Prognostic significance of USP22 as an oncogene in papillary thyroid carcinoma.

Ubiquitin-specific protease 22 (USP22), a novel deubiquitinating enzyme, has been associated with metastasis, therapy resistance, and cell cycle progression. The purpose of this study was to investigate the expression level of USP22 in papillary thyroid carcinoma (PTC) samples and to evaluate its clinical significance in PTC patients. USP22 expression was examined in 30 fresh PTC tissues and paired adjacent noncancerous tissues by real-time quantitative RT-PCR. Immunohistochemistry for USP22 was performed on additional 156 PTC tissues. The clinical significance of USP22 expression was analyzed. We found that the expression levels of USP22 mRNA and protein in PTC tissues were both significantly higher than those in noncancerous tissues. Clinicopathological analysis showed that USP22 expression was significantly correlated with tumor size (p=0.036), extracapsular invasion (p=0.012), multifocality (p=0.014), lymph node metastasis (p=0.022), distant metastasis (p=0.005), and TNM stage (p=0.002). The Kaplan-Meier survival curves revealed that USP22 expression was associated with poor prognosis in PTC patients. USP22 expression was an independent prognostic marker of overall patient survival in a multivariate analysis. Our findings suggest that USP22 is an independent predictor of poor prognosis of PTC patients.

Postprocedure administration of insulin in canine autologous vein grafting: a potential strategy to attenuate intimal hyperplasia.

Intimal hyperplasia (IH) exerts a critical role in vein graft failure after arterial bypassing. Insulin has been demonstrated to remarkably decrease IH in the rat carotid injury model. We hypothesized that postoperative insulin medication prevents the autologous vein graft from IH. Dogs were subjected to jugular-carotid interposition bypass grafting and intravenously infused with vehicle, glucose-insulin-potassium, glucose-potassium, or glucose-insulin-potassium plus Wortmannin 5 minutes before and 4 hours after reperfusion. Then vein grafts were harvested for caspase-3 activation, cell apoptosis, phosphorylated Akt, and endothelial nitric oxide synthase level assays. Other dogs undergoing the same operation were administered with subcutaneous injection of 4 U insulin or 0.5 mL saline two times per day for 1 month postoperatively. Vein grafts were sampled to assess cell proliferation, intimal/medial thickness, and expression of endothelial nitric oxide synthase and [alpha]-smooth muscle actin. Glucose-potassium aggravated apoptosis and caspase-3 activation and decreased Akt and endothelial nitric oxide synthase phosphorylation; however, glucose-insulin-potassium significantly inhibited cell apoptosis and caspase-3 activation and increased phosphorylated Akt and pendothelial nitric oxide synthase levels in canine vein grafts. Wortmannin largely abolished the glucose-insulin-potassium-elicited effects. Moreover, postoperative insulin use greatly inhibited cell proliferation, reduced intimal/medial thickness, upregulated endothelial nitric oxide synthase, and [alpha]-smooth muscle actin expression. Insulin protects autologous vein grafts possibly through the phosphatidylinositol-3 kinase/Akt signaling pathway and prevents IH in autologous vein grafts.

[Effects of insulin on cardiac function and coronary circulation in acute myocardial ischemia and reperfusion experiment with dogs].

OBJECTIVE: To study the effect of insulin on cardiac functional recovery, coronary blood flow (CBF), coronary arterial function and coronary vascular endothelial cell apoptosis following acute myocardial ischemia/reperfusion (MI/R). METHODS: In adult dogs, the left anterior descending coronary artery (LAD) was partially occluded (80% reduction in its blood flow) for 50 min and reperfused for 4 h. Vehicle (0.9% NaCl), GIK (glucose: 250 gxL(-1), insulin: 60 UxL(-1), potassium: 80 mmolxL(-1)), or GK (glucose: 250 gxL(-1), potassium: 80 mmolxL(-1)) were intravenously infused (2 mlxkg(-1)xh(-1)) 5 min before reperfusion, and CBF and left ventricular pressure were monitored. At the end of 4 h reperfusion period, coronary arteries were isolated, and the coronary vascular dysfunction, nitric oxide (NO) production and endothelial apoptosis were determined. RESULTS: During reperfusion, compared with the vehicle, GIK increased CBFLAD (19.2 ml/min +/- 2.2 ml/min) vs (14.6 ml/min +/- 1.8 ml/min) of vehicle at the end of reperfusion, P < 0.05, improved recovery of LVSP and +/- LVdP/dtmax. In vivo ischemia/reperfusion caused significant coronary vascular endothelial dysfunction as evidenced by reduced endothelium dependent vasorelaxation, decreased total NO production, and endothelial cell apoptosis as determined by TUNEL staining. Reperfusion with GIK, but not GK, markedly improved the endothelium-dependent vasorelaxation (80.3% +/- 3.8%) vs. vehicle (28.1% +/- 2.3%, P < 0.01) of coronary artery in response to ACh. GIK significantly increased total NO production (17.19 micromol/L +/- 2.18 micromol/L) versus vehicle (4.74 micromol/L +/- 2.01 micromol/L, P < 0.01) and inhibited apoptosis in coronary arterial endothelial cell (12% +/- 4%) vs vehicle (45% +/- 7%, P < 0.01). GK failed to show any significant vasculoprotection against MI/R-induced coronary vascular injury. CONCLUSION: These results demonstrate that insulin exerts cardioprotective effect by increasing CBF, reducing coronary artery injury and improving cardiac functional recovery during reperfusion, which may be partly attributable to the coronary vasculoprotective effect of insulin. The insulin-induced, NO-mediated anti-endothelial apoptotic effect may play a critical role in the insulin-induced coronary artery protective effect in MI/R.

