MicroRNA-193a-5p exerts a tumor suppressor role in glioblastoma via modulating NOVA1.

BACKGROUND AND OBJECTIVES: Glioblastoma (GBM) is the most common and lethal of intracranial tumors, which is characterized by extensive proliferation and the diffused invasion of tumor cells. MicroRNA-193a-5p (miR-193a-5p) have been demonstrated previously as a functional suppressor in the development and progression of various cancers. The current study aimed to investigate whether miR-193a-5p influences cell proliferation and migration through the mitogen-activated protein kinase/extracellular signal-regulated kinase signaling pathway by targeting neuro-oncological ventral antigen 1 (NOVA1) in glioblastoma. MATERIALS AND METHODS: The miR-193a-5p expression was detected by quantitative real-time polymerase chain reaction assay in GBM tissues and cell lines. Cell Counting Kit-8 assay, colony formation analysis, wound-healing, and transwell invasion assays were performed to evaluate cell proliferation, colony formation, migration, and invasion, respectively. Western blot analysis and luciferase reporter gene assay were performed to verify the downstream target gene of miR-193a-5p. RESULTS: The expression of miR-193a-5p was significantly downregulated in GBM tissues and cell lines. Kaplan-Meier analysis showed that patients with low miR-193a-5p expression had a shorter disease-free survival (P < 0.05). Functionally, miR-193a-5p overexpression dramatically suppressed the proliferation, colony formation, migration, and invasion in glioma cells. Bioinformatics prediction and a luciferase assay confirmed that NOVA1 was a direct functional target of miR-193a-5p. Moreover, ectopic expression of NOVA1 could partially reverse the inhibitory effects of miR-193a-5p on glioma cell proliferation, colony formation, migration, and invasion. NOVA1 overexpression abrogated the inhibitory effect of miR-193a-5p on the PTEN/PI3k/AKT pathway. CONCLUSION: Taken together, our findings suggested that miR-193a-5p functions as a tumor suppressor in glioma cells by directly targeting NOVA1.

Clipping versus Coiling for Ruptured Intracranial Aneurysms: A Meta-Analysis of Randomized Controlled Trials.

BACKGROUND: The treatment strategies of ruptured intracranial aneurysms (RIAs) include surgical clipping and endovascular coiling, and the efficacy and safety of clipping versus coiling are yet controversial. OBJECTIVE: To summarize the available randomized controlled trials to determine the optimal treatment method for patients with RIA. METHODS: PubMed, Embase, and the Cochrane Library were systematically searched for randomized controlled trials published up to September 5, 2017. The summary analysis was performed using a random-effects model. The primary outcomes included poor outcome, mortality, and rebleeding, whereas the secondary outcomes included complete occlusion, incomplete occlusion, severe disability, and vegetative state. RESULTS: We identified 5 trials with data collected from 2883 patients. The summary results indicated that surgical clipping in patients was associated with a high incidence of poor outcome (relative risk [RR], 1.34; 95% confidence interval [CI], 1.18-1.51; P < 0.001), whereas no significant effect was observed on mortality (RR, 1.09; 95% CI, 0.79-1.49; P = 0.608) and rebleeding (RR, 0.65; 95% CI, 0.20-2.06; P = 0.460) compared with endovascular coiling. Furthermore, we noted that surgical clipping significantly increased the incidence of complete occlusion compared with endovascular coiling (RR, 1.30; 95% CI, 1.09 1.55; P = 0.004). Conversely, surgical clipping was associated with a low incidence of incomplete occlusion (RR, 0.67; 95% CI, 0.45-0.99; P = 0.044). No significant differences were noted between surgical clipping and endovascular coiling with respect to the outcomes of severe disability (RR, 1.39; 95% CI, 0.90-2.16; P = 0.140) and vegetative state (RR, 1.35; 95% CI, 0.84-2.17; P = 0.213). CONCLUSIONS: This meta-analysis provides moderate evidence that surgical clipping has few benefits than endovascular coiling for the treatment of RIA.

Carnosic Acid Mitigates Early Brain Injury After Subarachnoid Hemorrhage: Possible Involvement of the SIRT1/p66shc Signaling Pathway.

