Applications of metals and metal compounds in improving the sensitivity of microfluidic biosensors - a review.

The enhancement of detection sensitivity in microfluidic sensors has been a continuously explored field. Initially, many strategies for sensitivity improvement involved introducing enzyme cascade reactions, but enzyme-based reactions posed challenges in terms of cost, stability, and storage. Therefore, there is an urgent need to explore enzyme-free cascade amplification methods, which are crucial for expanding the application range and improving detection stability. Metal or metal compound nanomaterials have gained great attention in the exploitation of microfluidic sensors due to their ease of preparation, storage, and lower cost. The unique physical properties of metallic nanomaterials, including surface plasmon resonance, surface-enhanced Raman scattering, metal-enhanced fluorescence, and surface-enhanced infrared absorption, contribute significantly to enhancing detection capabilities. The metal-based catalytic nanomaterials, exemplified by Fe3O4 nanoparticles and metal-organic frameworks, are considered viable alternatives to biological enzymes due to their excellent performance. Herein, we provide a detailed overview of the applications of metals and metal compounds in improving the sensitivity of microfluidic biosensors. This review not only highlights the current developments but also critically analyzes the challenges encountered in this field. Furthermore, it outlines potential directions for future research, contributing to the ongoing development of microfluidic biosensors with improved detection sensitivity.

Quantization of Optic Disc Characteristics in Young Adults Based on Artificial Intelligence.

PURPOSE: This study aimed to automatically and quantitatively analyse the characteristics of the optic disc by applying artificial intelligence (AI) to fundus images. METHODS: A total of 1084 undergraduates were recruited in this cross-sectional study. The optic disc area, cup-to-disc ratio (C/D), optic disc tilt, and the area, width, and height of peripapillary atrophy (PPA) were automatically and quantitatively detected using AI. Based on axial length (AL), participants were divided into five groups: Group 1 (AL ≤ 23 mm); Group 2 (23 mm < AL≤ 24 mm); Group 3 (24 mm < AL≤ 25 mm); Group 4 (25 mm < AL< 26 mm) and Group 5 (AL ≥ 26 mm). Relationships between ocular parameters and optic disc characteristics were analysed. RESULT: A total of 999 undergraduates were included in the analysis. The prevalence of optic disc tilting and PPA were 47.1% and 92.5%, respectively, and increased with the severity of myopia. The mean optic disc area, PPA area, C/D, and optic disc tilt ratio were 1.97 ± 0.46 mm2, 0.84 ± 0.59 mm2, 0.18 ± 0.07, and 0.81 ± 0.08, respectively. In Group 5, the average optic disc area (1.84 ± 0.41 mm2) and optic disc tilt ratio (0.79 ± 0.08) were significantly smaller and the PPA area (1.12 ± 0.61 mm2) was significantly larger than those in the other groups. AL was negatively correlated with optic disc area and optic disc tilt ratio (r=-0.271, -0.219; both p < 0.001) and positively correlated with PPA area, width, and height (r = 0.421, 0.426, 0.345; all p < 0.01). A greater AL (ß = 0.284, p < 0.01) and a smaller optic disc tilt ratio (ß=-0.516, p < 0.01) were related to a larger PPA area. CONCLUSION: The characteristics of the optic disc can be feasibly and efficiently extracted using AI. The quantization of the optic disc might provide new indicators for clinicians to evaluate the degree of myopia.

Pseudomyopia as an independent risk factor for myopia onset: a prospective cohort study among school-aged children.

AIM: To investigate whether pseudomyopia is an independent risk factor for myopia onset based on a population-based cohort study. METHODS: Non-myopic children were recruited from schools in rural and urban settings of Shangdong province, China. Baseline examinations started in September 2020 and all participants were invited for a 6-month follow-up. Pseudomyopia was defined as spherical equivalent (SE) ≤-0.50 diopters (D) before cycloplegia and >-0.50D after cycloplegia. Myopia was defined as cycloplegic SE ≤-0.50D. RESULTS: A total of 2328 children (baseline age: 4-17 years) were included in the final analysis. During the 6-month follow-up, 21.1% (355/1680) pseudomyopic eyes developed myopia, and 3.8% (110/2879) non-myopic and non-pseudomyopic eyes developed myopia. After adjusting for multiple myopia risk factors, including baseline cycloplegic SE, near work and outdoor time, pseudomyopia was found to be an independent risk factor for myopia onset (relative risk=2.52, 95% CI 1.86 to 3.42). Additionally, pseudomyopic children with more myopic cycloplegic SE (p<0.001), smaller difference between cycloplegic and non-cycloplegic SE (DIFF, p<0.001), and higher binocular amplitude of accommodation (p<0.001) had higher risk of myopia development. CONCLUSION: This is an important longitudinal study to prove that pseudomyopia is an independent risk factor for myopia development among school-aged children.

