Purpose:This prospective cohort study aimed to assess the influence of tear film stability on corneal refractive power measurement and surgical planning in cataract patients. Methods:Participants were divided into tear film instability (tear film stability level 2) and control (level 0-1) groups based on Keratograph 5M results. Using IOL Master 700, two consecutive measurements were obtained with a 10-min interval. Parameters including standard keratometry (Kf, Ks and K), keratometric corneal astigmatism (KCA), total keratometry (TKf, TKs and TK) and total corneal astigmatism (TCA) were recorded. IOL power was calculated using SRK-T, SRK-T TK, Haigis, Haigis TK, Barrett Universal II, and Barrett Universal II TK formulas. Results:The results showed significantly higher differences between two measurements in Kf, K, KCA, TKf, TK, and TCA, as well as the vector variability of corneal astigmatism in the tear film instability group (all P ​< ​0.05). Of all formulas, only the SRK-T formula displayed significantly higher variability in IOL power calculations in the tear film instability group compared to the control group (P ​< ​0.05). Conclusions:This study highlights that tear film instability can lead to deviations in corneal refractive power and astigmatism measurements, contributing to increased prediction errors in IOL power calculation, particularly with the SRK-T formula.

Ethnopharmacological relevanceQiju Dihuang Pill (QDP) is a traditional Chinese medicine prescription for the treatment of eye diseases. Novel literature reports that copper-induced cell death, called as cuproptosis, is a copper-dependent and differs distinctly from other types of cell death.Aim of the studyThe present study aims to investigate whether QDP could protect lens epithelial cells via alleviating copper-induced death in diabetic cataract.Materials and methodsThe different concentration of QDP medicated serum was administrated on high glucose (HG)-induced human lens epithelial cells (HLECs). The copper concentration was tested using Elabscience Copper Assay kit. The proliferation was detected using CCK-8 and EdU assays. The molecular binding was identified using RIP-PCR and luciferase reporter assay.ResultsResults indicated that HG culture condition triggered the copper concentration and repressed the proliferation of HLECs. Then, the elesclomol-Cu (Es-Cu) administration up-regulated the copper concentration and inhibited the proliferation, and cuproptosis inhibitor tetrathiomolybdate (TTM) could specifically reverse the consequence. QDP treatment reduced the copper concentration and cuproptosis-related genes (SLC31A1, FDX1). MeRIP-Seq and RIP-PCR confirmed that QDP reduced the stability of SLC31A1 mRNA through m6A modified site, and copper actually synergized the molecular binding efficiency. Rescue assay verified the role of QDP and SLC31A1 on HLECs’ cuproptosis characteristic.ConclusionThis research identified the protective role of QDP on HG-induced HLECs in DC through decreasing m6A/SLC31A1-mediated cuproptosis in DC. This finding provides novel insights into mechanisms for QDP and sheds light on the multifaceted role of traditional prescription on DC.

A retrospective cohort study was conducted to observe the correction effect of Toric intraocular lens (IOL) implantation in cataract eyes with specific types of irregular corneal astigmatism. Thirty-four eyes with either the "asymmetric bow-tie" pattern (Type I) or the "angled bow-tie" pattern (Type II) were included. Corneal topography was assessed using Pentacam HR, and changes in preoperative corneal astigmatism, visual acuity, manifest refraction, and objective visual quality were measured and compared. The average uncorrected distance visual acuity improved significantly from 0.86 ± 0.40 logMAR to 0.22 ± 0.15 logMAR (P < 0.001). Preoperative corneal astigmatism of 2.05 ± 0.90 D was corrected to a postoperative residual astigmatism of 0.78 ± 0.57 D (P < 0.001), with 32% of eyes within 0.50 D. The residual astigmatism prediction errors in Type I and Type II cases were (0.97 ± 0.68 D) and (0.66 ± 0.37 D), respectively (P = 0.100). The mean spherical equivalent prediction error in Type II cases (0.07 ± 0.36 D) was significantly smaller than that in Type I cases (− 0.29 ± 0.52 D) (P = 0.030). This study concludes that Toric IOL implantation effectively corrects specific types of irregular corneal astigmatism in cataract surgery. Eyes with the "angled bow-tie" pattern show higher accuracy in refractive predictions compared to eyes with the "asymmetric bow-tie" pattern.

