Robust mitochondrial ROS production induces extensive double-strand breaks (DSBs) in telomeric DNA of effector T cells, where the DNA repair machinery is rapidly hyper-evoked to sense and ligate DSBs during the respiratory burst. However, whether effector T cells can exploit the DNA repair system to simultaneously potentiate their functional activation remains largely unknown, especially in the context of autoimmunity. Here, we demonstrate that non-homologous end joining (NHEJ), a predominant mechanism of DNA repair, is highly activated in pathogenic T helper 17 (pTh17) cells and exerts a previously unrecognized effect on shaping the pathogenic nature of pTh17s to trigger autoimmunity. Mechanistically, the perception of DSBs by KU proteins facilitates auto-phosphorylation of DNA-dependent protein kinase catalytic subunit (DNA-PKcs), which stabilizes RORγt to bind to the promoters of effector-gene loci, thus initiating the pTh17 effector program to induce autoimmunity. Using mass spectrometry and transcriptome analyses, we identified IER2 as a novel NHEJ factor that potentiates DNA-PKcs kinase activity in response to IL-23R stimulation, which is necessary for shaping Th17 pathogenicity. Therefore, targeting the immuno-pattern of the NHEJ system shows potential for the treatment of autoimmune diseases.

Autoimmune uveitis (AU) is a sight-threatening ocular autoimmune disorder that often manifests as retinal vasculitis. Increased neutrophil infiltration around retinal vessels has been reported during the progression of AU, while how they function is not fully recognized. Neutrophil extracellular traps (NETs), produced by activated neutrophils, have been suggested to be detrimental in autoimmune diseases. Here, we found that NETs were elevated in patients with active AU, and this was verified in an experimental AU (EAU) mouse model. Depletion of neutrophils or degradation of NETs with deoxyribonuclease-I (DNase I) could decrease CD4+ effector T cell (Teff) infiltration in retina and spleen to alleviate EAU. Moreover, we found that the expression of adhesion molecules, selectin, and antigen-presenting molecules was elevated in EAU retina and in retinal microvascular endothelial cells (RMECs) cocultured with NETs. The stimulated RMECs further facilitated CD4+ T cell adhesion, activation, and differentiation into Teffs. Mechanistically, NETs trigger RMEC activation by hastening cell senescence through the cyclic GMPAMP synthase (cGAS)/stimulator of interferon genes (STING) pathway. Slowing down senescence or inhibiting the cGAS/STING pathway in RMECs reduces the activation and differentiation of CD4+ T cells. These results suggest a deleterious role of NETs in AU. Targeting NETs would offer an effective therapeutic method.

Bruton’s tyrosine kinase (BTK), a key regulator of immune responses, has been extensively studied in B cells and innate immunocytes. Small-molecule BTK inhibitors (BTKi) have shown remarkable efficacy in multiple preclinical and clinical studies for autoimmune diseases via mediating the activation of B cells and innate immunocytes. Recent studies have demonstrated functional BTK expression in T cells, contributing to T cell activation and the pathogenesis of aplastic anemia. Whether BTK is involved in uveitis, the CD4+ T cell-mediated autoimmune disease, and whether it could be a therapeutic target remain unclear. We assessed BTK expression and phosphorylation levels in CD4+ T cells from Autoimmune uveitis (AU) patients and experimental autoimmune uveitis (EAU) mice. By administering BTKi to EAU mice, we evaluated its effects on ocular and systemic inflammation levels through fundus photography, histopathology, qPCR, and flow cytometry. In vitro, we assessed its impact on the Th1/Th17/Treg differentiation, and the pathogenicity of CD4+ T cells. Transcriptomic sequencing and flow cytometry were employed to explore the potential mechanism by which BTK regulates CD4+ T cells. BTK expression and activation were upregulated in CD4+ T cells from AU patients and EAU mice. BTK inhibition suppressed the polarization of Th17 and Th1 cells, but not Treg differentiation. BTK inhibition upregulated the PPAR-γ signaling, alleviating oxidative stress and inflammatory responses in CD4+ T cells, thereby reducing ocular and systemic inflammation in EAU mice. Additionally, BTKi exhibited significant anti-inflammatory effects in CD4+ T cells from AU patients. This study demonstrates the critical role of BTK in CD4+ T cell-driven AU. BTK inhibition ameliorates oxidative stress and inflammatory responses via PPAR-γ involved signaling in CD4+ T cells and suppresses their differentiation to Th1/Th17 to restore immune homeostasis, thereby mitigating AU. These findings identify BTK as a potential therapeutic target and provide a theoretical foundation for the clinical application of BTKi in AU.

