Objective To observe the effects of ulinastatin on the expressions of tumor necrosis factor-α (TNF-α) in the lung tissues of rats with acute lung injmy induced by phosgene, and to explore the mechanism of ulinastafin in treating acute lung injury. Method Sixty-four clean grade healthy male SD rots were randomly divided(random number) into eight groups with eight in each group. Group A1 in which rats were exposed to air. Group A2 in which rots wereexposed to air and treated with saline. Group A3 in which rats were exposed to air and treated with dexamethasone. Group A4 in which mrs were exposed to air and treated with ulinastatin. Group B1 in which rots were exposed to phosgene without treatment. Group B2 in which rats were exposed to phosgene and treated with saline. Group B3 in which rats were exposed to phosgene and treated with dexamethasone. Group B4 in which rats were exposed to phosgene and treated with ulinastatin. The expressions of TNF-α in the lung tissues were measured by using immunohistocheistry. Lung tissues were observed grossly and under 200-fold light microscope to identify the positive expressions in kytoplasm. Results In group B1 and group B2, the wet weight, dry weight and wet/dry weight ratio og right lower lobe of lung were higher thai those in group A1 and group A4( P <0.01 ), and those in group B4 and group B3 were significantly lower than those in group B1(P<0.01 ), but still higher than those in group A1 and A4(P<0.01). The gross observation suggested that the surfaces of lung tissues in group A1and group A2 were slick and rose pink without congestion, dropsy or infarction; the surfaces of lung tissue in groups B1 and B2 appeared with congestion, dropsy and many petechia. The surfaces of lung tissue in groups B3 and B4 were similar to those in groups B1 and B2. Under the light microscope, the structure, of lungs in groups A1 and A2 were clear without congestion, effusion or inflammatory cell infiltration. In groups B1 and B2, the engorgement of lung capillary vessels, congestion and tiny thrombosis were found and there abundant edematous fluid and inflammatory cell infiltration in lung stroma and alveolus with focal pulmonary atelectasis in some lung tissue section. The tissues in groups B3 and B4 showed congestion, dropsy, tiny thrombosis and inflammatory cell infiltration, but these changes were slighter than those in groups B1 and B2. The expressions of TNF-α in groups B1, B2, B3, and B4 were significantly higher thanthose in groups A1 and A2( P < 0.01 ), but the expressions of TNF-α IN group B4 was lower than that in groups B1 and B2(P<0.01). Conclusions Ulinastatin could lessen the lung injury by reducing the expressions of pro-inflammatory cytokines such as TNF-α.

OBJECTIVE To investigate the clinical distribution and resistance of Alcaligenes xylosoxidans,discussing risk factors of nosocomial infection and measures of prevention and treatment.METHODS We retrospectively analyzed 1323 isolated strains of Al.xylosoxidans in our hospital from Sep 2004 to Aug 2006.RESULTS For specimen from which Al.xylosoxidans was examined,sputum accounted for 99.85%,being the highest percentage.Al.xylosoxidans was especially distributed at respiratory ward(46.94%) and intensive care unit(ICU) ward(12.77%).Then was cardiovascular ward,endocrinology ward and hematology ward.The antibiotic drugs which had the high drug-resistance rate(i.e.more than 98%) were aminoglycoside(amikacin and gentamicin),quinolone(ciprofloxacin) and cefepime.But the drug-resistance rate to carbapenems,?-lactam/?-lactamase inhibitor combinations and the third generation cepholosporins(cefoperazone and ceftazidime) was less than 5%.CONCLUSIONS It is important for clinic to strengthen the disinfection for hospital environment and to use antibiotic drugs reasonably in order to control colonization and spread of Al.xylosoxidans in hospital.

