Apoptosis-promoting properties of miR-3074-5p in MC3T3-E1 cells under iron overload conditions.

BACKGROUND: Iron overload can promote the development of osteoporosis by inducing apoptosis in osteoblasts. However, the mechanism by which miRNAs regulate apoptosis in osteoblasts under iron overload has not been elucidated. METHOD: The miRNA expression profile in MC3T3-E1 cells under iron overload was detected by next generation sequencing. qRT-PCR was used to determine the expression of miR-3074-5p in MC3T3-E1 cells under iron overload. The proliferation of MC3T3-E1 cells was tested using CCK-8 assays, and apoptosis was measured using flow cytometry. The miRanda and TargetScan databases were used to predict the target genes of miR-3074-5p. Interaction between miR-3074-5p and the potential target gene was validated by qRT-PCR, luciferase reporter assay and western blotting. RESULTS: We found that iron overload decreased the cell viability and induced apoptosis of MC3T3-E1 cells. The results of next generation sequencing analysis showed that miR-3074-5p expression was significantly increased in MC3T3-E1 cells under iron overload conditions, which was confirmed by further experiments. The inhibition of miR-3074-5p attenuated the apoptosis of iron-overloaded MC3T3-E1 cells. Furthermore, the expression of Smad4 was decreased and was inversely correlated with miR-3074-5p expression, and overexpression of Smad4 partially reversed the viability inhibition of iron-overloaded MC3T3-E1 cells by relieving the suppression of ERK, AKT, and Stat3 phosphorylation, suggesting its regulatory role in the viability inhibition of iron-overloaded MC3T3-E1 cells. The luciferase reporter assay results showed that Smad4 was the target gene of miR-3074-5p. CONCLUSION: miR-3074-5p functions as an apoptosis promoter in iron-overloaded MC3T3-E1 cells by directly targeting Smad4.

Clock gene Bmal1 controls diurnal rhythms in expression and activity of intestinal carboxylesterase 1.

OBJECTIVES: We aimed to characterize diurnal rhythms in CES1 expression and activity in mouse intestine, and to investigate a potential role of the core clock gene Bmal1 in generating diurnal rhythms. METHODS: The regulatory effects of intestinal Bmal1 on diurnal CES1 expression were assessed using intestine-specific Bmal1 knockout (Bmal1iKO) mice and colon cancer cells. The relative mRNA and protein levels were determined by qPCR and Western blotting, respectively. Metabolic activity of CES1 in vitro and in vivo were determined by microsomal assays and pharmacokinetic studies, respectively. Transcriptional gene regulation was investigated using luciferase reporter assay. KEY FINDINGS: Total CES1 protein varied significantly according to time of the day in wild-type (Bmal1fl/fl) mice, peaking at ZT6. Of detectable Ces1 genes, Ces1d mRNA displayed a robust diurnal rhythm with a peak level at ZT6, whereas mRNAs of Ces1e, 1f and 1g showed no rhythms in wild-type mice. Loss of intestinal Bmal1 reduced the levels of total CES1 protein and Ces1d mRNA, and blunted their diurnal rhythms in mice. In vitro microsomal assays indicated that intestinal metabolism of mycophenolate mofetil (MMF, a known CES1 substrate) was more extensive at ZT6 than at ZT18. ZT6 dosing of MMF to wild-type mice generated a higher systemic exposure of mycophenolic acid (the active metabolite of MMF) as compared with ZT18 dosing. Intestinal ablation of Bmal1 down-regulated CES1 metabolism at ZT6, and abolished its time-dependency both in vitro and in vivo. Furthermore, Ces1d/CES1 rhythmicity and positive regulation of Ces1d/CES1 by BMAL1 were confirmed in CT26 and Caco-2 cells. Mechanistically, BMAL1 trans-activated Ces1d/CES1 probably via binding to the E-box elements in the gene promoters. CONCLUSIONS: Bmal1 controls diurnal rhythms in expression and activity of intestinal CES1. Our findings have implications for understanding the crosstalk between circadian clock and xenobiotic metabolism in the intestine.

Proton solvent-controllable synthesis of manganese oxalate anode material for lithium-ion batteries.