Intraocular irrigating solution is extensively applied in cataract surgery. This paper explored the difference and relationship between optical coherence tomography (OCT) and optical quality analysis system (OQAS) parameters induced by compound electrolyte intraocular irrigating solution (CEIIS) or Ringer lactate (RL) solution during uncomplicated cataract surgery. Totally 200 senior cataract patients were randomly divided into the CEIIS and RL groups (N = 100 patients/group). The anterior chamber was irrigated by CEIIS or RL during phacoemulsification. Patients were subdivided into diabetes mellitus (DM)+ and DM− groups. The central macular thickness (CMT), hyper reflective foci (HF), modulation transfer function cutoff frequency (MTF cutoff), Strehl ratio (SR), objective scatter index (OSI), and OQAS values (OVs) at 100

Purpose: To evaluate and compare the accuracy of iTrace and CASIA2 in measuring the postoperative orientation of toric intraocular lens (IOL) without mydriasis. Setting: Tianjin Medical University Eye Hospital, Tianjin, China. Design: Prospective cohort study. Methods: Patients with SN6AT toric IOLs implanted after cataract surgery were enrolled. 1 month after surgery, the toric IOL orientation were measured by iTrace and CASIA2 in non-mydriatic, semi-dark conditions. Then, the toric axis was directly reviewed using the slit-lamp under full mydriasis. Axis measurement differences between each of the 2 devices and the slit-lamp, described as their relative differences (RDs), were calculated and compared. The percentage of RDs within 5 degrees, within 10 degrees and greater than 30 degrees were analyzed. Results: 77 eyes of 70 patients were included. Generally, the mean toric axis measurement RDs of CASIA2 and iTrace were 9.24 ± 10.53 degrees and 13.89 ± 15.47 degrees respectively (P = .04). For CASIA2 (72 eyes), 54.17% (39), 72.22% (52), and 4.17% (3) of eyes had RDs within 5 degrees, within 10 degrees and greater than 30 degrees, compared with 40.00% (28), 61.43% (43) and 12.86% (9) for iTrace (70 eyes). The 95% limits of agreements of CASIA2 with slit-lamp was narrower than that of iTrace with slit-lamp. The median RD of CASIA2 was significantly smaller in eyes with pupil ≥4 mm under dark condition compared with eyes with pupil <4 mm (P = .03). Conclusions: CASIA2 demonstrates greater precision in measuring toric IOL orientation under non-mydriatic conditions compared with iTrace. Moreover, the accuracy of CASIA2 is enhanced in cases of pupil >4 mm.

Purpose: To investigate the influence of posterior corneal astigmatism on the prediction accuracy of toric multifocal intraocular lens (IOL) calculation. Methods: The keratometric astigmatism measured by Lenstar LS 900 (KCA(L)), keratometric astigmatism (KCA(P)) and total corneal astigmatism (TCA) measured by Scheimpflug camera (Pentacam HR) were documented and analyzed accordingly. Three deduction models using different parameters were compared. Model 1: KCA(L) + keratometric corneal surgically induced astigmatism (KCSIA, 0.30 D @ 50 degrees); Model 2: KCA(P) + KCSIA); Model 3: TCA + total CSIA (TCSIA, 0.23 D @ 50 degrees). The prediction errors of each model as the difference vector between the actual and the intended residual astigmatism were compared. Results: Seventy-six eyes implanted with toric multifocal IOLs were included in this study. The vector differences of the actual KCSIA and TCSIA were statistically significant in the total sample and against-the-rule (ATR) subgroup (both P < 0.05). Model 1 deduced the smallest mean values of prediction error, while that of Model 3 were smaller than that of Model 2, both in the total sample and the ATR subgroups (all P < 0.05). Meanwhile, in the total sample and ATR subgroups, the centroid vector magnitudes of Model 3 were smaller than that of Model 1 (0.31 +/- 0.76 D and 0.39 +/- 0.76 D). Conclusions: The calculation of toric multifocal IOL should be individualized especially in the ATR eyes for the impact of PCA on the estimation of the preoperative corneal astigmatism and the CSIA.

