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Objective@#To investigate the serum levels of interleukin (IL)-6 and -8 in patients with chronic periodontal disease and acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and the possible relationship between IL-6 and IL-8 with two diseases.@*Methods@#A total of 40 cases of healthy subjects (control group 1) from graduate school of Anhui Medical University, and 120 cases (40 cases in each of the 3 groups) of eligible patients were collected, of which 40 were patients with chronic periodontal disease and AECOPD (experimental group) from Department of Respiratory Medicine, The First Affiliated Hospital of Anhui Medical University and Anhui NO.2 Provincial People's Hospital, 40 were patients with chronic periodontal disease (control group 2) from Department of Stomatology, The First Affiliated Hospital of Anhui Medical University, 40 were patients with AECOPD (control group 3) from Department of Respiratory Medicine, The First Affiliated Hospital of Anhui Medical University. The clinical indicators of all subjects were collected, including tooth mobility degree, probing depth (PD), bleeding index (BI), attachment level (AL), vital capacity max (VC Max), forced expiratory volume in first second (FEV1) and forced expiratory volume in first second to forced vital capacity (FEV1/FVC) ratio. Enzyme linked immunosorbent assay (ELISA) was used to detect the serum levels of IL-6 and IL-8 in the subjects of four groups.@*Results@#The attachment levels had no significant differences between experimental group and control group 2 (P>0.05). The pulmonary function indices of experimental group including VC MAX% pre[(56.1±11.1)%], FEV1 %pre [(44.8±12.2)%], FEV1/FVC(%) [(56.8±11.4)%] were significantly different from those in control group 3 [(66.3±10.1)%, (53.0±10.4)%, (66.5±8.2)%, respectively]. The IL-6 levels of experimental group, control groups 1, 2 and 3 were (14.4±3.9), (2.1±1.1), (4.8±1.9) and (8.6±1.4) ng/L, respectively. And the IL-8 levels were (35.3±33.3), (4.8±1.7), (9.7±3.3) and (15.6±9.6) ng/L. In experimental group the IL-6 and IL-8 levels were significantly higher than those in control groups 1, 2, and 3 (P<0.01). In control group 2 and 3 the IL-6 and IL-8 levels were significantly higher than that in control group 1 (P< 0.01).@*Conclusions@#The IL-6 and IL-8 levels of experimental group were significantly increased. IL-6 and IL-8 may be associated with the development of periodontal disease and AECOPD closely.
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Objective To assess the value of serum procalcitonin (PCT) level in the differentiation between infections and tumor-related fever in patients with malignant tumors.Methods Serum levels of PCT and related indicators were measured in 139 patients (173 times) with malignant tumors admitted in the First Affiliated Hospital of Anhui Medical University from December 2014 to December 2015.There were 64 cases of infection with fever,44 cases of infection without fever,36 cases of tumor-related fever and 29 non-infection,non-fever cases.Serum PCT levels of patients in 4 groups were compared by non-parametric test,and the value of PCT in the differentiation of infections and tumor-related fever was assessed by receiver operating characteristic (ROC) curve.Results There were significant differences in serum PCT levels among patients with different type of tumors (x2 =11.238,P =0.047) and between patients with metastases and without metastases (x2 =12.658,P =0.005) in infected patients,as well as in non-infected patients (x2 =12.374,9.942,P =0.015,0.019;respectively).The PCT levels of infection with fever group,infection without fever group,tumor-related fever group and non-infection,non-fever group were 0.449 (0.148,2.090) μg/L,0.107 (0.056,0.441) μg/L,0.254 (0.100,0.530) μg/L and 0.058 (0.043,0.124)μg/L,respectively.There was no significant difference in PCT levels between tumor-related fever group and infection without fever group (Z =-1.480,P =0.139),between tumor-related fever group and blood culture negative infection with fever group (Z =-0.965,P =0.334).According to the ROC,the cutoff value for PCT was defined at 0.955 μg/L,the sensitivity and specificity of PCT in differentiation between infection and tumor-related fever was 0.34 and 0.94,respectively.Conclusion Tumor types and metastasis can affect serum PCT level,and PCT level can be increased in tumor-related fever,which would affect value of PCT in diagnosis of infection in tumor patients.
