The role of fibronectin in corneal wound healing explored by a physician–scientist

For the past 30?years, I have worked as a physician-scientist in both the clinic and laboratory setting at a number of university medical schools. Encountering patients in the clinic for whom treatment was not available led me to the laboratory in an attempt to develop the appropriate treatment for future patients. The main focus of my translational research has been the role of fibronectin in corneal epithelial wound healing and the development of fibronectin eyedrops for the treatment of patients with persistent corneal epithelial defects. An extension of this research led to the development of eyedrops containing the synthetic peptide proline-histidine-serine-arginine-asparagine (PHSRN), which corresponds to the second cell-binding site of fibronectin. My clinical experience with fibronectin eyedrops also prompted me to examine the role of the sensory neurotransmitter substance P and insulin-like growth factor-1 (IGF-1) in corneal wound healing, leading to the development of eyedrops containing peptides derived from these agents (peptides FGLM-amide and SSSR, respectively). Although the path from the laboratory to the clinic in these instances has been relatively short, the time required to establish the newly identified treatment modalities in the wider community has been long. In this review, I relate the trajectory of my translational research career.     

The role of protein tyrosine phosphorylation in the cell–cell junctions and intercellular permeability of post-confluent bovine corneal epithelial cells

PurposeTo evaluate the role of protein tyrosine phosphatase (PTP) in controlling the integrity of cell–cell junction and intercellular permeability in postconfluent bovine corneal epithelial cells.MethodsConfluent cultures of bovine corneal epithelial cells were treated with different concentrations of general phosphate inhibitors andsodium orthovanadate for varying periods. An MTS assay was used to confirm no cellular death under the treatment profile. Immunocytochemical (ICC) analysis was performed to demonstrate protein tyrosine phosphorylation after treatment with sodium orthovanadate, and the effect of sodium orthovanadate on junctional proteins such as p120, α-catenin, occludin, ZO-1, and ZO-2. Western blot analysis was used to analyze the changes in p120, α-catenin, occludin, ZO-1, and ZO-2 after treatment. Paracellular permeability was evaluated by transepithelial electrical resistance (TER).ResultsDuring the whole course of treatment, no significant cellular death was noticed. Dose- and time-dependent effects of sodium orthovanadate on protein tyrosine phosphorylation were confirmed by ICC. ICC also demonstrated the dose- and time-dependent effect of sodium orthovanadate on the disruption of p120, α-catenin, occludin, ZO-1, and ZO-2. However, results of Western blot analysis showed no change in the expression levels of p120, α-catenin, occludin, ZO-1, and ZO-2. Dose- and time-dependent increase of paracellular permeability was evaluated by TER.ConclusionInhibition of protein tyrosine phosphatase activity can increase protein tyrosine phosphorylation. A dose- and time-dependent release of cell–cell contacts and increased transepithelial permeability were found in postconfluent culture of bovine corneal epithelial cells.     

Gene–gene interaction of CFH,ARMS2, and ARMS2/HTRA1 on the risk of neovascular age-related macular degeneration and polypoidal choroidal vasculopathy in Chinese population

Purpose

To evaluate the association and interaction of five single-nucleotide polymorphisms (SNPs) in three genes (CFH, ARMS2, and ARMS2/HTRA1) with neovascular age-related macular degeneration (nAMD) and polypoidal choroidal vasculopathy (PCV) in Chinese population.

Methods

A total of 300 nAMD and 300 PCV patients and 301 normal subjects participated in the present study. The allelic variants of rs800292, rs2274700, rs3750847, rs3793917, and rs1065489 were determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Gene–gene interactions were evaluated by the data mining approach multifactor-dimensionality reduction (MDR) method.

Results

The risk alleles of CFH rs800292, rs2274700, ARMS2 rs3057847, and ARMS2/HTRA1 rs3793917 showed significant difference between nAMD or PCV patients and controls (all P<0.01). The homozygosity of risk alleles for rs800292, rs2274700, rs3750847, and rs3793917 were significantly different between nAMD patients and controls (all P<0.01), and predisposed to PCV patients (all P<0.01). After cross-validation consistency (CVC) and permutation tests, the two-locus model rs2274700_rs3750847 has a balanced accuracy of 64.37% in predicting nAMD disease risk. The one-marker model, rs3750847, and two-locus model rs2274700_rs3750847 has a balanced accuracy of 66.07% and 65.89% in predicting PCV disease risk, respectively. Furthermore, CFH rs1065489 did not show significant association with nAMD (P>0.01), but was strongly associated with PCV in Chinese patients (P<0.001).

Conclusions

In this study, we found that the interaction of ARMS2 and ARMS2/HTRA1 is significantly associated with nAMD, and the interaction of CFH and ARMS2 is pronounced in PCV development in Chinese population.     

Progression and associated factors of lacquer cracks/patchy atrophies in high myopia: the Beijing Eye Study 2001–2011

Graefe's Archive for Clinical and Experimental Ophthalmology - To assess the development and progression of lacquer cracks/patchy atrophies (LCs/PAs) in high myopia. The case control study...     

Figure 1 CD40 , CD68 & CD40L expression in the epidermis and subcutaneous tissue of erythema nodosum (×200) A: CD40+ (red) (AEC) in epidermis；B: CD68＋ (black) (BCIP/NBT) in subcutaneous tissue；C: CD40L (red) (AEC) in subcutaneous tissue Bioinformatics analysis and construction of eukaryotic expression plasmid of Cx50 V64G mutation

AIM: To construct and analyze eukaryotic expression plasmid inserted by Cx50 with V64G mutation through bioinformatics software. METHODS: The full coding domain sequence of Cx50 with V64G mutation was acquired from the blood of patients with cataract and was cloned into pcDNA3.1/Amp (+).The constructed plasmid was identified with PCR , enzyme digestion and sequencing. The analysis of Cx50 with V64G mutation was performed with bioinformatics software. RESULTS: Cx50 with V64G mutation was successfully amplified and its eukaryotic expression plasmid was constructed. Valine-64 is well conserved in the first extracellular loop of connexin 50 in different species and also in different human α-type gap junctional proteins. CONCLUSION: The successive reconstruction and verification of eukaryotic expression plasmid containing Cx50 with V64G mutation established the foundation for further studying the mechanism of cataract.