Fungal hyphae deeply invade the cornea in fungal keratitis. The corneal stroma hinders the infiltration of antifungal drugs and reduces their bioavailability. Here, this work reports a peptide conjugate nano-assembly that permeates the stroma and kills the pathogen without irritating the ocular cornea. The hydrophilic surface of the nano-assembly ensures deep permeation into the stroma. When encountering a fungal hyphal cell, the nano-assembly disassembles and exposes the α-helical peptide to destroy the fungal membrane, thus inactivating the pathogen. In a rabbit model of fungal keratitis, the nano-assembly exhibits a better therapeutic effect than commercially available natamycin ophthalmic suspension. Peptide conjugates with a nano-assembled structure and assembly-disassembly behavior could serve as the foundation of a new therapy for fungal keratitis.

Objective::To compare the difference and agreement of corneal total higher-order aberration (tHOA) and corneal spherical aberration (SA) measured by the new generation of refractive analyzer (OPD-ScanⅢ), swept-source anterior segment optical coherence tomography (SS-AS-OCT) device (CASIA2), ray-tracing aberrometer (iTrace) and 3D anterior segment diagnostic analyzer (Pentacam) in cataract patients.Methods::In this case series study, a total of 123 patients (123 eyes) diagnosed with cataract were selected before phacoemulsification in the Tianjin Medical University Eye Hospital during May to August 2021. All the patients were examined for corneal parameters by iTrace, Pentacam, OPD-ScanⅢ and CASIA2. The root mean square (RMS) of corneal total higher-order aberration (tHOA) in the central 4 mm and corneal spherical aberration (SA) in the central 6 mm of the corneal apex position were recorded. The difference of measurement results between four devices were analyzed by Two-way analysis of variance or Friedman test. Correlation was evaluated using Person or Spearman correlation analysis and the agreement was evaluated using Bland-Altman plots.Results::There were significant differences between four devices for corneal tHOA ( Z=134.79, P<0.001). There was no significant difference between CASIA2 and Pentacam or OPD-ScanⅢ and iTrace for corneal tHOA. The corneal tHOA measured by Pentacam and CASIA2 were higher than that measured by OPD-ScanⅢ and iTrace ( P<0.001). There were significant differences between four devices for corneal SA ( F=114.72, P<0.001). The corneal SA was 0.291±0.079 μm, 0.445±0.147 μm, 0.310±0.092 μm and 0.277±0.131 μm respectively by iTrace, Pentacam, OPD-ScanⅢ and CASIA2. There was no statistically significant difference between OPD-ScanⅢ and iTrace or CASIA2 and iTrace. Pairwise comparison of the corneal SA in other groups showed significant difference ( P<0.05). The results of corneal tHOA and corneal SA measured by four devices were positively correlated ( r=0.27-0.69, P<0.001). Bland-Altman analysis showed that the 95% limits of agreement (95%LoA) of corneal tHOA and corneal SA measured by iTrace and OPD-ScanⅢ were -0.11-0.09 and -0.15-0.12, which showed clinically acceptable agreement. The results of corneal tHOA and corneal SA measured by iTrace and OPD-ScanⅢ were 4.07% to 5.69% outside 95%LoA. Other results had poor consistency with a wide 95%LoA. Conclusions::The results of corneal tHOA in the central 4 mm and corneal SA in the central 6 mm of the corneal apex position measured by OPD-ScanⅢ and iTrace showed no difference, good agreement and consistency. However, the other measurement results of the four devices were different.

Fungal keratitis (FK) is a refractory disease that poses a serious threat to vision, with common risk factors like eye trauma, contact lens wearing, topical corticosteroids and antibiotic abuse. Nowadays, topical and systemic anti-fungal drugs and ocular surgeries are still the main therapeutic modalities. However, the pathogenesis of FK, especially the immunologic mechanism within it, has not yet been deeply clarified. A better understanding of the pathogenesis of FK is imperative for more effective therapies and prognosis. Meanwhile, the immune protection strategies are also urgently required to manage FK. This review highlights recent advances in the immunologic mechanism in the pathogenesis of FK, in hope of providing valuable reference information for more effective anti-fungal treatment.