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Objective To investigate the possibility of the involvement of RhoA/mDia1 pathway to cause the expression of phosphorylate ezrin-radixin-moesin (p-ERM) in rat pulmonary micro-vascular endothelial cell (PMVEC) after the stimulation of lipopolysaccharide (LPS).Methods The specimens of lung tissue were taken from healthy male SPF grade SD rat with 100-120 g body weight which was purchased from the laboratory animal center of Anhui province.After culture,the PMVECs were randomly divided into dose-dependent groups (0,0.1,1,10 μg/mL LPS added in PMVECs and cultured for30 min,n =8 in each),time-dependent groups (10 μg/mL LPS added to PMVECs cultured for 0,15,30,60,120 min,n =8 in each) and intervention group (n =8).In the intervention group,PMVECs were cultured with 1 μg/mL C3 transferase in serum free media for 240 min,followed by treatment with 10 μg/mL LPS for 30 min.Meanwhile,two control groups in serum-free DMEM medium were made by adding 10.μg/mL LPS to PMVECs and 1 μg/mL C3 transferase to PMVECs respectively cultured for 30 min (n =8 in each).Western blot was used to detect the level of p-ERM,ERM and mDia1.Data were analyzed with SPSS 16.0 software,while one way analysis of variance (ANOVA) was used to compare multiple sets of variables,the intergroup comparisons were analyzed by the least-significant-difference (LSD) tests,with P ＜0.05 for the statistically significant difference.Results ERM,p-ERM and mDia1 were presented in rat PMVEC.Stimulation with LPS up-regulated p-ERM,mDia1 in a dose-dependent manner:LPS [0 μg/mL LPS group:(0.520±0.101),0.1 μg/mL LPS group:(0.657 ±0.092),1 μg/mL LPS group:(0.891 ±0.167),10 μg/mL LPS group:(1.227 ±0.106);0 μg/mL group vs.0.1 μg/mL group,P ＞0.05;the rest P ＜0.01];and mDia1 [0 μg/mL LPS group:(0.200 ±0.102),0.1 μg/mL LPS group:(0.430 ±0.121),1 μg/mL LPS group:(0.603 ±0.154),10 μg/mL LPS group:(0.887 ±0.204);0.1 μg/mL group vs.1 μg/mL group,P ＞ 0.05;the rest P ＜ 0.05].In time-dependent group,the level of p-ERM protein increased at 15 min (0.670 ±0.149),peaked at 30 min (1.175 ±0.103),then decreased,at 60 min (0.959 ±0.189),90 min (0.842 ±0.129),but kept at higher level at 120 min (0.767 ±0.097) than that in control group (0.471 ±0.157,15 min group vs.120 min group,60 min group vs.90 min group and 90 min group vs.120 min group,P ＞ 0.05;the rest P ＜ 0.05);and the level of mDia1 increased at 15 min (0.779 ±0.035),peaked at 30 min (0.889 ±0.036) then decreased at 60 min (0.648 ±0.019),90 min (0.582 ±0.068),but kept at higher level at 120 min (0.526 ±0.059) than that in control group (0.284±0.118,all P ＜ 0.01).C3 transferase caused a marked attenuation of LPS induced p-ERM expression [control group:(0.339 ± 0.069);C3 + LPS group:(0.438 ± 0.07);C3 control group:(0.352 ± 0.071);LPS control group:(0.634 ± 0.191),C3 + LPS group vs.LPS control group,P =0.01],as the same in mDia1 [control group:(0.449 ±0.122);C3 + LPS group:(0.380 ±0.148);C3 control group:(0.404 ±0.164);LPS control group:(0.622 ±0.174),C3 + LPS group vs.LPS control group,P ＜ 0.01].Conclusions LPS could up-regulated the expression of p-ERM protein,and inhibition of RhoA/mDia1 signal pathway by C3 transferase could down-regulated the p-ERM levels.
