miR-302a-3p suppresses melanoma cell progression via targeting METTL3.

The miRNA-302 family plays a critical role in carcinogenesis. As an enzyme that regulates the N6-methyladenosine modification, methyltransferase-like 3 (METTL3) plays important roles in the development and progression of various tumours. However, the upstream regulatory mechanisms of METTL3 in melanoma have not yet been fully investigated. Herein, we investigated the functions of miR-302a-3p and its target RNA METTL3 on proliferation, apoptosis, and invasion of melanoma. Quantitative real-time PCR and immunofluorescence staining were used to measure the expression of METTL3 mRNA and protein level after transfection. miR-302a-3p expression was determined by quantitative reverse transcription-PCR. The cell proliferation, cell cycle progression, apoptosis, colony formation, migration, and cell invasion ability were determined using MTT assay, propidium iodide (PI) staining, Annexin V/PI flow cytometry, plate clone assay, and Transwell migration and invasion assays, respectively. Melanoma cell metastasis was also evaluated using an in vivo model. The effect of METTL3 on the phosphorylation of PI3K and AKT was measured with western blot analysis. Our results showed that miR-302a-3p was significantly downregulated in melanoma and exerted a tumour suppressive role against melanoma progression. We identified METTL3 as a direct target of miR-302a-3p in melanoma cells using bioinformatics analysis and luciferase assay. Furthermore, the enforced overexpression of METTL3 promoted the proliferation, cell cycle progression, cell invasion, migration, expression of epithelial-to-mesenchymal transition markers, and the PI3K-AKT signalling pathway as well as suppressed the apoptosis of melanoma cells. Meanwhile, silencing the expression of METTL3 with specific shRNA demonstrated reverse outcomes of the above phenotypes in melanoma cells. By rescue experiments, we found that the restoration of METTL3 expression in miR-302a-3p-overexpressing melanoma cells successfully recovered the miR-302a-3p-mediated melanoma suppression. The in vivo results also showed that miR-302a-3p substantially inhibited melanoma cell growth and metastasis. In summary, this study demonstrated that miR-302a-3p targets METTL3 and plays tumour suppressive roles in the proliferation, apoptosis, invasion, and migration of melanoma cells.

Melatonin ameliorates microvessel abnormalities in the cerebral cortex and hippocampus in a rat model of Alzheimer's disease.

Melatonin can attenuate cardiac microvascular ischemia/reperfusion injury, but it remains unclear whether melatonin can also ameliorate cerebral microvascular abnormalities. Rat models of Alzheimer's disease were established by six intracerebroventricular injections of amyloid-beta 1-42, administered once every other day. Melatonin (30 mg/kg) was intraperitoneally administered for 13 successive days, with the first dose given 24 hours prior to the first administration of amyloid-beta 1-42. Melatonin ameliorated learning and memory impairments in the Morris water maze test, improved the morphology of microvessels in the cerebral cortex and hippocampus, increased microvessel density, alleviated pathological injuries of cerebral neurons, and decreased the expression of vascular endothelial growth factor and vascular endothelial growth factor receptors 1 and 2. These findings suggest that melatonin can improve microvessel abnormalities in the cerebral cortex and hippocampus by lowering the expression of vascular endothelial growth factor and its receptors, thereby improving the cognitive function of patients with Alzheimer's disease. This study was approved by the Animal Care and Use Committee of Jinzhou Medical University, China (approval No. 2019015) on December 6, 2018.

[Angiotensin II type 1 receptor autoantibody induces myocardial fibrosis by activating cardiac fibroblasts].

Myocardial fibrosis (MF) is an important pathological process of cardiac remodeling in patients with heart failure; however its etiology has not been clear. It has been known that the angiotensin II type 1 receptor autoantibody (AT1-AA) is present in patients with heart failure, but it is unclear whether this antibody directly causes MF. In this study, we investigated the role of AT1-AA in MF and its effects on cardiac fibroblasts (CFs). The AT1-AA positive rat model was established by active immunization method, and the measurement of indexes were made in the 8th week after active immunity. The results of heart echocardiography showed that the cardiac systolic and diastolic functions of AT1-AA positive rats were impaired with reduced left ventricular wall thickness and enlarged heart chambers. HE staining results showed that the myocardial fibers were disorganized and ruptured, and Masson staining revealed that the area of collagen fibers around the myocardium and coronary arteries was significantly increased in AT1-AA positive group compared with that of the control group (P < 0.05). Moreover, primary CFs isolated from neonatal rats were cultured and treated with AT1-AA for 48 h. CCK-8 and immunofluorescence staining results showed that AT1-AA enhanced proliferation rate of CFs (P < 0.001), and Western blot results showed that AT1-AA significantly increased expressions of collagen I (Col I), Col III, matrix metalloproteinase-2 (MMP-2) and MMP-9 in CFs (all P < 0.05). Taken together, these results suggest that AT1-AA may induce MF and cardiac dysfunction via activating CFs.

