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Objective:To explore the changes in morphology and function of meibomian gland and the expressions of inflammatory factors and lipid metabolic factors in meibomian gland of diabetic mice.Methods:Fifty 8-week-old male C57BL/6 mice of clean degree were divided into normal control group ( n=20) and diabetes model group ( n=30) according to a random table.Diabetes model was established by the intraperitoneal injection of streptozotocin (60 mg/kg, 10 mg/ml). Mouse tail vein blood glucose ≥16.7 mmol/L was considered as successful modeling.Blood glucose was measured weekly, and body weight was compared between the two groups.Ten mice were randomly selected for fluorescein sodium staining of the cornea to evaluate the integrity of the corneal epithelium from both groups at an interval of 4 weeks.Five mice were randomly selected from the two groups and were sacrificed via anesthesia to collect meibomian gland tissue for hematoxylin and eosin staining in order to observe morphological changes at 8 and 16 weeks after modeling, respectively.At 16 weeks following modeling, mebomian gland of 5 mice randomly selected from both groups was stained with oil red O staining to observe the distribution of lipid.Real-time fluorescence quantitative-PCR was performed to detect the relative expressions of tumor necrosis factor (TNF)-α, pigment epithelium derived factor (PEDF), peroxisome proliferators-activated receptor γ (PPARγ), and adipose differentiation-related protein (ADFP) mRNA in meibomian gland.The use and care of animals complied with the ARVO statement.This study protocol was approved by the Institutional Animal Care and Use Committee of Tianjin Medical University Eye Hospital (No.TJYY20190630009). Results:The successful modeling rate of diabetes in mice was 100%, and the survival rate was 83.3% (25/30). The weight was significantly lower and the blood glucose level was higher in diabetes model group at 8 and 16 weeks after modeling in comparison with normal control group (all at P<0.05). There were significant differences in corneal fluorescein staining score among different time points in diabetes model group ( F=27.155, P<0.05). In diabetes model group, thinner wall of meibomian gland duct, enlarged lumen of the duct, dilated acini and oil red-stained lipid deposition in most acini were observed.At 16 weeks after modeling, the expressions of TNF-α, and PPARγ mRNA in meibomian gland of diabetes model group were 3.33±0.91 and 1.55±0.25, which were significantly higher than 1.00±0.16 and 1.00±0.27 of normal control group (both at P<0.05). The expression of PEDF mRNA in diabetes model group was 0.42±0.08, which was significantly lower than 1.00±0.34 in normal control group ( P<0.05). There was no significant difference in the ADFP mRNA expression between the two groups ( t=0.943, P=0.38). Conclusions:Inflammatory factors and lipid metabolic factors such as TNF-α, PEDF, and PPARγ may be involved in the pathogenesis of meibomian gland dysfunction induced by diabetes.
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Exosomes are membranous vesicles secreted by cells and can be widely involved in intercellular communication, anti-inflammation, immunoregulation, etc.Mesenchymal stem cell (MSC), regulatory T cell (Treg), immature dendritic cell (imDC) and myeloid-derived suppressor cell (MDSC) are the main ocular surface-related exosomes origins.Exosomes derived from different cells play their roles by delivering different biological molecules to recipient cells.Exosomes derived from MSC play a positive role in ocular surface inflammation and immune-related diseases by inhibiting T cell proliferation, transforming macrophage phenotype, regulating T helper (Th) cell differentiation and up-regulating Treg expression, reduce neovascularization and inflammation, and foster a microenvironment to promote corneal wound healing at the same time.Exosomes derived from Treg contain inducible NO synthase and microRNA (miRNA) including miR-503, miR-330 and miR-9, which can interfere with cell cycle progression, induce apoptosis, induce the differentiation of other T cells into Treg phenotype, inhibit T cell allograft rejection to induce immune tolerance.Exosomes derived from imDC inhibit corneal allograft rejection by delivering miR-682.MDSC-derived exosomes promote Treg expansion in vivo and in vitro, inhibit the proliferation and cytotoxicity of activated T cells, and express miR-29a-3p and miR-93-5p, which can inhibit the differentiation of Th1 and Th17 cells.Given the anti-inflammatory and immunosuppressive effects of exosomes, this paper reviewed the studies on ocular surface inflammation and immune-related diseases such as corneal injury, mucopolysaccharide storage disease, dry eye, Sj?gren syndrome and ocular graft-versus-host disease.
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Objective:To explore the damage of panretinal photocoagulation (PRP) to the subbasal nerve plexus (SNP) and its related mechanisms by comparing SNP changes in wide-field mosaic between before and after PRP treatment in diabetic patients.Methods:A randomized controlled study was conducted.Fifty-seven patients (114 eyes) with type 2 diabetes mellitus and binocular diabetic retinopathy (DR) stage IV to receive PRP treatment in Shanxi Eye Hospital from April to November 2019 were enrolled.The subjects were randomly divided into horizontal-vertical laser group and vertical-horizontal laser group according to a random number table.Twenty-nine eyes from 29 patients were assigned to the horizontal-vertical laser group with the photocoagulation sequence of temporal-nasal-inferior-superior.Twenty-eight eyes from 28 patients were assigned to the vertical-horizontal laser group with the photocoagulation sequence of inferior-superior-temporal-nasal.The severer eyes of each subject were chosen as the treatment eye and the contralateral eyes were chosen as the control eye.Corneal confocal laser scanning microscopy (CCM) was performed before PRP treatment, 1 week after each photocoagulation, and 1 month after the completion of PRP treatment to collect images of the SNP over an area of 2-3 mm around the whorl-like pattern.Captured images at each time were merged into one image by using the Photoshop CC 2017 image processing software, and then the nerve fiber length (NFL) of whorl-like pattern was measured by Neuron J image analysis software.McGill pain questionnaire was used to investigate the pain of patients after each photocoagulation.The NFL changes of SNP at different time points were compared between different eyes and different photocoagulation sequence groups.The study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Shanxi Eye Hospital (No.201804b). Written informed consent was obtained from each patient prior to entering the study cohort.Results:After PRP treatment, there were different degrees of neural structure loss of SNP nerve fibers in 11 treatment eyes, but there was no significant change in SNP nerve fibers in the control eyes.There were significant differences in NFL between the treatment eyes and the control eyes at various time points ( Feyes=2.020, P=0.039; Ftime=4.062, P=0.001). In the horizontal-vertical laser group, different degrees of neural structure loss on the photocoagulation side were found in SNP nerve fibers after the first and second photocoagulation.In the vertical-horizontal laser group, different degrees of neural structure loss on the photocoagulation side were found in SNP nerve fibers after the third and fourth photocoagulation.There was no significant difference in NFL of treatment eyes between the two groups ( Fgroup=0.099, P=0.754), but there was a significant difference in NFL at various time points before and after treatment ( Ftime=5.231, P<0.001). There were 9 (9/57) patients who complained of pain after PRP, which occurred at the first time of photocoagulation in 7 of them. Conclusions:SNP damage may occur after PRP in patients with DR, and SNP is prone to be damaged on the photocoagulation side when performing horizontal photocoagulation.