Purpose:The purpose of this study was to explore the underlying mechanism that Th17-like T follicular helper cells (Tfh) orchestrated by STING signaling have a pathogenic role in experimental autoimmune uveitis (EAU). Methods:The differences of transcriptome and gene ontology (GO) pathway of Tfh between EAU and control mice were analyzed by single-cell RNA sequence (scRNA-seq) and bulk RNA sequence. Additionally, draining lymph nodes (DLNs) were extracted to verify the expression of IL-17A and IFN-γ in Tfh from EAU and control mice by flow cytometry. Then, the scRNA-seq and flow cytometry were used to explore the different proportion of Tfh between STING deficiency (Sting-/-) mice and wild type (WT) mice. In vitro, naïve CD4+ T cells were isolated from Sting-/- mice and WT mice to induce the Tfh under the induction condition. In addition, flow cytometry was used to detect the different induction ratio and the IL-17A expression between 2 groups of naïve CD4+ T cells. Results:Compared with control mice, marked increase of Tfh was observed in EAU, accompanied by elevated levels of Th1 and Th17 cells. Moreover, Th17-related genes, such as Rorc, Il22, Il23r, Il17a, and Il17f, and the corresponding GO pathways were upregulated in Tfh from EAU. The scRNA-seq showed that a higher proportion of Tfh was observed in the DLNs from Sting-/- mice than WT mice, which was verified by flow cytometry. When STING was knocked out, the Tfh was characterized with upregulated Th17-related phenotype in vivo, and there was a higher induction ratio of Tfh whose IL-17A expression was significantly increased in vitro. Notably, the STING expression of CD4+ T cells was downregulated in the EAU. STING-deficient EAU mice displayed more severe retinal inflammation, characterized by massive infiltration of CD4+ T cells, including Th1 and Th17 subsets. Importantly, treatment with a STING agonist alleviated inflammation of EAU. Conclusions:Th17-like Tfh cells play a pathogenic role in the EAU. STING deficiency promotes the differentiation and phenotypic transformation of Th17-like Tfh cells, exacerbating the inflammatory response in EAU. These findings highlight the potential of targeting STING to modulate Tfh cells as a therapeutic strategy for uveitis.

BACKGROUND:Autoimmune uveitis is a sight-threatening inflammatory disease of the retina. MicroRNA-142 (miR-142) has been implicated in its pathogenesis. This study aimed to elucidate the role of miR-142 in uveitis and its underlying mechanisms. METHODS:The expression of miR-142-3p was analyzed in peripheral blood mononuclear cells from uveitis patients and in experimental autoimmune uveitis (EAU) models. With EAU induction for 14 days, clinical and histopathological scores were graded to evaluate the retinal inflammation. To investigate the effects of miR-142 deficiency on uveitis development, the miR-142 knockout (miR-142-/-) mouse model was used. The miR-142-/- T cell phenotype and function were characterized using flow cytometry and single-cell sequencing for both in vivo and in vitro experiments. The Seahorse Analyzer, mitochondrial staining and electron microscope analysis were conducted to reveal the mitochondrial function and morphology. And then Luciferase Assays and Western-Blot analysis were used to explore the target of miR-142. RESULTS:We found that miR-142-3p was significantly up-regulated in uveitis and that its deletion in mice prevented EAU development. The T cell isolated from miR-142-/- mice lose its uveitogenic nature. T cell lacking miR-142 exhibited reduced numbers and attenuated pathogenicity in uveitis, characterized by decreased proliferation, increased apoptosis, and abnormal differentiation. Single-cell sequencing, energy metabolism analysis and flow cytometry analysis unveiled metabolic reprogramming in miR-142-/- T cells, with a distinct shift toward glycolysis and restrained oxidative phosphorylation. Further investigation revealed mitochondrial fission regulator 1 (MTFR1) as a direct target of miR-142. The over-expressed protein of MTFR1 in CD4+ T cells was found in miR-142-/- mice. CONCLUSIONS:Our findings highlight the indispensable role of miR-142 in maintaining T cell mitochondrial function. By modulating MTFR1, miR-142 orchestrates mitochondrial homeostasis, metabolic alterations, apoptosis susceptibility, and proliferation capacity in T cells, thereby influencing susceptibility to autoimmune uveitis.

