TRPV4 regulates migration and tube formation of human retinal capillary endothelial cells.

BACKGROUND: Ca2+ entry plays an important role in modulating endothelial cell migration and tube formation. Transient receptor potential cation channel subfamily V member 4 (TRPV4) is a Ca2+-permeable channel that is widely expressed in endothelial cells. It has been reported that TRPV4 is expressed in HRCECs and regulates Ca2+ entry. However, the function of TRPV4 in human retinal capillary endothelial cells (HRCECs) remains unknown. METHODS: In this study we used western blot and immunostaining assay to verify TRPV4 expression in HRCECs. And then we pretreated HRCECs with HC067047 and transfected with specific shRNA of TRPV4. The functional presence of TrpV4 was determined by using fluorescence, migration and tube formation assay in TrpV4 knockdown cells or control cells. RESULTS: Using western blot and immunostaining, we confirmed TRPV4 expression in HRCECs. Moreover, inhibition of TRPV4 using the specific inhibitor HC067047 and the knockdown of TRPV4 with shRNA significantly suppressed tube formation and migration by HRCECs. CONCLUSIONS: TRPV4 is essential for HRCEC migration and tube formation, and maybe a potential therapeutic target for retinal vascular diseases.

Predictive visual field outcomes after optic chiasm decompressive surgery by retinal vessels parameters using optical coherence tomography angiography.

AIM: To evaluate the predictive value of superficial retinal capillary plexus (SRCP) and radial peripapillary capillary (RPC) for visual field recovery after optic cross decompression and compare them with peripapillary nerve fiber layer (pRNFL) and ganglion cell complex (GCC). METHODS: This prospective longitudinal observational study included patients with chiasmal compression due to sellar region mass scheduled for decompressive surgery. Generalized estimating equations were used to compare retinal vessel density and retinal layer thickness pre- and post-operatively and with healthy controls. Logistic regression models were used to assess the relationship between preoperative GCC, pRNFL, SRCP, and RPC parameters and visual field recovery after surgery. RESULTS: The study included 43 eyes of 24 patients and 48 eyes of 24 healthy controls. Preoperative RPC and SRCP vessel density and pRNFL and GCC thickness were lower than healthy controls and higher than postoperative values. The best predictive GCC and pRNFL models were based on the superior GCC [area under the curve (AUC)=0.866] and the tempo-inferior pRNFL (AUC=0.824), and the best predictive SRCP and RPC models were based on the nasal SRCP (AUC=0.718) and tempo-inferior RPC (AUC=0.825). There was no statistical difference in the predictive value of the superior GCC, tempo-inferior pRNFL, and tempo-inferior RPC (all P>0.05). CONCLUSION: Compression of the optic chiasm by tumors in the saddle area can reduce retinal thickness and blood perfusion. This reduction persists despite the recovery of the visual field after decompression surgery. GCC, pRNFL, and RPC can be used as sensitive predictors of visual field recovery after decompression surgery.

[Correlation between white-to-white diameter and ciliary sulcus diameter of high myopia eyes].