Widespread Involvement of Acetylation in the Retinal Metabolism of Form-Deprivation Myopia in Guinea Pigs.

Myopia has become the major cause of visual impairment worldwide. Although the pathogenesis of myopia remains controversial, proteomic studies suggest that dysregulation of retinal metabolism is potentially involved in the pathology of myopia. Lysine acetylation of proteins plays a key role in regulating cellular metabolism, but little is known about its role in the form-deprived myopic retina. Hence, a comprehensive analysis of proteomic and acetylomic changes in the retinas of guinea pigs with form-deprivation myopia was performed. In total, 85 significantly differential proteins and 314 significantly differentially acetylated proteins were identified. Notably, the differentially acetylated proteins were markedly enriched in metabolic pathways such as glycolysis/gluconeogenesis, the pentose phosphate pathway, retinol metabolism, and the HIF-1 signaling pathway. HK2, HKDC1, PKM, LDH, GAPDH, and ENO1 were the key enzymes in these metabolic pathways with decreased acetylation levels in the form-deprivation myopia group. Altered lysine acetylation of key enzymes in the form-deprived myopic retina might affect the dynamic balance of metabolism in the retinal microenvironment by altering their activity. In conclusion, as the first report on the myopic retinal acetylome, this study provides a reliable basis for further studies on myopic retinal acetylation.

Therapeutic applications of contact lens-based drug delivery systems in ophthalmic diseases.

Traditional ophthalmic drugs, such as eye drops, gels and ointments, are accompanied by many problems, including low bioavailability and potential drug side effects. Innovative ophthalmic drug delivery systems have been proposed to overcome the limitations associated with traditional formulations. Recently, contact lens-based drug delivery systems have gained popularity owing to their advantages of sustained drug delivery, prolonged drug retention, improved bioavailability, and few drug side effects. Various methods have been successfully applied to drug-loaded contact lenses and prolonged the drug release time, such as chemical crosslinking, material embedding, molecular imprinting, colloidal nanoparticles, vitamin E modification, drug polymer film/coating, ion ligand polymerization systems, and supercritical fluid technology. Contact lens-based drug delivery systems play an important role in the treatment of multifarious ophthalmic diseases. This review discusses the latest developments in drug-loaded contact lenses for the treatment of ophthalmic diseases, including preparation methods, application in ophthalmic diseases and future prospects.

Application of Artificial Intelligence to Quantitative Assessment of Fundus Tessellated Density in Young Adults with Different Refractions.

INTRODUCTION: The aim of this study was to quantitatively assess fundus tessellated density (FTD) and associated factors by artificial intelligence (AI) in young adults. METHODS: A total of 1,084 undergraduates (age, 17-23 years old) were enrolled in November 2021. The students were divided into three groups according to axial length (AL): group 1 (AL <24.0 mm, n = 155), group 2 (24 mm ≤ AL <26 mm, n = 578), and group 3 (AL ≥26 mm, n = 269). FTD was calculated by extracting the fundus tessellations as the regions of interest (circle 1, diameter of 3.0 mm; circle 2, diameter of 6.0 mm) and then calculating the average exposed choroid area per unit area of fundus. RESULTS: Among 1,084 students, 1,002 (92.5%) students' FTDs were extracted. The mean FTD was 0.06 ± 0.06 (range, 0-0.40). In multivariate analysis, FTD was significantly associated with male sex, longer AL, thinner subfoveal choroid thickness (SFCT), increased choriocapillaris vessel density (VD), and decreased deeper choroidal VD (all p < 0.05). In circle 1 (diameter of 3.0 mm) and circle 2 (diameter of 6.0 mm), analysis of variance showed that the FTD of the nasal region (p < 0.05) was significantly larger than that of the superior, inferior, and temporal regions. CONCLUSION: AI-based imaging processing could improve the accuracy of fundus tessellation diagnosis. FTD was significantly associated with a longer AL, thinner SFCT, increased choriocapillaris VD, and decreased deeper choroidal VD.

