In order to investigate the effect of Polo-like kinase-1 (Plk1) depletion on cell cycle progression and cell growth in lung cancer cells, a recombinant plasmid containing antisense RNA targeting Plk1 (pcDNA3-Plk1) was transfected into A549 cells by lipofectine. RT-PCR and Western-blot were used to detect the Plk1 gene expression. Cell proliferation was evaluated by direct cell counting and bromodeoxyuridine (BrdU) labeling. Cell cycle distribution and apoptosis were examined by flow cytometry, and the inhibition rate (IR) by vinorebline (NVB) was determined by MTT assay. The results showed that after transfection of pcDNA3-Plk1 into A549 cells, the expression levels of Plk1 mRNA and protein were greatly decreased. In pcDNA3-Plk1 transfected groups, abnormal morphological changes of cells and growth inhibition were observed, and the BrdU labeling index was significantly lower than in the control groups (P<0.05). Cells in pcDNA3-Plk1 transfected groups were arresed in G2/M phase and apoptosis was detectable 72 h post transfection. IR induced by vinorebline in pcDNA3-Plk1 transfected groups was significantly higher than in other groups. These data suggested that antisense RNA targeting Plk1 could suppress the Plk1 expression, and therefore, significantly inhibit cell proliferation and induce cell cycle arrest and apoptosis. Moreover, it sensitized lung cancer cells to chemotherapy.

Objective To investigate the tellurite resistance level,the presence of tellurite resistance (ter) gene cluster and their relationships in non-O157 Shiga toxin-producing Escherichia coli(STEC) isolates.Methods Tellurite resistance level was evaluated by plate dilution method and the ter gene cluster was tested by PCR.Results Only 5 of 39 non-O157 STEC isolates tested in this study were identified to have ter gene cluster,which showed relatively high levels of tellurite resistance ranging from 128 μg/ml to 512 μg/ml.In contrast,the other 34 isolates without ter gene cluster were sensitive to potassium tellurite and showed very low levels of tellurite resistance,the minimal inhibitory concentration (MIC) was ＜1 μg/ml for 29 isolates,8 μg/ml for 2 isolates and 2 μg/ml for 3 isolates.Conclusion Most non-O157 STEC isolates were sensitive to potassium tellurite.It could be concluded that much attention should be paid when screening the non-O157 STEC isolates using the selective medium supplemented with potassium tellurite.

A high efficiency phenol-degrading bacterial strain PS1 was isolated from the drainage ditch of chemical laboratory of East China Institute of Technology.PS1 is a coccus,Gram negative and can live on phenol as its sole carbon and energy source.PS1 to identified as a strain of Raoultella sp.by 16S rRNA gene sequence analysis,which can degrade and tolerate more than 3500mg/L phenol.When phenol concentration is 500mg/L and 1000mg/L,PS1 can completely degrade it in 22 h and 32h,respectively.And while it is between 1500mg/L～3000mg/L,all phenol can be degraded by PS1 in 32h～50h.When phenol concentration is 2500mg/L,the phenol-degrading rate is the biggest and can reach to 78.1mg/h.The optimum growth and phenol-degrading conditions were obtained by orthogonal experiment,which are 25℃,pH6.5,glucose concentration 500mg/L and 20℃,pH7.0,glucose concentration 500mg/L,respectively.

AIM:To investigate the effect of Polo-like kinase-1(Plk1) depletion on cell cycle progression and cell growth in lung cancer cells.METHODS:A recombinant plasmid containing antisense RNA targeting Plk1(pcDNA3-Plk1) was transfected into A549 cells by lipofectine.RT-PCR and Western blotting were used to examine Plk1 gene expression.Cell proliferation was evaluated by cell counting and BrdU labeling.Cell cycle distribution and apoptosis were examined by flow cytometry.Inhibition rate(IR) of vinorebline(NVB) was determined by MTT assay.RESULTS:After transfected with pcDNA3-Plk1 into A549 cells,the expression levels of Plk1 mRNA and protein were greatly decreased.Abnormal morphological changes of cells and growth inhibition were observed in pcDNA3-Plk1 transfected cells.The BrdU labeling index was significantly lower than that in control group(P

AIM: To detect the changes of serum vascular endothelial growth factor(VEGF) level and the expression of peripheral blood cytokeratin 19(CK19) mRNA in patients with non-small cell lung cancer(NSCLC).METHODS: 96 patients with NSCLC,40 patients with benign lung diseases and 25 healthy controls were investigated.The VEGF level in serum was detected by ELISA and CK19 mRNA in peripheral blood was determined by using reverse transcriptase-polymerase chain reaction(RT-PCR).RESULTS: In NSCLC group,the serum VEGF level and the positive rate of CK19 mRNA in peripheral blood were(346.3?95.6) ng/L and 63.5%,which were significantly higher than those in other two groups respectively(P0.05).VEGF serum level in the patients who showed positive CK19 mRNA in peripheral blood was(407.4?121.2) ng/L.It is significantly higher than that in the negative patients(P

Acute Disease ; Adult ; Autopsy ; Bridged-Ring Compounds ; poisoning ; Child ; Humans ; Male ; Poisoning ; pathology ; Rodenticides ; poisoning

OBJECTIVETo investigate the relationship between Mutans streptococcus (MS), Lactobacilli (LB), pH value and buffer capacity of saliva and early childhood caries (ECC).

