisoning. Meth?ods Twenty wild?type mice(C57BL/6J)and TLR4 knockout mice(C57BL/10ScN)were randomly divided into 4 groups:WT group,WT+PQ group,TLR4 gene knockout(TLR4?ko)group and TLR4?ko+PQ group,with 10 mice in each group. After paraquat treatment for 24 hours,mice were euthanized and sacrificed. TLR4 expression and pro?inflammatory cytokines were evaluated. Results After 24 hours of intraperitoneal injec?tion of PQ,wild?type mice showed significant acute lung injury,including decreased oxygen tension in blood gas,pathological changes in the acute stage of lung injury and increased TLR4 expression(accompanied by increased levels of TNF?α,IL?1βand NF). TLR4?deficient mice exhibited significantly reduced level of paraquat?induced lung injury. Conclusion TLR4 may be required as a mediator and play an important role in acute lung injury induced by paraquat. Inhibition of TLR4 signaling has protection of lung injury induced by PQ.

ObjectiveTo investigate the effect of hydrogen (H2 ) on acute lung injury (ALI) in septic mice.MethodsOne hundred and twelve male C57BL/6 mice,aged 5 weeks,weighing 20-25 g,were randomly divided into 4 groups ( n =28 each):sham operation group (group A),sham operation + H2 group (group B),sepsis group (group C) and sepsis + H2 group (group D).Sepsis was produced by cecum ligation and puncture (CLP).Groups B and D received 1 h inhalation of 2％ H2 at 1 and 6 h after CLP operation or sham operation.Twenty animals in each group were selected and observed for the 7 d survival rate.The left 8 animals in each group were sacrificed at 24 h after CLP operation.Venous blood samples and lung tissues were obtained to determine the levels of superoxide dismutase (SOD),catalase (CAT) and 8-iso-prostaglandin F2alpha (8-iso-PGF2α) in the serum and lungs,the concentration of protein in bronchoalveolar lavage fluid (BALF),and the activity of myeloperoxidase (MPO) in the lungs.The lung injury score (LIS) was assessed and W/D lung weight ratio was calculated.ResultsCompared with group A,the 7 d survival rate and activities of SOD and CAT in the serum and lungs were significantly decreased,and LIS,W/D ratio,the concentration of protein in BALF,MPO activity and 8-iso-PGF2α level in the serum and lungs were significantly increased in group C ( P ＜ 0.05 ).Compared with group C,the 7 d survival rate and activities of SOD and CAT in the serum and lungs were significantly increased,and LIS,W/D ratio,the concentration of protein in BALF,MPO activity and 8-iso-PGF2α level in the serum and lungs were significantly decreased in group C ( P ＜ 0.05).ConclusionH2 can alleviate ALI in septic mice via inhibiting oxidative stress response.

Objective To investigate the effects of hydrogen-rich saline on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice.Methods Thirty-two male C57BL/6 mice,weighing 20-25 g,were randomly divided into 4 groups (n =8 each):control group (group C),hydrogen-rich saline group (group H2),ALI group and ALI + hydrogen-rich saline group (group ALI+ H2).ALI was induced by inhaled aerosolized LPS 25μg (dissolved in PBS) in groups ALI and ALI+ H2.Aerosolized PBS 50μl was inhaled in groups C and H2.0.6mmol/L hydrogen-rich saline 5 ml/kg was injected intraperitoneally at 1 and 12 h after aerosolized PBS or LPS was inhaled in groups H2 and ALI+ H2,respectively.At 24 h after PBS or LPS treatment,the animals were mechanically ventilation for 15 min,and arterial blood gas analysis was performed.The oxygenation index (PaO2/FiO2) was calculated.Bronchoalveolar lavage fluid (BALF) was collected for determination of total protein concentration and polymorphonuclear neutrophils (PMN) count.The concentrations of TNF-α,IL-1β,IL-6 and high mobility group box-1 (HMGB-1) protein in BALF were determined by ELISA.Then the animals were sacrificed and the lungs were obtained for determination of W/D lung weight ratio (W/D ratio),and myeloperoxidase (MPO) and caspase-3 activities.The pathological changes of the lung were scored.Apoptosis index (AI) was calculated.Results Compared with group C,no significant difference was found in PaO2/FiO2,the concentrations of protein,TNF-α,IL-1β,IL-6 and HMGB1,and PMN count in BALF,lung injury score,W/D ratio,MPO and caspase-3 activities and AI in group H2 (P ＞ 0.05),and PaO2/FiO2 was significantly decreased,the concentrations of total protein,TNF-α,IL-1β,IL-6 and HMGB1,and PMN count in BALF,lung injury score,W/D ratio,MPO and caspase-3 activities,and Al were increased in groups ALI and ALI + H2 (P ＜ 0.05).Compared with group ALI,PaO2/FiO2 was signifieantly increased,the concentrations of total protein,TNF-α,IL-1β,IL-6 and HMGB1,and PMN count in BALF,lung injury score,W/D ratio,MPO and caspase-3 activities,and AI were decreased in group ALI+ H2 (P ＜ 0.05).Conclusion Hydrogen-rich saline can alleviate LPS-induced ALI via inhibition of inflammatory response and apoptosis in mice.

