MiR-665 Participates in the Protective Effect of Dexmedetomidine in Ischemic Stroke by ROCK2/NF-κB Axis.

Ischemic stroke is a grievous intimidation to the healthiness of sufferers. Previous studies have reported that dexmedetomidine (DEX) has a protective effect on a variety of organs. This paper aimed to explore the regulatory mechanism of DEX in ischemic stroke through miR-665/ROCK2 axis. The mice model of ischemic stroke was constructed by middle cerebral artery occlusion (MCAO). The cell model of ischemic stroke was constructed by oxygen-glucose deprivation (OGD). Cell viability and apoptosis were assessed by CCK-8 assay and flow cytometry. The expression of cytokines was detected by ELISA. Lactate dehydrogenase (LDH) concentration was evaluated by LDH kit. The cerebral infarct volume of MCAO mice was detected by TTC staining, and the apoptosis of brain cells was detected by TUNEL staining. The target relationship between ROCK2 and miR-665 was analyzed by dual-luciferase reporter assay. DEX contributed cell viability from 42 to 66% (1 µM) and restrained cell apoptosis from 26 to 18% in HT22 cells treated with OGD (P < 0.01). Meanwhile, DEX decreased the expression of cytokines and LDH concentration from 184 to 126% (P < 0.001). Moreover, the expression of miR-665 enhanced 2.9 times (P < 0.05) and the expression of ROCK2 (P < 0.05) and NF-κB p65 (P < 0.01) reduced 1.8 times and 2.2 times after DEX treatment in OGD induced HT22. And miR-665 knockdown attenuated the effect of DEX on inflammation damage (the levels of TNF-α, IL-1ß and IL-6 increased 1.36 times, 1.31 times, 1.43 time, respectively, and IL-10 decreased 1.68 times) and apoptosis from 17 to 25% (P < 0.01). MiR-665 directly targeted ROCK2 and regulated ROCK2 and NF-κB p65 expression (P < 0.01). Furthermore, ROCK2 overexpression inhibited the protective effect of DEX in HT22 induced by OGD (P < 0.001), while miR-665 overexpression reversed the regulatory of ROCK2 (P < 0.01). In vivo, DEX decreased cerebral infarction volume and inhibited apoptosis of brain cell (P < 0.001). DEX has a protective effect in ischemic stroke by promoting miR-665 expression to downregulate ROCK2/NF-κB axis, suggesting DEX has a beneficial effect on ischemic stroke and miR-665 is a conceivable target for the therapeutics and diagnosis of ischemic stroke.

[Preparation of Gold Nano-Particles as Surface-Enhanced Raman Scattering Sensors for Analysis of Banned Food Dye Chrysoidin in Yuba].

Chrysoidin is a kind of banned food dye, and it has been illegally used for coloring food. A rapid detection and quantification method is developed and applied in analysis chrysoidin in yuba. Gold nanoparticles are synthesized by using hexadecyl trimethyl ammonium bromide (CTAB) as the bifunctional ligand to link the solid substrate and the AuNPs. The laser wavelength used for quantitative is 1594 cm⁻¹. Significant differences between different concentrations of chrysoidin are verified by multiple variable analysis. A relationship between the logarithm of the concentrations and the intensity of laser is proved using univariate analysis method. The calibration curves showed good linearity in the range of 0.001-0.5 mmol · L⁻¹ with correlation coefficients r = 0.995. The method is successfully applied to the determination of chrysoidin in yuba. The average recoveries of the drugs spiked at 50 and 500 µg · g⁻¹ levels are 82.4% and 116.9%, and the relative standard deviations (RSD) are 3.8% and 4.0%. The method is simple, rapid, sensitive and accurate in the determination of chrysoidin.

Ultrasensitive electrochemical biosensor for detection of DNA from Bacillus subtilis by coupling target-induced strand displacement and nicking endonuclease signal amplification.

A simple, ultrasensitive, and specific electrochemical biosensor was designed to determine the given DNA sequence of Bacillus subtilis by coupling target-induced strand displacement and nicking endonuclease signal amplification. The target DNA (TD, the DNA sequence from the hypervarient region of 16S rDNA of Bacillus subtilis) could be detected by the differential pulse voltammetry (DPV) in a range from 0.1 fM to 20 fM with the detection limit down to 0.08 fM at the 3s(blank) level. This electrochemical biosensor exhibits high distinction ability to single-base mismatch, double-bases mismatch, and noncomplementary DNA sequence, which may be expected to detect single-base mismatch and single nucleotide polymorphisms (SNPs). Moreover, the applicability of the designed biosensor for detecting the given DNA sequence from Bacillus subtilis was investigated. The result obtained by electrochemical method is approximately consistent with that by a real-time quantitative polymerase chain reaction detecting system (QPCR) with SYBR Green.

Identification of significant genes with poor prognosis in ovarian cancer via bioinformatical analysis.

Ovarian cancer (OC) is the highest frequent malignant gynecologic tumor with very complicated pathogenesis. The purpose of the present academic work was to identify significant genes with poor outcome and their underlying mechanisms. Gene expression profiles of GSE36668, GSE14407 and GSE18520 were available from GEO database. There are 69 OC tissues and 26 normal tissues in the three profile datasets. Differentially expressed genes (DEGs) between OC tissues and normal ovarian (OV) tissues were picked out by GEO2R tool and Venn diagram software. Next, we made use of the Database for Annotation, Visualization and Integrated Discovery (DAVID) to analyze Kyoto Encyclopedia of Gene and Genome (KEGG) pathway and gene ontology (GO). Then protein-protein interaction (PPI) of these DEGs was visualized by Cytoscape with Search Tool for the Retrieval of Interacting Genes (STRING). There were total of 216 consistently expressed genes in the three datasets, including 110 up-regulated genes enriched in cell division, sister chromatid cohesion, mitotic nuclear division, regulation of cell cycle, protein localization to kinetochore, cell proliferation and Cell cycle, progesterone-mediated oocyte maturation and p53 signaling pathway, while 106 down-regulated genes enriched in palate development, blood coagulation, positive regulation of transcription from RNA polymerase II promoter, axonogenesis, receptor internalization, negative regulation of transcription from RNA polymerase II promoter and no significant signaling pathways. Of PPI network analyzed by Molecular Complex Detection (MCODE) plug-in, all 33 up-regulated genes were selected. Furthermore, for the analysis of overall survival among those genes, Kaplan-Meier analysis was implemented and 20 of 33 genes had a significantly worse prognosis. For validation in Gene Expression Profiling Interactive Analysis (GEPIA), 15 of 20 genes were discovered highly expressed in OC tissues compared to normal OV tissues. Furthermore, four genes (BUB1B, BUB1, TTK and CCNB1) were found to significantly enrich in the cell cycle pathway via re-analysis of DAVID. In conclusion, we have identified four significant up-regulated DEGs with poor prognosis in OC on the basis of integrated bioinformatical methods, which could be potential therapeutic targets for OC patients.