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Objective To investigate the role of phosphorylated Moesin (p-Moesin) in the injury of pulmonary microvascular endothelial cells (PMVECs) of rats induced by tumor necrosis factor-α (TNF-α), and to approach the impact of Rac1 signal pathway on Moesin phosphorylation.Methods PMVECs of rats were culturedin vitroand passed on to the third generation, and the TNF-α time-effect experiment, dose-effect experiment and Rac1 signaling pathway intervention experiment were performed respectively. ① Time-effect experiment: PMVECs were stimulated with 10μg/L TNF-α for 0, 15, 30 minutes and 1, 3, 6, 12 hours, and the protein expressions of Moesin and p-Moesin were determined by Western Blot. ② Dose-effect experiment: PMVECs were stimulated with 0, 0.1, 1, 10μg/L TNF-α for 6 hours, and the protein expressions of Moesin and p-Moesin were determined by Western Blot. ③ Rac1 signaling pathway intervention experiment: PMVECs were divided into two parts, which were pretreated with 3 mL Rac1 specific inhibitor NSC23766 (200μmol/L) for 0.5 hour or Rac1 specific agonist O-Me-cAMP (200μmol/L) for 1 hour, respectively, and then incubated with 10μg/L TNF-α for 6 hours. The PMVECs without treatment were served as blank control group, andthose were treated with only O-Me-cAMP, NSC23766 or TNF-α were served as corresponding groups. The protein expressions of Moesin and p-Moesin were determined by Western Blot.Results ① Time-effect experiment results: the expression of Moesin showed no change among all time points after 10μg/L TNF-α stimulated PMVECs. But the expression of p-Moesin was sharply up-regulated at 15 minutes after TNF-α stimulation as compared with 0 minute (p-Moesin/Moesin: 4.399±0.523 vs. 1.000±0.195), peaked at 30 minutes (6.069±0.557), and then gradually decreased after 1 hour (5.005±0.544, 4.599±0.478, 1.742±0.288, 1.503±0.352 at 1, 3, 6, 12 hours, respectively) with significant difference among all time points (F = 15.397,P = 0.002). ② Dose-effect experiment results: no significant change in expression of Moesin was found among all doses of TNF-α incubated with PMVECs for 6 hours. But the expression of p-Moesin was significantly up-regulated after TNF-α stimulation with 0.1μg/L as compared with0μg/L (p-Moesin/Moesin: 2.194±0.430 vs. 1.000±0.273), which showed an upward trend with dose increase in TNF-α (3.201±0.688 and 4.413±0.296 with 1μg/L and 10μg/L TNF-α, respectively) with significant difference among all doses (F = 92.513,P < 0.001). ③ Rac1 signaling pathway intervention experiment results: there was no significant difference in Moesin expression among all the groups. Compared with blank control group, Rac1 specific agonist O-Me-cAMP or Rac1 specific inhibitor NSC23766 alone could not change the expression of p-Moesin, while TNF-α could induce p-Moesin expression. Compared with TNF-α group, the expression of p-Moesin induced by TNF-α was up-regulated by NSC23766 (p-Moesin/Moesin: 2.612±0.355 vs. 1.911±0.297,P < 0.05), and it was attenuated by O-Me-cAMP (p-Moesin/Moesin: 1.928±0.331 vs. 3.030±0.353,P < 0.05).Conclusion The phosphorylation of Moesin is involved in the damage of TNF-α-induced PMVECs in rats, and PMVECs damage could be alleviated by modulating Moesin phosphorylation in the Rac1 signaling pathway.
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Objective To assess the accuracy and safety of medical thoracoscopy(MT)in the diagnosis of tuberculous pleural effusion.Methods We evaluated 52 patients who were suspected tuberculous pleural effusion.The diagnosis rate and complications of medical thoracoscopy was assessed.Results About 33 of 52 patients were tuberculous pleural effusion.Twenty-nine cases were diagnosed by medical thoracoscopy,and the diagnostic rate was 88%.Under the thoracoscope,clinical manifestations of these pa-tients with tuberculous pleuritic were miliary nodules in 23 cases (70%),fiber cord-like adhesions in 12 cases (36%),extensive wrapped with fiber deposition in 7 cases(21%),and white scar in 5 cases (1 5%).All complication was relived or caused,and 1 case of gas embolism was the most serious one.Conclusion Medical thoracoscopy was a method with high diagnostic value and safety in the diagnosis of tuberculous pleural effusion.
