Publications by authors named "Weiyun Shi"

119 Publications

A proteomic approach towards understanding the pathogenesis of Mooren's ulcer.

Exp Eye Res 2021 Feb 27;205:108509. Epub 2021 Feb 27.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China; Eye Hospital of Shandong First Medical University, Jinan, Shandong Province, China. Electronic address:

Mooren's ulcer (MU) is a refractory autoimmune corneal ulcer with a high recurrence rate. So far, its molecular profiles and pathomechanisms remain largely unknown. Therefore, we aim to characterize the protein profiles of MU specimens by data-independent-acquisition (DIA) mass spectrometry (MS), and to define the functions of differentially-expressed proteins (DEPs). Through LC-MS/MS, 550 DEPs were identified between MU biopsies and age-matched controls (Ctrl). KEGG analysis revealed that the significantly enriched pathways of the up-regulated proteins mainly covered lysosomes, antigen processing and presentation, and phagosomes. We subsequently validated the expressions of the selected candidates using parallel-reaction-monitoring (PRM)-based MS and immunohistochemistry (IHC), including cathepsins, TIMP3, MMP-10, MYOC, PIGR, CD74, CAT, SOD2, and SOD3. Moreover, immunoglobulin (Ig) components and B lymphocytes associated proteins MZB1, HSPA5, and LAP3 in MU were significantly increased and validated by PRM-based MS and IHC. The remarkable enrichment of neutrophil extracellular traps (NETs) components in MU samples was also identified and determined. The up-regulated Ig components and NETs components suggested that B lymphocytes and neutrophils participated in the immunopathology of MU. Importantly, we also identified and validated much more expression of peptidyl arginine deiminase 4 (PADI4) in MU samples. The double-immunofluorescence staining showed the co-localization of citrulline residues with MPO, NE, and IgG in MU samples. These results indicated the presences of PADI4-mediated citrullination modification and anti-citrullinated protein antibodies (ACPAs) in MU samples. Our findings, for the first time, provide a global proteomic signature of MU, which may open a new avenue towards disease pathology and therapeutics.
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http://dx.doi.org/10.1016/j.exer.2021.108509DOI Listing
February 2021

Multiple roles of FGF10 in the regulation of corneal endothelial wound healing.

Exp Eye Res 2021 Feb 20;205:108517. Epub 2021 Feb 20.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China; Shandong Eye Hospital, Shandong Eye Institute, Shandong First Medical University &Shandong Academy of Medical Sciences, China. Electronic address:

Corneal endothelial dysfunction usually induces corneal haze and oedema, which seriously affect visual function. The main therapeutic strategy for this condition is corneal transplantation, but the use of this strategy is limited by the shortage of healthy donor corneas. Compared with corneal transplantation, drug intervention is less invasive and more accessible; thus, finding an effective pharmaceutical alternative for cornea transplantation is critical for the treatment of corneal endothelial dysfunction. In this study, we established a rabbit scratch model to investigate the effect of fibroblast growth factor 10 (FGF10) on corneal endothelial wound healing. Results showed that FGF10 injection accelerated the recovery of corneal transparency and increased the protein expression levels of ZO1, Na/K-ATPase and AQP-1. Moreover, FGF10 significantly inhibited the expression levels of endothelial-to-mesenchymal transition proteins and reduced the expression levels of the proinflammatory factors IL-1β and TNF-α in the anterior chamber aqueous humour. FGF10 also enhanced the Na/K-ATPase activity by enhancing mitochondrial function as a result of its direct interaction with its conjugate receptor. Thus, FGF10 could be a new pharmaceutical preparation as treatment for corneal endothelial dysfunction.
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http://dx.doi.org/10.1016/j.exer.2021.108517DOI Listing
February 2021

Clinical Characteristics and Outcomes of Short-term Repeat Corneal Transplantation.

Ocul Immunol Inflamm 2021 Feb 8:1-9. Epub 2021 Feb 8.

Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences , Qingdao, China.

Purpose: To explore the primary indications, direct causes and graft survival of short-term repeat keratoplasty.

Methods: An analytical cross-sectional study.

Results: The primary indications for keratoplasty were infectious keratitis (67.9%) and immune keratopathy (12.8%). The direct causes of graft failure were infectious ulcer (43.6%) and recurrence (30.8%). Pre-operative hypopyon (P = 0.017, HR = 1.757) and use of decellularized porcine corneas as grafts (P = 0.005, HR = 2.690) were independent risk factors for graft failure. The Kaplan-Meier analysis revealed that the 1-year survival rate of 88 repeat grafts was 85.2%, and the 5-year survival rate was 79.2%. The survival rates of the repeat grafts were significantly higher than those of the first grafts (P < 0.001).

Conclusion: Infectious keratitis is the most common indication of short-term graft failure and is often failed by infectious ulcer and keratitis recurrence. While the decellularized porcine cornea is a promising application in developing countries, certain difficulties, such as graft melting, remain unresolved.
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http://dx.doi.org/10.1080/09273948.2020.1838557DOI Listing
February 2021

c-Rel Is a Myeloid Checkpoint for Cancer Immunotherapy.

Nat Cancer 2020 May 18;1(5):507-517. Epub 2020 May 18.

Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.

Immunotherapy that targets lymphoid cell checkpoints holds great promise for curing cancer. However, a majority of cancer patients do not respond to this form of therapy. In addition to lymphoid cells, myeloid cells play essential roles in controlling immunity to cancer. Whether myeloid checkpoints exist that can be targeted to treat cancer is not well established. Here we show that c-Rel, a member of the nuclear factor (NF)-B family, specified the generation of myeloid-derived suppressor cells (MDSCs) by selectively turning on pro-tumoral genes while switching off anti-tumoral genes through a c-Rel enhanceosome. c-Rel deficiency in myeloid cells markedly inhibited cancer growth in mice, and pharmaceutical inhibition of c-Rel had the same effect. Combination therapy that blocked both c-Rel and the lymphoid checkpoint protein PD1 was more effective in treating cancer than blocking either alone. Thus, c-Rel is a myeloid checkpoint that can be targeted for treating cancer.
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http://dx.doi.org/10.1038/s43018-020-0061-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7808269PMC
May 2020

Individualized Corneal Patching for Treatment of Corneal Trauma Combined with Tissue Defects.

J Ophthalmol 2020 23;2020:8437479. Epub 2020 Nov 23.

Eye Hospital of Shandong First Medical University, Jinan, China.

Aim: To evaluate the efficacy of individualized corneal patching using a minimal graft for corneal trauma combined with tissue defects.

