PD-L1 Inhibits T Cell-Induced Cytokines and Hyaluronan Expression via the CD40-CD40L Pathway in Orbital Fibroblasts From Patients With Thyroid Associated Ophthalmopathy.

Thyroid associated ophthalmopathy (TAO), characterized by T cell infiltration and orbital fibroblast activation, is an organ-specific autoimmune disease which is still short of effective and safety therapeutic drugs. The PD-1/PD-L1 pathway has been reported hindering the progression of Graves' disease to some extent by inhibiting T cell activity, and tumor therapy with a PD-1 inhibitor caused some adverse effects similar to the symptoms of TAO. These findings suggest that the PD-1/PD-L1 pathway may be associated with the pathogenesis of TAO. However, it remains unknown whether the PD-1/PD-L1 pathway is involved in orbital fibroblast activation. Here, we show that orbital fibroblasts from patients with TAO do not express PD-L1. Based on in vitro OF-T cell co-culture system, exogenous PD-L1 weakens T cell-induced orbital fibroblast activation by inhibiting T cell activity, resulting in reduced production of sICAM-1, IL-6, IL-8, and hyaluronan. Additionally, exogenous PD-L1 treatment also inhibits the expression of CD40 and the phosphorylation levels of MAPK and NF-κB pathways in orbital fibroblasts of the OF-T cell co-culture system. Knocking down CD40 with CD40 siRNA or down-regulating the phosphorylation levels of MAPK and NF-κB pathways with SB203580, PD98059, SP600125, and PDTC can both reduce the expression of these cytokines and hyaluronan. Our study demonstrates that the orbital immune tolerance deficiency caused by the lack of PD-L1 in orbital fibroblasts may be one of the causes for the active orbital inflammation in TAO patients, and the utilization of exogenous PD-L1 to reconstruct the orbital immune tolerance microenvironment may be a potential treatment strategy for TAO.

Plasma miR-6089 as potential diagnostic biomarker for retinoblastoma.

OBJECTIVE: The purpose of this study was to screen target miRNA related to RB and explore the expression levels of target miRNA in RB and its potential value of diagnosis. METHODS: The Affymetrix GeneChip miRNA 4.0 Array was used to screen the differential miRNAs in the plasma of 5 RB patients before and after intravenous chemotherapy, and the most significant down-regulated miRNA was selected for target miRNA. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) is used to verify the expression levels of plasma target miRNA in 30 RB patients. Then, qRT-PCR was performed to further verify the expression of target miRNA in plasma of RB patients and RB tumor tissues. Finally, receiver-operating-characteristic (ROC) curve and the area under the ROC curve (AUC) were used to evaluate the diagnostic power of plasma target miRNA. RESULTS: The miRNA Array obtain 8 core miRNAs, 1 up-regulated and 7 down-regulated, of which miR-6089 was the most significantly down-regulated. Plasma miR-6089 levels were significantly up-regulated in RB patients. Besides, in RB tumor tissues, miR-6089 levels were also obviously up-regulated. After intravenous chemotherapy, the expression of plasma miR-6089 was significantly decreased. Furthermore, ROC curve analysis showed that miR-6089 in the plasma had a good sensitivity and specificity for distinguishing RB from the healthy control group. CONCLUSIONS: MiR-6089 may be considered as a novel potential diagnostic biomarker for RB. TRIAL REGISTRATION NUMBER: ChiCTR2000040154; date of registration: 2020/11/22; retrospectively registered.

An Environmental Control Experiment for Contamination of the Production and Storage of 20% Autologous Serum Eye Drops.

Purpose: To investigate the sterility of autologous serum eye drops used for ocular surface diseases. Methods: A total of 100 patients were enrolled. The serum was prepared as follows: 20% serum (20% S), 20% serum with dexamethasone (0.02 mg/ml) (20% S + Dex), and 20% serum with levofloxacin (0.1 mg/ml) (20% S + Lev). Serum samples were collected for normal microbial cultivation at 1, 3, 5, 7, 10, 14, 21, and 28 days. The last samples were also assessed on the 28th day by airtight microbial cultivation. Results: A total of 2400 samples were cultured, and the bacterial contamination rates of 20% S, 20% S+ Dex, and 20% S + Lev group were 4.75%, 3.38%, and 0.88%, respectively, for normal microbial cultivation. There was no significant difference in bacterial contamination among the three groups with times (P = .502). Bacterial contamination of the 20% S + Lev group showed a significant difference in comparison with the 20% S or 20% S + Dex group (P < .05) in two culture methods; however, no significant difference was found between the 20% S and 20% S + Dex group (P = .208). There were two samples positive for fungi in the 20% S and 20% S + Dex group and three samples in the 20% S + Lev group in normal cultivation during 28 days. None of the samples was positive with fungi in airtight cultivation on the 28th day. There was also less bacterial contamination in airtight cultivation than in normal cultivation for the three groups on the 28th day. Conclusions: Our study shows that 20% autologous serum drops can be safely prepared and stored at 4°C in an open system under a strict protocol for at least 28 days, and antimicrobial agents could reduce the risk of contamination.