PURPOSE. To evaluate spontaneous eye blink patterns and their correlations with clinical tests in dry eye disease (DED). METHODS. Twenty-five DED patients and 25 healthy subjects were included in this prospective case-controlled study. Clinical evaluations included the Ocular Surface Disease Index (OSDI), lipid layer thickness (LLT), spontaneous eye blink pattern analysis, fluorescein tear film breakup time (FBUT), and so on. Eye blinks were recorded for 20 seconds with a high-speed camera. Eye blink patterns were divided into the following five phases: the eyelid closing phase (ECP), eyelid closed phase (CDP), early opening phase (EOP), late opening phase (LOP), and interblink intervals (IBI). The correlations between blink parameters and clinical tests were analyzed. RESULTS. Compared with the control group, mean ECP, CDP, and EOP were significantly longer in DED patients (P < 0.001, P ¼ 0.029, and P < 0.001, respectively). DED patients also had significantly shorter LOP and blink intervals (both P < 0.001) and more partial blinks as compared with control subjects (P ¼ 0.001). FBUT was negatively correlated with ECP (r ¼ À0.618, P¼0.001) and the number of partial blinks (r ¼ À0.413, P ¼ 0.040). There was a positive correlation between OSDI and the number of partial blinks (r ¼ 0.446, P ¼ 0.026). The LLT coefficient of variation (LLT-CV) also showed a positive correlation with ECP, CDP, and LOP

Abstract. The aim of the present study was to investigate the associations between vasculogenic mimicry (VM) and zinc finger E-box binding homeobox 1 (ZEB1) in bladder cancer. VM structure and ZEB1 expression were analyzed by cluster of differentiation 34/periodic acid Schiff (PAS) double staining and immunohistochemical staining in 135 specimens from patients with bladder cancer, and a further 12 specimens from normal bladder tissues. Three-dimensional (3-D) culture was used to detect VM formation in the bladder transitional cancer cell lines UM-UC-3 and J82, and the immortalized human bladder epithelium cell line SV-HUC-1 in vitro. ZEB1 expression in these cell lines was compared by reverse transcription-quantitative polymerase chain reaction and western blot assays. In addition, small interfering RNA was used to inhibit ZEB1 in UM-UC-3 and J82 cells, followed by 3-D culturing of treated cell lines. As a result, VM was observed in 31.1% of specimens from bladder cancer tissues, and cases with high ZEB1 expression accounted for 60.0% of patients with bladder cancer. In addition, ZEB1 expression was closely associated with VM (r=0.189; P<0.05), and also increased as the grade and stage of the tumor developed. In an in vitro assay, UM-UC-3 and J82 cells exhibited VM formation, however, SV-HUC-1 did not. Furthermore, VM-forming cancer cell lines UM-UC-3 and J82 exhibited higher ZEB1 expression. Notably, VM formation was inhibited following knockdown of ZEB1. In conclusion, ZEB1 may be associated with VM in bladder cancer and serve an important role in the process of VM formation. However, its detailed mechanism requires further study.

Purpose We evaluate the antimicrobial effect of toluidine blue O (TBO)-mediated photodynamic antimicrobial chemotherapy (PACT) on Staphylococcus epidermidis and Staphylococcus aureus isolated from ocular surface infection. Methods We selected 24 strains of bacteria for this study. The antimicrobial effect of different TBO concentrations, light irradiation, and duration were evaluated. After determining the optimal PACT parameters, four experimental groups were included: Group 1, TBO alone (T+L−); Group 2, light-emitting diode (LED) irradiation alone (T−L+); Group 3, TBO–LED irradiation combination (T+L+); and Group 4, no treatment group (T−L−). The antibacterial effect of PACT was evaluated with the colony survival fraction (SF) methods. Results The antibacterial effect of PACT on S. epidermidis and S. aureus was dose-dependent to light irradiation and TBO concentration. The optimal PACT parameters were a TBO concentration of 60 μM, light irradiation of 5.27 mW/cm 2 , and an irradiation duration of 30 minutes. In group 1, 60 μM TBO without irradiation did not show any antibacterial effect on S. epidermidis (1.48E7 ± 1.5E6 colony-forming units [CFU]/mL) or S. aureus (1.45E7 ± 9E5 CFU/mL). In group 2, irradiation alone with 5.27 mW/cm 2 did not modify bacterial growth ( S. epidermidis , 1.49E7 ± 1.43E6; S. aureus, 1.5E7 ± 1.2E6). In group 3, after treatment, bacteria density dropped to 4000 ± 1000 and 3E5 ± 1E5 CFU/mL in S. epidermidis and S. aureus groups, respectively ( P < 0.001, P = 0.030). In group 4, there was uniform bacterial growth ( S. epidermidis , 1.51E7 ± 1.5E6; S. aureus, 1.48E7 ± 1.5E6) before and after treatment. Conclusions TBO-mediated PACT had an antibacterial efficacy in vitro on S. epidermidis and S. aureus isolated from ocular surface infection. Translational Relevance As TBO-mediated PACT has a strong antibacterial effect to S. epidermidis and S. aureus in vitro, this approach may assist in the treatment of ocular surface infectious diseases.

