Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Female ; Fluorouracil ; pharmacology ; Gene Expression Regulation, Neoplastic ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; physiology ; Neoplasm Proteins ; biosynthesis ; genetics ; physiology ; Oligodeoxyribonucleotides, Antisense ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transfection

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.

OBJECTIVETo study the distribution and antibiotic resistance of the isolated pathogens from children with infectious diarrhea in Guangzhou.

METHODSThe fecal samples of 2 409 children with infectious diarrhea between January 2006 and December 2007 were collected and cultured. Pathogenic bacterium were isolated and identified by biochemical and serological methods. The antibiotic susceptibilities were tested by the Kirby-Bauer method.

RESULTSA total of 448 isolates of pathogenic bacterium (18.6%) were obtained, including Shigella (n=159), enteropathogenic Escherichia coli (n=141), Salmonella (n=76), Vibrion (n=11), fungus (n=41), and C jejuni (n=20). All of isolates of the three major pathogenic bacterium, Shigella, enteropathogenic Escherichia coli and Salmonella, were susceptible to imipenem and less than 10% of the isolates were resistant to the third generation cephalosporins and beta-lactamase inhibitors. However, the isolates showed a high resistance to ampicillin and sulfamethoxazole/trimethoprim (>75%).

CONCLUSIONSShigella, enteropathogenic Escherichia coli and Salmonella were major pathogenic bacterium of diarrhea in children from Guangzhou. The major isolates were susceptible to imipenem, the third generation cephalosporins and beta -lactamase inhibitors, but were resistant to ampicillin and sulfamethoxazole/trimethoprim.

Adolescent ; Bacteria ; drug effects ; isolation & purification ; Child ; Child, Preschool ; Diarrhea ; drug therapy ; microbiology ; Drug Resistance, Microbial ; Female ; Fungi ; drug effects ; isolation & purification ; Humans ; Infant ; Male

There are similarities between magnetotactic bacteria and Acidithiobacillus ferrooxidans (A. ferrooxidans) which isolated from Acid mine drainage(AMD). The weak magnetotaxis of some bioleaching bacteria isolated were found by microscope. A magnetophoresis apparatus was designed based on these weak magnetotaxis and be used to analysis the movement of these strains. The physiological properties of the anear magnetic field strain and removed magnetic field strain which isolated successfully by magnetophoresis apparatus have large difference. The nanometer magnetic particles was extract from the Acidithiobacillus ferrooxidans which purified by spread plate method from AMFS and its main elements are Fe and O by energy spectrum analysis. The results show that A. ferrooxidans have weak magnetotaxis and can be isolated by magnetophoresis. With the development of this new isolating method, the research of magnetotactic bacteria and bioleaching will get more benefit from it.

Objective To observe the effect of sinomenine (SIN) on the expression of cyclooxygenase (COX2),alpha-7 nicotinic acetylcholine receptor(α7nAChR) and adenosine receptor(A2A) in A549 cells,and to explore the relative mechanism for cell proliferation.Methods The effect of SIN and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) on the proliferation of A549 cells was determined by methyl thiazolyl tetrazolium (MTT) assay.The effect of SIN and NNK on the migration of A549 cells was detected by cell wound scratch assay.The effect of SIN and NNK on COX2 expression in A549 cells was determined by Western blotting method.The effect of SIN and NNK on the expression of α7nAChR and A2A mRNA and protein was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting method.Results NNK increased the proliferation and migration of A549 cells,while SIN inhibited the proliferation and migration of A549 cells.COX2 expression level was increased in NNK group but was decreased in SIN group.The expression levels of α7nAChR and A2A were up-regulated in NNK group but were down-regulated in SIN group.Conclusion SIN plays a role in inhibiting the proliferation and migration of A549 cells by suppressing COX2 expression.SIN has an inhibitory effect on the expression of α7nAChR and A2A.

AIM:To evaluate 23G vitrectomy for macular edema in eyes with retinal vein occlusion (RVO) combined with vitreoretinal traction (VMT) or epiretinal membrane (ERM).METHODS:Totally 22 patients (22 eyes) diagnosed with macular edema of RVO combined with VMT or ERM were retrospectively analyzed.Twelve cases performed with 23G vitrectomy together with peeling of inner limiting membrane (ILM) and/or ERM were considered as the observation group or intervention group.Ten cases without vitrectomy were recruited as control group.The best corrected visual acuity (BCVA) and central retinal thickness (CRT) at baseline, 1, 3 and 6mo were recorded and compared.RESULTS:At baseline, the difference of BCVA and CRT between observation group and control group was not statistically significant (P=0.645, 0.206).After vitrectomy, the BCVA and CRT of RVO patients in observation group were significantly improved compared with baseline at each follow-up (F=2.895, P=0.048;F=16.431, P<0.01).However, the BCVA and CRT in control group remained the same as baseline at every follow-up.Moreover, the BCVA and CRT in observation group were much better than that in control group at both 3 and 6mo after vitrectomy.However, the BCVA and CRT between two groups were not significantly different at 1mo postoperatively.CONCLUSION:The 23G vitrectomy could markedly improve BCVA and reduce CRT in RVO patients with macular edema combined with VMT and/or ERM.

