Ozone oil promotes wound healing via increasing miR-21-5p-mediated inhibition of RASA1.

Skin wound is a very common type of injury and the healing process greatly affects the life quality of individuals. Ozone has been shown beneficial to wound healing with unclear mechanisms. Here, we tested the effect of ozone oil (OZ) on wound healing and investigated the underlying mechanisms. Mouse skin wound model and Masson staining were used to evaluate the effect of OZ on wound healing. Primary fibroblast culture was employed to assess the functions of OZ, miR-21-5p, and RASA1. QRT-PCR and western blot were used to determine expression levels of miR-21-5p, RASA1, α-SMA, and collagen I. CCK-8 assay and scratch wound healing assay were used to measure viability and migration of fibroblasts. Dual luciferase activity assay was performed to validate miR-21-5p/RASA1 interaction. OZ accelerated wound healing in mice and promoted proliferation and migration abilities of fibroblasts. miR-21-5p was increased while RASA1 was reduced during the wound healing and OZ treatment augmented those changes, as well as increased levels of α-SMA and collagen I. Knockdown of miR-21-5p suppressed those effects of OZ on fibroblasts. Furthermore, miR-21-5p directly targeted RASA1 mRNA and negatively regulated its expression. Overexpression of RASA1 inhibited fibroblast proliferation and migration as well as diminished α-SMA and collagen I protein expression. Additionally, RASA1 overexpression blocked the promotion of miR-21-5p overexpression on fibroblast viability and migration. In vivo, miR-21-5p facilitated wound healing while overexpression of RASA1 reversed the effect. OZ promoted wound healing by enhancing miR-21-5p-mediated RASA1 inhibition to increase fibroblast proliferation and migration.

Silencing of microRNA-27a facilitates autophagy and apoptosis of melanoma cells through the activation of the SYK-dependent mTOR signaling pathway.

Melanoma is considered as an aggressive neoplastic transformation and featured with high metastatic potential. Although some studies have provided targets for novel therapeutic interventions, clinical development of targeted drugs for melanoma still remains obscure. Therefore, this study aims to identify the role of microRNA-27a (miR-27a) in autophagy and apoptosis of melanoma cells in regulating spleen tyrosine kinase (SYK)-mediated the mammalian target of rapamycin (mTOR) signaling pathway. A microarray-based analysis was made to screen differentially expressed genes and predict target miRNA. Melanoma specimens were collected with pigmented nevus as a control. Melanoma cell line Mel-RM was treated with miR-27a inhibitor or pcDNA-SYK to prove their effects on autophagy and apoptosis of melanoma cells. The volume change and tumor mass of nude mice in each group were detected by the tumorigenesis assay. Microarray-based analysis results showed that SYK was lowly expressed in melanoma cells and may be regulated by miR-27a. Besides, miR-27a expression was increased whereas SYK expression was decreased in melanoma tissues. Meanwhile, miR-27a was positively correlated with tumor stage and lymph node metastasis of melanoma tissues. Furthermore, miR-27a targeted SYK and silencing of miR-27a or overexpression of SYK cells promoted autophagy and apoptosis of melanoma cells and reduced their tumorigenic ability in vivo. In conclusion, this study proves that silencing of miR-27a facilitates autophagy and apoptosis of melanoma cells by upregulating SYK expression and activating the mTOR signaling pathway. The finding offers new ideas for the clinical development of melanoma.

Bearing Capacity Performance and Optimal Design of a Novel Aluminum Alloy Arch Gusset Joint.

Current research on aluminum alloy gusset joints has neglected the influences of the angle between members and the curvature of the joint plate on joint performance. This study introduces the concept of the planar angle and establishes 16 joint models using ABAQUS finite element software with parameters such as the planar angle, arch angles, joint plate thickness, web thickness, and flange thickness. The load-bearing capacity of the novel aluminum alloy arch gusset joint is theoretically analyzed, and the concepts of strong and weak axes are proposed. The failure modes and significance of different parameters regarding the bearing capacity and initial stiffness of the joint under various parameters are summarized. The results indicate that the planar and arch angles significantly affect the bearing capacity, stiffness, and failure mode of the joint.

LINC0638 lncRNA is involved in the local recurrence of melanoma following surgical resection.

