Study of serum exosome miRNA as a biomarker for early onset adult ouclar myastthenia gravis.

BACKGROUND: By extracting and sequencing miRNAs from serum exosomes of patients with early-onset ocular myasthenia gravis (OMG), generalized myasthenia gravis (GMG) and healthy controls, we screened differentially expressed miRNAs and explored the possibility as potential biomarkers for early-onset OMG. METHODS: Peripheral blood samples were collected from patients with early-onset OMG, early-onset GMG, and age-matched healthy subjects, with 6 samples in each group. All these patients were diagnosed as MG for the first time and did not undergo any treatment. Exosomes miRNAs were extracted from the serum and performed deep sequencing; the differentially expressed miRNAs were compared and analyzed between OMG, GMG, and healthy control groups using edgeR. The differential expression standard was set to | log2FC |>1, p < 0.05. Target prediction of mRNAs were performed using miRTarBase, TargetScan, and miRDB databases, and a protein-protein interaction (PPI) network was constructed subsequently. The miRNAs with a significant difference were validated using RT-qPCR (10 early-onset OMG patients, 10 early-onset GMG patients and 10 age-sex-matched healthy subjects), and the value of the area under the ROC curve (AUC) was used to assess the diagnostic accuracy and evaluate clinical prognostic value. RESULTS: In total, one upregulated (miR-130a-3p) miRNA was obtained through the upregulated intersection between control vs OMG and OMG vs GMG; four downregulated (miR-4712-3p; miR-6752-5p; miR-320d; miR-3614-3p) miRNAs were obtained through the downregulated intersection between control vs OMG and OMG vs GMG. A total of 408 target genes were predicted for the five differentially expressed miRNAs. The mTOR signaling pathway and Rap1 signaling pathway were significantly enriched based on the enrichment results. RT-qPCR findings revealed that for the OMG, the expression of miR-320d, miR-4712-3p and miR-3614-3p was markedly up-/down-regulated as compared to GMG and healthy control group. The AUC for the three miRNAs between OMG and healthy control groups were 0.78, 0.79 and 0.79 respectively; the AUC between OMG and GMG was 0.84. CONCLUSIONS: The present study identified three novel miRNAs as candidate biomarkers for early-onset OMG patients and it was expected to provide a possibility and a new orientation for serum exosomal miRNAs as OMG diagnostic biomarkers.

Single Center to Evaluate and Compare Anisometropic Amblyopia in Adults Using Blood Oxygenation Level-Dependent Functional Magnetic Resonance Imaging and Diffusion Kurtosis Imaging.

BACKGROUND Anisometropic amblyopia results from the unequal ability to focus between the right and left eyes. Blood oxygenation level-dependent functional magnetic resonance imaging (BOLD-fMRI) measures the proportion of oxygenated hemoglobin in specific areas. Diffusion kurtosis imaging (DKI) is a method of diffusion tensor imaging that estimates the skewed distribution of water diffusion probability. We aimed to evaluate and compare 11 adult patients with anisometropic amblyopia (AA) with 13 normally sighted healthy controls (HC) using BOLD-fMRI and DKI. MATERIAL AND METHODS Eleven adults with AA (age range 20-49; mean age 29.18±8.089) and 13 HC adults (age range 22-50; mean age 28.00±5.79) were recruited. DKI scanning used a single excitation echo-planar imaging sequence and a region of interest to obtain DKI parameters for optic radiation; the corpus callosum was manually placed, including mean kurtosis (MK), fractional anisotropic (FA), and mean diffusivity (MD) values; and BOLD data used a gradient-echo echo-planar imaging sequence. RESULTS The AA group had lower MK and FA of bilateral optic radiation than the HC group (P=0.008 and P=0.006, respectively) and higher MD than the HC group (P=0.005). The MK of the corpus callosum in the AA group was lower than that of HC group (P=0.012).Compared with the non-dominant eyes of the HC group, the amblyopic eyes in the AA group had less activation range and intensity in Brodmann areas 17, 18, and 19. CONCLUSIONS The combined use of DKI and BOLD-fMRI detected microstructural changes associated with local visual pathways and identified damage to the visual cortex in patients with amblyopia.