Methods: Fifteen eyes (15 patients) were enrolled in this study, including 8 eyes with corneal perforation induced by removal of metal foreign bodies, 5 eyes with corneal laceration resulting from metal trauma, and 2 eyes with pencil injuries to the cornea. The size, shape, and depth of the tissue defects were assessed. For corneal perforation or irregular tissue defects, if the diameter or length was ≥3.0 mm, traditional penetrating keratoplasty (PK) or lamellar keratoplasty (LK) was adopted; if the diameter or length was <3.0 mm, a conical or irregular patch consistent with the defects was used. The visual acuity, corneal status, and postoperative complications were observed during the follow-up.

Results: The diameter of corneal perforations was 1.0 mm in 2 eyes, 1.5 mm in 1 eye, 2.0 mm in 4 eyes, and 3.5 mm in 1 eye. During their PK procedures, a conical corneal graft was used in 7 eyes, while a traditional cylindrical graft was used in 1 eye. The other 7 eyes had corneal trauma combined with irregular tissue defects, which were full-thickness corneal defects in 5 eyes and lamellar defects in 2 eyes, all less than 3.0 mm in length. Thus, five eyes received PK, and 2 eyes received LK using an irregular wedge-shaped patch. The visual acuity increased greatly postoperatively, with mild corneal astigmatism. None of the patients developed immune rejection.

Conclusion: Individualized corneal patching with a minimal graft can save corneal materials, relieve corneal scars, gain a good visual prognosis, and avoid immune rejection in the treatment of corneal trauma combined with tissue defects.
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http://dx.doi.org/10.1155/2020/8437479DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7707941PMC
November 2020

Corneal calcification of acellular porcine corneal stroma following lamellar keratoplasty.

Acta Ophthalmol 2020 Nov 30. Epub 2020 Nov 30.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China.

Purpose: To describe the corneal calcification of acellular porcine corneal stroma (APCS) following lamellar keratoplasty (LKP) and identify risk factors.

Methods: Two cases of APCS calcification were evaluated by slit-lamp photography and anterior segment optical coherence tomography (AS-OCT). von Kossa staining and scanning electron microscope/energy-dispersive spectrometry (SEM/EDS) were performed on pathologic tissue. Associated graft and postoperative risk factors were analysed. Acellular porcine corneal stroma (APCS) cleanliness and element content after rinsing with sterilized water were observed by SEM/EDS and inductively coupled plasma mass spectrometry. Calcium metabolism-related proteins were analysed by protein mass spectrometry. Corneal epithelial defects and postoperative medications were reviewed.

Results: Two cases of APCS calcification occurred at 23 and 22 days postoperatively. Anterior segment optical coherence tomography (AS-OCT) and von Kossa staining demonstrated calcium deposition in the superficial stroma composed of calcium, phosphorus and oxygen conforming to the Ca/P ratio of hydroxyapatite. Phosphate crystals were present on the APCS surface and decreased with number of rinsing times. The phosphorus content of APCS was minimal after rinsing 10 times and avoiding excessive corneal swelling. Calcium metabolism-related proteins were downregulated in APCS. Patients with corneal calcification had 1-week postoperative corneal epithelial defects and were treated with three types of phosphorous eyedrops.

Conclusions: Acellular porcine corneal stroma (APCS) calcification occurs in the superficial corneal stroma about 1 month after LKP. The application of AS-OCT, von Kossa staining and SEM/EDS provides a basis for the clinical and pathological diagnosis of corneal calcification. The associated risk factors were mainly high phosphorus content and downregulated calcium metabolism-related proteins in APCS. Postoperative epithelial defects, inflammation and use of phosphorous eyedrops may promote corneal calcification.
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http://dx.doi.org/10.1111/aos.14665DOI Listing
November 2020

(DH) Exhibits Potent Anti-Proliferative Effects on Autoreactive CD4 T Cells and Ameliorates the Development of Experimental Autoimmune Uveitis.

Front Immunol 2020 8;11:575669. Epub 2020 Oct 8.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China.

Experimental autoimmune uveitis (EAU) is a CD4 T cell-mediated organ-specific autoimmune disease and has been considered as a model of human autoimmune uveitis. (DH) is a Chinese herbal medicine used in treating hepatitis. DH suppressed the production of inflammatory cytokines through the recruitment of myeloid-derived suppressor cells (MDSCs) to the liver. However, it remains elusive whether DH can directly regulate CD4 T cell biology and hence ameliorates the development of CD4 T cell-mediated autoimmune disease. In the current study, we found that DH extract significantly suppressed the production of pro-inflammatory cytokines by CD4 T cells. Further study showed that DH didn't affect the activation, differentiation, and apoptosis of CD4 T cells. Instead, it significantly suppressed the proliferation of conventional CD4 T cells both and . Mechanistic study showed that DH-treated CD4 T cells were partially arrested at the G2/M phase of the cell cycle because of the enhanced inhibitory phosphorylation of Cdc2 (Tyr15). In addition, we demonstrated that treatment with DH significantly ameliorated EAU in mice through suppressing the proliferation of autoreactive antigen specific CD4 T cells. Taken together, the current study indicates that DH-mediated suppression of CD4 T cell proliferation may provide a promising therapeutic strategy for treating CD4 T cell-mediated diseases.
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http://dx.doi.org/10.3389/fimmu.2020.575669DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7578250PMC
October 2020

Survey report on keratoplasty in China: A 5-year review from 2014 to 2018.

PLoS One 2020 15;15(10):e0239939. Epub 2020 Oct 15.

Department of Ophthalmology, Eye Hospital of Shandong First Medical University, State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Jinan, China.

To provide the general information on corneal transplantation (CT) in China, China Cornea Society designed a questionnaire on CT from 2014 to 2018 and entrusted it to 31 committee members for implementation of the survey nationwide. This article presents the results of the survey and compares the indicators used in the survey and those in the annual statistical report released by the Eye Bank Association of America (EBAA). The number of corneal transplantations completed by the 64 hospitals from 2014 to 2018 was respectively 5377, 6394, 7595, 8270 and 8980, totally 36,616 (22,959 male and 13,657 female). The five largest hospitals by the number of corneal transplantations completed 15,994 surgeries in total, accounting for 43.68% of all the surgeries performed in the 64 hospitals. The most common indication for corneal transplantations was corneal leukoma (7683, 20.98%), followed by bacterial keratitis (4209, 11.49%), corneal dystrophies (4189, 11.44%), keratoconus (3578, 9.77%) and corneal perforation (2839, 7.75%). The main surgical techniques were penetrating keratoplasty (PK) (19,896, 54.34%), anterior lamellar keratoplasty (ALK) (13,869, 37.88%). The proportion of PK decreased from 57.97% in 2014 to 52.88% in 2018 while the proportion of ALK increased from 36.04% in 2014 to 37.92% in 2018. The geographical distribution of keratoplasties performed in China is unbalanced. PK and ALK were the main techniques of CT and corneal leukoma, bacterial keratitis and corneal dystrophies were the main indications for CT in China.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0239939PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7561196PMC
November 2020

Hyperglycemia-reduced NAD biosynthesis impairs corneal epithelial wound healing in diabetic mice.