OBJECTIVE: To investigate the ciliary sulcus diameters in horizontal and vertical axes and to analyze the correlation between white-to-white diameter and horizontal ciliary sulcus diameter for high myopia eyes at different anterior chamber depths. METHODS: IOL-Master was used to measure white-to-white diameters and anterior chamber depths of 108 high myopia eyes and verified by digital caliper. Based on the depth of central anterior chamber not including corneal thickness, the testing eyes were divided into three groups: shallow AC group (3.0 mm < ACD ≤ 3.2 mm), medium AC group (3.2 mm < ACD ≤ 3.4 mm), and deep AC group (ACD > 3.4 mm). Furthermore, the horizontal and vertical ciliary sulcus diameters were checked by full-scale 50-Hz ultrasound biomicroscopy. The data of three AC groups were analyzed separately. The paired sample t-test was used to compare: white-to-white diameters and horizontal sulcus diameters, horizontal and vertical suclus diameters. The correlations between white-to-white diameters and horizontal sulcus diameters were assessed using linear regression analysis. The difference in the three groups of white-to-white diameters and ciliary sulcus diameters were compared by Student Newman Keuls test. RESULTS: The mean diameters and standard deviations of WTW diameters and horizontal ciliary sulcus diameters in the shallow AC group was, respectively, (11.46 ± 0.38) and (11.57 ± 0.32) mm(t = -2.849, P = 0.007). Linear regression analysis found a statistically significant correlation between white-to-white diameters and horizontal sulcus diameters with a coefficient of determinant of 0.797 (P = 0.000). The mean vertical suclus diameters was (12.27 ± 0.48) mm. The difference between horizontal and vertical suclus diameters was statistically significant (t = -15.522, P = 0.000) . The mean diameters and standard deviations of WTW diameters and horizontal ciliary sulcus diameters in the medium AC group was, respectively, (11.58 ± 0.31) and (11.77 ± 0.26) mm. (t = -4.523, P = 0.000). Linear regression analysis found a statistically significant correlation between white-to-white diameters and horizontal sulcus diameters with a coefficient of determinant of 0.639 (P = 0.000). The mean vertical suclus diameters was (12.52 ± 0.37) mm. The difference between horizontal and vertical suclus diameters was statistically significant (t = -13.987, P = 0.001) . The mean diameters and standard deviations of WTW diameters and horizontal ciliary sulcus diameters in the medium AC group was, respectively, (11.68 ± 0.22) and (11.91 ± 0.23) mm (t = -5.157, P = 0.000). Linear regression did not reveal horizontal sulcus diameters were a significant predictor of white-to-white diameters with a coefficient of determinant of 0.253 (P = 0.126). The mean vertical suclus diameters was (12.69 ± 0.36) mm. The difference between horizontal and vertical suclus diameters was statistically significant (t = -18.433, P = 0.000) . CONCLUSIONS: The white-to-white diameter measuring technique is not identical to the horizontal diameter of the ciliary sulcus and the potential difference between them is increased with deepening anterior chamber depth. Our preliminary study illustrated that the ciliary sculus is not circular and a vertical diameter is longer than horizontal one.

Outcomes of 1.8-3.0 mm incision phacoemulsification combined with trabeculectomy for primary angle-closure glaucoma with cataract.

AIM: To compare efficacy of coaxial microincisions (1.8 mm, 2.2 mm) and small incisions (3.0 mm) on phacoemulsification combined with trabeculectomy for primary angle-closure glaucoma (PACG) with cataract. METHODS: Ninety-six patients (96 eyes) with PACG and cataract were recruited and randomly divided into three groups between January 2015 and June 2017. Group A (3.0 mm incision), B (2.2 mm incision), and C (1.8 mm incision) comprised 30, 34 and 32 eyes respectively. All cases were treated with clear corneal incision phacoemulsification combined with trabeculectomy. Data including best corrected visual acuity (BCVA), corneal astigmatism, corneal endothelial cell counts (CECC), intraocular pressure (IOP), and complications were collected before the operation, and at postoperative 1d, 1 and 3mo. RESULTS: All the patients were successfully treated with surgery. The BCVA of groups B and C were significantly improved as compared to group A at postoperative 1d, 1 and 3mo (all P<0.05), but there was no difference between groups B and C at each time interval (all P>0.05). The corneal astigmatism of group A was statistically higher than that of group B (P=0.026); corneal astigmatism of group B was statistically higher than that of group C at postoperative 1d (P=0.006). The corneal astigmatism of group A at postoperative 3mo was significantly higher than that before operation (P=0.003). At postoperative 1 and 3mo, corneal astigmatism of groups B and C were significantly lower than that of group A (all P<0.05). The CECC in group B was significantly higher than that of group A (P=0.020), and CECC in group C was significantly higher than that of group B (P=0.034) at postoperative 1d. At postoperative 1 and 3mo, CECC of groups B and C were significantly higher than that of group A (all P<0.05). In each group, postoperative mean IOP at each time interval was significantly lower than preoperative IOP (all P<0.05). CONCLUSION: Coaxial microincision phacoemulsification combined with trabeculectomy for PACG with cataract has better curative efficacy in reducing postoperative corneal astigmatism and corneal endothelial cell injury than traditional small incision combined surgery, and the 1.8 mm microincision has better curative efficacy than 2.2 mm microincision in the early postoperative period.

Effects of astigmatic keratotomy combined with scleral tunnel incisions for the treatment of high astigmatism after penetrating keratoplasty.

