Circ_0000615 promotes high glucose-induced human retinal pigment epithelium cell apoptosis, inflammation and oxidative stress via miR-646/YAP1 axis in diabetic retinopathy.

BACKGROUND: Diabetic retinopathy (DR), a common complication of diabetes mellitus, is the major cause of visual impairment and blindness. Circ_0000615 was found to be elevated in retina samples of diabetic patients. Hence, the detailed effects and molecular mechanisms of circ_0000615 in DN progression were explored. METHODS: The levels of circ_0000615, microRNA (miR)-646 and YAP1 (yes-associated protein 1) were detected using quantitative real-time polymerase chain reaction and Western blot assays. Cell viability, apoptosis, inflammation and reactive oxygen species (ROS) generation were determined using cell counting kit-8 assay, flow cytometry, caspase3 activity analysis, Western blot, enzyme-linked immunosorbent assay (ELISA) and Dichlorofluorescein diacetate (DCFH-DA) assay, respectively. The binding interaction between miR-646 and circ_0000615 or YAP1 was determined using dual-luciferase reporter, RNA immunoprecipitation (RIP), and RNA pull-down assays. RESULTS: Circ_0000615 was elevated in high glucose (HG)-induced human retinal pigment epithelium (HRPE) cells. Knockdown of circ_0000615 attenuated HG-triggered HRPE cell apoptosis, inflammation, and ROS generation. Mechanistically, miR-646 was confirmed to be a target of circ_0000615, inhibition of miR-646 reversed the protective effects of circ_0000615 knockdown on HG-evoked HRPE cell dysfunction. MiR-646 was verified to target YAP1, overexpression of YAP1 abolished the impairment induced by miR-646 on HG-induced HRPE cell damage. Besides that, we confirmed that circ_0000615 could regulate YAP1 expression via miR-646. CONCLUSION: Circ_0000615 contributed to HG-induced HRPE cell dysfunction via miR-646/YAP1 axis, suggesting a novel insight into the pathogenesis of DR and a potential candidate for DR treatment.

Antitumor Effects of Baicalein and Its Mechanism via TGFß Pathway in Cervical Cancer HeLa Cells.

BACKGROUND: Due to dual-regulating carcinogenesis, the TGFß pathway is an ideal and alternative tumor target. Natural flavonoids possess the similar structures to estrogen and could exert an important benefit to cervical cancer. The present study aimed to screen the inhibitor of TGFß pathway from natural flavonoids and evaluate the function and mechanism of the TGFß pathway inhibitor on cervical cancer. MATERIALS AND METHODS: The cervical cancer HeLa cells were firstly treated with different flavonoids and probed by western blot for screening the inhibitor of TGFß pathway. And then, the effect of the identified inhibitor on cell proliferation was studied by CCK-8 and clone formation assay. Then, RT-PCR and western blot assay were performed to evaluate the effect of identified inhibitor on mTOR/p70S6K pathway, and the cell migration and EMT pathway were also examined using scratching analysis and western blot assay. Finally, the role of TGFß was assessed via the classic inhibitor of TGFß/SMAD pathway. RESULTS: Screening data by western blot assay showed that baicalein displayed the best inhibitor effect on TGFß expression. CCK-8 and clone formation assay showed that baicalein inhibited the cell proliferation and clone cell number. RT-PCR and western bolt for probing mTOR, p70S6K, and 4EBP1 revealed that baicalein could suppress their expression and phosphorylation. The scratching analysis and western blot assay displayed that baicalein inhibited the cell migration and EMT progression in HeLa. The use of SB431542, a TGFß inhibitor, revealed that TGFß was crucial to baicalein-regulating cell proliferation and migration in HeLa cells. CONCLUSION: Baicalein, a medicine agent screened from natural flavonoids targeting TGFß pathway, could suppress mTOR/p70S6K pathway-mediated cell proliferation and EMT pathway-related migration via TGFß pathway in cervical cancer HeLa cells.

Catalytic pyrolysis and liquefaction behavior of microalgae for bio-oil production.

Microalgae bio-oil production is related to the sustainable use of world energy in the future. In the present work, catalytic pyrolysis and liquefaction behavior of microalgae for bio-oil production were investigated. The results show that the rare earth compounds as catalysts contributed to significantly accelerating the pyrolysis of microalgae via reducing the activation energy of pyrolysis process. Ce(II)/HZSM-5 presented the optimal catalytic pyrolysis and liquefaction effects by helping cut the microalgae molecule chains. The maximum bio-oil yield amounted to 49.71 wt% at the catalyst concentration of 5 wt%. The chemical components of the Spirulina bio-oil were composed of carboxylic acids, ketones, olefins, amides, ethers, esters, and partially cyclic N-containing compounds. Although the combustion performances of the Spirulina bio-oil are worse than those of the diesel fuel, it is superior to the reported rice husk bio-oil, suggesting a promising potential application prospect.

Polysaccharides from Rhizoma Panacis Majoris and its anti-oxidant activity.

Response surface method (RSM) was employed to optimize the extraction conditions of polysaccharides from Rhizoma Panacis Majoris (the rhizomes of Panax japonicus C. A. Mey. var. major (Burk.) C. Y. Wu et K. M. Feng) (RPMP), a well-known Chinese traditional medicine. In order to obtain the optimal processing parameters, a three-variable Box-Behnken designs (BBD) were applied for experimental designs. RSM analysis indicated the good correspondence between experimental and predicted values, the optimal conditions for the yield of polysaccharides were as follows: the ultrasound time is 31.15 min, extraction temperature is 92.50 °C, and the ratio of water to raw material is 40 mL/g. The maximum value (13.87 ± 0.16%) of the yield of polysaccharides was obtained under these optimal conditions. The molecular weight (MW) was determined to be 1.48 × 10(5)(±0.39%)Da by HPSEC-MALLS-RID chromatography system. FT-IR spectra demonstrated obvious characteristic peaks of polysaccharides. The antioxidant activities of RPMP were investigated including scavenging activity of hydrogen radicals, ABTS radicals, and free radicals of superoxide anion in vitro, and the results exhibited that RPMP had a good potential for antioxidant.