Metabolism 2021 01 11;114:154402. Epub 2020 Oct 11.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China. Electronic address:

Objective: Nicotinamide adenine dinucleotide (NAD) is an essential molecule participating in multiple physiological and pathophysiological processes. In diabetic cornea, the NAD-consuming enzyme SIRT1 was down-regulated and contributed to the delayed wound healing. However, the impact of hyperglycemia on corneal NAD biosynthesis remained elusive. This study was to investigate the relationship of NAD biosynthesis and the delayed corneal wound healing in diabetic mice.

Methods: Type 1 diabetes mellitus (DM) mice were induced by streptozotocin and corneal epithelial wound healing models were constructed by epithelial scraping. The NAD contents of corneal epithelium were measured using the NAD/NADH quantification kit. Expression of key enzymes involved in the NAD biosynthesis in type 1 DM mice and type 2 DM patients were analyzed. The nicotinamide phosphoribosyltransferase (NAMPT)-specific siRNA and the selective inhibitor FK866 were used to achieve the blockade of NAMPT, whereas exogenous NAD and its precursors were replenished to the corneal epithelial cells and DM mice.

Results: Hyperglycemia attenuated NAD content and NAMPT expression in the corneal epithelium of both type 1 DM mice and type 2 DM patients. Local knockdown of NAMPT by siRNA or FK866 consistently recapitulated the delayed corneal epithelial wound healing in normal mice. Moreover, NAD replenishment recovered the impaired proliferation and migration capacity by either FK866 or high glucose treatment in cultured corneal epithelial cells. Furthermore, in DM mice, NAD and its precursors nicotinamide mononucleotide and nicotinamide riboside also facilitated corneal epithelial and nerve regeneration, accompanied with the recovered expression of SIRT1 and phosphorylated EGFR, AKT, and ERK1/2 in epithelium and corneal sensitivity.

Conclusion: Hyperglycemia-reduced NAD biosynthesis and contributed to the impaired epithelial wound healing in DM mice. The replenishment of NAD and its precursors facilitated diabetic corneal wound healing and nerve regeneration, which may provide a novel therapeutic strategy for the treatment of diabetic corneal complications.
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http://dx.doi.org/10.1016/j.metabol.2020.154402DOI Listing
January 2021

Decellularized porcine cornea-derived hydrogels for the regeneration of epithelium and stroma in focal corneal defects.

Ocul Surf 2020 10 22;18(4):748-760. Epub 2020 Aug 22.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, China; Qingdao Eye Hospital of Shandong First Medical University, China. Electronic address:

Purpose: Hydrogels derived from decellularized tissues provide superior biocompatibility, tenability and tissue-specific extracellular matrix (ECM) components. Based on the preparation of decellularized porcine cornea (DPC), here we developed an injectable and transparent hydrogel for the regeneration of epithelium and stroma in focal corneal defects.

Methods: The DPC-derived hydrogel was prepared with N-cyclohexyl-N'-(2-morpholinethyl) carbodiimide metho-p-toluenesulfonate/N-hydroxysuccinimide (CMC/NHS) as cross-linkers. The characteristics of the hydrogel were analyzed and its cytocompatibility was assessed by Live/Dead and Cell Counting Kit (CCK)-8 assays. Immunofluorescence staining, quantitative PCR and Western blot analyses were performed to assess the relative protein and gene expression in corneal fibroblasts on hydrogel. The safety and efficiency of the hydrogel for repairing focal corneal defects in rabbit were measured by slit-lamp, anterior segment optical coherence tomography (AS-OCT), confocal microscopy and histological analyses.

Results: The DPC-derived hydrogel cross-linked with CMC/NHS assumed favorable transparency, exhibited distinct mechanical properties and preserved the ECM components of native porcine cornea (NPC). In vitro experiments showed that the hydrogel maintained the phenotype, supported the proliferation and promoted the ECM synthesis of corneal fibroblasts. When injected onto rabbit corneas, the hydrogel rapidly covered, solidified and formed a smooth surface on the focal defect. Corneal epithelium was fully regenerated within 3 days. The thickness of the corneal epithelium and stroma was restored at 12 weeks after surgery without significant inflammation or scar formation. Notably, the hydrogel showed no harmful effects on the resident stroma and endothelium.

Conclusions: The DPC-derived hydrogel may represent a promising biomaterial for corneal epithelial and stromal regeneration.
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http://dx.doi.org/10.1016/j.jtos.2020.07.020DOI Listing
October 2020

Real-time segmentation and tracking of excised corneal contour by deep neural networks for DALK surgical navigation.

Comput Methods Programs Biomed 2020 Dec 8;197:105679. Epub 2020 Aug 8.

Department of Computer Science, Stony Brook University, New York, US. Electronic address:

Objective: Corneal disease is one of the main causes of blindness for humans globally nowadays, and deep anterior lamellar keratoplasty (DALK) is a widely applied technique for corneal transplantation. However, the position of stitch points highly influences the success rate of such surgery, which would require accurate control and manipulation of surgical instruments.

Methods: In this paper, we present a deep learning framework for augmented reality (AR) based surgery navigation to guide the suturing in DALK. It can robustly track the excised corneal contour by semantic segmentation and the reconstruction of occlusion. We propose a novel optical flow inpainting network to recover the missing motion caused by occlusion. The occluded regions are detected by weakly supervised segmentation of surgical instruments and reconstructed by key frame warping along the completed optical flow. Then we introduce two types of loss function to adapt the inpainting network in the optical flow space.

Results: Our techniques are tested and evaluated by a number of real surgery videos from Shandong Eye Hospital in China. We compare our approaches with other typical methods in the corneal contour segmentation, optical flow inpainting and occlusion regions reconstruction. The tracking accuracy reachs 99.2% in average and PSNR reaches 25.52 for the reconstruction of the occluded frames.

Conclusion: From the experimental evaluations and user study, both the qualitative and quantitative results indicate that our techniques can achieve accurate detection and tracking of corneal contour under complex disturbance in real-time surgical scenes. Our prototype AR navigation system would be highly useful in clinical practice.
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http://dx.doi.org/10.1016/j.cmpb.2020.105679DOI Listing
December 2020

Clinical outcomes of double continuous suture in femtosecond laser-assisted lamellar keratoplasty for keratoconus.

Lasers Med Sci 2020 Aug 3. Epub 2020 Aug 3.

Shandong Eye Hospital, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, 372 Jingsi Road, Jinan, 250021, China.

