Zhejiang Provincial Key Laboratory of Ophthalmology

Hangzhou, China

Zhejiang Provincial Key Laboratory of Ophthalmology

Hangzhou, China
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Yu Y.,Zhejiang University | Yu Y.,Zhejiang Provincial Key Laboratory of Ophthalmology | Wu M.,Zhejiang University | Wu M.,Zhejiang Provincial Key Laboratory of Ophthalmology | And 6 more authors.
Scientific Reports | Year: 2016

Autosomal dominant congenital cataracts (ADCC) are clinically and genetically heterogeneous diseases. The present study recruited two Chinese families with bilateral nuclear cataract or zonular pulverulent phenotype. Direct sequencing of candidate genes identified two novel missense mutations of Cx50, Cx50P59A (c.175C > G) and Cx50R76H (c.227G > A), both co-segregated well with all affected individuals. Bioinformatics analysis predicted deleterious for both mutations. Functional and cellular behaviors of wild type and mutant Cx50 examined by stably transfecting recombinant systems revealed similar protein expression levels. Protein distribution pattern by fluorescence microscopy showed that Cx50R76H localized at appositional membranes forming gap junctions with enormous cytoplasmic protein accumulation, whereas the Cx50P59A mutation was found inefficient at forming detectable plaques. Cell growth test by MTT assay showed that induction of Cx50P59A decreased cell viability. Our study constitutes the first report that the Cx50P59A and Cx50R76H mutations are associated with ADCC and expands the mutation spectrum of Cx50 in association with congenital cataracts. The genetic, cellular, and functional data suggest that the altered intercellular communication governed by mutated Cx50 proteins may act as the molecular mechanism underlying ADCC, which further confirms the role of Cx50 in the maintenance of human lens transparency.


Luo Y.,Zhejiang University | Luo Y.,Zhejiang Provincial Key Laboratory of Ophthalmology | Liu S.,Zhejiang University | Liu S.,Zhejiang Provincial Key Laboratory of Ophthalmology | And 2 more authors.
Molecular Therapy - Nucleic Acids | Year: 2017

m-miR-184 (mutant miR-184, r.57c > u) appears in familial hereditary ocular diseases, including keratoconus, cataracts, EDICT (endothelial dystrophy, iris hypoplasia, congenital cataract, and stromal thinning) syndrome, severe keratoconus, and non-ectatic corneal thinning. The biological function of m-miR-184 in these ocular diseases remains unclear. With the emergence of high-throughput sequencing, it is now possible to discover many different biological components simultaneously. Using two different RNA libraries, we sequenced the complete transcriptome of HLE cells treated with miR-184, m-miR-184, and a negative control. Data were integrated in an effort to identify any novel gene affected by m-miR-184. Notably, we concluded that ALDH5A1 and GABRA3 were disordered by m-miR-184, which might lead to ocular disease. Moreover, circRNA (circular RNA) expression was highy random across miR-184, m-miR-184, and negative control treatment groups. The sequences of the circRNAs did reveal a particularly high level of ALU sequences. In summary, we provide a new avenue for understanding the role of m-miR-184 in ocular diseases. © 2017 The Author(s)


Sun C.-B.,Zhejiang University | Sun C.-B.,Zhejiang Provincial Key Laboratory of Ophthalmology | Teng W.-Q.,Zhejiang University | Cui J.-T.,Zhejiang University | And 3 more authors.
Colloids and Surfaces B: Biointerfaces | Year: 2014

Migration and epithelial-mesenchymal transition (EMT) of lens epithelial cells (LECs) are main causes of central posterior capsule opacification after cataract extraction combined with intraocular lens (IOL) implantation. In this study, commercially available hydrophobic acrylic IOLs were first pretreated with atmospheric pressure glow discharge plasma to produce plenty of negatively charged chemical groups onto IOL surface, then polyethylenimine was deposited onto IOL surfaces as a precursor monolayer, and then anti-TGF-β2 (anti-T) antibody and poly-l-lysine were sequentially deposited onto IOL surface for four cycles followed by another upmost monolayer of anti-T antibody via layer-by-layer self-assembly technique. After the fabrication of anti-T antibody multilayers on IOL surface, the surface characteristics of the anti-T antibody functionalized IOL, as well as its effect on LECs adhesion, proliferation, migration and EMT were then tested in this study. Our results revealed that anti-T antibody multilayers could be successfully immobilized onto IOL surfaces by plasma pretreatment and layer-by-layer self-assembly technique, and could keep stable for at least 3 months on IOL surface. The anti-T antibody immobilized in the multilayers on IOL surfaces showed good immunological activity by its specific antigen-antibody interaction with exogenous TGF-β2. Anti-T antibody functionalized IOL surface was as smooth and flat as the untreated IOL surface. No difference in optical or physical properties was found between the anti-T antibody functionalized IOLs and the untreated IOLs. Compared with the untreated IOLs, the anti-T antibody functionalized IOL greatly inhibited LECs from migration and EMT, yet showed only transient inhibition to LECs adhesion and no inhibition to LECs proliferation. With these data, we demonstrate a simple, inexpensive, and feasible method to fabricate surface functionalized IOL for in situ capture and neutralization of TGF-β2 in the capsular bag, which might be a possible solution to preventing posterior capsule opacification after cataract surgery. © 2013 Elsevier B.V.