Acute Kidney Injury ; therapy ; Heart Defects, Congenital ; surgery ; Humans ; Infant, Newborn ; Male ; Peritoneal Dialysis ; Postoperative Complications ; etiology

OBJECTIVETo establish a method for rapid differential diagnosis of thalassemia trait (TT) and iron-deficiency anemia (IDA) using stepwise regression analysis.

METHODSStepwise regression equation was established for differential diagnosis of TT and IDA according to the red cell index, and the accuracy of the differential diagnosis was evaluated using blind analysis.

RESULTSThe accuracy of this equation for differential diagnosis of TT and IDA was 86.82%. The sensitivity, specificity and Youden's index in prediction of TT and IDA were 94.29%, 79.66%, 73.9 and 76.92%, 90.52%, and 67.4%, respectively.

CONCLUSIONThe stepwise regression equation using the red cell index is concise, rapid, and sensitive in differential diagnosis of TT and IDA, and can be well applicable in clinical practice.

Adult ; Anemia, Iron-Deficiency ; blood ; diagnosis ; Diagnosis, Differential ; Double-Blind Method ; Female ; Humans ; Male ; Regression Analysis ; Reproducibility of Results ; Sensitivity and Specificity ; Thalassemia ; blood ; diagnosis

OBJECTIVETo study the influence of human cytomegalovirus (HCMV) infection on cell cycle and the expression of replication licensing factor Cdt1 in human embryonic lung fibroblastic (HEL) cells and to explore the pathogenesis of HCMV infection.

METHODSHEL cells were synchronized in the G0/G1 phase by the serum starvation method. The synchronized HEL cells were infected with HCMV, and those that were not subjected to HCMV infection were used as the control group. The HEL cells were harvested at 12, 24, 48, 72 and 96 hrs of HCMV infection. The cell cycle of HEL cells was detected by the flow cytometry. The expression of Cdt1 mRNA in HEL cells was determined by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe cells in the G1 phase in the control group was significantly more than in the HCMV-infected group 12 and 24 hrs after infection (P < 0.01). The expression of Cdt1 mRNA in the HCMV-infected group was significantly lower 12 and 24 hrs after infection but increased significantly 48 hrs after infection compared with the control group (P < 0.05). The expression of Cdt1 mRNA reached a peak at 12 hrs of infection in the control group, but at 48 hrs of infection in the HCMV-infected group, which markedly lagged behind the control group.

CONCLUSIONSHCMV infection arrests the cell cycle of HEL cells at the G1 phase. HCMV infection makes Cdt1 expression delay. HCMV infection can interfere cell cycle of HEL cells possibly through affecting the expression of Cdt1.

Cell Cycle ; Cell Cycle Proteins ; genetics ; Cells, Cultured ; Cytomegalovirus ; pathogenicity ; Embryo, Mammalian ; cytology ; Fibroblasts ; cytology ; metabolism ; Humans ; Lung ; cytology ; metabolism ; RNA, Messenger ; analysis

In order to analyze the molecular epidemiology of Hantavirus (HV) in Zhejiang Province, the complete M and S genome sequences of 12 HV strains from different hosts and locations in Zhejiang Province of China during the period of 1981-2007 were analyzed on genetic evolution by DNAstar and MEGA 4.0 software in this research. Phylogenetic analyses revealed that HTN and SEO strains were co-circulating in Zhejiang Province, and the difference in sequence similarity and the phylogeny was closely related to the isolated regions, but had no distinct relationship with the isolate year and the host, indicating a relationship between epidemiology of HFRS and the distribution region, especially in HTNV. The isolates in the same region could be assigned in same or near phylogenetic clade sharing high sequence similarity. Interestingly, the Gou3 strain and ZJ5 strain isolated from Jiande region in Zhejiang Province formed a distinct phylogenetic lineage in SEOV clade, and different from the other SEOV variants outside China. We believed that the special SEOV variants were distributed in Jiande region.
Animals ; China ; Disease Reservoirs ; virology ; Evolution, Molecular ; Hantavirus ; classification ; genetics ; isolation & purification ; Hantavirus Infections ; virology ; Humans ; Molecular Sequence Data ; Phylogeny ; Rodentia ; virology ; Viral Proteins ; genetics