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Objective:To compare the changes of corneal asphericity and higher-order aberrations after smart pulse technology-assisted transepithelial photorefractive keratectomy (Smart) for low and moderate myopia and to investigate the changes in the shape of the front corneal surface in patients with different diopters.Methods:A non-randomized controlled study design was used.Ninety-eight eyes of 54 patients with moderate or low myopia who underwent Smart surgery in Tianjin Medical University Eye Hospital from November 2018 to March 2019 were included.The 41 eyes of 23 patients with low myopia were set as the low-myopia group, and 57 eyes of 31 patients with moderate myopia were assigned as the moderate-myopia group.The Pentacam anterior segment analysis system was used to measure Q value, index of surface variance (ISV), corneal higher-order aberration (HOA), corneal vertical coma (Z 3-1), corneal horizontal coma (Z 31) and spherical aberration (Z 40) before surgery, 1 month and 3 months after surgery.The anterior surface morphology was compared between the low-myopia and moderate-myopia group.Pearson correlation analysis was used to analyze the correlations between measurement parameters.The study protocol was approved by an Ethics Committee of Tianjin Medical University Eye Hospital (No.2019KY-17). Written informed consent was obtained from each patient before surgery. Results:Corneal Q value, ISV, HOA and Z 40 were 0.445±0.191, 26.973±5.611, 0.671±0.142 and 0.384±0.188, respectively, in the low-myopia group at one month after surgery, which were significantly increased than corresponding preoperative values of -0.273±0.817, 13.784±2.376, 0.433±0.687 and 0.231±0.062 (all at P<0.05). Corneal Q value, ISV, HOA and Z 40 were 0.693±0.203, 34.038±5.773, 0.874±0.216 and 0.520±0.129, respectively, in the moderate-myopia group at one month after surgery, which were significantly increased than corresponding preoperative values of -0.309±0.104, 14.838±3.992, 0.409±0.081 and 0.228±0.089 (all at P<0.05). Corneal Q values, ISV, HOA and Z 40 in the moderate-myopia group were higher than those in the low-myopia group at different time points after surgery, showing significant differences between the two groups (all at P<0.05). There was no significant difference in postoperative 1-month and 3-month corneal Z 3-1 and Z 31 between the two groups (both at P>0.05). The results of correlation analysis showed that there were no significant differences in ΔQ value and ΔISV between the two groups, both of which were negatively correlated with spherical equivalent (ΔQ value: low-myopia group: r=-0.364, P=0.044; moderate-myopia group: r=-0.589, P<0.01; ΔISV: low-myopia group: r=-0.298, P=0.039; moderate-myopia group: r=-0.409, P=0.022). ΔQ value and ΔZ 40 were positively correlated in the moderate-myopia group ( r=0.348, P=0.009); there was no significant correlation between ΔQ value and ΔZ 40 in the low-myopia group ( r=0.180, P=0.266). Conclusions:The corneal high-order aberrations and ISV after Smart are increased in comparison with preoperative values in the low-myopia and moderate-myopia eyes, and the corneal Q values change from negative to positive.The effect of Smart on corneal asphericity is less in the low-myopia eyes.
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Objective:To estimate the correlation between DR and diabetic corneal nerve damage by comparing changes of corneal sub-basal nerve plexus (SNP) in the whorl-like region in different DR stages.Methods:In this cross-sectional study, 66 eyes of 66 type 2 diabetes patients aged 50-65 years old in Shanxi Eye Hospital from September 2018 to January 2019 were included.The subjects were divided into non-diabetic retinopathy group (NDR), non-proliferative diabetic retinopathy group (NPDR) and proliferative diabetic retinopathy group (PDR) accordingly, the right eyes were enrolled.20 eyes of 20 age and gender matched age-related cataract patients were also included as control group.All the subjects underwent confocal corneal microscopy to observe the morphology of SNP in the whorl-like region and measure the nerve fiber length (NFL) in this area.Covariance analysis using disease course as the covariate was used to analyze the differences of NFL value among groups, and Spearman rank correlation analysis was used to analyze the correlation between NFL value and DR stages.The study protocol was approved by the Ethics Committee of Shanxi Eye Hospital.Written informed consent was obtained from each patient prior to entering the study cohort.Results:The whorl-like region were observed in all the eyes.In the control group, nerve fibers were uniform in thickness, densely distributed with regular course, and most of the tested eyes had intact whorl-like structure.Nerve fibers in different DR groups all showed generally decrease in diameter and density, with increase in tortuosity, accompanied by different degrees of whorl-like structure loss.Eyes in the NDR group and NPDR group mainly showed nerve structure loss in the whorl-like center; while some eyes in the PDR group also showed nerve structure loss in the inferior and temporal of the whorl-like region.The NFL values were (21.08±4.74), (16.47±6.35), (14.95±3.90), and (11.61±3.24)mm/mm 2 in the control group, the NDR group, the NPDR group, and the PDR group, respectively, the overall difference was statistically significant ( F=10.541, P<0.001). The NFL value in the control group was significantly higher than those in the different DR groups, the differences were statistically significant (all at P<0.01). Spearman rank correlation analysis showed that there was a significant negative correlation between NFL value and DR stages ( rs=-0.356, P=0.003). Conclusions:Diabetic SNP damage occurs earlier than DR, and progress gradually with the development of DR; Different stages of DR are accompanied by different degrees of nerve structure loss; The structure loss begins at the whorl-like center and gradually develops to downward and the temporal side.The ophthalmologists should take the condition of ocular surface into account when treating ocular fundus diseases of diabetic patients.
