Objective To investigate the killing effect of ethacrynic acid (EA) on lung cancer A549 cells derived spheres and explore the underlying mechanism.Methods A549 spheres were cultured in serum-free medium,and the protein expression of CD133,SOX2,EpCAM and ABCG2 was detected by Western blotting.MTT assay was used to evaluate the cell viability of A549 spheres and A549 cells after treated by 1,2,5,10 and 20 mg/mL cisplatin (DDP) for 48 h.The activity of glutathione S-transferase (GST) was measured by colorimetric method after A549 spheres were treated with 10,50,100 and 200 μmol/L EA,respectively.Flow cytometry,Western blotting,real-time PCR and luciferase assay were used to analyze the levels of cellular reactive oxygen species (ROS),formation of A549 spheres,mRNA and protein expression levels of β-catenin,Sox2 and ABCG2,and promoter activity of β-catenin upon 200 μmol/L EA treated cells for 48 h.A549 sphere was infected with β-catenin adenovirus for 24 h,followed by 200 μmol/L EA treatment (in presence or absence of 5 mg/mL DDP) for 24 h.The expression of β-catenin,Sox2 and ABCG2 at mRNA and protein levels was detected by real-time PCR and Western blotting,and cell growth of A549 spheres was evaluated by MTT assay.Results The A549 spheres,with high expression of tumor stem cells markers CD133,SOX2,EpCAM and drug resistance related molecule ABCG2,and resistance to DDP at different doses,were successfully derived.After 200 μmol/L EA had treated A549 sphere for 48 h,the levels of ROS were significantly increased (P ＜ 0.05),and the mRNA and protein levels of β-catenin,Sox2 and ABCG2,and promoter activity of β-catenin were notably decreased (P ＜ 0.05).The treatment of 200 μmol/L EA enhanced the inhibitory effect on proliferation and the promoting effect on apoptosis in A549 spheres induced by 5 mg/mL DDP (P ＜ 0.05).Up-regulation of β-catenin by adenoviral infection partly reversed the effects of 200 μmol/L EA on suppressing the expression levels of β-catenin,Sox2 and ABCG2,compared to the spheres infected with blank adenovirus.Additionally,β-catenin over-expression significantly remitted the inhibitory effect of 200 μmol/L EA and 5 mg/mL DDP on the proliferation in A549 spheres.Conclusion EA exerts inhibitory effect on the proliferation and stemness of A549 spheres through suppressing GST activity and β-catenin expression,and then promotes cell apoptosis.EA might be a novel drug in treatment of lung cancer and cancer stem cells.

Abnormalities, Multiple ; blood ; diagnosis ; metabolism ; physiopathology ; Child ; Diagnosis, Differential ; Dwarfism ; blood ; diagnosis ; metabolism ; physiopathology ; Estradiol ; blood ; Female ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Silver-Russell Syndrome ; blood ; diagnosis ; metabolism ; physiopathology

Humans ; Infant, Newborn ; Infant, Premature ; Infant, Premature, Diseases ; microbiology ; prevention & control ; Mycoses ; prevention & control

Acute Disease ; Adult ; Female ; Humans ; Male ; Middle Aged ; Occupational Diseases ; chemically induced ; Trimethyltin Compounds ; poisoning

Air ; Chromatography, Gas ; Solvents ; Workplace

Background Granulocyte colony-stimulating factor(G-CSF)has been reported to have beneficial effect on cardiac dysfunction in post infarction and doxorubicin-induced cardiomyopathy.Objective To investigate the effects of G-CSF on cardiac remodeling in cardiac hypertrophy induced by angiotensin Ⅱ(Ang Ⅱ).Methods Thirty-six male wild type mice(WT)were allocated randomly to receive subcutaneously G-CSF(10 ?g/kg per day, n=9),or Ang Ⅱ(2.88 mg/kg per day,n=9),or Ang Ⅱ plus G-CSF(Ang Ⅱ 2.88 mg/kg+G-CSF 10 ?g/kg,n =9)for 4 weeks with untreated WT(n=9)as controls.Blood pressure and cardiac function were measured. Heart weight/body weight ratio,myocyte cross-sectional area and fibrosis area were determined.The mRNA ex- pression of osteopontin(OPN)in myocardium was detected by RT-PCR.The expressions of angiotensin converting enzyme(ACE),ACE2 and phosph-p70S6 kinase protein in myocardium were assessed by Western-Blot.Results Ang Ⅱ significantly elevated blood pressure(SBP,Ang Ⅱ:139.7?1.6 vs WT:108.7?2.3 mmHg,P0.05),but significantly attenuated the myocyte cross-sectional area(Ang Ⅱ+G-CSF:181.06?0.11 vs Ang Ⅱ:202.02?0.16 ?m~2,P

Objective To explore the clinical and pathologic characteristic of deep fungus infection under mechanical ventilation in neonate. Method The clinical datas of 18 neonates under mechanical ventilation approved with deep fungus infection by autopsy from 1985 to 2003 in our department were collected and analyzed.Results In 18 infants who survived deep fungus infection,2 cases were premature and 16 cases were mature,the birth weight and the numbers of patients were:2500 g 2 cases;Time under mechanical ventilation was 8-86 days and the average time under mechanical ventilation was 13 days. The pathogen caused deep fungus infection was Aspergillus(8/18),Candida albicans(6/18) and Mucor(4/18),respectively. The infected organ were lung(12/18),brain(2/18),liver(4/18) and gastrointestinal tract(2/18),respectively.Two of them were found been infected in lung and liver at the same time.Conclusions We must attach importance to deep fungus infection in neonate under machine ventilation.Aspergillus and Candida albicans were the main pathogen bacteria and lung was the main infected organ of neonatal deep fungus infection.Early diagnosis may be the efficiency approach to increase livability.

Objective To explore the characteristics and method that used modified septoplasty in high posi- tion debiation drawing(or)accompanying dislocation of nosal septum.Methods By performing modified septoplasty on 59 patients who had high position deviation and former dislocation of nosal septum,improvement on method was carried out,and aspects of 59 patients in operation such as data,operation method,follow-up analysis were checked. Results The follow-up visiting in the following 5 years showed that the 59 patients' nasal septum were in the cen- tre of the nose.and there was no dislocation again or no perforation.Conclusion The improved method is simple and convenient,so it can be easily used in clinics.

Objective To investigate the characteristic of ?-amyloid precursor protein (A?) processing in activated platelet in AD.Methods Thirty-six sporadic AD patients and 30 control subjects were included in this study.Blood was collected from the subjects to separate platelets.After treated by thrombin,the soluble amyloid precursor protein (APP) level in the snpernatants of platelets from 36 were analyzed by means of western blot with a specific antibody recognizing soluble APP.Meanwhile A? level was measured by radioimmunoassay.Results After treated with thrombin,the level of soluble APP in the supernatants of platelets in patients with AD decreased by 31.0% (P