To assess the surgical outcomes in patients who underwent femtosecond laser-assisted lamellar keratoplasty with double continuous suture for keratoconus, 100 patients (102 eyes) with keratoconus in advanced stages undergoing femtosecond laser-assisted lamellar keratoplasty in Shandong Eye Hospital were studied. In the management of keratoconus, 50 patients (52 eyes) received double continuous suture, and 50 patients (50 eyes) underwent interrupted suture. The follow-up duration was 1 year. Best-corrected visual acuity (BCVA), corneal astigmatism, cosmetic outcomes, and surgical complications were measured as outcome indicators. The epithelium healed at 3 ± 2 days and 4 ± 2 days in the double continuous suture group and the interrupted suture groups, respectively (P > 0.05). At 6 months after surgery, the average visual acuity was 20/125 and 20/100 (P > 0.05), and the average BCVA was 20/32 and 20/40 (P > 0.05), respectively. At 1 year after surgery, the average visual acuity was 20/63 and 20/80 (P > 0.05), and the average BCVA was 20/32 and 20/25 (P > 0.05), respectively; the mean curvature was 43.24 ± 5.15 D and 43.31 ± 5.58 D (P > 0.05), the mean astigmatism was 3.21 ± 1.74 D and 5.35 ± 1.37 D (P < 0.05).The looseness of sutures were found in 2 patients and 15 patients in both groups, respectively (P < 0.05). No postoperative infection or immune rejection occurred in either group during the follow-up. Comparing with the interrupted suture, using the continuous suture in femtosecond laser-assisted lamellar keratoplasty for keratoconus markedly limited the looseness of sutures with lesser corneal astigmatism and better visual quality postoperative.
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http://dx.doi.org/10.1007/s10103-020-03114-yDOI Listing
August 2020

The NLRP3 inflammasome regulates corneal allograft rejection through enhanced phosphorylation of STAT3.

Am J Transplant 2020 12 24;20(12):3354-3366. Epub 2020 Jun 24.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China.

The success of corneal transplantation is limited by allograft rejection, but the pathogenic mechanisms of this disease remain poorly defined. In this study, we showed that the NOD, LRR-and pyrin domain-containing protein3 (NLRP3) inflammasome-mediated interleukin-1β (IL-1β) production exacerbated corneal allograft rejection. Extracellular ATP contributed to the NLRP3 inflammasome-mediated IL-1β release, which in turn was preferentially skewed toward Th17 differentiation via enhanced phosphorylation of STAT3. Pharmacological inhibition of IL-1β/IL-6-STAT3 signaling significantly delayed corneal allograft rejection. Thus, the identification of NLRP3 inflammasome's key role sheds new light on the pathogenesis of corneal allograft rejection and opens a potential new avenue for treating or preventing corneal allograft rejection.
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http://dx.doi.org/10.1111/ajt.16071DOI Listing
December 2020

Laminin 511 Precoating Promotes the Functional Recovery of Transplanted Corneal Endothelial Cells.

Tissue Eng Part A 2020 11 28;26(21-22):1158-1168. Epub 2020 Jul 28.

Qingdao Eye Hospital of Shandong First Medical University, Qingdao, China.

Corneal endothelial dysfunction is a major cause of corneal blindness and is mainly treated by corneal transplantation. However, the global shortage of donor cornea hampers its application. Intracameral injection of cultured primary corneal endothelial cells (CECs) was recently confirmed in clinical trials. However, abnormal adhesion of the grafted CECs affects the application of this strategy. In this study, we explored if laminin 511 (LN511) improves the therapeutic function of the intracameral CEC injection for corneal endothelial dysfunction. To mimic the late stage of corneal endothelial diseases, intense scraping was developed to remove CECs and extracellular matrix of the posterior Descemet's membrane (DM) without DM removal in rabbits. Then, Dulbecco's phosphate-buffered saline (DPBS) and LN511 were intracamerally injected as the control and intervention groups, respectively. We found that the injected LN511 could settle and form a coating on the posterior surface of DM. After CEC transplantation, corneal clarity of rabbits in the LN511 group was rapidly recovered within 7 days, whereas the corneal recovery took 14 days in the DPBS group. Corneal thickness of LN511 group decreased to 413.3 ± 20.8 μm 7 days after operation, which was significantly lower than 1086.3 ± 78.6 μm of DPBS group ( < 0.01). Moreover, for the grafted CECs, LN511 promoted the rapid adhesion, tight junction formation, and expression of Na/K-ATPase and ZO-1. analysis revealed that the functions of LN511 on the cultured human CECs mechanistically depended on the cell density and the nuclear-cytoplasmic translocation of the Yes-associated protein. Our study demonstrated that LN511 precoating promoted the adhesion of the transplanted CECs and enhanced the functional regeneration of the corneal endothelium. Thus, our data suggested that the strategy of LN511 precoating and CECs' intracameral injection could be a potential method for the therapy of corneal endothelial dysfunction. Impact statement Intracameral injection of cultured corneal endothelial cells (CECs) is a potential alternative therapy for corneal endothelial dysfunction and has been proven to be effective in clinical trials. However, abnormal adhesion of the grafted CECs affects its application. In this study, intense scraping was developed to remove CECs and extracellular matrix of the posterior Descemet's membrane (DM) without DM removal for the therapy of late stage of corneal endothelial diseases. Laminin 511 was intracamerally injected to form a coating, improve the posterior DM, enhance the adhesion of the grafted CECs, and promote the functional regeneration of CEC transplantation through Yes-associated protein signaling.
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http://dx.doi.org/10.1089/ten.TEA.2020.0047DOI Listing
November 2020

Clinical Characteristics and Visual Outcomes of Patients Hospitalized for Ocular Trauma in Shandong Province, China.

J Ophthalmol 2020 15;2020:5826263. Epub 2020 Apr 15.

Shandong Eye Hospital, Shandong Eye Institute, Shandong First Medical University, Shandong Academy of Medical Sciences, Jinan 250021, China.

Purpose: To analyze the clinical characteristics and visual prognoses of patients with ocular trauma treated in Shandong Eye Hospital.

Methods: The inpatient data of patients with eye injuries hospitalized in our institution from January 2014 to December 2018 were retrospectively reviewed, including demographic information, types of trauma, causes of injury, treatment, and initial and final visual acuities.

Results: A total of 1,425 patients (1,622 eyes), aged 39.5 ± 18.5 years, were included. The ratio of male to female was 5.3 : 1. Of the mechanical eye injuries, there were 490 (34.4%) open-globe injuries and 454 (31.9%) closed-globe injuries. Nonmechanical eye injuries had 426 patients (29.9%), while 55 patients (3.9%) had adnexal injuries. Over a half of the traumas were work-related (51.1%, 728 patients). Most patients were treated with surgical intervention (1,404 eyes, 87.9%). There were significant differences in the final visual acuities between open-globe injuries and closed-globe injuries ( < 0.001), as well as between mechanical injuries and nonmechanical injuries ( < 0.001). The final visual acuity was closely correlated with the initial visual acuity (Spearman's correlation coefficient = 0.618, < 0.001) and the OTS score (Spearman's correlation coefficient = 0.691, < 0.001).