Electroacupuncture Improves Choroidal Blood Flow to Inhibit the Development of Lens-Induced Myopia in Guinea Pigs.

Introduction: The purpose of this paper was to study the effect of electroacupuncture (EA) on choroidal blood flow (ChBF) in a guinea pig model of lens-induced myopia (LIM). Methods: Guinea pigs were randomly divided into 4 groups: normal control (NC) group, LIM group, LIM + electroacupuncture (LIM + EA) group, and LIM + sham acupoint (LIM + sham) group. Right eyes were covered with a -6D lens to induce myopia. Meanwhile, LIM + EA group and LIM + sham group were treated with EA at acupoints Hegu (LI4) and Taiyang (EX-HN5) and sham points. Refraction, axial length (AL), choroidal thickness (ChT), vessel density of choriocapillaris (CC) and choroidal layer, and scleral collagen fiber were measured. Besides, hypoxia-inducible factor-1α (HIF-1α), matrix metalloprotein-2 (MMP-2), and tissue inhibitor metalloprotease-2 (TIMP-2) expression in sclera were detected. Results: Refraction and AL were significantly decreased and ChT and vessel density of CC were significantly increased in LIM + EA group at 2 weeks and 4 weeks (all P < 0.05) compared with LIM group. However, no significant difference of vessel density of choroidal layer was observed between LIM and LIM + EA group at 2 weeks and 4 weeks. Scleral collagen fibrils diameters were significantly increased in LIM + EA group at 4 weeks (P < 0.001) compared with LIM group. At the end of experiment, the mRNA and protein expression of HIF-1α and MMP-2 were significantly decreased (all P < 0.05) and those of TIMP-2 were increased in LIM + EA, compared with LIM. However, there were no significant differences between LIM and LIM + sham group. Conclusions: EA can improve the vessel density of choroid and then possibly improve scleral hypoxia, which may inhibit the growth of the AL in myopia guinea pig.

In vivo analysis of ciliary muscle in myopic Chinese young adults using ArcScan Insight® 100.

OBJECTIVE: To analyse the morphological characteristics of the ciliary muscle (CM) and to explore its relationship with different ocular biometric parameters in myopic young Chinese adults. METHODS: This observational, cross-sectional study included 50 right eyes from 50 myopic adults. The CM area (CMA), CM thickness (CMT) and CM length (CML) were measured using the ArcScan Insight® 100. CMT was determined at three points: 1.0 mm (CMT-1), 2.0 mm (CMT-2) and 3.0 mm (CMT-3) posterior to the scleral spur. CML was measured on the scleral (CMLs) and vitreous (CMLv) aspects. The spherical equivalent refraction (SER), axial length (AL) and subfoveal choroidal thickness (SFCT) were examined to determine their associations with CM parameters (CMA, CML and CMT). RESULTS: The mean SER and AL were -4.39 ± 2.29 D and 25.61 ± 1.15 mm, respectively. Compared with the nasal CMA, CML and CMT (CMT-1, CMT-2 and CMT-3) findings, the temporal CM parameters (CMA, CMLs, CMLv, CMT-1, CMT-2 and CMT-3) were found to be significantly thicker (all p < 0.001, except CMLv and CMT-1; p < 0.01). The nasal CMA was associated with the average corneal curvature (r = 0.30, p = 0.03) and SER (r = -0.30, p = 0.04). Nasal and temporal CMT-2 were negatively correlated with SER (r = -0.33 and -0.32, respectively, both p < 0.05). There was no correlation between CM parameters (except nasal CMLs, r = 0.31, p = 0.03) and SFCT, or between CM parameters and either the AL or anterior chamber depth (all p > 0.05). CONCLUSION: These results suggest that there is temporal versus nasal asymmetry of the CM. CMA, CMT or CML did not vary with axial growth of the eye. The CM is not simply stretched as the eye elongates in myopic young adults.

