Objective:To investigate the effect of recombinant Elafin on A549 apoptosis induced by paraquat and the underlying mechanism.Methods:pEGFP-C1-Elafin was transformed into A549 competent cells by electroporation.Transformed A549 was cultured for additional 24 h before Elafin mRNA was detected by RT-PCR and cocultured with or without various concentration of paraquat( PQ ) for different duration.A549 apoptosis percentage and reactive oxygen species ( ROS ) content were tested by flow cytometry .Nuclear factor erythroid like-2(Nrf2) heme oxygenase-1(HO-1) were examed by Western blot .Results:Elafin expressed successfully in A549 after transformation.PQ caused A549 apoptosis in a concentration dependent manner and leaded to ROS content increasing and Nrf2 and HO-1 decreasing.However,elafin could inhibit ROS production and A549 apoptosis,upregulate Nrf2 and HO-1.Conclusion:Elafin could restrict A549 apoptosis to some extent and the possible mechanism lied in its ability to upregulate Nrf2 ex-pression.

Objective To select valuable biomarkers for diagnosis and predicting outcome of sepsis-related acute respiratory distress syndrome(ARDS) from D-dimmer (DD),yon Willebrand factor (vWF),platelet (PLT),N terminal-pro brain natriuretic peptide (NT-ProBNP),interleukin-6 (IL-6),interleukin-8 (IL-8) and surfactant protein D (SP-D).Methods A total of 48 sepsis accompanied with ARDS patients and 40 sepsis patients were prospectively studied with comparison.The clinical characteristics of all the patients were recorded in detail.The blood samples were obtained within 24 hours of ICU admission.The concentration or activity of the seven biomarkers was quantitatively assayed and the results were recorded.To select the most valuable biomarkers as clinical indices,diagnosis model and death predictive model were constructed by Logistic regression.Results Among the seven candidate biomarkers,SP-D,vWF and IL-8 showed the most value.Their area under the receiver operator characteristic curve (ROC) were 0.758 (P ＜ 0.01),0.783 (P ＜ 0.01) and 0.747 (P ＜ 0.01) respectively,and raised to 0.847 (P ＜ 0.001) when the three biomarkers were combined.IL-8,age greater than or equal to 60 years and APACHE Ⅱ score greater than or equal to 20 were related to ARDS death with 12.138(lnIL-8)(P=0.022),6.157(P=0.040) and7.415(P=0.014) of OR values respectively.Conclusion SP-D,vWF,IL-8 should be valuable for early prediction of sepsis-induced ARDS and the diagnostic accuracy raised through combined utilization.IL-8 may be predictable for prognosis of sepsis related ARDS and the comprehensive evaluation combining clinical indices with IL-8 should be suggested in clinical practice.

Objective To investigate the prevalence of GJB2 ,SLC26A4 and mitochondrial DNA 12S rRNA m .1555A>G(mtDNA 1555A>G) mutations in Hui ethic group patients with nonsyndromic hearing loss (NSHL) from Northwest China .Methods A total of 420 peripheral blood samples were collected from unrelated Hui ethic group probands with NSHL in Northwest China .Amplified the target gene by polymerase chain reaction (PCR) af-ter extracting genomic DNA from whole blood .The mtDNA 1555A>G mutation was detected by PCR -Alw26I di-gestion ,then direct sequencing was used to the positive samples of mtDNA 1555A> G ,the coding region of GJB2 gene ,exon 8 and 19 of SLC26A4 gene .Results There were 11(2 .62% ) cases caused by mtDNA 1555A>G homo-zygous mutation in 420 patients with NSHL .There were 41(9 .76% ) cases including homozygote and compound het-erozygote ,caused by GJB2 gene mutation ,which was the most frequent deafness -related gene .The allel frequency of c .235delC accounted for 6 .90% ,as well as the most frequent(51 .33% ) mutational pattern in GJB2 gene .There were 20 patients(4 .76% ) were found carring two allel mutations in SLC26A4 gene .The allel frequency of c .919 -2A>G was 5 .0% ,accounting for a total of 68 .85% in all base alterations of SLC26A4 gene ,which was the major mutant form of SLC26A4 gene .Conclusion GJB2 gene is the most common deafness -gene in Hui ethnic group pa-tients with NSHL from Northwest China ,while c .235delC is the main mutant form ,and c .919-2A>G is the hot-spot mutation of SLC26A4 gene .Through this study we can provide the molecular epidemiology basis for Hui ethnic group patients with NSHL from Northwest China in genetic diagnosis ,genetic counseling and therapy by associated testing of three frequent hearing loss genes .