In order to study Staphylococcus epidermis and Staphylococcus aureus in vitro viability after the exposure to ultraviolet (UV) light and riboflavin, twelve strains of Staphylococcus epidermis and twelve strains of Staphylococcus aureus were isolated from patients with bacterial keratitis. The growth situation of Staphylococcus epidermidis and Staphylococcus aureus under different experimental conditions was qualitatively observed. The number of colonies surviving bacteria was counted under different UV light power and different exposure time. The experiment showed that there was no inhibition effect on the growth of bacteria using riboflavin alone. In UV alone group and UV-riboflavin group, inhibition effect on the bacteria growth was found. The UV-riboflavin combination had better inhibition effect on bacteria than UV irradiation alone. The amount of bacteria in the UV-riboflavin group was decreased by 99.1%~99.5% and 54.8%~64.6% in the UV alone group, when the UV light power was 10.052 mW/cm2 and the irradiation time was 30 min. Moreover, with the increase of the UV power or irradiation time, the survival rates of bacteria were rapidly reduced. Compared with Staphylococcus aureus, Staphylococcus epidermis was more easily to be killed under the action of UV light combined with riboflavin.

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Corneal regeneration has become a prominent study area in recent decades. Because the corneal stroma contributes about 90% of the corneal thickness in the corneal structure, corneal stromal regeneration is critical for the treatment of cornea disease. Numerous materials, including deacetylated chitosan, hydrophilic gel, collagen, gelatin methacrylate (GelMA), serine protein, glycerol sebacate, and decellularized extracellular matrix, have been explored for keratocytes regeneration. GelMA is one of the most prominent materials, which is becoming more and more popular because of its outstanding three-dimensional scaffold structure, strong mechanics, good optical transmittance, and biocompatibility. This review discussed recent research on corneal stroma regeneration materials and related GelMA.

The objective of this study was to evaluate the effect of toluidine bluemediated photodynamic therapy on experimental bacterial keratitis in rabbits. Methods: Bacterial keratitis was induced in rabbits by the injection of 200 μl Staphylococcus aureus (S. aureus) solution into the anterior stroma of the right cornea. Rabbits were randomly divided into four groups: toluidine blue O and red light (TBOR), levofloxacin eye drops (LEV), the combination of TBOR and LEV (TBOR + LEV), and a control group. Clinical manifestations, histopathology, and transmission electron microscopy were analyzed at various time points. Results: Conjunctival injection and surface area of the corneal ulcer in the TBOR group and the TBOR + LEV group showed significant improvement from baseline after 7 days of treatment. Compared to baseline, the depth of corneal infiltration was decreased at day 14 in the TBOR and TBOR + LEV groups. Microscopic analysis of the TBOR and TBOR + LEV groups showed that the structure of each layer was intact, and there were no inflammatory cells in the corneal stroma. Additionally, IL-1β and TNF-α were highly expressed in the LEV and control groups but were lower in the TBOR and TBOR + LEV groups. Under transmission electron microscopy, the corneas in the TBOR and TBOR + LEV groups were intact, whereas in the LEV and control groups, double-walled structures corresponding to S. aureus were found in the superficial stromal layer. Conclusions: TBOR demonstrated in vivo antibacterial efficacy against S. aureus. Translational Relevance: This study found in vivo antibacterial efficacy of toluidine blue-mediated photodynamic therapy on rabbit experimental bacterial keratitis, thus providing an additional new option for clinical treatment of bacterial keratitis.