An important mechanism involved in dry eye (DE) is the association between tear hyperosmolarity and inflammation severity. Inflammation in DE might be mediated by the NLRP3 inflammasome, which activated by exposure to reactive oxygen species (ROS). A combination of carboxymethylcellulose (CMC) and α-melanocyte stimulating hormone (α-MSH) may influence DE through this mechanism, thus avoiding defects of signal drug. In this study, we assessed whether treatment comprising CMC combined with α-MSH could ameliorate ocular surface function; we found that it promoted tear secretion, reduced the density of fluorescein sodium staining, enhanced the number of conjunctival goblet cells, and reduced the number of corneal apoptotic cells. Investigation of the underlying mechanism suggested that the synergistic effect of combined treatment alleviated DE inflammation through reduction of ROS level and inhibition of the NLRP3 inflammasome in human corneal epithelial cells. These findings indicate that combined CMC + α-MSH treatment could ameliorate lesions and restore ocular surface function in patients with DE through reduction of ROS level and inhibition of NLRP3 signalling.

Rationale: Corneal transplantation is an effective treatment to corneal blindness. However, the immune rejection imperils corneal allograft survival. An interventional modality is urgently needed to inhibit immune rejection and promote allograft survival. In our previous study, subconjunctival injections of bone marrow-derived mesenchymal stem cells (BM-MSCs) into a rat model of corneal allograft rejection extended allograft survival for 2 d. In this study, we sought to generate IL-10-overexpressing BM-MSCs, aiming to boost the survival-promoting effects of BM-MSCs on corneal allografts and explore the molecular and cellular mechanisms underlying augmented protection. Methods: A population of IL-10-overexpressing BM-MSCs (designated as IL-10-BM-MSCs) were generated by lentivirus transduction and FACS purification. The self-renewal, multi-differentiation, and immunoinhibitory capabilities of IL-10-BM-MSCs were examined by conventional assays. The IL-10-BM-MSCs were subconjunctivally injected into the model of corneal allograft rejection, and the allografts were monitored on a daily basis. The expression profiling of long noncoding RNA (lncRNA) in the allografts was revealed by RNA sequencing and verified by quantitative real-time PCR. The infiltrating immune cell type predominantly upregulating the lncRNA expression was identified by RNAscope in situ hybridization. The function of the upregulated lncRNA was proved by loss- and gain-of-function experiments both in vivo and in vitro. Results: The IL-10-BM-MSCs possessed an enhanced immunoinhibitory capability and unabated self-renewal and multi-differentiation potentials as compared to plain BM-MSCs. The subconjunctivally injected IL-10-BM-MSCs reduced immune cell infiltration and doubled allograft survival time (20 d) as compared to IL-10 protein or plain BM-MSCs in the corneal allograft rejection model. Further, IL-10-BM-MSCs significantly upregulated lncRNA 003946 expression in CD68+ macrophages infiltrating corneal allografts. Silencing and overexpressing lncRNA 003946 in macrophage cultures abolished and mimicked the IL-10-BM-MSCs' suppressing effects on the macrophages' antigen presentation, respectively. In parallel, knocking down and overexpressing the lncRNA in vivo abrogated and simulated the survival-promoting effects of IL-10-BM-MSCs on corneal allografts, respectively. Conclusion: The remarkable protective effects of IL-10-BM-MSCs support further developing them into an effective interventional modality against corneal allograft rejection. IL-10-BM-MSCs promote corneal allograft survival mainly through upregulating a novel lncRNA expression in graft-infiltrating CD68+ macrophages. LncRNA, for the first time, is integrated into an IL-10-BM-MSC-driven immunomodulatory axis against the immune rejection to corneal allograft.