Conclusion: Ocular trauma usually occurs in young and middle-aged men and in the workplace in Shandong Province. The proportion of nonmechanical injuries is high, and the prognosis is poor. A comprehensive understanding of the characteristics of ocular trauma is useful for blindness prevention and treatment.
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http://dx.doi.org/10.1155/2020/5826263DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7180499PMC
April 2020

NAD precursors protect corneal endothelial cells from UVB-induced apoptosis.

Am J Physiol Cell Physiol 2020 04 12;318(4):C796-C805. Epub 2020 Feb 12.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University and Shandong Academy of Medical Sciences, Qingdao, China.

Excessive exposure of the eye to ultraviolet B light (UVB) leads to corneal edema and opacification because of the apoptosis of the corneal endothelium. Our previous study found that nicotinamide (NIC), the precursor of nicotinamide adenine dinucleotide (NAD), could inhibit the endothelial-mesenchymal transition and accelerate healing the wound to the corneal endothelium in the rabbit. Here we hypothesize that NIC may possess the capacity to protect the cornea from UVB-induced endothelial apoptosis. Therefore, a mouse model and a cultured cell model were used to examine the effect of NAD precursors, including NIC, nicotinamide mononucleotide (NMN), and NAD, on the UVB-induced apoptosis of corneal endothelial cells (CECs). The results showed that UVB irradiation caused apparent corneal edema and cell apoptosis in mice, accompanied by reduced levels of NAD and its key biosynthesis enzyme, nicotinamide phosphoribosyltransferase (NAMPT), in the corneal endothelium. However, the subconjunctival injection of NIC, NMN, or NAD effectively prevented UVB-induced tissue damage and endothelial cell apoptosis in the mouse cornea. Moreover, pretreatment using NIC, NMN, and NAD increased the survival rate and inhibited the apoptosis of cultured human CECs irradiated by UVB. Mechanistically, pretreatment using nicotinamide (NIC) recovered the AKT activation level and decreased the BAX/BCL-2 ratio. In addition, the capacity of NIC to protect CECs was fully reversed in the presence of the AKT inhibitor LY294002. Therefore, we conclude that NAD precursors can effectively prevent the apoptosis of the corneal endothelium through reactivating AKT signaling; this represents a potential therapeutic approach for preventing UVB-induced corneal damage.
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http://dx.doi.org/10.1152/ajpcell.00445.2019DOI Listing
April 2020

Ocular surface repair using decellularized porcine conjunctiva.

Acta Biomater 2020 01 6;101:344-356. Epub 2019 Nov 6.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao 266000, China; Shandong Eye Hospital, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Jinan 250000, China. Electronic address:

The primary functions of the conjunctiva embody ocular surface protection and the maintenance of the tear film equilibrium. Severe conjunctival defects such as symblepharon may impair the integrity of ocular surface and cause loss of visual functions. Here we report the use of a decellularized porcine conjunctiva (DPC) for conjunctival reconstruction in rabbit models and in clinic. Our results show that the major xenoantigens are efficiently removed, while abundant matrix components and integrated microstructures are well preserved in the DPC. These characteristics provide mechanical support and favorable histocompatibility for repairing damaged conjunctiva. The DPC application has demonstrated enhanced transplant stability and improved epithelial regeneration in severe ocular surface damage comparing to those of amniotic membrane (AM), the most frequently applied matrix for ocular surface reconstruction nowadays. In order to test the DPC performance in clinic, three patients with pterygium and one patient with symblepharon underwent transplant with DPC. The grafts in all cases were completely re-epithelized and no graft melt or fibroplasia were observed. These results suggest that the strategy we developed is feasible and effective for conjunctival reconstruction and ocular surface repair. STATEMENT OF SIGNIFICANCE: In this study, we adopted an innovative approach to prepare decellularized porcine conjunctiva (DPC). The intricate conjunctiva-specific structures and abundant matrix components were preserved in DPC, which offers favorable mechanical properties for graft. DPC has shown positive effects in ocular surface repair, which has been proven particularly in a rabbit model with severe symblepharon. Reconstructed conjunctiva by DPC exhibited epithelial heterogeneity, extremely resembling that of native conjunctiva. In addition, results from clinical studies were encouraging for pterygium and symblepharon and clinical application of DPC is promising.
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http://dx.doi.org/10.1016/j.actbio.2019.11.006DOI Listing
January 2020

Rapid porcine corneal decellularization through the use of sodium N-lauroyl glutamate and supernuclease.

J Tissue Eng 2019 Jan-Dec;10:2041731419875876. Epub 2019 Sep 11.

Shandong Eye Hospital, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Jinan, China.

Corneal decellularization represents a promising alternative source of human donor with global shortage. Multiple methods have been developed for the preparation of decellularized porcine corneal stroma. However, most strategies relied on long-time treatment to facilitate the entry of detergents or nucleases, which may cause irreversible ultrastructural damage. Here, we developed a rapid decellularization method for porcine corneal stroma through the combined mild detergent sodium N-lauroyl glutamate (SLG) and supernuclease. Compared with traditional methods, the novel decellularization method allowed the efficient removal of xenoantigen DNA within 3 h, while retaining the ultrastructure, transparency, and mechanical properties of porcine corneas. When transplanted in rabbit model for 1 month, the decellularized porcine corneal grafts presented favorable transparency and biocompatibility without immune rejection. Therefore, the combined use of detergent SLG and supernuclease may serve as a promising method for the clinical use of decellularized porcine cornea.
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http://dx.doi.org/10.1177/2041731419875876DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6740050PMC
September 2019

Corneal Denervation Causes Epithelial Apoptosis Through Inhibiting NAD+ Biosynthesis.

Invest Ophthalmol Vis Sci 2019 08;60(10):3538-3546

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China.

Purpose: To determine if trigeminal innervations of the corneal epithelium maintains its integrity and homeostasis through controlling the nicotinamide adenine dinucleotide (NAD) content of this tissue.

Methods: Corneal denervation of C57BL/6 mice was induced by squeezing the nerve bundles that derive from the trigeminal ganglion and was confirmed by whole-mount corneal nerve staining and the sensation test. The apoptosis of the corneal epithelium was examined by TUNEL assay and annexin V/propidium iodide staining. NAD biosynthesis-related enzymes were analyzed by quantitative PCR, immunofluorescence staining, and Western blotting. FK866, an inhibitor of nicotinamide phosphoribosyltransferase (NAMPT), exogenous nicotinamide mononucleotide (NMN), and NAD+ were used to evaluate the effect of NAD+ on the apoptosis of cultured corneal epithelial cells and epithelial detachment in denervated mice. Protein expression that related to apoptosis and phosphorylation were analyzed by Western blotting.