The present study aimed to evaluate the efficacy, predictability and safety of astigmatic keratotomy (AK) combined with scleral tunnel incisions in the treatment of high astigmatism after penetrating keratoplasty (PKP). Paired AK combined with scleral tunnel incisions was performed at the steep astigmatic meridian in 8 eyes of 8 patients with high keratometric astigmatism [>5.0 diopters (D)] after PKP. Pre- and post-operative parameters, including uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), refraction and keratometric astigmatism were evaluated. The Alpins method for vector analysis was used to evaluate the changes in keratometric astigmatism. The results indicated a statistically significant reduction in the mean keratometric astigmatism from 8.16±3.02 D pre-operatively to 2.28±1.07 D at 3 months postoperatively. The mean UCVA improved from 0.95±0.24 logarithm of the minimum angle of resolution (logMAR) pre-operatively to 0.61±0.17 logMAR at 3 months postoperatively (P<0.05). The mean BCVA improved from 0.41±0.18 logMAR pre-operatively to 0.26±0.12 logMAR at 3 months postoperatively (P>0.05). Between 3 and 6 months after the surgery, the keratometric astigmatism remained stable. Alpins vector analysis demonstrated the relative predictability of this combined surgical treatment. The surgically induced astigmatism was significantly correlated with the target induced astigmatism (r=0.76, P<0.05). None of the patients had any severe complications. The present study indicated that AK combined with scleral tunnel incisions is an effective, relatively predictable and safe treatment for high astigmatism after PKP.

[Clinical research of intraoperative floppy iris syndrome during operation].

Intraoperative floppy iris syndrome (IFIS) has been recently identified as a new small pupil syndrome during phacoemulsification. This syndrome is characterized by three intraoperative features: a flaccid iris stroma that undulates and bellows in response to intraocular fluid currents; a propensity for the floppy iris stroma to prolapse toward the tip of phacoemulsification and side-port incisions despite proper wound construction; and progressive intraoperative pupil constriction despite standard preventive preoperative pharmacologic measures designed to prevent this. It is now mostly considered that IFIS is associated with the use of tamsolusin, a highly selective alpha-1A receptor antagonist for the treatment of benign prostatic hypertrophy. It is recommended that a careful history of the use of alpha-1 blocking agents be taken before cataract surgery to anticipate the occurrence of IFIS. A combination of strategies could decrease the complications of IFIS. These procedures include preoperative use of atropine, intracameral injection of dilute phenylephrine or epinephrine, the use of super-cohesive ophthalmic viscosurgical devices, lower phacoemulsification vacuum and aspiration settings and various iris hooks or pupil dilators.

Protective Effect of Rutin Against H2O2-Induced Oxidative Stress and Apoptosis in Human Lens Epithelial Cells.

PURPOSE: The aim of this study was to evaluate the effect of rutin on oxidative stress and apoptosis induced by H2O2 in human lens epithelial (HLE) cells and the associated mechanisms involved. METHODS: Cell viability was assessed by 4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide assay and cell apoptosis was determined by flow cytometry, TUNEL assay and DNA fragmentation assay after 24 h treatment of 100 µM H2O2 with or without rutin pretreatment at various concentrations. The level of reactive oxygen species (ROS) was examined using 2',7'-dichlorodihydrofluorescein diacetate by flow cytometry. The activity of superoxide dismutase (SOD) was measured by xanthinoxidase method and the contents of glutathione (GSH) and malondialdehyde (MDA) were quantified by enzyme-linked immunosorbent assay. The expression change of Bcl-2, Bax and caspase-3 at mRNA and protein levels were detected by real-time polymerized chain reaction (RT-PCR) and Western-blot analysis, respectively. Activation and translocation of nuclear factor-kappaB (NF-кB/p65) were examined by Western blot and immunocytochemistry. RESULTS: Rutin pretreatment protected HLE cells from H2O2-induced cell viability decrease and apoptosis. In addition, in the presence of rutin, H2O2-induced intracellular excessive ROS and MDA were attenuated, whereas intracellular SOD and GSH depletion were prevented. Moreover, rutin also inhibited the up-regulation of caspase-3 and Bax expression and rescued down-regulation of Bcl-2 expression. Lastly, rutin blocked the activation and translocation of NF-кB/p65 induced by H2O2. CONCLUSIONS: Our results demonstrated that rutin effectively protects HLE cells from H2O2-induced oxidative stress and apoptosis in a dose-dependent manner. The involved mechanisms may be related to the regulation of ROS production, the inhabitation of lipid peroxidation, the protection of intracellular antioxidant system and its modulation of Bcl-2/Bax family and NF-кB/p65 signaling pathway.