Rabbit-derived human Thelazia callipaeda infection: a case report.

In November 2021, a 19-year-old medical student was diagnosed with a rabbit-derived Thelazia callipaeda infection in her left eye. She visited her doctor because she removed the white object by herself with a cotton swab 2 days earlier. She explained to the doctor that she rubbed her eyes without washing her hands when rabbit hair got into her eyes during the animal experiment class at school 2 weeks earlier. The other 2 worms were then taken out with ophthalmic forceps and were placed into physiological saline. The worms were sent to the Shandong Academy of Eye Disease Prevention and Therapy for examination. After identification, the worms were regarded as the female T. callipaeda. Ofloxacin eye drops and peranapulin eye drops were administered to control inflammation. No abnormality was found in the follow-up 2 weeks and 1 month later. This report highlights that it is very important to strengthen the health and safety education of medical students during animal experiment classes.

Epigallocatechin gallate enhances human lens epithelial cell survival after UVB irradiation via the mitochondrial signaling pathway.

The aim of the present study was to explore the mechanism underlying the ultraviolet B (UVB) irradiation­induced apoptosis of human lens epithelial cells (HLECs), and to investigate the protective effect of epigallocatechin gallate (EGCG) against the UVB­induced apoptosis of HLECs. HLECs were exposed to different concentrations of EGCG plus UVB (30 mJ/cm2). Cell viability was determined using the MTT assay. Furthermore, mitochondrial membrane potential (Δψm) and apoptosis were assessed by flow cytometry with JC­1 and Annexin V/PI staining, respectively. Moreover, the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH­Px), as well as the levels of GSH, hydrogen peroxide (H2O2) and hydroxyl free radicals were determined using biochemical assay techniques. Reverse transcription­quantitative PCR and western blotting were used to detect the mRNA and protein expression levels of Bcl­2, Bax, cytochrome c, caspase­9 and caspase­3, respectively. The results revealed that UVB irradiation reduced the Δψm of HLECs and induced apoptosis. Notably, EGCG significantly attenuated the generation of H2O2 and hydroxyl free radicals caused by UVB irradiation in HLECs, and significantly increased CAT, SOD and GSH­Px activities, however, the GSH levels were not significantly increased. EGCG also reduced UVB­stimulated Bax, cytochrome c, caspase­9 and caspase­3 expression, and elevated Bcl­2 expression, suggesting that EGCG may possess free radical­scavenging properties, thus increasing cell viability. In conclusion, EGCG may be able to protect against UVB­induced HLECs apoptosis through the mitochondria­mediated apoptotic signaling pathway, indicating its potential application in clinical practice.

Stress Suppresses Systemic Th17/Treg Imbalance in Rats with Experimental Autoimmune Uveitis.

PURPOSE: To explore the effect of chronic unpredictable mild stress (CUMS) on the immune response involved in rats with experimental autoimmune uveitis (EAU). METHODS: Lewis rats were randomly divided into control, EAU, CUMS, and EAU+CUMS groups and received relevant treatments. On days 7, 11, 14, 21 and 28, frequencies of Th17 and Treg cells and the related cytokines were analyzed. RESULTS: The intraocular inflammation of EAU rats peaked between days 11 and 13, while the severity of inflammation of the rats in EAU+CUMS group fluctuated between 11 and 15 days. Both frequencies of Th17, Treg cells and the related cytokines exhibited a significant difference between the two groups on days 11 and 14. CONCLUSION: CUMS may protect against the possible harmful effects of immune disorder in rats with EAU through suppressing the immune disorder of T lymphocyte and the related cytokine responses.

Refractive lens power and lens thickness in children (6-16 years old).

