Objective To research the mechanism of High Volume Hemofiltration (HVHF) on acute lung injury (ALI) induced by LPS in dogs. Methods After injection of LPS (650 ?g/kg) via central vein within 30 min, Sixteen healthy hybrid male dogs were divided into control group and treatment group randomly (n=8). PaO2、PaCO2 in artery blood were recorded. Contents of TNF-?、IL-6 and IL-10 in plasm were measured by radioimmunity. The activity of NF-?B in lung homogenate was measured by flow cytometer. The content of surfactant protein B (SP-B) in lung homogenate was measured by Western-blotting.Changes of lung histopathology was observed via electron microscopy. Results After injection of LPS, PaO2 and PaO2/FiO2 began to decrease. PaO2 and PaO2/FiO2 in treatment group kept higher than that in control group (P

ObjectiveTo investigate the protective effect of transfected microRNA-146a (miR-146a) on mice with sepsis-induced acute lung injury (ALI) in vivo.Methods Twenty-four healthy male BALB/C mice were randomly divided into sham group, sepsis group, transfection group and transfection control group, eachn = 6. Mice in transfection group were given miR-146a agomir loaded by in vivo-jetPEITM via airway before reproduction of model, and mice in transfection control group were given negative control loaded by in vivo-jetPEITM only via airway. The septic model was reproduced by cecal ligation and puncture (CLP) 12 hours after transfection , and the mice in the sham group underwent laparotomy and closure only without ligation and puncture of the cecum. The mice of each group were sacrificed at 24 hours post-operation. The expression of miR-146a in lung tissue was determined by real time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and the quantity of tumor necrosis factor-α (TNF-α) in the bronchial alveolar lavage fluid (BALF) was determined with enzyme-linked immunosorbent assay (ELISA). The wet/dry ratio of lung (W/D) was determined. The pathohistological changes in the lung were observed and scored. Results The expression of miR-146a showed a significant increase in sepsis group, transfection group and transfection control group, which were (3.56±0.43), (27.64±3.46) and (3.72±0.54) folds of that in sham group, respectively (P< 0.05 orP< 0.01). The miR-146a expression in transfection group was significantly increased compared with sepsis group and transfection control group (bothP< 0.01), but no statistical difference in the expression was found between sepsis group and transfection control group (P> 0.05). Compared with the sham group, higher level of TNF-αin the BALF was found in the sepsis group, transfection group and transfection control group (ng/L: 511.65±43.47, 305.74±34.76, 492.27±42.21 vs. 50.72±7.23, allP< 0.01). The level of TNF-α in transfection group was significantly lower than that in sepsis group and transfection control group (bothP< 0.01). Compared with the sham group, the W/D ratio of lung in sepsis group, transfection group and transfection control group showed a significant increase (6.11±0.32, 5.02±0.29, 6.05±0.43 vs. 4.18±0.10, allP< 0.01). The W/D ratio of lung in transfection group was significantly lower than that of sepsis group and transfection control group (bothP< 0.01). The lung injury score of transfection group was significantly lower than that of sepsis group and transfection control group (6.12±0.75 vs. 10.53±1.52, 9.73±1.08, bothP< 0.01).Conclusions miR-146a agomir loaded by in vivo-jetPEITM instillation into airway was able to increase the expression of miR-146a in the lung tissue of septic mice. Up-regulation of miR-146a inhibit the release of the inflammatory cytokine TNF-α stimulated by sepsis, and alleviate inflammatory reaction and lung tissue injury in mice with sepsis-induced ALI.