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Objective To investigate the effect of tumor necrosis factor-α (TNF-α) on the levels of ezrin-radixin-moesin (ERM) proteins and the phosphorylated ERM proteins (p-ERM) in rat pulmonary microvascular endothelial cells (PMVEC),and to explore Rho kinase (ROCK) influencing on modulation of the ERM proteins phosphorylation.Methods Cultured rat pulmonary microvascular endothelial cells were randomly divided into dose-dependent and time-dependent groups.In dose-dependent group,cells were cultured with different doses of TNF-α (0,0.1,1,10 μg/LTNF-α) for 60 min.In time-dependent group,cells were cultured with TNF-α (10 μg/L) for 0,15,30,60,90,120,180 min.In ROCK inhibitor (Y27632) intervention group,cells were cultured with TNF-α (10 μg/L) or Y27632 (30 μmol/L) + TNF-α (10 μg/L) for 60 min respectively.The levels of ERM proteins and p-ERM were determined by western blot.One way analysis of variance (ANOVA) was employed for statistical analysis by using SPSS version 16.0 software to compare values among all groups.A significant difference was presumed as a P value ＜ 0.05.Results Western blot revealed that ERM and p -ERM proteins were present in rat PMVEC.Stimulation withTNF-α gradually up-regulated the level of pERM proteins in a dose-dependent manner [0 μg/LTNF-α group:(0.648 ± 0.102),0.1 μg/LTNF-αgroup:(0.728-±0.082),1 μg/LTNF-α group:(0.926±0.121),10 μg/LTNF-α group:(1.245 ±0.134),all P =0.000].In time-dependent group,the level of p-ERM proteins rose at 15 min (0.777 ±0.151),peaked at 90 min (1.295 ±0.176),then decreased gradually at 120 min (0.802 ±0.139),but stayed higher level at 180 min (0.669 ±0.128) than that in un-stimulated 0 min group (0.631 ±0.123,P=0.004,0.000,0.001,0.016,respectively).When PMVEC pre-incubated with ROCK inhibitor and TNF-t,the level of p-ERM proteins caused a marked attenuation of TNF-αstimulation [(0.634 ± 0.112) vs.(0.875 ± 0.164),P =0.002],however,there are no significant differences compared to ROCK inhibitor alone group (0.661 ± 0.108) and no intervention group (0.654 ± 0.125),P =0.973,P =0.900,respectively).Conclusions TNF-α could induce up-regulation of the level of the phosphorylated ERM proteins in rat PMVEC,and ROCK signal molecules might involve in modulation of the ERM proteins phosphorylation.
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Objective To investigate the diagnostic value of YKL‐40 and CEA in malignant pleural effusion .Methods There were 45 cases of benign pleural effusion and 52 patients with malignant pleural effusion in this study from November 2013 to No‐vember 2014 .Enzyme linked immunosorbent assay(ELISA) and electrochemi lumine scence immunoassay(ECLIA) method were carried out to detect the concentration of YKL‐40 and CEA respectively .The differences of the two groups of patients between YKL‐40 and CEA levels were compared ,and the correlation between YKL‐40 and clinical pathology of malignant pleural effusion were analyzed .In addition ,the receiver‐operating characteristic curve(ROC curve) was used to compare the diagnostic value be‐tween YKL‐40 and CEA .Results The average value of YKL‐40 in malignant pleural effusion was (189 .5 ± 147 .0)ng/mL ,and sig‐nificantly higher than in benign pleural effusion group(P0 .05) .The diagnostic sensitivity and specificity of YKL‐40 was 80 .9% and 51 .2% ,which was lower than CEA(83 .1% and 74 .6% )(P<0 .05) .However ,the sensitivity and specificity was 90 .6% and 88 .2% when combined the two biomarkers together .Conclusion YKL‐40 have a certain clinical diagnostic value in malignant pleural effusion ,it indicate to adenocarcinoma or advanced cancer when the level of YKL‐40 rised .Since the sensitivity and specificity is lower than traditional biomarker of CEA ,we should combine with the other tumor markers to improve the diagnostic accuracy .