Circadian rhythm plays a critical role in the progression of autoimmune diseases. While our previous study demonstrated the therapeutic effects of melatonin in experimental autoimmune uveitis, the involvement of circadian rhythm remained unclear. Using a light-induced circadian rhythm disruption model, we showed that disrupted circadian rhythms exacerbate autoimmune uveitis by impairing the stability and function of Treg cells. Mechanistically, we identified the core clock gene Per1, which is significantly reduced under circadian disruption, is essential for Treg cell metabolism and immunoregulatory function. This study underscores the pivotal role of circadian rhythm-related Treg cells in autoimmune disease progression.

Bacterial keratitis is one of the leading causes of blindness in the world. Frequent administration of antibiotic eye drops is the first-line treatment, which has difficulties in achieving favorable outcomes in some cases due to the emerging antibiotic resistance and low bioavailability. Antimicrobial peptides have been shown to overcome antibiotic resistance through diverse bactericidal mechanisms including antibiofilm effects, enabling them to be a promising strategy to combat resistance. Here, we report a novel antibacterial strategy with collagen nanosheets (CN) loaded with the cationic antimicrobial short peptide Tet213 (Tet213-CN). In this study, we demonstrated that Tet213-CN possessed excellent properties with good adherence, high oxygen permeability, transparency, biocompatibility, and showed no ocular irritation. In vitro and in vivo assays demonstrated that Tet213-CN had enhanced antibacterial and corneal epithelial healing effects. Furthermore, we observed that Tet213-CN treatment facilitated innate immune defense by promoting M1 macrophage polarization, increasing intracellular ROS generation and proinflammatory cytokine secretion, and enhancing phagocytosis of bacteria via the TLR2/MyD88/NF-κB signaling pathway. Most impressively, Tet213-CN achieved considerable outcomes in Pseudomonas aeruginosa keratitis models with a one-time application. Overall, Tet213-CN may serve as an ideal strategy in the management of bacterial keratitis, and shed new light onto the treatment against antibiotic-resistant bacteria. In addition, these results provide a conceptual framework for a novel macrophage-based strategy in the treatment of corneal infection diseases.

Abstract Background Thyroid eye disease (TED) is a vision-threatening autoimmune disorder. Orbital tissue fibrosis leading to intractable complications remains a troublesome issue in TED management. Exploration of novel therapeutic targets and agents to ameliorate tissue fibrosis is crucial for TED. Recent work suggests that Ca2+ signaling participates in tissue fibrosis. However, whether an alteration of Ca2+ signaling has a role in fibrogenesis during TED remains unclear. In this study, we aimed to investigate the role of Ca2+ signaling in the fibrogenesis process during TED and the potential therapeutic effects of a highly selective inhibitor of the L-type calcium channel (LTCC), nimodipine, through a TGF-β1 induced in vitro TED model. Methods Primary culture of orbital fibroblasts (OFs) were established from orbital adipose connective tissues of patients with TED and healthy control donors. Real-time quantitative polymerase chain reaction (RT-qPCR) and RNA sequencing were used to assess the genes expression associated with LTCC in OFs. Flow cytometry, RT-qPCR, 5-ethynyl-2′-deoxyuridine (EdU) proliferation assay, wound healing assay and Western blot (WB) were used to assess the intracellular Ca2+ response on TGF-β1 stimulation, and to evaluate the potential therapeutic effects of nimodipine in the TGF-β1 induced in vitro TED model. The roles of Ca2+/calmodulin-dependent protein kinase II (CaMKII) and signal transducer and activator of transcription 1 (STAT1) in fibrogenesis during TED were determined by immunohistochemistry, WB, flow cytometry and co-immunoprecipitation assay. Selective inhibitors were used to explore the downstream signaling pathways. Results LTCC inhibitor nimodipine blocked the TGF-β1 induced intracellular Ca2+ response and further reduced the expression of alpha-smooth muscle actin (α-SMA), collagen type I alpha 1 (Col1A1) and collagen type I alpha 2 (Col1A2) in OFs. Besides, nimodipine inhibited cell proliferation and migration of OFs. Moreover, our results provided evidence that activation of the CaMKII/STAT1 signaling pathway was involved in fibrogenesis during TED, and nimodipine inhibited the pro-fibrotic functions of OFs by down-regulating the CaMKII/STAT1 signaling pathway. Conclusions TGF-β1 induces an LTCC-mediated Ca2+ response, followed by activation of CaMKII/STAT1 signaling pathway, which promotes the pro-fibrotic functions of OFs and participates in fibrogenesis during TED. Nimodipine exerts potent anti-fibrotic benefits in vitro by suppressing the CaMKII/STAT1 signaling pathway. Our work deepens our understanding of the fibrogenesis process during TED and provides potential therapeutic targets and alternative candidate for TED.

