MiR-204 reduces apoptosis in rats with myocardial infarction by targeting SIRT1/p53 signaling pathway.

OBJECTIVE: The aim of this study was to investigate the influence of micro ribonucleic acid (miR)-204 on rats with myocardial infarction by targeting the silent information regulator 1 (SIRT1)/p53 signaling pathway. MATERIALS AND METHODS: A total of 36 Sprague-Dawley rats were randomly divided into three groups, including: sham-operation group (n=12), model group (n=12) and miR-204 mimics group (n=12). The rats in the sham-operation group only underwent thoracotomy, without myocardial infarction injury. Meanwhile, the rats in model group and miR-204 mimics group were utilized to establish the models of myocardial infarction, and then, intervened with normal saline and miR-204 mimics, respectively. The morphology of myocardial tissues was observed via hematoxylin-eosin (HE) staining. Immunofluorescence was performed to detect the expression of Caspase-3. Target genes of miR-204 were analyzed using bioanalysis software. Western blotting (WB) assay was applied to measure the relative protein expression of SIRT1. MiR-204 expression and the messenger RNA (mRNA) expressions of SIRT1 and p53 were measured via quantitative Polymerase Chain Reaction (qPCR). Furthermore, cell apoptosis was determined through terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. RESULTS: HE staining showed that the morphology of myocardial tissues was normal in sham-operation group. Severe myocardial tissue injury was visible in model group, and the injury was relieved in miR-204 mimics group when compared with model group. The results manifested that the positive expression of Caspase-3 in cardiac tissues increased remarkably in the model group and miR-204 mimics group in comparison with sham-operation group (p<0.05). Meanwhile, it was evidently lower in miR-204 mimics group than model group (p<0.05). Based on the analysis via bioanalysis software, SIRT1 was the target gene of miR-204. WB results revealed that the relative protein expression level of SIRT1 was elevated notably in the other two groups compared with the 2sham-operation group (p<0.05). However, it was markedly lowered in miR-204 mimics group in contrast with model group (p<0.05). QRT-PCR results demonstrated that the model group and miR-204 mimics group exhibited distinctly lower expression of miR-204 but higher mRNA expressions of SIRT1 and p53 than sham-operation group (p<0.05). However, miR-204 mimics group exhibited prominently higher expression of miR-204 but lower mRNA expressions of SIRT1 and p53 than model group (p<0.05). Finally, the results of TUNEL assay demonstrated that the apoptosis rate increased remarkably in the model group and miR-204 mimics group when compared with sham-operation group (p<0.05). However, it decreased notably in miR-204 mimics group in comparison with model group (p<0.05). CONCLUSIONS: MiR-204 reduces the apoptosis level in rats with myocardial infarction via targeted inhibition of the SIRT1/p53 signaling pathway.

Suberoylanilide hydroxamic acid attenuates epidural fibrosis via inhibiting myofibroblast differentiation and increasing fibroblast apoptosis.

OBJECTIVE: Epidural fibrosis represents a fatal stage of failed back surgery syndrome (FBSS) of known and idiopathic etiology, but no valid therapy is presently available. Previous evidence demonstrated that suberoylanilide hydroxamic acid (SAHA), a histone deacetylases inhibitor, has antifibrotic and anti-inflammatory potential. Current studies have proved that SAHA inhibits myofibroblast differentiation and increases fibroblast apoptosis to attenuate epidural fibrosis. The purpose of this study was to investigate the effect and mechanism of SAHA on repressing epidural fibrosis. PATIENTS AND METHODS: First, the levels of acetylation of histone and α-tubulin in adult human fibroblasts (AHF) and human epidural fibroblasts (HEF) were analyzed following SAHA and transforming growth factor-ß(TGF-ß) treatment. Then, mRNA and protein obtained from human fibroblasts following TGF-ß activation and SAHA treatment in vitro culture were used to test the influence of SAHA on the activation and apoptosis of fibroblasts, so as to further explore the related mechanism of SAHA. Then, a laminectomy model was established in rats to observe the therapeutic effect of SAHA on epidural scar tissue. RESULTS: The present research proved that the increases of HDAC 3 and α-tubulin were observed in AHF and HEF after TGF-ß administration, but SAHA decreased HDAC 3 and α-tubulin expressions. In addition, cell study demonstrated that SAHA inhibited fibroblast activation via decreasing TGF-ß function and accelerated apoptosis by promoting cleaved-caspase-3. In the epidural fibrosis model, it was found that SAHA weakened scar hyperplasia and collagen deposition, and effectively inhibited the process of epidural fibrosis. CONCLUSIONS: These results indicated that SAHA inhibited HDAC 3 expression, decreased TGF-ß effect, and enhanced caspase-3 in fibroblasts, leading reduction of myofibroblast activation and apoptosis elevation. Hence, SAHA ameliorated epidural fibrosis development.