Objective To investigate the effect of inhalation of hydrogen (H2) on the inflammation response during sepsis-induced acute lung injury in mice.Methods Forty-eight male ICR mice,aged 5 weeks,weighing 20-25 g,were randomly divided into 4 groups (n =12 each):sham operation group (group A),sham operation + H2 group (group B),sepsis group (group C) and sepsis + H2 group (group D).Sepsis was produced by cecal ligation and puncture (CLP).Groups B and D received 1 h inhalation of 2 ％ H2 at 1 and 6 h after CLP or sham operation.The behavior was assessed and scored at 24 h after CLP.Arterial blood samples were taken for blood gas analysis.Oxygenation index was calculated.Venous blood samples were taken for determination of the levels of serum tumor necrosis factor-α (TNF-α),interleukin-1 (IL-1β),high mobility group protein B1 (HMGB1) and interleukin-10 (IL-10).The mice were then sacrificed and lungs were removed for determination of the levels of TNF-α,IL-1β,HMGB1 and IL-10 in lung tissues.The apoptotic index was calculated.Results Compared with group A,the behavior score,apoptotic index and levels of TNF-α,IL-1β,HMGB1 and IL-10 were increased,and the oxygenation index was decreased in groups C and D,and no significant change was found in the parameters mentioned above in group B.Compare with group C,the oxygenation index and level of IL-10 were significantly increased,and the behavior score,apoptotic index and levels of TNF-α,IL-1β and HMGB1 were significantly decreased in group D.Conclusion Inhalation of H2 can regulate the balance between pro-and anti-inflammatory cytokines,inhibit the inflammation response and alleviate sepsis-induced acute lung injury in mice.

Objective To investigate the effect of hydrogen gas on lipopolysaccharide (LPS)-induced apoptosis in human umbilical vein endothelial cells (HUVECs) in vitro.Methods HUVEC-12 cells were seeded in 96-well plates with a density of 1 × 104/ml (200 μl/hole) or in 6-well plates (2 ml/hole) with a density of 1 × 106/ml and randomly divided into 4 groups (n =30 each):control group (group C),hydrogen gas (H2) group,LPS group and LPS + H2 group.The cells were cultured in the plain culture medium in groups C and LPS or in hydrogen-saturated culture medium in groups H2 and LPS + H2.In addition,LPS 1 μg/ml was added simultaneously in groups LPS and LPS + H2 and the equal volume of normal saline was added instead in groups C and H2.The cell viability and apoptosis were measured by MTT assay and flow cytometry respectively after 24 h incubation.The concentration of high-mobility group box 1 (HMGB1) in the supernatant was determined by ELISA.Results Compared with groups C and H2,the cell viability was significantly decreased,and the apoptotic rate and concentration of HMGB1 in the supernatant were significantly increased in groups LPS and LPS + H2 (P ＜ 0.05).Compared with group LPS,the cell viability was significantly increased,and the apoptotic rate and concentration of HMGB1 in the supernatant were significantly decreased in group LPS + H2 (P ＜ 0.05).Conclusion Hhydrogen gas can effectively reduce LPS-induced apoptosis in HUVECs through inhibiting the release of HMGB1.