Objective To investigate the effect of dexamethasone on expressions of angiopoietin-1,2 (Ang-1,2) in rats with acute lung injury induced by phosgene.Methods A total of 36 SD rats were randomly (random number) divided into 3 groups:normal control group that consisted of the rats with air exposure,phosgene group that consisted of the rats with exposure to 8.33 mg/L phosgene (purity 100％,of the same volume as the inhaled air in the normal control group) for 5 minutes and dexamethasone group that consisted of the rats with caudal vein injection of 2.5 mg/kg dexamethasone an hour before exposure to the same dose of phosgene.Wet and dry ratio of the lung (W/D) was calculated,and leukocyte count and total protein content of bronchoalveolar lavage fluid (BALF) were recorded 2 hours later.The concentrations of Ang-1,2 in the serum and BALF were measured by enzyme-linked immunosorbent assay (ELISA).Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the mRNA levels of Ang-1,2 and Tie-2 in the lung tissue.The protein expression of Ang-1,2 and Tie-2 in the lung tissue were quantified by Western blot.Results Compared with phosgene group,the lung W/D,protein content of BALF and WBC count in dexamethasone group were significantly decreased (P ＜ 0.01).Compared with normal control group,Ang-1 and Tie-2 expressions in phosgene group were significantly decreased (P ＜ 0.01).Compared with phosgene group,the serum,BALF and lung tissue of Ang-1 and Tie-2 expressions in dexamethasone group was significantly increased (P ＜0.01).Compared with normal control group,the serum,BALF and lung tissue of Ang-2 expressions in phosgene group were significantly increased (P ＜ 0.01).Compared with phosgene group,the serum,BALF and lung tissue of Ang-2 expressions in dexamethasone group were significantly decreased (P ＜ 0.05,P ＜ 0.01).Conclusion Dexamethasone has a beneficial effect on acute lung injury induced by phosgene in rats by inhibition of Ang-2 and increase in Ang-1 and Tie-2 expressions.

Objective To investigate the role of Wnt/β-Catenin signaling pathway in the endotoxin induced acute lung injury (ALI) during the treatment by mesenchymal stem cells (MSCs).Methods Six SPF male SD rats were isolated and bone marrow mesenchymal stem cells were cultured.A total of 72 SPF male SD rats with 6-week-old were randomly (random number) divided into 4 groups:control group (n =18) in which phosphate buffered solution (PBS) used instead of lipopolysaccharide (LPS);LPS group (n =18) in which LPS used to induce acute lung injury;LPS + MSCs group (n =18) in which MSCs directly transplanted after injection of LPS;Control + MSCs group (n =18) in which MSCs transplanted after injection of PBS.And then 6 rats of each group were sacrificed at 6 h,24 h,and 48 h separately after injection of LPS.At 24 h after the modeling,lung tissue was taken and the levels of Wrnt signaling pathway components were detected by using immunohistochemistry and Western blot.In addition,quantitative realtime PCR was used to detect the expression of Wnt signaling pathway target genes.Results Compare with the PBS control group,significant decrease in lung dry-to-wet ratio and increase in arterial oxygen partial pressure (PaO2) were found in MSCS transplantation groups.According the immunohistological results,Wnt 5a was significantly increased in the LPS-induced ALI rats and decreased after MSCs transplantation.Moreover,decrease in levels of GSK-3β phosphorylation and β-catenin was found in the lung tissue after MSCs transplantation.In addition,the expressions of Wnt signaling target genes Vegf,Axin2 and Klf4 were decreased significantly after MSCs transplantation.Conclusions In the setting of ALI,the therapeutic effect of MSCs was exerted by decreasing the expressions of Wnt 5a,GSK-3β phosphorylation,β-catenin,and Wnt signaling target genes Vegf,Axin2 and Klf4.Wnt signaling implicated in the therapeutic effect of MSC in the setting of ALI.