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Objective:To evaluate the clinical efficacy and safety of modified chitosan eye drops on rabbit Candida albicans keratitis model.Methods:Ten healthy female New Zealand rabbits were used to establish the superficial Candida albicans keratitis model by the corneal surface lens method in the right eye.Slit lamp microscopy and corneal scraping and microscopic examination were performed to preliminarily determine whether the keratitis model has been successfully established, the rabbits were then randomly divided into a model group and a modified chitosan group by the random number table method.The successfully established rabbit models which were determined by fungal culture results were retained.Five normal rabbits receiving no intervention served as a normal control group.The experimental eyes in the normal control and modified chitosan groups were treated with modified chitosan eye drops, Six times a day for one week, and subsequently four times a day for one week.No treatment was administered to the model group.The changes of corneal lesions and ocular surfaces were examined by slit lamp microscopy every day.At 1, 7, 14, 21 and 28 days after modeling, the eye condition and corneal clinical scores was assessed by slit lamp microscopy.The corneal conditions in each group was observed for two weeks after drug withdrawal.Results:The corneal scraping and microscopic examination results of eight rabbits models showed that the fungal hyphae and spores were positive.The fungal culture results showed that the separated pathogen was Candida albicans.The success rate of modeling was 80%(8/10). The clinical scores in the model group at 7, 14 and 21 days after modeling were 14.50±0.58, 6.25±0.50 and 2.50±0.58, respectively, and were significantly higher than 7.25±1.26, 2.75±0.50 and 1.25±0.50 in the modified chitosan group (all at P<0.05). In the model group, corneal edema was significantly aggravated, and the central white ulcer area was enlarged within seven days after modeling.Between 7 and 28 days after modeling, the corneal ulcer was gradually healed, while the central corneal scar and neovascularization were remained.The average healing time was (24.5±2.6)days.In the modified chitosan group, the corneal infiltration was significantly alleviated within seven days after modeling, and the fungal hyphae and spores of corneal scraping were negative on the 14th day after modeling.The average healing time in the modified chitosan group was (13.5±1.3)days, which was significantly shorter than that in the the model group (P<0.01). No recurrence of keratitis was observed in the modified chitosan group after two weeks of drug withdrawal.The cure rate was 100%.In the normal control group, the conjunctival hyperemia, corneal edema, and lesions were not observed during topical administration.Conclusion:The treatment with modified chitosan eye drop is effective in a rabbit superficial Candida albicans keratitis model, and have no obvious toxic effects on ocular tissues.
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An expert consensus about the clinical diagnosis and treatment of dry eye was documented in 2013 by a corneal expert group of Chinese Ophthalmological Society.However, due to the rapid development of diagnostic and therapeutic devices of dry eye, researoh on dry eye has made significont progress in China since then.Consequently, the existing expert consensus cannot meet the needs of clinical practice.It is therefore urgent to develop a series of standardized diagnosis and treatment protocols, and publish a new consensus of experts and an operating guideline.At the same time, basic, clinical, and translational research on dry eye should be promoted to provide better services to the patients with dry eyes.On January 12, 2019 many experts in the field of dry eye in China held a panel discussion of dry eye study in Guangzhou to analyze the current development status and trends in the field of dry eye in China and abroad.In that meeting, opinions and recommendations were put forward based on a new understanding of the definition of dry eye, new concepts of dysfunctional dry eye, advances its diagnosis and classification, refinement and standardization of dry eye treatment, and the future development of dry eye research.
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Objective@#To compare the effects of femtosecond laser-assisted phacoemusification and microincision phacoemusification on ocular surface.@*Methods@#A nonrandomized comparative cohort study was designed.The study recruited 116 eyes of 116 patients with age-related cataract in Hankou Aier Eye Hospital from January to March 2018.Consecutive patients were divided into femtosecond laser-assisted phacoemulsification group (femtosecond laser group for short) (55 eyes) and microincision phacoemulsification group (microincision group for short) (61 eyes). The ocular surface disease index(OSDI), break-up time of tear film (BUT), corneal flurescein staining, Schirmer Ⅰ test (SⅠt) and goblet cell density were sequentially evaluated preoperatively and postoperatively at 1 week and 1 month.This study followed the Declaration of Helsinki.Written informed consent was obtained from each subject prior to entering study cohort.This study protocol was approved by Ethic Committee of Hankou Aier Eye Hospital (No.2017IRB01).@*Results@#At 1 day after surgery, the best corrected distance visual acuity (BCDVA) (LogMAR visual acuity) was 0.14(0.10, 0.24) and 0.20 (0.17, 0.27), respectively in the femtosecond laser group and the microincision group.BCDVA in the femtosecond laser group was better than that in the microincision group, with a significant difference between them(U=-3.140, P=0.002). Compared with the microincision group, the operation time was longer, the phaco time was shorter, the cumulative dissipated energy was lower, and the balanced salt solution perfusion volume was less in the femtosecond laser group, with significant differences between them (all at P<0.001). Compared with the pre-operation, the mean OSDI socres, BUT and goblet cell density were significantly decreased, and the fluorescein staining scores were significantly increased at 7 days and 30 days after surgery in the femtosecond laser group and the microincision group, with significant differences between them (all at P<0.05). The SⅠt value fluctuations in the two groups were small at different time points before and after operation, and the overall comparative differences between the two groups and different time points had no statistical significances (Fgroup=0.510, P=0.477; Ftime=0.741, P=0.479).@*Conclusions@#Compared with microincision phacoemusification, patients who had femtosecond laser-assisted phacoemusification have more severe ocular surface staining, shorter tear film time and more reduction of goblet cell count at the early stage.
