To investigate the potential ability of the nitrite to induce neuronal differentiation of PC12 cells, cultured PC12 cells planted on matrigel in the presence or absence of sodium nitrite were employed as model, nerve growth factor (NGF) served as a positive control. After 48 h, sodium nitrite enhanced cell viability and vascular endothelial growth factor (VEGF) secretion. Same as the effect of NGF, sodium nitrite (1.4 mmol x L(-1)) treated cultures contained a greater proportion of cells bearing neurites and neurites were much longer than those found in negative control cultures (P < 0.05). Compared with the negative control, sodium nitrite (1.4 mmol x L(-1)) also upregulated the expression of VEGF mRNA (P < 0.05) and hypoxia inducible factor 1 alpha (HIF-1 alpha) or VEGF protein expression (P < 0.05) in cultures of PC12 cells. On the other hand, these effects of the sodium nitrite were likely mediated by HIF-1alpha, since their effects were antagonized by addition of HIF-1alpha inhibitor YC-1. Taken together, these results suggest that low doses of sodium nitrite could induce neurite outgrowth in PC12 cells by activating the HIF-1alpha-VEGF pathway.
Animals ; Cell Differentiation ; drug effects ; Cell Survival ; drug effects ; Food Preservatives ; pharmacology ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Neurites ; drug effects ; PC12 Cells ; RNA, Messenger ; metabolism ; Rats ; Sodium Nitrite ; pharmacology ; Up-Regulation ; Vascular Endothelial Growth Factor A ; genetics ; secretion

This study is to find out the induction by sodium nitrite of epithelial-mesenchymal transition (EMT) in human hepatocellular carcinoma cells, SMMC-7721. After treatment of SMMC-7721 with 0.25 - 25 mmol.L-1 sodium nitrite for 48 h, the assays used include enzyme-linked immunosorbent assay (ELISA) for evaluation of TGF-beta1, IL-6 and IL-8 level in the conditioned medium, phase-contrast microscopy for morphology observation, and scratch wound healing as well as transwell migration assays for measurement of migration and metastatic potential. Additionally, the hallmarks of EMT, p-AKT and its downstream signaling molecules were examined by Western blotting. The results showed that TGF-beta1 secreted by SMMC-7721 elevated significantly in a dose-dependent fashion, whereas the increased IL-8 and IL-6 did not show dose-dependent response. The EMT was induced by exposure of SMMC-7721 with 0.25 mmol.L-1 of sodium nitrite, which was characterized by increased level of Vimentin, decreased E-cadherin and elevated activity of migration and metastatic potential. The results suggest that sodium nitrite could induce SMMC-7721 EMT by increased secretion of TGF-beta1 and IL-8.
Cadherins ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; Dose-Response Relationship, Drug ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Liver Neoplasms ; metabolism ; pathology ; NF-kappa B ; metabolism ; Neoplasm Invasiveness ; Proto-Oncogene Proteins c-akt ; metabolism ; Sodium Nitrite ; administration & dosage ; pharmacology ; Transforming Growth Factor beta1 ; secretion ; Twist-Related Protein 1 ; metabolism ; Vimentin ; metabolism

This study is to observe the effect of N-(3-phenylallylidene)-6-fluoro-1, 8-(2, 1-propoxy)-7-(4-methylpiperazin-1-yl)-quinolin-4(1H)-one-3-carbonyl hyarazine (FQ16) on apoptosis of hepatocarcinoma SMMC-7721 cells in vitro. With different concentrations of FQ16 at different times used to treat SMMC-7721 cells in vitro, the proliferation of the cells and the inhibition effect of FQ16 on the cell proliferation were examined by MTT assay. Cell apoptosis was determined by Hoechst 33258/PI fluorescence staining, TUNEL and agarose gel electrophoresis method. The effect of FQ16 on topoisomerase II activity was measured by agarose gel electrophoresis using Plasmid pBR322 DNA as the substrate. Mitochondrial membrane potential (MMP, delta psi m) was measured by high content screening image system. The reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression changes of Bcl-2 mRNA and Bax mRNA. The caspase-9, caspase-8, caspase-3, p53, Bcl-2 and Bax protein expressions were detected by Western blotting analysis. The results showed that the cell proliferation was inhibited by FQ16 at 0.625 - 10 micromol L(-1) in a time-dose dependent manner. Treatment of SMMC-7721 cells with different concentrations of FQ16 for 24 h increased the percentage of the apoptosis cells obviously (P<0.05), the typical ladder DNA in apoptotic cells and a concomitant dissipation of the mitochondrial membrane potential. Compared with control group, FQ16 influenced obviously DNA topoisomerase II activity, stimulated DNA cleavage and inhibited DNA reunion mediated by topoisomerase II. In addition, FQ16 (3 - 7.39 micromol L(-1)) increased mRNA expression of Bax and protein expression of p53, Bax, caspase-9, caspase-3, separately, and induced cytosolic accumulation of activities caspase-9 and caspase-3, whereas the mRNA and protein expression of Bcl-2 decreased with no change of caspase-8. Therefore it can be concluded that the effects of inhibited topoisomerase II and mitochondrial-dependent pathways were involved in FQ16 induction of apoptosis of SMMC-7721 cells.
Antineoplastic Agents ; administration & dosage ; chemical synthesis ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Topoisomerases, Type II ; metabolism ; Dose-Response Relationship, Drug ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Membrane Potential, Mitochondrial ; drug effects ; Molecular Structure ; Piperazines ; administration & dosage ; chemical synthesis ; chemistry ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; bcl-2-Associated X Protein ; metabolism