PURPOSE. Experimental autoimmune uveitis (EAU) is a representative animal model of human uveitis. In this study, we investigated whether apolipoprotein A1 (APOA1) can alleviate EAU and explored its underlying mechanism. METHODS. Mice were immunized with interphotoreceptor retinoid-binding protein 1-20 and treated with APOA1 or vehicle. The retinas, draining lymph nodes (DLNs), and spleens were analyzed. Isolated T cells were used for proliferation, differentiation, and function assays in vitro. Selective inhibitors and pathway agonists were used to study signaling pathways. The effect of APOA1 on peripheral blood mononuclear cells (PBMCs) from uveitis patients was also examined. RESULTS. Administration of APOA1 ameliorated EAU. APOA1 suppressed pathogenic CD4+ T cell expansion in DLNs and spleen, and decreased the infiltration of effector T (Teff) cells into retina. APOA1 also inhibited T cell proliferation and T helper 1 cell differentiation in vitro and promoted regulatory T (Treg) cell differentiation. APOA1 restricted inflammatory cytokine production from lipopolysaccharide-stimulated PBMCs. Mechanistic studies revealed that the effect of APOA1 was mediated by scavenger receptor class B type I (SR-BI) and downstream signals including phosphatidylinositol 3-kinase/Protein kinase B (PKB, or Akt), p38 mitogen-activated protein kinase, and nuclear factor-kappa B. CONCLUSIONS. APOA1 ameliorates EAU by regulating the Teff/Treg partially through SR-BI. Our results suggest that APOA1 can be a therapeutic alternative for autoimmune uveitis.

Abstract Background Melatonin, an indoleamine produced by the pineal gland, plays a pivotal role in maintaining circadian rhythm homeostasis. Recently, the strong antioxidant and anti-inflammatory properties of melatonin have attracted attention of researchers. We evaluated the therapeutic efficacy of melatonin in experimental autoimmune uveitis (EAU), which is a representative animal model of human autoimmune uveitis. Methods EAU was induced in mice via immunization with the peptide interphotoreceptor retinoid binding protein 1–20 (IRBP1–20). Melatonin was then administered via intraperitoneal injection to induce protection against EAU. With EAU induction for 14 days, clinical and histopathological scores were graded to evaluate the disease progression. T lymphocytes accumulation and the expression of inflammatory cytokines in the retinas were assessed via flow cytometry and RT-PCR, respectively. T helper 1 (Th1), T helper 17 (Th17), and regulatory T (Treg) cells were detected via flow cytometry for both in vivo and in vitro experiments. Reactive-oxygen species (ROS) from CD4 + T cells was tested via flow cytometry. The expression of thioredoxin-interacting protein (TXNIP) and hypoxia-inducible factor 1 alpha (HIF-1α) proteins were quantified via western blot. Results Melatonin treatment resulted in notable attenuation of ocular inflammation in EAU mice, evidenced by decreasing optic disc edema, few signs of retinal vasculitis, and minimal retinal and choroidal infiltrates. Mechanistic studies revealed that melatonin restricted the proliferation of peripheral Th1 and Th17 cells by suppressing their transcription factors and potentiated Treg cells. In vitro studies corroborated that melatonin restrained the polarization of retina-specific T cells towards Th17 and Th1 cells in addition to enhancing the proportion of Treg cells. Pretreatment of retina-specific T cells with melatonin failed to induce EAU in naïve recipients. Furthermore, the ROS/ TXNIP/ HIF-1α pathway was shown to mediate the therapeutic effect of melatonin in EAU. Conclusions Melatonin regulates autoimmune T cells by restraining effector T cells and facilitating Treg generation, indicating that melatonin could be a hopeful treatment alternative for autoimmune uveitis.