Objective:To investigate the predictive value of C-reactive protein (CRP), neutrophils-lymphocytes ratio (NLR) and leukocyte-erythrocyte ratio (LER) for aspiration pneumonia (AP) in patients with acute cerebral infarction (ACI).Methods:Retrospective analysis was performed on 989 consecutive hospitalized ACI patients in 2021 who were free of infection within 48 h after ACI onset. General information, past medical history, CRP and complete blood count within 24 h after admission were collected. NLR and LER were calculated based on neutrophil, lymphocyte, leukocyte and erythrocyte count. ACI patients were divided into two groups: non-AP group ( n = 883) and AP group ( n = 106) according to whether they had AP 48 h after admission. Spearman correlations of CRP, NLR and LER with AP were analyzed. The receiver operator characteristic (ROC) curves were plotted to evaluate the predictive values of CRP, NLR and LER for the occurrence of AP in ACI patients, and the sensitivity and specificity at the optimal cut-off value were also calculated. Logistic regression analysis was used for further verification. Results:Compared with the non-AP group, CRP, NLR and LER were significantly higher in the AP group ( P<0.05). Spearman correlation analysis showed that AP was positively correlated with CRP, NLR and LER ( r = 0.42, 0.36 and 0.35, P<0.01). ROC curve analysis showed that CRP, NLR and LER had certain predictive value for AP in ACI patients ( P<0.05), and the area under the curve (AUC) was 0.8917, 0.8349 and 0.8269, respectively. The optimal cutoff values of CRP, NLR and LER were 12.70 mg/L, 4.40 and 1.89 ×10 -3, respectively, with the sensitivity and specificity of 79.25% and 86.41%, 71.70% and 84.94%, and 75.47% and 79.95%, respectively. Multivariate Logistic regression analysis showed that CRP ( OR=6.65, 95% CI: 3.70-11.98, β=1.90, P<0.001), NLR ( OR=2.84,95% CI: 1.60-5.03, β=1.04, P<0.001) and LER ( OR=3.51, 95% CI: 2.00-6.16, β=1.26, P<0.001) were independent risk factors for AP in ACI patients. Conclusions:CRP, NLR and LER at baseline show certain predictive value for the occurrence of AP in ACI patients, and CRP has the strongest predictive power.

Objective: To investigate changes of NLRP3 signal transduction pathway of acute lung injury induced by phosgene to analyze NLRP3-mediated IL-1β release inflammatory process in rats. Methods: Rats were randomly divided into two groups, 10 rats in the Air group that consists of the rats with air exposure, 10 rats in the Psg group that consists of the rats with phosgene exposure at 8.33 g/m³ for 5 min. The specimens of serum, bronchoalveolar lavage fluid (BALF) and lung were collected after 6h. Morphological changes were observed by HE staining. The expression of NLRP3 in the lung of two groups was detected by immunohistochemistry. NLRP3、ASC and caspase-1 expression in the lung tissue was quantified by Western blot. Reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expression of NLRP3、ASC and caspase-1 mRNA in the lung tissue. The concentrations of IL-1β、IL-18 and IL-33 in the serum and BALF were measured by enzyme-linked immunosorbent assay. RT-PCR were used to detect the expression of IL-1β、IL-18 and IL-33 mRNA in the lung tissue. Results: We successfully replicated the model of phosgene-induced ALI in rats. Morphological of HE staining after phosgene exposure to 6 h observed inflammatory cell infiltration in lung tissue in Phosgene group. Immunohistochemical staining results showed that there were many NLRP3 positive cells in lung tissue in Phosgene group. The levels of NLRP3 and caspase-1 mRNA and protein expression in lung were significantly increased (P<0.05) in Phosgene group, but no significant change was observed in lung ASC mRNA and protein expression (P>0.05) . Compared with Air group, the serum, BALF and lung tissue of IL-1β、IL-18 and IL-33 mRNA and protein expression were significantly increased (P<0.01) in Phosgene group. Conclusion NLRP3-mediated inflammatory response probably involved in the process of the phosgene, so it maybe one of the pathogenesis of acute lung injury.

