miR-632 Promotes Laryngeal Carcinoma Cell Proliferation, Migration, and Invasion Through Negative Regulation of GSK3ß.

Laryngeal cancer, one of the most common head and neck malignancies, is an aggressive neoplasm. Increasing evidence has demonstrated that microRNAs (miRNAs) exert important roles in oncogenesis and progression of diverse types of human cancers. miR-632, a tumor-related miRNA, has been reported to be dysregulated and implicated in human malignancies; however, its biological role in laryngeal carcinoma remains to be elucidated. The present study aimed at exploring the role of miR-632 in laryngeal cancer and clarifying the potential molecular mechanisms involved. In the current study, miR-632 was found to be significantly upregulated both in laryngeal cancer tissues and laryngeal cancer cell lines. Functional studies demonstrated that miR-632 accelerated cell proliferation and colony formation, facilitated cell migration and invasion, and enhanced the expression of cell proliferation-associated proteins, cyclin D1 and c-myc. Notably, miR-632 could directly bind to the 3'-untranslated region (3'-UTR) of glycogen synthase kinase 3ß (GSK3ß) to suppress its expression in laryngeal cancer cells. Mechanical studies revealed that miR-632 promoted laryngeal cancer cell proliferation, migration, and invasion through negative modulation of GSK3ß. Pearson's correlation analysis revealed that miR-632 expression was inversely correlated with GSK3ß mRNA expression in laryngeal cancer tissues. Taken together, our findings suggest that miR-632 functions as an oncogene in laryngeal cancer and may be used as a novel therapeutic target for laryngeal cancer.

Anti-inflammatory activity of resveratrol prevents inflammation by inhibiting NF­κB in animal models of acute pharyngitis.

Recent studies have demonstrated that resveratrol can reduce blood sugar, improve insulin resistance, regulate abnormalities in lipid metabolism, and lower the secretion and expression of inflammatory factors. The present study investigated the anti­inflammatory effects of resveratrol in animal models of acute pharyngitis, and its possible mechanisms. Commercial ELISA kits were used to measure tumor necrosis factor­α, interleukin (IL)­6, macrophage inflammatory protein­2, cyclooxygenase­2 levels and caspase­3/9 activity. Toll­like receptor (TLR)­4, myeloid differentiation primary response protein MyD88, phosphorylated (p)­nuclear factor (NF)­κB and p­IκB were analyzed using western blotting. In a rabbit model of acute pharyngitis, it was demonstrated that resveratrol inhibited tumor necrosis factor­α and interleukin­6 serum levels, macrophage inflammatory protein­2 and cyclooxygenase­2 activity levels, reactive oxygen species production and caspase­3/9 activity. Resveratrol suppressed NACHT, LRR and PYD domains­containing protein 3 and caspase­1 protein expression, and reduced IL­1ß and IL­18 protein expression in animal models of acute pharyngitis. Additionally, resveratrol suppressed TLR4 and myeloid differentiation primary response protein 88 protein expression, and reduced p­NF­κB and increased p­IκB protein expression in animal models of acute pharyngitis. In conclusion, these findings indicated that the anti­inflammatory activity of resveratrol prevents acute pharyngitis­induced inflammation by inhibiting NF­κB in animal models. Therefore, these data suggested an important clinical application of resveratrol in preventing acute pharyngitis.

Effects of dietary oregano essential oil and vitamin E supplementation on meat quality, stress response and intestinal morphology in pigs following transport stress.