Objective:To evaluate the clinical efficacy and safety of modified chitosan eye drops on rabbit Candida albicans keratitis model.Methods:Ten healthy female New Zealand rabbits were used to establish the superficial Candida albicans keratitis model by the corneal surface lens method in the right eye.Slit lamp microscopy and corneal scraping and microscopic examination were performed to preliminarily determine whether the keratitis model has been successfully established, the rabbits were then randomly divided into a model group and a modified chitosan group by the random number table method.The successfully established rabbit models which were determined by fungal culture results were retained.Five normal rabbits receiving no intervention served as a normal control group.The experimental eyes in the normal control and modified chitosan groups were treated with modified chitosan eye drops, Six times a day for one week, and subsequently four times a day for one week.No treatment was administered to the model group.The changes of corneal lesions and ocular surfaces were examined by slit lamp microscopy every day.At 1, 7, 14, 21 and 28 days after modeling, the eye condition and corneal clinical scores was assessed by slit lamp microscopy.The corneal conditions in each group was observed for two weeks after drug withdrawal.Results:The corneal scraping and microscopic examination results of eight rabbits models showed that the fungal hyphae and spores were positive.The fungal culture results showed that the separated pathogen was Candida albicans.The success rate of modeling was 80%(8/10). The clinical scores in the model group at 7, 14 and 21 days after modeling were 14.50±0.58, 6.25±0.50 and 2.50±0.58, respectively, and were significantly higher than 7.25±1.26, 2.75±0.50 and 1.25±0.50 in the modified chitosan group (all at P<0.05). In the model group, corneal edema was significantly aggravated, and the central white ulcer area was enlarged within seven days after modeling.Between 7 and 28 days after modeling, the corneal ulcer was gradually healed, while the central corneal scar and neovascularization were remained.The average healing time was (24.5±2.6)days.In the modified chitosan group, the corneal infiltration was significantly alleviated within seven days after modeling, and the fungal hyphae and spores of corneal scraping were negative on the 14th day after modeling.The average healing time in the modified chitosan group was (13.5±1.3)days, which was significantly shorter than that in the the model group (P<0.01). No recurrence of keratitis was observed in the modified chitosan group after two weeks of drug withdrawal.The cure rate was 100%.In the normal control group, the conjunctival hyperemia, corneal edema, and lesions were not observed during topical administration.Conclusion:The treatment with modified chitosan eye drop is effective in a rabbit superficial Candida albicans keratitis model, and have no obvious toxic effects on ocular tissues.

Oxidative stress is widely known to be a major contributor in the pathogenesis of dry eye disease (DED). 4-Hydroxynonenal (4-HNE), a well-known byproduct frequently measured as an indicator of oxidative stress-induced lipid peroxidation, has been shown to be elevated in both human and murine corneal DED samples. This study aims to investigate if 4-HNE is responsible for the oxidative stress in human corneal epithelial cells (HCECs) and explores the underlying mechanism by which it confers its effects. SV40-immortalized HCECs were cultured in minimum essential media (MEM) with 1% penicillin/streptomycin and 10% fetal bovine serum. HCECs were exposed to media with or without 4-HNE and cell culture supernatants were collected at 4 and 24 h. Cellular reactive oxygen species (ROS) measurement was performed using a 2′,7′-dichlorofluorescein diacetate (DCFDA) assay kit according to the manufacturer’s instructions. Protein levels of antioxidant enzymes copper/zinc superoxide dismutase 1 (SOD1) and NAD(P)H quinone dehydrogenase 1 (NQO1) were analyzed by Western blot. NF-κB activation and expression of IL-6 and IL-8 were measured using an NF-κB p65 Total SimpleStep ELISA Kit and Proteome Profiler Human Cytokine Array Kit. Cell viability was evaluated by LDH cytotoxicity assay. Treatment with 4-HNE decreased cell viability of HCECs. Band intensities corresponding to levels of ROS production showed a significant increase in ROS generation after treatment with 4-HNE. 4-HNE decreased SOD1 levels and upregulated NQO1 expression in HCECs. A significant increase in activation of NF-κB and production of pro-inflammatory cytokines IL-6 and IL-8 was observed after treatment with 4-HNE. Exposure to N-acetylcysteine (NAC), an antioxidant and ROS scavenger, antagonized the oxidative effects of 4-HNE on HCECs. 4-HNE induces oxidative stress in corneal epithelial cells by increasing levels of ROS generation and modifying the expression of antioxidant enzyme levels, decreasing cell viability of HCECs in vitro. This study demonstrates a potential pathway by which 4-HNE functions to confer its detrimental effects and provides a new therapeutic target for the treatment of DED.

As a deacetylation product of chitin, chitosan is the only basic cationic polysaccharide. It has been widely used in the medical field, such as wound healing, drug carriers, and tissue engineerin, because its good biocompatibility, non-toxicity, low sensitivity and biodegradability. Based on the biological characteristics of chitosan and its derivatives, this article will review the application and research of chitosan in the following ophthalmic subspecialties: sustained release of drugs for ocular surface diseases (such as eyelid defects, dry eyes, keratitis, corneal wound healing, corneal tissue engineering); chitosan coating complex liposome which can delay cataract formation, sustained release of antimetabolite from chitosan nanoparticles modified intraocular lens to prevent posterior capsule opacification(PCO); anti-glaucoma drug sustained release for glaucoma treatment and anti-scarring drug sustained release post filter surgery; chitosan oligosaccharides’ effect in experimental autoimmune anterior uveitis; retinopathy (insulin sustained release for diabetic retinopathy, a kind of new scleral buckling material for retinal detachment, and chitosan oligosaccharides prevent retina from ischemia-reperfusion injury). However, most researches are in vitro or animal experiments, and there are no large sample size and rigorously designed long-term clinical trials. ( Int Rev Ophthalmol, 2020, 44: 105-111)