PurposeInflammation triggers the activation of CD4+T cells and the breakdown of blood–retinal barrier, thus contributing to the pathology of experimental autoimmune uveitis (EAU). We explored the anti-inflammatory effect of hydroxychloroquine (HCQ) on EAU and the potential mechanisms active in T cells and retinal vascular endothelial cells (RVECs).MethodsC57BL/6J mice were immunized with interphotoreceptor retinoid binding protein 1-20 (IRBP1–20) to induce EAU and then treated with the vehicle or HCQ (100 mg/kg/day). On day 7, 14, 21, 30 and 60 after immunization, clinical scores were evaluated. On day 14, histopathological scores were assessed, and retinas, spleens, and lymph nodes were collected for quantitative polymerase chain reaction or flow cytometry analysis. RVEC dysfunction was induced by tumor necrosis factor α (TNF-α) stimulation. The expression of cytokines, chemokines, adhesion molecules, and lectin-like oxidized LDL receptor-1 (LOX-1)/nuclear factor κB (NF-κB) was measured in RVECs with or without HCQ.ResultsHCQ treatment protected mice from uveitis, evidenced by reduced expression of inflammatory factors, chemokines, and adhesion molecules in the retina. In systemic immune response, HCQ inhibited the activation of naïve CD4+T cells and frequencies of T effector cells, and promoted T regulatory cells. HCQ decreased IRBP1-20–specific T cell responses and proliferation of CD4+T cells in vitro. Further studies established that TNF-α induced RVECs to express inflammatory cytokines, chemokines, and adhesion molecules, whereas HCQ alleviated the alterations via the LOX-1/NF-κB pathways.ConclusionsHCQ alleviates EAU by regulating the Teff/Treg balance and ameliorating RVECs dysfunction via the LOX-1/NF-κB axis. HCQ may be a promising therapeutic candidate for uveitis.

Uveitis is one of the most common blindness-causing ocular disorders. Due to its complicated pathogenesis, the treatment of uveitis has been widely recognized as a challenge for ophthalmologists. Recently, the anti-inflammatory properties of the antibiotic Azithromycin (AZM) have been reported. However, the therapeutic effects of Azithromycin in experimental autoimmune uveitis (EAU), a representative model of human AU, have not been elucidated till date. We conducted this study to examine the therapeutic effects and potential mecha-nisms of Azithromycin in EAU. We observed that Azithromycin significantly attenuated retinal inflammation in EAU mice at day 14 after immunization along with a significantly decreased inflammatory cell infiltration and cytokine production in the retina. Furthermore, we observed that Azithromycin increased the number of regu-latory T cells (Treg) and decreased the number of effector T cells (Teff) in both the draining lymph nodes and spleen of EAU mice. Additionally, Azithromycin suppressed the proliferation and activation of CD4 + T cells, and induced the apoptosis of CD4 + CD44 + memory T and CD4 + CXCR3 + Th1 cells. Mechanistically, we proved that Azithromycin could regulate Teff/Treg balance by inhibiting the phosphorylation of S6 ribosomal protein, a downstream target of mammalian target of rapamycin (mTOR). Together, our findings revealed that Azi-thromycin alleviated EAU by regulating the Teff/Treg balance through the mTOR signaling pathway, suggesting that Azithromycin could be a promising therapeutic candidate for AU.

PURPOSE:Surgical excision is the standard treatment for pterygium. This study was conducted to evaluate the safety and efficacy of a novel technique using low-temperature plasma (LTP) for excision and hemostasis in pterygium surgery.METHODS:A prospective, comparative, and randomized clinical trial was conducted on 60 patients (60 eyes) undergoing pterygium excision with conjunctival autografts using fibrin glue. Patients were equally divided into the following 2 groups: a control group and a LTP group. Postoperative follow-up visits were scheduled on day 1, week 1, and months 1 and 3, and recurrence was evaluated at 1 year. Patients were examined for operative time, best corrected visual acuity, conjunctival autograft inflammation (CAI), graft stability (GS), pain, recurrence, and final appearance. Factors related to pterygium recurrence and final appearance were analyzed.RESULTS:Mean operative times were shorter in the LTP group (16.7 ± 3.4 min) than those in the control group (20.1 ± 4.7 min, P = 0.002). LTP eyes had milder CAI than control eyes at postoperative day 1 (P = 0.000) and week 1 (P = 0.000). Patients in the LTP group exhibited better GS (P = 0.01) and milder pain (P = 0.04) than those in the control group on day 1. Two control patients (6.7%) and no (0%) LTP patients experienced recurrence (P = 0.08). GS and CAI were the significant factors contributing to recurrence (GS: R = 0.425, P = 0.001; CAI: R = 0.309, P = 0.016).CONCLUSIONS:LTP to replace surgical blades and disposable cautery for ablation and hemostasis is safe and efficient for pterygium surgery, resulting in shorter operative time, milder inflammation, and better graft stability without increasing complication risk.