Objective: To investigate the effects of methylprednisolone on NOD-like receptor hot protein domain-associated protein 3 (NLRP3) inflammasome in phosgene-induced acute lung injury. Methods: Rats were randomly divided into four groups, 10 rats in Air group (inhalation of air of the same volume as the phosgene group) , 10 rats in Phosgene group (inhalation of 8.33 mg/L with 100% purity phosgene for 5 min) , 10 rats in Saline control group (inhalation of the same dose of phosgene and 2 mg/kg saline via tail vein injection one hour later) , 10 rats in MP group (inhalation of the same dose of phosgene and 2 mg/kg MP via tail vein injection one hour later) . The specimens of serum, bronchoalveolar lavage fluid (BALF) and lung tissue were collected after 6h. Morphological changes were observed by HE staining. The expression of NLRP3 in the lung of four groups was detected by immunohistochemistry. NLRP3、ASC and caspase-1 expression in the lung tissue was quantified by Western blot. Reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the expression of NLRP3、ASC and caspase-1 mRNA in the lung tissue. The concentrations of IL-1β、IL-18 and IL-33 in the serum and BALF were measured by enzyme-linked immunosorbent assay. Results: We successfully replicated the model of phosgene-induced ALI in rats. Morphological of HE staining after phosgene exposure to 6 h observed inflammatory cell infiltration in lung tissue in Phosgene group. Immunohistochemical staining results showed that there were many NLRP3 positive cells in lung tissue in Phosgene group. The levels of NLRP3, caspase-1 mRNA and protein expression in lung were significantly increased (P<0.05) in Phosgene group compared with Air group; compared with Phosgene group, The levels of NLRP3 and caspase-1 mRNA and protein expression in MP group were significantly decreased (P<0.05) . Compared with Air group, The levels IL-1β、IL-18 and IL-33 mRNA protein expression in the serum and BALF were significantly increased (P<0.05) in Phosgene group. Compared with Phosgene group, The levels IL-1β、IL-18 and IL-33 mRNA protein expression in the serum and BALF were significantly decreased (P<0.05) in MP group. Conclusion Methylprednisolone can effectively protect the rats from phosgene-induced acute lung injury by inhibiting the expression of the NLRP3 inflammasome and reducing the release of inflammatory factors such as interleukin-1β (IL-1β) mediated by it.

Objective To observe the effect of signal transduction pathway of nuclear factor-kappa B (NF-κB) on Nod-like receptor family, pyrin domain-containing 3 (NLRP3) inflammasome and pyroptosis in rats with acute lung injury induced (ALI) by phosgene. Methods The rats were randomly(random number) divided into 3 groups: air exposure control group, phosgene exposure group and PDTC group with phosgene exposure after 100 mg/kg pyrrolidine dithiocarbamate (PDTC) administration. The specimens of serum, bronchoalveolar lavage fluid (BALF) and lung tissue were collected 6 h after exposure. Morphological changes were observed by HE staining. The expression of NLRP3 in the lung of three groups was detected by immunohistochemistry. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expressions of NF-κB p65, NLRP3, ASC and caspase-1 mRNA in the lung tissue. NF-κB p65,NLRP3, ASC and caspase-1 protein levels in the lung tissue were quantified by Western blot. The concentrations of IL-1β, IL-18 and IL-33 in the serum and BALF were measured by enzyme-linked immunosorbent assay (ELISA). Pyroptosis was observed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL). Results The model of phosgene-induced ALI was successfully established in rats. Morphological changes with inflammatory cell infiltration were observed in the lung tissues of phosgene group, in which NLRP3 positive cells also could be observed by immunohistochemical staining. The mRNA expressions and protein levels of NF-κB p65, NLRP3 and caspase-1 in lung tissues were significantly increased (P＜0.05) in phosgene group, compared with air control group. The mRNA expressions and protein levels of NF-κB p65,NLRP3 and caspase-1 in lung tissues were significantly decreased (P＜0.05) in PDTC group, compared with phosgene group. The IL-1β,IL-18 and IL-33 protein levels in serum and BALF were significantly increased (P＜0.05) in phosgene group, compared with air control group. The IL-1β,IL-18 and IL-33 protein levels in serum and BALF were significantly decreased (P＜0.05) in PDTC group, compared with phosgene group. TUNEL results showed that pyroptosis in the lung tissue obviously increased in phosgene group, while decreased in PDTC group. Conclusions NLRP3 inflammasome and lung cell pyroptosis were induced through NF-kB signal transduction pathway in rats with acute lung injury caused by phosgene inhalation. Blockade of NF-κB can alleviate acute lung injury by down-regulating the expression of NLRP3 inflammasome to inhibit pyroptosis.