Results: The denervated mice showed spontaneous corneal epithelial detachment and cell apoptosis accompanied with impaired epithelial NAD+ contents due to low levels of NAMPT. Similarly, inhibition of NAMPT recapitulated epithelial detachment as in denervated mice and induced apoptosis in cultured corneal epithelial cells. The replenishment of NMN or NAD+ partially slowed down corneal nerve fiber degeneration, reduced the epithelial defect in denervated mice, and improved apoptosis induction in FK866-treated cells by restoring the activation levels of SIRT1, AKT, and CREB.

Conclusions: Corneal denervation lowered epithelial NAD+ contents through reducing the expression of NAMPT and caused cell apoptosis and epithelial defects, suggesting that corneal innervations contribute to epithelial homeostasis by regulating NAD+ biosynthesis.
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http://dx.doi.org/10.1167/iovs.19-26909DOI Listing
August 2019

Clinical features and serial changes of Acanthamoeba keratitis: an in vivo confocal microscopy study.

Eye (Lond) 2020 02 10;34(2):327-334. Epub 2019 Jul 10.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China.

Purpose: To observe the clinical features and serial changes of Acanthamoeba keratitis (AK) during medical treatment by using confocal microscopy.

Methods: Thirty-seven patients (37 eyes) diagnosed with AK were included in this study. Confocal microscopy was used to observe the morphology, distribution, and density of Acanthamoeba cysts before and after medication. The differences between cysts and inflammatory cells were identified.

Results: Acanthamoeba cysts were detected at a rate of 94.6% (35/37) by repeated confocal microscopic examinations. The cysts consisting of a lowly light-reflective wall and a high-refractive nucleus, showed cluster or chain distribution in the corneal stroma, which was different from inflammatory cells. After medical therapy, the nucleus of cysts or peripheral corneal tissue gradually dissolved to a hollow configuration. Some of the hollow cysts existed for up to 6 months. The quantity of cysts increased after 1-2 weeks of medication in 23 patients (62.1%), and then began to decrease in 13 patients (35.1 %) who were responsive to anti-amoebic treatment.

Conclusion: Acanthamoeba cysts have many typical clinical features that can be identified by confocal microscopy, which may serve as a valuable tool to guide clinical evaluation and treatment of AK.
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http://dx.doi.org/10.1038/s41433-019-0482-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7002551PMC
February 2020

Factors Affecting Myopic Regression after Laser in Situ Keratomileusis and Laser-Assisted Subepithelial Keratectomy for High Myopia.

Semin Ophthalmol 2019 1;34(5):359-364. Epub 2019 Jul 1.

a Refractive Surgery Department, Shandong Eye Hospital, Shandong Eye Institute, Shandong Academy of Medical Sciences , Jinan , China.

To evaluate the factors affecting the amount of myopic regression after laser refractive surgery for high myopia, we retrospectively reviewed medical records of 1591 eyes with high myopia treated by laser in situ keratomileusis or laser-assisted subepithelial keratectomy at Shandong Eye Hospital between January 2008 and December 2012. Thirty-five eyes suffering a postoperative myopic shift of manifest spherical equivalent (MSE) of 0.25 diopter (D) or greater were included in this study. The mean refractive error was -9.34 ± 1.89 D before surgery, and the mean regression was -1.22 ± 0.70 D. Correlation analysis and multiple regression analysis were performed to assess the factors that were associated with the refractive regression. The age displayed a negative correlation with the diopter of regression ( = -0.404, .016), while the optical zone diameter had a positive correlation with it ( = 0.406, .016). Explanatory variables relevant to the diopter of regression were age (partial regression coefficient B = -0.045, = .016) and optical zone diameter (partial regression coefficient B = 0.979, = .014). Advanced-age patients with small optical zones were more predisposed to a larger amplitude of regression after laser refractive surgery for high myopia.
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http://dx.doi.org/10.1080/08820538.2019.1632353DOI Listing
August 2019

Bladder drug mirabegron exacerbates atherosclerosis through activation of brown fat-mediated lipolysis.

Proc Natl Acad Sci U S A 2019 05 13;116(22):10937-10942. Epub 2019 May 13.

Department of Microbiology, Tumor and Cell Biology, Karolinska Institute, 171 77 Stockholm, Sweden;

Mirabegron (Myrbetriq) is a β3-adrenoreceptor agonist approved for treating overactive bladder syndrome in human patients. This drug can activate brown adipose tissue (BAT) in adult humans and rodents through the β3-adrenoreceptor-mediated sympathetic activation. However, the effect of the mirabegron, approved by the US Food and Drug Administration, on atherosclerosis-related cardiovascular disease is unknown. Here, we show that the clinical dose of mirabegron-induced BAT activation and browning of white adipose tissue (WAT) exacerbate atherosclerotic plaque development. In apolipoprotein E () and low-density lipoprotein (LDL) receptor () mice, oral administration of clinically relevant doses of mirabegron markedly accelerates atherosclerotic plaque growth and instability by a mechanism of increasing plasma levels of both LDL-cholesterol and very LDL-cholesterol remnants. Stimulation of atherosclerotic plaque development by mirabegron is dependent on thermogenesis-triggered lipolysis. Genetic deletion of the critical thermogenesis-dependent protein, uncoupling protein 1, completely abrogates the mirabegron-induced atherosclerosis. Together, our findings suggest that mirabegron may trigger cardiovascular and cerebrovascular diseases in patients who suffer from atherosclerosis.
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http://dx.doi.org/10.1073/pnas.1901655116DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6561204PMC
May 2019

TIPE2 Suppresses Pseudomonas aeruginosa Keratitis by Inhibiting NF-κB Signaling and the Infiltration of Inflammatory Cells.

J Infect Dis 2019 08;220(6):1008-1018

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China.

Background: The role of tumor necrosis factor α (TNF-α) induced protein 8-like-2 (TIPE2) in Pseudomonas aeruginosa (PA) keratitis was explored.

Methods: Eight-week-old TIPE2 knockout (TIPE2-/-) C57BL/6 mice and their wild-type (WT) littermates were used. Corneal disease was graded at 1, 2, and 3 days postinfection, and slit lamp, clinical score, histopathology, and immunostaining were performed in the infected corneas. The corneas were harvested, and messenger ribonucleic acid (mRNA) levels of TNF-α, interleukin-1β (IL-1β), and interleukin-6 (IL-6) were tested. Enzyme-linked immunosorbent assay (ELISA) determined the protein levels, and nuclear factor κ-light-chain-enhancer of activated B cell (NF-κB) signaling molecules were tested by Western blot. In vitro human corneal epithelial cells (HCECs) were used to determine the relationship between TIPE2 and TAK1. The HCECs were treated with TIPE2 short hairpin ribonucleic acid (shRNA) and lipopolysaccharide (LPS) to test the NF-κB signaling molecules by Western blot.

