miRNA identification by nuclease digestion in ELISA for diagnosis of osteosarcoma.

Osteosarcoma is a bone cancer formed by the cells of the bone. Children, young adults, and teens are highly affected by osteosarcoma. Early identification of osteosarcoma is mandatory to improve the treatment and increase the lifespan of the patients. MicroRNA-195 (miR-195) was shown to be a suitable biomarker for osteosarcoma, and the present study describes a sensitive method of miR-195 identification by nuclease digestion in ELISA to detect and quantify the level of miR-195. S1 nuclease catalyzed endo- and exonucleolytic digestion of single-stranded (ss) RNA and DNA on ELISA polystyrene substrate, which helped to identify duplexed miR-195. This method selectively and specifically identified miR-195 without any biofouling interactions and reached the limit of detection at 10 fM within the range from 10 fM to 10 nM. Due to complete digestion of ssDNA, single- and triple-mismatched sequences failed to increase the ELISA signal, indicating specific miRNA detection. Furthermore, human serum spiked with miR-195 did not interfere with the detection, confirming selective identification. This method identified miR-195 at a lower level and will help to diagnose earlier stages of osteosarcoma.

Azilsartan prevented AGE-induced inflammatory response and degradation of aggrecan in human chondrocytes through inhibition of Sox4.

Advanced glycation end products (AGEs)-induced inflammation and degradation of aggrecan in human chondrocytes play an important role in the progression and development of osteoarthritis (OA). Azilsartan, an angiotensin II receptor antagonist, has been licensed for the treatment of high blood pressure. However, the effects of Azilsartan in OA and AGEs-induced damages in chondrocytes have not been previously reported. The injured chondrocytes model was established by incubating with 5 µmol/L AGEs. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide was used to evaluate the cell viability of treated SW1353 cells. The gene expression levels of interleukin-1α (IL-1α), tumor necrosis factor-ß (TNF-ß), IL-6, a disintegrin-like and metallopeptidase with thrombospondin type motif-4 (ADAMTS-4), ADAMTS-5, Aggrecan, and Sox-4 were evaluated using quantitative real-time polymerase chain reaction and their protein levels were determined using enzyme-linked immunosorbent assay or Western blot analysis. Mitogen-activated protein kinase p38 pathway was surveyed using phosp-p38 level and its specific inhibitor SB203580 was employed to block the p38 pathway. The overexpression of Sox4 plasmid was transfected into SW1353 cells to assess its regulation on ADAMTS-4 and ADAMTS-5. Azilsartan reduced AGEs-induced production of proinflammatory cytokines, such as IL-1α, TNF-ß, and IL-6. Azilsartan prevented AGEs-induced expressions of ADAMTS-4 and ADAMTS-5 as well as the reduction of aggrecan. Mechanistically, AGEs treatment increased the expression of Sox4 in a dose-dependent manner. AGE treatment increased the level of phosphorylated p38. However, treatment with the p38 inhibitor SB203580 inhibited AGEs-induced expression of Sox4, suggesting that AGEs-induced expression of Sox4 is mediated by p38. Furthermore, Azilsartan suppressed AGEs-induced phosphorylation of p38 and expression of Sox4. Finally, the overexpression of Sox4 abolished the inhibitory effects of Azilsartan against the expressions of ADAMTS-4 and ADAMTS-5. Azilsartan treatment prevented AGEs-induced inflammatory response and degradation of aggrecan through inhibition of Sox4.

Web-Based Technology for Remote Viewing of Radiological Images: App Validation.

BACKGROUND: Internet technologies can create advanced and rich web-based apps that allow radiologists to easily access teleradiology systems and remotely view medical images. However, each technology has its own drawbacks. It is difficult to balance the advantages and disadvantages of these internet technologies and identify an optimal solution for the development of medical imaging apps. OBJECTIVE: This study aimed to compare different internet platform technologies for remotely viewing radiological images and analyze their advantages and disadvantages. METHODS: Oracle Java, Adobe Flash, and HTML5 were each used to develop a comprehensive web-based medical imaging app that connected to a medical image server and provided several required functions for radiological interpretation (eg, navigation, magnification, windowing, and fly-through). Java-, Flash-, and HTML5-based medical imaging apps were tested on different operating systems over a local area network and a wide area network. Three computed tomography colonography data sets and 2 ordinary personal computers were used in the experiment. RESULTS: The experimental results demonstrated that Java-, Flash-, and HTML5-based apps had the ability to provide real-time 2D functions. However, for 3D, performances differed between the 3 apps. The Java-based app had the highest frame rate of volume rendering. However, it required the longest time for surface rendering and failed to run surface rendering in macOS. The HTML5-based app had the fastest surface rendering and the highest speed for fly-through without platform dependence. Volume rendering, surface rendering, and fly-through performances of the Flash-based app were significantly worse than those of the other 2 apps. CONCLUSIONS: Oracle Java, Adobe Flash, and HTML5 have individual strengths in the development of remote access medical imaging apps. However, HTML5 is a promising technology for remote viewing of radiological images and can provide excellent performance without requiring any plug-ins.