Carnosic acid (CA) has been reported to exhibit a variety of bioactivities including antioxidation, neuroprotection, and anti-inflammation; however, the impact of CA on subarachnoid hemorrhage (SAH) has never been elucidated. The current study was undertaken to explore the role of CA in early brain injury (EBI) secondary to SAH and the underlying mechanisms. Adult male Sprague-Dawley rats were perforated to mimic a clinical aneurysm with SAH. CA or vehicle was administered intravenously immediately after the SAH occurred. Mortality, SAH grade, neurologic function scores, brain water content, Evans blue extravasation, and the levels of reactive oxygen species (ROS) levels in the ipsilateral cortex were determined 24 h after the SAH occurred. Western blot, immunofluorescence, Fluoro-Jade C (FJC) and TUNEL staining were also performed. Our results showed that CA decreased ROS levels, alleviated brain edema and blood-brain barrier permeability, reduced neuronal cell death, and promoted neurologic function improvement. To probe into the potential mechanisms. We showed that CA increased SIRT1, MnSOD, and Bcl-2 expression, as well as decreased p66shc, Bax, and cleaved caspase-3 expression. Interestingly, sirtinol, a selective inhibitor of SIRT1, abolished the anti-apoptotic effects of CA. Taken together, these data revealed that CA has a neuroprotective role in EBI secondary to SAH. The potential mechanism may involve suppression of neuronal apoptosis through the SIRT1/p66shc signaling pathway. CA may provide a promising therapeutic regimen for management of SAH.

LncRNA CDKN2BAS predicts poor prognosis in patients with hepatocellular carcinoma and promotes metastasis via the miR-153-5p/ARHGAP18 signaling axis.

BACKGROUND: Growing evidence shows that long noncoding RNAs (lncRNAs) play a crucial role in cancer progression. However, whether lncRNA CDKN2BAS is involved in human hepatocellular carcinoma (HCC) metastasis remains unclear. METHODS: Human lncRNA microarray analysis was performed to detect differential expression levels of lncRNAs in metastatic HCC tissues. Effects of CDKN2BAS on cell proliferation, migration, and apoptosis were determined by MTT assay, colony formation assay, migration assay, scratch assay, and flow cytometry. The xenograft experiment was used to confirm the effect of CDKN2BAS on HCC in vivo. qRT-PCR and Western blot were performed to determine the expression levels of mRNAs and proteins. Luciferase reporter assay was used to identify the specific target relationships. RESULTS: CDKN2BAS was remarkably up-regulated in metastatic HCC tissues compared with the adjacent non-tumor tissues. CDKN2BAS promotes HCC cell growth and migration in vitro and in vivo. Additionally, CDKN2BAS upregulated the expression of Rho GTPase activating protein 18 (ARHGAP18) by sponging microRNA-153-5p (miR-153-5p), and thus promoted HCC cell migration. Besides, CDKN2BAS downregulated the expression of Krüppel-like factor 13 (KLF13) and activated MEK-ERK1/2 signaling, thus reducing apoptosis in HCC cells. CONCLUSIONS: Our study revealed that lncRNA CDKN2BAS promotes HCC metastasis by regulating the miR-153-5p/ARHGAP18 signaling.

Inhibition of microRNA-299-5p sensitizes glioblastoma cells to temozolomide via the MAPK/ERK signaling pathway.

Glioblastomas (GBMs) are a lethal class of brain cancer, with a median survival <15 months in spite of therapeutic advances. The poor prognosis of GBM is largely attributed to acquired chemotherapy resistance, and new strategies are urgently needed to target resistant glioma cells. Here we report a role for miR-299-5p in GBM. The level of miR-299-5p expression was detected in glioma specimens and cell lines by qRT-PCR. Luciferase reporter assays and Western blots were performed to verify GOLPH3 as a direct target of miR-299-5p. In vitro cell proliferation, invasion, cell cycle distribution, and apoptosis were assessed to determine whether or not miR-299-5p knockdown sensitized GBM cells to temozolomide (TMZ). We demonstrated that miR-299-5p levels were up-regulated in the GBM groups compared with the normal control group. The highest expression of miR-129-5p occurred in the highest GBM stage. miR-299-5p knockdown significantly inhibited the MAPK/extracellular signal-regulated kinase (ERK) signaling pathway. We also showed that miR-299-5p knockdown enhanced sensitivity of GBM cells to TMZ both in vitro and in vivo by inhibiting cell proliferation and invasion and promoting apoptosis. In addition, we demonstrated that GOLPH3 is a novel functional target of miR-299-5p GOLPH3 regulates the MAPK/ERK axis under miR-299-5p regulation. In conclusion, we identified a link between miR-299-5p expression and the GOLPH3/MAPK/ERK axis, thus illustrating a novel role for miR-299-5p in GBM.

