Exosomes derived from tumor cells play a key role in tumor development. In the present study, we identified the bioactivity of exosomes released from WERI-Rb1 retinoblastoma cells in tumor angiogenesis, as well as the underlying mechanism, through biochemical methods and animal experiments. Our in vitro data showed that exosomes could be engulfed by human vesicle endothelial cells (HUVECs), significantly promote cell viability and induce an inflammatory response in HUVECs by increasing the expression of a series of related genes, such as IL-1, IL-6, IL-8, MCP-1, VCAM1, and ICAM1. Significant increases in migration and tube formation were also observed in the HUVECs incubated with exosomes. Moreover, experiments with a nude mouse xenotransplantation model showed that exosomes injected near tumors could be strongly absorbed by tumor cells. The numbers of endothelial cells and blood vessels were significantly increased in tumor tissues treated with exosomes compared to control tissues. Furthermore, to reveal the mechanism underlying exosome-mediated angiogenesis in retinoblastoma, we analyzed the levels of 12 microRNAs in the exosomes. Specifically, our data showed that miR-92a-3p was enriched in RB exosomes. Accordingly, miR-92a-3p was increased in the HUVECs incubated with these exosomes. After treatment with a miR-92a-3p inhibitor, the promoting effect of exosomes on the migration and tube formation of HUVECs was significantly abrogated. The expression of the angiogenesis-related genes mentioned above was markedly decreased in HUVECs. Similarly, treatment with a microRNA mimic also demonstrated that miR-92a-3p was involved in the angiogenesis of HUVECs. More importantly, bioinformatics analysis predicted that Krüppel-like factor 2 (KLF2), a member of the KLF family of zinc-finger transcription factors, might be an active target of miR-92a-3p. Notably, this prediction was confirmed both in vitro and in vivo. Thus, our work suggests that exosomal miR-92a-3p is involved in tumor angiogenesis and might be a promising therapeutic candidate for retinoblastoma.

Purpose: To assess and compare the dry-eye-associated parameters and corneal biomechanical status after small-incision lenticule extraction (SMILE) and femtosecond laser-assisted in situ keratomileusis (FS-LASIK). Methods: Sixty-five patients’ (128 eyes) were collected between November 2012 and April 2013. Thirty-five (69 eyes) underwent SMILE and thirty (59 eyes) underwent FS-LASIK. Visual acuity, manifest refraction, tear break-up time (TBUT), Schirmer’s test (ST), corneal sensitivity, Ocular Surface Disease Index (OSDI), corneal hysteresis (CH) and corneal resistance factor (CRF) were evaluated preoperatively and 1 day, 1 week, 1 month, 3 and 6 months postoperatively. Results: There was no significant difference in visual outcomes of SMILE and FS-LASIK (p = 0.208). TBUT, ST, CH and CRF decreased significantly after surgery in both groups (all p < 0.05); however, the decrease of TBUT, ST and CRF in the SMILE group was not as significant as that in the FS-LASIK group (all p < 0.05) and the change of CH value was comparable in both groups (p = 0.052).Corneal sensitivity was significantly decreased in the FS-LASIK group postoperatively (p < 0.05), but not significantly changed in the SMILE group (p > 0.05). OSDI scores were significantly increased after surgery for the two groups (all p < 0.05), and the scores were much higher in the FS-LASIK group than in the SMILE group (p = 0.000). Conclusion: The corneal biological healing of SMILE surgery was superior to FS-LASIK in the early period.

Purpose: To compare the functional optical zone (FOZ) and visual quality after small-incision lenticule extraction (SMILE) and femtosecond laser–assisted laser in situ keratomileusis (FS-LASIK) in correcting high myopia. Methods: Ninety-two eyes of 46 high myopic patients with the same programmed optical zone (POZ) received SMILE in one eye and FS-LASIK in the contralateral eye. FOZ was calculated using a refractive power method. The decentration, visual outcomes, wavefront aberrations, contrast sensitivity, and quality of vision (QoV) questionnaire were analyzed at 6 months postoperatively. Results: The postoperative visual and refractive outcomes were comparable between SMILE and FS-LASIK. The FOZ for SMILE (5.62 ± 0.31 mm) was larger than for FS-LASIK (5.35 ± 0.28 mm; P < 0.001). Moreover, the total decentration for SMILE (0.29 ± 0.14 mm) was greater than in FS-LASIK (0.22 ± 0.11 mm; P < 0.001). The induced change in spherical aberration was less for SMILE than for FS-LASIK (P < 0.001). There was better contrast sensitivity under the mesopic condition with glare for SMILE than for FS-LASIK (P = 0.024). However, no significant difference was found in QoV scores between the two groups. Conclusions: SMILE created a larger FOZ and greater decentration than FS-LASIK when the same POZ was designed in high myopia. Objective and subjective visual symptoms were comparable between SMILE and FS-LASIK. Translational Relevance: The differences in FOZ and decentration between SMILE and FS-LASIK have little effect on vision outcomes. Surgeons should consider the FOZ and decentration in surgical options in high myopia.