MicroRNA-370 suppresses SOX12 transcription and acts as a tumor suppressor in bladder cancer.

OBJECTIVE: Dysregulation of microRNA-370 (miR-370) is involved in a variety of cancers, but its roles in bladder cancer (BC) remain largely unexplored. Therefore, we designed this study to explore the role of miR-370 in BC. PATIENTS AND METHODS: We took advantage of biochemical assays, including RT-qPCR, Western blot, CCK-8, flow cytometry, transwell, xenograft tumor formation, and immunohistochemistry (IHC) for research. RESULTS: The expression of miR-370 was found to be downregulated during the development of BC, highly correlating with the malignant transformation of tumors. The overexpression of miR-370 led to enhanced apoptosis in BC cells, while inhibiting cell proliferation, migration, and invasion, effectively blocking cancer metastasis. Additionally, we identified SOX12, a known human oncogene, as a direct target of miR-370, showing that upregulation of SOX12 attenuated miR-370-mediated tumor suppression, promoted tumor growth, and epithelial-mesenchymal transition (EMT) in BC. CONCLUSIONS: Taken together, these findings help to elucidate the roles of miR-370 as a tumor suppressor in BC, providing a potential target for diagnosis and treatment of BC.

Semaphorin-7A contributes to growth, migration and invasion of oral tongue squamous cell carcinoma through TGF-ß-mediated EMT signaling pathway.

OBJECTIVE: Oral tongue squamous cell carcinoma (OTSCC) is the most frequently encountered malignant epithelial tumors. Semaphorin-7A is a membrane-associated/secreted protein that plays an essential role in the migration and progression of human malignancies. We aimed to investigate the mechanisms of Semaphorin-7A in the growth and migration of OTSCC. MATERIALS AND METHODS: The expressions of Semaphorin-7A in cells were tested by RT-PCR, Western blot, and Immunofluorescence, separately. The activities of OTSCC cells (HSC-3 and Tca8113) were analyzed by MTT, following treatment with Semaphorin-7A or PBS. The migration, invasion, and apoptosis of cells were also determined. The protein expressions of epithelial mesenchymal transition (EMT) pathway were analyzed by Western blot, after treated with Semaphorin-7A in vitro and in vivo. Finally, the mouse model of OTSCC was treated with antibody target for Semaphorin-7A (AntiSema-7A), Semaphorin-7A or PBS, then the tumor size was determined, and histopathological examination and western blot was applied for further confirmation. RESULTS: In OTSCC cells, Semaphorin-7A was highly expressed, and Semaphorin-7A promoted growth in multiple metastatic OTSCC cell lines. Further study indicated that Semaphorin-7A resulted in up-regulation of Snail, N-cadherin and Vimentin expression, and downregulating of E-cadherin. In addition, The Ets2-repressor factor (ERF) expression was down-regulated, and transforming growth factor (TGF-ß)-induced EMT was promoted in OTSCC cells. Then, the proteins of collagen types I (CT-I) and fibronectin (FIB) were also up-regulated after Semaphorin-7A treatment. Furthermore, our results indicated that inhibition of Semaphorin-7A by antibody target for Semaphorin-7A (AntiSema-7A) suppressed OTSCC growth and increased survival in a mouse model of OTSCC. Histopathological examination confirmed the inhibitory effects in vivo. CONCLUSIONS: Semaphorin-7A promoted growth and migration of OTSCC by regulating TGF-ß-induced EMT signaling pathway in OTSCC cells, which provided a new interconnection between the Semaphorin-7A and TGF-ß-induced EMT signaling pathway.