Ideal Cardiovascular Health Metrics Associated with Reductions in the Risk of Extracranial Carotid Artery Stenosis: a Population-based Cohort Study.

The cardiovascular health (CVH) metrics are closely related to the risk of stroke. Extracranial carotid artery stenosis (ECAS) represents an important risk factor for ischemic stroke. The present study aims to explore the longitudinal effect of the baseline CVH metrics on the development of ECAS. Totally 5,440 participants were randomly enrolled in the Asymptomatic Polyvascular Abnormalities Community study from 2010 to 2011. Information regarding the seven CVH metrics was collected at baseline. ECAS was assessed by performing carotid duplex sonography at baseline (2010-2011) and during the follow-up (2012-2013). Finally 3,487 subjects were included, and 976 participants developed ECAS during the 2-year follow-up. The optimum CVH status was associated with a 42% (95% confidence interval: 0.40-0.85) decreased risk of the incidence of ECAS after adjusting for age, sex, weight, education, income, alcohol use, waist-hip ratio, triglycerides, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, uric acid, homocysteine, and C-reactive protein. Ideal physical activity, total cholesterol and fasting blood glucose were independent protective factors of ECAS. In this cohort study, the ideal baseline CVH status was negatively associated with the occurrence of ECAS during the follow-up. This study provides practical insight for further developing effective screening strategies or implementing the best medical treatment.

Circular RNA hsa_circ_0021001 in peripheral blood: a potential novel biomarker in the screening of intracranial aneurysm.

Circular RNAs (circRNAs) in the peripheral blood have been reported to be associated with cancer. However, there are few studies about circRNAs in intracranial aneurysms (IA). The purpose of the current study was to investigate the characteristic expression of circular RNA hsa_circ_0021001 in the peripheral blood of patients with intracranial aneurysms and its potential as a diagnostic biomarker for IA. In this study, a cohort of 223 cases of IA patients who were admitted in the department of neurosurgery in the First People's Hospital of Wenling from January 2009 to July 2012 were collected as the experimental group, and 131 healthy volunteers over the same period served as the control group. Peripheral blood of each subject in both groups was collected on an empty stomach. The expression of hsa_circ_0021001 in peripheral blood was detected by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and the difference was analyzed by paired t-test. The effectiveness of hsa_circ_0021001 in the diagnosis of IA was assessed by ROC curve. Multivariate Cox proportional hazards regression analysis was used to analyze the prognosis. Hsa_circ_0021001 level in the peripheral blood of IA patients was relatively lower than that in the control group (P=0.002). The area under ROC (AUC) was 0.87, indicating that hsa_circ_0021001 was highly effective in the diagnosis of IA. In addition, hsa_circ_0021001 expression was correlated with aneurysm rupture, Hunt, Hess level, and timing of surgery (P= 0.041, 0.013, and 0.001, respectively). Patients with high expression of hsa_circ_0021001 had longer disease-free survival (DFS) and overall survival (OS) (P < 0.05). We found for the first time that hsa_circ_0021001 decreased significantly in the peripheral blood of IA patients, which suggested that hsa_circ_0021001 might be used as a potential novel marker for the diagnosis of IA.

Prevalence of herpes and respiratory viruses in induced sputum among hospitalized children with non typical bacterial community-acquired pneumonia.