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Objective To evaluate the feature of meibomian glands and tear film lipid layer thickness (LLT) in dry eye of children with long-term visual display terminals (VDTs) exposure.Methods Cross-sectional case control observation was performed.Eighty-four 6-14 years old children (149 eyes) with long or short time VDTs exposure from January to August 2017 in Tianjin Medical University Eye Hospital were enrolled,including 29 children 56 eyes for long time VDTs group and 55 children 93 eyes for short time VDTs group.Questionnaires were designed to detect disease history and ocular surface disease index (OSDI).Slit lamp microscopy,Keratograph 5 M,lipiview ocular surface interometry examination,corneal-conjunctival fluorescence staining,tear film break-up time (BUT) and Schirmer test (S I t) were performed on all the subjects.The subjective OSDI and clinical signs were compared between long time VDTs group and short time VDTs group and correlation between OSDI and clinical signs were analyzed in each group.This study followed the declaration of Helsinki.Informed consent was obtained from each subject or the guardian prior to any medical examination.Results Compared with the short time VDTs group,high OSDI value and long average noninvasive BUT (NIBUTav) were observed in the long time VDTs group,the differences were statistically significant (OSDI:t =-2.662,P =0.009;NIBUTav:Z =2.315,P =0.021).The meibomian gland dropout score was significantly different between the 2 group (x2 =11.779,P =0.008).In long time VDTs group,OSDI was negatively correlated with S I t (rs =-0.598,P =0.050) and positively correlated with meibomian gland dropout score and NIBUTav (rs =0.522,P =0.022;rs =0.597,P =0.011).In short time VDTs group,OSDI was positively correlated with first time noninvasive BUT (NIBUTf),NITBUTav and partial blink rate (rs =0.372,P =0.030;rs =0.408,P =0.008;rs =0.426,P =0.050).Conclusions Compared with short time VDTs exposure,the value of subjective OSDI was high and meibomian gland dropout was serious in long term VDTs exposure dry eye children.The OSDI value is associated with NIBUTf and partial blink ratio in short time VDTs dry eye children.
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Objective To evaluate the effectiveness of intense pulsed light (IPL) therapy for meibomian gland dysfunction (MGD).Methods A prospective senal cases observational study was designed.Thirty patients with MGD (60 eyes)in the Tianjin Medical University Eye Hospital from October 2017 to April 2018 were enrolled.All the subjects received IPL therapy at baseline,1 week and 3 weeks after the baseline,respectively.Before each treatment and 3 months after treatment,subjective symptoms (ocular surface disease index [OSDI] questionnaire,Standard Patient Evaluation of Eye Dryness [SPEED] questionnaire),palpebral margin score,lipiview ocular surface interferometer lipid layer thickness (LLT) measurement,tear meniscus height (TMH) and noninvasive tear film breakup time (NIBUT) measurement were performed to detect and assess the IPL treatment effect.The safety of the IPL treatment was evaluated through the visual acuity and intraocular pressure.Results The OSDI score,SPEED score and the score of lipid excretion and secretion of meibomian gland before treatment were significantly higher than those of the other three time points (all at P＜0.01).The TMH before treatment was significantly lower than that of the other three time points (all at P＜0.01).No significant differences were obtained on first time NIBUT,blinking times and incomplete blink ratio among different time points (all at P＞0.05).After treatment,the average thickness of tear film lipid layer was slightly increased,and the secretion of tarsal gland was improved.There were no adverse reactions during the treatment and follow-up,no changes in corrected vision,and the intraocular pressure fluctuated within the normal range.Conclusions IPL therapy can alleviate the symptoms and signs of dry eye of MGD patients,and the effect can last for at least 3 months.IPL is an effective and safe method in treatment of MGD.
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Objective To investigate the inhibitory effect of bromfenac sodium hydrate ophthalmic solution on corneal neovascularization (CNV) induced by alkali burn.Methods A total of 192 specific pathogen free (SPF) degree adult male Sprague-Dawley (SD) rats were used in this study.One hundred and seventy-two rats were chosen to establish CNV model with alkali burn in the right eyes.Following alkali burn,rats were randomly divided into CNV group,model control group,bromfenac sodium group and fluorometholone group,with 43 rats (43 eyes) in each group.Another 20 rats (40 eyes) served as normal control group.One day after modeling,the model control group,bromfenac sodium group and fluorometholone group received phosphate buffer saline (PBS),bromfenac sodium hydrate ophthalmic solution and 0.1％ fluorometholone eye drops,respectively.The state of cornea and anterior chamber and the growth of CNV of rats in each group were observed by slit-lamp microscope every day after modeling.At 1,3,7,14,21 and 28 days after modeling,the anterior segment photos of the experimental eyes were captured,and the percent of cornea areas covered by CNV was calculated.At 7,14 and 28 days after modeling,the eye tissue sections were stained with hematoxylin and eosin staining and immunohistochemistry staining to evaluate the expressions of CD45 and VEGF-A.Real-time quantitative PCR and enzyme-linked immunosorbent assay (ELISA)were used to detect the expression of COX-2 and VEGF mRNA and protein level.The use and care of the animals complied with the Statement of the Association for Research in Vision and Ophthalmology(ARVO).Results Each model group showed corneal edema and opacification 1 day after modeling.The corneal edema was aggravated 7 days after modeling.On the 14th day after modeling,the degree of corneal opacity and edema decreased gradually.On the 28th day after modeling,leucoma was observed in CNV group and model control group,and nebula was observed in bromfenac sodium group and fluorometholone group.At 7,14,21 and 28 days after modeling,the percentages of CNV areas in bromfenac sodium group and fluorometholone group were significantly lower than those in CNV group and model control group (all at P＜0.05).No significant difference was found in the percentage of CNV areas between bromfenac sodium group and fluorometholone group at various time points (all at P＞0.05).On the 7th day after modeling,the thinning of corneal epithelial layer,edema and arrangement disorder of stroma layer were observed,and the expression of VEGF-A was positive in all model groups;a small amount of CD45 positive inflammatory cell infiltrations were observed in CNV group and model control group.On the 14th and 28th day after modeling,CNV was seen in the center of cornea in CNV group and model control group;the epithelial keratosis and reduction of corneal edema were seen in each group,and no inflammatory cell infiltration was observed in each group.On the 7th day after modeling,the expressions of COX-2 and VEGF mRNA in CNV group and model control group were significantly higher than those in normal control group,bromfenac sodium group and fluorometholone group (all at P ＜ 0.05),the expressions of COX-2 and VEGF protein in bromfenac sodium group were significantly lower than those in CNV group (all at P＜0.05).The corneal peroration rate in model control group and bromfenac sodium group was 10％ (1 case in 10 rats).The corneal perforation rate in fluorometholone group was 30％ (3 cases in 10 rats).In each model group,10％ to 30％ rats had hyphema.Conclusions Bromfenac sodium hydrate ophthalmic solution can inhibit the formation and growth of CNV after alkali burn in rats.This effect may be mediated by regulating COX-2 expression,reducing inflammation and inhibiting VEGF production.
