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Research progress of the related cytokines of Th1 and Th2 on glaucoma

Chinese Journal of Experimental Ophthalmology.2017;35(11):1052-1056. doi:10.3760/cma.j.issn.2095-0160.2017.11.020

Glaucoma is a leading cause of irreversible blindness,which includes visual field defects and chronic degenerative diseases of the optic nerve,such as apoptosis of the retinal ganglion cells (RGCs) and progressive loss of optic nerve axons.Elevated intraocular pressure is the primary risk factor for glaucoma,and lowering intraocular pressure is the first choice of treatment for slowing the development of glaucoma.It has recently been found that immunological factors are one of the non-stress-dependent risk factors for optic nerve damage in glaucoma,while most of the immune and non-immune biological effects are regulated by cytokines.CD4+ helper T cells are the major sources of cytokines.Thl and Th2-related cytokines play an essential role in the pathogenesis of glaucoma and related with the survival and apoptosis of RGCs.In this review,we discussed the potential relationship between Th1/Th2-related cytokines and glaucoma.

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Is microbial infection always harmful?

Wenna GAO

Chinese Journal of Experimental Ophthalmology.2017;35(11):1047-1051. doi:10.3760/cma.j.issn.2095-0160.2017.11.019

It is possible for microbial infection (including parasitic infection) to disrupt the balance of autoimmune tolerance,result in the dysfunction of immune system,and make the organism encountered with the attack of the autoimmune disease.As one of the common places between foreign antigens and autoantigens,molecular mimicry is believed to play a vital role in the pathogenesis of autoimmune diseases.However,a large number of studies suggest that rather than inducing various symptoms of autoimmune diseases,many infection of microorganisms covered with mimic peptide of the autoantigen may even protect the infected organism from the disease at the level of antibodies or T cells when the analogous antigen invaded again.The present paper reviews the possible prognosis after microbial infection based on molecular mimicry.

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Research progress of internal limiting membrane peeling technique

Dong FANG

Chinese Journal of Experimental Ophthalmology.2017;35(11):1039-1042. doi:10.3760/cma.j.issn.2095-0160.2017.11.017

Internal limiting membrane (ILM) peeling technique has been widely used in the treatment of a variety of vitreoretinal diseases for the advantage of releasing vitreomacular traction.However,this routine approach was less effective in refractory macular disorders.Meanwhile,the technique and the use of stains could lead to structural and functional changes of macular.Therefore,to better preserve macular structure,modified ILM peeling techniques including inverted ILM flap technique,autologous transplantation of ILM technique and the foveola nonpeeling ILM technique emerged.Recent researches had demonstrated favorable structural and functional outcomes of these techniques on giant macular hole,refractory macular hole and high myopic macular hole.However,more longterm follow-up studies with larger samples are needed to confirm the current results.In this paper,we reviewed the progress of modified ILM peeling technique in vitreoretinal diseases.

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Influence of air pollution on ocular surface

Qichen YANG

Chinese Journal of Experimental Ophthalmology.2017;35(11):1035-1038. doi:10.3760/cma.j.issn.2095-0160.2017.11.016

Air pollution is the over emission of harmful materials into atmosphere,causing damage to human beings and other organisms.Some researches indicate that air pollution can damage ocular surface,change the tear's ingredients,and cause some symptoms,such as red eyes,lacrimation,foreign body sensation and so on.It can also change the components of the tear and damage the epithelial cells of the eyes.The mechanisms including the direct effects of the pollutant,the secondary changes of the tear,the inflammation and the oxidative stress response.As the growing problems of air pollution,its influence on ocular surface has been attracting more and more attentions.How to prevent the damage caused by air pollution is the question that ophthalmologists should think about in the future.In this paper,we focus on the sources and ingredients of air pollution,the changes and the pathogenesis of ocular surface caused by air pollution and the summary of the studies of air pollution on the ocular surface in the past few years.Studying the influence of air pollution on ocular surface significantly contributes to the theoretical research and clinical prevention.