OBJECTIVE: Few comprehensive studies have searched for viruses in infants and young children with community-acquired pneumonia (CAP) in China. The aim of this study was to investigate the roles of human herpes viruses (HHVs) and other respiratory viruses in CAP not caused by typical bacterial infection and to determine their prevalence and clinical significance. METHODS: Induced sputum (IS) samples were collected from 354 hospitalised patients (infants, n = 205; children, n = 149) with respiratory illness (CAP or non-CAP) admitted to Wenling Hospital of China. We tested for HHVs and respiratory viruses using PCR-based assays. The epidemiological profiles were also analysed. RESULTS: High rate of virus detection (more than 98%) and co-infection (more than 80%) were found among IS samples from 354 hospitalised infants and children with respiratory illness in this study. Of 273 CAP samples tested, CMV (91.6%), HHV-6 (50.9%), RSV (37.4%), EBV (35.5%), HBoV (28.2%), HHV-7 (18.3%) and rhinovirus (17.2%) were the most commonly detected viruses. Of 81 non- CAP samples tested, CMV (63%), RSV (49.4%), HHV-6 (42%), EBV (24.7%), HHV-7 (13.6%) and HBoV (8.6%) were the dominant viruses detected. The prevalence of several viral agents (rhinovirus, bocavirus, adenovirus and CMV) among IS samples of CAP were significantly higher than that of non-CAP control group. We also found the prevalence of RSV coinfection with HHVs was also higher among CAP group than that of non-CAP control. CONCLUSIONS: With sensitive molecular detection techniques and IS samples, high rates of viral identification were achieved in infants and young children with respiratory illness in a rural area of China. The clinical significance of rhinovirus, bocavirus, adenovirus and HHV (especially CMV) infections should receive greater attention in future treatment and prevention studies of CAP in infants and children.

[Expression of recombinant VP2 gene in insect sf9 cells and screening of clinical specimens].

OBJECTIVE: To clone and express VP, gene from HBoV, and the expressed VP, protein was as the antigen in order to detect serum from children in Wenling area with lower respiratory tract infections. METHODS: The VP, gene was recombined with the genome of Baculovirus, which infected the insect cell. The fusion protein with HA tag was applied to confirm the specificity of expressed protein. Furthermore, the recombinant protein was observed using electron microscopy. The 176 serum from children in Wenling area with lower respiratory tract infections was screened using Western blot. RESULTS: The expressed VP2 protein was more than 60% in total proteins from insect cell, and MWt about 60 x 10(3). The virus-like particle (VLP) was observed using electron microscopy, and size about 20 nm. The 176 serum from children in wenling area with lower respiratory tract infections was screened using Western blot. The HBoV positive rate was 2.28% (4/176). CONCLUSION: The VP2 protein from human bocavirus was expressed in insect cell successfully. Through HA tag the VP2 protein was specific, and then the assay using SDS-PAGE with Western blot could detect and screen the antibody in serum from children with lower respiratory tract infections rapidly and accurately.

[Isolation and cell culture of human bocavirus (HBoV) by human bronchial epithelial cell lines].

OBJECTIVE: To investigate pave a way for studying pathogenicty of HBoV. METHODS: Isolation and cell culture of HBoV by human bronchial epithelial cell line, which was founded in our laboratory. The morphology of the virus were primarily studied with a transmission electron microscope. In addition, transcript mRNA was detected in human bronchial epithelial cells, which was passaged and infected within HBoV, using the reverse-transcription polymerase chain reaction (RT-PCR). The amplified products nucleotide sequence of HBoV were sequencing and sequence analysis. RESULTS: Cytopathic effect (CPE) was observed after the aseptic residue of filtration of 2 case sputum specimens with HBoV, which was inoculated to the human bronchial epithelial cell line. The virus particles were observed in the cytoplasm, which were hexagonal or spherical in shape and 18-26 nm in diameter,bulk was 20 nm. cDNA amplicon obtained 295 bp fragment results of electrophoresis bands as same as NS1 region of the conserved matrix gene of publish sequence of HboV. PCR products nucleotide sequence of HboV were compared with corresponding HboV GeneBank sequences. The comparison/alignment and construction of phylogenetic trees also point to an affiliation of the parvovirus to the species HBoV. CONCLUSION: Isolation and identification of HBoV could be done in the human bronchial epithelial cell, and we found some characterizing CPE in the human bronchial epithelial cell after HBoV infection. The above studies pave a way for studying pathogenicty of human bocavirus.