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AIM: To explore the dynamic expression of high mobility group box 1(HMGB1)in scar tissues after glaucoma drainage valve implantation, and to further reveal the role and possible mechanism of HMGB1 in scarring after glaucoma surgery.METHODS: A total of 60 New Zealand white rabbits were randomly divided into control group(n=20), model group(n=20, silicone implantation under conjunctival sac)and model with drug administration group(n=20, silicone implantation under conjunctival sac combined with 5-fluorouracil injection). The conjunctival tissues were collected at 4 and 8 wk after surgery. HE staining and Masson staining were used to detect the proliferation and distribution of fibroblasts and collagen fibers in conjunctival tissues. Immunohistochemistry was utilized to detect the distribution and changes of HMGB1, transforming growth factor(TGF)-β1, Smad3 and α-smooth muscle actin(SMA)in conjunctival tissues. RT-PCR and Western blot were adopted to detect the mRNA and protein expression of HMGB1, TGF-β1, Smad3 and α-SMA in conjunctival tissues.RESULTS: HE staining and Masson staining showed that the proliferation of inflammatory cells, fibroblasts and collagen fibers in the model group was significantly higher than that in the control group at both 4 and 8 wk. Meanwhile, the proliferation of fibroblasts and collagen fibers in the model with drug administration group was significantly lower than that in the model group. Immunohistochemical staining showed that the expression of HMGB1, TGF-β1, Smad3 and α-SMA protein was observed in the conjunctival tissues of the model group both 4 and 8 wk, with brown and significantly deeper staining of the model group at 8 wk. Meanwhile, the positive staining in the model with drug administration group at both 4 and 8 wk was significantly lower than that in the model group. There was positive correlations between the number of fibroblasts stained with HE and the expression of HMGB1 in the conjunctival tissue of the model group at both 4 and 8 wk(r=0.602, 0.703, all P<0.05). RT-PCR and Western blot revealed that the mRNA and protein expression levels of HMGB1, TGF-β1, Smad3 and α-SMA in the model group were significantly higher than those in the control group at both 4 and 8 wk(all P<0.05). Meanwhile, the mRNA and protein expression levels of HMGB1, TGF-β1, Smad3 and α-SMA in the model with drug administration group were significantly lower than those in the model group(all P<0.05). There was positive correlations between mRNA expressions of HMGB1 and TGF-β1, Smad3 in the model group and the model with drug administration group(all P<0.05).CONCLUSION: The expression of HMGB1 increased at a time-dependent manner after glaucoma valve implantation. HMGB1 acts an indispensable role in the initiation and progression of scar formation after glaucoma surgery, which may be involved in the regulation of TGF-β/Smad signaling pathway.
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Objective To explore the value of qualitative pattern of isolated-check visual evoked potential ( ic-VEP) in the early diagnosis of primary open-angle glaucoma ( POAG) , and to provide an objective and effective basis for the early diagnosis of clinical glaucoma. Methods Sixty-eight patients (82 eyes) with normal POAG were randomly selected. Forty-seven normal individuals (76 eyes) were as control group. Both groups were treated to analyze ic-VEP,the defect of the visual field, the thickness of the retinal nerve fiber layer ( RNFLT) , and the mult-ivariate correlation analysis between ic-VEP, the visual field and RNFLT was performed. The sensitivity and speci-ficity of ic-VEP in the diagnosis of POAG was analysis in this study. Results There were no significant differences in age, diopter and central corneal thickness between glaucoma group and normal control group. However, the in-traocular pressure ( IOP) , the visual field, RNFLT and ic-VEP between two groups were statistically different( P＜0.01) . The correlation analysis of ic-VEP Signal-to-noise ratio ( SNR) with diopter, IOP, the visual field and RN-FLT in the glaucoma group showed that ic-VEP SNR was significantly associated with IOP, the visual field and RN-FLT damage(P＜0.01). The sensitivity of ic-VEP to detect POAG was 90.2% and the specificity was 88.2% . Ic-VEP and the visual field, RNFLT had good consistency. Conclusion Ic-VEP has high sensitivity and accuracy in the diagnosis of primary open-angle glaucoma. Meanwhile ic-VEP SNR is associated with the damage of retinal nerve fiber layer and the visual field damage.
