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OBJECTIVE To explore the effects and mechanism of atorvastatin on the proliferation， autophagy and glucose metabolism of AGS cells in human gastric cancer. METHODS The effects of low， medium and high concentrations of atorvastatin （12.5， 25， 50 μmol/L） on the viability of AGS cells were investigated through preliminary experiments， and the concentration of action was screened. The formal experiment was divided into control group （no intervention）， atorvastatin group （25 μmol/L）， positive control group （50 mg/L 5-fluorouracil）， inhibitor group [25 μmol/L atorvastatin +10 μmol/L phosphatidylinositol-3-kinase （PI3K）/protein kinase B （Akt） signaling pathway inhibitor LY294002] and activator group （25 μmol/L atorvastatin +10 μmol/L PI3K/Akt signaling pathway activator SC79）， all of which were treated for 24 h. Glucose metabolism （glucose and lactic acid contents） and cell proliferation rate were detected， as well as the expression of autophagy-associated protein light chain 3 （LC3） Ⅰ， LC3Ⅱ and PI3K/Akt signaling pathway-associated proteins in cells. RESULTS Both medium and high concentrations of atorvastatin could significantly inhibit the viability of AGS cells （P＜0.05）， and 25 μmol/L atorvastatin was selected for the official experiment for follow-up experiments. Compared with the control group， the contents of glucose and lactic acid， cell proliferation rate， p-PI3K/PI3K and p-Akt/Akt ratios in the positive control group and atorvastatin group were significantly decreased （P＜ 0.05）， and the protein expression levels of LC3 Ⅰ and LC3 Ⅱ were significantly increased （P＜0.05）. Compared with the atorvastatin group， the inhibitor further promoted the changes in the above indexes （P＜0.05）， and the activator significantly reversed the changes in the above indexes （P＜0.05）. CONCLUSIONS Atorvastatin could inhibit glucose metabolism and proliferation of AGS cells in human gastric cancer and promote autophagy. The mechanism may be related to the inhibition of the PI3K/Akt signaling pathway.
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Objective To investigate the relationship between the systemic lupus erythematosus (SLE)and the SOCS-1 gene methylation status of the peripheral blood DNA,to provide the basis for diagnosis and treatment of systemic lupus erythema-tosus.Methods Blood samples of SLE patients (27 cases)and healthy group (19 cases)in January 2015 to April were col-lected and the DNA were extracted.Using polymerase chain reaction combining DNA agarose gel electrophoresis to detect the SOCS-1 gene methylation status.Results In patients with systemic lupus erythematosus SOCS-1 gene complete methyl-ation accounted for 44% (12/27),incomplete methylation accounted for 56% (15/27).In healthy group SOCS-1 gene com-plete methylation accounted for 74% (14/19)and incomplete methylation accounted for 26% (5/19).The rate of complete methylation of SOCS-1 gene of SLE patients was lower than that of healthy group (χ2=3.88,P=0.049).Conclusion SLE patients may have lower SOCS-1 gene methylation status in the peripheral blood DNA,which is worth for further study.
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Objective To analyze the epidemiological characteristics of Epstein Barr virus (EBV)or cytomegalovirus (CMV) infection in systemic lupus erythematosus (SLE)patients in order to provide reference for clinic.Methods The clinical data of 202 female cases in-patients diagnosed with SLE (SLE group)from January 2012 to May 2015 in Nanjing Drum Tower Hospital were analyzed retrospectively.Meanwhile,203 cases including female renal transplant donors,obstetrics and gyne-cology in-patients selected randomly were enrolled as control group.The infection rate between SLE and control groups was analyzed and compared based on the results of EBV-DNA and CMV-DNA in peripheral blood quantified by real time PCR. Results The positive rate of EBV-DNA in SLE group was 11.39% (23/202),with significantly statistical difference when comparing with the control group [3.45% (6/174)](χ2 = 8.28,P < 0.01).The positive rate of CMV-DNA [7.92% (16/202)]was significantly higher than that in control group [1.97% (4/203)](χ2 =7.64,P <0.05).In addition,the positive rate of EBV-DNA and CMV-DNA was also greatly higher in reproductive-age (20~45 years old)of SLE patients than those in control group,10.94%(14/128)vs 3.45%(4/116)(χ2 =4.99,P <0.05)and 7.75%(10/129)vs 2.22%(3/135)(χ2 =4.31,P <0.05),respectively.Conclusion SLE patients were more inclined to be accompanying infected by EBV or CMV, indicating the possible correlation between SLE and EBV or CMV infection;and physicians should pay more attention to the viral infection of SLE in clinical treatment.
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Objective To preliminarily investigate the effect of ribosomal protein S7 on apoptosis of HeLa cervical cancer cells.Methods The previously constructed recombinant plasmid pIRES2-EGFP-RPS7 was transfected into HeLa cells,the empty vector pIRES2-EGFP transfected cells as control.Enhanced green fluorescent protein(EGFP)expressing cells were quantified by flow cytometry,and RPS7 protein level was also determined by Western blotting.Cell apoptosis of both RPS7 over-expression cells and knockdown cells were evaluated by flow cytometry after staining using allophycocyanin labeled Annexin-V.Results Apoptotic cell level in the obtained RPS7 transient over-expression HeLa cells was significantly higher than that of vector con-trol cells [(1 0.00 ±0.60)% vs (5.73 ±0.61 )%],with a statistic difference (t =8.63,P =0.001 ). Moreover,the apoptotic level in RPS7 knockdown cells was lower than that in control cells [(3.08 ± 0.49)% vs (5.97 ±0.63)%],with a statistic difference (t =6.40,P =0.003).Conclusion Up-regula-tion of RPS7 may promote apoptosis,while down-regulation of RPS7 may inhibit apoptosis of HeLa cells,indi-cating that RPS7 may play roles in regulating cell apoptosis.
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Objective To compare the results of anti-cardiolipin (ACA)measuring by two commercial available kits.Methods ACA in total of 66 serum samples were both determined by kits from Euroimmun and YHLO simultaneously,then the re-sults were analyzed comparatively and correlatively.The Euroimmun kit was applied to determination the level of ACA-IgA/G/M,and the YHLO kit determined ACA-IgG/M and anti-β2-glycoprotein I antibody (β2GPI IgG).Results The positive rate by Euroimmun kit was 37.88% (25/66),while 31.82% (21/66)was positive (one positive among ACA-IgG,IgM andβ2GPI IgG)when determined by YHLO kit,and there was no significant difference between the two kits.The accordance rate of the two kits was 87.88% (58/66).The ACA-IgA/G/M value by Euroimmun kit and the summation of ACA-IgG, IgM andβ2GPI IgG by YHLO kit showed well linear correlation (r 2 =0.892,P <0.01).Conclusion Results from the two kits were consistent and correlated well,and they are suitable for the clinical application;these two kits have their own char-acteristics,which could be used by individual or combination accordingly.