This study investigates the effects of dietary oregano essential oil (OEO) and vitamin E (Vit E) supplementation on meat quality, stress response and intestinal morphology in pigs following transport stress. A total of 288 finishing pigs were randomly assigned to three groups: a basal diet or a basal diet supplemented either with 200 mg/kg Vit E or 25 mg/kg OEO. After a 28-day feeding trial, total of 132 finishing pigs according diet and transport stress were assigned to one of four treatment groups: 1) control treatment without transport stress (Control group), 2) control treatment with 5-hr transport stress (Negative group), 3) Vit E treatment with 5-hr transport stress and 4) OEO treatment with 5-hr transport stress. Transport stress pigs had lower muscle 45 min pH (pHi) and higher drip loss than control pigs. Dietary OEO and Vit E supplementation significantly increased 45min pH under transport stress, and the OEO groups produced lower 24-hr drip loss values (P<0.05) than that of pigs from the negative group. The OEO-supplemented pigs showed decreased serum levels of creatine kinase (CK) and cortisol (P<0.05), and decreased Hsp 27 (heat shock protein 27) and Hsp 70 (heat shock protein 70) mRNA expression in the muscle (P<0.05). Additionally, histological analysis revealed intestinal epithelial damage in transport stress pigs that was reversed by dietary supplementation with OEO. In conclusion, supplementation with dietary OEO may be superior to supplementation with dietary Vit E in alleviating the meat quality, stress response and intestinal morphology of pigs after challenge due to transportation stress.

[Hybrid operation for acute left leg deep venous thrombosis secondary to left iliac vein compression syndrome: analysis of 36 cases].

OBJECTIVE: To evaluate the surgical techniques for acute left deep venous thrombosis (LDVT) secondary to left iliac vein compression syndrome (IVCS). METHODS: Thirty-six patients with acute LDVT secondary to IVCS received inferior vena cava filter placement, and in 2 of the cases, stent implantation was canceled for acute episode of obsolete DVT. The remaining 34 patients underwent left femoral venotomy for iliofemoral thrombectomy with Fogarty catheter and distal femoral vein thrombus removal by sequential compression of the legs, followed by implantation of stent-graft (2 cases) or bare-metal stents (32 cases) in the left common iliac veins. With routine anticoagulation and thrombolytic treatments, the patients were regularly examined for postoperative blood flow in the affected limb. RESULTS: In 2 of the cases undergoing bare-metal stent implantation, the residue thrombi were squeezed into the stent by balloon, which was managed subsequently with local thrombolysis. One patient with bare-metal stent implantation received a secondary stenting for posterior stent displacement. Three patients had self-limited bleeding due to decreased serum FBG. Significant improvements were achieved at 3, 6, 30 and 180 days postoperatively in the circumferences of the affected limb (P<0.05) and in the levels of D-dimer (P=0.011), and FBG level showed no significant variations (F=1.163, P=0.345). The total rate of excellent outcomes was 83.3% (26/34) with a total effective rate of 91.2% (31/34) in these cases. CONCLUSIONS: Thrombectomy to revascularize the inflow tract and stent implantation to enlarge stenosed iliac veins are key issues in treatment of acute LDVT secondary to IVCS.

Screening of Ganoderma strains with high polysaccharides and ganoderic acid contents and optimization of the fermentation medium by statistical methods.

Polysaccharides and ganoderic acids (GAs) are the major bioactive constituents of Ganoderma species. However, the commercialization of their production was limited by low yield in the submerged culture of Ganoderma despite improvement made in recent years. In this work, twelve Ganoderma strains were screened to efficiently produce polysaccharides and GAs, and Ganoderma lucidum 5.26 (GL 5.26) that had been never reported in fermentation process was found to be most efficient among the tested stains. Then, the fermentation medium was optimized for GL 5.26 by statistical method. Firstly, glucose and yeast extract were found to be the optimum carbon source and nitrogen source according to the single-factor tests. Ferric sulfate was found to have significant effect on GL 5.26 biomass production according to the results of Plackett-Burman design. The concentrations of glucose, yeast extract and ferric sulfate were further optimized by response surface methodology. The optimum medium composition was 55 g/L of glucose, 14 g/L of yeast extract, 0.3 g/L of ferric acid, with other medium components unchanged. The optimized medium was testified in the 10-L bioreactor, and the production of biomass, IPS, total GAs and GA-T enhanced by 85, 27, 49 and 93 %, respectively, compared to the initial medium. The fermentation process was scaled up to 300-L bioreactor; it showed good IPS (3.6 g/L) and GAs (670 mg/L) production. The biomass was 23.9 g/L in 300-L bioreactor, which was the highest biomass production in pilot scale. According to this study, the strain GL 5.26 showed good fermentation property by optimizing the medium. It might be a candidate industrial strain by further process optimization and scale-up study.

