目的：利用生物信息学方法分析与葡萄膜恶性黑色素瘤转移相关的非编码RNA，以及它们作为竞争性内源RNA的作用机制。方法：从癌症基因组图谱(The Cancer Genome Atlas，TCGA)数据库下载80例葡萄膜恶性黑色素瘤患者的RNA测序数据和临床资料，采用edgeR算法分析转移与非转移患者组织中差异表达(differentially expressed，DE)的长链非编码RNA(lncRNA)、微小RNA(miR)和mRNA，并构建lncRNA-miR-mRNA的竞争性内源RNA(competing endogenous RNA，ceRNA)调控网络，基因富集分析和通路分析研究网络中mRNA的生物学功能。Kaplan-Meier生存曲线分析ceRNA网络中核心RNA与生存率的关系。结果：从发生远处转移的葡萄膜恶性黑色素瘤样本中，共鉴定出346个上调的mRNA，118个下调的miR和45个上调的lncRNA。其中67个mRNA，7个miR和30个lncRNA相互组合形成616个ceRNA单元，并形成了一个具有181条边线ceRNA网络。基因富集分析表明：网络中的mRNA富集在肿瘤生成和转移相关的几个基因本体(Gene Ontology)和信号通路。拓扑分析确定了6个核心lncRNA(LINC00861、LINC02421、BHLHE40-AS1、LINC01252、LINC00513和LINC02389)和3个核心mRNA(UNC5D、BCL11B和MTDH)。 所有核心lncRNA、核心mRNA的表达水平和5个miR(miR-221、miR-222、miR-506、miR-507、miR-876)的表达水平均与总体生存率显着相关(均P<0.05)。结论：本研究揭示了几种lncRNA及其相关的ceRNA网络在葡萄膜恶性黑色素瘤转移中的作用，为进一步研究葡萄膜恶性黑色素瘤的发生和/或转移提供了新的方向。

Objective: To elucidate the expression of long non-coding RNAs (lncRNAs) and their roles as competing endogenous RNAs (ceRNAs) in uveal melanoma (UM) metastasis. Methods: RNA sequencing data and clinical information of 80 patients with UM were obtained from The Cancer Genome Atlas (TCGA) database. Differentially expressed (DE) mRNAs, microRNAs (miR), and lncRNAs between metastatic and non-metastatic individuals with UM were screened using the edgeR algorithm. Gene enrichment analysis was conducted for the DE mRNAs. LncRNA-miR-mRNA regulatory triples and a ceRNA network were constructed. Betweenness centrality was used to screen hub genes and lncRNAs for subnetwork analysis. Kaplan-Meier survival analysis was conducted to explore correlations between the expression of hub RNAs and overall survival in the TCGA UM cohort. Results: A total of 346 upregulated mRNAs, 118 downregulated miRs, and 45 upregulated lncRNAs were identified in samples with systemic metastasis. Among them, 67 mRNAs, 7 miRs, and 30 lncRNAs mapped to 616 ceRNA triples, thus forming an interconnected ceRNA network with 181 edges. Gene enrichment analysis revealed that mRNAs in the network were enriched in multiple gene ontology terms and pathways associated with carcinogenesis and metastasis. Topological analysis identified 6 hub lncRNAs (LINC00861, LINC02421, BHLHE40-AS1, LINC01252, LINC00513, and LINC02389) and 3 hub mRNAs (UNC5D, BCL11B, and MTDH). The expression levels of all hub genes and 5 DEmiRs (miR-221, miR-222, miR-506, miR-507, miR-876) were significantly associated with the overall survival probability. Conclusion: This bioinformatic study revealed the functions of several lncRNAs and their associated ceRNA network in UM metastasis. It provides a novel in silicon evidence for future experimental study on the pathogenesis of systemic metastasis in uveal melanoma, especially from the perspective of non-coding RNA.

