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@#Objective To compare long-term outcomes following mitral valvuloplasty (MVP) and mitral valve replacement (MVR) for native valve endocarditis (NVE). Methods Between November 1993 and August 2016, consecutive 101 patients with NVE underwent mitral surgery in our department, MVP for 52 patients and MVR for 49 patients. There were 69 males and 32 females at age of 38.1±14.9 years. The mean follow-up was 99.4±75.8 months. Results There was no statistical difference in cardiopulmonary bypass time, aortic cross-clamp time, in-hospital mortality, duration of mechanical ventilation, ICU stay or hospital stay after surgery between the two groups. Survival rate at 1, 5, 10, 20 years after surgery was 100.0%, 97.6%, 97.6%, 97.6% for MVP, and 93.5%, 84.3%, 84.3%, 66.2% for MVR with a statistical difference between the two groups (P=0.018). There was no stroke in the patients with MVP during follow-up periods. However, stroke-free survival rate at 1, 5, 10, 20 years after surgery was 100.0%, 93.9%, 89.4%, 70.2% for MVR patients with a statistical difference between the two groups (P=0.023). There was no statistical difference in recurrence of infection, perivalvular leakage and reoperation between the two groups. Composite endpoint-free survival rate at 1, 5, 10, 20 years after surgery was 100.0%, 97.6%, 92.9%, 92.9% for MVP, and 91.3%, 79.6%, 75.8%, 51.0% for MVR with a statistical difference (P=0.006). Conclusion MVP is associated with better outcomes than MVR in the patients with NVE; generalizing MVP technique in the patients with NVE is needed.
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OBJECTIVE To explore the role of gecko crude peptides (GCPs) in the proliferation, apoptosis, migration and lymphangiogenesis of human hepatocellular carcinoma cells (HepG2) and human lymphaticendothelial cells (HLECs) in vitro. METHODS The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate the anti- proliferative effect of GCPs and siRNA-VEGF-C on HepG2 cells, Hoechst 33258 staining and flow cytometry were performed to analyze cycle and apoptosis. The migration and invasion ability of cells were assayed by transwell chamber experiment and wound-healing assay. The protein and mRNA expressions of vascular endo?thelial growth factor-C (VEGF-C) and CXC chemokine receptor-4 (CXCR4) were detected by q-PCR, immunofluorescence, Western blot. The protein expressions of the extracellular signal regulated kinase (ERKI/2), c-Jun N-terminal kinase (JNK), p38-mitogen activated protein kinases (p38 MAPK), serine/threonine kinase (Akt) and phosphatidylinositol- 3- kinase (PI3K) were detected by western blot. The anti-lymphangiogenesis effect of GCPs on the HLECs was analyzed using an in vitro tube-formation assay. The protein and mRNA expressions of vascular endothelial growth factor receptor-3 (VEGFR-3) and stromal cell-derived factor-1 (SDF-1) were detected by q-PCR, Western blot. RESULTS GCPs and siRNA-VEGF-C inhibited HepG2 proliferation, invasion and migration, and the most obvious inhibitory effect was both synergistic effects. Thus, GCPs suppressed HLECs proliferation, migration and tube-like structure formationin a dose- dependent manner, and had inhibitory effect of tumor- induced lymphangiogenesis in vitro. Additionally, we found that GCPs and siRNA- VEGF- C decreased the expressions of MMP-2, MMP-9, VEGF-C, CXCR4, phospho-ERK1/2, phospho-P38, phospho-JNK and PI3K in HepG2 cells. Moreover, GCPs had a dose-dependent depressive effecton the expressions of VEGFR- 3, SDF- 1 in HLECs. CONCLUSION The low expression of VEGF- C mediated by siRNA-VEGF-C and GCPs inhibit tumor proliferation, invasion and migrationby suppressing the MAPK signaling pathway through reduced levels of VEGF-C, and GCPs inhibit tumor lymphangiogenesis by suppressing the CXCR4/SDF-1 signaling pathway through suppressed VEGF-C/VEGFR-3.
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OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma (HCC). In this study, we investigated the anti-proliferative activities and anti-angiogenesis effects of saikosaponins (SS)-b on hepatocellular carcinoma (HCC) and its regulation on VEGF/ERK/HIF-1α signal pathway. METHODS H22 hepatoma-bearing mice model and HepG-2 cells were used to study the anti-tumor and anti-angiogenesis effects of SS-b in vivo and in vitro. Pathological change of tumor tissue was observed by HE staining, the microvascular changes were detected by immunohistochemical method. The effects of SS-b on angiogenesis were examined by using the chick embryo chorioallantoic membrane (CAM) model. The effects of SS- b on proliferation, migration and invasion were investigated by MTT assay, scratch wound healing assay and transwell assay inhuman umbilical vein endothelial cell (HUVEC) and HepG2 cells in vitro. Vascular endothelial growth factor (VEGF), matrix metalloproteinase-2/9(MMP-2/9), hypoxia-inducible factor-1α (HIF-1α) expression and the phosphorylation of extracellular regulated kinase(ERK) were analyzed using RT-PCR and Western-blot. RESULTS SS-b effectively inhibited the tumor growth of H22 mice in vivo. The inhibitory rate of tumor was 49.1%, 50.7%, 66.1% in SS-b 5, 10 and 20 mg·kg-1 group respectively. HE staining results showed that SS-b induced tumor necrosis and nuclear dissolution in H22 mice. Moreover, SS-b also reduced the number of microvessels of tumor tissue in H22 mice significantly and suppressed the angiogenesis of CAM induced by b-FGF. SS-b had an obvious inhibitory effect on cell proliferation, migration and invasion of HUVEC cells and HepG-2 cells. These effects were associated with down-regulation of the expression of MMP2/9 and suppression of VEGF/ERK/HIF-1α signaling in H22 mice and Hep-G2 cells. CONCLUSION Our findings showed that SS-b exerts anti-tumor effects by inhibit?ing tumor angiogenesis via regulating VEGF/ERK/HIF-1α signal pathway in vivo and in vitro.