METHODSA total of 178 children aged from 42 to 54 months were recruited from 14 urban kindergartens in Beijing. The ECC group contained 87 children with more than 5 decayed teeth, and the control group was composed of 91 caries-free children. Unstimulated (UWS, 2 ml) and stimulated (SWS, 2 ml) whole saliva were collected in each subject. The pH value and buffer capacity of saliva were measured using an electro-acidimeter (+/- 0.01pH).

RESULTSMS and LB were isolated from 96.6% and 79.3% of children with ECC, which were significantly higher than those (63.7%, 27.5%) of caries-free children (P < 0.05) respectively. The counts of MS and LB in children with ECC were approximately 10 times higher than that in caries-free children. Initial pH value and buffer capacity of SWS were significantly higher than that of UWS (P < 0.001) in both groups. The pH value and buffer capacity of both UWS and SWS in ECC children were significantly higher than caries-free children (P < 0.05). There were no significant correlations between MS, LB and pH value and buffer capacity of saliva in caries-free children. Significant correlation (r = 0.249, P < 0.05) was found between the numbers of MS and buffer capacity of stimulated saliva in ECC children.

CONCLUSIONSMS and LB were important pathogens for ECC. Lower initial pH value and buffer capacity of saliva may be an important factor of ECC.

Buffers ; Child, Preschool ; Dental Caries ; metabolism ; microbiology ; Female ; Humans ; Hydrogen-Ion Concentration ; Lactobacillus ; isolation & purification ; Male ; Saliva ; chemistry ; microbiology ; Streptococcus mutans ; isolation & purification

Objective To explore the effect and mechanism of TGF beta1/smad3 signaling pathways on apoptosis in mouse pulmonary fibrosis.Methods Fifty-four healthy male C57BL/6 mice were randomly divided into three groups:normal control (n=18),pulmonary fibrosis model (n =18) and TGF-β1/smad3 inhibitor group (n=18).Six mice in each group were randomly killed on days 7,14 and 28.Hematoxyli~eosin and Masson staining were adopted to evaluate the severity of pulmonary inflammation and fibrosis.The content of hydroxyproline (Hyp) in the lung tissues was detected by alkaline hydrolysis technique.The apoptosis was observed by tunnel apoptosis assay kit.P-smad3 and caspase3 protein expressions were assessed via Western blot.Results Lung in model mice versus normal control showed alveolar inflammatory change in 7 days and significant pulmonary fibrosis in 28 days(P＜0.05).Meanwhile,apoptosis index,hydroxyproline content,caspase3,and phosphorylated Smad3 were obviously higher in model mice than in control group (P ＜ 0.05).Compared with model group,TGF-β1/smad3 inhibitor group showed that alveolitis and pulmonary fibrosis degree,hydroxyproline content,cell apoptosis index,the expressions of p-smad3 and caspase3 were decreased at same time point (P ＜ 0.05).Conclusions TGF beta1/smad3 signaling pathways may participate the abnormal apoptosis during the development of pulmonary fibrosis,and TGF-β1/smad3 inhibitor SB431542 could inhibit this process.

Applications of network pharmacology are increasingly widespread and methods abound in the field of drug development and pharmacological research. In this study, we choose rosiglitazone compound as the object to predict the targets and to discuss the mechanism based on three kinds of prediction methods of network pharmacology. Comparison of the prediction result has identified that the three kinds of prediction methods had their own characteristics: targets and pathways predicted were not in accordance with each other. However, the calcium signaling pathway could be predicted in the three kinds of methods, which associated with diabetes and cognitive impairment caused by diabetes by bioinformatics analysis. The above conclusion indicates that the calcium signaling pathway is important in signal pathway regulation of rosiglitazone compound, which provides a clue to further explain the mechanism of the compound and also provides a reference for the selection and application of methods of network pharmacology in the actual research.
Calcium Signaling ; Cognitive Dysfunction ; Computational Biology ; Diabetes Mellitus ; Humans ; Pharmacology ; methods ; Thiazolidinediones ; pharmacology

Objective To investigate the expression of osteopontin (OPN)in serum,urine and renal tissue of patients with systemic lupus erythematosus (SLE)and its relevance with organ damage in and activity of this disease.Methods Enzyme-linked immunoabsorbent assay(ELISA)was applied to detect the concentration of OPN in the sera of 100 patients with SLE and 30 sex-and age-matched normal human controls as well as in the urine of 57 patients with SLE and 15 normal human controls.Renal tissue was obtained from 3 patients with lupus nephritis and subjected to immunohistochemistry for the observation of OPN.Results The level of OPN was significantly higher in the sera and urine of patients with SLE than in those of normal human controls (64.03 ±72.87 μg/L vs 29.88±1 1.28μg/L,454.87±231.63 μg/L vs 122.67±39.47μg/L,both P<0.05).Increased level of OPN in sera and urine was also observed in patients with active SLE com-pared with those with inactive SLE (80.92±87.49μg/L vs 36.43±23.48μg/L,584.36±207.15 μs/L vs 28 1.08±1 3 1.92μg/L,both P<0.05).A positive correlation was noted between the serum level of OPN and SLE disease activity index(SLEDAI)(r=0.462,P<0.01),and the level of urine OPN was positively correlated with both SLEDAI and the concentration of urine immunoglobulin G.urine mierualbumin.urine α1-microglo-bulin,urine β2-microglobulin(r=0.901,0.458,0.359,0.342,0.409,respectively,all P<0.05).OPN was found in renal tubule epithelia of the three patients with lupus nephritis.Conclusion OPN may be involved in the pathogenesis of SLE and associated with the renal damage in patients with lupus nephritis.