目的：评估外用白介素(Interleukin, IL)-6特异性抑制剂托珠单抗滴眼液在调控角膜碱烧伤后修复的安全性和有效性。方法：6只角膜假烧伤小鼠局部使用托珠单抗滴眼(2.5 mg/mL)和6只角膜假烧伤小鼠局部使用生理盐水滴眼，分别作为实验组和空白组以评估托珠单抗滴眼液的安全性。30只碱烧伤小鼠，按照1∶1随机分配到治疗组和对照组，治疗组使用托珠单抗滴眼液滴眼，对照组使用生理盐水，每日6次，连续用14 d。通过前段光学相干断层扫描观察虹膜前粘连、角膜后弹力层脱离及角膜水肿，在体视显微镜下检查角膜瘢痕形成及上皮伤口愈合。在角膜切片上评估IL-6定位、肌成纤维细胞、免疫细胞浸润和角膜上皮化生。在角膜铺片上评估角膜新生血管和新生淋巴管面积。通过实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction, qRT-PCR)方法检测小鼠角膜 IL-6的表达水平。结果：对未进行碱烧伤的角膜使用托珠单抗治疗未观察到明显的角膜结构的损伤。角膜碱烧伤后可见角膜结构的破坏，角膜瘢痕形成并伴有角膜上皮伤口的延迟愈合。使用托珠单抗治疗后，虹膜前粘连的发生率从86.67%下降至20%(P <0.01)，角膜后弹力层脱离的发生率从93.33%下降至53.33%(P <0.05)，角膜厚度小于对照组[(100.03±15.73)μ m vs. (207.02±56.30)μ m，P＜0.001]，角膜混浊评分从对照组的3.76±0.44下降到治疗组的1.94±0.83(P <0.001)，治疗组在第5天(P <0.05)、第10天(P <0.001)和第14天(P <0.001)的上皮愈合率高于对照组。角膜碱烧伤后可见IL-6大量分布于角膜全层，且可见大量肌成纤维细胞形成及免疫细胞浸润，托珠单抗治疗后抑制了IL-6的表达(下降77.5%，P <0.05)，肌成纤维细胞数量从每视野(91.44±65.60)个减少至(12.89±10.51)个(P <0.01)，免疫细胞的数量从每视野(60.30±28.71)个细胞减少至每视野(6.80±3.82)个细胞(P <0.001)。此外，托珠单抗还减少角膜切片中每视野的杯状细胞数目由(11.3±5.29)个减少至(2.0±1.90个)(P <0.01)，并减少角膜新生血管和新生淋巴管的形成(分别减少了76.86%和71.16%，均P <0.001)。结论：局部使用托珠单抗抑制IL-6未见明显角膜毒性，且可以调控角膜碱烧伤后的修复。

Objective: To evaluate the safety and effect of topical IL-6 inhibitor tocilizumab eye drops in regulating corneal alkali burn repair. Methods: Six mice without corneal burns were locally treated with tocilizumab eye drops (2.5 mg/mL) and six mice with corneal pseudo burn were treated with saline, respectively, as experimental and blank groups to evaluate the safety of tocilizumab eye drops. 30 alkali burned mice were randomly divided into a treatment group and a control group in a 1:1 ratio. The treatment group received tocilizumab eye drops, while the control group received physiological saline solution 6 times per day for 14 days. Observe the anterior adhesion of the iris, detachment of the Descemet membrane, and corneal edema through anterior segment optical coherence tomography (AS-OCT), and examine corneal scarring and epithelial wound healing under a stereomicroscope. Evaluate IL-6 localization, myofibroblasts, immune cell infiltration, and corneal epithelial metaplasia on corneal sections. Evaluate corneal neovascularization and neovascularization area by whole-mount cornea staining. Detect the expression level of IL-6 in mouse cornea by qRT-PCR. Results: No significant damage to the corneal structure was observed in the treatment of unburned corneas with tocilizumab. After corneal alkali burns, the corneal structure was damaged, corneal scarring was formed, and delayed healing of corneal epithelial wounds was observed.After treatment with tocilizumab, the incidence of anterior synechia of the iris significantly decreased from 86.67% to 20% (P <0.01), the incidence of Descemet membrane detachment decreased from 93.33% to 53.33% (P <0.05), the corneal thickness was significantly less than that of the control group (100.03±15.73) μ m vs. (207.02±56.30)μ m (P <0.001), the corneal opacity score decreased from 3.76±0.44 in the control group to 1.94±0.83 in the treatment group (P <0.001), and the epithelial healing rate in the treatment group was significantly higher than that in the control group on day 5 (P <0.05), day 10 (P <0.001), and day 14 (P <0.001).After corneal alkali burns, IL-6 was distributed throughout the corneal layer, and a large number of myofibroblasts and immune cells were observed. After treatment with tocilizumab, the expression of IL-6 was inhibited (decreased by 77.5%, P <0.05), the number of myofibroblasts decreased from (91.44±65.60) per field to (12.89±10.51) per field (P <0.01), and the number of immune cells decreased from (60.30±28.71) cells per field to (6.80±3.82) cells per field (P <0.001). In addition, tocilizumab also reduced the number of goblet cells per field in corneal sections (from 11.3±5.29 to 2.0±1.90) (P <0.01), and reduced the formation of corneal neovascularization and neovascular lymphatic vessels (by 76.86% and 71.16%, respectively, both P <0.001). Conclusion: Topical use of tocilizumab to inhibit IL-6 showed no significant corneal toxicity and can regulate the repair of cornea after alkali burns.

