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Objective To explore the impact of glucocorticoid on coagulation through administrating on rats with smoke inhalation.Methods Totally 150 male S-D rats were randomly (random number) divided into 5 groups:control group (ambient air inhalation),smoke group (smoke inhalation for 30 min),smoke+high dosage methyl prednisolone group(MP 40 mg/kg,intraperitoneal injection,s+HMP group),smoke+medium dosage MP (4 mg/kg) group (s+MMP group),smoke+low dosage MP (0.4 mg/kg) group (s+LMP group) (all n=30).Survival rates were calculated 24 h after smoke inhalation.Lung tissues were collected for histopathology and wet to dry (W/D) ratio.Arterial blood was collected for blood gas test.Coagulation factors in lung and plasma were tested.Results Survival rates of three MP groups were markedly improved compared with the smoke group (all P＜0.05),and was significantly higher in the medium dosage group(85.17％) than those in the low and high dosage groups (65.73％ and 60.07％,all P＜0.05).The W/D ratio and blood gas test were markedly improved in the high and medium groups (all P＜0.05).Tissue factor (TF) and thrombin-antithrombin complex (TAT-c) in bronchoalveolar lavage fluid (BALF) increased dramatically after SI (P＜0.01,P=0.005) with a remarkable drop of factor Ⅱ (F Ⅱ) (P=0.007),all of which were attenuated by MP with dosage dependence.The mRNA expression of TF increased dramatically after SI and recovered significantly with MP administration,while the expression of thrombomodulin (TM) recovered in the opposite direction with MP,all of which were in a dosage dependent manner.TF,fibrinogen (FIB),TAT-c increased significantly in plasma after smoke inhalation (P＜0.01,P=0.027,P=0.005).F Ⅷ ％ increased with MP administration and TF was raised by high dosage MP compared with the smoke group.FIB and TAT-c were decreased in all MP groups,which were significant higher in the high and middle dosage groups.The change of TM and endothelial cell protein C receptor (EPCR) in circulation were similar with FIB or TAT-c with or without MP.Protein C (PC％) and antithrombin (AT Ⅲ ％) dropped dramatically after SI,high and middle dosages of MP could restore the activity significantly,while low dosage would restore AT Ⅲ ％ but not PC％.Conclusions Glucocorticoid can significantly improve local and systemical coagulation disorder caused by smoke inhalation,and high-and medium-dosage hormones are effective.The regulation of hormones on the coagulation system is an important mechanism in the treatment of smoke inhalation induced lung injury.
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Objective To investigate the therapeutic effect of different doses of methylprednisolone (MP) in smoke inhalation-induced acute lung injury (SI-ALI).Methods Adult male Sprague-Dawley (SD) rats were divided into control group (group A,n = 6), smoke inhalation group (group B, smoke inhalation 30 minutes,n = 30) and smoke+MP 40, 4, 0.4 mg/kg intervention group (groups C, D, E; intraperitoneal injection of MP at 1 hour before smoke inhalation, n = 30) according to random number table method. The survival status of rats in each group was observed at 24 hours, and murine smoke inhalation induced trauma score (MSITS) according to the symptoms and signs of rats at 3 hours after smoke inhalation were scored. The blood of abdominal aorta of rats was collected. Then the rats were sacrificed to harvest bronchoalveolar lavage fluid (BALF) and lung tissue. The levels of interleukin (IL-6, IL-17a) in plasma and BALF were detected by enzyme linked immunosorbent assay (ELISA); the total number of white blood cells and the proportion of leukocytes or macrophages in BALF were calculated; the histopathological changes of lung were observed and the lung injury score was given; the expression of myeloperoxidase (MPO) and high mobility group protein B1 (HMGB1) in lung tissue were detected by Western Blot.Results The 24-hour survival rate of group B rats was 33.67％. The