Left circumflex coronary-to-pulmonary artery fistula as the exclusive collateral to the occluded left anterior descending artery.

A 64 year-old male presented with a five month history of effort angina. Non-invasive studies demonstrated preserved left ventricular function and a modest stress-induced myocardial perfusion defect at the anterior wall. Coronary angiography revealed occlusion of the proximal left anterior descending coronary artery with its distal segment well supplied by collaterals branching from a left circumflex-to-main pulmonary artery fistula. The occluded left anterior descending coronary artery was recanalised by percutaneous interventions, the collaterals vanished immediately, and the patient lived free of symptoms for the following five months.

Right axillosubclavian vein thrombosis resolved by percutaneous rheolytic thrombectomy in a patient with a recurrent gastrointestinal stromal tumour undergoing imatinib mesylate treatment.

The manifestation and management of spontaneous axillosubclavian vein thrombosis in patients with gastrointestinal stromal tumour (GIST) undergoing oral chemotherapy have never been described. We report a patient with a recurrent GIST who was receiving maintenance imatinib yet developed a right axillosubclavian vein thrombotic occlusion. The occluded vein was unresponsive to systemic anticoagulation but was reopened by percutaneous rheolytic thrombectomy and has shown good long-term patency. Thus, for patients with recurrent GIST undergoing imatinib therapy, axillosubclavian vein thrombosis might manifest as a complication and could be managed with rheolytic thrombectomy, which thoroughly removes intravascular thrombus and effectively re-vascularizes the thrombosed vessel uneventfully.

Sam68 expression and cytoplasmic localization is correlated with lymph node metastasis as well as prognosis in patients with early-stage cervical cancer.

BACKGROUND: This study was aimed at investigating the role and molecular mechanism of Sam68 in cervical cancer lymph node metastasis. MATERIALS AND METHODS: Sam68 expression profile was detected by quantitative polymerase chain reaction, western blotting and immunohistochemical staining. Short hairpin RNA interfering approach was employed to suppress endogenous Sam68 expression in cervical cancer cells to determine its role in metastasis and the possible mechanism. RESULTS: Sam68 expression in cervical cancer was significantly up-regulated at both messenger RNA and protein levels compared with that in normal cervical tissues. The high expression level of Sam68 and its cytoplasmic localization were significantly associated with risk factors including pelvic lymph node metastasis (P < 0.001), and served as independent prognostic factors for predicting shortening of the overall survival time and disease-free survival time in patients with early-stage cervical cancer. Moreover, down-regulation of Sam68 in cervical cancer cells remarkably inhibited cellular motility and invasion. In addition, down-regulation of Sam68 reversed epithelial-mesenchymal transition through inhibiting the Akt/ GSK-3ß/Snail pathway. CONCLUSION: This study demonstrated that Sam68 could induce cervical cancer lymph node metastasis through regulating epithelial-mesenchymal transition, and Sam68 expression profile possessed the potential to serve as predictors of pelvic lymph node metastasis in cervical cancer patients.

Late proximal coronary artery stenosis complicating percutaneous endovascular catheterisation procedures.

BACKGROUND: Late-onset proximal coronary artery stenosis caused by preceding percutaneous catheterisation procedures remains under-surveyed. METHODS: From 1993, all patients undergoing percutaneous coronary procedures and a second session within 3 years were included except those ever treated by coronary bypass surgery or chest radiotherapy during this 3-year period. Emergence of a new lesion or worsening of an initially insignificant lesion to >50% of diameter stenosis at the never-treated ostial/proximal coronary segment on the follow-up angiogram was defined as late coronary stenosis caused by the previous catheterisation procedure and was analysed. RESULTS: From January 1993 to December 2005, 3240 patients who underwent 5025 procedures met the inclusion criteria. Of them, 23 patients experienced an event of late coronary artery stenosis (overall incidence 0.46%), and interventional procedures, specifically shaped catheters (Voda, XB, Amplatz Left) and atherosclerosis vulnerability correlated with risks of adverse events. Most of these events could be managed by contemporary medical, interventional, or surgical strategies, yet hazards of mortality and long-term restenosis still existed from this catheter-induced complication. CONCLUSIONS: Percutaneous catheterisation procedures could be complicated by late proximal coronary artery stenosis. Thus, when conducting these procedures, operators should select and manipulate catheters with caution, especially in patients with susceptible clinical characteristics.