Crosslinked Decellularized Porcine Pericardium as a Substrate for Conjunctival Reconstruction.

Seeking for suitable conjunctival reconstruction substitutes to overcome the limitations of current substitutes, such as amniotic membrane, is urgent. Decellularized tissues have become a promising strategy for tissue engineering. In this study, we prepared decellularized porcine pericardium (DPP) scaffolds by the phospholipase A2 method and crosslinked them with aspartic acid (Asp) and human endothelial growth factor (hEGF) to enhance biological performance on the DPP, obtaining DPP-Asp-hEGF scaffolds. In vitro DPP showed lower apoptosis, highly desirable, well preservation of extracellular matrix components, and favorable macro-microstructure, which was confirmed by histology, immunofluorescence, electron microscopy, collagen and DNA quantification, and cytotoxicity assay, compared to the native porcine pericardium (NPP). The crosslinked efficacy of the DPP-Asp-hEGF was furtherer verified by in vitro experiments with Fourier transform infrared (FTIR) and X-ray diffraction (XRD). Through animal models of conjunctiva defect model, the DPP-Asp-hEGF revealed a closed, multilayer epithelium with an equal amount of goblet cells and no indication for conjunctival scarring after 28 days, compared to amniotic membrane (AM) groups and sham groups. These results suggested that DPP-Asp-hEGF can offer a good conjunctival reconstructive substitute both in structure and in function.

Generation of an induced pluripotent stem cell line (SYSUSCi001-A) from a congenital cataract patient carrying heterozygous mutations in BFSP1 and RHO.

The human induced pluripotent stem cell (hiPSC) line was generated from peripheral blood mononuclear cells (PBMCs) isolated from a 1-year-old boy who suffering from congenital cataract (CC), carrying heterozygous mutations in BFSP1 and RHO. PBMCs from this patient were reprogrammed into hiPSCs using non-integrative Sendai viral vectors expressing OCT4, SOX2, KLF4 and C-MYC. CC-hiPSCs had normal karyotype and showed pluripotency both in vitro and in vivo. The CC-hiPSCs would supply an important cell model for studying the pathogenesis of CC.

Targeting platelet-derived growth factor receptor ß inhibits the proliferation and motility of human pterygial fibroblasts.

Background: Pterygium, a common eye disease with high postoperative recurrence, lacks effective therapeutic strategies. Therefore, it's urgent to identify specific targets to develop rationally targeted molecular drugs for the pterygial therapy. Methods: The cell proliferation and motility were studied in both the primary human pterygial fibroblasts (hPFs) and an ex vivo pterygium model. hPFs transfected with the pCMV3-PDGFRB plasmid, PDGFRB siRNA and CRISPR/Cas9 system were used to determine the role of PDGFR-ß in pterygial fibroblasts functions. Western blotting, immunohistochemistry and immunofluorescence were performed to evaluate the expression of the key proteins. Results: PDGFR-ß expression in the pterygial stroma and primary hPFs was significantly higher than that in the conjunctiva and human conjunctival fibroblasts. PDGF-BB promoted the proliferation, migration and invasion of hPFs, which can be significantly suppressed by sunitinib via inhibition of the PDGFR-ß/extracellular signal-regulated kinase (ERK) pathway. In the ex vivo model, the knockout of PDGFRB and sunitinib treatment blocked the proliferation and motility of fibroblasts in the pterygial stroma via the suppression of PDGFR-ß/ERK pathway. Conclusion: This study demonstrates that PDGFR-ß may be a potential therapeutic target for pterygium, and inhibition of PDGFR-ß by sunitinib is a promising and effective approach for pterygium treatment.

Luteolin inhibits angiogenesis by blocking Gas6/Axl signaling pathway.