To examine the refractive lens power (RLP) and lens thickness and their associated factors in children from North-Western China. Children from two schools (primary school and junior high school) in the North-Western Chinese province of Qinghai underwent a comprehensive ophthalmic examination including biometry and cycloplegic refractometry. The RLP was calculated using Bennett's equation. The study included 596 (77.9%) individuals (mean age: 11.0 ± 2.8 years; range: 6-16 years) with a mean axial length of 23.65 ± 1.24 mm (range: 20.02-27.96 mm). Mean lens thickness was 3.30 ± 0.16 mm (range: 2.85-3.99 mm) and mean RLP was 24.85 ± 1.98D (range: 19.40-32.97). In univariate analysis, girls as compared to boys had a significantly thicker lens and greater RLP, shorter axial length, smaller corneal curvature radius and shorter corneal curvature radius (all P < 0.001). Both sexes did not differ significantly in refractive error (P = 0.11) and corneal thickness (P = 0.16). RLP was positively associated with refractive error (correlation coefficient r = 0.33; P < 0.001) and lens thickness (r = 0.62; P < 0.001) and negatively with axial length (r = - 0.70; P < 0.001). In univariate analysis, RLP decreased significantly with older age in the age group from age 6-13, while it plateaued thereafter, with no significant difference between boys and girls. In multivariate regression analysis, a higher RLP was associated with younger age (P < 0.001; standard regression coefficient ß = - 0.07), female sex (P < 0.001; ß = - 0.08), shorter axial length (P < 0.001; ß = - 0.48) and higher lens thickness (P < 0.001; ß = 0.42). In Chinese children, RLP with a mean of 24.85 ± 1.98D decreases with older age, male sex, longer axial length, and thinner lens thickness. Changes in RLP and axial length elongation are important players in the emmetropization and myopization.

Prevalence of Refractive Error and Visual Acuity Among School Children in the Plateau Region of Qinghai, China.

PURPOSE: This study aimed to investigate the prevalence of refractive error and visual acuity among school children in the plateau region of Qinghai, China. METHODS: The school-based, cross-sectional study was performed in Menyuan, Qinghai, China. Three kindergartens and three primary schools were randomly enrolled from both rural areas and county towns. The participants had undergone ophthalmic examinations of the intraocular pressure (IOP), uncorrected visual acuity (UCVA), presenting visual acuity (PVA) and best corrected visual acuity (BCVA), cycloplegic refraction, and axial length (AL). Regression analysis was applied to investigate the potential risk factors affecting the prevalence of various refractive errors. RESULTS: A total of 3770 children were invited to participate, and 3524 (93.5%) had undergone examination. Among the 3524 children (51.8% boys) with a mean age of 8.3±2.3 years, 1049 (29.8%) had myopia, 30 (0.9%) had high myopia, 1692 (48.0%) had mild hyperopia, 152 (4.3%) had medium to marked hyperopia and 925 (26.2%) had astigmatism. The mean SER was -0.16±1.86 D and decreased with age. The AL increased with age from 21.80±0.59 mm at 4-years to 23.53±1.05 mm at 12-years. The myopia prevalence increased with age from 2.0% at 4 years to 62.8% at 12-years. Myopia was associated with increasing age, county town habitation and girls. Among the 723 participants with PVA 20/40 or worse in one eye, 564 (78.0%) were due to uncorrected refractive error, and 83 (22.0%) were due to undercorrected refractive error. Among the 1049 children with myopia, only 254 wore glasses, and 151 children with PVA had a worse BCVA and did not have accurate spectacles. CONCLUSION: The prevalence of myopia is very high among school children in Menyuan. Only 24.2% of myopic children wore glasses, and 59.4% of children did not have accurate spectacles. Strategies to improve access to eye care and affordable glasses are needed.

Effects of Orthokeratology on Axial Length Elongation in Anisometropes.