Objective To investigate the inhibitory effect of bromfenac sodium hydrate ophthalmic solution on corneal neovascularization (CNV) induced by alkali burn.Methods A total of 192 specific pathogen free (SPF) degree adult male Sprague-Dawley (SD) rats were used in this study.One hundred and seventy-two rats were chosen to establish CNV model with alkali burn in the right eyes.Following alkali burn,rats were randomly divided into CNV group,model control group,bromfenac sodium group and fluorometholone group,with 43 rats (43 eyes) in each group.Another 20 rats (40 eyes) served as normal control group.One day after modeling,the model control group,bromfenac sodium group and fluorometholone group received phosphate buffer saline (PBS),bromfenac sodium hydrate ophthalmic solution and 0.1％ fluorometholone eye drops,respectively.The state of cornea and anterior chamber and the growth of CNV of rats in each group were observed by slit-lamp microscope every day after modeling.At 1,3,7,14,21 and 28 days after modeling,the anterior segment photos of the experimental eyes were captured,and the percent of cornea areas covered by CNV was calculated.At 7,14 and 28 days after modeling,the eye tissue sections were stained with hematoxylin and eosin staining and immunohistochemistry staining to evaluate the expressions of CD45 and VEGF-A.Real-time quantitative PCR and enzyme-linked immunosorbent assay (ELISA)were used to detect the expression of COX-2 and VEGF mRNA and protein level.The use and care of the animals complied with the Statement of the Association for Research in Vision and Ophthalmology(ARVO).Results Each model group showed corneal edema and opacification 1 day after modeling.The corneal edema was aggravated 7 days after modeling.On the 14th day after modeling,the degree of corneal opacity and edema decreased gradually.On the 28th day after modeling,leucoma was observed in CNV group and model control group,and nebula was observed in bromfenac sodium group and fluorometholone group.At 7,14,21 and 28 days after modeling,the percentages of CNV areas in bromfenac sodium group and fluorometholone group were significantly lower than those in CNV group and model control group (all at P＜0.05).No significant difference was found in the percentage of CNV areas between bromfenac sodium group and fluorometholone group at various time points (all at P＞0.05).On the 7th day after modeling,the thinning of corneal epithelial layer,edema and arrangement disorder of stroma layer were observed,and the expression of VEGF-A was positive in all model groups;a small amount of CD45 positive inflammatory cell infiltrations were observed in CNV group and model control group.On the 14th and 28th day after modeling,CNV was seen in the center of cornea in CNV group and model control group;the epithelial keratosis and reduction of corneal edema were seen in each group,and no inflammatory cell infiltration was observed in each group.On the 7th day after modeling,the expressions of COX-2 and VEGF mRNA in CNV group and model control group were significantly higher than those in normal control group,bromfenac sodium group and fluorometholone group (all at P ＜ 0.05),the expressions of COX-2 and VEGF protein in bromfenac sodium group were significantly lower than those in CNV group (all at P＜0.05).The corneal peroration rate in model control group and bromfenac sodium group was 10％ (1 case in 10 rats).The corneal perforation rate in fluorometholone group was 30％ (3 cases in 10 rats).In each model group,10％ to 30％ rats had hyphema.Conclusions Bromfenac sodium hydrate ophthalmic solution can inhibit the formation and growth of CNV after alkali burn in rats.This effect may be mediated by regulating COX-2 expression,reducing inflammation and inhibiting VEGF production.