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Objective To synthesize a dual-function nanodrug with immunosuppression of tacrolimus (FK506) and lubrication of artificial tear carboxymethylcellulose sodium (CMC), and provide a basis for the clinical treatment of dry eye syndrome. Methods The FK506 nanocrystals were prepared by ultrasonic method, and polyallylamine hydrochloride (PAH) and CMC were deposited alternatively on the surface of FK506 nanocrystals using layer-by-layer (LbL) self-assembly technology to prepare the bifunctional nanodrug. The morphology, particle size, surface charge, and composition of the nanodrug were analyzed. Results The particle size of FK506 nanocrystals was uniform, and the FK506-(PAH/CMC)3 was approximately spherical with uneven surface. Zeta potential detection results showed that the charge changed alternatively with the increasing of layer number. The results of laser scanning confocal imaging and infrared spectroscopy showed that PAH and CMC were successfully modified on the surface of FK506 nanocrystals. Conclusion The prepared FK506-(PAH/CMC)3 nanodrug is expected to provide a scientific basis for the combined treatment of dry eye.
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Objective To investigate the effect of mesenchymal stem cells (MSCs) combined with low-dose cyclosporin A ( CsA) on corneal transplantation rejection and its mechanism. Methods A fully allogeneic rat cornea transplant model (Wistar rats to Lewis rats) was created. Thirty-six model rats were divided into MSCs group, CsA group and MSCs+CsA group randomly by using the random number table method,with 12 rats for each group. In each group,six rats were used for the evalution of symptom observation and six for the assay of immune factor. Three days after corneal transplantation,slit lamp microscope was used to observe and record the vascularization,edema and opacity scores of corneal graft. After 18 days,spleen T helper cell(Th)1 and Th2 cytokines were measured by enzyme linked immunosorbent assay ( ELISA ). The use and care of the animals complied with Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. This study protocol was approved by Ethic Committee of Tianjin Medical University Eye Hospital (No. TMUaMEC2017028). Results The survival time of corneal grafts in MSCs+CsA group was prolonged in comparison with the MSCs group and CsA group,with significant differences between them([21.3±3.9]days vs. (14.8±2.4)days; [21.3±3.9]days vs. [16.0±1.1]days) (P=0.003,0.004). Eighteen days after corneal transplantation,corneal opacity scores were 3.17±0.17, 3.00±0.00 and 2.17±0.17,corneal neovascularization scores were 2.67 ± 0.21,2.33 ± 0.21 and 1.83 ± 0.21, respectively in the MSCs group, CsA group and MSCs+ CsA group, showing significant differences among the three groups (F=15.500,P<0.01; F=4.524,P=0.029). The corneal opacity scores and corneal neovascularization scores in the MSCs+ CsA group were significantly lower than those in the MSCs group and CsA group(all at P<0.05). Interleukin(IL)-2,IL-4,IL-10,and interferon-γ (IFN-γ) concentrations were all significantly different among the MSCs group,CsA group and MSCs+ CsA group (F=15.000,12.810,10.720,17.960,all at P< 0.01). Compared with the MSCs group and CsA group,Th1 subsets cytokines IFN-γ and IL-2 concentrations in the MSCs+CsA group were significantly lower,and Th2 subsets cytokines IL-4 and IL-10 concentrations were significantly increased in the MSCs+CsA group in comparison with the MSCs group and CsA group ( all at P<0.05 ). Conclusions The combination of MSCs with low-dose CsA can effectively ameliorate immunological rejection by the modulation of the balance of Th1/Th2.