INTRODUCTION: This study aimed to investigate the therapeutic effects of overnight orthokeratology (OK) lenses on anisometropes. METHODS: We enrolled 178 anisometropes from August 2015 to August 2017. We then divided these patients into 2 parts depending on them wearing either monocular or binocular OK lenses. In part one, 47 monocular myopic subjects (25 males and 22 females) were treated with OK lenses in the myopic eyes only. We also labeled the myopic eyes as the OK group and the contralateral nonmyopic eyes as the control group. The initial average wearing age of the subjects was 12.35 ± 2.37 years (8-16 years). The mean follow-up duration was 15.43 ± 4.88 months (7-25 months). The average spherical equivalent refraction (SER) was -2.31 ± 1.16 diopter (D) in the OK group and 0.15 ± 0.49 D in the control group (p < 0.001). In part 2, 131 binocular myopic anisometropes (56 males and 75 females) were involved in the study. The eyes with more severe myopia were assigned to the G group and the contralateral eyes to the L group. The initial average wearing age of the subjects was 12.92 ± 2.60 years ( 8-16 years). The mean follow-up duration was 17.83 ± 5.02 months (7-26 months). The average SER was -4.79 ± 1.90 D in the G group and -3.14 ± 1.88 D in the L group (p < 0.001). We calculated the axial length (AL) difference and AL elongation as our primary outcome measures. RESULTS: In part one, the AL elongation in the OK group (0.21 ± 0.09 mm) was significantly lower than that in the control group (0.70 ± 0.17 mm) at 24 months (p < 0.001). Meanwhile, the AL difference exhibited a decrease of 0.50 ± 0.29 mm from a baseline of 1.08 ± 0.35 to 0.58 ± 0.25 mm at 24 months (F = 24.539, p < 0.001). In part 2, the AL had increased by 0.17 ± 0.13 mm in the G group and 0.24 ± 0.18 mm in the L group after 24-month follow-up, respectively (p < 0.001). While the AL difference decreased from 0.55 ± 0.11 mm at the baseline, to 0.48 ± 0.08 mm at 24 months, eliciting a decrement in AL difference of 0.07 ± 0.09 mm (F = 3.884, p = 0.030). CONCLUSIONS: OK lenses can slow down AL growth in anisometropes and has a greater effect on reducing AL elongation in the more severely affected myopic eyes of anisometropic patients.

Choroidal changes in lens-induced myopia in guinea pigs.

INTRODUCTION: This study aimed to determine the role of the choroid in lens-induced myopia (LIM) in guinea pigs. METHODS: Guinea pigs were randomly divided into two groups: a normal control (NC) group and a LIM group. Refraction and axial length (AL) were measured by streak retinoscopy and A-scan ultrasonography. The choroidal thickness (ChT), vessel density of the choriocapillaris (VDCC) and vessel density of the choroidal layer (VDCL) were assessed by Spectral-domain Optical Coherence Tomography Angiography (SD-OCT). In addition, the choroidal expression of nitric oxide synthase (NOS) enzymes at the mRNA and protein levels was analyzed by real-time fluorescence quantitative PCR, enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry. RESULTS: In the LIM group, refraction and AL were increased significantly compared with those in the NC group at 2 weeks (refraction: LIM vs. NC, -4.23 ± 0.43 D vs. 2.20 ± 0.48 D; AL: LIM vs. NC, 8.36 ± 0.05 mm vs. 8.22 ± 0.03 mm) and 4 weeks (refraction: LIM vs. NC, -5.88 ± 0.49 D vs. 1.63 ± 0.41 D; AL: 8.57 ± 0.06 mm vs. 8.40 ± 0.04 mm). The ChT and VDCC were decreased significantly compared with those in the NC group at 2 weeks (ChT: LIM vs. NC, 60.92 ± 8.15 µm vs. 79.11 ± 7.47 µm; VDCC: LIM vs. NC, 23.43 ± 3.85% vs. 28.74 ± 4.11%) and 4 weeks (ChT: LIM vs. NC, 48.43 ± 6.85 µm vs. 76.38 ± 7.84 µm; VDCC: LIM vs. NC, 21.29 ± 2.17% vs. 27.64 ± 2.91%). The VDCL was also decreased compared with that in the NC group at 2 weeks and 4 weeks (NC vs. LIM, 24.87 ± 5.16% vs. 22.45 ± 3.26%; 23.37 ± 5.85% vs. 21.39 ± 2.62%; all P > 0.05). Moreover, the ChT was positively correlated with the VDCC and VDCL. The mRNA and protein expression of NOS enzymes (eNOS and nNOS) was increased. CONCLUSIONS: During the development of myopia, the ChT, VDCC and VDCL were decreased, while NOS expression in the choroid was increased. The expression of NOS was negatively correlated with the ChT, VDCC and VDCL. NO may play an important role in regulating the choroid during myopia development.

