LncRNA MALAT1 regulates proliferation and apoptosis of vascular smooth muscle cells by targeting miRNA-124-3p/PPARα axis.

OBJECTIVE: To uncover the involvement of long non-coding RNA (lncRNA) MALAT1 in the proliferation and apoptosis of vascular smooth muscle cells (VSMCs), and the underlying mechanism. MATERIALS AND METHODS: Relative levels of MALAT1, microRNA-124-3p (miRNA-124-3p) and peroxisome proliferator-activated receptor alpha (PPARα) in VSMCs treated with different doses of oxidized low-density lipoprotein (ox-LDL) for different time points were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Proliferative and apoptotic changes of VSMCs overexpressing MALAT1 were assessed. Subcellular distribution of MALAT1 was analyzed. The potential binding among MALAT1, miRNA-124-3p and PPARα was determined by dual-luciferase reporter gene assay, and their interaction was determined as well. Finally, the influences of MALAT1/miRNA-124-3p/PPARα regulatory loop on the proliferative and apoptotic abilities of VSMCs were examined. RESULTS: MALAT1 and PPARα were dose-dependently downregulated in ox-LDL-treated VSMCs, whereas miRNA-124-3p was gradually upregulated. Overexpression of MALAT1 attenuated viability and induced apoptosis in ox-LDL-treated VSMCs. Moreover, MALAT1 was mainly distributed in the nucleus. Dual-luciferase reporter gene assay verified that MALAT1 could sponge miRNA-124-3p, and moreover, PPARα was the direct target of miRNA-124-3p. MALAT1 negatively regulated miRNA-124-3p level and miRNA-124-3p negatively regulated PPARα level as well. Finally, MALAT1/miRNA-124-3p/PPARα regulatory loop was identified to regulate the viability and apoptosis of ox-LDL-treated VSMCs. CONCLUSIONS: LncRNA MALAT1 mediates proliferation and apoptosis of VSMCs by sponging miRNA-124-3p to positively regulate PPARα level.

Folliculosebaceous cystic hamartoma arising within a port-wine stain.

A 34-year-old woman presented with 2-year history of a dome-shaped papule on a well-circumscribed, thickened, port-wine stain on the left side of the chin. Squeezing on the port-wine-stain plaque revealed many comedos within dilated follicular orifices. The papule was excised and submitted for histological examination. Histopathological study showed a lobular neoplasm, comprising dilated, cystic pilosebaceous structures surrounded by fibrous stroma, bearing the characteristics of folliculosebaceous cystic hamartoma. The reported case shows that, in addition to the vascular nature, both ectodermal and mesenchymal abnormalities may be involved in port-wine stains.

Clinical experience in managing Fusarium solani keratitis.

Fusarium solani keratitis is a rare ocular infectious disease. The clinical characteristics and treatment methods of 18 patients with culture proven F. solani keratitis between July 1997 and December 2003 and with a follow-up period of more than 4 months were analysed retrospectively. The patients were divided into two groups based on the severity of keratitis. Group A (n = 13) displayed non-severe keratitis and were treated with debridement, lamellar keratectomy and antifungal medication. Group B (n = 5) displayed severe keratomycosis and were treated with lamellar keratectomy combined with amniotic membrane transplantation (AMT) and antifungal medication. In group A, wound healing did not interfere with the integrity of the anterior chamber. The mean re-epithelialisation time was 12.67 days (range: 5-21 days). All patients were free of major immediate postoperative complications. In group B, AMT preserved the anterior chamber integrity in two cases, but failed to do so in the other three cases. Therapeutic patch grafts were required in these three cases. Non-severe F. solani keratitis is best treated with superficial keratectomy. Timely AMT combined with lamellar keratectomy appears to be an adjuvant therapy for severe keratomycosis and avoiding emergent therapeutic penetrating keratoplasty. However, AMT was effective in cases involving non-suppurative Fusarium keratitis.

Enhancement of topical 5-aminolaevulinic acid delivery by erbium:YAG laser and microdermabrasion: a comparison with iontophoresis and electroporation.

BACKGROUND: 5-aminolaevulinic acid (ALA) is used as a protoporphyrin IX-precursor for the photodynamic therapy of superficial skin cancer and cutaneous metastases of internal malignancies. However, the permeability of hydrophilic ALA across the skin is very low. OBJECTIVES AND METHODS: The objective of this study was to optimize and enhance the in vitro skin permeation of ALA by two resurfacing techniques: erbium:yttrium-aluminium-garnet (Erb:YAG) laser and microdermabrasion. Light microscopic changes in pig skin caused by these techniques were also compared. The electrically assisted methods, iontophoresis and electroporation, were also used to facilitate ALA permeation across laser- or microdermabrasion-treated skin. RESULTS: Among the modalities tested in this study the Erb:YAG laser showed the greatest enhancement of ALA permeation. The laser fluence was found to play an important role in controlling the drug flux, producing enhancement ratios from 4-fold to 246-fold relative to the control. The skin permeation of ALA across microdermabrasion-treated skin was approximately 5-15-fold higher than that across intact skin. Both the ablated effect of the stratum corneum (SC) and ALA flux were proportional to the treatment duration of microdermabrasion. The application of iontophoresis or electroporation alone also increased the ALA permeation by approximately 15-fold and 2-fold, respectively. The incorporation of iontophoresis or electroporation with the resurfacing techniques caused a profound synergistic effect on ALA permeation. CONCLUSIONS: This basic study has encouraged the further investigation of ALA permeation by laser or microdermabrasion.

Seeing the gene therapy: application of gene gun technique to transfect and decolour pigmented rat skin with human agouti signalling protein cDNA.

We developed a gene gun method for the transfer of human agouti signalling protein (ASP) cDNA to alter rat skin colour in vivo. Human ASP cDNA was cloned into a modified cytomegalovirus plasmid and delivered to the skin of Long-Evans rats by gene gun bombardment. Skin pigmentation, body weight and blood sugar of ASP cDNA-transfected rats were recorded against the control group, which were injected with plasmids encoding for green fluorescent protein. The treated skin showed lighter skin colour after 3 days of ASP gene transfection. This depigmentation effect was most prominent on day 14 and the skin gradually returned to its original pigmentation by day 28. Successful transfection of ASP gene in skin and hair follicles, as well as downregulation of melanocortin-1 receptor (MC1R) and tyrosinase expression upon treatment, was confirmed using immunohistochemistry and Western blot analysis. Body weight and blood sugar in the treated rats did not show statistically significant differences as compared to control groups. These observations demonstrate that gene transfer using the gene gun method can induce high cutaneous ASP production and facilitate a switch from dark to fair colour without systemic pleiotropic effects. Such a colour switch may be that ASP is acting in a paracrine fashion. In addition, this study verifies that ASP exerts its functions by acting as an independent ligand that downregulates the melanocyte MC1R and tyrosinase protein in an in vivo system. Our result offers new, interesting insights about the effect of ASP on pigmentation, providing a novel approach to study the molecular mechanisms underlying skin melanogenesis.