Jin X.,Zhejiang University | Jin X.,Zhejiang Provincial Key Laboratory of Ophthalmology | Zhao Y.,Zhejiang University | Zhao Y.,Zhejiang Provincial Key Laboratory of Ophthalmology | And 4 more authors.
Ophthalmic Plastic and Reconstructive Surgery | Year: 2014

PURPOSE: To assess the effectiveness of the Crawford tube for the treatment of chronic suppurative lacrimal canaliculitis. METHODS: A prospective study was performed on 8 consecutive patients who accepted the Crawford tube to treat chronic suppurative lacrimal canaliculitis. Postoperatively, the patients were treated with 0.5% levofloxacin eyedrops 4 times per day and oral levofloxacin tablets 0.5 g once per day for 4 days. Follow-up times were >3 months after removing the Crawford tube. The condition of the lacrimal punctum and patient's symptoms were carefully evaluated. RESULTS: All patients had unilateral single canaliculus involvement and had excellent resolution of canaliculitis without the need for surgical treatment. Complications included lacrimal punctum granulation and lacrimal punctum dehiscence. CONCLUSIONS: The insertion of the Crawford tube may offer an alternative to surgery in the management of suppurative lacrimal canaliculitis. © 2014 The American Society of Ophthalmic Plastic and Reconstructive Surgery, Inc.


Mertens F.,Catholic University of Leuven | Gremeaux L.,Catholic University of Leuven | Chen J.,Catholic University of Leuven | Chen J.,Huazhong University of Science and Technology | And 14 more authors.
Endocrine-Related Cancer | Year: 2015

Pituitary adenomas cause significant endocrine and mass-related morbidity. Little is known about the mechanisms that underlie pituitary tumor pathogenesis. In the present study, we searched for a side population (SP) in pituitary tumors representing cells with high efflux capacity and potentially enriched for tumor stem cells (TSCs). Human pituitary adenomas contain a SP irrespective of hormonal phenotype. This adenoma SP, as well as the purified SP (pSP) that is depleted from endothelial and immune cells, is enriched for cells that express 'tumor stemness' markers and signaling pathways, including epithelial-mesenchymal transition (EMT)-linked factors. Pituitary adenomas were found to contain self-renewing sphere-forming cells, considered to be a property of TSCs. These sphere-initiating cells were recovered in the pSP. Because benign pituitary adenomas do not grow in vitro and have failed to expand in immunodeficient mice, the pituitary tumor cell line AtT20 was further used.We identified a SP in this cell line and found it to be more tumorigenic than the non-SP 'main population'. Of the two EMTregulatory pathways tested, the inhibition of chemokine (C-X-C motif) receptor 4 (CXCR4) signaling reduced EMT-associated cell motility in vitro as well as xenograft tumor growth, whereas the activation of TGFb had no effect. The human adenoma pSP also showed upregulated expression of the pituitary stem cell marker SOX2. Pituitaries from dopamine receptor D2 knockout (Drd2K/K) mice that bear prolactinomas contain more pSP, Sox2C, and colony-forming cells than WT glands. In conclusion, we detected a SP in pituitary tumors and identified TSC-associated characteristics. The present study adds new elements to the unraveling of pituitary tumor pathogenesis and may lead to the identification of new therapeutic targets. © 2015 Society for Endocrinology Printed in Great Britain.