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[ABSTRACT]AIM:Toobservetheeffectofazithromycinontheratswithchronicobstructivepulmonarydisease ( COPD) , and to explore the underlying mechanism about the airway inflammation and mucus hypersecretion.METH-ODS:Male SD rats were randomly divided into normal control group, COPD model group, azithromycin treatment group. The COPD model was established by the method of cigarette smoking combined with intratracheal injection of LPS.Patho-logical changes of the bronchi and lung tissues of the rats were observed with HE staining.Pulmonary ventilation function in the rats was detected with pulmonary function instrument.The levels of IL-8, IL-17 and TNF-αin bronchoalveolar lavage fluid (BALF) were measured by ELISA.The expression of MUC5ac and TLR4 at mRNA and protein levels in bronchi and lung tissues was determined by real-time PCR and Western blot.RESULTS:HE staining showed that the changes of bron-chi and lung tissues in model group were consistent with typical pathological manifestations of COPD .Compared with model group, these changes were alleviated in treatment group.The pulmonary functions in model group were significantly de-creased compared with control group.The levels of IL-8, IL-17 and TNF-αin the BALF in model group were significantly increased compared with control group (P<0.05).The expression of MUC5ac and TLR4 at mRNA and protein levels in model group was significantly higher than that in control group (P<0.05).Compared with model group, the degree of the descent in pulmonary function in treatment group was significantly lessened.Compared with model group, the levels of IL-8, IL-17 and TNF-αin treatment group were significantly inhibited (P<0.05).Furthermore, the expression of MUC5ac and TLR4 at mRNA and protein levels in treatment group was significantly lower than that in model group ( P<0.05 ) . CONCLUSION:Azithromycin decreases the levels of IL-8, IL-17 and TNF-αin the BALF of COPD model rats, inhibits the protein expression of MUC5ac and TLR4 in the lung tissues, thus playing a preventive and therapeutic role to reduce airway inflammation and airway mucus hypersecretion.
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Aim To investigate the role of cAMP re-sponse element binding protein (CREB)in the injury of rat pulmonary microvascular endothelial cell (RPM-VEC)induced by LPS.Methods RPMVECs were i-solated and cultured in vitro,Western-blot was used to assay phosphorylation levels of CREB.Endothelial per-meability was determined by measuring the influx of Evans blue-labeled albumin across endothelial mono-layer.Results LPS increased CREB phosphorylation at Ser 1 3 3 in RPMVEC in a time-dependent manner , peaked at 30 min,but still higher at 120 min compared with basal control group.Pretreatment of cells with PKA inhibitor V5681 nearly suppressed the CREB phosphorylation stimulated in the presence of LPS,and the monolayer permeability of PMVEC was significantly increased. Conclusions LPS rapidly induces the phosphorylation of CREB in RPMVEC,and PKA me-diates the process.During the process of LPS-stimula-ted injury of RPMVEC,phosphorylation of CREB may play a protective role.
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Objective To investigate the role of Ezrin and its phosphorylation(p-Ezrin)in the modulation of rat pulmonary microvascular endothelial cell(PMVEC)injury induced by tumor necrosis factor-α(TNF-α)and the impact of Rac 1. Methods Cultured PMVECs of Sprague-Dawley(SD)rats were randomly divided into time-dependent injury group induced by TNF-αand intervention group in which cells were pretreated with Rac 1 inhibitor (NSC 23766).①In the time-dependent injury group, Western Blot was used to detect the expression of Ezrin and p-Ezrin after 10μg/L TNF-αstimulation for 0,0.25,0.5,1,3,6,12,24 hours.②In the intervention group,after pre-treatment with 200μmol/L NSC 23766 for 0.5 h,PMVECs were treated with 10μg/L TNF-α,and the expression of p-Ezrin was detected by Western Blot after 3 hours. Besides these groups,there were control(1% fetal bovine serum simulation),single NSC 23766 or TNF-α simulation groups. Results ① Few Ezrin expression was found in PMVEC,and TNF-α could not affect Ezrin expression. p-Ezrin protein expression(p-Ezrin/Ezrin,gray scale) of PMVECs at 0 hour after TNF-αstimulation was 0.21±0.03,and elevated at 0.25 hour(0.53±0.19),peaked at 3 hours(1.68±0.30),then it was gradually lowered,but it remained at higher level at 24 hours(0.87±0.18)with significant difference(F=62.200,P=0.000). It demonstrated that TNF-αcould increase Ezrin phosphorylation in a time-dependent manner.②Compared with blank control group,in single NSC 23766 or TNF-αsimulation group, p-Ezrin expression was induced(TNF-αgroup:0.92±0.12 vs. 0.68±0.16,t=-2.864,P=0.020;NSC 23766 group:1.33±0.24 vs. 0.68±0.16,t=-5.429,P=0.000. When NSC 23766 was pre-treated with PMVECs,the expression of p-Ezrin was significantly increased compared with that in single TNF-αsimulation group(2.14±0.18 vs. 0.92±0.12,t=-14.670,P=0.000)with significant difference(F=73.810,P=0.000). Conclusion Ezrin proteins are phosphorylated by TNF-α. Rac 1 signaling pathway inhibition plays an important role in TNF-α-induced injury by up-regulation of p-Ezrin in PMVECs.