[Efficacy of regional administration of urokinase and argatroban via small saphenous vein catheter for treatment of acute deep venous thrombosis in the lower limb].

OBJECTIVE: To investigate the clinical value of local regional administration of urokinase and argatroban through small saphenous vein (SSV) catheter in the treatment of acute deep venous thrombosis in the lower limb (LDVT). METHODS: Fifty-six patients with acute LDVT were prospectively randomized into the study group (21 cases, 24 limbs) and control group (35 cases, 36 limbs) for treatment with urokinase and argatroban regionally administered via the SSV catheter and with the same agents given via the peripheral vein, respectively. The patients were examined for changes in serum fibrinogen (FBG) and D-dimer and the perimeter of the affected limbs, and the complications in relation to the agents were observed. RESULTS: By corrected Chi-square test, the incidence of complications was significantly lower in the study group than in the control group (1/21 vs 4/36, χ(2)=1.92, P≤0.05). Wilcoxon's sign rank test suggested no statistically significant difference between the two groups in the total effective rate (95.8% vs 94.4%, V=0.52, P>0.05), but the total excellent rate differed significantly between them (83.3% vs 55.6%, V=2.36, P≤0.05). Serum FBG underwent no significant variations in the study group during thrombolysis (P>0.05), but decreased significantly in the control group (P≤0.05). The decreases in serum D-dimer and perimeter of the affected limbs occurred earlier in the study group than in the control group (P≤0.05). CONCLUSION: Regional administration of urokinase and argatroban via small saphenous vein catheter can promote the thrombolytic effect and reduce the risk of hemorrhage in the treatment of LDVT.

[Experience with endovenous laser treatment combined with high ligation and Muller's phlebectomy for C3-6 grade primary superficial varicose in the lower limbs].

OBJECTIVE: To summarize the experience with endovenous laser treatment(EVLT) combined with high ligation and Muller's phlebectomy for primary superficial varicose in the lower limbs. METHODS: In 95 patients with C3-6 grade primary superficial varicose in 146 lower limbs, the extent of varicose was accurately marked, the guiding wires were manipulated precisely, and the proximal great saphenous veins (GSV) were ligated after exsanguinations. The stems of the GSV were ablated with laser with the lower limbs lift up and pressed hard along the stems, and Muller's incisions were carefully planned. RESULTS: All the operations were completed successfully. The guiding wires entered into the deep veins through the communicating branches in 2 limbs, 1 patient experienced capillary hemorrhage from Muller's incisions, 8 had thrombotic phlebitis of the GSV, 7 sustained heat-related injury of the saphenous nerves, 1 experienced skin heat-related lesion, 2 developed hematoma in the inguinal region, 2 had pitting edema in the dorsum of the foot, 1 had fat liquefaction of the Muller incision, and 1 showed rejection of the thrum. After conservative treatment, all the patients recovered and were discharged. Part of the superficial varicose remained after the operation in 6 limbs. CONCLUSION: It is necessary to standardize the routine procedure of EVLT combined with high ligation and Muller's phlebectomy to reduce the complications.

[Expression of survivin mRNA and protein in mitomycin-treated hepatoma carcinoma Hepa1-6 cells].

OBJECTIVE: To investigate survivin mRNA and protein expressions in mitomycin (MMC)-treated hepatoma carcinoma Hepa1-6 cells in vitro. METHODS: Hepa1-6 cells were cultured in vitro in the presence of MMC at the concentrations of 1.0, 3.0 and 9.0 microg/ml, respectively, and 1 day and 3 days after the culture, the cell growth inhibition was assessed using MTT assay and the expressions of survivin were detected by RT-PCR and Western blotting. RESULTS: MMC at the concentration of 9.0 microg/ml resulted in significantly greater growth inhibition of the Hepa-6 cells than MMC at 1.0 and 3.0 microg/ml, and at the latter two concentrations, MMC treatment for 3 days did not produce obvious cell growth inhibition. Survivin expressions at both the mRNA and protein levels in Hepa1-6 cells were significantly decreased 1 day after MMC treatment at the 3 concentrations, and after 3-day MMC treatment at 1.0 and 3 microg/ml, survivin expressions increased to exceed the control level, whereas survivin maintained the low expression levels in cells treated with 9 microg/ml MMC for 3 days. CONCLUSION: Survivin expression in Hepa1-6 cells increases in response to MMC treatment at low doses, which might be one of the reasons for chemotherapeutic drug resistance.