SMILE and Wavefront-Guided LASIK Out-Compete Other Refractive Surgeries in Ameliorating the Induction of High-Order Aberrations in Anterior Corneal Surface.

Purpose. To compare the change of anterior corneal higher-order aberrations (HOAs) after laser in situ keratomileusis (LASIK), wavefront-guided LASIK with iris registration (WF-LASIK), femtosecond laser-assisted laser in situ keratomileusis (FS-LASIK), and small incision lenticule extraction (SMILE). Methods. In a prospective study, 82 eyes underwent LASIK, 119 eyes underwent WF-LASIK, 88 eyes underwent FS-LASIK, and 170 eyes underwent SMILE surgery. HOAs were measured with Pentacam device preoperatively and 6 months after surgery. The aberrations were described as Zernike polynomials, and analysis focused on total HOAs, spherical aberration (SA), horizontal coma, and vertical coma over 6 mm diameter central corneal zone. Results. Six months postoperatively, all procedures result in increase of anterior corneal total HOAs and SA. There were no significant differences in the induced HOAs between LASIK and FS-LASIK, while SMILE induced fewer total HOAs and SA compared with LASIK and FS-LASIK. Similarly, WF-LASIK also induced less total HOAs than LASIK and FS-LASIK, but only fewer SA than FS-LASIK (P < 0.05). No significant difference could be detected in the induced total HOAs and SA between SMILE and WF-LASIK, whereas SMILE induced more horizontal coma and vertical coma compared with WF-LASIK (P < 0.05). Conclusion. FS-LASIK and LASIK induced comparable anterior corneal HOAs. Compared to LASIK and FS-LASIK, both SMILE and WF-LASIK showed advantages in inducing less total HOAs. In addition, SMILE also possesses better ability to reduce the induction of SA in comparison with LASIK and FS-LASIK. However, SMILE induced more horizontal coma and vertical coma compared with WF-LASIK, indicating that the centration of SMILE procedure is probably less precise than WF-LASIK.

Identification of Mutations in Myocilin and Beta-1,4-galactosyltransferase 3 Genes in a Chinese Family with Primary Open-angle Glaucoma.

BACKGROUND: Glaucoma is a major cause of irreversible blindness worldwide. There is evidence showing that a subset of the disease is genetically determined. In this study, we screened for mutations in chromosome 1q-linked open-angle glaucoma (GLC1A) in a Chinese family with primary open-angle glaucoma (POAG). METHODS: A total of 23 members from five generations of a family were enrolled and underwent thorough ophthalmologic examinations. In addition, 200 unrelated healthy Chinese controls were also recruited as normal control. GLC1A gene was amplified by polymerase chain reaction, and DNA sequencing was performed to screen for mutations. RESULTS: Six members were diagnosed as POAG, with severe clinical manifestations, and history of high intraocular pressures. The mean age of disease onset was 26.3 years. However, the others were asymptomatic. In six affected and three asymptomatic members, gene sequencing revealed a mutation c.C1456T in exon 3 of myocilin gene (MYOC). Furthermore, we also identified a novel mutation c.G322A in beta-1,4-galactosyltransferase 3 (B4GALT3) gene in all six affected and three asymptomatic members, which was not reported previously in POAG patients. The two newly identified variants were absent in other family members as well as controls. CONCLUSION: The mutations c.1456C < T (p.L486F) in MYOC and c.322G < A (p.V108I) in B4GALT3 are likely responsible for the pathogenesis of POAG in this family.

An Updated Meta-Analysis: Risk Conferred by Glutathione S-Transferases (GSTM1 and GSTT1) Polymorphisms to Age-Related Cataract.