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Objective To study the role of src-suppressed C kinase substrate (SSeCKS) in the secretion of tumor necrosis factor (TNF-α) in rat pulmonary micro-vascular endothelial cells (PMVEC) induced by lipopolysaccharide (LPS).Methods Wistar rat PMVEC cultured in vitro were randomly (random number) divided into several groups (n =4) as per exposure to given dosage of LPS for different lengths of time and to different dosages of LPS for given length of time.After PMVEC exposed to 10 mg/L LPS for 1 hour (h),3 h,6 h,12 h and 24 h or 0.1 mg/L,1 mg/L and 10 mg/L LPS for 24 h,the levels of TNF-αin the supernatant of culture medium were examined by the method of enzyme linked immunosorbent assay (ELISA).Another PMVEC was pre-treated by protein kinase C (PKC) inhibitor bis-indolylmaleimide (BIM) for 0.5 h or had the transfection of SSeCKS-specific small interfering RNA (siRNA) for 48 h before 10 mg/L LPS challenge for 24 h,and subsequently the supernatant was also examined by ELISA.One-way analysis of variance (ANOVA) was employed for statistical analysis by SPSS version 10.0 to compare values among all groups.A significant difference was presumed as a probability value ＜ 0.05.Results After PMVEC incubated with 0.1 mg/L,1 mg/L and 10 mg/L LPS for 24 hours,the levels of TNF-αsecreted were (253.70 ± 23.55),(327.88 ± 37.25),(403.20 ± 36.22),respectively,which were higher than that in un-stimulated PMVEC (82.28 ± 22.56,all P =0.000).After 10 mg/L LPS challenge for one hour,the level of TNF-αin the supernatant of PMVEC raised substantially (170.11 ±49.22),peaked at the time of 6 h (404.82 ± 13.78),then persisted at a higher level until 24 h (395.67 ± 36.23) than that in un-stimulated PMVEC (84.60 ± 23.61,P =0.001,0.000,0.000,respectively).After PMVEC pre-incubated with BIM,the level of LPS-induced TNF-αdecreased obviously (200.44 ± 27.39 vs.402.28 ± 31.07,P =0.000).Compared with LPS challenged PMVEC (407.28 ± 32.64),depletion of endogenous SSeCKS in PMVEC after inhibited by SSeCKS-siRNA significantly attenuated increase in the level of LPS-induced TNF-α (195.20 ± 13.28,P =0.000).Conclusions Down-activation of SSeCKS and PKC can inhibit the secretion of TNF-αin PMVEC induced by LPS,relieving the inflammatory response of PMVEC.
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Objective To study the role of soluble intercellular adhesion molecule-1 ( sICAM-1 ) in the pathogenesis of adults bronchial asthma and analyze the relationship between the level of sICAM-1 and the severity of bronchial asthma.Methods Serum levels of sICAM-1 in 134 cases with different periods of bronchial asthma patients and healthy volunteers were measured by ELISA and pulmonary functions of acute asthma patients were determined by pulmonary function analyzer. Results Serum sICAM-1 levels in 63 cases of acute asthma patients were increased significantly when compared with those in remission asthma patients and healthy volunteers and its value was increased significantly with the exacerbation of asthma;A negatively correlation between FEV1％ of pulmonary function and serum sICAM-1 level in patients with acute asthma was found in this study. Conclusion sICAM-1 was one of the important adhesion molecules in the pathogenesis of bronchial asthma. It could be used as one indicator of disease severity and guide the drug application in clinic.