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Association analysis of advanced glycation end-products in lens with diabetic retinopathy severity

Zhuoshi WANG ; Qianru ZHANG ; Pingli ZHU ; Liangbao AN ; Wei HE

Chinese Journal of Experimental Ophthalmology.2017;35(11):1030-1034. doi:10.3760/cma.j.issn.2095-0160.2017.11.015

Background The deposition of advanced glycation end products (AGEs) in lens is the risk factor of diabetic complications.Researches revealed that AGEs has autofluorescence.Crystallin is a longevity protein.AGEs accumulation is probably associated with diabetic retinopathy (DR).Objective This study was to evaluate the association of AGEs autofluorescence intensity with diabetes and with DR.Methods A cross-sectional study was carried out under the approval of Ethic Committee of Shenyang He Eye Hospital and informed consent of each patient.One hundred eyes of 100 patients with age-related cataract aged 50-70 years were included in He Eye Hospital from September to December 2015.The patients were divided into non-diabetes group (40 patients) and diabetes group (60 patients),and then the patients in diabetes group were subdivided into non-DR (NDR) group,non-proliferating DR (NPDR) group and proliferating DR (PDR) group according to the DR grading criteria,20 patients for each.Glycosylated henoglobin A1c (HbA1c) and fasting plasma glucose (FPG) were detected for each subject,and the lens autofluorescence was assayed with lens fluorescence biomicroscope (Clearpath DS120).The association of lens autofluorescence intensity with serum HbA1c level or DR severity was analyzed.Results The age and diabetes course were matched among the non-diabetes group,NDR group,NPDR group and PDR group (F=2.587,2.899,both at P＞0.05),and the FBS and HbA1c level were evidently higher in the NDR group,NPDR group and PDR group than those in the non-diabetes group (all at P＜0.01).The autofluorescence intensity of lens was (0.159±0.032),(0.256±0.024),(0.319 ±0.013) and (0.394±0.035) cd in the non-diabetes group,NDR group,NPDR group and PDR group,respectively,showing a significant difference among the groups (F =90.265,P =0.000).The autofluorescence intensity of lens in the NDR group,NPDR group and PDR group was significantly increased in comparison with the non-diabetes group and the autofluorescence intensity of lens was gradually increased with the severity of DR (all at P＜0.01).A positive linear correlation was found between autofluorescence intensity of lens and serum HbA1 c level in diabetes patients (r =0.654,P ＜ 0.05).Conclusions The autofluorescence intensity of AGEs in lens appears to be associated with the severity of DR and HbA1 c.The autofluorescence intensity of AGEs in the lens of diabetes patient is probably one of the evaluation indexes of early stage of DR.

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Adjuvant treatment of neovascular glaucoma with conbercept: clinical efficacy and histopathological analysis of trabeculectomy specimens

Liukun SHI ; Jin YANG ; Jinyong LIN

Chinese Journal of Experimental Ophthalmology.2017;35(11):1019-1024. doi:10.3760/cma.j.issn.2095-0160.2017.11.013

Background Neovascular glaucoma (NVG) often occurs secondary to ocular ischemic diseases.Intravitreal injection of bevacizumab or ranibizumab as an adjunct therapy or NVG is safe and efficient.However,the efficacy of intravitreal injection of conbercept for NVG is still under exploration.Objective This study was to evaluate the efficacy and safety of intravitreal conbercept combined with trabeculectomy and panretinal photocoagulation for NVG.Methods A prospective,interventional case series study was performed.Twenty-six patients (27 eyes) who were diagnosed with NVG in Tianjin Eye Hospital were included.Twenty-five eyes were initially injected with conbercept into vitreous,then trabeculectomy and panretinal photocoagulation were performed,the best corrected visual acuity (LogMAR),intraocular pressure and treatment-related indexes were examined before and after treatment,the intraoperative and postoperative complications were evaluated.Light microscopy and transmission electron microscopy were used to observe the surgically excised trabecular tissue.Enucleation was performed on 2 eyes,and the surgically excised tissue was taken as the histopathological controls.Results At six monthes after surgery,the intraocular pressure of the 25 patients with intravitreal injection of conbercept were decreased from (48.8 ± 10.2) mmHg to (18.0±4.3) mmHg,the number of anti-glaucoma medications were decreased from 3.0 (3.0,4.0) to 0.0 (0.0,1.0) (both at P＜0.05).Best corrected visual acuity (LogMAR) was increased from 2.0± 0.9 to 1.5 ± 1.4 (P＞0.05).Twenty-one eyes displayed controlled intraocular pressure (≤ 21 mmHg) after treatment.Regression of neovasculariztion in the surface of iris and thin-walled vessels in the iris stroma in specimens with intravitreal injection of conbercept were revealed by light microscopy.New vessels decreased,endothelial cell degeneration and vascular occlusion or cavity in the iris stroma in the conbercept-treatment specimens were revealed by transmission electron microscopy.Conclusions A comprehensive therapy for NVG can regress iris neovascularization,reduce postoperative complication,effectively control intraocular pressure and preserve visual function by intravitreal injection of conbercept,anti-glaucoma surgery and panretinal photocoagulation.

