Changes of pulmonary ?_1-and ?-adrenergic receptor (?_1-AR and ?-AR)during endotoxin-induced acute lung injury in rates were oberved to find out the rela-tionship between them and the mechanism of change. Results showed that there was amarked decrease of B_max of both ?_1-AR and ?-AR by 35% and 43% respectively, dur-ing acute lung injury. The down regulation of ?-AR might be one of causes of acutelung injury while that of ?_1-AR seems to be a protective response. Active oxygen playedan important role in endotoxin-induced down regulation of AR in the rat lungs. The increa-sed level of norepinephrine and epinephrine was not the main factor that initiate the downregulation of AR. Intravenous injection of tumor necrosis factor (5?10~6U/kg ) exerts noiafluence on the changes of pulmonary AR in rats.

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.

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.

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.

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.

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.

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(P< 0 .05) .The elevated level of YKL‐40 was related to tumor types and lymphatic metastasis(P<0 .05) ,but it had no correlation with the gender ,age and degree of tumor differentiation(P>0 .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 .

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.

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.

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.