Results: Pseudomonas aeruginosa infection induced a decreased expression of TIPE2 in mouse corneas 2 days postinfection. Compared with the control group, TIPE2-deficient mice were susceptible to infection with PA and showed increased corneal inflammation. Reduced NF-κB signaling and inflammatory cell infiltration were required in the TIPE2-mediated immune modulation.

Conclusions: TIPE2 promoted host resistance to PA infection by suppressing corneal inflammation via regulating TAK1 signaling negatively and inhibiting the infiltration of inflammatory cells.
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http://dx.doi.org/10.1093/infdis/jiz246DOI Listing
August 2019

Nicotinamide inhibits corneal endothelial mesenchymal transition and accelerates wound healing.

Exp Eye Res 2019 07 24;184:227-233. Epub 2019 Apr 24.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, China. Electronic address:

Corneal endothelial cells (CECs) maintain the clarity of the cornea through the barrier and pump function. Ex vivo culture or injury may cause corneal endothelial-mesenchymal transition (EnMT) and lead to loss of function. In this study, we explored the effects of nicotinamide (NIC) on the wound healing of rabbit corneal endothelium and the proliferation, migration, and EnMT of cultured human CEC lines. The animal results showed that corneal clarity was rapidly recovered within seven days through topical application of NIC in the rabbits with mechanical injury of the corneal endothelium, while the control corneas remained edematous and cloudy. Whole-mounted corneal staining found the expressions of Na/K-ATPase, aquaporin-1, and zonula occludens-1 were mainly localized to the boundaries of regenerated endothelium in NIC-treated eyes, in contrast to the scattered staining in vehicle-treated eyes. Interestingly, we found that NIC application inhibited the expression of typical EnMT marker alpha-smooth muscle actin, which appeared in the rabbit corneal endothelial wound healing. In vitro, NIC promoted the proliferation, but not the migration, of cultured human CECs. Moreover, NIC effectively inhibited transforming growth factor beta-1-induced corneal EnMT and decreased the levels of EnMT regulators snail and slug. Therefore, our study indicates that NIC enhances corneal endothelial wound healing through the promotion of proliferation and the inhibition of EnMT, which may provide a potential pharmaceutical agent for treating corneal endothelial dysfunction.
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http://dx.doi.org/10.1016/j.exer.2019.04.012DOI Listing
July 2019

Blood Vessels and Lymphatic Vessels in the Cornea and Iris After Penetrating Keratoplasty.

Cornea 2019 Jun;38(6):742-747

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of Medical Sciences, Qingdao, China.

Purpose: To detect early growth of blood and lymphatic vessels in the mouse cornea and iris after penetrating keratoplasty.

Methods: Penetrating keratoplasty was performed with C57BL/6 mice as donors and BALB/c mice as recipients. Graft transparency and neovascularization were examined by slit-lamp microscopy. Whole mounts of the cornea and iris were processed for detection of the outgrowth of blood and lymph vessels.

Results: On day 3 after surgery, all corneal grafts were slightly edematous, and blood vessels in the corneoscleral limbus dilated. LYVE-1 lymphatic vessels and CD31 blood vessels were distributed in the peripheral cornea. In the iris, the density of blood vessels increased, and LYVE-1 cells nearly vanished. On day 7, the grafts became opaque, and blood vessels grew into the recipient bed. A great quantity of lymph vessels invaded the cornea. LYVE-1 arborescent cells were found around the lymphatic vessels. In the iris, blood vessels became bulky and stiff, and arborescent LYVE-1 cells increased in number. On day 14, corneal neovascular regression and graft clarity were found. Lymphatic vessels regressed more slowly than blood vessels in the cornea. In the iris, blood vessels remained coarse. Increasing arborescent LYVE-1 cells were also noted in the ciliary body.

Conclusions: Our findings suggest that the iris-ciliary body could amplify immune signals and in part promote initiation of immune rejection after keratoplasty by providing a pathway for macrophages, which might participate in corneal lymphangiogenesis.
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http://dx.doi.org/10.1097/ICO.0000000000001922DOI Listing
June 2019

Rapid Differentiation of Multi-Zone Ocular Cells from Human Induced Pluripotent Stem Cells and Generation of Corneal Epithelial and Endothelial Cells.

Stem Cells Dev 2019 04 5;28(7):454-463. Epub 2019 Mar 5.

1 Medical College, Qingdao University, Qingdao, China.

Eye is a complex organ with a highly specialized tissue structure. The establishment of human pluripotent stem cells (hPSCs) has allowed the simulation of eye development in vitro. Most differentiation works of hPSC-derived ocular cells focus on a single, tissue-specific lineage, however, that faces difficulty in reflecting the complexity of eye development. Recently, the generation of a self-formed ectodermal autonomous multi-zone of ocular cells availably mimics the process of whole-eye development. In this study, we developed a rapid defined method to induce the differentiation of multi-zone ocular cells (MZOCs) from human induced pluripotent stem cells, which specifically experienced the key progenitor stages of anterior neuroectoderm and eye field stem cells by a 2.5-dimensional culture. These differentiated cell types spanned neural retina, retinal pigment epithelium, surface ectoderm, and neural crest and lens cells. In addition, the surface ectoderm zone of MZOCs could be mechanically isolated and induced into corneal epithelial cells, and the isolated neural crest zone could be directed into corneal endothelial cells. This in vitro differentiation process vividly mimics the development of vertebrate eye, and it provides a promising model for the study of ocular morphogenesis, as well as an ideal resource of seed cells for corneal regenerative medicine.
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http://dx.doi.org/10.1089/scd.2018.0176DOI Listing
April 2019

Characteristics of New Onset Herpes Simplex Keratitis after Keratoplasty.

J Ophthalmol 2018 22;2018:4351460. Epub 2018 Oct 22.

Shandong Eye Hospital, Shandong Eye Institute, Shandong Academy of Medical Sciences, Jinan, China.

Purpose: To observe clinical characteristics and treatment outcomes of new onset herpes simplex keratitis (HSK) after keratoplasty.

Methods: Among 1,443 patients (1,443 eyes) who underwent keratoplasty (excluding cases of primary HSK) in Shandong Eye Hospital, 17 patients suffered postoperative HSK. The clinical manifestations, treatment regimens, and prognoses of the patients were evaluated.