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Clinical efficacy evaluation of vitrectomy combined with autologous internal limiting membrane transplantation for large macular hole

Yuhua HAO ; Ruijie XI ; You HAN ; Xiaoyu TIAN ; Li DAI ; Junfang SHI

Chinese Journal of Experimental Ophthalmology.2017;35(11):1014-1018. doi:10.3760/cma.j.issn.2095-0160.2017.11.012

Background Giant idiopathic macular hole (IMH) severely affects visual acuity and increases operative difficulty during the surgery,so modifying or optimizing the operation method is helpful for improving the prognosis.Objective This study aimed to evaluate the efficacy and safty of vitrectomy combined with free internal limiting membrane transplantation for large IMH.Methods A prospective serial cases-observational study was carried out under the informed consent of each patient.Forty-two eyes of consecutive 42 patients with IMH of mean diameter (814.31±112.95)μm were included in the Second Hospital of Hebei Medical University from January 2013 to November 2015.All the eyes received vitrectomy combined with free internal limiting membrane transplantation and 12％ C3F8 filling.The best corrected visual acuity (BCVA) (LogMAR),inner segment/outer segment (IS/OS) defect range,external limiting membrane defect range,retinal thickness at macular fovea were measured with slit lamp microscope,indirect ophthalmoscope and spectral-domain optical coherence tomography (SD-OCT) before surgery and 1,3,6 and 12 months after surgery.Results IMH complete closure in 97.6％ eyes (41/44) at 12 months after surgery.BCVA was improved after operation and showed a significant difference among various time points (F =28.032,P＜0.001).The IS/OS defect range was (1 112.00±45.44),(859.00±84.55),(649.00±52.47),(486.00±46.88) and (320.00±45.13) μm before surgery and 1,3,6 and 12 months after surgery,showing a significant difference among different time points (F=38.761,P＜0.001),and the IS/OS defect range was gradually shrinked after operation compared with that before operation (all at P＜0.05).The mean defect range of external limiting membrane was (1 038.00 ±39.63),(748.00±64.12),(585.00±48.88),(438.00±42.84) and (265.00±28.97)μm before surgery and 1,3,6 and 12 months after surgery,with a significant difference among various time points (F=36.459,P＜0.001),and the mean defect range of external limiting membrane was evidently reduced (all at P＜0.05).The foveal retinal thickness value was increased at 3,6 and 12 months after surgery,which was significantly higher than that 1 month after surgery (all at P＜0.05).The high reflect signal was faded away at 3 months after operation,indicating that implanted internal limiting membrane was decomposed and metabolized.Conclusions Vitrectomy combined with autologous internal limiting membrane transplantation seems to be safe and effective for large IMH.

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Promoting effects of macrogol 15 hydroxystearate micelles on corneal absorption of terbinafine hydrochloride

Tianyang ZHOU ; Huiyun XIA ; Jijun HE ; Junjie ZHANG

Chinese Journal of Experimental Ophthalmology.2017;35(11):1009-1013. doi:10.3760/cma.j.issn.2095-0160.2017.11.011

