To study the mechanisms whereby cisplatin suppresses survival of human esophageal squamous cell carcinoma cells.The cytotoxicity of cisplatin in cisplatin-resistant cell line EC109 /CDDP and its parental cell line EC109 was measured by cell viability assay.Western blotting was used to investigate the protein expression of to-tal p53 and phosphorylated p53 at Ser15.Colony formation assay was employed to evaluate the ability of cells to recover from treatments and form colonies.The results indicated that EC109 /CDDP cells were more resistant to cisplatin-induced cytotoxicity than EC109 cells,with the IC50 values of (20.4 ±4.4)μmol /L and (5.7 ±0.1 )μmol /L,respectively.Although cisplatin did not alter the total protein level of p53,it obviously increased the phosphorylation of p53 at Ser15.Cisplatin inhibited survival of both EC109 /CDDP and EC109.Notably,inhibition of p53 by Pifithrin-αsignificantly promoted recovery of cisplatin-treated EC109 and EC109 /CDDP cells to differ-ent degrees.In this respect,p53 protein was found to be activated in response to cisplatin treatment in both EC109 /CDDP and EC109,which may contribute to the cytotoxic effect of cisplatin.

Objective To investigate the efficacy of sophocarpine in rats with alcoholic liver disease and its effects on the expression of tumor necrosis factor (TNF)-α,interleukin (IL)-6 and transforming growth factor (TGF)-β1.Methods A total of 48 male Sprague-Dawley adult rats were evenly divided into healthy control group,model group,prevention group and treatment group.The rats in the healthy control group were gavaged with 0.9％NaCl every day for 12 weeks.The rats in the model group,prevention group and treatment group were gavaged with alcohol for 12 weeks to establish the model.The prevention group was injected with 20 mg · kg1 · d1 sophocarpine for 12 weeks.Since the fifth week,the treatment group was continuously injected with 20 mg · kg1 · d-1 sophocarpine for eight weeks.The histological changes were evaluated.The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase ( AST ), alkaline phosphatase (AKP),triglyceride (TG) and total cholesterol (TC) were examined.And the expression of TNF-α,IL-6 and TGF-β1 in liver tissue at mRNA and protein level were detected with immunohistochemistry and real-time polymerase chain reaction (PCR) assay.Comparison among groups was perform with single factor analysis of variance,pairwise comparisons with least significant difference method (LSD method),ranked data with Kruskal-Wallis H-test and multiple pairwise comparison with Nemenyi test.Results Compared with model group,hepatic steatosis and inflammatory cell infiltration were significantly improved in the treatment group and prevention group.The levels of ALT (41.40 U/L± 10.53 U/L and 40.75 U/L±6.94 U/L vs 58.37 U/I±5.35 U/L),AST(121.60 U/L±16.24 U/L and 109.50 U/L±9.23 U/L vs 156.63 U/L±32.47 U/L),AKP(114.88 U/L±40.37 U/L and 112.60 U/L±44.34 U/L vs 161.75 U/L±28.95 U/L),TG (4.19 mmol/L±0.99 mmol/L and 2.69 mmol/L± 1.35 mmol/L vs 4.50 mmol/L±0.99 mmol/L) and TC (1.48 mmol/L±0.28 mmol/L and 1.43 mmol/L±0.19 mmol/L vs 1.67 mmol/L±0.20 mmol/L) significantly decreased and the difference was statistically significant ( all P＜0.05).The expression of TNF-α,IL-6 and TGF-β1 at mRNA and protein level in liver tissue of model group were significantly higher than those of healthy control group,prevention group and treatment group.After treated with sophocarpine,the expression of TNF-α(mRNA:1.36 ± 0.08,1.16 ± 0.05 ; protein:3.38 ％ ± 0.82 ％,1.74 ％ ± 0.65 ％ ),IL-6 (mRNA:1.51 ± 0.05,1.39 ± 0.02; protein:5.89％ ± 0.96％,4.26％ ± 0.53％) and TGF-β1 (mRNA:1.39±0.04,1.37±0.02; protein:4.27％ ±0.97％,2.11％ ±0.83％) of treatment group and prevention group at mRNA and protein level significantly lower than those of model group (mRNA:1.81±0.16,1.95 ±0.13,1.84±0.22; protein:5.82％ ± 1.21％,7.63％ ±1.03％,5.33％± 1.12％) and the difference was statistically significant (all P＜0.01).Conclusion Sophocarpine significantly alleviates alcohol induced liver injury in rats,improves liver steatosis and inflammatory reaction degree,which may be related with the downregulation of TNF-α,TGF-β1 and IL-6 expression in liver tissue of ALD rats.