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OBJECTIVE To study the therapeutic effect of ceftazidime(CAZ) in a rat model of pneumonia caused by CTX-M-14 extended-spectrum ?-lactamases(ESBLs)-producing Klebsiella pneumoniae which is susceptible to ceftazidime in vitro.METHODS Forty eight rats were divided into four groups randomly which were ceftazidime-treated group,piperacillin/tazobactam-treated group,ceftaxime-treated group and the control group.All rats were intranasally inoculated with bacterial suspension of CTX-M-14 ESBLs-producing K.pneumoniae to produce pneumonia models.Twenty four hours after inoculation,rats were intramuscularly administered ceftazidime,piperacillin/tazobactam,ceftaxime,or saline,respectively.The temperature,leucocyte count,percentage of neutrophils,and bacterial counts in the right lungs of rats were detected at some timepoints.RESULTS Compared with the ceftaxime-treated group and the control group,the leucocyte count,percentage of neutrophil count and common logarithm of the bacterial count in the right lungs of the ceftazidime-treated group and piperacillin/tazobactam-treated group after the treatment which lasted seventy two hours were significantly lower than those of ceftaxime-treated group and the control group(P0.05).CONCLUSIONS Ceftazidime shows good therapeutic effect in pneumonia in rats caused by CTX-M-14 ESBLs-producing K.pneumoniae which is susceptible in vitro.
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Aim To investigate the possible mechanism of antitumor in human lung cancer cell lines A549 and NCI-H460 induced by lumiracoxib.Methods The expression of COX-2 was detected by Western blot and the levels of PGE2 and cAMP was determined by radioimmunoassay (RIA).Results COX-2 protein was highly expressed in A549 and NCI-H460 cells.After treatment with 15~240 ?mol?L-1 LUM for 24 hrs,LUM significantly decreased the level of COX-2 in A549 cells,but not in NCI-H460 cells.Compared with the control,the PGE2 production was reduced and the level of cAMP was increased after the treatment with 15,30,60,120,240 ?mol?L-1 of LUM,respectively.Conclusion The effect of Lumiracoxib on antitumor is in COX-2-dependent or-independent manner. The antitumor effect of LUM may be related to inhibiting the COX-2 activities by decreasing its secretion,up-regulating the level of cAMP,and down-regulating the level of PGE2.
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Aim To establish an animal model of inflammatory pleural adhesion and to investigate the role of low-molecular-weight heparin(LMWH) in preventing encapsulation of pleural effusion.Methods Thirty New-Zealand rabbits were randomly divided into three groups:the model and the treatment group received injection of tetracycline(35 mg?kg~(-1)) in the right chest cavity,the control animals received injection of normal saline.LMWH (55 U?kg~(-1)) was injected into the right chest cavity of rabbits in the treatment group.The animals were killed on 14 d.Result In the model group,pleural effusion appeared at 2 h and reached maximum within 2～3 days.In the effusion,leukocytes accumulated and procoagulant activity increased at the same time.Fibrosis and pleural adhesion,encapsulation appeared on the fourteenth day.In the treatment group pleural effusion and procoagulant activity were reduced.No pleural adhesion was observed.Conclusion Tetracycline was successfully used to establish an animal model of pleural adhesion in rabbits.LMWH was effective in reducing pleural effusion and preventing pleural fibrosis and encapsulation.
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Objective To investigate the effects of tumor necrosis factor ? (TNF ?) on ? adrenoceptor (? AR) and ? AR related G protein coupled receptor kinases (GRKs) in rat pulmonary microvascular endothelial cells (PMVECs) as well as the interfering action of anisodamine. Methods Radio ligand binding assay was used to measure the maximal binding capacity (B max ) changes of ? AR in rat PMVECs after treatment with TNF ?. The effects of TNF ? and ? AR related GRKs expression at the protein level were observed by Western blotting. Results B max of ? AR in normal rat PMVECs was (5.58?0.31) fmol/10 5 cells. B max of ? AR in TNF ? group decreased significantly as compared with that in the normal control group, but no significant difference was found between the normal control group and TNF ?+anisodamine group. The expression of GRK2 in rat PMVECs was positive, but expression of GRK3, GRK5, and GRK6 were negative. The expression of GRK2 in TNF ? group and TNF ?+anisodamine group increased significantly as compared with that in the normal control group, but no significant difference was found between the TNF ? group and the TNF ?+anisodamine group. Conclusion GRK2 but not GRK3, GRK5, or GRK6 is expressed in rat PMVECs. The increased expression of GRK2 induced by TNF ? in rat PMVECs might promote the phosphorylation of ? AR, leading to ? AR internalization and decoupling with G protein, which might be the main mechanism of down regulation of ? AR induced by TNF ?. Anisodamine might inhibit the down regulation of ? AR through other mechanism instead of inhibiting the increase of GRK2 expression.