Background Macrogol 15 hydroxystearate (HS15) is a novel soft non-ionic surfactant and is widely used to solubilize the poorly soluble drugs due to high drug loading and enhancing permeability ability to hydrophobic drug.However,the delivering effects to cornea of HS15 on terbinafine hydrochloride (TH),a insoluble antifungal agents,is unclear.Objective This study was to investigate the promoting corneal absorption effects of HS15 micelles (HNMs) on TH.Methods TH-HNMs was prepared by a co-solvent method.The hydrodynamic droplet size,polydispersity index,and Zeta potential of TH-HNMs were measured by using a Zetasizer.The shape of the micelles was observed under the transmission electron microscope.The high performance liquid chromatography (HPLC) was employed to detect the in vitro cumulative releasing level of TH in the TH-HNMs and drug entrapment efficiency.TH-HNMs was topically adninistered in eyes of 5 healthy male New Zealand rabbits to evaluate the ocular irritation response.Ninety rabbits were randomized into experimental group and control group,and 50 μl of 0.5％ TH-HNMs and 0.5％ oily TH were topically administered in the right eyes of the animals in the experimental group and contral group,respectively.The animals were sacrificed 5,15,30,60,90,120,180,240,360 minutes after eye dropping by over anesthetization way and the corneas were harvested,and the TH content in the cornea was detected using HPLC.The study protocal was approved by Life Science Ethic Committee of Henen Eye Hospital.Results The average size and polyolis persital index of TH-HNMs were 13.32 nm and 0.046,respectively,and its average Zeta potential was-0.133 mV.The drug entrapment efficiency was 100％.The release level of TH from the micelles presented a pH-dependent manner.The release level of TH was (95.20±3.20)％ in the phosphate buffer with pH 5.0 and (0.17± 0.01)％ in the phosphate buffer with pH 7.4.The ocular irritation score was 2,and no visible damage was found around experimental eyes after instillation of TH-HNMs.The peak content of TH in the rabbit cornea 5 minutes was (20.26±2.26)pg/g in the experimental group,which was significantly higher than (1.40± 0.44)μg/g in the control group (t =18.926,P=0.000).The area under the curve (AUC)0.360min of drug concentration-time curve in the experimental group was 1 292.25 μg/(g · min),which was 15.6 times more than the control group.Conclusions TH-HNMs is an ideal agent with a simple preparing process,high drug entrapment efficiency,small size and low ocular irritation.Compared with oily TH,TH-HNMs can effectively enhance the corneal absorption of TH.

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Study on the relationship between CFTR physiological secretion pathway and intracellular calcium signaling in rabbit cornea epithelium

Limian LIN ; Yiling LUO ; Shiyou ZHOU

Chinese Journal of Experimental Ophthalmology.2017;35(11):998-1002. doi:10.3760/cma.j.issn.2095-0160.2017.11.009

Background Researches showed that cystic fibrosis transmembrane conductance regulator protein(CFTR) is a channel secreting anion and water,and it plays an important role in tear secreting.Traditional conception thought that CFTR pathway is cAMP-PKA-dependent without the participation of intracellular calcium.However,studies disclosed that elevating intracellular cAMP could not open the CFTR channel.So whether calcium signal is associated with CFTR-related corneal epithelial secretion is controversial.Objective This study was to investigate the association between CFTR secretion and intracellular calcium signaling in rabbit corneal epithelium.Methods Sixteen New Zealand white rabbits were randomized into odd number group and even number group by computer randomized number method.The corneas were obtained under the general anesthesia and placed in Ussing Chamber for the record of short cricuit current (Isc).The right eyes of rabbits in the odd number group were stimulated with ATP and served as ATP stimulating group.The left eyes were pretreated with CFTRinh-172 prior to ATP stimulation and served as CFTRinh-172 pretreated group.In the even number group,the left eyes of rabbits were pretreated with BMPTA/AM before ATP stimulation and served as BMPTA/AM pretreated group,and the right eyes of the rabbits were used to isolate and culture corneal epithelial cells by explant adherent method,the level of intracellular calcium were evaluated using Leica SP5 laser scan confocal microscope.Results The ATP-induced ΔIsc of corneal epithelium was (5.73 ± 1.36),(1.30 ± 0.95) and (2.47 ± 0.55) μA/cm2 in the ATP stimulating group,CFTRinh.172 pretreated group and BMPTA/AM pretreated group,respectively,and the AIsc was significantly reduced in the CFTRinh.172 pretreated group and BMPTA/AM pretreated group compared with ATP stimulating group (t=11.201,5.508,both at P ＜ 0.001).The fluorescence intensity of intracellular calcium release after ATP stimulation was 3.25 folds more than that before ATP stimulation.Conclusions ATP promotes rapid short circuit current of corneal epithelium.CFTRinh-172 depresses the ATP-induced corneal epithelium AIsc,and BMPTA/AM suppresses intracellular calcium release.It is suggested that intracellular calcium signaling secretion probably participates in the functional CFTR activity in rabbit corneal epithelium.