Objective The immune rejection mediated by CD4+ T cell and antigen presenting macrophages is the leading cause of corneal transplantation failure. Bone marrow-derived mesenchymal stem cells (BM-MSCs) possess robust immunomodulatory potentials, and have been shown by us and others to promote corneal allograft survival. However, the immunological mechanism underlying the protective effects of BM-MSCs remains unclear. Therefore, in the current study, this mechanism was investigated in a BM-MSC-treated rat model of corneal allograft rejection, in the hope to facilitate the search for novel interventional targets to corneal allograft rejection. Methods Lewis rats were subjected to corneal transplantation and then received subconjunctival injections of BM-MSCs (2×106 cells / 100 μl PBS) immediately and at day 3 post-transplantation. The control group received the injections of PBS with the same volume. The clinical parameters of the corneal allografts, including opacity, edema, and neovascularization, were regularly evaluated after transplantation. On day 10 post-transplantation, the corneal allografts were collected and subjected to flow cytometry and high-throughput RNA sequencing (RNA-seq). GO enrichment and KEGG pathways were analyzed. The quantitative realtime PCR (qPCR) and immunohistochemistry (IHC) were employed to validate the expression of the selected target genes at transcript and protein levels, respectively. Results BM-MSC subconjunctival administration prolonged the corneal allograft survival, with reduced opacity, alleviated edema, and diminished neovascularization. Flow cytometry showed reduced CD4+ T cells and CD68+ macrophages as well as boosted regulatory T cells (Tregs) in the BM-MSC-treated corneal allografts as compared with the PBS-treated counterparts. Moreover, the RNA-seq and qPCR results demonstrated that the transcript abundance of Cytotoxic T-Lymphocyte Associated Protein 4 (Ctla4), Protein Tyrosine Phosphatase, Receptor Type C (Ptprc), and C-X-C Motif Chemokine Ligand 9 (Cxcl9) genes were increased in the allografts of BM-MSC group compared with PBS group; whereas the expression of Heat Shock Protein Family A (Hsp70) Member 8 (Hspa8) gene was downregulated. The expression of these genes was confirmed by IHC at protein level. Conclusion Subconjunctival injections of BM-MSCs promoted corneal allograft survival, reduced CD4+ and CD68+ cell infiltration, and enriched Treg population in the allografts. The BM-MSC-induced upregulation of Ctla4, Ptprc, Cxcl9 genes and downregulation of Hspa8 gene might contribute to the protective effects of BM-MSCs and subserve the potential interventional targets to corneal allograft rejection.

PURPOSE:To observe ocular surface changes in Type II diabetic patients with different disease durations and to understand the correlations between clinical parameters and diabetic durations.METHODS:In this cross-sectional, prospective study, 51 healthy controls and 91 patients with Type II diabetes were enrolled. The diabetics were divided into 3 subgroups according to the disease duration, including duration <10 y group, 10 to 20 y group, and ≥21 y group. All subjects underwent clinical ocular examinations, including lipid layer thickness (LLT), blinking rate, tear meniscus height (TMH), noninvasive tear film break-up time (NI-BUT), meibography, superficial punctate keratopathy (SPK) scoring, corneal sensitivity, and Schirmer I test. They were also evaluated using the standard patient evaluation of eye dryness (SPEED) questionnaire.RESULTS:SPEED score, meiboscore, SPK score, LLT, Schirmer I test, and corneal sensitivity differed significantly between the diabetic and healthy control groups. Further, SPEED score, Schirmer I test, corneal sensitivity, meiboscore, and blink rate significantly differed among the 3 diabetic subgroups and the control group. In diabetics, the SPEED score correlated with the SPK score, blink rate, TMH, and LLT; NI-BUT with TMH, LLT, and blink rate; TMH with the SPK score; Schirmer I test with the SPK score; and corneal sensitivity with the meiboscore. More importantly, the Schirmer I test, corneal sensitivity, and SPEED score negatively correlated with diabetic duration.CONCLUSION:Diabetic duration is an important factor that affects functions of the lacrimal functional unit in patients with Type II diabetes. The trends of changes in the ocular parameters vary along the course of diabetes.

Video display terminal (VDT) related dry eye,which can impair eye health significantly and has become one of the main clinical subdiseases in Ophthalmology.The pathogenesis of VDT related dry eye is complicated,involving mechanical and non-mechanical mechanisms.The mechanical mechanism is mainly related with the blink movement.It includes the reduction of complete blink,increase of incomplete blink,and increase of ocular surface exposed area.Non-mechanical mechanisms include ocular surface inflammation,decrease of mucin concentration and lacrimal gland function.The specific examination of VDT related dry eye haven't been elucidated,and the clinical recommendation is to use the questionnaire combined with symptoms and routine dry eye examinations to diagnose this disease.