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Objective To compare the pharmacodynamics between different batches of recombinant decoy receptor innovative drug RC28-E1 and RC28-E2 in retinal angiogenesis and neovascularization,and analyze its mechanism.Methods Sixty postnatal Day 4 (P4) C57BL/6J mice were randomly divided into normal control group,vascular endothelial growth factor (VEGF)+fibroblast growth factor 2 (FGF2) group,VEGF+FGF2+RC28-E1 group,VEGF+FGF2+RC28-E2 group,VEGF+FGF2+conbercept group and VEGF+FGF2+FGF trap group by using a random number table,with 10 mice in each group.The mouse retinal explant culture system was established,and stimulated with the corresponding factors and drugs prepared in the starving culture media.The normal controls were treated with the starving media.Then the retinal explants were stained with Isolectin B4 and imaged.The number of filopodia per vascular length was quantified.In addition,ninety-six P7 C57BL/6J mice were randomly divided into normal control group,oxygen-induced retinopathy (OIR) model control group,OIR + RC28-E1 group,OIR + RC28-E2 group,OIR+conbercept group and OIR+FGF trap group by using a random number table,with 16 mice in each group.The normal controls were raised under normoxia for 10 days,and the rest of the groups were raised under hyperoxia for 5 days,then returned to normoxia for another 5 days.On P17,the retinas were isolated and stained with Isolectin B4.The stained retinas were mountedon the slides and photographed.The relative vessel obliteration and neovascularization in retina were analyzed with computer software.Then the protein levels of VEGF and FGF2 were examined by Western blot in the retinas of each group in the OIR experiment.Finally,in the RF/6A cells stimulated with VEGF and FGF2,the activities of the signaling pathways,including MEK-extracellular regulated protein kinases (Erk),protein kinase C (PKC) and protein kinase B (Akt) pathways,were examined by Western blot.All experimental procedures were evaluated and approved by the Institutional Animal Care and Use Committee of Tianjin Medical University (SYXK 2009-0001),and were in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.Results The results of retinal explant cultures showed that the numbers of filopodia per vascular length in VEGF + FGF2 + RC28-E1,VEGF + FGF2 + RC28-E2,VEGF + FGF2 + conbcrcept,and VEGF+FGF2+FGF trap groups were all significantly less than that in the VEGF+FGF2 group (all at P ＜ 0.001).The filopodia number in retinal vascular front in RC28-E1 group was similar to that in the RC28-E2 group (P =0.15),whereas the filopodia numbers in both groups were significantly decreased as compared to those in VEGF+ FGF2+conbercept group and VEGF+FGF2+FGF trap group (all at P＜0.001).The results from the OIR mouse model showed that the relative vessel obliteration area in OIR model control group was dramatically higher than those in the drug intervention groups (all at P＜0.05).There was no statistical significance in the relative vessel obliteration area between OIR+RC28-E1 group and OIR+RC28-E2 group (P =0.17),while the obliteration areas in both RC28-E-intervened groups were significantly lower than those in the OIR+conbercept group and OIR+FGF trap group (all at P＜0.05).The relative neovascular pixels in the intervention groups were significantly lower than those in the OIR model control group (all at P＜0.001).The neovascular pixels in OIR+RC28-E1 group were significantly lower than those in VEGF+FGF2+conbercept group and VEGF + FGF2 + FGF trap group (both at P ＜ 0.05),but comparable to those in OIR+RC28-E2 group (P =0.39).Western blot result showed that,the protein expression of VEGF and FGF2 in the OIR mouse retinas were significantly upregulated compared to those in the normal ones (both at P＜0.001).The upregulation of both genes were normalized by both RC28-E1 and RC28-E2.In addition,the stimulation of VEGF and FGF2 induced an enhanced activity in MEK-Erk pathway in RF/6A cells,whereas RC28-E1 inhibited the overactivation.Conclusions RC28-E1 and RC28-E2 both can inhibit angiogenesis in the retinal explants isolated from neonatal mice;they also reduce vessel obliteration and mitigate neovascularization in the OIR mouse model.Therefore,the pharmacology batch and pilot test batch of RC28-E have similar efficacies and reliable stability,and are superior in the anti-angiogenic and anti-neovascular efficacy to the currently clinically available drugs conbercept and FGF trap.RC28-E1 may suppress pathological neovascularization through inhibiting the overactivation of MEK-Erk pathway in retinal vascular endothelial cells.
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Objective This study was to study the effect of 0.1％ and 0.3％ sodium hyaluronic ophthalmic solution on tear film stability and visual quality of dry eye patients after femtosecond laser-assisted laser in situ keratomileusis (FS-LASIK).Methods A prospective randomly controlled study was designed.Pre-surgery normal patients were evaluated again at 1 week after FS-LASIK,60 mild and severe dry eye patients were enrolled from April 2016 to April 2017 in Tianjin Medical University Eye Hospital.Then 0.1％ and 0.3％ sodium hyaluronate were randomly given in each dry eye group.Ocular surface disease index (OSDI),optical quality analysis system Ⅱ(OQAS Ⅱ),Keratography 5M and corneal staining were performed before operation,1 week (before instillation and 15,30,60,120 and 180 minutes after instillation),1 month and 3 months after operation.This study was approved by the Ethic Committee of Tianjin Medical University Eye Hospital (2015KY-04).Informed consent was obtained from all subjects after explanation of the nature and possible consequence of the study.Results The 0.3％ sodium hyaluronate group was better than 0.1％ sodium hyaluronate group in alleviating OSDI score and cornea staining at 3 months in the mild dry eye group (Fgroup =10.913,P =0.003;P =0.027).Compared with the before operation,the objective scatter index(OSI) was increased at 1 week before instillation and many different time points after instillation in both 0.1％ sodium hyaluronate group and 0.3％ sodium hyaluronate group,with significant differences between them (all at P＜0.05),there was no statistically significant difference in OSI at 1 month and 3 months after surgery compared with before surgery in both 0.1％ and 0.3％ sodium hyaluronatc group.In 0.1％ sodium hyaluronic group and 0.3％ sodium hyaluronic group,the mean value of total OSI at 1 week after surgery were siginifcantly higher than that before surgery (P=0.046,0.060).In 0.1％ sodium hyaluronate group,the mean values of total OSI at various time points after instillation were not statistically different from that before surgery (all at P＞0.05),total OSI at 1 month and 3 months after operation were higher than that before surgery (P =0.047,0.017);In 0.3％ sodium hyaluronic group,the mean value of total OSI at 15,30,60 and 120 minutes after instillation were significantly higher than that before surgery (all at P＜0.05),and compared with the total OSI before surgery,no statistical differences were found at 1 month and 3 months after operation (P =0.046,0.667).Significant differences were found in NIBUT avg among different time points in the mild dry eye patients (Ftime =2.290,P =0.022).In 0.1％ sodium hyaluronic group and 0.3％ sodium hyaluronic group,the NIBUT avg at 1 week after surgery were significantly lower than that before operation (both at P＜0.05).Compared with the before operation,no significant differences were found at the following 3 hours after instillation in 0.1％ sodium hyaluronic group (all at P＞0.05).In 0.3％ sodium hyaluronic group,the NIBUT avg at 60,120 and 180 minutes after instillation were significantly lower than that before surgery (P=0.040,0.047,0.041),and NIBUT avg at 3 months after operation was significantly lower than that before surgery (P =0.044).Conclusions The 0.3％ sodium hyaluronic is better than 0.1％ sodium hyaluronic in alleviating the OSDI score and cornea staining on the mild dry eye post FS-LASIK,and the tear film stability and related visual quality fluctuated over time after the application of 0.3％ sodium hyaluronic.