survivalrate of groups C, D and E (65.73％, 85.17％, 60.07％) were significantly higher than that of group B (allP < 0.05), and the survival rate of group D was significantly higher than that of groups C and E. Diffuse inflammatory cell infiltration, intra-alveolar hemorrhage and a large amount of edema fluid were seen in the lung tissue of group B; and the lung injury score was significantly higher than that of group A. Compared with group B, the lung injury in different doses of MP group were decreased to different degrees, while the lung injury scores in groups C and D were significantly decreased (3.31±1.37, 2.62±0.98 vs. 5.52±0.97, bothP < 0.01); correlation analysis showed that MSITS score was significantly and positively correlated with lung injury score (r = 0.862,P < 0.001). The levels of plasma inflammatory factors and BALF protein, inflammatory cells and inflammatory factors, and the expression of MPO, HMGB1 in group B were significantly higher than those in group A. Compared with group B, the levels of inflammatory factors in plasma, and protein content, inflammatory cells and inflammatory factors in BALF in different doses of MP group were decreased to different degrees, with significant differences in groups C and D [plasma: IL-17a (pg/L): 49.28±27.12, 36.57±16.52 vs. 191.79±88.21; IL-6 (ng/L): 206.47±109.96, 197.52±113.86 vs. 669.00±299.60; BALF: protein content (mg/L):892.0±164.5, 566.1±120.9 vs. 1838.0±145.8; white blood cell count (×109/L): 5.40±1.67, 2.81±1.20 vs. 9.02± 2.06; neutrophil ratio: 0.315±0.081, 0.273±0.080 vs. 0.590±0.096; IL-17a (ng/L): 22.63±8.62, 18.92±8.43 vs. 43.31±19.17; IL-6 (ng/L): 156.49±46.94, 123.66±64.91 vs. 253.43±80.03; allP< 0.01]; in addition, the expression of MPO and HMGB1 protein in lung tissues of MP groups with different doses were significantly decreased, the expression of MPO in group D was significantly lower than that in group E [MPO/β-actin (fold increase from group A):2.14±0.97 vs. 4.35±0.87,P < 0.01], the expression of HMGB1 in groups C and D were significantly lower than that in group E [HMGB1/β-actin (fold increase from group A): 1.77±0.73, 1.23±0.67 vs. 3.65±1.08, bothP < 0.05]. Conclusions MP can significantly improve the survival rate of SI-ALI rats and reduce the acute pulmonary and systemic inflammatory response. The MP effect of 4 mg/kg was better than 40 mg/kg and 0.4 mg/kg.
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Objective To explore the effects of hydrogen sulfide (H2 S) on the inflammatory reaction and the expression of single immunoglobin IL-1 receptor related protein (SIGIRR) in rats with acute lung injury (ALI) induced by lipopolysaccharide (LPS). Methods Forty male SD rats were randomly divided into 4 groups (normal group, H2 S group, ALI group and ALI + H2 S group). The ALI rat model was established by LPS peritoneal injection. After LPS stimulation for 1 h, rats inhaled H2 S 80mg /m3 for 6h. Then, rats were sacrificed with a supraphysiological dose of pentobarbital sodium. The histological changes in the lung, the levels of TNF-α and IL-1β in serum and lung tissue homogenates, and the protein expression of SIGIRR in lung tissues were examined. Results Compared with the normal and H2 S groups, typical histological features of ALI were observed in ALI group, and the lung injury scores of ALI group were higher than those of the normal and H2 S groups (P ＜ 0. 05). Moreover, there were marked increases in the levels of TNF-α and IL-1β in serum and lung tissue homogenates after LPS injection. In contrast, inhalation of H2 S could attenuate lung pathological changes and inhibit the productions of TNF-α and IL-1β in serum and lung tissue homogenates (P ＜ 0. 05). Additionally, inhalation of H2 S could induce the protein expression of SIGIRR in rat lung tissues. Conclusion Inhalation of H2 S protected rats from LPS-induced ALI and its mechanisms were partially associated with inhibition of the productions of TNF-α and IL-1β and modulation of SIGIRR expression.