ObjectiveTo observe the effect of acetylcholine (ACh) on lipopolysaccharide (LPS) induced inflammatory model of rat alveolar macrophages, and to observe the effect of the acetylcholinesterase inhibitor physostigmine (Phy) on the anti-inflammatory effect of ACh.Methods The rat alveolar macrophages NR8383 were cultured in vitro, which were divided into five groups: blank control group, LPS group (stimulated with 1 mg/L LPS for 12 hours), LPS+ ACh group (0.01, 0.1, 1, 10, 100μmol/L of ACh were added for 5 minutes before LPS stimulation), LPS+ Phy group (1 mmol/L Phy was added for 5 minutes before LPS stimulation), and LPS+ ACh+ Phy group (1 mmol/L Phy and 10μmol/L ACh were added for 5 minutes before LPS stimulation). The supernatants were collected in each group, the enzyme-linked immunosorbent assay (ELISA) was used to assay the contents of tumor necrosis factor-α (TNF-α), interleukins (IL-1β, and IL-6). The activity of acetylcholine esterase (AChE ) in the supernatant was also determined.Results① The contents of TNF-α (ng/L: 605.09±57.13 vs. 34.07±8.62), IL-1β (ng/L: 377.09±28.55 vs. 32.33±10.62) and IL-6 (ng/L: 558.04±77.45 vs. 42.62±11.21) in the LPS group were significantly higher than those in the blank control group (allP< 0.05). These results indicated that the inflammatory model of rat alveolar macrophages was constructed successfully.② ACh with the final concentrations of 0.01, 0.1, and 1μmol/L had less influence on the production of TNF-α, IL-1β and IL-6 in the culture supernatants of alveolar macrophages stimulated with LPS compared with LPS group (allP> 0.05). Nevertheless, 10μmol/L and 100μmol/L ACh notably reduced the production of TNF-α (ng/L: 451.19±30.67, 332.19±32.19 vs. 604.96±22.56), IL-1β(ng/L: 261.08±24.78, 143.98±28.39 vs. 367.06±10.44) and IL-6 (ng/L: 342.75±54.60, 235.48±29.75 vs. 562.69±63.34) in the culture supernatants compared with the LPS group (allP< 0.05).③ The activity of AChE in the LPS group was significantly higher than that in the blank control group (kU/L: 5.21±0.63 vs. 3.09±0.10,P< 0.05). The activity of AChE was successfully inhibited by 1 mmol/L acetylcholinesterase inhibitor Phy pretreatment compared with that in the LPS group (1.51±0.12 vs. 5.21±0.63,P< 0.05).④ The level of TNF-α (ng/L: 183.17±35.44 vs. 451.19±30.67), IL-1β (ng/L: 91.49±12.27 vs. 261.08±24.78) and IL-6 (ng/L: 108.17±22.82 vs. 342.75±54.60) in the culture supernatants of LPS+ ACh+ Phy group was significantly decreased as compared with LPS+ ACh group (allP< 0.05).Conclusions ACh with the final concentrations of 10μmol/L and 100μmol/L can inhibit the LPS induced inflammatory reaction in alveolar macrophages. The acetylcholinesterase inhibitor Phy can reinforce the ACh-mediated anti-inflammatory effect on alveolar macrophages inflammatory model.

AIM: To study the activation effects of maize pollen polysaccharides(PPM) on human thoracic cavity macrophage (hTM). METHODS: Activities of lactate dehydrogenase(LDH) and acid phosphatase (ACP) in the hTM were detected by automatic biochemical analyzer, and the level of tumor necrosis factor alpha(TNF-?) and interleukin-6(IL-6) in the hTM was analyzed by radioimmunoassay, after hTM were cultured with 0.312,0.625, 1.250, 2.500, 5.000 mg/mL PPM-RPMI 1640 for 24 and 48 hours in vitro. RESULTS: The activities of LDH and ACP increased in the hTM induced by PPM, and the levels of TNF-? and IL-6 in the hTM induced by PPM increased markedly too. And the induced expression effect of TNF-? and IL-6 is associated with the concentration of PPM, and time for PPM inducing. CONCLUSION: PPM can induce cytokines secretion in hTM,and activate hTM in vitro.