[Protective effects of the induction of heme oxygenase-1 on ischemia reperfusion lung injury: in vivo experiment with rats].

OBJECTIVE: To investigate the protective effects of the induction of heme oxygenase-1 (HO-1) on ischemia/reperfusion lung injury. METHODS: Forty Sprague-Dawley rats were randomly divided into four equal groups: sham group, lung ischemia/reperfusion injury (I/R) group, undergoing ligaturing of the left lung hilum for 30 minutes followed by reperfusion for 120 minutes; hemin group, undergoing intraperitoneal injection of hemin, an inducer of HO-1, 48 hours before the ligation and reperfusion; zinc protoporphyrin (ZnPP) group, undergoing intravenous injection of ZnPP, an inhibitor of heme oxygenase, 15 min after the ischemia-reperfusion; and sham operation group, undergoing sham operation. Two hours after the I/R arterial blood samples were collected and then the left lungs of the rats were taken out. Plasma tumor necrosis factor-alpha (TNF-alpha) and lung superoxide dismutase (SOD) activity were examined. Lung wet-to-dry weight (W/D) ratio was measured. The ultrastructure of the pulmonary alveoli and its capillaries were studied by using transmissional electronmicroscopy. RESULTS: The lung W/D ratio of the hemin group was 5.92 +/- 0.66, significantly lower than that of the I/R group (7.55 +/- 0.66, P < 0.01), and that of the ZnPP group (7.34 +/- 0.39, P < 0.01). The SOD activity of the hemin group was 6.5 +/- 0.6 U/mg protein, significantly higher than those of the I/R group and ZnPP group (2.8 +/- 0.4 U/mg protein and 3.0 +/- 0.4 U/mg protein respectively, both P < 0.01). The plasma TNF-alpha was 180.36 +/- 12.46, significantly lower than those of the I/R and ZnPP groups (452.26 +/- 22.59 and 438.59 +/- 30.26 respectively, both P < 0.01). Transmissional electronmicroscopy showed that the microscopic structure of the sham group was normal and that the pathological changes of hemin group were milder then those of the T/R and ZnPP groups. CONCLUSION: The induction of heme oxygenase-1 can protect effectively the lesion of lung pathology in ischemia reperfusion in vivo.

[Arginine supplementation in total parenteral nutrition improves postoperative cellular immune function of patients with hepatocellular carcinoma after operation].

OBJECTIVE: To explore the effect of total parenteral nutrition (TPN) supplemented with arginine on cellular immune function of patients with hepatocellular carcinoma (HCC) after radical tumor resection. METHODS: Fifty-six HCC patients undergoing radical surgery received fat-free TPN support, routine TPN or TPN with arginine supplementation, and their clinical data were analyzed prospectively. The percentages of T lymphocyte subpopulation and national killer (NK) cells in peripheral blood are determined, and the levels of interleukin-2 (IL-2), IL-4 and interferon-gamma (IFN-gamma) were measured. RESULTS: No marked changes were noted in peripheral blood CD4+, CD8+ T cells and NK cells, or in IL-2, IL-4 and IFN-gamma levels after fat-free TPN and routine TPN support. TPN supplemented with arginine resulted in significant increase in CD4+ T cells, NK cells and CD4+/CD8+ T cell ratio in the peripheral blood, as well as in IL-2 and IFN-gamma levels. Peripheral blood IL-4 level was decreased significantly. CONCLUSION: TPN with arginine supplementation can augment the percentages of CD4+ T lymphocytes and NK cells, and increase IL-2 and IFN-gamma levels, suggesting that arginine can enhance cell-mediated immunity in postoperative patients with HCC.