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OBJECTIVE To investigate the regulation of {O2 (2,4-dinitrophenyl)1-〔(4-ethoxycarbonyl) piperazin-1-yl〕diazen-1-ium-1,2-diolate}(JS-K), anitric oxide donor, on tumor energy metabolism in H22 tumor- bearing mice. METHODS The hepatoma animal model in BALB/c mice was established with H22 cell line. The JS-K group and model group were received JS-K (0.75 and 1.5 mg?kg-1) and saline via tail intravenous once every 3 d for 14 d, received 5 injections, respectively. The positive group was received 5-FU 20 mg·kg- 1 by intraperitoneal injection once a day for 14 d. On the 15th day mice were sacrificed. The tumor growth inhibition rate were calculated. The activities of hexokinase (HK), phospho?fructo kinase (PFK), pyruvate kinase (PK), succinate dehydrogenase (SDH), adenosine triphosphatase (ATPase), and the levels of lactic acid (LD) and adenosine triphosphate (ATP) in tumor tissues were de?termined by colorimetric method. RESULTS Compared with model group, the tumor mass of JS- K 0.75 and 1.5 mg·kg- 1group was significantly reduced (P<0.01),and the tumor growth inhibition rate was 23.9% and 50.3%, respectively. The activity of HK, PFK, PK, SDH and ATPase of tumor tissue in model group was (22.6±3.7, 14.4±2.6, 12.9±3.2 and 10.5±2.6)U·g-1 protein and (0.70±0.10)μmolPi·mg-1 protein per hour, respectively; which in JS-K 1.5 mg?kg-1 group was dropped by 42.0%, 26.6%, 22.7%, 23.3% and 21.7% (P<0.01, P<0.05). Compared with the model group, the level of ATP and LD in JS-K group was dropped (P<0.01). CONCLUSION JS-K can inhibit the growth of tumor in H22 tumor-bearing mice and its mechanism may be related to regulating the tumor energy metabolism with inhibition of glycolysis and aerobic oxidation.
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OBJECTIVE Metastasis-associated in colon cancer-1 (MACC1) is an oncogene that has been newly identified. It promotes tumor proliferation and invasion via the MET pathway. Our study investigated the effects of Saikosaponin-b(SS-b) on the proliferation and apoptosis of HepG2 cells and its regulation on MACC1/c-Met/Akt signaling pathway. METHODS HepG2 cells were treated with SS-b (10-800 g·L-1) for 48 h in vitro. The CCK-8 assay was used to assess cell proliferation, and cell apoptosis was determined by Hoechst33258 staining, AnnexinⅤ/PI staining and caspase 3 assay. RT-PCR was used to examine the expression of MACC1, c- MET and hepatocyte growth factor (HGF) mRNA. MACC1 protein was detected by Western blot and immunohistochemistry. The protein expressions of p-c-MET, c-MET, p-AKT, AKT, p-BAD, BAD were measured by Western blot. RESULTS SS-b inhibited the growth of HepG2 cells in dose-dependent way and induced cell apoptosis significantly. HepG2 cells showed karyopyknosis, fragmentation and fluorescence highlight in SS-b treatment group. FCM results showed that apoptosis rate of HepG2 cells increased with SS- b concentration. The immunofluores?cence results showed that the MACC1 expression decreased significantly in HepG2 cells treated with SS-b. The expression levels of MACC1, c-MET and HGF mRNA in HepG2 cells were significantly inhibited by SS-b. SS-b also significantly decreased the protein expressions of MACC1, p-c-MET and p-AKT while increased the expression of p-BAD and caspase 3 in HepG2 cells(P<0.05). CONCLUSION SS-b inhibited the proliferation and induced the apoptosis of HepG2 cells by targeting the MACC1/c-Met/Akt signaling pathway.
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OBJECTIVE In order to investigate the possible anti-tumor molecular mechanisms of gecko polypeptide mixture (GPM). METHODS RNA-seq technology was used to identify the differen?tially expressed genes of human hepatocellular carcinoma (HCC) HepG2 cells treated with or without GPM. The HepG2 cells were treated with different concentration of GPM (0, 0.1, 0.2, 0.3, 0.4 mg·mL-1) for 6 h, 12 h and 24 h, respectively. MTT assay was used to detect the viability of HepG2 cells. DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells. Western blot analysis was applied to observe the expression of apoptosis-related proteins in HepG2 cells. RESULTS The results showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner. We applied many analysis methods, including differen?tially expressed genes analysis, Gene Ontology (GO) enrichment analysis, KEGG pathway enrichment analysis, protein- protein interaction network analysis to screen out possible molecular mechanisms. ER-nucleus signaling pathway, cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM. GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway. The mechanism is closely related to ERs, which might be beneficial for clinical therapy of HCC. CONCLUSION GPM can inhibit cells proliferation and induce apoptosis in HepG2 cells. The gene expression profile of GPM in HepG2 cells was obtained. The present study revealed the potential anti-tumor mechanism of GPM.
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Complementary DNA (cDNA) is valuable for investigating protein structure and function in the research of life science, but it is difficult to obtain by traditional reverse transcription. In this study, we employed a novel strategy to clone the human leukemia inhibitory factor (hLIF) gene cDNA from genomic DNA directly isolated from the mucous membrane of mouth. The hLIF sequence can be acquired within a few hours by means of amplification of each exon and splicing using overlap-PCR. Thus, the new approach developed in this study is simple, time- and cost-effective, and it is not limited to particular gene expression levels of each tissue.