This study is to investigate the cytoprotective role of NaNO2 preconditioning against ethanol induced damage in human hepatoma SMMC-7721 cells. The cells were preconditioned with NaNO2 (0.25 mmol x L(-1)) for 24 hours or 4 weeks, and then exposed to ethanol (200 mmol x L(-1)) for additional 12 h and untreated cells served as control. Both temporal and chronic NaNO2 preconditioning could prevent ethanol elicited cytotoxicity as evidenced by thiazolyl blue (MTT). NaNO2 preconditioning also could inhibit ethanol-induced apoptosis, which was confirmed by FITC-Annexin V/PI flow cytometer and Hoechst 33258 and PI staining. Further, simultaneous NaNO2 preconditioning treatment along with ethanol showed protection against ethanol mediated cellular damage as indicated by significantly decreased levels of malondialdehyde (MDA) and elevated activities of superoxide dismutase (SOD) and catalase (CAT). Western blotting analysis revealed that in ethanol treated cells preconditioned with NaNO2, the HIF-1alpha and Bcl-2 increased obviously, while the expression of pro-apoptotic proteins, including Bax, Caspase-9, Caspase-3 decreased. The results showed that low doses of NaNO2 preconditioning resistant to ethanol-induced human hepatoma SMMC-7721 cells apoptosis, which mechanism may be related to increased expression of HIF-1alpha in the cells.
Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Catalase ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Ethanol ; toxicity ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Malondialdehyde ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Sodium Nitrite ; administration & dosage ; pharmacology ; Superoxide Dismutase ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo study the therapeutic effects of combined anterior-posterior (small incision or micro-incision) approach for complex tibial plateau and posterior condylar fractures.

METHODSFrom 2000 to 2008, 79 patients (81 limbs) with complex tibial plateau and posterior condylar fractures were reviewed. There were 45 males and 34 females, ranging in age from 19 to 66 years, with an average of 40.6 years. Thirty-nine limbs were treated using small incision through combined anterior-posterior approach, in which 13 limbs were Schatzker type IV, 15 limbs were type V ,and 11 limbs were type VI. Other 42 limbs were treated using micro-incision through combined anterior-posterior approach, in which 18 limbs were Schatzker type IV, 16 limbs were type V, and 8 limbs were type VI. The Rasmussen scores for knee joint and radio scores were used to evaluate therapeutic effects after the treatment. The complications such as cutaneous necrosis and incision infection were observed.

RESULTSAll the patients were followed up. According to Rasmussen criterion, in small incision group, 16 limbs got an excellent result, 13 good, 7 fair and 3 bad; in micro-incision group,above data were 19, 11, 8 and 4 respectively. Comparison between the two groups, P = 0.924. Comparison of complications such as cutaneous necrosis and incision infection: in small incision group,10 limbs had the complications, and in micro-incision group were 4 limbs; the occurrence rate of small incision group were higher than that of micro-incision group (P = 0.047).

CONCLUSIONThere are no significant differences between the two groups in the knee joint function rehabilitation; however, there is smaller rate for cutaneous necrosis and incision infection in micro-incision group.