Objective To investigate the prognostic significance of serum vascular endothelial cadherine (VE-Cad) in patients with acute respiratory distress syndrome (ARDS) induced by sepsis . Methods A prospective observational study was performed between June 2015 and Dec 2017 ,and 48 ARDS patients induced by sepsis from intensive care unit (ICU) were enrolled .And 30 healthy volunteers were enrolled as control .ARDS group was divided into mild group (n=17) ,moderate group (n=18) and severe group (n= 13) .The dynamic levels of serum VE-Cad ,tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were monitored at day 1 ,3 and 7 of admission .Clinical data including extravascular lung water index (EVLWI) ,pulmonary vascular permeability index (PVPI) ,lung injury score (LIS) , APACHEⅡand SOFA were also collected .The t-test or chi square test were used in the comparison between the two groups .One-way ANOVA was used for comparison among multiple groups .Results The serum VE-Cad level of septic group was higher than control group at day 1 of admission ([5 .67 ± 0 .29] vs [0 .28 ± 0 .03] μg/L ,t= 101 .2 , P< 0 .01) .The serum VE-Cad levels in the mild group , moderate group and severe group were (1 .52 ± 0 .59) ,(3 .45 ± 0 .68) ,and (4 .68 ± 0 .53) μg/L , respectively (F=15 .45 ,P<0 .01) .There were positive correlation between VE-Cad levels and EVLWI , PVPI ,LIS ,TNF-αand IL-6 (r=0 .640 ,0 .601 ,0 .507 ,0 .584 ,and 0 .456 ,respectively ,all P<0 .01) . The PaO2/FiO2 and serum albumin level in death group (n=17) were lower than survival group (n=31) ([146 .74 ± 16 .45] vs [245 .42 ± 12 .13] mmHg [1 mmHg=0 .133 kPa] ,t=23 .72 ,P<0 .01 ;[23 .18 ± 3.24]vs[29.16±3.45]g/L,t=5.865,P< 0.01,respectively),andEVLWI ,PVPI,LIS,serum lactate ,mechanical ventilation time ,7 d fluid balance ,APACHEⅡ and SOFA in death group were all higher than survival group .The serum VE-Cad levels at day 1 ,3 and 7 in death group were all higher than survival group ([4 .72 ± 0 .96] vs [3 .36 ± 0 .47]μg/L ,t=8 .801;[3 .87 ± 0 .28] vs [1 .95 ± 0 .42]μg/L , t=16 .86 ;[3 .92 ± 0 .53] vs [0 .96 ± 0 .28]μg/L ,t=25 .42 ,respectively ,all P<0 .01) .The area under curve (AUC ) of VE-Cad for ARDS outcome prediction was 0 .878 with sensitivity of 100 .00％ and specificity of 58 .06％ with a cutoff of 3 .035 μg/L .Conclusion Serum VE-Cad level increases in patients with ARDS induced by sepsis ,and positively correlates with disease severity ,which could be a potential predictor for prognosis .

Platelets are anuclear cell fragments derived from megakaryocytes, playing a critical role in hemostasis and thrombosis through adhesion,activation,release and aggregation. Antiplatelet therapy represents as the cornerstone of secondary prophylaxis of cardiovascular diseases. Increasing evidences sup-port the emerging role of platelets in innate immunity and inflammation and their involvement in the patho-physiological processes of sepsis, atherosclerosis, autoimmune diseases and tumors. Anti-inflammatory and anti-tumor effects of aspirin and P2Y12 receptor antagonist have been reported in clinical trials, suggesting that antiplatelet therapy may have a broad prospect in application. In this review,we focused on the molecu-lar mechanism of platelet involved in immunity and inflammation. In addition,recent findings of antiplatelet therapy for sepsis,atherosclerosis and tumors were discussed.