Objective To evaluate the efficacy of early continuous high-volume-hemofiltration in the treatment of patients with severe acute pancreatitis (SAP).Methods Based on the method of prospective,randomized and controlled clinical trial,60 patients with SAP between January 2005 and July 2011 from the First Affiliated Hospital of Nanchang University were divided into control group and hemofiltration group.The hemofiltration group was treated with early continuous high-volume-hemofiltration and not in the control group.The changes of vital signs,clinical symptoms and laboratory indicators were compared between the two groups before and after the treatment.Results After hemofiltration,the clinical symptoms such as abdominal pain,fever,tachycardia and respiratory distress in hemofiltration group were significantly remitted compared to those in the control group (P ＜0.05).The APACHE Ⅱ score (13.3 ± 1.0 vs 14.1 ± 1.2) and the level of TBil[(20.4±11.3) μmol/L vs (28.1 ±10.9) μmol/L],creatinine[(178.7 ±71.8)μmol/L vs (215.6 ± 51.3) μmol/L],blood urea nitrogen[(10.1 ± 5.6) mmol/L vs (13.2 ± 3.8) mmol/L] and ALT[(51.3 ± 13.2) U/L vs (62.5 ±14.3) U/L] were decreased compared to those in the control group (all P values ＜0.05).The level of PaO2/FiO2(197.3 ±32.4 vs 178.3 ±31.7) was increased (P ＜ 0.05).After hemofiltration,heart rate was decreased gradually (P ＜ 0.05) in the hemofiltration group than in the control group.Mean artery pressure (mAP) increased gradually (P ＜ 0.05) in the hemofiltration group than in the control group.Conclusion Early continuous high-volume-hemofiltration has significant effects on the treatment of SAP including the improvement of clinic symptoms,the blockade of development from systemic inflammatory response syndrome (SIRS) to multiple organ dysfuction syndrome(MODS),improvement of organ function and prevention from the complications.It may become one of the important therapies for SAP.

Objective To explore the mechanism and effect of miR-146a on alveolar macrophages and to observe the changes of miR-146a expression in the LPS-induced alveolar macrophages. Method NR8383 alveolar macrophages were divided into LPS-stimulated group and control group, and the cells of former group were treated with LPS ( 1 μg/mL) and then incubated for 3 h, 6 h and 12 h, respectively. The level of TNF-α in the supernatant of cells was assayed by using enzyme-linked immunosorbent assay (ELISA), and the expression of miR-146a of cells was detected by using Real-Time PCR (TaqMan probe).Statistical analysis carried out by using SPSS 13.0 software package in which One-way ANOVA and Student's t-test were used. Results Compared with control group, the levels of TNF-α in the supernatant of cells were significantly increased 3 h, 6 h and 12 h after LPS challenge (P ＜ 0.01 ). The expression of miR-146a increased 6 h and 12 h after LPS stimulation in NR8383 cells( P ＜0.01 ), and it had an upward tendency.Conclusions The expression of miR-146a in alveolar macrophages increases after LPS-stimulation. It hints miR-146a may be involved in the regulation of the inflammatory responses produced by alveolar macrophages.

Objective To study the effects of recruitment maneuver (RM) strategy on respiratory mechanics and extravascular lung water index (EVLWI) in patients with ARDS. Method Thirty patients with ARDS were randomly divided into RM group and non-RM group. In the RM group, the patients were stabilized with basic mechanical ventilation support for 30 minutes, and then the RM was carried out and repeated once every 12 hous for 3 days. In the non-RM group, patients were supported with mechanical ventilation without RM. The variables of oxygenation index (PaO2/FiO2), peak inspiration pressure (PIP), plateau pressure (Pplat), static pulmonary compliance (Cst) and EVLWI of patients in both groups were determined before treatment and 12 h,24 h, 48 h and 72 h after treatment, and were compared them between two groups. The hemodynamic changes were monitored before and after RM.One-way ANOVA, t -test and Fisher probabilities in 2/2 table were used to process the data. Results ( 1 ) The PaO2/FiO2 and Cst in two groups showed upward trend after treatment, but they were higher in RM group than those in non-RM group ( P ＜ 0. 05 ). The PIP and Pplat of two groups both had downward trend after treatment, but they were significantly lower in RM group than those in group non-RM (P ＜0.05). (2) The EVLWI of two groups showed downward trend after treatment ( P ＜ 0.05), and the differences were significant at all intervals (F: 22.392, 8.147, P ＜ 0.01). The EVLWIs in group RM were lower than those in group non-RM at the intervals of 12 h,24 h, 48 h and 72 h separately (P ＜0.05 or P ＜ 0.01). (3) There were transient hemodynamic changes occurred during RM, and compared with pre-RM, the changes were significantly different ( P＜ 0.01 ). Compared with pre-RM, the hemodynanic changes were not significantly different 120 seconds after the end of RM ( P ＞ 0.05). Conclusions RM could reduce the EVLWI, increase oxygenation and lung compliance.The effect of RM on hemodynamics was transient.

