Objective To investigate the effects of heme oxygenase-1 (HO-1) on the signal transduction pathway of mucin(MUC) expression induced by cigarette smoke.Methods The cell model of mucous hypersecretion was made by human lung A549 cell stimulated by cigarette smoke extract(CSE),the cells were divided into 4 groups:negative control group,CSE treatment group,heroin pre-treatment group and ZnPPIX pre-treatment group.The expression of MUC5AC,epidermal growth factor receptor(EGFR),p-EGFR,HO-1 and dual oxidase 1 (Duoxl) were detected.The cell activity was assessed by methyl thiazolyl tetrazolium method.The change of HO-1 mRNA,Duoxl mRNA,EGFR mRNA and MUC5AC mRNA was examined by reverse transcriptase-polymerase chain reaction.The protein expression of EGFR,p-EGFR,Duox1 and HO-1 was measured by Western blot,while the pro tein expression changes of MUC 5 AC were detected by ELISA.Results The expression level of MUC 5 AC mRNA and its protein in the CSE group increased significantly (P＜0.05) as compared to those in the control group [0.412±0.043,(105±8) μg/mg.The mRNA and protein of EGFR,Duox1 and HO-1,the protein of p-EGFR increased significantly as compared to the control group.After the cells being pre-treated with hemin,the mRNA and protein of HO-1 increased significantly,while the mRNA and protein of Duox1,EGFR and MUC,SAC,the protein of p-EGFR decreased significantly as compared to the CSE group.After the cells were pre-treated with ZnPPIX,the increase of HO-1 mRNA and protein was not significant as compared to the control group,while the mRNA and protein of Duox1,EGFR and MUCSAC,the protein of p-EGFR increased significantly.Conclusion HO-1 decreased the level of Duox1,blocked ligand-dependent EGF-R activation and decreased the expression levels of MUCSAC.

Objective To explore the mutual effect of proinflammatory factor tumor necrosis factor (TNF)-α and cigarette smoke extract(CSE) in the induction of mucin 5AC in BEAS-2B human airway epithelial cells. Methods The BEAS-2B cells were transfected with nuclear factor(NF)-κB decoy oligonucleotides(ODNs) or transfected with NF-κB scrambled ODNs as negative control,then treated with TNF-α,CSE,and TNF-α plus CSE, respectively. The expression of phosphorylated epidermal growth factor receptor (p-EGFR) , mucin 5AC( MUC5AC) protein produc-tion and gene expression were detected by Western blot, ELISA and RT-PCR respectively. Results Significant in-creasing of p-EGFR protein production in scrambled ODNs transfected cells with elevation of MUC5AC protein production and mRNA expression was recorded and TNF-α and CSE co-incubated groups were much higher than single TNF-α or CSE stimulated group(P <0. 05). In NF-κB decoy ODNs transfected cells,TNF-α stimuli revealed an obvious reduction of MUC5AC protein production and gene expression. Conclusion TNF-α and CSE can syner-gistically induce mucin 5AC expression in BEAS-2B cells, transcription factor NF-κB plays a critical role in TNF-α induced mucin production but not as so important in mucin production by CSE stimulated cells.

Objective This single-center prospective,case-controlled study was carried out to investigate clinical effects on patients with aging face who underwent facial autologous fat grafting with platelet-rich plasma (PRP).Methods Thirty patients with facial sagging or partial depression and requiring autologous fat grafting were randomly selected for the study.Photograph and 3D scanning were taken before and 3-month after the operation.Patient and physician satisfaction was rated using Visual Analogue Scale (VAS) ranging from 1 (most unsatisfactory) to 10 (most satisfactory)3 months after the operation.Statistical difference between the patient and physician satisfactory scores was analyzed.Results The average physician VAS score was 7.9 ± 1.0 while the average patient VAS score was 8.0±1.2.The scores between two groups had no significant difference (P＞ 0.05).Four patients developed bruising and swelling postoperatively that disappeared less than 7 days later.There was no feed back of serious complications from patients.Further investigation of patients with scores less than 7 showed the main reason of low patient satisfaction was that the improvement of facial depression did not meet the requirements of the patient,and fat absorption led to unsatisfactory filling.Conclusions The high satisfactory scores indicate good aesthetic result of facial autologous fat grafting with PRP.PRP may lead to higer survival rate of fat and satisfaction.

Objective To explore the upstream signal transduction pathway of neutrophil elastase(NE)-induced mucin(MUC)5AC gene expression.Methods A549 cells were either incubated with NE alone or with DMTU,aprotinin or AG1478.The level of MUC5AC mRNA was measured with RT-PCR.The activation of epidermal growth factor receptor(EGFR) and signaling were assessed by measuring the release of epidermal growth factor(EGF) and the phosphorylation of EGFR.Results NE increased MUC5AC gene expression accompanied by an increase of EGF and phosphorylated EGFR.DMTU,aprotinin and AG1478 significantly inhibited MUC5AC gene expression.DMTU and aprotinin significantly decreased the level of EGF and phosphorylated EGFR.ConclusionNE can induce MUC5AC gene expression via EGFR signalling in A549 cells.The upper stream involves oxidants,activation of tissue kallikrein and EGF.