Epigallocatechin gallate protects the human lens epithelial cell survival against UVB irradiation through AIF/endo G signalling pathways in vitro.

OBJECTIVE: Oxidative stress has been recognised as an important mediator of apoptosis in lens epithelial cells. It also plays an important role in the pathogenesis of cataracts. It is reported that (-)-Epigallocatechin gallate (EGCG), the most abundant component in green tea, exhibits potent antioxidant activity against oxidative stress. This study aimed to investigate the protective effect of EGCG against Ultraviolet B (UVB) induced apoptotic death and the underlying mechanism in human lens epithelial cells (HLECs). METHODS: HLECs were exposed to various concentrations of EGCG under UVB (30 mJ/cm2), and cell viability was monitored by the MTT assay. Next, mitochondrial membrane potential (Δψm), reactive oxygen species (ROS) and apoptosis were detected by flow cytometry. Meanwhile, the total antioxigenic capacity (T-AOC) was determined by enzyme standard instrument, and the expression of apoptosis inducing factor (AIF) and endonuclease G (Endo G) was measured by quantitative PCR (Q-PCR) and western blotting, respectively. Moreover, the localisation of AIF and Endo G within cells was further detected by confocal optical microscopy. RESULTS: The results indicated that EGCG could enhance the cell viability and protect against cell apoptosis caused by UVB irradiation in HLECs. EGCG could also decrease the UVB-induced generation of ROS and collapse of Δψm, increase the T-AOC level. In addition, EGCG could also inhibit the UVB-stimulated increase of AIF and Endo G expression at mRNA and protein levels and ameliorate the UVB-induced mitochondria-nuclear translocation of AIF and Endo G. CONCLUSIONS: UVB irradiation could damage HLECs viability, while EGCG exhibits antioxidant effect and inhibits UVB-induced apoptosis in HLECs through AIF/Endo G signalling pathways. Our findings reveal the underlying mechanism of EGCG against UVB-induced oxidative stress in HLECs.

Chlorogenic acid attenuates hydrogen peroxide­induced oxidative stress in lens epithelial cells.

Oxidative stress has an important role in the degradation, oxidation, cross­linking and aggregation of lens proteins, and can trigger lens epithelial cell apoptosis. To investigate the protective effect of chlorogenic acid (CGA) against hydrogen peroxide (H2O2)­induced oxidative stress, human lens epithelial cells (hLECs) were exposed to various concentrations of H2O2 in the presence and absence of CGA. Using MTT assay, reverse transcription­quantitative polymerase chain reaction (RT­qPCR) and ELISA techniques, cell viability, and protein/mRNA levels of BCL2 apoptosis regulator (Bcl­2) and BCL2 associated X apoptosis regulator (Bax) were investigated. Additionally, the levels of intracellular reactive oxygen species (ROS) and apoptosis within cells were measured using flow cytometry to determine the protective effect of CGA on H2O2­induced oxidative stress. Furthermore, the protective effect of CGA on H2O2­induced apoptosis was also examined using rabbit lenses ex vivo. The results indicated that CGA reduced H2O2­induced cytotoxicity in a dose­dependent manner. Flow cytometry analysis demonstrated that simultaneous exposure of hLECs to H2O2 and CGA significantly decreased apoptosis and the levels of ROS. RT­qPCR analysis revealed a decrease in Bcl­2 and an increase in Bax in hLECs following exposure to H2O2 for 24 h, regardless of CGA presence. Furthermore, ELISA results indicate that CGA increased Bcl­2 expression and decreased Bax expression following treatment with H2O2 for 24 h and the Bax/Bcl-2 ratio was significantly decreased by CGA treatment. Lens organ culture experiments indicated a dose­dependent decrease in H2O2­induced lens opacity following CGA treatment. These results suggest that CGA suppresses hLECs apoptosis and prevents lens opacity induced by H2O2 via Bax/Bcl­2 signaling pathway. CGA may provide effective defenses against oxidative stress and, thus, haσ potential as treatment for a variety of diseases in clinical practice.

Imbalance Between Th17 Cells and Regulatory T Cells During Monophasic Experimental Autoimmune Uveitis.