OBJECTIVETo explore the antagonizing effect of celecoxib against the cytotoxicity of carboplatin in human esophageal cancer cells.

METHODSThe cell viability of cisplatin-resistant cell line EC109/CDDP and its parental cell line EC109 exposed to carboplatin alone or carboplatin plus celecoxib was determined by MTT assay. The expression of CTR1, caspase-3 activation and PARP cleavage in the exposed cells were examined by Western blotting. Caspase-3 activity and cell apoptosis after the exposure were detected with Caspase-3/7 assay and flow cytometry, respectively. The effect of celecoxib on carboplatin accumulation in the cells was measured using inductively coupled plasma mass spectrometry (ICP-MS).

RESULTSCelecoxib treatment significantly increased the IC50 of carboplatin, suppressed carboplatin-induced caspase-3 and PARP cleavage and caspase-3 activity in EC109 and EC109/CDDP cells. Celecoxib also inhibited carboplatin-induced apoptosis and suppressed intracellular carboplatin accumulation in both cell lines. A combined exposure to celecoxib and carboplatin did not cause significant changes in the protein expression of CTR1.

CONCLUSIONCelecoxib antagonizes the cytotoxic effect of carboplatin and inhibits carboplatin-induced apoptosis in human esophageal cancer cells by reducing intracellular carboplatin accumulation.

Apoptosis ; Blotting, Western ; Carboplatin ; antagonists & inhibitors ; Caspase 3 ; metabolism ; Celecoxib ; Cell Line, Tumor ; drug effects ; Cell Survival ; Drug Interactions ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Pyrazoles ; pharmacology ; Sulfonamides ; pharmacology

Carotid Artery, Internal, Dissection ; therapy ; Humans ; Male ; Middle Aged ; Treatment Outcome

Humans ; Male ; Middle Aged ; Polypropylenes ; Reconstructive Surgical Procedures ; Respiratory Tract Fistula ; surgery ; Surgical Mesh

Objective To explore the roles of serum TLR-4, TNF-αand IL-6 in neonates with preterm birth. Methods A total of 120 neonates from neonatology department in the Xingtai People's Hospital were selected and divided into full-term group (n=40), premature rupture of fetal membranes (n=40) and idiopathic preterm group (n=40) based on the gestational age. The peripheral venous blood was collected within 30 minutes when the infants were born, and the supernatant was reserved after centrifuged. The levels of serum TLR-4, TNF-αand IL-6 were detected by enzyme-linked immunosorbent assay. Results The levels of TLR-4, TNF-αand IL-6 in idiopathic preterm and premature rupture of fetal membranes were signiifcantly higher than that in full-term group and showed positive correlation. Conclusion Cytokines TLR-4, TNF-αand IL-6 maybe closely related to the preterm birth.