[Application of endovascular thoracic branched aortic stent-grafts in the treatment of aortic arch dissection].

OBJECTIVE: To report the initial clinical experience of endovascular thoracic branched stent grafts in the treatment of aortic arch dissections involving the left subclavian artery. METHODS: From February 2004 to June 2004, 14 patients were cured with the endovascular thoracic branched aortic stent-grafts made by Beijing YuHengJia SciTech Co. All patients had Stanford type B aortic dissection with the entry tears just beyond the origin of the left subclavian artery by an average distance of 8.7 mm. The branched stents were consisted of the aortic section and the branched section. The diameter of the stents was 15% to 20% larger than the diameter of the landing zones of native arteries. The repair procedure was performed in angiography laboratory. The branched stent grafts were delivered under fluoroscopic guidance and implanted into the aortic arch including the left subclavian artery. RESULTS: Fourteen branched stent-grafts and 2 additional flexible stent-grafts were delivered successfully in all 14 cases. The entry tears were excluded completely, and the truth lumen of the dissection was revealed to the normal diameter in all patients. Neither peripheral complication nor death occurred. All 14 patients had recover the normal life. CONCLUSION: It demonstrates that it is possible to apply the technical feasibility of endovascular thoracic branched aortic stent graft to repair the intimal tear of dissection just beyond the left subclavian artery.

Subanesthetic isoflurane reduces zymosan-induced inflammation in murine Kupffer cells by inhibiting ROS-activated p38 MAPK/NF-κB signaling.

Volatile anesthetic isoflurane (ISO) has immunomodulatory effects. The fungal component zymosan (ZY) induces inflammation through toll-like receptor 2 or dectin-1 signaling. We investigated the molecular actions of subanesthetic (0.7%) ISO against ZY-induced inflammatory activation in murine Kupffer cells (KCs), which are known as the resident macrophages within the liver. We observed that ISO reduced ZY-induced cyclooxygenase 2 upregulation and prostaglandin E2 release, as determined by western blot and radioimmunoassay, respectively. ISO also reduced the production of tumor necrosis factor-α, interleukin-1ß, IL-6, high-mobility group box-1, macrophage inflammatory protein-1α, macrophage inflammatory protein-2, and monocyte chemoattractant protein-1 as assessed by enzyme-linked immunosorbent assays. ISO blocked the ZY-induced nuclear translocation and DNA-binding activity of nuclear factor- (NF)-κB p65. Moreover, ISO attenuated ZY-induced p38 mitogen-activated protein kinase (MAPK) activation partly by scavenging reactive oxygen species (ROS); the interregulation that ROS activated p38 MAPK followed by NF-κB activation was crucial for the ZY-induced inflammatory responses in KCs. An in vivo study by peritoneal injection of ZY into BALB/C mice confirmed the anti-inflammatory properties of 0.7% ISO against ZY in KCs. These results suggest that ISO ameliorates ZY-induced inflammatory responses in murine KCs by inhibiting the interconnected ROS/p38 MAPK/NF-κB signaling pathways.

R136K fibroblast growth factor-1 mutant induces heparin-independent migration of endothelial cells through fibrin glue.

OBJECTIVES: R136K is a mutation of fibroblast growth factor-1 (FGF-1) in which arginine replaces lysine at the primary thrombin cleavage site. This may be important in vivo in inducing endothelial cell (EC) migration and coverage of arterial injury sites by allowing R136K to be used in a fibrin glue delivery system, without thrombin-induced degradation, in the absence of heparin. The objectives of this study were to determine whether R136K, with and without heparin, can induce migration of EC and smooth muscle cells (SMC) through fibrin glue, and to compare these results with those of wild-type FGF-1; and to determine the resistance of R136K to thrombin-induced degradation versus FGF-1. METHODS: The dose-response migration through fibrin glue induced by wild-type FGF-1 and the R136K mutant in the presence and absence of heparin was tested with EC and SMC. Migration was tested with 50, 100, and 200 ng/mL of both FGF-1 and R136K, either with or without 5 U/mL of heparin. Migration of EC was also assessed after growth inhibition with mitomycin C. A novel modified Boyden chamber-type migration assay using fibrin glue on the upper surface of the chamber filter was used to test migration. The fluorescent marker calcein was used to identify those cells that had migrated through the fibrin glue and were embedded in the filter. Molecular degradation by thrombin was assessed with sodium dodecylsulfate polyacrylamide gel electrophoresis. RESULTS: For EC, R136K in the absence of heparin induced significantly more migration than did FGF-1 at 50 (P <.002), 100 (P <.0001), and 200 (P <.0001) ng/mL. In the presence of heparin, a chemotactic response of EC to cytokine was seen at all doses, with no significant difference between FGF-1 and R136K. A dose-dependent difference was noted in this group between the 100 and 200 ng/mL concentrations of cytokine (for FGF-1, P <.0001; for R136K, P <.0001). SMC showed no difference in migration with FGF-1, R136K, or negative control at any dose in the presence or absence of heparin. Gel electrophoresis demonstrated that R136K was more resistant to thrombin degradation than was FGF-1. CONCLUSION: Site-directed mutagenesis of FGF-1 to R136K enables induction of heparin-independent migration of EC through fibrin glue at an optimal concentration of 100 ng/mL. Neither FGF-1 nor R136K elicits SMC migration through fibrin glue. The ability of R136K to induce EC migration through fibrin glue in the absence of heparin may prove useful in vivo by inducing EC migration and coverage of arterial injury sites, thus potentially reducing thrombogenicity and intimal hyperplasia.