Objective To detect the expression levels of insulin-like growth factor 1 (IGF-1) and leptin (LP) in serum of patients with colorectal cancer, and to analyze its clinical significance. Methods Seventy-three patients with colorectal cancer, 37 patients with benign colorectal cancer and 40 healthy subjects were enrolled. Serum IGF-1 and LP levels of all the subjects were measured. A two-year follow-up study of colorectal patients was performed to observe the occurrence of lymph node metastasis. Logistic regression was used to analyze the factors affecting lymph node metastasis in colorectal cancer patients. The area under the receiver operating characteristic curve (AUC) was used to analyze the diagnostic value of IGF-1 and LP for colorectal cancer and recurrence. Results The serum IGF-1 concentration in colorectal cancer was significantly lower than that in the control group (P<0.05), but there was no significant difference between the patients with benign lesions (P>0.05). The serum LP level of colorectal cancer was significantly higher than that of benign lesions and control group (P<0.05). Both IGF-1 and LP have certain diagnostic value for colorectal cancer, and the combined test and efficacy of the two methods are higher than those of the separate test (P<0.05). The two-year recurrence rate of colorectal cancer patients was 16.43％, and the patient age (≥40 years old), TNM grade (Ⅲ～Ⅳgrade), tumor diameter (≥4 cm), vascular infiltration and deep muscle infiltration in the recurrent patients were significantly higher than that in the untreated patients (all P<0.05). In the cancer metastasis patients, the level of serum IGF-1 and LP was significantly lower and higher than that in non-metastatic patients, respectively (all P<0.05). Vascular infiltration, deep myometrial invasion, low IGF-1 levels, and high LP levels were independent predictors of recurrence in colorectal cancer patients, and the combined predictive AUC of the above four factors was 0.956 (95％CI:0.9881～0.990). Conclusions In patients with colorectal cancer, IGF-1 level is low and LP level is high, which is closely related to cancer recurrence.

Objective To investigate the prognostic significance of serum VE-cadherin in patients with septic shock. Methods A prospective observation study was performed between January 2016 and December 2017, forty-eight septic shock patients from intensive care unit (ICU) were enrolled, and 25 healthy volunteers served as the controls. Meanwhile, patients in the septic shock group were divided into two subgroups of the survival and death groups according to the 28-day mortality. The dynamic value changes of serum VE-cadherin (VE-Cad), vascular endothelial growth factor (VEGF), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were monitored on the 1st, 3rd and 7th day after admission. Results The serum VE-Cad level of the septic group was significantly higher than that of the control group on the 1st day (3.02±0.18 ng/mL vs. 0.26±0.05 ng/mL, t=3.275, P=0.002). There was a positive correlation between VE-Cad level and VEGF, TNF-α and IL-6 (r=0.826, 0.723, and 0.870, respectively; P<0.01). The PaO2/FiO2 and serum albumin (ALB) level of the death group were lower than that of the survival group, and extravascular lung water index (EVLWI), serum lactate, mechanical ventilation time, 7 day fluid balance, APACHE Ⅱ and SOFA levels of the death group were higher than those of the survival group. The serum VE-Cad levels of the death group were higher than those of the survival group on the 1st, 3rd and 7th day. The serum VE-Cad levels were positively correlated with APACHE Ⅱ and SOFA. The area under the ROC curve of VE-Cad predicting septic shock death on day 1 was 0.723 (95％CI 0.568-0.878). The sensitivity and specificity of VE-Cad with cut-off of 3.1 ng/mL in predicting septic shock death were 60％ and 70.83％, respectively. Conclusions Serum VE-Cad is positively correlated with disease severity and could predict a poor outcome in septic shock patients.

Objective:To evaluate the effects of bosutinib on acute lung injury in mice with endotoxemia.Methods:Sixty clean-grade healthy male C57BL/6 mice, aged 8-12 weeks, weighing 20-25 g, were divided into 4 groups ( n=15 each) using a random number table method: control group (group C), bosutinib group (group B), endotoxemia group (group lipopolysaccharide [LPS]) and bosutinib plus endotoxemia group (group B+ LPS). Septic acute lung injury model was developed by intraperitoneal injection of LPS.Bosutinib 5 mg/kg was injected via the tail vein at 0.5 h before establishing the model in group B+ LPS and at the corresponding time point in group B. At 24 h after developing the model, the mice were sacrificed for microscopic examination of the pathological results of lung tissues which were scored for calculation of the lung coefficient (LI) and wet/dry lung weight (W/D) ratio, and for determination of the content of Evans blue in lung tissues (by Evans blue staining), expression of vascular endothelial cadherin (VE-cadherin), vascular cell adhesion molecule 1 (VCAM-1), phosphorylated nuclear transcription factor κB p65 (p-NF-κB p65), phosphorylated nuclear factor κB inhibitory protein α (pIκB-α) (by Western blot) and expression of interleukin-1beta (IL-1β), IL-6 and tumor necrosis factor-alpha (TNF-α) mRNA (using real-time polymerase chain reaction). Results:Compared with group C, the LI, W/D ratio, Evans blue content in lung tissues and lung injury score were significantly increased, and the expression of IL-1β mRNA, TNF-α mRNA, IL-6 mRNA, VCAM-1, p-NF-κB p65 and pIKB-α was up-regulated, and the expression of VE-cadherin was down-regulated in group LPS ( P<0.05), and no significant change was found in the parameters mentioned above in group B ( P>0.05). Compared with group B, the LI, W/D ratio, Evans blue content in lung tissues and lung injury score were significantly decreased, and the expression of IL-1β mRNA, TNF-α mRNA, IL-6 mRNA, VCAM-1, p-NF-κB p65 and pIKB-α was down-regulated, and the expression of VE-cadherin was up-regulated in group LPS ( P<0.05). Conclusions:Bosutinib can ameliorate the acute lung injury in mice with endotoxemia.