Adult ; Aged ; Ankle Injuries ; surgery ; Case-Control Studies ; Female ; Fracture Fixation ; methods ; Humans ; Knee Joint ; physiopathology ; Male ; Middle Aged ; Postoperative Complications ; epidemiology ; Retrospective Studies ; Tibial Fractures ; surgery

This study is to report the determination of the effect of sodium nitrite induced oxygen species (ROS) on the epithelial-mesenchymal transition in hepatoma cells in mice bearing H22 and investigation of its role in hypoxia-inducible factor 1alpha (HIF-1alpha) in this process. Mice hepatocarcinoma cell line H22 was inoculated subcutaneously into right axillary of sixty male Kunming mice and then randomly divided into four groups: control group; low-dose sodium nitrite group (10 mg x kg(-1)), medium-dose sodium nitrite group (20 mg x kg(-1)) and high-dose sodium nitrite group (30 mg x kg(-1)). Sodium nitrite group was given (ig) sodium nitrite with 10-30 mg x kg(-1) x d(-1) for 21 days. Compared with control group, there was no obvious difference between the two groups in the volume or weight of xenografts, but in sodium nitrite treatment group, the activity of SOD and CAT decreased and contents of MDA or nitrite increased in tumor tissue of mice bearing H22; epithelial-mesenchymal transition (EMT) of hepatoma cells was induced, the EMT-phenotype tumors displayed a greater degree of local aggressiveness, with dissection through adjacent fascia and skeletal muscle. The increased expression of HIF-la and vimentin and declination of E-cadherin were confirmed by immunohistochemistry and Western blotting. These data indicate sodium nitrite treatment could improve the epithelial-mesenchymal transition of xenografts in mice bearing H22, which might relate to the fact that ROS mediated signal pathway increased the expression of HIF-1alpha.
Animals ; Cadherins ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Catalase ; metabolism ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Male ; Malondialdehyde ; metabolism ; Mice ; Neoplasm Transplantation ; Random Allocation ; Reactive Oxygen Species ; metabolism ; Signal Transduction ; drug effects ; Sodium Nitrite ; administration & dosage ; pharmacology ; Superoxide Dismutase ; metabolism ; Tumor Burden ; drug effects ; Vimentin ; metabolism

Sporadic malignant pheochromocytoma, a rare disease with poor prognosis, is always difficult to treat due in part to lack of effective agents. We presented three patients with advanced malignant pheochromocytoma treated by sunitinib, which indicates that sunitinib is an effective agent for this malignancy.
Adrenal Gland Neoplasms ; drug therapy ; Adult ; Female ; Humans ; Indoles ; therapeutic use ; Male ; Middle Aged ; Pheochromocytoma ; drug therapy ; Protein Kinase Inhibitors ; therapeutic use ; Pyrroles ; therapeutic use

Objective To evaluate the safety,feasibility of the application of "four-quadrant" tumor target-artery positioning in "zero-ischemia" retroperitoneal laparoscopic partial nephrectomy.Methods A retrospective study was conducted on 25 patients who underwent retroperitoneal laparoscopic partial nephrectomy from November 2016 to March 2017 by single surgeon.There were 15 males and 10 females with a mean age of (52.6 ± 11.9) years old.The mean diameter of the tumor was (3.8 ± 0.5) cm.The mean R.E.N.A.L.score was (9.0±1.3).The mean GFR of the operation side was (49.2±11.4) ml/(min · 1.73 m2) before the operation.All 25 patients underwent 3 Dimentional CT for the reconstruction of their renal models.The tumor target-artery was located in "four-quadrant" system,which based on the boundary between tumor and renal parenchyma.Perioperative outcomes including the estimated blood loss (EBL),operation time,complications and oncological outcomes were carefully collected and analyzed.Results Of all the 25 patients,21 patients underwent "zero-ischemia" retroperitoneal laparoscopic partial nephrectomy,the mean operation time was (106.1 ± 18.8) min,the mean EBL was (162.2 ±68.0) ml with no transfusion case.4 patients converted to conventional retroperitoneal laparoscopic partial nephrectomy,the operation time was 110-140 min,the EBL was 350-500 ml and the warm-ischemia time was 12-20 min respectively.The pathological outcome shows renal cell carcinoma in 24 patients and papillary renal cell carcinoma in 1 patient with all negative margins.The mean GFR of the operation side was (45.1 ± 10.2) ml/(min · 1.73 m2) after three months of the surgery and shows no statistical difference (P =0.268).Conclusion The application of the "four-quadrant" tumor target-artery positioning in " zeroischemia" retroperitoneal laparoscopic partial nephrectomy is a safety and feasible method in terms of accurate location of the target-artery,better postoperative renal function,low EBL and reasonable oncological outcomes.