Corneal ulcer is a common ophthalmic symptom. Segmentation algorithms are needed to identify and quantify corneal ulcers from ocular staining images. Developments of such algorithms have been obstructed by a lack of high quality datasets (the ocular staining images and the corresponding gold-standard ulcer segmentation labels), especially for supervised learning based segmentation algorithms. In such context, we prepare a dataset containing 712 ocular staining images and the associated segmentation labels of flaky corneal ulcers. In addition to segmentation labels for flaky corneal ulcers, we also provide each image with three-fold class labels: firstly, each image has a label in terms of its general ulcer pattern; secondly, each image has a label in terms of its specific ulcer pattern; thirdly, each image has a label indicating its ulcer severity degree. This dataset not only provides an excellent opportunity for investigating the accuracy and reliability of different segmentation and classification algorithms for corneal ulcers, but also advances the development of new supervised learning based algorithms especially those in the deep learning framework.

Acellular porcine corneal stroma (APCS) has proven to be a promising alternative to traditional corneal grafts. This prospective case series was conducted to further investigate the healing characteristics of APCS following keratoplasty.

Purpose: To determine the prevalence, risk factors and microbial profiles of donor corneal contamination and its association with postoperative infection. Materials and Methods: 1348 hypothermic preserved donor corneas were screened during keratoplasty to assess the impacts of donor age, gender, cause of death and corneal preservation time on the contamination risk. The microbial spectrum and antibiotic sensitivity of causative microorganisms and the prognostic role of corneoscleral rim cultures were analyzed. Results: 111 donor corneas (8.2%) had positive microbial cultures, with 84 contaminated by bacteria, 25 by fungi and 2 by both. Acinetobacter baumannii complex (19.8%) and Candida spp. (9.0%) were the most commonly isolated bacteria and fungi, respectively. Two patients (1.8%) who received contaminated corneal buttons developed postoperative infections. Death due to cardiac disease led to more corneal contaminations than death due to brain disease (odds ratio (OR) = 2.59, P = .009). Longer preservation time was associated with a trend toward increasing contamination rate (from 8.3% to 15.0%). Moreover, fungal-contaminated corneas were preserved longer than bacterial-contaminated corneas (6.6 ± 4.5 versus 10.2 ± 5.4 days, P = .001). Corneas from donors who died from cardiac diseases and trauma showed the highest prevalence of bacterial (10.9%) and fungal (2.6%) contamination, respectively. Antibiotic sensitivity testing revealed that the third-generation fluoroquinolone levofloxacin had high rates of susceptibility to both gram-positive (G+) (60.0%) and gram-negative (G-) (44.6%) bacteria. Conclusions: The causes of donor corneal contamination are multifactorial. The antibiotic resistance rate of contaminating microbes seems to be increasing. Whether antibiotic usage in storage medium and postoperative prophylaxis should be updated accordingly warrants further investigation.

In this paper, we proposed and validated a novel and accurate automatic pipeline for extracting flaky corneal ulcer areas based on fluorescein staining images. We first used an existing semi-automatic approach to identify the cornea from each image. The ulcer area was then segmented within the cornea by employing a combination of techniques: 1) identify and modify the color information of reflective areas; 2) segment each image into a total of 1000 superpixels based on simple linear iterative clustering (SLIC); 3) employ support vector machine (SVM) to classify all superpixels into two classes; 4) erode and dilate to polish the ulcer segmentation results. The proposed pipeline has been validated on a total of 150 clinical images. Accurate segmentation results have been obtained, with the mean accuracy being 0.984, the mean Jaccard similarity coefficient being 0.871, and the Pearson correlation coefficient being 0.921 when compared with the manually-delineated gold standard. The proposed method was found to significantly outperform two classic segmentation algorithms (active contour and Otsu thresholding) in terms of segmenting corneal ulcers.

