MiRNA-210 induces the apoptosis of neuronal cells of rats with cerebral ischemia through activating HIF-1α-VEGF pathway.

OBJECTIVE: This study aims to explore the expression of micro-ribonucleic acid (miRNA)-210 in the cerebral cortex of rat model with global cerebral ischemia, and determine its function on the regulation of the apoptosis of neuronal cells. MATERIALS AND METHODS: Rat models of global cerebral ischemia were established in vitro. Rats were euthanized at 24 h after reperfusion and the cerebral cortex was collected. The expression of miRNA-210 was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Neuronal cells were transfected by liposomes in vitro and cells were divided into neuronal cell group (group i), neuronal cell + miRNA-210 mimic group (group ii) and neuronal cell + miRNA-210 inhibitor group (group iii). The cell apoptosis rate and gene and protein expressions of HIF-1α, VEGF and Caspase-3 were measured. RESULTS: The level of miRNA-210 in rats with global cerebral ischemia was remarkably higher than that in rats from sham operation group (p<0.05). The apoptosis rate of neuronal cells was increased evidently when miRNA-210 was overexpressed, and the expressions of HIF-1α, VEGF and Caspase-3 were elevated markedly at both mRNA and protein levels. CONCLUSIONS: Our data indicate that miRNA-210 expression is upregulated in the rats with global cerebral ischemia, and the rise of miRNA-210 level increases the apoptosis of neuronal cells through the activation of HIF-1α-VEGF signaling pathway.

miR-532-5p is a prognostic marker and suppresses cells proliferation and invasion by targeting TWIST1 in epithelial ovarian cancer.

OBJECTIVE: Dysregulated miR-532-5p has been observed in epithelial ovarian cancer (EOC). However, the potential biological function and clinical significance have not been fully explained. The study aimed to investigate the prognostic value and potential role of miR-532-5p in EOC. PATIENTS AND METHODS: MiR-532-5p and Twist homolog 1 (TWIST1) mRNA expression were examined using quantitative real-time PCR. The correlation of miR­532-5p expression with clinicopathological factors was statistically analyzed. Kaplan-Meier analysis and Cox proportional hazards regression models were explored to reveal the correlations of miR-532-5p expression with survival of patients. Cell Counting Kit-8, colony formation and transwell invasion assays were performed to evaluate the effects of miR-532-5p on cell proliferation and invasion, respectively. MiR­532-5p target genes were confirmed using luciferase activity, RT-PCR and Western blot assays. RESULTS: We found that miR-532-5p was significantly decreased in EOC tissue and cell lines, and its expression levels were highly correlated with grade (p = 0.011), FIGO stage (p = 0.004) and distant metastasis (p = 0.008). In addition, overall patient survival for those with high miR-532-5p expression was significantly longer than those patients with low miR-532-5p expression (p = 0.0058). Multivariate regression analysis identified miR-532-5p down-regulation as an independent unfavorable prognostic factor in EOC patients. Function assays showed that overexpression of miR-532-5p inhibited proliferation, colony formation and invasion of EOC cells. Mechanistic investigations confirmed TWIST1 as a direct target of miR-532-5p. Further in vitro assay indicated that restored expression of TWIST1 dampened miR-532-5p-mediated suppression of tumor progression. CONCLUSIONS: Our data suggested that miR-532-5p may act not only as a novel prognostic marker, but also as a potential target for molecular therapy of EOC.

Serum miR-629 is a novel molecular marker for diagnosis and the prognosis of pancreatic cancer.

OBJECTIVE: Increasing evidence indicates that dysregulation of miRNAs is involved in tumor progression and development. We aimed to determine potential values of miR-629 as a serum diagnostic and prognostic biomarker in pancreatic cancer (PC). PATIENTS AND METHODS: MiR-629 expression levels in PC tissues and serum were measured by quantitative Real-time reverse transcription-polymerase chain reaction (qRT-PCR). Receiver operating characteristic analysis (ROC) was utilized to assess the predictive power of serum miR-629 for PC. Then, the associations of serum miR-629 expression levels with clinicopathological features and prognosis were evaluated. RESULTS: We found that the expression levels of miR-629 were significantly upregulated in both PC tissues and serum in comparison with matched normal tissues and healthy controls, respectively. Importantly, serum miR-629 could efficiently screen PC patients from healthy controls (AUC=0.765). The diagnosis capability of serum miR-629 was significantly higher than that of CA19-9, and the combination of two molecules had higher diagnosis capacity. Higher expression of serum miR-629 in PC patients was associated with advanced TNM stage (p=0.000) and distant metastasis (p=0.003). Moreover, Kaplan-Meier analysis indicated that patients with high expression of serum miR-629 had significantly shorter overall survival (p=0.0022) and disease-free survival (p=0.0003) than the low expression group. Univariate and multivariate analysis showed that serum miR-629 was a significant and independent prognostic predictor for both overall survival and disease-free survival of PC patients. CONCLUSIONS: This study suggested serum miR-629 may be a potential biomarker for the diagnosis and prognosis of PC.