Impact of chronic advanced aortic regurgitation on the perioperative outcome of noncardiac surgery.

BACKGROUND: Whether and how chronic advanced aortic regurgitation (AR) impacts the perioperative outcome of noncardiac surgery remains unclear. METHODS: From November 1999 to December 2006, all patients undergoing noncardiac operations and ever examined by echocardiography within the last 6 months were screened. Those with chronic moderate-severe or severe AR were enrolled, provided they were not already trachea-intubated or aortic valve operated, and the surgery was not performed under local anesthesia. Case-matched subjects without significant AR served as controls. The perioperative outcomes of these patients were analyzed, and independent prognostic correlates were investigated by multivariate logistic regression analysis. RESULTS: A total of 167 patients (male 131, mean age of 75 years) complying with the enrollment criteria were studied. Compared with the other 167 case-matched control peers, patients with advanced AR risked potential hazards of serious hemodynamic instability (0.6%) and circulatory collapse (1.2%) during surgery despite the similar incidence of overall cardiac adverse events, and were further distressed with more cardiopulmonary complications (16.2% vs. 5.4%, P=0.003) and in-hospital deaths (9% vs. 1.8%, P=0.008) post-operatively. Multivariate regression analysis confirmed the correlation of advanced AR with perioperative mortality, and identified depressed left ventricular function, renal dysfunction, high surgical risk, and lack of cardiac medication as predictors of in-hospital death. CONCLUSION: Chronic advanced AR complicates the perioperative outcome of noncardiac surgery as reflected by frequent cardiopulmonary morbidities and in-hospital deaths, especially when coexisting with specified high-risk clinical and surgical characteristics.

A standardized extract of Ginkgo biloba suppresses doxorubicin-induced oxidative stress and p53-mediated mitochondrial apoptosis in rat testes.

BACKGROUND AND PURPOSE: Doxorubicin evokes oxidative stress and precipitates cell apoptosis in testicular tissues. The aim of this study was to investigate whether the Ginkgo biloba extract 761 (EGb), a widely used herbal medicine with potent anti-oxidant and anti-apoptotic properties, could protect testes from such doxorubicin injury. EXPERIMENTAL APPROACH: Sprague-Dawley male rats (8 weeks old) were given vehicle, doxorubicin alone (3 mg kg(-1) every 2 days for three doses), EGb alone (5 mg kg(-1) every 2 days for three doses), or EGb followed by doxorubicin (each dose administered 1 day after EGb). At 7 days after the first drug treatment oxidative and apoptotic testicular toxicity was evaluated by biochemical, histological and flow cytometric analyses. KEY RESULTS: Compared with controls, testes from doxorubicin-treated rats displayed impaired spermatogenesis, depleted haploid germ cell subpopulations, increased lipid peroxidation products (malondialdehyde), depressed antioxidant enzyme activities (superoxide dismutase, glutathione peroxidase and glutathione), reduced antioxidant enzyme expression (superoxide dismutase) and elevated apoptotic indexes (pro-apoptotic modulation of Bcl-2 family proteins, intensification of p53 and Apaf-1, release of mitochondrial cytochrome c, activation of caspase-3 and increase of terminal deoxynucleotidyl transferase nick-end labelling/sub-haploid cells), while EGb pretreatment effectively alleviated all of these doxorubicin-induced abnormalities in testes. CONCLUSIONS AND IMPLICATIONS: These results demonstrate that EGb protected against the oxidative and apoptotic actions of doxorubicin on testes. EGb may be a promising adjuvant therapy medicine, potentially ameliorating testicular toxicity of this anti-neoplastic agent in clinical practice.

Severe pulmonary hypertension complicates postoperative outcome of non-cardiac surgery.

BACKGROUND: Whether and how pulmonary hypertension (PH) impacts perioperative outcome in non-cardiac surgery is incompletely understood. METHODS: From November 1999, all patients undergoing non-cardiac, non-local anaesthetic surgery and ever examined by echocardiography within 30 days before surgery were screened. Those having echocardiographic pulmonary artery systolic pressure >70 mm Hg were enrolled provided they were not already intubated. Case-matched peers with normal pulmonary pressures served as controls. Perioperative outcomes were compared between the two groups, and predictors of adverse perioperative outcomes were investigated by multivariate logistic regression analysis. RESULTS: From November 1999 to August 2004, a total of 62 patients (male 38, mean age 67 yr) with PH were found. Compared with the case-matched controls, patients with PH experienced equivalently smooth operative courses, but significantly more frequent postoperative heart failure (9.7 vs 0%, P = 0.028), delayed tracheal extubation (21 vs 3%, P = 0.004), and in-hospital deaths (9.7 vs 0%, P = 0.028). Multivariate regression analysis identified emergency surgery [odds ratio (OR), 44.738; P = 0.028], coronary artery disease (CAD; OR, 9.933; P = 0.042), and systolic pulmonary artery pressure (OR, 1.101; P = 0.026) as independent predictors of postoperative mortality and surgery-specific cardiac risk level (OR, 6.791; P = 0.033) and CAD (OR 6.546, P = 0.017) as predictors of morbidity. CONCLUSION: PH is an important predictor of adverse cardiopulmonary outcome in non-cardiac surgery as reflected by markedly increased postoperative complications, especially in patients with coexistent high-risk clinical and surgical characteristics.

Cortical stabilization of beta-catenin contributes to NHERF1/EBP50 tumor suppressor function.

Anchorage-independent growth is a hallmark of tumor growth and results from enhanced proliferation and altered cell-cell and cell-matrix interactions. By using gene-deficient mouse embryonic fibroblasts (MEFs), we showed for the first time that NHERF1/EBP50 (Na/H exchanger regulator factor 1/ezrin-radixin-moesin binding phosphoprotein 50), an adapter protein with membrane localization under physiological conditions, inhibits cell motility and is required to suppress anchorage-independent growth. Both NHERF1 PDZ domains are necessary for the tumor suppressor effect. NHERF1 associates directly through the PDZ2 domain with beta-catenin and is required for beta-catenin localization at the cell-cell junctions in MEFs. Mechanistically, the absence of NHERF1 selectively decreased the interaction of beta-catenin with E-cadherin, but not with N-cadherin. The ensuing disorganization of E-cadherin-mediated adherens junctions as well as the observed moderate increase in beta-catenin transcriptional activity contributed most likely to the anchorage-independent growth of NHERF1-deficient MEFs. In vivo, NHERF1 is specifically localized at the apical brush-border membrane in intestinal epithelial cells and is required to maintain a fraction of the cortical beta-catenin at this level. Thus, NHERF1 emerges as a cofactor essential for the integrity of epithelial tissues by maintaining the proper localization and complex assembly of beta-catenin.

Liver resection for hepatocellular carcinoma in patients with cirrhosis.

BACKGROUND: Although liver resection is now a safe procedure, its role for hepatocellular carcinoma (HCC) in patients with cirrhosis remains controversial. METHODS: This study compared the results of liver resection for HCC in patients with cirrhosis over two time intervals. One hundred and sixty-one patients had resection during period 1 (1991-1996) and 265 in period 2 (1997-2002). Early and long-term results after liver resection in the two periods were compared, and clinicopathological characteristics that influenced survival were identified. RESULTS: Tumour size was smaller, indocyanine green retention rate was higher, patients were older and a greater proportion of patients were asymptomatic in period 2 than period 1. Operative blood loss, need for blood transfusion, operative mortality rate, postoperative hospital stay and total hospital costs were significantly reduced in period 2. The 5-year disease-free survival rates were 28.2 and 33.9 per cent in periods 1 and 2 respectively (P = 0.042), and 5-year overall survival rates were 45.9 and 61.2 per cent (P < 0.001). Multivariate analysis identified serum alpha-fetoprotein level, need for blood transfusion and Union Internacional Contra la Cancrum tumour node metastasis stage as independent determinants of disease-free and overall survival. CONCLUSION: The results of liver resection for HCC in patients with cirrhosis improved over time. Liver resection remains a good treatment option in selected patients with HCC arising from a cirrhotic liver.