Objective:To investigate the expression level and clinical value of miR-28-5p in patients with aspiration pneumonia.Methods:This was a retrospective controlled study. A total of 60 patients with severe pneumonia in the Intensive Care Unit of Jinshan Hospital Affiliated to Fudan University were selected as the study group. According to the pathogenic factors, the patients were divided into the aspiration pneumonia group and other infectious pneumonia group. At the same time, 20 healthy physical examination patients in our hospital were selected as the healthy control group. Venous blood was collected from patients in the study group on days 1, 4, and 7. The expression level of miR-28-5p in serum was detected by reverse transcription-polymerase chain reaction (RT-PCR), and the serum levels of interleukin-4 (IL-4), IL-6, IL-8, IL-10, and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA) assay. The clinical detection results of C-reactive protein (CRP) and procalcitonin (PCT) in the study group were collected. Statistical analysis was conducted with SPSS 26.0. Then the diagnostic value of serum miR-28-5p for aspiration pneumonia was analyzed by receiver operating characteristic (ROC) curve, and the correlation between serum miR-28-5p and IL-4, IL-6, IL-8, IL-10, TNF-α, CRP and PCT was analyzed by Pearson method. According to the clinical effect of 10 days of treatment, the patients were divided into the good prognosis and poor prognosis groups, and the relationship between miR-28-5p and the expression levels of various inflammatory factors and prognosis was analyzed.Results:Compared with the healthy control group, the level of serum miR-28-5p in the study group was significantly increased, and the level of serum miR-28-5p in the aspiration pneumonia group was much lower than that in the other infectious pneumonia group ( P<0.05). Compared with day 0, the expression level of serum miR-28-5p in the aspiration pneumonia group was highly increased, with statistical significance ( P<0.05). ROC curve of serum miR-28-5p expression in aspiration pneumonia showed that AUC was 0.871. When the critical value was 1.211, the sensitivity was 76.67% and the specificity was 95%. Pearson correlation analysis showed that miR-28-5p was positively correlated with IL-6 ( P<0.05), and negatively correlated with IL-4 and IL-10 ( P<0.05). ROC curve analysis showed that the area under the curve of age, miR-28-5p, IL-8, IL-10, TNF-α, CRP, and PCT were 0.695, 0.813, 0.655, 0.668, 0.724, 0.651, and 0.661, respectively. Conclusions:Serum miR-28-5p has important reference significance for the diagnosis of aspiration pneumonia, and has certain value for distinguishing different types of aspiration pneumonia. The expression of miR-28-5p in serum is expected to be a new biomarker to judge the prognosis of patients with severe pneumonia. Age, TNF-α, IL-8, IL-10, CRP and PCT are correlated to the prognosis of severe pneumonia.

OBJECTIVETo investigate the dynamic changes of a group of cytokines in phosgene-induced lung injury and the function of different dose of ulinastatin through animal experiment.

METHODS104 male SD rats were randomly assigned into the control group, ulinastatin control group, phosgene treatment groups and different dose of ulinastatin intervention groups, 8 rats each group. Treatment groups were dynamic constant exposure in phosgene, and immediately injected ulinastatin intraperitoneal, and then the experimental animal, the lung tissue biopsy, lung wet/dry ratio, RT-PCR detection, the plasma for detection of Bio-Plex 18 cytokines.

RESULTSCompared with the control group, plasma concentrations of IL-1α, IL-6, GM-CSF, TNF-α, INF-γ, MIP-3α, VEGF were increased significantly first (2 h), and gradually decreased with the passage of time , the difference was statistically significant (P < 0.05). Plasma concentrations of IL-4, IL-10 were decreased earlier (2h, 6 h) and increased later (24 h) (P < 0.05). The change of plasma concentration of IL-13 was not obvious earlier (2 h) and still rising later (24h), the difference was statistically significant (P < 0.05). After drug intervention, the levels of pro-inflammatory cytokines declined and the levels of anti-inflammatory cytokines raise by different degrees at different times in ulinastatin intervention groups, the difference was statistically significant. The degree of lung injury was improved than the phosgene treatment groups and better in high dose of ulinastatin intervention group. The expression of IL-10, IL-4, IL-13-mRNA of tissue increased in accordance with plasma results.

CONCLUSIONA group of cytokines are dynamicly changed in phosgene-induced lung injury by time. High dose of ulinastatin can improved phosgene-induced lung injury, regulate the synthesis and release of inflammatory cytokines and inhibit inflammatory react in a dose-dependent manner.

Animals ; Cytokines ; metabolism ; Glycoproteins ; pharmacology ; Granulocyte-Macrophage Colony-Stimulating Factor ; Interleukin-10 ; Interleukin-13 ; Interleukin-1alpha ; Interleukin-4 ; Interleukin-6 ; Lung ; Lung Injury ; chemically induced ; drug therapy ; Male ; Phosgene ; toxicity ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha