Abstract Fibroblast growth factor 2 (FGF2) is frequently induced during ischemic retinal injury and has traditionally been considered a pro-angiogenic factor based largely on studies using exogenous FGF2 administration. However, its endogenous cellular origin and physiological role remain incompletely understood. Here, we used single-cell transcriptomic analysis combined with spatial validation and rod photoreceptor-specific genetic approaches to define the endogenous role of FGF2 during oxygen-induced retinopathy (OIR). We identify rod photoreceptors as a major cellular source of ischemia-induced FGF2. Notably, Fgf2 expression remained elevated during the regression of pathological neovascularization, revealing a temporal dissociation between neuronal stress responses and vascular remodeling. Single-cell analysis further showed that Fgf2 induction occurred within a coordinated photoreceptor stress-response program involving endothelin 2 ( Edn2 ) and B-cell lymphoma 3 ( Bcl3 ). This transcriptional signature was independently reproduced in the N-methyl-N-nitrosourea (MNU)-induced photoreceptor degeneration model. Rod-specific deletion of Fgf2 markedly increased photoreceptor apoptosis, indicating that endogenous FGF2 contributes to photoreceptor survival under ischemic stress. In contrast, neither genetic depletion nor overexpression of FGF2 altered pathological neovascularization or vaso-obliteration. Bidirectional manipulation of FGF2 further modulated the expression of representative stress-associated genes Edn2 and Bcl3, supporting FGF2 involvement in this injury-response program. Finally, receptor expression analysis revealed relatively limited endothelial expression of Fgfr1-Fgfr4 compared with VEGF receptors, suggesting a cellular basis for the distinct effects of endogenous FGF2 and VEGF signaling. Together, these findings identify endogenous retinal FGF2 as a photoreceptor-derived survival factor that is induced during stress but is insufficient to drive pathological angiogenesis. These results support a model in which neuronal adaptation and vascular remodeling represent partially distinct responses during ischemic retinal injury.

This study aims to investigate the efficiency and the underlying mechanism of myeloid-derived suppressor cells (MDSCs) in corneal alkali burns (CAB). In the study, CD11b+ Gr-1+ cells from C57BL/6J mice bone marrow were cultured and induced. Cell activity and immunoregulatory function were assessed by flow cytometry in vitro. The optimal strategy of MDSCs therapy was assessed by slit-lamp microscopy, and flow cytometry in vivo. The therapeutic effects of MDSCs and the critical signaling pathway were investigated by hematoxylin-eosin (HE) staining, slit-lamp microscopy, flow cytometry, and immunofluorescence. The expression level of the NLRP3 inflammasome pathway was examined. The crucial biochemical parameters of MDSCs were examined by RNA-seq and qPCR to screen out the key regulators. The mechanism of MDSCs’ therapeutic effects was explored using MDSCs with IL-10 knockout/rescue by slit-lamp microscopy, HE staining, and qPCR evaluation. The cell frequencies of macrophages and neutrophils in the cornea were examined by flow cytometry in vivo. The results demonstrated that the induced MDSCs meet the standard of phenotypic and functional characteristics. The treatment of 5 × 105 MDSCs conjunctival injection on alternate days significantly ameliorated the disease development, downregulated the NLRP3 inflammasome pathway, and decreased the cell frequencies of macrophages and neutrophils in vivo significantly. IL-10 was screened out to be the critical factor for MDSCs therapy. The therapeutic effects of MDSCs were impaired largely by IL-10 knock-out and saved by the IL-10 supplement. In conclusion, MDSCs therapy is a promising therapeutic solution for CAB. MDSCs fulfilled immunoregulatory roles for CAB by IL-10-dependent anti-inflammatory properties.

Abstract Fungal pathogens are common causes of superficial clinical infection. Their increasing drug resistance gradually makes existing antifungal drugs ineffective. Heat stable antifungal factor (HSAF) is a novel antifungal natural product with a unique structure. However, the application of HSAF has been hampered by very low yield in the current microbial producers and from extremely poor solubility in water and common solvents. In this study, we developed an effective mode of treatment applying HSAF to superficial fungal infections. The marine‐derived Lysobacter enzymogenes YC36 contains the HSAF biosynthetic gene cluster, which we activated by the interspecific signaling molecule indole. An efficient extraction strategy was used to significantly improve the purity to 95.3%. Scanning electron microscopy images revealed that the Type I collagen‐based HSAF (Col‐HSAF) has a transparent appearance and good physical properties, and the in vitro sustained‐release effect of HSAF was maintained for more than 2 weeks. The effective therapeutic concentration of Col‐HSAF against superficial fungal infection was explored, and Col‐HSAF showed good biocompatibility, lower clinical scores, mild histological changes, and antifungal capabilities in animals with Aspergillus fumigatus keratitis and cutaneous candidiasis. In conclusion, Col‐HSAF is an antifungal reagent with significant clinical value in the treatment of superficial fungal infections.

Purpose: The purpose of this study was to investigate the characteristics and risk factors of intraocular lens (IOL) tilt and decentration of phacoemulsification after pars plana vitrectomy (PPV) using swept-source optical coherence tomography (SS-OCT). Methods: One hundred four eyes with prior PPV and 104 eyes without PPV undergoing uneventful cataract surgery were enrolled in this study. IOL tilt and decentration were measured by SS-OCT (CASIA2) 3 months postoperatively. Results: The mean IOL tilt and decentration were greater in the PPV group (5.36 ± 2.50 degrees and 0.27 ± 0.17 mm, respectively) than in the non-PPV group (4.54 ± 1.46 degrees, P = 0.005; 0.19 ± 0.12 mm, P < 0.001, respectively). Multiple logistic regression showed that silicone oil (SO) tamponade (odds ratio [OR] = 5.659, P = 0.021) and hydrophilic IOL (OR = 5.309, P = 0.022) were associated with IOL tilt over 7 degrees, and diabetes mellitus (DM; OR = 5.544, P = 0.033) was associated with IOL decentration over 0.4 mm. Duration of SO tamponade was positively correlated with IOL tilt (P = 0.014) and decentration (P < 0.001). The internal total higher-order aberration, coma, trefoil, and secondary astigmatism in the PPV group were higher than in the non-PPV group, and positively correlated with IOL tilt (P < 0.05). Conclusions: Patients with prior vitrectomy had greater IOL tilt and decentration than the non-PPV group. Longer duration of SO tamponade, hydrophilic IOL, as well as DM were the risk factors of greater IOL tilt and decentration in patients with prior PPV. Translational Relevance: Optically sophisticated designed IOLs should be used cautiously in vitrectomized eyes.

Background: To explore the performance of quantitative morphological and functional analysis in meibography images by an automatic meibomian glands (MGs) analyser in diagnosis and grading Meibomian Gland Dysfunction (MGD). Methods: A cross-sectional study collected 256 subjects with symptoms related to dry eye and 56 healthy volunteers who underwent complete ocular surface examination was conducted between January 1, 2019, and December 31, 2020. The 256 symptomatic subjects were classified into MGD group (n = 195) and symptomatic non-MGD group (n = 61). An automatic MGs analyser was used to obtained multi-parametric measurements in meibography images including the MGs area ratio (GA), MGs diameter deformation index (DI), MGs tortuosity index (TI), and MGs signal index (SI). Adjusted odds ratios (ORs) of the multi-parametric measurements of MGs for MGD, and the area under the receiver operating characteristic (AUC-ROC) curves of multi parametric measurements for MGD diagnosing and grading were conducted. Findings: When consider age, sex, ocular surface condition together, the estimated ORs for DI was 1.62 (95% CI, 1.29-2.56), low-level SI was 24.34 (95% CI, 2.73-217.3), TI was 0.76(95% CI, 0.54-0.90), and GA was 0.86 (95% CI, 0.74-0.92) for MGD. The combination of DI-TI-GA-SI showed an AUC = 0.82 (P < 0.001) for discriminating MGD from symptomatic subjects. The DI had a higher AUC in identifying early-stage MGD (grade 1-2), while TI and GA had higher AUCs in moderate and advanced stages (grade 3-5). Merging DI-TI-GA showed the highest AUCs in distinguish MGD severities. Interpretation: The MGs area ratio, diameter deformation, tortuosity and signal intensity could be considered promising biomarkers for MGD diagnosis and objective grading. Funding: This work was supported by the Key-Area Research and Development Program of Guangdong Province (No. 2019B010152001), the National Natural Science Foundation of China under Grant (81901788) and Guangzhou Science and Technology Program (202002030412). (c) 2021 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)