OBJECTIVE To observe whether the natural antibacterial polypeptide elafin after transfected into airway epithelial cells(A549 cells)has resistant effects on Pseudomonas aeruginosa bioflim.METHODS To establish the P.aeruginosa biofilm model in vitro,a rapid silver nitrate staining procedure was used to demonstrate bacterial biofilm.After cultivating the cells in vitro,the constructed pEGF-N1-elafin eukaryotic expression vector had transfected into A549 cells by LipofectamineTM 2000.Elafin transfected cells were incubated by pig Pancreas Elastase(NE group),the supernatants of PAE(PAE group)and Escherichia coli(ECO group),respectively,for 24 hours.Then the content of elafin in cells and the level of elafin secretion were detected by Western blot and ELISA respectively.After biofilm carriers were put into each group and incubated for 8 hours,we observed the ratio of cells shape breakage at 4,8,12 and 24 hour,respectively.RESULTS The NE,PAE and ECO groups induced the content of elafin in cells and the level of secretion increased compared to the no induced group(normal group),while the PAE group and NE group were higher than those of ECO group.The ratio of cells breakage in induced elafin groups was lower than that of in normal group(P

Objective To explore the molecule mechanism of protective effects of transfecting human elafin gene recombinant plasmid to NCI-H292.Methods Constructe eukaryotic expression vector pEGFP-C1-elafin and transfectied it into NCI-H292 cells,then co-incubated with polymorphonuclear granulocyte(PMN) with stimulation by lipopolysaccharide(LPS).Afater 24 hour,Western bolt and MTT technique were respectively performed to detect expression of ZO-1 in cells and adhesion capability of cells with collagen.Results After stimulation with LPS,Western blot showed that the 220 ku onula occludens-1(ZO-1) protein in transfecting pEGFP-C1 NCI-H292 cells extracts were obviously decreased,in transfecting pEGFP-C1-elafin NCI-H292 cells extracts.And MTT technique showed that compared with transfecting pEGFP-C1-elafin cells,adhesion capability of transfecting pEGFP-C1 cells with collagen were obviously decreased.Conclusion Transfection of recombinant human elafin gene into airway epithelial cells could maintain airway epithelium and enhance the capability of airway to inflammatory injury.

Objective To explore the mutual effect of proinflammatory factor tumor necrosis factor(TNF)-? and cigarette smoke extract(CSE) in the induction of mucin 5AC in BEAS-2B human airway epithelial cells.MethodsThe BEAS-2B cells were transfected with nuclear factor(NF)-?B decoy oligonucleotides(ODNs) or transfected with NF-?B scrambled ODNs as negative control,then treated with TNF-?,CSE,and TNF-? plus CSE,respectively. The expression of phosphorylated epidermal growth factor receptor(p-EGFR),mucin 5AC(MUC5AC) protein production and gene expression were detected by Western blot,ELISA and RT-PCR respectively.Results Significant increasing of p-EGFR protein production in scrambled ODNs transfected cells with elevation of MUC5AC protein production and mRNA expression was recorded and TNF-? and CSE co-incubated groups were much higher than single TNF-? or CSE stimulated group(P

The method for examining the microcirculation bloodflow of mice mesentery was used to evaluate the action of Shujinhuoxuedingtong wine against microcirculation disturbance caused by adrenaline. The result indicates that the wine can obviously improve the microcirculation function and is responsible for the antistatic action observed. The wine's effect is found to be superior to that of Shujinhuoxuedingtong Powder. The above fingings suggest that the wine have the action of promoting blood circulation to remove blood stasis and improving microcirculation and the effect of promoting the bloodflow perfusion quantity in tissue organs. It's effect is prior to that of powder.

Objective To construct the recombinant adenovirus for nature antibiotic elafin protein and observe its expression in primary airway epithelia.Methods The elafin gene was amplified by RT-PCR from lung tissues,then the fragment was inserted into the multiple clone site of pAdTrack-CMV.The linearized shuttle plasmid was homogenously recombined with AdEasy-1 in BJ5183.The candidate clone was further analyzed by restriction endonuclease digestion,PCR,and sequencing.Then the recombinant adenovirus plasmid was digested with PacⅠ and transfected into 293 cells for packaging and amplification.Infection titer and rate were monitored by green fluorescent protein(GFP)expression.Finally the primary airway epithelium was infected with Ad-elafin,and the GFP expression was observed by fluorescence microscope in epithelial cells 24 h after transfection.The elafin protein was detected by ELISA in the supernatant.Results Restriction endonuclease and PCR confirmed that elafin gene was cloned into the adenovirus vector successfully.Twenty-four hours after transfection,the GFP expression was seen by fluorescence microscopy.The concentration of elafin protein was(2.2?0.4)ng/ml in supernatant of transfected group,while that of control group was(1.9?0.3)ng/ml,P

Objective To synthesize a marine natural product, 24-methylenecholesterol. Methods The target molecule was synthesized in 7 steps using hyodeoxycholic acid as the starting material, 3?-(tetrahydropyran-2-yloxy)-cholest-5-ene-24-ketone as the key intermediate and isopropylation as the key step. Results The target molecule was synthesized with a total yield of 66% and confirmed by MS, IR, 1HNMR and 13CNMR. Conclusion Our synthesis of 24-methylenecholesterol is characterized by easily acquired starting material, short synthetic route, high yield and atom-economic.