Objective To explore the antagonizing effect of celecoxib against the cytotoxicity of carboplatin in human esophageal cancer cells. Methods The cell viability of cisplatin-resistant cell line EC109/CDDP and its parental cell line EC109 exposed to carboplatin alone or carboplatin plus celecoxib was determined by MTT assay. The expression of CTR1, caspase-3 activation and PARP cleavage in the exposed cells were examined by Western blotting. Caspase-3 activity and cell apoptosis after the exposure were detected with Caspase-3/7 assay and flow cytometry, respectively. The effect of celecoxib on carboplatin accumulation in the cells was measured using inductively coupled plasma mass spectrometry (ICP-MS). Results Celecoxib treatment significantly increased the IC50 of carboplatin, suppressed carboplatin-induced caspase-3 and PARP cleavage and caspase-3 activity in EC109 and EC109/CDDP cells. Celecoxib also inhibited carboplatin-induced apoptosis and suppressed intracellular carboplatin accumulation in both cell lines. A combined exposure to celecoxib and carboplatin did not cause significant changes in the protein expression of CTR1. Conclusion Celecoxib antagonizes the cytotoxic effect of carboplatin and inhibits carboplatin-induced apoptosis in human esophageal cancer cells by reducing intracellular carboplatin accumulation.

Objective To study the effect of oncogenic Ras overexpression on autophagic activity in human fibroblast cells in vitro. Methods BJ cells were transfected with H-RasV12 or control vector and treated with chloroquine, small interfering RNA (siRNA) for ATG7, or rapamycin. The cellular responses were analyzed by monitoring the parameters and biomarkers for cell growth, senescence and cell death. Results In BJ cells overexpressing H-RasV12, chloroquine treatment resulted in more prominent cell senescence and a significantly increased cell death rate. Suppression of ATG7 mediated by siRNA also promoted cell senescence. Rapamycin treatment also caused an increased cell death rate but attenuated senescence in surviving cells. In control BJ cells, the cellular response to chloroquine included senescence and cell death, which occurred slowly. Rapamycin treatment and siRNA suppression of ATG7 had no obvious effect on control BJ cells. Conclussion Stable cellular overexpression of oncogenic Ras causes tightly controlled suppression of the autophagic activity of human fibroblast cells, and such changes produce significant effect on cell senescence and survival.

Objective To explore the antagonizing effect of celecoxib against the cytotoxicity of carboplatin in human esophageal cancer cells. Methods The cell viability of cisplatin-resistant cell line EC109/CDDP and its parental cell line EC109 exposed to carboplatin alone or carboplatin plus celecoxib was determined by MTT assay. The expression of CTR1, caspase-3 activation and PARP cleavage in the exposed cells were examined by Western blotting. Caspase-3 activity and cell apoptosis after the exposure were detected with Caspase-3/7 assay and flow cytometry, respectively. The effect of celecoxib on carboplatin accumulation in the cells was measured using inductively coupled plasma mass spectrometry (ICP-MS). Results Celecoxib treatment significantly increased the IC50 of carboplatin, suppressed carboplatin-induced caspase-3 and PARP cleavage and caspase-3 activity in EC109 and EC109/CDDP cells. Celecoxib also inhibited carboplatin-induced apoptosis and suppressed intracellular carboplatin accumulation in both cell lines. A combined exposure to celecoxib and carboplatin did not cause significant changes in the protein expression of CTR1. Conclusion Celecoxib antagonizes the cytotoxic effect of carboplatin and inhibits carboplatin-induced apoptosis in human esophageal cancer cells by reducing intracellular carboplatin accumulation.

Objective To study the effect of oncogenic Ras overexpression on autophagic activity in human fibroblast cells in vitro. Methods BJ cells were transfected with H-RasV12 or control vector and treated with chloroquine, small interfering RNA (siRNA) for ATG7, or rapamycin. The cellular responses were analyzed by monitoring the parameters and biomarkers for cell growth, senescence and cell death. Results In BJ cells overexpressing H-RasV12, chloroquine treatment resulted in more prominent cell senescence and a significantly increased cell death rate. Suppression of ATG7 mediated by siRNA also promoted cell senescence. Rapamycin treatment also caused an increased cell death rate but attenuated senescence in surviving cells. In control BJ cells, the cellular response to chloroquine included senescence and cell death, which occurred slowly. Rapamycin treatment and siRNA suppression of ATG7 had no obvious effect on control BJ cells. Conclussion Stable cellular overexpression of oncogenic Ras causes tightly controlled suppression of the autophagic activity of human fibroblast cells, and such changes produce significant effect on cell senescence and survival.