Serum microRNA as a potential biomarker for the activity of thyroid eye disease.

The aim of this study is to characterize the microRNA (miRNA) expression signatures in patients with thyroid eye disease (TED) and identify miRNA biomarkers of disease activity. Total RNA was isolated from the sera of patients with TED (n = 10) and healthy controls (HCs, n = 5) using the miRNeasy Serum/Plasma Kit. The NanoString assay was used for the comprehensive analysis of 798 miRNA expression profiles. Analysis of specific miRNA signatures, mRNA target pathway analysis, and network analysis were performed. Patients with TED were divided into two groups according to disease activity: active and inactive TED groups. Differentially expressed circulating miRNAs were identified and tested using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) tests in the validation cohort. Among the 798 miRNAs analyzed, 173 differentially downregulated miRNAs were identified in TED patients compared to those in the HCs. Ten circulating miRNAs were differentially expressed between the active and inactive TED groups and regarded as candidate biomarkers for TED activity (one upregulated miRNA: miR-29c-3p; nine downregulated miRNAs: miR-4286, miR-941, miR-571, miR-129-2-3p, miR-484, miR-192-5p, miR-502-3p, miR-597-5p, and miR-296-3p). In the validation cohort, miR-484 and miR-192-5p showed significantly lower expression in the active TED group than in the inactive TED group. In conclusion, the expression levels of miR-484 and miR-192-5p differed significantly between the active and inactive TED groups, suggesting that these miRNAs could serve as circulating biomarkers of TED activity, however, these findings need to be validated in further studies.

Wnt signalling inhibits adipogenesis in orbital fibroblasts from patients with Graves' orbitopathy.

BACKGROUND/AIMS: To investigate the role of Wnt signalling in adipogenesis using an in vitro model of Graves' orbitopathy (GO). METHODS: Orbital fat was obtained from patients with GO and non-GO participants for primary orbital fibroblast (OF) culture. Expression levels of Wnt5a, Wnt10b, ß-catenin, phospho-ß-catenin and cyclin D1 were compared between GO and non-GO OFs. These expression levels were also determined during adipogenesis of GO and non-GO OFs. The effects of a stimulator and inhibitor of Wnt signalling on adipogenesis of GO and non-GO OFs were investigated. RESULTS: Western blotting analysis showed significant reductions in ß-catenin and cyclin D1 and significant enhancement of phospho-ß-catenin in OFs from patients with GO, compared with OFs from non-GO participants (p<0.05). Expression of Wnt5a, Wnt10b, ß-catenin and cyclin D1 in OFs was highest on day 0, and then gradually declined after induction of adipogenic differentiation. The expression levels of PPARγ, C/EBPα and C/EBPß were reduced in Wnt stimulator-treated OFs in a dose-dependent manner. Oil red O staining confirmed that a stimulator of Wnt inhibited adipogenesis in GO OFs. CONCLUSION: These results indicate that Wnt signalling inhibits adipogenesis in OFs from patients with GO and non-GO participants. Further studies are required to examine the potential of Wnt signalling as a target for therapeutic strategies.

Matrix Metalloproteinase-9 Point-of-Care Immunoassay after Dacryocystorhinostomy in Patients with Nasolacrimal Duct Obstruction.

PURPOSE: To compare matrix metalloproteinase-9 (MMP-9) point-of-care immunoassay (InflammaDry) results before and after dacryocystorhinostomy (DCR) in patients with nasolacrimal duct (NLD) obstruction. METHODS: Thirty-eight eyes of 38 patients who were diagnosed with unilateral NLD obstruction were treated with endoscopic DCR. Treatment response was monitored using InflammaDry test, tear meniscus height was measured by slit-lamp microscopy, and tear meniscus parameters (tear meniscus height, depth, and area) were measured by anterior segment optical coherence tomography at baseline and at 1 month after surgery. RESULTS: In 38 patients, the positive percentage of MMP-9 in diseased eyes was 100% (38 eyes), much higher than that in healthy fellow eyes (13.2%, 5 of 38 eyes) at baseline (p<0.001, χ2 test test). MMP-9 levels before and after surgery were significantly different (p<0.001), and MMP-9-positive eyes showed a significant improvement in MMP-9 grade after surgery (p< 0.0001; generalized McNemar test). Furthermore, other clinical parameters (tear meniscus area, tear meniscus height, and tear meniscus depth) showed a general improvement. CONCLUSION: The concentration of MMP-9 increased in the tears of patients' eyes with NLD obstruction compared to the contralateral healthy eyes. These abnormal findings showed a significant improvement after DCR. The improvement of postoperative MMP-9 expression might be because of the improvement of tear retention.

The retinal pigment epithelial response after retinal laser photocoagulation in diabetic mice.

To investigate the characteristics of regenerated retinal pigment epithelial (RPE) cells after retinal laser photocoagulation in diabetic mice. C57BL/6J mice were used to induce diabetes using intraperitoneal injection of streptozotocin. The proliferation of RPE cells after laser photocoagulation was determined using the 5-ethynyl-2'-deoxyuridine (EdU) assay in both diabetic and wild-type mice. The morphological changes of RPE cells were evaluated by using Voronoi diagram from immunostaining for ß-catenin. Characteristics of regenerated cells were evaluated by quantifying the mRNA and protein levels of RPE and epithelial-mesenchymal transition (EMT) markers. There were significantly less EdU-positive cells in laser-treated areas in diabetic mice than wild-type mice. Hexagonality was extensively lost in diabetic mice. Many EdU-positive cells were co-localized with Otx2-positive cells in the center of the laser-treated areas in wild-type mice, but only EdU-positive cells were widely distributed in diabetic mice. Quantitative analysis of mRNA and protein levels showed that the expression levels of RPE markers, Pax6, Mitf, and Otx2, were significantly decreased in RPE of diabetic mice compared with that of wild-type mice, whereas the expression levels of EMT markers, vimentin and fibronectin, were significantly increased. The proliferation and hexagonality of regenerating RPE cells were impaired after laser photocoagulation, and the regenerated RPE cells lost their original properties in diabetic mice. Further clinical research is needed to elucidate the RPE response after laser photocoagulation in diabetic patients.