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Aim To investigate the mechanism of platelet-activating factor(PAF) induced injury on the cultured rat pulmonary microvascular endothelial cells(RPMVEC),and the interfering action of breviscapine. Methods RPMVEC were isolated from Wistar rat and cultured in vitro, the effects of PAF on morphology,monolayer permeability and F-actin of RPMVEC were observed, F-actin expression was evaluated by flow cytometry. Results PAF in the concentration over 0.1 mg?L -1 induced detachment and rupture of RPMVEC within 48 h. 10 mg?L -1 of PAF increased permeability of RPMVEC monolayer and induced depolymerization of F-actin within 120 min.Breviscapine inhibited these effects of PAF. Conclusion ①The detachment and rupture of RPMVEC induced by PAF depends on the exposed concentration and time.②The increased permeability of RPMVEC monolayer induced by PAF is significantly correlated with the depolymerization of F-actin. ③The increased permeability of RPMVEC monolayer and depolymerization of F-actin induced by PAF can be markedly inhibited by breviscapine, which exerts protective action on RPMVEC injury induced by PAF.
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G protein-coupleed receptor kinases (GRKs)not only regulates phosphorylation of G protein-coupled receptor which mediates receptor desensitization and initiates profound impairment of receptor signaling,but also regulates G protein and cytoskeleton. Meanwhile GRKs are regulated by protein kinase A, protein kinase C,actin and calcium/calmodulin. Surface of cell has many kinds of G protein-coupled receptor such as PAF receptor, histamine receptor, thrombine receptor which initiates signal effect of cell injury induced by inflammatory mediator. So GRKs have a certain regulatory role in the process of inflammation-induced cell injury through phosphorates G protein-coupled receptor.
ABSTRACT
Aim To study the expression of protein kinase C(PKC) subtype PKC? in rat pulmonary microvascular endothelial cell(RPMVEC),and the effect of its inhibitor(Rottlerin) on LPS-induced RPMVEC monolayer permeability injury.Methods In cultured RPMVEC,Western blot and immunohistochemistry were used to identify the expression of PKC?.Micro-infiltrator was used to measure the changes of RPMVEC monolayer permeability coefficient(Kf) after exposure to LPS or LPS with Rotterin together.Result We found a high level of PKC? expression in cytoplasm.Rottlerin could obviously inhibit the increase of LPS-induced RPMVEC Kf value.Conclusion The activation of PKC is involved in the progress of LPS-induced RPMVEC injury.Furthermore,the inhibitor of PKC? could release LPS-induced RPMVEC Kf increase.
ABSTRACT
AIM Observing the influence of lipopolysaccharide (LPS) on changes of the permeability and actin cytoskeleton of rat pulmonary microvascular endothelial cells (PMVEC) monolayer, we explore their relationship with β-adrenoceptor(β-AR) and G protein subunit(Gsα),and the interfering action of anisodamine. METHODS PMVEC was isolated and cultured from Wistar rat in vitro, radioligand binding assay and flow cytometer were used. RESULTS After incubation of LPS in vitro, the central F-actin of PMVEC depolymerized and its density decreased obviously ,while the permeability of PMVEC monolyer increased significantly. Meanwhile it was found that LPS can induce the down regulation of β-AR and Gsα protein level. Anisodamine can inhibit these changes above. CONCLUSION LPS can directly cause the increase of permeability of PMVEC monolayer which relates to the depolymerization of central F-actin in PMVEC. Anisodamine may take part in regulating actin cytoskeleton of PMVEC and attenuate LPS-induced the increase of permeability of PMVEC monolayer by influencing two links of both β-AR and Gsα protein.