Objective To systematically evaluate the comprehensive effect of subglottic secretion drainage (SSD) on patients with mechanical ventilation (MV) in intensive care unit (ICU). Methods The randomized controlled clinical trials (RCTs) comparing SSD (intervention group) versus non-SSD (control group) in adult patients with MV in ICU was collected through the databases such as the PubMed database of the National Library of Medicine, CNKI, Wanfang database and the Chinese journal of science and technology database (VIP). The subjects were ICU patients with MV, and the retrieval time ranged from January 2006 to December 2016. Two reviewers independently screened the studies according to the inclusive and exclusive criteria, extracted the data, and assessed the quality. Then RevMan 5.3 software was used for Meta-analysis. Sensitivity analysis was performed using Stata 11.0 software. Funnel plot was used to analyze publication bias. Results In the 1004 documents obtained from preliminary screening, a total of 13 studies involving 2052 patients were enrolled after excluding duplicated documents and literature did not meet the inclusion criteria, with 1021 patients in intervention group, and 1031 in control group. Meta-analysis showed that compared with control group, the application of SSD in patients with MV could contribute to the reduction of the incidence of ventilator-associated pneumonia [VAP; risk ratio (RR) = 0.54, 95% confidence interval (95% CI) = 0.46-0.64, P < 0.00001], the duration of MV [mean difference (MD) = -3.29, 95%CI = -4.53 to -2.05, P < 0.00001] and length of hospital stay (MD = -4.27, 95% CI = -7.36 to -1.18, P = 0.007) were shortened, while there was no significant difference in ICU or hospital mortality rate between the intervention group and control group (RR = 0.89, 95%CI = 0.73-1.09, P = 0.25). The sensitivity analysis for studies enrolled in Meta-analysis of MV duration showed that individual research results were stable through step remove of the included literatures and combined calculation of the remaining literature value, suggesting that individual research results were stable, and would not have a significant impact on the overall results. The results of the funnel analysis showed that there was a symmetry in the inclusion studies, and no significant publication bias was found. Conclusions SSD did have effect in reducing the incidence of VAP, shortening the duration of MV and length of hospital stay, while there was no significant effect on reducing mortality rate. Effective use of SSD is an important measure to prevent VAP. It is necessary to objectively evaluate the clinical effect of SSD.

Objective To study the treatment effect and the mechanism of high volume hemofiltration(HVHF)on acute lung injury(ALl)induced by endotoxin in dogs.Methods Sixteen healthy hybrid male dogs were injected LPS(650?g/kg)via central vein within 30 minutes.After model establishment,all animals were divided into two groups randomly( n=8).One group received the treatment of HVHF,while another group received routine treatment.PH,PaO_2,PaCO_2 in arterial blood were recorded at O h after LPS model establishment and 4h after HVHF.Contents of TNF-?,IL-6,and IL- 10 in plasma were measured by radioirnmunity,mRNA expression of TNF-?,IL-6,and IL-10 in lung tissue homogenate were measured by RT-PCR and NF-?B activity by flow cytometer.Results After injection of LPS,PaO_2 and PaO_2/FiO_2 began to decrease,and PaO_2/FiO_2 value was