Manganese oxalates with different structures and morphologies were prepared by the precipitation method in a mixture of dimethyl sulfoxide (DMSO) and proton solvents. The proton solvents play a key role in determining the structures and morphologies of manganese oxalate. Monoclinic MnC2O4·2H2O microrods are prepared in H2O-DMSO, while MnC2O4·H2O nanorods and nanosheets with low crystallinity are synthesized in ethylene glycol-DMSO and ethanol-DMSO, respectively. The corresponding dehydrated products are mesoporous MnC2O4 microrods, nanorods, and nanosheets, respectively. When used as anode material for Li-ion batteries, mesoporous MnC2O4 microrods, nanorods, and nanosheets deliver a capacity of 800, 838, and 548 mA h g-1 after 120 cycles at 8C, respectively. Even when charged/discharged at 20C, mesoporous MnC2O4 nanorods still provide a reversible capacity of 647 mA h g-1 after 600 cycles, exhibiting better rater performance and cycling stability. The electrochemical performance is greatly influenced by the synergistic effect of surface area, morphology, and size. Therefore, the mesoporous MnC2O4 nanorods are a promising anode material for Li-ion batteries due to their good cycle stability and rate performance.

Iron overload induces G1 phase arrest and autophagy in murine preosteoblast cells.

This study aimed to investigate the cell cycle arrest and autophagy induced by iron overload in MC3T3-E1 cells. MC3T3-E1 cells were cultured in different concentrations of ferric ammonium citrate (FAC), and Perls' Prussian blue reaction was used to detect the iron levels of the cells. CCK-8 assays were used to detect the growth of MC3T3-E1. The level of reactive oxygen species (ROS) within cells was investigated with DCFH-DA. PI staining was used to analyze the cell cycle distribution of MC3T3-E1 cells. Finally, the expression levels of cell cycle related proteins, autophagy related proteins, AKT, p38 MAPK, Stat3, and their downstream proteins were detected with Western blot assays. The results showed that the iron levels of MC3T3-E1 cells increased with increasing concentrations of FAC. High levels of ferric ion inhibited proliferation of MC3T3-E1 cells and increased their ROS levels. Additionally, iron overload induced G1arrest in MC3T3-E1 cells and down-regulated the expression of Cyclin D1 , Cyclin D3 , CDK2, CDK4 and CDK6, but up-regulated p27 Kip1. In addition, the expression levels of Beclin-1 and LC3 II increased, but that of p62 decreased. Further experiments showed that the phosphorylation of AKT and its downstream proteins p-GSK-3ß(Ser9) and p-mTOR (Ser2448) were decreased. The levels of p-p38 and p53 were up-regulated while those of cdc25A and p-ERK 1/2 were down-regulated. Phosphorylation of Stat3 and its downstream proteins was all decreased. These results show that iron overload generates ROS, blocks the PI3K/AKT and Jak/Stat3 signal pathways, and activates p38 MAPK, subsequently inducing G1 arrest and autophagy in MC3T3-E1 cells.

Customized Lingual Orthodontic Treatment of Skeletal Class II Malocclusion with Bilateral Maxillary First Premolar Extraction: Case Report.

This report describes the orthodontic camouflage treatment for a 52-year-old Chinese man using eBrace customized lingual appliance with bilateral maxillary first premolar extraction. The treatment results showed that using the eBrace customized lingual appliance can achieve expected effects and has a high level of safety for periodontal health.

Applications of cone-beam computed tomography to assess the effects of labial crown morphologies and collum angles on torque for maxillary anterior teeth.

INTRODUCTION: Currently, cone-beam computed tomography (CBCT) has been widely used because of its capacity to evaluate the anatomic structures of the maxilla, mandible, and teeth in 3 dimensions. However, articles about the use of CBCT to evaluate the relationships between the morphology of individual teeth and torque expression remain rare. In this study, we aimed to determine the influence of labial crown morphologies and collum angles on torque for maxillary anterior teeth using CBCT. METHODS: A total of 206 extracted maxillary anterior teeth were selected to establish scanning models using dental wax, and they were scanned by CBCT. Three-dimensionally reconstructed images and median sagittal sections of the teeth were digitized and analyzed with AutoCAD software (Autodesk, San Rafael, Calif). The angle α, formed by the intersection of the tangent at a certain vertical height on the labial surface from the incisal edge with the crown long axis, and the collum angle, were measured. RESULTS: The variations in angle α at different heights from the incisal edge for the same type of tooth were statistically significantly different (P <0.001). Moreover, the variations between collum angles and 0° for any type of maxillary anterior tooth were statistically significant (P <0.01). CONCLUSIONS: This study suggested that there are great differences in labial crown morphologies and collum angles for maxillary anterior teeth between persons, indicating that the morphologies of these teeth do play important roles in torque variations.