Objective To elucidate the molecular mechanism of sirtuin-3 (SIRT3) in regulating mitochondrial biogenesis in human renal tubular epithelial cells. Methods Cells were stimulated with different concentrations of H₂O₂ and divided into four groups: control (NC), 50 μmol/L H₂O₂, 110 μmol/L H₂O₂ and 150 μmol/L H₂O₂. SIRT3 protein expression was then measured. SIRT3 was knocked down with siRNA, and cells were further assigned to five groups: control (NC), negative-control siRNA (NCsi), SIRT3-siRNA (siSIRT3), NCsi+H₂O₂, and siSIRT3+H₂O₂. After 24 h, cellular adenosine triphosphate (ATP) and mitochondrial superoxide anion (O₂•⁻) levels were determined, together with mitochondrial expression of SIRT3, peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor 1 (NRF1), mitochondrial transcription factor A (TFAM), superoxide dismutase 2 (SOD2), acetylated-SOD2 and adenosine monophosphate activated protein kinase α1 (AMPKα1). Results The 110 and 150 μmol/L H₂O₂ decreased SIRT3 protein (both P<0.05). ATP and mitochondrial O₂•⁻ did not differ between NC and NCsi groups (both P>0.05). Compared to the NCsi group, the siSIRT3 group exhibited elevated O₂•⁻ level, decreased SIRT3 protein and increased expression levels of SOD2 and acetylated SOD2 protein (all P<0.05). Compared to the NCsi group, the NCsi+H₂O₂ group exhibited decreased cellular ATP levels, elevated mitochondrial O₂•⁻ levels, and reduced protein expression levels of SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 (all P<0.05). Compared with the siSIRT3 group, the siSIRT3+H₂O₂ group showed a decrease in cellular ATP levels, an increase in mitochondrial O₂•⁻ levels, a decrease in SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 protein expression levels and a decrease in acetylated SOD2 protein expression levels (all P<0.05). Compared with the NCsi+H₂O₂ group, the siSIRT3+H₂O₂ group showed a decrease in cellular ATP levels, an increase in mitochondrial O₂•⁻ levels, a decrease in SIRT3, AMPKα1, PGC-1α and NRF1, TFAM protein expression levels, and an increase in SOD2 and acetylated SOD2 protein expression levels (all P<0.05). Conclusions SIRT3 promotes mitochondrial biogenesis in tubular epithelial cells via the AMPK/PGC-1α/NRF1/TFAM axis, representing a key mechanism through which SIRT3 ameliorates oxidative stress-induced mitochondrial dysfunction.

Objective To investigate the effect on blood coagulation function and immune function in patients with gas-tric cancer by laparoscopic radical gastrectomy and open operation. Methods In our hospital from June 2011 to June 2014,selected hospitalized patients with gastric cancer 30 cases as the research object,15 cases were undergone tradi-tional open operation in patients with the establishment of the control group, the remaining 15 cases received laparo-scopic radical resection of gastric cancer,set up the observation group,the changes of the TT,APTT,INR,FIB,D-D of blood coagulation indexes of two groups were compared,and changes in two groups of patients with immune function indexes of CD3+,CD4+,CD8+,CD4+/CD8+,and the operation time,bleeding amount of two groups of operation,ambula-tion time, hospital stay, postoperative complications were compared between two groups. Results In the observation group and the control group after 24 h,the FIB,D-D was significantly higher than the preoperative,and the observa-tion group patients after operation significantly were higher than the control group (P<0.05). After operation 1 weeks,CD3+,CD4+, CD8+, CD4+/CD8+of observation group showed no significant changes (P>0.05). CD3+, CD4+, CD4+/CD8+of control group after 1 weeks significantly changed compared with the preoperative, and CD3+, CD4+, CD4+/CD8+ of observation group compared with the control group,the difference was statistically significant(P<0.05). But CD8+of the control group had no obvious change before and after surgery(P>0.05). The observation group patients' out of bed activity time, postoperative hospitalization time was significantly shorter than that in the control group,the amount of bleeding observation group was significantly less than that the control group(P<0.05). Conclusion Laparoscopic radical gastrectomy for gastric cancer curative has good effect, compared with open radical gastrectomy for gastric cancer with less bleed-ing, fewer complications, patients recovered quickly, the advantages of immune function in patients with small inter-ference, but the effects of laparoscopic operation on blood coagulation in patients with more significant, corresponding measures should be taken to prevent the formation of thrombus coagulation abnormalities caused by postoperative.

The cDNA of Insulin-like growth factor binding protein 3 was subcloned into a eukaryotic secretory expression vector pSectagA to construct pSectag-IGFBP3. Human renal cell carcinoma (RCC) 786-0 cells were transfected with pSectag-IGFBP3 using lipofectamine 2000. After 48 h, the secretory IGFBP-3 was tested and identified by western blotting. Meanwhile, Annexin V-EGFP stain was used to analyze the apoptosis of 786-0 cells induced by IGFBP-3. Secretory IGFBP-3 protein could express successfully in the 786-0 cells and the expressed IGFBP-3 directly displayed an apoptotic effect on the host cells. This work provides a basis for further study on the apoptosis-inducing mechanism of IGFBP-3 and the development of a new anti-tumor drug.
Apoptosis ; drug effects ; Base Sequence ; Carcinoma, Renal Cell ; metabolism ; pathology ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; genetics ; secretion ; Kidney Neoplasms ; metabolism ; pathology ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; physiology ; Transfection ; Tumor Cells, Cultured