Melanoma is a rare malignancy in China and the treatment outcomes are generally satisfactory. However, postoperative recurrence can be life-threatening. The current study aimed to investigate the involvement of long intergenic non-protein coding RNA 1638 (LINC01638) long non-coding RNA (lncRNA) in the recurrence of melanoma. Expression of LINC01638 lncRNA in skin biopsies and plasma of patients with melanoma, patients with benign skin lesions and healthy controls was detected by reverse transcription-quantitative polymerase chain reaction. Diagnostic values of LINC01638 lncRNA for melanoma were analyzed by receiver operating characteristic curve analysis. The association between LINC01638 lncRNA and clinicopathological data of patients with melanoma was analyzed by χ2 test. All patients were followed up for five years to record recurrence. LINC01638 lncRNA expression vectors and shRNAs were transfected into human melanoma cell lines and the effects of LINC01638 lncRNA overexpression and knockdown on cell proliferation were analyzed by cell counting kit-8 assay. LINC01638 lncRNA was significantly upregulated in patients with melanoma compared with the other two groups of patients, and upregulation of LINC01638 lncRNA distinguished patients with melanoma from patients with benign skin lesions and healthy controls. LINC01638 lncRNA expression was significantly associated with tumor size but not with other patient clinical data. Plasma levels of LINC01638 lncRNA were further increased during follow-up in patients with local recurrence but not in patients without recurrence. LINC01638 lncRNA overexpression promoted, while knockdown inhibited proliferation of cells of melanoma cell lines, C32 and SK-MEL-28, in vitro. The upregulation of LINC01638 lncRNA was likely associated with the local recurrence of melanoma following surgical resection.

Ozone oil promotes wound healing by increasing the migration of fibroblasts via PI3K/Akt/mTOR signaling pathway.

Skin injury affects millions of people via the uncontrolled inflammation and infection. Many cellular components including fibroblasts and signaling pathways such as transforming growth factor-ß (TGF-ß) were activated to facilitate the wound healing to repair injured tissues. C57BL/6 female mice were divided into control and ozone oil treated groups. Excisional wounds were made on the dorsal skin and the fibroblasts were isolated from granulation tissues. The skin injured mouse model revealed that ozone oil could significantly decrease the wound area and accelerate wound healing compared with control group. QPCR and Western blotting assays showed that ozone oil up-regulated collagen I, α-SMA, and TGF-ß1 mRNA and protein levels in fibroblasts. Wound healing assay demonstrated that ozone oil could increase the migration of fibroblasts. Western blotting assay demonstrated that ozone oil increased the epithelial-mesenchymal transition (EMT) process in fibroblasts via up-regulating fibronectin, vimentin, N-cadherin, MMP-2, MMP-9, insulin-like growth factor binding protein (IGFBP)-3, IGFBP5, and IGFBP6, and decreasing epithelial protein E-cadherin and cellular senescence marker p16 expression. Mechanistically, Western blotting assay revealed that ozone oil increased the phosphorylation of PI3K, Akt, and mTOR to regulate the EMT process, while inhibition of PI3K reversed this effect of ozone oil. At last, the results from Cytometric Bead Array (CBA) demonstrated ozone oil significantly decreased the inflammation in fibroblasts. Our results demonstrated that ozone oil facilitated the wound healing via increasing fibroblast migration and EMT process via PI3K/Akt/mTOR signaling pathway in vivo and in vitro The cellular and molecular mechanisms we found here may provide new therapeutic targets for the treatment of skin injury.

Protective effect of resveratrol against oxidative damage of UVA irradiated HaCaT cells.

OBJECTIVE: To observe the photoprotective effect and possible mechanisms of resveratrol for ultraviolet A (UVA) irradiated HaCaT cells. METHODS: HaCaT cells under UVA irradiation with 5J/cm(2) were interfered with 0.01 mmol/L and 0.1 mmol/L resveratrol. The testing objects were divided into a control and a UVA irradiation group, and then we detected the proliferation capacity with methylthiazdyl tetrazolium (MTT) and superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activity, content of maleic dialdehyde (MDA) with hydroxylamine, colorimetric, thiobarbituric acid (TBA) methods. The ultrastructure was observed under electron microscope. RESULTS: Resveratrol could enhance the proliferation activity, SOD, GSH-Px activity of HaCaT cells under UVA irradiation, decrease the content of MDA in dose-dependent manner (P<0.05). The electron microscope revealed that resveratrol could relieve the injury of HaCaT cells' ultrastructure. CONCLUSION: Resveratrol can relieve the inhibition to HaCaT cell proliferation,injury of their ultrastructure and oxidation by UVA irradiation. The protection is dose-dependent. That resveratrol raises the oxidase activity and clears the oxyradical may account for these results.