Inflammatory factor in donor liver and its effect on recipient myocardial injury after liver transplantation.

OBJECTIVE: To study the inflammatory factors in donor livers and its effect on recipient myocardial injury after liver transplantation recipients. PATIENTS AND METHODS: Eighteen patients who underwent orthotopic liver transplantations between January 2014 and December 2015 in our hospital were selected. A portion of the hepatic venous blood of donor's livers was preserved in heparinized tubes after partial resection. The concentrations of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), cardiac troponin I (cTnI), creatine kinase isoenzyme (CK-MB), and the activity of lactate dehydrogenase (LDH) in serum were measured. The concentrations of TNF-α, IL-6, cTnI, and CK-MB, and the activity of LDH in serum from the central venous blood of recipients were measured at several time points. RESULTS: Persistent myocardial injuries were found in five patients, six experienced a transient increase of cardiac markers after surgery and returned to normal levels 24 h after surgery, and the others were normal. The comparison of the levels of inflammatory factors in serum between the five donors and recipients at different stages showed that the levels of myocardial markers of the donor livers which were supplied to the five cardiac injured patients were all significantly higher than those of other donor's livers, while the levels of serum inflammatory factors in recipients showed no changes during the T0-T2 stage but increased significantly during T3-T5 (p < 0.05). The cardiac function after surgery was significantly different from that before surgery and that of the recipients without myocardial injury (p < 0.05). CONCLUSIONS: Blood pressure changes before surgery may affect the levels of inflammatory factors in donor's liver and cause postoperative myocardial injury in recipients. Proper hypotensive therapy for donors before partial liver resection can prevent postoperative myocardial injury in recipients.

The research on the influences of hyperthermal perfusion chemotherapy combined with immunologic therapy on the immunologic function and levels of circulating tumor cells of the advanced colorectal cancer patients with liver metastasis.

OBJECTIVE: To investigated the influence of hyperthermal perfusion chemotherapy combined with immunologic therapy on the immunologic function and levels of circulating tumor cells of the advanced colorectal cancer patients with liver metastasis. PATIENTS AND METHODS: We enrolled 98 advanced colorectal cancer patients with liver metastasis that were admitted to this hospital for treatment and were randomly divided into two groups, the observation group (n = 49) and the control group (n = 49). We administered systemic vein chemotherapy for patients in the control group, and hyperthermal perfusion chemotherapy for the patients in the observation group in order to compare the subgroup levels of T lymphocytes, NK cells and immunoglobulin (IgG, IgA, and IgM) in the immune system of patients in both groups. We also assayed the circulating tumor cells (CTC) in the peripheral blood of patients in both groups using the cell search method, and compared the efficacy using response evaluation criteria in solid tumors and the survival rates of patients in both groups using the Kaplan-Meier method. RESULTS: After two treatment courses, the levels of CD3+, CD4+ and CD4+/CD8+ of the patients in the observation group were significantly higher than those of the control group, but the levels of CD8+ of patients in the observation group was lower than that in the control group (p< 0.05). The levels of immunoglobulins (IgG, IgA, and IgM) in the observation group were higher than the control group (p < 0.05). The levels of NK cell cells were significantly lower than the control group (p < 0.05). The objective response rate, as well as the disease control rate of the observation group, were remarkably higher than those of the control group (p < 0.05). Compared to the control group, the observation group enjoyed a prolonged survival time, higher survival rate and significantly lower positive rate of CTC (p < 0.05). CONCLUSIONS: Better efficacy and tolerance, fewer toxic and side effects, improvement in the immunologic functions of patients for the indirect anti-tumor effect, a significant decrease in CTC of patients, and a higher long-term survival rate have been achieved in the treatment with hyperthermal perfusion chemotherapy combined with immunologic therapy for the advanced colorectal cancer patients with liver metastasis. Thus, it can serve as the preferable drug for the treatment of advanced colorectal cancer with liver metastasis.