Growth arrest-specific protein 6 (Gas6) induces the activation of Axl receptor tyrosine kinase (Axl), which plays an important role in angiogenic processes, including proliferation, migration, invasion, tube formation and pericyte recruitment of endothelial cells. The inhibition of Gas6/Axl pathway has been demonstrated to be an effective anti-angiogenic therapy. Luteolin, which is a natural active flavonoid, has been reported to possess anti-angiogenic effects. However, the underlying mechanism of luteolin in anti-angiogenesis is not fully understood. Herein, we report that luteolin significantly inhibited the Gas6-induced proliferation, migration, invasion and tube formation of human microvascular endothelial cells (HMEC­1s) in vitro, and suppressed the Gas6-induced recruitment of human brain vascular pericytes (HBVPs) to the endothelial tubes. Luteolin also suppressed Gas6-induced microvessel sprouting in aortic ring assay and neovascularization in chick chorioallantoic membrane assay. The anti-angiogenic effect of luteolin may be associated with the inhibition of the Gas6/Axl pathway and its downstream phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathways. Taken together, the present study provides new evidence regarding an anti-angiogenic mechanism of luteolin, and supports the notion that the dietary intake of luteolin contributes to the treatment of pathological angiogenesis.

Intravitreal triamcinolone versus intravitreal bevacizumab for diabetic macular edema: a meta-analysis.

AIM: To compare the efficacy of the sole intravitreal triamcinolone (IVT) versus intravitreal bevacizumab (IVB) alone or IVB combined with IVT in the treatment of diabetic macular edema (DME). METHODS: Pertinent publications were identified through systematic searches of database and manually searching. Methodological quality of the literatures was valuated according to the Jadad Score. RevMan 5.1.0 was used to do the meta-analysis. Heterogeneity was determined and sensitivity was conducted. RESULTS: Six studies were ultimately included in the meta-analysis. The results of our analysis showed IVT had a statistically significant improvement in vision over the IVB at 1 month and 3 months (P<0.01). However, the reduction was not significant regarding central macular thickness (CMT) during the earlier (1 month and 3 months) follow-up period (P=0.12, P=0.41, respectively). At later visit (6 months), IVT had a significant decrease in CMT when compared to IVB (P<0.01) while no significant improvement in visual acuity (VA) was observed (P=0.14). The incidence of intraocular hypertension was 13/102 in IVT group during follow-up period while 0/103 in IVB group. The difference was significant (P<0.01). With regards to IVT versus IVB combined with IVT, there were no significant differences in CMT at 1 month (P=0.86) and 3 months (P=0.06). The incidence of intraocular hypertension was 6/67 in IVT group during follow-up period while 4/66 in IVB+IVT group. But the difference was not significant (P=0.53). CONCLUSION: Current evidence shows IVT is superior in improving VA at earlier follow-up (1 month and 3 months) and in reducing CMT at later follow-up (6 months) for DME. At other time, it is in favor of IVT treatment but there are no statistically significances. However, IVT has the side-effect of ocular hypertension. There is no adequate evidence of the benefit adding IVB to IVT in contrast to IVT alone.

Endogenous aspergillus endophthalmitis after kidney transplantation.

Endogenous aspergillus endophthalmitis(EAE) after kidney transplant is a rare but important clinical problem due to potentially devastating consequences. Early diagnosis of EAE, timely removal of affected vitreous by vitrectomy, proper anti-fungal treatment, all contributed to the successful control of the disease. Therapeutic success of EAE in post-transplant patients depends largely on prompt diagnosis. Definite diagnosis of EAE is based on positive culture results of vitreous specimen, while fundoscopy and B scan ultrasound may aid early diagnosis. In terms of anti-fungal medicine, amphotericin B has long been the first choice, but its systemic applicaiton has severe adverse reactions, especially for patients with impaired renal function. Herein, we report the treatment modality of EAE after kidney transplant with vitrectomy, systemic administration of micafungin plus voriconazole, topical application of fluconazol and amphotercin B.