Age-related changes of corneal endothelial cell in healthy Chinese tree shrew measured by non-contact specular microscope

AIM：To determine the impact of age on the morphology of endothelial cells and central corneal thickness（CCT） in Chinese tree shrew.METHODS：One-hundred and twenty eyes of 60 healthy Chinese tree shrews were studied.Based on age,the tree shrews were divided into four groups.After general anesthesia,the images of endothelium were acquired using non-contact specular microscope Topcon 3000 P.Eight parameters of corneal endothelial cells were measured by built-in software,including CCT,endothelial cell density（ECD）,percent hexagonality（HG%）,coeffi cient of variability（CV）,size of minimal cell（Smin）,size of maximal cell（Smax）,average cells size（Savg） and size standard deviation（Ssd）.Data were analyzed using STATA software.The differences of eight parameters among groups and correlations with age were analyzed.RESULTS：In all studied animals,the average CCT was 2 4 9.6 ± 2 .29 μ m（202-301 μm）,ECD was 308 0.72 ± 460.76 cells/mm^2（1239.6-4047.6 cells/mm^2） and CV was 29.10±7.60（13.6-54.6）.CV was significantly different among different groups（P〈0.001）.Strong correlation with age was found in ECD,Smax,Savg,Ssd and CV.CONCLUSION：Cornea of Chinese tree shrews had half CCT of human cornea and similar ECD,CV and size of corneal endothelial cells.Young adult tree shrews had higher ECD,HG% and low CV.ECD,Smax,Savg,Ssd and CV correlated with age signifi cantly.

Ragweed pollen induces allergic conjunctivitis immune tolerance in mice via regulation of the NF-κB signal pathway

AIM:To investigate the feasibility and mechanism of immune tolerance in allergic conjunctivitis.METHODS:The allergic conjunctivitis immune tolerance mice model was established by ragweed pollen(RW)and the related cytokines were detected.The mice were divided into 9 groups and the maslinic acid(MA)or PBS were given for different group after modeling.The expression levels of chemokine ligand 5(CCL5)and P-65 in the conjunctival tissue were analyzed by immunohistochemistry,quantitative reverse transcription polymerase chain reaction(q RT-PCR)and Western blot.The percentage of interleukin-17(IL-17)and CD4+CD25+in the splenocyte supernatant was analyzed by flow cytometry.Fur thermore,the serum and splenocyte supernatant concentration of total-IgE,interleukin-10(IL-10),and IL-17 was analyzed by enzyme linked immune response(ELISA).RESULTS:After the model was established,symptoms of conjunctivitis were alleviated,the level of P-65,CCL5,IL-17,and total-IgE was raised,while the expression of IL-10,CD4+CD25+was decreased.This result fully demonstrated that a typical IL-17/regulatory-T-cells(Treg cells)imbalance and NF-κB activation.When the NF-κB signal pathway was suppressed,it showed that there was a further relief of conjunctivitis in mice.At the same time,the expression of total-IgE,IL-17,and CCL5 was decreased and the expression of anti-inflammatory factor(IL-10,CD4+CD25+)was increased.CONCLUSION:In the state of immune tolerance,symptoms of conjunctivitis in mice are alleviated,the Th-17 cells of allergic conjunctivitis mice are inhibited,and Treg cells activity is enhanced.

原位敲除猕猴视网膜RHO基因构建非人灵长类视网膜色素变性模型

遗传视网膜退行性疾病是全球范围内最主要的致盲疾病.该疾病主要的特征是视网膜感光细胞退化导致了不可逆的视觉能力丧失.视网膜色素变性(Retinitis pigmentosa,RP)便是其中的一大亚类,目前临床上尚无有效的治疗手段.本研究首次利用腺相关病毒(Adeno Associated Virus,AAV)介导的CRISPR/SaCa9技术,成功实现原位敲除目的基因RHO,建立了非人灵长类RP模型.研究结果发现,RHO敲除的猕猴视网膜表现出典型的RP感光细胞变性,视杆和视锥细胞渐行性死亡.在分子水平上,RHO基因成功地被敲除.RHO蛋白表达量也大幅度下降.免疫组化和电镜结果揭示了突变视杆细胞外段缺失或者变短,再现了RP最典型的特征.生理功能检测发现,RHO敲除视网膜的视网膜电流图(Electroretinogram,ERG)反应大幅度下降,证明了其视觉能力的受损.该动物模型为研究人类RP的发病机理以及制定相应治疗策略提供了重要的类人模型.