Ni S.,Zhejiang University | Ni S.,Zhejiang Provincial Key Laboratory of Ophthalmology | Yu Y.,Zhejiang University | Yu Y.,Zhejiang Provincial Key Laboratory of Ophthalmology | And 8 more authors.
PLoS ONE | Year: 2013

Objectives:The aims of the present study were to determine oxidative stress and to explore possible reasons of reactive oxygen species (ROS) increase in human lens epithelial (HLE) B3 cells exposed to low intensity 1.8 GHz radiofrequency fields (RF).Methods:The HLE B3 cells were divided into RF exposure and RF sham-exposure groups. The RF exposure intensity was at specific absorption rate (SAR) of 2, 3, or 4 W/kg. The ROS levels were measured by a fluorescent probe 2′7′-dichlorofluorescin diacetate (DCFH-DA) assay in the HLE B3 cells exposed to 1.8 GHz RF for 0.5, 1, and 1.5 h. Lipid peroxidation and cellular viability were detected by an MDA test and Cell Counting Kit-8 (CCK-8) assays, respectively, in the HLE B3 cells exposed to 1.8 GHz RF for 6, 12, and 24 h, respectively. The mRNA expression of SOD1, SOD2, CAT, and GPx1 genes and the expression of SOD1, SOD2, CAT, and GPx1 proteins was measured by qRT-PCR and Western blot assays in the HLE B3 cells exposed to 1.8 GHz RF for 1 h.Results:The ROS and MDA levels significantly increased (P<0.05) in the RF exposure group and that the cellular viability, mRNA expression of four genes, and expression of four proteins significantly decreased (P<0.05) compared with the RF sham-exposure group.Conclusions:Oxidative stress is present in HLE B3 cells exposed to 1.8 GHz low-intensity RF and that the increased production of ROS may be related to down-regulation of four antioxidant enzyme genes induced by RF exposure. © 2013 Ni et al.


Lai K.,Zhejiang University | Cui J.,Zhejiang University | Ni S.,Zhejiang University | Zhang Y.,Zhejiang University | And 3 more authors.
PLoS ONE | Year: 2013

Background:Cataract is the leading cause of blindness worldwide. Many observational studies assessed the relationship between postmenopausal hormone replacement therapy (HRT) and risk of cataract development, but the reported results were controversial. The aim of present meta-analysis was to evaluate the association of postmenopausal hormone replacement therapy with risk of cataract development.Methods:The eligible observational studies, including cross-sectional, case-control and cohort studies, were identified by searching PubMed and Embase during March of 2013. Either a fixed- or a random-effects model was used to calculate the pooled odds ratio (OR) with its 95% confidence interval (95%CI). Subgroup analysis on cataract types was performed.Results:A total of four cohort and five case-control or cross-sectional studies were finally included into this meta-analysis. Overall, a significant decreased risk of developing any type of cataract was found in ever HRT group as compared with non-HRT group among cohort studies (OR 0.83; 95%CI: 0.71,0.97) and case-control or cross-sectional studies (OR 0.74; 95%CI: 0.59,0.93). Subgroup analysis on cataract types determined that the significantly decreased risk of nuclear cataract in current HRT group (OR 0.72; 95%CI: 0.61,0.85) and also a critically reduced risk of nuclear cataract in ever HRT group (OR 0.80; 95% CI: 0.64,1.01) were found among case-control or cross-sectional studies, as compared with non-HRT group. No association of HRT with risk of cortical and posterior subcapsular cataract was observed.Conclusions:The results of present meta-analysis indicate that postmenopausal hormone use may play a protective role in cataract development. © 2013 Lai et al.


Wan T.,Zhejiang University | Wan T.,Zhejiang Provincial Key Laboratory of Ophthalmology | Jin X.,Zhejiang University | Jin X.,Zhejiang Provincial Key Laboratory of Ophthalmology | And 5 more authors.
Current Eye Research | Year: 2016