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Objective To analyze the correlation between the dry eye symptom and signs in middle-aged and aged people by Lipiview(R) ocular surface interferometry.Methods A total of 170 eyes from 85 middle-aged and aged people (55-80 years old) were examined with Standard Patient Evaluaion of Eye Dryness(SPEED) questionnaire and Lipiview(R) ocular surface interferometry,meanwhile obtaining the consent of the subjects.Then the correlation between the symptom and Lipid Layer Thinckness (LLT) and partial blink rate were analyzed.Results SPEED questionnaire score was 6.13 ±4.60.LLT was(74.88 ± 21.16) nm.LLT variability was 4.47 ± 3.40.The partial blink rate was 0.62 ± 0.36.The Spearman correlation analysis indicated that the nega-tive correlation was shown between the SPEED questionnaire score and LLT (r =-0.823,P =0.000),SPEED questionnaire score and LLT variability(r =-0.268,P =0.018).The positive correlation were found between LLT and Std.Dev of LLT (r =0.339,P =0.030).However,the results of the SPEED score were uncorrelated with partial blink rate (P ＞ 0.05).Conclusion The results from this study demonstrate that correlation among symptom,LLT and LLT variability,but poor correlation between symptom and partial blink rate.
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Background Reasearches showed that α-melanocyte-stimulating hormone (α-MSH) inhibits inflammation and ameliorates the ocular surface abnormalities in a scopolamine-induced dry eye rat model,and the managing effect of sodium carboxymethylcellulose (CMC) on dry eyes also has been determined.However,whether α-MSH can enhance the therapeutic effects of CMC remains to be investigated.Objective This study was to investigate the protective effects of α-MSH combined with CMC on ocular surface in a scopolamine-induced dry eye rat model.Methods Sixty clean female Wistar rats were randomly divided into normal control group,model control group,NaCl group,CMC group,α-MSH group and α-MSH+CMC group,and 10 rats for each group.The dry eye models were established by subcutaneous injection of scopolamine hydrobromide at 9:00,12:00,15:00 and 18:00 per day for 28 days.0.9％ NaCl solution,1×10 3 mg/ml α-MSH solution,0.5％ CMC eye drop,and 1 ×10-3 mg/ml α-MSH+0.5％ CMC solution were topically administered twice a day (8:00,17:00) since the initial day of modeling according to grouping.Shirmer Ⅰ test (S Ⅰ t),breakup time of tear film (BUT) and corneal fluorescence staining were performed before and 7,14,21,28 days after the application of drugs.At 28 days following the administration of drugs,the eyeballs of the rats were collected.Hemotoxylin and eosin staining was employed to examine the morphology of corneas,and periodic acid schiff (PAS) staining was used to count the conjunctival goblet cells.This study protocol was approved by Experimental Animal Ethic Committee of Tianjin Medical University (SYXK 2009-0001),and the use and care of the rats complied with ARVO Statement.Results The S Ⅰ t and BUT values were significantly reduced,and the corneal fluorescence staining scores were significantly increased over time following modeling in the model control group (all at P＜0.01).No significant differences were found in the S Ⅰ t,BUT and corneal fluorescence staining scores between model control group and NaCl group at various time points (all at P＞0.05).At 7,14 and 21 days after intervention,the S Ⅰ t values were (4.800±0.789),(4.100±0.516) and (4.300±0.856) mm in the α-MSH+CMC group,which were considerably higher than (2.875 ±0.719),(2.375 ±0.619) and (2.532±0.957)mm in the NaCl group (all at P＜0.01).At 7 days after intervention,the BUT values were (4.938± 1.843) seconds and (5.000±1.491) seconds in the α-MSH group and α-MSH+CMC group,which were significantly higher than (3.250±1.000) seconds in the NaCl group (both at P＜0.01).The corneal fluorescence staining scores in the CMC group,α-MSH group and α-MSH+CMC group were significantly lower than that in the NaCl group,with the lowest score in the α-MSH +CMC group (all at P＜0.05).The thickening of corneal epithelial layer,corneal edema and arrangement disorder of corneal stroma were found in the model control group and NaCl group;while slight corneal edema and epithelial cell proliferation were exhibited in the α-MSH+CMC group by hemotoxylin and eosin staining.PAS staining showed that the number of goblet cells was much more in the CMC group,α-MSH group and α-MSH+ CMC group than that in the model control group and NaCl group (all at P ＜ 0.01).Conclusions The sole application of α-MSH or CMC alleviates ocular surface damage and morphological abnormality to certain extent,and the combination of α-MSH and CMC generates more effective protection in comparison with sole administration of α-MSH or CMC.The early application of the drugs plays an improvement role in tear secretion and tear film stability in dry eyes.