Objective:To evaluate the effect of imatinib on the endotoxemia-induced acute lung injury in mice.Methods:Sixty SPF male, 8-12 weeks, C57BL/6 mice were randomly (random) divided into 4 groups ( n=15 each): control group (group C), imatinib group (groupⅠ), endotoxemia group (group LPS) and imatinib + endotoxemia group (group I+LPS). The endotoxemia model of acute lung injury was established. After 24 hours, the mice were sacrificed. The pathological changes of lung tissues were evaluated, the lung injury scores were calculated, and the wet/dry ratios of lung tissues were measured. ELISA was used to detect the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in serum. Detection kits were used to analyze the levels of malondialdehyde (MDA), glutathione (GSH), catalase (CAT), superoxide dismutase (SOD) and the ratio of reduced glutathione to oxidized glutathione (GSH/GSSG) in lung tissues; The expression levels of phosphorylated nuclear factor-kappa B (p-NF-κB), nuclear factor-erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in lung tissues were analyzed by western blotting. Results:Compared with the group C, the wet/dry (W/D) ratio of lungs [(3.47±0.41) vs. (5.58±0.47)], lung injury scores [(1.25±0.89) vs. (10.25±1.75)], and the levels of TNF-α and IL-6 in serum increased in the group LPS ( P<0.05). The levels of SOD, CAT, GSH and GSH/GSSG decreased, the level of MDA increased, and the expression of p-NF-κB, Nrf2 and HO-1 protein up-regulated ( P<0.05). Compared with the LPS group, the W/D ratio of lungs [(5.58±0.47) vs.(4.62±0.38)] and lung injury scores [(10.25±1.75) vs. (7.00±1.31)] in the I+LPS group decreased ( P<0.05), and the levels of TNF-α and IL-6 in the serum decreased ( P<0.05). In lung tissues, the levels of SOD, CAT, GSH and GSH/GSSG increased (all P<0.05), the level of MDA decreased (all P<0.05), the expression of p-NF-κB protein decreased, and the expression of Nrf2 and HO-1 protein increased ( P<0.05). Conclusions:Imatinib improves sepsis-induced acute lung injury in mice, and the mechanism of actions behind may be related to the inhibition of oxidative stress.

Objective:To evaluate the effects of heparin on focal adhesion kinase (FAK)/Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil-containing protein kinase (ROCK) signaling pathways during acute lung injury (ALI) in septic mice.Methods:Thirty SPF healthy adult male C57BL/6J mice, aged 6-8 weeks, weighing 20-23 g, were assigned into 3 groups ( n=10 each) using a random number table method: control group (group C), ALI group, and heparin group (group H). Septic ALI model was prepared by intraperitoneal injection of lipopolysaccharide 15 mg/kg, while group C received the equal volume of normal saline. In group H, heparin sodium solution 10 U was injected via the tail vein at 30 min before developing the model. The equal volume of normal saline was injected in C and ALI groups. Venous blood samples were collected from the eyeballs under deep anesthesia at 24 h after lipopolysaccharide injection. The mice were subsequently sacrificed and lung tissues were obtained for determination of the serum concentrations of interleukin-1beta (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) (using enzyme-linked immunosorbent assay), wet/dry lung weight (W/D) ratio, expression of vascular endothelial adhesion factor 1 (VCAM-1) (by immunohistochemical staining) and expression of FAK, phosphorylated FAK (p-FAK), RhoA, GTP-bound RhoA (RhoA-GTP) and ROCK (by Western blot) and for examination of the pathological changes. The lung injury was assessed and scored. Results:Comparison with group C, the serum concentrations of IL-1β, IL-6 and TNF-α, W/D ratio and lung injury scores were significantly increased, and the expression of VCAM-1, p-FAK, RhoA-GTP and ROCK was up-regulated in ALI group ( P<0.05). Compared with ALI group, the serum concentrations of IL-1β, IL-6 and TNF-α, W/D ratio and lung injury scores were significantly decreased, and the expression of VCAM-1, p-FAK, RhoA-GTP and ROCK was down-regulated in H group ( P<0.05). Conclusions:The mechanism through which heparin mitigates ALI is associated with the inhibition of the FAK/RhoA/ROCK signaling pathway in septic mice.