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Objective To research the genetic susceptibility of proliferative diabetic retinopathy (PDR) in Han patients with type 2 diabetes from Southern China.Methods A cross-sectional study was performed under the informed consent of the patients.Patients with type 2 diabetes in the Dongguan Eye Study from September 2011 to February 2012 and relative patients treated in Guangdong General Hospital from July 2017 to March 2018 were included in this study,including 100 patients with diabetes mellitus(DM) and 120 patients with PDR.Whole exome sequencing was used to identify DNA mutation in peripheral blood samples from 22 type 2 diabetic patients without retinopathy (DM group) and 23 diabetic patients with PDR (PDR group).Genotype and allele of the nine selected single-nucleotide polymorphisms (SNPs) were tested and analyzed by SnaPshot technology in another 78 DM patients without retinopathy and 97 PDR patients.Results A total of 75 SNPs were associated with PDR (P＜0.01),involving 53 genes.Eleven gene loci were in the exon region and 7 were non-synonymous mutations.Nine exon loci of 8 genes with significant differences were screened out for the verification.SnaPshot SNP genotyping technique found that there were no significant differences in allele and genotype frequency in the nine selected SNPs between PDR group and DM group (all at P＞0.05).However,7 haplotypes distribution frequencies were significantly different between PDR group and DM group (all at P＜0.01).Hapl and Hap4 might reduce the risk of PDR (both at OR＜1,P＜0.05),and Hap2 might increase the risk of PDR (OR＞ 1,P＜0.05).Conclusions The occurrence of PDR probably has a genetic susceptibility in type 2 DM patients of Han nationality in Southern China.
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Background Experimental autoimmune uveitis (EAU) is a common animal model of uveitis.Natural killer (NK) cells have been confirmed to be a type of strong inflammation-causing cells,but its role in EAU is still studing.Objective This study was designed to explore the role and mechanism of NK cells in the pathogenesis of EAU.Methods Thirty-six SPF Lewis rats were randomly divided into expeimental control group and EAU 6-,9-,12-,16-,and 21-day groups (6 rats for each group).Rats in EAU group received subcutaneous injection interphotoreceptor retinoid binding protein (IRBP) combining 5 mg/ml tubercle bacillus with complete Freund's adjuvant (CFA) emulsion in foot pads,and then 400 ng pertussis toxin was intraperitoneally injected to extablish EAU models in the EAU 6-,9-,12-,16-,and 21-day group,and normal saline solution combined with CFA and 400 ng pertussis toxin was used in the same way in the experimental control group.The inflammatory response was observed by slit lamp daily after modeling and scored based on Caspi criteria.The eyeballs were extracted in 6,9,12,16 and 21 days after modeling for retinal histopathological examination,Immunofluorescent double-staining was employed to detect and locate the expression of NK cells in the retina.In addition,25 model rats were divided into EAU 0-,3-,6-,9-and 12-day groups,with 5 rats for each group,and eyeballs were extracted to prepare tissue homogenate.The expression of CXCL10 mRNA,and CXCL12 mRNA NK cell chemokines,in the tissue homogenate was assayed by real-time quantitative PCR.The use and care of the rats followed Regulations for the Administration of Affair Concerning Experimental Animal by State Science and Technology Commission.Results No inflammatory sign in ocular anterior segment of the rats was seen in the experimental control group.The expansion of rat iris vessels was found in the EAU 6-day group,and exudes and hypopyon of the anterior chamber occurred in the EAU 9-day group and the inflammation peaked in the EAU 12-day gorup.The rat retinal structure was normal in the experimental control group,and the arrangement disorder of retinal structure,the cell separation in outer nuclear layer and damage of photoreceptors were found under the optical microscope in different degree in various EAU groups,with the most serious change in the EAU 12-day group.Immunofluorescent double staining showed normally arranged nucleus in the experimental control group,and a lot of NK infiltration was seen in the EAU 6-day group and peaked in the EAU 9-day group.The expression level of CXCL10 mRNA in the EAU 9-day group was 34.298 ± 16.689,which was significantly higher than that in the EAU 3-,6-and 12-day group,respectively (1.390 ± 0.660,3.359 ± 2.581,4.711 ±1.387) (all at P＜0.01).No significant differences were found in the relative expression of CXCL12 mRNA among different EAU groups (F=2.851,P＞0.05).Conclusions Retinal NK cell infiltration occurs in the early stage of EAU,and the severity of NK cell infiltration is consistent with the inflammatory process and CXCL10 expression,suggesting NK cells play an important role in the early stage of EAU,and CXCL10 is an important chemokine of NK cells in EAU rats.