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The inhibitory effects of baicalein on overexpression of proline hydroxylase 2 in hypoxia-induced retinal glial cells

Ning YANG ; Qiang LU ; Mengke YUAN ; Han ZHANG ; Wei CUI

Chinese Journal of Experimental Ophthalmology.2017;35(11):990-996. doi:10.3760/cma.j.issn.2095-0160.2017.11.007

Background Proline hydroxylase 2 (PHD2) is a degrading enzyme of hypoxia-inducible factor-1α(HIF-1α) and can regulate the expression of vascular endothelial growth factor (VEGF) and erythropoietin that promote hypoxia-induced blood vessel generation.Baicalein is an inhibitor of PHD2.Understanding the inhibitory effects of baicalein on overexpression of PHD2 in hypoxia-induced retinal glial cells is helpful for elucidating the pathogenesis of the ocular nevascular diseases.Objective This study aimed to observe the expression change of PHD2 in hypoxia-induced retinal glial cells and explore the inhibition of baicalein on PHD2.Methods The study protocol was approved by Experimental Animal Ethic Committee.Retinal glial cells were isolated from SPF neonatal SD rats (within 48 hours) and primarily cultured in DEME by explant cell culture.Cultured cells were identified by immunofluorescence staining of glial fibrillary acidic protein.The cells were divided into normal control group,CoCl2 group and CoCl2 +baicalein group.The cells in the normal control group were cultured in the conventional medium,and 200 μmol/L CoCl2 was added in the medium to establish the hypoxia cell models in the CoCl2 group,and 200 μmol/L CoCl2 and 50 μmol/L baicalein were added in the medium in the CoCl2 + baicalein group.The proliferation (absorbancy) of the cells was detected by cell counting kit-8 (CCK-8).The expression of PHD2 and VEGF in the cells was detected and located using immunofluorescence staining.The expressions of PHD2 mRNA and VEGF mRNA and their proteins in the cells were assayed by real-time quantitative PCR and Western blot,respectively.Resnlts The proliferation (absorbancy) of the cells was significantly different among the groups (F=132.05,P=0.00),and the absorbancy was considerably increased in the CoCl2 group compared with normal control group and CoCl2 + baicalein group,with significant differences between the groups (both at P＜ 0.01).Immunofluorescence staining showed that the PHD2 and VEGF were weakly expressed in the cells of the normal control group,and the expression intensity was high in the CoCl2 group.The expression intensities of PHD2 and VEGF in the cells were weakened in the CoCl2 +baicalein group compared with those of the CoCl2 group.The relative expression levels of PHD2 mRNA were 0.366±0.034,0.894 ± 0.015 and 0.445 ± 0.017 and those of PHD2 protein were 131.27 ± 2.61,140.12 ± 4.29,133.14±2.11;those of VEGF mRNA were 1.346 ± 0.008,3.465 ± 0.048 and 2.264 ± 0.073 and those of VEGF protein were 532.25 ± 19.92,601.13 ± 10.21,537.34 ± 5.96 in the normal control group,CoCl2 group and CoCl2 + baicalein group,showing significant differences among the three groups (PHD2:F =905.89,43.18,both at P＜0.01.VEGF:F=27.13,185.79,both at P ＜ 0.01),and the relative expression levels of PHD2 and VEGF mRNA and protein in the CoCl2 group were higher than those in the normal control group and CoCl2 +baicalein group (all at P＜ 0.01).Conclusions Hypoxia stimulates the proliferation of retinal glial cells and increase the expression of PHD2 and VEGF.Baicalein can effectively inhibit the overexpression of PHD2 and VEGF in retinal glial cells and proliferation of retinal glial cells induced by hypoxia.PHD2 is a potential provention and treatment target of hypoxiainduced retinal neovascularization.

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