This study aimed to investigate the protective effect of melatonin on the corneal epithelium in dry eye disease(DED) and explore its underlying mechanism. Human corneal epithelial(HCE) cells was exposure to t-butylhydroperoxide(tBH), C57BL/6 mice were injected of subcutaneous scopolamine to imitate DED. Melatonin was used both in vivo and in vitro. Cell viability was detected by Cell Counting Kit-8 assay and Lactate Dehydrogenase Leakage. The change of cellular reactive oxygen species (ROS) levels, mitochondrial membrane potential (MMP), and apoptosis was analyzed by flow cytometry. Western blot assays and immunofluorescence were carried out to measure protein changes. mRNA expression was investigated by RNA sequencing (RNA-Seq) and quantitative real-time PCR. The change of autophagic flux were observed through mCherry-GFP-LC3 transfection and electron microscopy(TEM). Clinical parameters of corneal epithelium defects, conjunctival goblet cells, tear volume, and level of ocular surface inflammation was recorded. Melatonin was able to reduce excessive ROS production and maintain mitochondrial function. TEM assay found melatonin rescued impaired autophagic flux under tBH. Moreover, melatonin significantly preserved cell viability, abolished LDH release, and decreased apoptosis. RNA-Seq indicated that melatonin greatly activating hemeoxygenase-1 (HO-1) expression. Interestingly, HO-1 ablation largely attenuated its protective effects. Besides, in dry eye mouse model, intraperitoneal injection of melatonin showed greatly improved clinical parameters, inhibited activated NLRP3 inflammation cascade, and increased density of goblet cells and tear volume. Thus, melatonin protects corneal epithelial cells from oxidative damage, maintain normal level of autophagy, and reduce inflammation via trigging HO-1 expression in DED.

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.

Immune modulation plays a critical role in the pathogenesis of fungal keratitis (FK). However, the immune cell-mediated processes linking the innate immune response to the adaptive immune response are incompletely elucidated. IL-6 plays crucial roles in infectious and inflammatory processes in the cornea, regulating not only mononuclear macrophage differentiation but also lymphocyte activation, and IL-6 might be a useful target for immune intervention in FK. The frequencies of macrophages and T cells increased upon infection and were correlated with the severity of ocular pathogenesis. Additionally, protein profiling revealed that the expression of IL-6 and associated cytokines/chemokines was upregulated. Furthermore, anti-IL-6 intervention suppressed disease progression by reducing macrophage infiltration in the cornea and Th1, Th17, and Treg cell infiltration in draining lymph nodes (DLN) in an animal model of FK. Tocilizumab (TCZ), an antibody specific for IL-6, reduced the signal transducer and activator of transcription 3 (STAT3) activation in vivo and in vitro. In summary, fungal infection promoted macrophage and T cell activation via IL-6-mediated transcellular signaling to regulate immune cell migration and cytokine production, further demonstrating the role of IL-6 and providing a potential clinical therapeutic target in FK.

Accumulated oxidative damage may lead to irreversible retinal pigmented epithelium (RPE) cell death, which is considered to be the primary cause of dry age-related macular degeneration (AMD), leading to blindness in the elderly. However, an effective therapy for this disease is lacking. Here, we described a robust high-content screening procedure with a library of 814 protective compounds and found that D609 strongly protected RPE cells from sodium iodate (SI)-induced oxidative cell death and prolonged their healthy survival. D609 effectively attenuated excessive reactive oxygen species (ROS) and prevented severe mitochondrial loss due to oxidative stress in the RPE cells. Surprisingly, the potent antioxidative effects of D609 were not achieved through its own reducibility but were primarily dependent on its ability to increase the expression of metallothionein. The injection of this small water-soluble molecule also showed an explicit protective effect of the RPE layer in an SI-induced AMD mouse model. These findings suggested that D609 could serve as a novel antioxidative protector of RPE cells both in vitro and in vivo and unveiled a novel antioxidative mechanism of D609, which may ultimately have clinical applications for the treatment of AMD.