Purpose: To investigate the relationship between meibomian gland dysfunction (MGD) and incomplete blinking caused by cranial nerve seven (CN VII) palsy. Methods: A prospective case series of 60 consecutive patients with unilateral CN VII palsy was evaluated for MGD. According to the House-Brackmann scale, patients were divided into complete or incomplete blinking group, with the incomplete group further subdivided to early, middle or late stage according to the paralysis duration. Schirmer's I test, tear break-up time (BUT), superficial punctate keratopathy, eyelid abnormality, ability and quality of meibum expression were evaluated. Unaffected contralateral eyes were used as control for comparison. Results: A paired sample t-test between affected and unaffected eyes demonstrated a significant difference for BUT, superficial punctate keratopathy, eyelid abnormality, ability and quality of meibum expression in the incomplete blinking group. However, only BUT showed significant difference between affected and unaffected eyes in complete blinking group. Furthermore, we showed that paralysis duration was related to the incidence of MGD. Significant difference was demonstrated between the affected and unaffected eyes for superficial punctate keratopathy, eyelid abnormality, ability and quality of meibum expression in the middle and late stage, but not early stage. Conclusions: These findings suggest that the inability to blink completely induced by CN VII palsy for more than one week might contribute to the development of MGD. Clinical optimization of blinking may ameliorate MGD symptom and benefit CN VII palsy patients. © 2016 Taylor & Francis Group, LLC.


Ye P.,Zhejiang University | Ye P.,Zhejiang Provincial Key Laboratory of Ophthalmology | Liu J.,Zhejiang University | Liu J.,Zhejiang Provincial Key Laboratory of Ophthalmology | And 6 more authors.
International Journal of Medical Sciences | Year: 2014

Objective: miR-126, the miRNA considered to be specially expressed in endothelial cells and hematopoietic progenitor cells, is strongly associated with angiogenesis. The purpose is to evaluate the role of miR-126 in hypoxia-induced angiogenesis and the possible mechanisms. Methods: The expression of miR-126 was detected in hypoxia-treated RF/6A cells and diabetic retinas using real-time PCR. The miR-126 was up- or down-regulated by transfecting miR-126-mimics or inhibitors into RF/6A cells. Cell cycle analysis was performed using flow cytometry. The protein levels of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) were assessed by immunoblotting. Results: A significantly decreased expression of miR-126 was found in hypoxia-treated RF/6A cells in a time-dependent manner compared with normoxic condition. The expression of miR-126 was also reduced in the retina tissue of streptozotocin-induced diabetic rats. The expression of VEGF and MMP-9 proteins was increased in hypoxia-induced RF/6A cells. In the functional analysis, miR-126-mimic significantly reduced the percentage of RF/6A cells in S phases compared with the negative control under hypoxic conditions. Furthermore, the VEGF and MMP-9 protein levels were sharply decreased in hypoxia-induced RF/6A cells pretreated with miR-126-mimics and increased in the cells pretreated with miR-126-inhibitors. Conclusions: miR-126 is down-regulated under hypoxic condition both in vitro and in vivo and may halt the hypoxia-induce neovascularization by suspending the cell cycle progression and inhibiting the expression of VEGF and MMP-9. © Ivyspring International Publisher.


Yu Y.,Zhejiang University | Yu Y.,Zhejiang Provincial Key Laboratory of Ophthalmology | Chen P.,Zhejiang University | Li J.,Zhejiang University | And 7 more authors.
BMC Medical Genetics | Year: 2014

Background: The major intrinsic protein gene (MIP), also known as MIP26 or AQP0, is a member of the water-transporting aquaporin family, which plays a critical role in the maintenance of lifelong lens transparency. To date, several mutations in MIP (OMIM 154050) have been linked to hereditary cataracts in humans. However, more pathogenic mutations remain to be identified. In this study, we describe a four-generation Chinese family with a nonsense mutation in MIP associated with an autosomal dominant congenital cataract (ADCC), thus expanding the mutational spectrum of this gene. Methods: A large four-generation Chinese family affected with typical Y-suture cataracts combined with punctuate cortical opacities and 100 ethnically matched controls were recruited. Genomic DNA was extracted from peripheral blood leukocytes to analyze congenital cataract-related candidate genes. Effects of the sequence change on the structure and function of proteins were predicted by bioinformatics analysis. Results: Direct sequencing of MIP in all affected members revealed a heterozygous nucleotide exchange c.337C>T predicting an arginine to a stop codon exchange (p.R113X). The substitution co-segregated well in all the affected individuals in the family and was not found in unaffected members or in the 100 unrelated healthy controls. Bioinformatics analysis predicted that the mutation affects the secondary structure and function of the MIP protein. Conclusions: We identified a novel mutation of MIP (p.R113X) in a Chinese cataract family. This is the first nonsense mutation of MIP identified thus far. This novel mutation is also the first disease-causing mutation located in the loop C domain of MIP. The results add to the list of mutations of the MIP linked to cataracts. © 2014 Yu et al.; licensee BioMed Central Ltd.

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