[Microwave coagulation at different temperatures for hepatocellular carcinoma management: efficacy evaluation by enzyme histochemical staining].

OBJECTIVE: To compare the application of HE and enzyme histochemical staining in assessing the viability of hepatocellular carcinoma (HCC) cells coagulated by microwave ablation at different temperatures. METHODS: Two groups of mice (n=6) with transplanted homogenic HCC were treated by microwave ablation at 60 degrees C and 50 degrees C for 3 min, respectively. Before and after microwave ablation, paraffin sections and frozen sections of the tumors were prepared for routine HE staining and enzyme histochemical staining with nicotinamide adenine dinucleotide diaphorase (NADH-diaphorase), respectively, and observed under microscope. RESULTS: Shortly after microwave ablation, the morphology and arrangements of the nucleus of the ablated tumor cells in the two groups showed no obvious alteration in HE stained sections, but in sections with enzyme histochemical staining, the activity of NADH-diaphorase in ablated tumor tissue at 60 degrees C disappeared, suggesting the death of HCC cells; sporadic activity of the enzyme was detected in the coagulated tumor at 50 degrees C, indicating tumor cells surviving the ablation. The ablation effect was markedly different between the two groups (P<0.01). CONCLUSION: HE staining is not suitable for evaluation of HCC destruction immediately after microwave ablation, and detection of NADH-diaphorase activity with the enzyme histochemical method better suits this purpose.

Effects of endothelin-1 on differentiation of cardiac myocyte induced from rabbit bone marrow stromal cells.

BACKGROUND: Cardiomyocyte transplantation for the therapy of myocardial ischaemia is being paid close attention. However, how the microenvironment controls the differentiation of transplanted bone marrow stromal cells (BMSCs) is unknown. Endothelin-1 (ET-1), a cytokine, increases during myocardial infarction, but it is not known whether ET-1 is responsible for the fate of transplanted BMSCs. In the present study, we investigated the effects of ET-1 on differentiation and maturation of induced rabbit BMSCs, in vitro, to elucidate the cellular biological mechanisms. METHODS: The proliferation of BMSCs isolated from femur of rabbits was induced by ET-1 only, by 5-azacytidine (5-aza) or ET-1 combined with 5-aza. After 4 weeks of induced culturing, the differentiation rate and the diameter of induced myocyte like cells were estimated and the expressions of GATA-4 protein and phosphorylation level were assayed by Western-blot, RT-PCR analysis of beta-myosin heavy chain (MHC). mRNA expression, levels of troponin-I by immunohistochemical staining and ultrastructure of induce-cultured BMSCs were also determined. RESULTS: By induction with ET-1 and 5-aza, mean cell diameter of induced BMSCs was larger than induced with 5-aza [(6.26 +/- 0.22) microm cf (5.29 +/- 0.19) microm] (P < 0.001). There was no difference in rate of differentiation of myocyte like cells between the groups induced with 5-aza and ET-1 combined with 5-aza [(29.82 +/- 0.23)% cf (29.94 +/- 0.18)%] (P > 0.05). The expressions of GATA-4 protein and phosphorylation were enhanced significantly in groups induced with ET-1 combined with 5-aza (P < 0.05). In the group induced with ET-1 combined with 5-aza, expression of beta-MHC mRNA was higher than control [(0.122 +/- 0.008) cf (0.022 +/- 0.003)] (P < 0.01), and more troponin-I positive cells were also detected in this group. Differentiated BMSCs showed formations of myofilaments and primitive sarcomere, i.e., morphological characteristics of myocyte like cells. CONCLUSIONS: This study suggests that induced culturing of BMSCs by ET-1 combined with 5-aza can express cardiomyocytic characteristics whereas ET-1 alone could not induce BMSCs to differentiate to myocyte like cells. ET-1 upregulates the expression of GATA-4 protein and phosphorylation level of induced BMSCs, and rapidly promotes the differentiation and maturation of myocyte like cells from BMSCs.