Purpose. To compare the prevalence of and factors associated with Demodex brevis and Demodex folliculorum in patients with cylindrical dandruff (CD group) and healthy controls. Methods. Eyelashes were taken from 1680 patients with CD and 1700 healthy controls in China from March 2015 to May 2017. All patients underwent a complete eye examination, and Demodex spp. were counted. The prevalence was analyzed according to age, gender, and clinical features. Results. Mean patient age was 42.93 ± 16.52 (3–88) and 39.4 ± 13.6 (7–81) years old in the CD and healthy control groups, respectively. In the CD and healthy groups, the positive rate for Demodex folliculorum was 27.92% and 8.47%, respectively, while that for Demodex brevis was 31.67% and 6.65%, respectively. In the CD group, the prevalence of Demodex brevis was higher than that of Demodex folliculorum, no matter in the females (33.65% versus 29.01%) or the males (28.54% versus 23.88%) in the CD group. Moreover, the numbers of Demodex folliculorum and Demodex brevis were significantly and positively correlated with age, in both children and old patients (both P<0.001), as well as with the severity of eyelid congestion (all P<0.05). Conclusions. In a large sample population, the prevalence of Demodex brevis and Demodex folliculorum was higher in the CD group than in healthy volunteers. In addition, the severity of eyelid congestion might be exacerbated by the number of Demodex spp., which may therefore provide a good clinical reference and objective guide.

Purpose:To assess how DNA damage-inducible transcript 4 (DDIT4) and autophagic flux are altered in dry eye disease and reveal the underlying mechanisms.Methods:C57BL/6 mice were exposed to desiccating stress (subcutaneous scopolamine [0.5 mg/0.2 mL] 3 times a day, humidity < 30%) for 7 days. Primary human corneal epithelial cells and cells from a human corneal epithelial cell line were cultured under hyperosmolarity. Western blot assays and immunofluorescence staining were used to measure changes in protein expression. mRNA expression was analyzed by RT-PCR and quantitative real-time PCR. Autophagosomes were observed through electron microscopy. Cellular reactive oxygen species and mitochondrial function were detected with 2',7'-dichlorodihydrofluorescein diacetate and mitochondrial membrane potential assays. Cell Counting Kit-8 and lactate dehydrogenase assays were used to measure cell death. Apoptosis was analyzed by Annexin V-PI flow cytometry.Results:Increased expression of microtubule-associated protein 1 light chain 3 (LC3-II), sequestosome 1 (SQSTM1), and DDIT4 were observed in corneal epithelial cells in in vitro and mice models of dry eye. The electron microscopy revealed large autophagic vacuoles with poorly degraded materials in human corneal epithelial cells under hyperosmolarity. In addition, we found that DDIT4 knockdown significantly suppressed the expression of LC3-II and SQSTM1 by disrupting reactive oxygen species release and restoring mitochondrial function under hyperosmolarity. Moreover, the ablation of DDIT4 effectively preserved cell viability and inhibited apoptosis.Conclusions:Excessive reactive oxygen species release through DDIT4 induction can lead to impaired autophagy and decreased cell viability in dry eye disease.

Background A worldwide lack of donor corneas demands the bioengineered corneas be developed as an alternative. The primary objective of the current study was to evaluate the efficacy of acellular porcine corneal stroma (APCS) transplantation in various types of infectious keratitis and identify risk factors that may increase APCS graft failure. Methods In this prospective interventional study, 39 patients with progressive infectious keratitis underwent therapeutic lamellar keratoplasty using APCS and were followed up for 12 months. Data collected for analysis included preoperative characteristics, visual acuity, graft survival and complications. Graft survival was evaluated by the Kaplan–Meier method and compared with the log-rank test. Results The percentage of eyes that had a visual acuity of 20/40 or better increased from 10.3% preoperatively to 51.2% at 12 months postoperatively. Twelve patients (30.8%) experienced graft failure within the follow-up period. The primary reasons given for graft failure was noninfectious graft melting (n = 5), and the other causes included recurrence of primary infection (n = 4) and extensive graft neovascularization (n = 3). No graft rejection was observed during the follow-up period. A higher relative risk (RR) of graft failure was associated with herpetic keratitis (RR = 8.0, P = 0.046) and graft size larger than 8 mm (RR = 6.5, P < 0.001). Conclusions APCS transplantation is an alternative treatment option for eyes with medically unresponsive infectious keratitis. Despite the efficacy of therapeutic lamellar keratoplasty with APCS, to achieve a good prognosis, restriction of surgical indications, careful selection of patients and postoperative management must be emphasized. Trial registration Prospective Study of Deep Anterior Lamellar Keratoplasty Using Acellular Porcine Cornea, NCT03105466. Registered 31 August 2016, ClinicalTrails.gov

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.