Results: The incidence of new onset HSK after keratoplasty was 1.18%. Epithelial HSK occurred in 10 eyes, with dendritic epithelial infiltration in 6 eyes and map-like epithelial defects in 4 eyes. Nine eyes had lesions at the junction of the graft and recipient. Stromal necrotic and endothelial HSK occurred in 7 eyes, presenting map-shaped ulcers in the entire corneal graft and recipient (two eyes) or at the graft-recipient junction (five eyes). Confocal microscopy revealed infiltration of a large number of dendritic cells at the junction of the lesion and transparent cornea. All 10 eyes with epithelial lesions and two eyes suffering stromal lesions of ≤1/3 corneal thickness healed after systematic and local antiviral treatment. Best-corrected visual acuity and corneal graft transparency were restored. For stromal HSK with an ulcer of >1/3 corneal thickness, amniotic membrane transplantation was performed, and visual acuity and graft transparency decreased significantly.

Conclusion: New onset HSK after keratoplasty primarily resulted in epithelial and stromal lesion, involving both the graft and recipient. Effective treatments included antiviral medications and amniotic membrane transplantation. Delayed treatment may lead to aggravated graft opacification.
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http://dx.doi.org/10.1155/2018/4351460DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6217905PMC
October 2018

Rapamycin Nano-Micelle Ophthalmic Solution Reduces Corneal Allograft Rejection by Potentiating Myeloid-Derived Suppressor Cells' Function.

Front Immunol 2018 8;9:2283. Epub 2018 Oct 8.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of Medical Sciences, Qingdao, China.

Allograft rejection is the major cause of corneal allograft failure. Rapamycin (RAPA) has been reported as an effective and novel immunosuppressive agent for patients undergoing corneal transplantation. However, its high water insolubility and low bioavailability have strongly constrained its clinical application. In this study, we successfully developed a RAPA nano-micelle ophthalmic solution and found that corneal allograft survival in recipients treated with RAPA nano-micelle ophthalmic solution was significantly prolonged for more than 2 months, with less inflammatory infiltration, decreased production of pro-inflammatory factors, and elevated recruitment of myeloid-derived suppressor cells (MDSCs). MDSCs from mice treated with RAPA nano-micelle ophthalmic solution could significantly inhibit the proliferation of CD4T cells through increased expressions of inducible nitric oxidase (iNOS) and arginase-1 (Arg-1). The activity blockade of Arg-1 and iNOS pharmacologically reversed their immunosuppressive ability. Moreover, the effects of RAPA were antagonized by the administration of anti-Gr-1 antibody or by inhibiting the activity of iNOS pharmacologically. In addition, RAPA nano-micelle also effectively alleviated allograft rejection in high-risk rabbit penetrating keratoplasty (PKP) models with corneal vascularization. Collectively, our results demonstrate that RAPA nano-micelle ophthalmic solution could improve the immunosuppressive activity of MDSCs through elevated expression of Arg-1 and iNOS, which highlights the possible therapeutic applications of RAPA against corneal allograft rejection.
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http://dx.doi.org/10.3389/fimmu.2018.02283DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6186809PMC
September 2019

Different Effects of Pro-Inflammatory Factors and Hyperosmotic Stress on Corneal Epithelial Stem/Progenitor Cells and Wound Healing in Mice.

Stem Cells Transl Med 2019 01 9;8(1):46-57. Epub 2018 Oct 9.

State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of Medical Sciences, Qingdao, People's Republic of China.

Chronic inflammation and severe dry eye are two important adverse factors for the successful transplant of cultured limbal stem cells. The aim of this study was to investigate the effects of inflammation and hyperosmotic stress (a key pathological factor in dry eye) on corneal epithelial stem cells (CESCs) and corneal epithelial wound healing. We observed that the CESCs exhibited significant morphological changes when treated with interleukin-1 beta (IL-1β), tumor necrosis factor alpha (TNF-α), or hyperosmotic stress. Colony-forming efficiency or colony-forming size was decreased with the increasing concentrations of IL-1β, TNF-α, or hyperosmotic stress, which was exacerbated when treated simultaneously with pro-inflammatory factors and hyperosmotic stress. However, the colony-forming capacity of CESCs recovered more easily from pro-inflammatory factor treatment than from hyperosmotic stress treatment. Moreover, when compared with pro-inflammatory factors treatment, hyperosmotic stress treatment caused a more significant increase of apoptotic and necrotic cell numbers and cell cycle arrest in the G2/M phase. Furthermore, the normal ability of corneal epithelial wound healing in the mice model was suppressed by both pro-inflammatory factors and hyperosmotic stress treatment, and especially severely by hyperosmotic stress treatment. In addition, inflammation combined with hyperosmotic stress treatment induced more serious epithelial repair delays and apoptosis in corneal epithelium. Elevated levels of inflammatory factors were found in hyperosmotic stress-treated cells and mice corneas, which persisted even during the recovery period. The results suggested that pro-inflammatory factors cause transient inhibition, while hyperosmotic stress causes severe apoptosis and necrosis, persistent cell cycle arrest of CESCs, and severe corneal wound healing delay. Stem Cells Translational Medicine 2019;8:46-57.
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http://dx.doi.org/10.1002/sctm.18-0005DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6312447PMC
January 2019

Autocrine IL-1β mediates the promotion of corneal neovascularization by senescent fibroblasts.

Am J Physiol Cell Physiol 2018 11 29;315(5):C734-C743. Epub 2018 Aug 29.

Shandong Provincial Key Lab of Ophthalmology, Shandong Eye Institute, Shandong Academy of Medical Sciences , Qingdao, Shandong , China.

Our previous study has confirmed that senescent fibroblasts promote corneal neovascularization (CNV) partially via the enhanced secretion of matrix metalloproteases (MMPs). However, the regulation of MMP expression in senescent fibroblasts remained unclear. In this study, we identified that the expression and secretion levels of interleukin-1β (IL-1β) were significantly upregulated in senescent human corneal fibroblasts than that in normal fibroblasts. Moreover, compared with vehicle-pretreated senescent fibroblasts, IL-1β pretreatment enhanced the expression of angiogenic factors but reduced the expression of angiostatic factors in senescent fibroblasts. When cocultured with human umbilical vein endothelial cells, IL-1β-pretreated senescent fibroblasts more strongly promoted their proliferation, migration, and tube-formation capacities than the vehicle-controlled senescent fibroblasts. In addition, either interleukin-1 receptor antagonist or anti-IL-1β neutralization completely inhibited the promotion of senescent fibroblasts in vascular tube formation in vitro and CNV in vivo. Therefore, we concluded that autocrine IL-1β mediated the promotion of senescent fibroblasts on corneal neovascularization.
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http://dx.doi.org/10.1152/ajpcell.00205.2017DOI Listing
November 2018