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Background Obstructive meibomian gland dysfunction (OMGD) is due to the meibomian gland duct obstruction and/or meibum abnormal secretion.Meibomian gland morphological change is important to clinical observation.Objective This study aimed to image the meibomian gland structures and the morphological changes in eyes with OMGD using a non-contact infrared meibography system (Keratograph 5M).Methods A case-control study was performed in Tianjin Medical University Eye Hospital from January to June in 2015.Sixty OMGD patients and 60 age-matched healthy controls were recruited.Ocular Surface Disease Index (OSDI) questionnaire was employed for the assess of ocular surface symptoms,and slit-lamp examination was performed for the observation of lid margin and meibomian gland orifices.Meibomian gland secretion scores and Keratograph 5M examination the morphological changes of the meibomian glands.According to OSDI questionnaire,the patients of OMGD were classified into mild group (12.0 ＜ OSDI ≤ 22.0) (23 eyes),moderate group (22.0 ＜ OSDI ≤ 32.0) (21 eyes) and severe group (32.0 ＜ OSDI ≤ 100.0) (16 eyes).Partial or complete loss of the meibomian glands (meibomian dropout) was scored from grade 0 (no loss) through out grade 3 (lost area was ＞2/3 of the total meibomian gland area).Written informed consent was obtained from all subjects before the examination.Results The OSDI,meibomian gland orifices,meibomian gland secretion scores and meibomian gland dropout scores were significantly higher,and BUT was dramatically shorter in the OMGD group than those in the normal group,showing significant differences between the two groups (Z =-9.425,-8.954,-7.781,all at P＜0.05).The BUT was (6.10± 1.91) s and (10.67-±2.11)s,and meibomian gland dropout scores were 1.35±0.90 and 0.88±0.64 in the OMGD group and normal control group,showing significant differences between the two groups (t=3.276,-12.418,both at P＜0.05).OSDI,meibomian gland secretion scores and meibomian gland dropout scores were significantly in three intergroups (H=52.121,13.642,37.180,all at P＜0.05).BUT,meibomian gland orifices were significantly different among the mild,moderate and severe OMGD (F=58.191,8.474,both at P＜0.05).OSDI showed a significantly high positive correlation with meibomian gland orifices,meibomian gland secretion scores and meibomian gland dropout scores (r =0.708,P =0.000;r =0.759,P =0.000;r =0.270,P =0.003).BUT showed negative correlation with meibomian parameters (r=-0.692,P=0.000;r=-0.691,P=0.000;r=-0.192,P=0.036).Conclusions Keratograph 5M provides an effective noninvasive method in assessing the image of the meibomian gland structures and meibomian gland dropout.There are differences in the ocular surface and meibomian glands between the OMGD patients and normal persons.
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Objective To assess the optical quality after implantation of the intraocular contact lens with and without a central hole for high myopia.Methods A total of 23 patients (41eyes),including 11 patients (21 eyes) implanted with V4 ICL (V4 ICL group) and 12 patients (20 eyes) with V4 c ICL implantation (V4 c ICL group) were enrolled in this research.Six months after operation,slit lamp microscope was used to check the anterior segment,and LogMAR chart was to test uncorrected visual acuity (UCVA) and best corrected visual acuity (BCVA).Moreover,automatic refractometer was to measure the spherical equivalent (SE) and astigmatic diopter before and after the operation.OQAS parameters included objective scattering index (OSI),modulation transfer function (MTF),Stryrby (SR),OQAS values (OV100％,OV20％,OV9％) at different contrast level were examined.Results In both groups,the parameters of UCVA (logMAR),BCVA (logMAR),SE and astigmatic diopter at 6 months after operation were significantly improved compared with before operation,with significant difference (all P ＜ 0.05).There was no siguificant difference in the UCVA,BCVA,SE and astigmatic diopter between the two groups before operation and 6 months after operation (all P ＞ 0.05).Moreover,the values of MTFcutoff,SR,and OQAS visual parameters (OV100％,OV20％ and OV9％) at 6 months after operation were significantly higher than those before operation between the two groups,and the difference was statistically significant (all P ＜ 0.05),while OSI was lower than preoperative data,but there was no significant difference in OSI between the two groups (P ＜ 0.05).There was no significant difference in OQAS visual quality parameters,including OSI,MTFcutoff,SR,and OV100％,OV20％ and OV9％,between the two groups (all P ＞ 0.05).Conclusion It is safe and effective of the two types of intraocular contact lens implantation for correction of high myopia,and V4 c ICL can achieve the similar visual quality to the traditional V4 ICL.
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Background Corneal transplantation is a primary method for the treatment of serious corneal diseases, but its application is limited because of the shortage of corneal donor.The study on tissue engineering corneal epithelium provides a new approach to corneal transplantation, and the biological scaffold materials for tissue engineering corneal epithelium is an issue of increasing concern.Bacterial cellulose membrane has been used in medical field,but its application in tissue engineering corneal epithelium deserves more researching.Objective This study was to evaluate the biocompatibility of bacterial cellulose membrane as a biological scaffold of tissue engineering corneal epithelium.Methods Corneal epithelium was isolated from 1 month-old New Zealand White rabbit.Corneal epithelial cells were cultured using explant method and identified by detecting the CK-3 expression using immunofluorescence technique.The second generation ceils were inoculated on bacterial cellulose membrane and culture plate, respectively, and the growth status of the cells were examined and compared under the optical microscope.The cell activity/toxicity test was performed by LIVE/DEAD cell staining kit at the third day after inoculation to evaluate the survival rate.The ultrastructure of the cell surface was examined under the scanning electron microscope.The study was performed in accordance with the ARVO Statement.Results Rabbit corneal epithelial cells grew well 1 week after primarily cultured with a cobblestone-like appearance and positive response for CK3 antibody.The cells on the bacterial cellulose membrane presented a round shape and regular arrangement and showed the green fluorescence for LIVE/DEAD test,with the survival rate 100％.Abundant leafy protrusion, microvilli and intercellular junction were seen under the scanning electron microscope.In addition, mitosis phase of cells and many filopodia between the cells and bacterial cellulose membrane were also exhibited.Conclusions Rabbit corneal epithelial cells can grow well in bacterial cellulose membrane.Bacterial cellulose membrane has good biocompatibility, indicating that bacterial cellulose membrane can be used as new biological material for tissue engineering corneal epithelium.