Objective:To observe the effects of community traditional Chinese medicine (TCM) health management of patients with chronic obstructive pulmonary disease (COPD).Methods:From June 2017 to June 2018, 79 patients were selected as study subjects. These patients were randomly divided into two groups. The TCM management group had 39 patients (of which 26 were male), with an average age of (70.77±5.91) years and the course of disease of (7.97±3.12) years. The control group had 40 patients (of which 25 were males), with an average age of (70.60±5.93) years and the course of disease of (7.70±3.01) years. Both groups were given conventional western medicine treatment. The TCM management group was administered TCM health management for 1 year on the basis of the conventional western medicine treatment. This management included establishing electronic archives for COPD patients, Hu Xi Tu-Na, Baduanjin, acupoint massage, acupoint plaster, and dietary guidance according to the TCM physique identification. Management and follow-up of patients was continuous. The TCM syndrome score, CAT score, number of acute exacerbations, and lung function of COPD patients were recorded before and after 1 year of treatment.Results:In the TCM management group, there were 6 cases of clinical control, with 8 and 19 cases displaying obvious and effective effects, respectively. The total effective rate was 84.61%. In the control group, there were 2 cases of clinical control, with 3 cases and 25 cases displaying obvious and effective effects, respectively. The total effective rate was 75.00%. The curative effect of the TCM syndrome in the TCM management group was better than that in the control group. In the TCM management group，the CAT score was (7.46±3.28) points and the number of acute exacerbations was (4.21±2.61) times, the forced expiratory volume (FEV 1) in the first second was (1.99±0.79) L, and the forced vital capacity (FVC) was (2.63±0.92) L. In the control group, the CAT score was (10.38±4.68), the number of acute exacerbations was (6.03±3.00), FEV 1 was (1.66±0.65) L, and FVC was (2.41±0.81) L. The CAT score and the number of acute exacerbations in the TCM management group decreased, while the number of FEV 1 increased, which was statistically significant compared to the control group. Conclusion:Community TCM health management can improve the symptoms of COPD patients at a stable stage, reduce the number of acute exacerbations, and improve lung function.

OBJECTIVE To investigate the improvement effects of glycyrrhizin （GL） on Helicobacter pylori （HP）-associated gastritis in rats and its mechanism. METHODS HP-associated gastritis rat model was induced by inoculating with 1×109 cfu/mL HP. The model rats were randomly divided into model group， positive control group （HP standard quadruple group）， GL low-dose， medium-dose and high-dose groups （5， 20， 50 mg/kg）， with 12 rats in each group. Another 12 healthy rats were selected as normal control group. Except the normal control group and model group were given constant volume of normal saline intragastrically， the other groups were given corresponding drugs intragastrically， once a day， for 30 consecutive days. After administration， rats received 13C urea breath test， and delta-over-baseline （DOB） was recorded； the pathological and cellular morphological changes of gastric mucosa in rats were observed， and pathological scoring was performed； the levels of interleukin-8 （IL-8）， IL-1β， tumor necrosis factor-α （TNF-α）， reactive oxygen species （ROS） and malondialdehyde （MDA） were detected in gastric mucosa of rats； mRNA expressions of high mobility group box-1 protein （HMGB1） and nuclear factor-κ-B （NF-κB）， relative expressions of nitric oxide synthases （iNOS） and HMGB1， the phosphorylation level of NF- κBp65 were also detected in rats. RESULTS Compared with normal control group， the DOB value， histopathological score of gastric mucosa， the levels of IL-8， IL-1β， TNF-α， ROS and MDA， relative expressions of HMGB1 and NF- κB mRNA， relative expressions of iNOS and HMGB1 protein and the phosphorylation level of NF-κB p65 were all increased significantly in model group （P＜0.05）； the epithelial cells of gastric mucosa in rats were incomplete in structure and decreased in the number， with an increase in cell fragments and vacuoles， and significant cell pyknosis. Compared with model group， the changes of the above indexes in GL groups and positive control group were significantly reversed （P＜0.05）； the changes in the above indicators in the GL high-dose group were more significant than GL low-dose and medium-dose groups （P＜0.05）； the pathological changes of gastric mucosal cells in rats had all improved. CONCLUSIONS GL may inhibit inflammation and oxidative stress by inhibiting the activation of HMGB1/NF-κB pathway， thus relieving HP-induced gastric mucosal injury.