Objective:To investigate the effect and mechanism of exosomes derived from human-induced pluripotent mesenchymal stem cells (iMSC-Exos) on alveolar macrophages (AM) pyroptosis.Methods:The exosomes in the culture supernatant of human-induced pluripotent mesenchymal stem cells (iMSC) were extracted by rotating ultrafiltration, and the extracted exosomes were identified by transmission electron microscopy, Western blotting and high-resolution adjustable resistance pulse. The rat alveolar macrophage cells (NR8383 cells) were cultured in vitro and the logarithmic growth phase cells were divided into three groups: the control group was added with an equal volume of phosphate buffered saline (PBS) in the AM supernatant; in LPS/ATP group AM cells were stimulated with 500 μg/L LPS for 23 hours and then 5 mmol/L ATP was added for 1 hour to induce pyrolysis; iMSC-Exos group was incubated with AM and 100 mg/L iMSC-Exos for 3 hours before giving LPS and ATP. The cytotoxic activity was detected by cell counting kit-8 (CCK-8) and lactate dehydrogenase (LDH) analysis, the apoptosis and the expression of caspase-1 were observed by immunofluorescence, the levels of inflammatory factors interleukins (IL-1β and IL-18) released by AM were detected by enzyme linked immunosorbent assay (ELISA), the NOD-like receptor protein 3 (NLRP3) inflammasome pathway and the expression level of pyroptosis related protein gasdermin D (GSDMD) were detected by Western blotting. Results:The extracted exosomes were observed by transmission electron microscopy as round vesicles, expressing exosomal markers CD63 and CD9 showed by Western blotting, high-resolution adjustable resistance pulse showed the average diameter of the particles was 130 nm, and could be uptaken by AM. Compared with the control group, the cell activity decreased [(0.56±0.05)% vs. (1.06±0.07)%, P < 0.01], the release of necrotic substance LDH increased (U/L: 1 218.86±22.73 vs. 188.30±1.61, P < 0.01), the expression levels of inflammatory factors increased [IL-1β (ng/L): 958.91±32.78 vs. 194.63±5.14, IL-18 (ng/L): 870.89±21.86 vs. 288.85±24.48, both P < 0.01], and the apoptosis rate [(55.35±6.19)% vs. (12.01±1.32)%, P < 0.01] and caspase-1 expression (fluorescence intensity: 41.06±3.65 vs. 2.80±0.54, P < 0.01) elevated in the AM after LPS/ATP stimulation, suggesting that LPS combined with ATP successfully induced alveolar pyroptosis. Compared with the LPS/ATP group, AM pretreated with iMSC-Exos showed increased cell viability [(0.81±0.05)% vs. (0.56±0.05)%, P < 0.01], decreased LDH secretion (U/L: 535.05±42.55 vs. 1 218.86±22.73, P < 0.01), decreased expression of inflammatory factors [IL-1β (ng/L): 381.82±19.50 vs. 958.91±32.78, IL-18 (ng/L): 533.77±31.54 vs. 870.89±21.86, both P < 0.01], and decreased apoptosis rate [(19.74±2.96)% vs. (55.35±6.19)%, P < 0.01] and caspase-1 expression (fluorescence intensity: 12.16±1.31 vs. 41.06±3.65, P < 0.01). At the same time, the expression of NLRP3 inflammasome pathway [NLRP3 protein (NLRP3/β-actin): 0.62±0.06 vs. 1.89±0.11; cleaved caspase-1 protein (cleaved caspase-1/β-actin): 0.42±0.07 vs. 1.22±0.17, both P < 0.01] and pyrolysis-related protein was significantly inhibited [GSDMD protein (GSDMD/β-actin): 0.57±0.05 vs. 1.22±0.05, P < 0.01]. Conclusion:iMSC-Exos successfully reversed the AM pyroptosis and inflammatory factor expression induced by LPS/ATP, which may be due to the targeted inhibition of NLRP3 inflammasome pathway, suggesting that iMSC-Exos can exert anti-inflammatory effects by inhibiting the pyrolysis of AM.