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Objective To explore the effect of curcumine on the inflammation and expression of single immunoglobin IL-1 receptor related protein in alveolar epithelial cells induced by lipopolysacharride (LPS).Methods The rat type Ⅱ alveolar epithelial cells were cultured in vitro,and cell activity was measured when stimulated with LPS and different doses of curcumin.The level of tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6) in supematant was detected.Cells pretreated with curcumin (20 μmol/L),were stimulated with LPS (10 μg/mL).The nuclear protein and membrane protein was extracted to detect the expression level of nuclear transcription factor kappa B (NF-κB) and single immunoglobin IL-1 receptor related protein (SIGIRR).Results The cells activities were not affected by curcumin (5 ～30 pμmol/L) and LPS (10 μg/mL) (P ＜ 0.05).Curcumin (5 ～30 μ mol/L) significantly inhibited LPS-induced overpression of TNF-α and IL-6 (P ＜ 0.05).In 20 μ mol/L and 30 μ mol/L pretreatment groups,the inhibition of curcumin was most obvious,and there were no significant differences between the two groups (P ＜ 0.05).Curcumin (20 μ mol/L) significantly inhibited the expression level of phosphorylation of NF-κB p65 in cell nucleus,while up-regulated the expression of SIGIRR (P ＜ 0.05).Conclusion Curcumin inhibits the expression of inflammatory factor such as TNF-α and IL-6 as well as activation of NF-κB in alveolar epithelial cells induced by LPS.Up-regulating the expression level of negative regulatory molecules SIGIRR is one of the possible mechanism.
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AIM: To investigate the role of G-protein-coupled bile acid receptor 1 ( GPBR1; also known as TGR5) activation in high glucose-induced cardiomyocyte hypertrophy and calcineurin (CaN)/nuclear factor of activated T-cells 3 (NFAT3) signaling.METHODS:Primarily cultured mouse cardiomyocytes were used in the study .The cell surface areas of the cardiomyocytes were measured by an image analysis system .The cell protein content was detected by BCA meth-od.The expression of TGR5, CaN and NFAT3 at mRNA and protein levels was determined by RT-PCR and Western blot . RESULTS:The mouse cardiomyocytes were successfully cultured .High glucose significantly induced the increases in the cell surface area, the cell protein content and the expression of CaN and NFAT 3 (P<0.05) in the cardiomyocytes.TGR5 activation or a CaN antagonist cyclosporin A inhibited high glucose-induced cardiomyocyte hypertrophy and the expression of CaN and NFAT3 (P<0.05).These effects of TGR5 activation were abolished by TGR5 gene interference (P<0.05). CONCLUSION:TGR5 activation reduces high glucose-induced cardiomyocyte hypertrophy by inhibiting CaN /NFAT3 sig-naling.
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Objectives To explore the effect of the standardized residents training method on intern teaching in department of respiratory. Methods Sixty interns of Grade 2010 from the Second Military Medical School were randomly divided into 2 groups with 30 interns each. The traditional teaching method was adopted in control group, while the standardized residents training method was used in experiment group. When the rotating internship was finished in department of respiratory, the survey of satisfaction about teaching and the assessment of teaching effect were performed in two groups. The contents of examination included academic knowledge exam, clinical skills test, basic skills test and comprehensive quality assessment. Results There was no difference between two groups in academic knowledge exam (P>0.05). The total scores, clinical skills scores and basic skills scores of experiment group were higher than those of control group (P<0.05). And, higher satisfaction was ac-quired in experiment group(P<0.05). Conclusions Using the standardized residents training method can improve the effect of the intern teaching and teaching satisfaction and it can be widely applied in intern teaching.
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Objective To construct the bait plasmid of pSos-single immunoglobin IL-1 receptor related protein (SIGIRR) in Cy-toTrap yeast two hybrid system ,and to test its self-activation .Methods The cDNA fragments of SIGIRR(480 -1 230 bp) were amplified from pReceiver-LV19-SIGIRR and ligated into the bait plasmid pSos to generate the plasmid pSos-SIGIRR .The pSos-SI-GIRR was identified by DNA sequencing and dual-site endonuclease digestion .Then the recombinant plasmid and control plasmid were introduced into the yeast cell cdc25H .The transformants were inoculated on plates of 25 ℃ /SD/Glucose(-UL) ,25 ℃/SD/Ga-lactose(-UL) ,37 ℃ /SD/Glucose(-UL) and 37 ℃ /SD/Galactose ,respectively and the proliferation ability of transformant was ob-served for 6 d .The Western blot was adopted to detect the expression of target protein .Results The pSos-SIGIRR vector was cor-rectly constructed and proved of no self-activation and toxic action .The Western blot showed that the target protein was expressed in a form of fusion protein of 170KD .Conclusion The bait plasmid containing SIGIRR cytoplasmic tail can be applied to the yeast two-hybrid system and lays the important foundation for seeking the interacting protein with SIGRR from the human lung cDNA li-brary in .