The aim of this study is to explore the dynamic changes in IL-17-expressing T cells (Th17)/Treg expression in monophasic experimental autoimmune uveitis (mEAU). mEAU was induced in Lewis rats with IRBP1177-1191 peptide and evaluated clinically and pathologically on days 9, 13, 18, 23, 28, 35, and 48. Lymphocytes isolated from inguinal lymph nodes were subjected to flow cytometry to analyze the frequency of Th17/Treg cells. The levels of cytokines (IL-17, IL-6, IL-10, transforming growth factor (TGF)-ß) in serum were detected by enzyme-linked immunosorbent assay (ELISA). Real-time quantitative PCR (RT-PCR) was used for measuring the levels of IL-17, IL-6, TGF-ß, and Foxp3. Clinical and histopathologic assessment showed that mEAU began on day 9, peaked on day 13, and decreased to normal on day 18. The frequency of Th17 cells increased obviously on day 9, peaking on day 13, while the frequency of Treg cells increased on day 13, peaked on day 18, and remained at a high level until day 48. In the serum, the levels of IL-17 and IL-6 peaked on day 9 and gradually decreased to normal on day 28. The level of TGF-ß increased on day 9, peaked on day 13, and decreased to normal on day 35. Meanwhile, the level of IL-10 increased on day 9 and stayed at a high level until day 48. Additionally, the above results were further confirmed by RT-PCR. The imbalance between Th17 and Treg cells contributes to the onset and progression of mEAU, and a compartmental imbalance of Treg over Th17 exists in the recovery phase of mEAU.

Pearl cyst treatment by surgical excision.

PURPOSE: Epithelial implantation cysts are rare complications of traumatic injury or incisional surgery resulting from transfer of surface epithelial cells from the cornea, conjunctiva, or periocular skin into the anterior chamber. They can either appear as solid, pearl white, or clear in the case of serous cysts. This study describes the clinical presentation, histopathological features, and surgical outcomes of a pearl cyst. CASE REPORT: A 19-year-old male subject first presented with corneal injury from a stone. After primary wound management, an eyelash was removed from the anterior chamber. The corneal wound was not sutured. Two months later, the patient was rehospitalized with a white mass in the anterior chamber. The mass was excised. Postoperative visual acuity was normal. Histological examination revealed an implantation cyst on the surface of the iris extensively covered with epithelial cells four to five layers thick in most areas. The patient has been followed up for 6 months without complications. CONCLUSIONS: The proliferation of traumatically implanted epithelial cells into the anterior chamber may lead to the development of a pearl cyst. The positive outcome in this case was consistent with the surgical treatment of choice evolving in the literature.

Preparation and evaluation of sinomenine hydrochloride in situ gel for uveitis treatment.

PURPOSE: The aim of the present study was to develop and optimize sinomenine hydrochloride (SIN) in situ gel for uveitis treatment. METHODS: Carbopol 940 was used as the gelling agent in combination with hydroxypropyl methylcellulose (HPMC), which acts as a viscosity enhancer. The formulations were prepared using various concentrations of Carbopol 940 and HPMC. The prepared in situ gels were evaluated for gellation, drug release, ocular irritation, elimination time and pharmacokinetic studies. Furthermore, the effect of SIN on the development of experimental autoimmune anterior uveitis (EAAU) was assessed. RESULTS: The optimum concentration of Carbopol was 0.1% (w/v), and that for HPMC was 0.4% (w/v). Which showed a significant enhancement in gel strength in the physiological condition while free flowing at non-physiological condition. Optimum formula F(2-3) consisting of 0.5% SIN was prepared and kept as gel group, and 0.5% SIN solution was prepared and kept as control group. Gel group provided sustained release of the drug over a period of 480 min. No evidence of overt toxicity and irritation was observed in any study. The elimination time of control group and gel group was completed within 10 min and 25 min, respectively. The area under the aqueous humor concentration vs. time curve (AUC(0-t)) and maximum concentration (C(max)) values of gel group was 2.70-fold and 1.79-fold higher than that of control group. Additionally, clinical examination showed that SIN suppressed inflammation in EAAU. CONCLUSIONS: These results support the potential applications of SIN in situ gel for uveitis treatment.