Effect of tobacco smoke on hydrogen sulfide-induced rat thoracic aorta relaxation

Levels of hydrogen sulfide (H2S), a gaseous signaling molecule, are reduced in the serum of individuals who smoke. We hypothesized that tobacco smoke influenced smooth muscle relaxation by decreasing H2S levels and this effect could also influence expression of cystathionine γ-lyase (CSE) and sulfonylurea receptor-2 (SUR-2). The aim of this study was to explore the effect of tobacco smoke on H2S-mediated rat thoracic aorta relaxation and its possible mechanism. Thirty-two Sprague-Dawley rats were divided into four groups: control (C) group, short-term smoker (SS) group, mid-term smoker (MS) group, and long-term smoker (LS) group. H2S concentrations in serum, action of H2S on rat aortic vascular relaxation, and expression of CSE and SUR-2 in thoracic aortic smooth muscle were measured. Although there was no significant difference in H2S between the C and the SS groups, concentration of H2S was significantly reduced in both the LS and MS groups compared to control (P<0.01). Furthermore, H2S was significantly lower in the LS than in the MS group (P<0.05). Rat aortic vascular relaxation was lower in all three treatment groups compared to the control, with the most significant decrease observed in the LS group (P<0.05 compared to the MS group). Expression of CSE and SUR-2 was reduced in the LS and MS groups compared to control (P<0.05), with the lowest levels observed in the LS group (P<0.05). Therefore, tobacco smoke reduced expression of CSE and SUR-2 in rat thoracic aorta, which may inhibit H2S production and vascular dilation.

The effect of EGB on proliferation of gastric carcinoma SGC7901 cells

Objective: To investigate the effect of Ginkgo biloba extract (EGB) on the proliferation and cell cycles of gastric carcinoma SGC7901 cells, and make a preliminary exploration on possible molecular mechanisms associated with its inhibitory effect. Methods: Human gastric carcinoma SGC7901 cells were cultured in vitro, and treated with various concentrations (100, 200, 300, 400 mg/L) of EGB for different incubation periods (24, 48 and 72 h). CCK-8 assay was used to detect cell proliferation and flow cytometry was performed to analyze the effect of EGB on cell cycles. In addition, mRNA and protein level of two cell cycle regulators cyclin D1 and c-myc in SGC7901 cells treated with EGB were determined using PCR and Western blot. And subcutaneous xenograft model of gastric carcinoma in nude mice was established to evaluate the anti-cancer effect of EGB in vivo. Results: The proliferation of gastric carcinoma SGC7901 cells was inhibited by EGB in dose- and time-dependent manner. Flow cytometry showed cell cycle arrest in EGB-treated cells, with increased percentage of cells in G1 phase and decreased percentage in S stage. In addition, the mRNA and protein level of cyclin D1 and c-myc genes were significantly down-regulated in cells with EGB treatment with the concentration increasing. Conclusion: EGB conferred an inhibitory effect on the proliferation of gastric carcinoma SGC7901 cells both in vitro and in vivo. The inhibitory effect was dose dependent and possibly depended on inhibiting cell cycle through G1 arrest induction by suppressing cyclin D1 and c-myc expression (AU)

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Extracellular signal-regulated protein kinase in human intractable epilepsy.

Extracellular signal-regulated kinases (ERK) such as ERK1 [p44 mitogen-activated protein kinase (MAPK)] and ERK2 (p42 MAPK) are activated in the central nervous system under physiological and pathological conditions such as ischemia and epilepsy. Our aim is to investigate ERK1, ERK2, and phosphorylated ERK (p-ERK) (Thr202/Tyr 204) expression in the temporal lobe of patients with intractable epilepsy (IE) and to explore its possible role of ERK in it. Tissue samples from temporal neocortices of 40 patients who had surgery for IE were used to detect ERK1, ERK2, and p-ERK (Thr 202/Tyr 204) expression through immunohistochemistry and western blot. We compared these tissues against 17 histological normal temporal lobes from head-trauma patients. ERK1, ERK2, and p-ERK in IE were significantly higher than those in the controls. They were mainly expressed in the cytoplasm of neurons and glial cells. There was also increased detection of p-ERK in the gliotic cortex of IE compared with the non-gliotic cortex. These findings were consistently observed in western blot and immunohistochemistry techniques. ERK expression in patients with IE was significantly increased compared with the controls. This suggested a probable role of ERK in the pathogenesis of IE.

A reovirus disease in cultured mud crab, Scylla serrata, in southern China.

A reovirus, designated mud crab reovirus (MCRV), associated with large economic losses was recently isolated from marine cultured mud crab, Scylla serrata, in southern China. The complete viral particle is 70 nm in diameter, icosahedral and non-enveloped. The virus infects connective tissue cells of the hepatopancreas, gills and intestine in mud crab and develops in the cytoplasm. Hundred per cent mortality was observed in mud crab experimentally infected by intramuscular injection, bath inoculation and oral inoculation, while cohabitation infection caused 80% mortality. The viral genome consists of 13 linear dsRNA segments, with an electrophoretic pattern 1/5/7. The results of this study suggest that the virus is highly pathogenic and can be transmitted enterically as well as via the body surface of mud crab. Although the genomic organization of this virus is different from that of the other crab reoviruses, CcRV-W2 and DpPV, all three of these reoviruses have similar electrophoresis patterns. Therefore, MCRV may be a new member of the DpPV and CcRV-W2 group.