Activation of microglia plays a key role in the development of neovascular retinal diseases. Therefore, it is essential to reveal its pathophysiological and molecular mechanisms to interfere with disease progression. Here a publicly available single-cell RNA sequencing dataset is used to identify that intercellular communications from M1 microglia toward M0 microglia are increased in the retinal angiogenesis model via exosomes. Moreover, the results both in vitro and in vivo demonstrate that M1 microglia-derived exosomes promote the activation and enhance the proangiogenic ability of resting microglia. Based on miRNA sequencing of exosomes combined with gene interference, further results show that activated microglia-derived exosomes promoted microglial activation by transmitting polarized signals to M0 microglia via miR-155-5p. Subsequently, miR-155-5p suppresses Socs1 and activates the NFκB pathway, which ultimately causes the inflammatory cascade and amplifies the proangiogenic effect. In addition, upregulated Irf1 drives the expression of miR-155-5p in activated microglia, thus leading to an increase in the tendency of miR-155-5p to be encapsulated by exosomes. Thus, this study elucidates the critical role of intercellular communication among various types of microglia in the complex retinal microenvironment during angiogenesis, and contributes to the novel, targeted, and potential therapeutic strategies for clinical retinal neovascularization.

Blue light is a major component of visible light and digital displays. Over-exposure to blue light could cause retinal damage. However, the mechanism of its damage is not well defined. Here, we demonstrate that blue light (900 lux) impairs cell viability and induces cell apoptosis in retinal neurocytes in vitro. A DNA electrophoresis assay shows severe DNA damage in retinal neurocytes at 2 h after blue light treatment. γ-H2AX foci, a specific marker of DNA double-strand breaks (DSBs), is mainly located in the Map2-posotive neuron other than the glia cell. After assaying the expression level of proteins related to DNA repair, Mre11, Ligase IV and Ku80, we find that Ku80 is up-regulated in retinal neurocytes after blue light treatment. Interestingly, Ku80 is mainly expressed in glia fibrillary acidic protein (GFAP)-positive glia cells. Moreover, following blue light exposure in vivo, DNA DSBs are shown in the ganglion cell layer and only observed in Map2-positive cells. Furthermore, long-term blue light exposure significantly thinned the retina in vivo. Our findings demonstrate that blue light induces DNA DSBs in retinal neurons, and the damage is more pronounced compared to glia cells. Thus, this study provides new insights into the mechanisms of the effect of blue light on the retina.

Ocular inflammation is a common pathological condition of a series of retinal degenerative diseases. Tetramethylpyrazine (TMP), a Chinese herbal extraction, is widely used in the treatment of several ocular diseases in Eastern countries. However, the exact mechanisms correlating the vision protective effects of TMP have not been elucidated. Thus, this study aimed to investigate TMP's molecular targets in anti-inflammatory activity in endotoxin lipopolysaccharide (LPS)-induced retinal inflammation both in vitro and in vivo. The primary cultured retinal microglial cells were pretreated with TMP and then activated by LPS. We found pretreatment with TMP significantly inhibited LPS-induced upregulation of CD68, a marker of mononuclear microglia activation. The morphological changes induced by LPS were also inhibited by the TMP pretreatment. Moreover, Toll like receptor 4 (TLR4), phosphorylation of inhibitor of NF-κB alpha (p-IκB-α) and the translocation of nuclear factor kappa B p65 (NF-κB p65) were significantly downregulated in retinal microglial cells with TMP pretreatment, which indicated that TMP might suppress LPS-induced retinal microglial activation through TLR4/NF-κB signalling pathway. And these results were confirmed in vivo. Pretreatment with TMP inhibited microglial activation, migration and regeneration, especially in ganglion cell layer (GCL). In addition to the inhibition of TLR4, TMP significantly inhibited the translocation of NF-κB p-65 to the nucleus in vivo. The downstream genes of NF-κB, such as the pro-inflammatory cytokines interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β), were significantly downregulated by TMP pretreatment in the retina. Accordingly, the increased expression of cleaved caspase-3 and the decreased ratio of B-cell lymphoma-2 (Bcl-2) to Bcl-2 associated X Protein (Bax) were significantly attenuated by TMP. TUNEL assay also demonstrated that TMP exerted neuroprotective effects in the retina. Therefore, this study elucidated a novel mechanism that TMP inhibits retinal inflammation by inhibiting microglial activation via a TLR4/NF-κB signalling pathway.