Stability of ß-carotene microcapsules with Maillard reaction products derived from whey protein isolate and galactose as coating materials.

The stability of ß-carotene microcapsules using Maillard reaction products (MRPs) derived from whey protein isolate (WPI) and galactose as coating materials, was studied under the varying environmental conditions of temperature, pH, air, incandescent light, and ultraviolet (UV) light. Scanning electron microscopy showed that microcapsules prepared by WPI-galactose MRPs displayed a smooth and less concave-convex surface and that the particle size (D50) of the microcapsules made with WPI-galactose MRPs was smaller than those made with WPI-galactose mixture. The storage stability of ß-carotene microencapsulated in WPI-galactose MRPs was remarkably better than that of ß-carotene microencapsulated in the WPI-galactose mixture and that of ß-carotene crystal, in respect of temperature, pH, air, incandescent light, and UV light measurements. When the storage temperature was increased from 5 to 105 °C, the retention rate of ß-carotene microcapsules significantly decreased (P<0.05). When pH values were increased from 1 to 12, the ß-carotene retention rate of the microcapsules significantly increased and afterward decreased. Compared with the retention rate of ß-carotene microencapsulated in a WPI-galactose mixture, the retention rate of ß-carotene microencapsulated in WPI-galactose MRPs was at a maximum between pH 8 and 9. Under the actions of air, incandescent light, and UV light, the retention rates of ß-carotene microcapsules in WPI-galactose MRPs and WPI-galactose mixture, as well as in ß-carotene crystal, decreased significantly as the storage time increased (P<0.05). Therefore, the use of WPI-galactose MRPs as coating materials can aid in improving the storage stability of ß-carotene microcapsules.

Development of a test strip for rapid detection of lactoperoxidase in raw milk.

Traditional methods for detecting lactoperoxidase (LP) are complex and time-consuming, so a test strip was made based on the enzymatic reaction principle to enable quick and convenient detection of LP in raw milk. In this study 0.1 mol/L citric acid (CA)/0.2 mol/L disodium hydrogen phosphate (NaP) buffer solution (pH 5.0), 22 mmol/L 3,3',5,5'-tetramethylbenzidine (TMB), 0.6 mmol/L hydrogen peroxide (H2O2), and 0.5% Tween-20 or 0.3% cetyltrimethyl ammonium bromide (CTAB) were optimal for preparing a quick, sensitive, and accurate LP test strip. The coefficient of variation (CV) of the estimated LP concentrations ranged from 2.47% to 6.72% and the minimum LP concentration detected by the test strip was 1-2 mg/L. Estimates of active LP in sixteen raw milk samples obtained using the test strip or the TMB method showed a good correlation (r=0.9776). So the test strip provides a quick, convenient, and accurate method for detecting the LP concentration of raw milk.

[Estimation of sparse vegetation coverage in arid region based on hyperspectral mixed pixel decomposition].

Based on Hyperion hyperspectral image data, the image-derived shifting sand, false-Gobi spectra, and field-measured sparse vegetation spectra were taken as endmembers, and the sparse vegetation coverage (< 40%) in Minqin oasis-desert transitional zone of Gansu Province was estimated by using fully constrained linear spectral mixture model (LSMM) and non-constrained LSMM, respectively. The results showed that the sparse vegetation fraction based on fully constrained LSMM described the actual sparse vegetation distribution. The differences between sparse vegetation fraction and field-measured vegetation coverage were less than 5% for all samples, and the RMSE was 3.0681. However, the sparse vegetation fraction based on non-constrained LSMM was lower than the field-measured vegetation coverage obviously, and the correlation between them was poor, with a low R2 of 0.5855. Compared with McGwire's corresponding research, the sparse vegetation coverage estimation in this study was more accurate and reliable, having expansive prospect for application in the future.