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.

This study was conducted to evaluate conjunctival blood flow velocities and microvascular network density in patients with dry eye disease (DED). Twenty-five patients with DED and 25 healthy controls were recruited. The microvasculature and microcirculation of the temporal bulbar conjunctiva of the right eyes were assessed using a functional slit-lamp biomicroscope. Vascular variables included blood flow velocity (BFV), blood flow rate (BFR), microvascular network density and vessel diameter. A fractal analysis was performed using the box counting method to measure the fractal dimension (Dbox) representing the vessel density. The bulbar BFV was 0.59 ± 0.09 mm/s in the DED group and 0.47 ± 0.12 in the control group (P < 0.001). BFR was 169.5 ± 1.8 in the DED group compared to the control group (107.2 ± 49.6) (P < 0.001). Dbox was higher in DED patients (1.65 ± 0.04) than controls (1.60 ± 0.07, P < 0.05). Moreover, the vessel diameter was larger in the DED group (21.8 ± 1.8 μm) compared with controls (17.9 ± 2.2 μm, P < 0.001). Dbox was positively related with ocular surface disease index (OSDI) in patients with DED (r = 0.54, P = 0.008). Microvascular alterations were found in the bulbar conjunctiva of DED patients, including increased blood flow velocity, higher vessel density and larger vessel diameter.

PURPOSE:To establish a new scoring system for limbal dermoid, in order to unify the diagnostic criteria and assess the prognosis.METHODS:A retrospective study was conducted on 261 patients with limbal dermoid. The basic information, clinical features, and pathology of dermoids were recorded, and the prognosis at 1 year after keratoplasty was assessed at follow-up. A new visual scoring system was created for the area of corneal involvement, the area of conjunctival involvement, and the surface shape.RESULTS:There were 154 females and 107 males with mean age of 4 ± 3 years at surgery. After scoring, 59% (136) of patients were classified as grade I, 26% (60) as grade II, and 14% (33) as grade III. The pathological results were 124 dermoid cases, 76 lipodermoid, 5 complex choristoma, and 10 epibulbar osseous choristoma. Moreover, patients with lower clinical scores presented a better prognosis; the mean logarithm of the minimum angle of resolution (logMAR) best-corrected visual acuity in grade I patients was 0.38 ± 0.05, which was better than the grade II value of 0.61 ± 0.09 (P < 0.05) and the grade III value of 0.94 ± 0.11 (P < 0.001).CONCLUSIONS:New grading systems for limbal dermoid were useful for clinical diagnosis and may have prognostic value in predicting visual acuity. A lower-grade dermoid exhibited better vision postoperatively.

Lipopolysaccharide (LPS)-induced keratitis is a progressive infectious ocular disease in which innate inflammatory responses often cause clinical tissue damage and vision loss. The present study aimed to investigate the effects of tacrolimus, an effective immunomodulator, on LPS-induced innate immune responses. The effects of tacrolimus on the apoptotic rate and viability of human corneal epithelial cells (HCECs), polymorphonuclear neutrophils (PMNs) and monocytes (THP-1 cells) were examined using flow cytometry and MTT assays. Subsequently, the role of tacrolimus on LPS-induced inflammation in HCECs, PMNs and THP-1 cells was evaluated by detecting the expression levels of pro-inflammatory cytokines, including interleukin (IL)-1β, IL-6 and matrix metallopeptidase 9; anti-inflammatory cytokines, including IL-10 and transforming growth factor-β; and proangiogenic factors, including vascular endothelial growth factor and tumor necrosis factor-α using quantitative polymerase chain reaction. The results demonstrated that tacrolimus had good biocompatibility with HCECs, while promoting apoptosis and decreasing the viability of PMNs and THP-1 cells. Furthermore, tacrolimus effectively reduced the expression levels of pro-inflammatory cytokines and increased anti-inflammatory cytokines in LPS-induced keratitis in vitro. Notably, tacrolimus decreased the levels of proangiogenic factors, which are highly increased following LPS stimulation. Conclusively, tacrolimus appears to be a safe and effective treatment to suppress neutrophil and monocyte activity, modulate the balance of pro-/anti-inflammatory cytokines, and reduce the inflammatory response and angiogenic activity in LPS-induced bacterial keratitis.