Nitrites play multiple characteristic functions in invasion and metastasis of hepatic cancer cells, but the exact mechanism is not yet known. Cancer cells can maintain the malignant characteristics via clearance of excess mitochondria by mitophagy. The purpose of this article was to determine the roles of nitrite, reactive oxygen species (ROS) and hypoxia inducing factor 1 alpha (HIF-1 α) in mitophagy of hepatic cancer cells. After exposure of human hepatocellular carcinoma SMMC-7721 cells to a serial concentrations of sodium nitrite for 24 h under normal oxygen, the maximal cell vitality was increased by 16 mg x (-1) sodium nitrite. In addition, the potentials of migration and invasion for SMMC-7721 cells were increased significantly at the same time. Furthermore, sodium nitrite exposure displayed an increase of stress fibers, lamellipodum and perinuclear mitochondrial distribution by cell staining with Actin-Tracker Green and Mito-Tracker Red, which was reversed by N-acetylcysteine (NAC, a reactive oxygen scavenger). DCFH-DA staining with fluorescent microscopy showed that the intracellular level of ROS concentration was increased by the sodium nitrite treatment. LC3 immunostaining and Western blot results showed that sodium nitrite enhanced cell autophagy flux. Under the transmission electron microscopy (TEM), more autolysosomes formed after sodium nitrite treatment and NAC could prevent autophagosome degradation. RT-PCR results indicated that the expression levels of COX I and COXIV mRNA were decreased significantly after sodium nitrite treatment. Meanwhile, laser scanning confocal microscopy showed that sodium nitrite significantly reduced mitochondrial mass detected by Mito-Tracker Green staining. The expression levels of HIF-1α, Beclin-1 and Bnip3 (mitophagy marker molecular) increased remarkably after sodium nitrite treatment, which were reversed by NAC. Our results demonstrated that sodium nitrite (16 mg x L(-1)) increased the potentials of invasion and migration of hepatic cancer SMMC-7721 cells through induction of ROS and HIF-1α mediated mitophagy.

Objective:To explore the clinical efficacy of single-center infant kidney donor adult dual kidney transplantation to explore the difference of different operation methods and the operation options of different donor kidney conditions so as to improve the success rate of children kidney donor adult dual kidney transplantation and reduce complications.Methods:A total of 42 cases of infant and adult dual kidney transplantations at Department of Kidney Transplantation in The Second Xiangya Hospital of Central South University from December 2012 to May 2019 were divided into two groups according to whether or not donor kidney fulfilled the criteria of three " 5" . According to different surgical approaches, they were divided into three groups of A (classical En-Bloc operation), B (separated dual kidney transplantation) and C (modified operation). The clinical data and prognoses were analyzed.Results:The median follow-up period was 55(11-92) months. The estimated glomerular filtration rate was 123.4(92.2-156.6) ml/min for operation A, 97.2(81.3-116.6) ml/min for operation B and 133.9(133.9-133.9) ml/min for operation C. In donor group not fulfilling the " 5" principle, no thrombotic event occurred for operation A/C and 3 cases of transplantation for operation B. There were single renal embolism ( n=2) and dual renal embolism ( n=1)(3/5, 60%)( P<0.05). Urinary protein was positive in the last follow-up: operation A (1/2, 50%) and operation B (3/5, 60%) ( P<0.05). The estimated glomerular filtration rate at the last follow-up was 82.4(80.9-83.9) ml/min for operation A, 71.8(46.1-114.2) ml/min for operation B and 122(83.3-142.4) ml/min for operation C. The 1-year graft survival rate was 100% and 89.5% in three " 5" donor group and 3-year graft survival rate was 100% and 84.2% respectively. Conclusions:Satisfactory outcomes might be obtained during dual kidney transplantation for infants and adults. The incidence of thrombosis, urine leakage and urinary protein is lower in improved kidney transplantation group than that in previously operated group. The problem of graft hyperperfusion injury is well solved. And the long-term follow-up outcome is excellent.

Adrenal Gland Diseases ; diagnosis ; Female ; Humans ; Middle Aged ; Retroperitoneal Space