Purpose To investigate the expression and roles of type I and II interferons (IFNs) in fungal keratitis, as well as the therapeutic effects of tacrolimus (FK506) and voriconazole on this condition. Methods The mRNA and protein expression levels of type I (IFN-α/β) and II (IFN-γ) IFNs, as well as of related downstream inflammatory cytokines (interleukin (IL)-1α, IL-6, IL-12, and IL-17), were detected in macrophages, neutrophils, lymphocytes, and corneal epithelial cells (A6(1) cells) stimulated with zymosan (10 mg/ml) for 8 or 24 h. A fungal keratitis mouse model was generated through intrastromal injection of Aspergillus fumigatus, and the mice were then divided into four groups: group I, the PBS group; group II, the voriconazole group; group III, the FK506 group; and group IV, the voriconazole plus 0.05% FK506 group. Corneal damage was evaluated with clinical scoring and histological examination. In addition, the mRNA and protein expression levels of type I (IFN-α/β) and type II (IFN-γ) IFNs, as well as related inflammatory cytokines, were determined at different time points using quantitative real-time PCR (qRT-PCR) and western blotting. Results After zymosan stimulation of mouse neutrophils, lymphocytes, macrophages, and A6(1) cells, the IFN mRNA and protein expression levels were markedly increased until 24 h, peaking at 8 h (p<0.001). The mRNA and protein expression levels of inflammatory cytokines (IL-1α, IL-6, IL-12, and IL-17) were also upregulated after zymosan stimulation. Moreover, type I (IFN-α/β) and type II (IFN-γ) IFN expression levels were increased and positively correlated with the progression of fungal keratitis in vivo. FK506 administered with voriconazole reduced the pathological infiltration of inflammatory cells into the cornea and downregulated the expression levels of IFNs and related inflammatory cytokines. Conclusions In conclusion, this study demonstrated that type I and II IFN levels were markedly increased in fungal keratitis and that FK506 combined with voriconazole decreased the severity of fungal keratitis by suppressing type I and II IFNs and their related inflammatory responses.

Objective To investigate the characteristics of ocular surface inflammation index after pterygium excision by using Oculus Keratograph(R) 5M and Visual analogue scale and evaluated the effectiveness of antiinflammatory treatment.Methods A prospective case control study was performed.Eighteen patients (6 males and 12 females) who suffered from primary pterygium were recruited in Zhongshan Ophthalmic Center from June to September 2016.All patients were treated with monocular pterygium excision combined with amniotic membrane transplantation.Anti-inflammatory treatment was given after surgery,and the ocular inflammation index was evaluated at preoperative and 1st,3rd,7th,10th,30th and 60th day postoperative.The temporal conjunctival hyperemia index (TCHI) was assessed by Oculus Keratograph(R) 5M with a red eye index analysis software.Ocular symptom scores (OSS) and visual analogue scale (VAS) were used to analyze the subjective symptoms of the patients.Fluorescein staining was used to detect the epithelization of corneal and scleral wound.The best corrected visual acuity (BCVA),intraocular pressure and complications were evaluated in this study.This study was approved by the Medical Ethics Committee of Zhongshan Ophthalmic Center of Sun Yat-sen University (2016KYPJ024).All patients signed informed consent for clinical research.Results No drug-related ocular and systemic adverse events were found during the follow-up.Corneal epithelial defect was recovered on 10th day,and conjunctival epithelization was observed in sclera exposed area on 30th day.The BCVA on the 60th day was 0.12±0.17,which was significantly lower than 0.34±0.36 preoperatively (t =3.401,P =0.003).Compared with those before surgery,OSS and VAS were significantly increased on 1 st day (OSS:Z =-4.255,P =0.000;VAS:Z =-5.256,P =0.000).The OSS on 7th day was not significantly different from that before surgery (Z=-0.958,P=0.372).VAS decreased to baseline on 30th day.The OSS on 60th day after surgery was significantly lower than that before surgery (Z =-2.397,P =0.037).TCHI was higher than 1.2 preoperatively,and increased to the highest on 1 st day after surgery,with significant difference between them (t=-6.620,P=0.000).The TCHI decreased to baseline on 7th day,no significant difference were obtained when compared with preoperative TCHI (t =-1.050,P =0.310),and TCHI on 60th day after surgery was lower than that before surgery,with significant difference between them (t =2.758,P =0.020).Conclusions The subjective symptoms combined with conjunctival hyperemia can be more accurate assessment of ocular surface inflammation in the perioperative period of pterygium surgery,which can be used as an evaluation index to assess the effectiveness of anti-inflammatory treatment.