Objective:To investigate the clinical efficacy of membrane induction technique in the treatment of postoperative infection of tibial plateau fractures in adults.Methods:A retrospective case series study was conducted to analyze the clinical data of 21 adult patients with postoperative infection of tibial plateau fractures treated with membrane induction technique from April 2013 to May 2017 in Southwest Hospital of Army Medical University. There were 19 males and two females, aged 19-60 years [(44.1±5.8)years]. There was one patient with type IV fractures, 14 with type V, and 6 with type VI according to the initial fracture typing by Schatzker's classification. There were three patients with infection period of within 3 weeks, 12 of 3-10 weeks, and 6 of over 10 weeks. All patients underwent two-stage operation using membrane induction technique to place cement in the bone defect area. After removal of internal fixation and thorough debridement, antibiotic cement and internal fixation plate were placed at stage I. Bone graft and reconstruction was performed at stage II. The infection indicators were recorded. Infection indices were monitored, including white blood cell count (WBC), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). Clearance of infection, bony union and complications were evaluated. Range of motion (ROM) and scoring of Hospital for Special Surgery (HSS) were used to evaluate the function of knee joint.Results:All patients were followed up for 12-62 months with an average of 23.5 months. Compared with 3 months after stage II, the indicators of infection at stage I showed that WBC was decreased from (10.6±2.3)×10 9/L to (6.7±3.5)×10 9/L, ESR decreased from (26.0±5.3)mm/h to (12.1±4.3)mm/h, and CRP decreased from (10.0±1.5)mg/L to (5.8±1.0)mg/L ( P<0.05). Infection was cleared in 17 patients after stage I operation, and the other 4 patients had infection recurrence, which were given stage I debridement again to control the infection. Two patients were treated with local flap transfer to cover the wound because of skin soft tissue defect after debridement. Another two patients underwent knee arthrodesis, and none was amputated. X-ray film indicated bony union in 21 patients at 46 months (mean, 4.5 months) after operation, and clinical bone healing was acquired in all 21 patients. One patient showed donor site infection. No nonunion, recurrence of infection after stage II, deep vein thrombosis or pulmonary embolism occured after the second stage. At the latest follow-up, ROM in patients with infection periods within 3 weeks and 3-10 weeks was singnificantly improved from [(95.2±10.4)° and (85.7±11.5)°] to [(120.2±10.5)° and (98.6±12.2)°] ( P<0.01), but not in patients with infection periods of over 10 weeks ( P>0.05). The HSS score in all patients was significantly improved after operation [(65.6±8.2)points vs. (82.0±6.6)points]( P<0.01). Conclusion:For adult patients with tibial plateau fracture, membrane induction technique can effectively control the postoperative infection, achieve clinical bone healing and improve the knee function.

OBJECTIVETo translate the English version of The Premature Ejaculation Diagnostic Tool (PEDT) into Chinese, evaluate its reliability and validity, and analyze its feasibility in the diagnosis of premature ejaculation (PE).

METHODSFollowing the forward-backward translation procedure, we developed the Chinese version of PEDT, which was then revised by andrologists and bilingual linguists. We enrolled subjects with or without PE from 15 urological or andrological clinics in China and obtained the information about their demographic characteristics, PEDT scores, and intra-vaginal ejaculation latency time (IELT). We evaluated the internal consistency of PEDT using Cronbach alpha, was examined its reliability and stability by test-retest analysis, analyzed its correlation with IELT by Spearman correlation analysis, and tested its sensitivity and specificity by receiver operating characteristic ( ROC) analysis.

RESULTSTotally, 570 PE patients (aged [30.66 ± 7.11] years) and 226 non-PE men (aged [33.01 ± 5.41] years) were recruited, with the mean IELT of (1.34 ± 0.54) min in the former and (11.09 ± 7.5) min in the latter group. The Cronbach's alpha of the Chinese version of PEDT was 0.79, and the test-retest correlation coefficient was 0.75 (P < 0.01). The PEDT score was negatively correlated with IELT (Spearman's p = -0.52, P < 0.01). When the cutoff value of PE diagnosis was defined as 7.5, the sensitivity and specificity of PEDT were 0.80 and 0.78, and when as 8.5, they were 0.72 and 0.89, respectively.

CONCLUSIONThe Chinese version of PEDT was demonstrated to have good internal consistency, reliability, and validity, as well as a high predictability for PE. It can be used as a reliable and convenient tool to screen PE among Chinese men.

Adult ; Aged ; Asian Continental Ancestry Group ; China ; Ejaculation ; Feasibility Studies ; Humans ; Language ; Male ; Middle Aged ; Premature Ejaculation ; diagnosis ; ROC Curve ; Reaction Time ; Reproducibility of Results ; Sensitivity and Specificity ; Translations