Purpose. To study the effects of glutathione S-transferase M1 (GSTM1) and T1 (GSTT1) polymorphisms on age-related cataract (ARC). Methods. After a systematic literature search, all relevant studies evaluating the association between GSTs polymorphisms and ARC were included. Results. Fifteen studies on GSTM1 and nine studies on GSTT1 were included in this meta-analysis. In the pooled analysis, a significant association between null genotype of GSTT1 and ARC was found (OR = 1.229, 95% CI = 1.057-1.429, and P = 0.007). In subgroup analysis, the association between cortical cataract (CC) and GSTM1 null genotype was statistically significant (OR = 0.713, 95% CI = 0.598-0.850, and P < 0.001). In addition, GSTM1 null genotype was significantly associated with ARC causing risk to individuals working indoors and not individuals working outdoors. The association between GSTT1 null genotype and risk of ARC was statistically significant in Asians (OR = 1.442, 95% CI = 1.137-1.830, and P = 0.003) but not in Caucasians. Conclusions. GSTM1 positive genotype is associated with increased risk of CC and loses the protective role in persons who work outdoors. Considering the ethnic variation, GSTT1 null genotype is found to be associated with increased risk of ARC in Asians but not in Caucasians.

Ciliary sulcus diameters at different anterior chamber depths in highly myopic eyes.

PURPOSE: To determine the correlation between white-to-white (WTW) diameters and horizontal ciliary sulcus diameters at different anterior chamber depths (ACDs) in highly myopic eyes. SETTING: First Affiliated Hospital of Anhui Medical University, Hefei, China. DESIGN: Cross-sectional observational study. METHODS: Highly myopic eyes were divided into a shallow ACD group (2.8 to 3.2 mm), medium ACD group (3.2 to 3.4 mm), and deep ACD group (>3.4 mm). Ciliary sulcus diameters were measured by full-scale 50 MHz ultrasound biomicroscopy. RESULTS: In the shallow ACD group, linear regression showed that horizontal sulcus diameters were a significant predictor of WTW diameters (P<.001). The difference between them was greater than 0.50 mm in 13.1% of eyes. In the medium ACD group, linear regression showed horizontal sulcus diameters were a significant predictor of WTW diameters (P<.001). The difference between them was greater than 0.50 mm in 25.7% of eyes and greater than 1.0 mm in 2.8% of eyes. In the deep ACD group, linear regression did not show that horizontal sulcus diameters were a significant predictor of WTW diameters (P=.126). The difference between them was greater than 0.50 mm in 31.6% of eyes and greater than 1.0 mm in 5.3% of eyes. In all eyes, vertical diameters were greater than horizontal diameters. CONCLUSIONS: The WTW technique was inaccurate at predicting the horizontal diameter of the ciliary sulcus, and predictive errors increased with increasing ACDs. The posterior chambers were not perfectly circular, with vertical diameters being greater than horizontal diameters. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

A simple oligonucleotide biochip capable of rapidly detecting known mitochondrial DNA mutations in Chinese patients with Leber's hereditary optic neuropathy (LHON).

Leber's hereditary optic neuropathy (LHON) is a maternally transmitted disease. Clinically, no efficient assay protocols have been available. In this study, we aimed to develop an oligonucleotide biochip specialized for detection of known base substitution mutations in mitochondrial DNA causing LHON and to investigate frequencies of LHON relevant variants in Anhui region of China. Thirty-two pairs of oligonucleotide probes matched with the mutations potentially linked to LHON were covalently immobilized. Cy5-lablled targets were amplified from blood DNA samples by a multiplex PCR method. Two kinds of primary mutations 11778 G > A and 14484 T > C from six confirmed LHON patients were interrogated to validate this biochip format. Further, fourteen Chinese LHON pedigrees and twenty-five unrelated healthy individuals were investigated by the LHON biochip, direct sequencing and pyrosequencing, respectively. The biochip was found to be able efficiently to discriminate homoplasmic and heteroplasmic mtDNA mutations in LHON. Biochip analysis revealed that twelve of eighteen LHON symptomatic cases from the 14 Chinese pedigree harbored the mutations either 11778G > A, 14484T > C or 3460G > A, respectively, accounting for 66.7%. The mutation 11778G > A in these patients was homoplasmic and prevalent (55.5%, 10 of 18 cases). The mutations 3460G > A and 3394T > C were found to co-exist in one LHON case. The mutation 13708G > A appeared in one LHON pedigree. Smaller amount of sampling and reaction volume, easier target preparation, fast and high-throughput were the main advantages of the biochip over direct DNA sequencing and pyrosequencing. Our findings suggested that primary mutations of 11778G > A, 14484T > C or 3460G > A are main variants of mtDNA gene leading to LHON in China. The biochip would easily be implemented in clinical diagnosis.