目的:探讨碱性成纤维细胞生长因子(Basic fibroblast growthfacfor, bFCF)，表皮细胞生长因子(Epidermal growth factor, EGF)和神经细胞生长因子(Nerve growth factor, NGF)对体外培养的人角膜内皮细胞的生长调控作用。

方法:将相同数量的人角膜内皮细胞接种于96孔板。加人浓度分别为0 ng/ml、1 ng/ml、3 ng/ml、10 ng/ml、30 ng/ml、100 ng/ml的 EGF、bFGF 和 NGF 进行培养。5 天后 MTT法用检测增殖情况。
结果:在0 ng/ml、1 ng/ml、3 ng/ml、10 ng/ml、30 ng/ml、100 ng/ml 浓度下 bFGF 组的平均 OD 值分别为： 0.224±0.045、0.239±0.040、0.262±0.0342、0.278±0.0319、0.281±0.0324、0.260±0.0310 EGF组的平均 0D 值分别为: 0.228±0.0304、0.245±0.0418、0.267±0.0454、0.275±0.0347、0.271±0.0449、0.250±0.0253。NGF 组的平均 OD 值分别为：0.216±0.0187、0.228±0.0226、0.231±0.0225、0.242±0.0279、0.245±0.0294、0.247±0.0349。
结论:bFGF在 30 ng/ml范围内对内皮细胞生长有促进作用,并具有剂量依赖性。高于100 ng/ml时促生长作用降低。EGF在10 ng/ml范围内对内皮细胞生长有促进作用，并具有剂量依赖性。高于30 ng/ml 时促生长作用降低。NGF本次实验剂量范围内对角膜内皮细胞生长无明显作用。

Purpose: To investigate effect of bFGF, EGF and NGF on growth of cultured humancorneal endothelial cells.
Methods: Cultured human corneal endothelial cells were seeded into individual wellsof 96-well tissue culture plate with the same culture media containing separately bFCF, EGF or NGF with a serial of concentrations of 0 ng/ml、1 ng/ml 、3 ng/ml、10 ng/ml 、30 ng/ml and 100 ng/ml and then cultured for 5 days. Then MTT method wasused to detect the growth of the cells.
Results: The averaged OD values of the cell wells containing bFCF with a serial of concentrations of 0 ng/ml、1 ng/ml、3 ng/ml、10 ng/ml、30 ng/ml and 100 ng/ml were 0.224±0.045, 0.239±0.040, 0.262±0.0342, 0.278±0.0319, 0.281±0.0324, 0.260±0.0310. The averaged OD values of EGF group and NGF group were separately 0.228±0.0304，0.245±0.0418, 0.267±0.0454, 0.275±0.0347, 0.271±0.0449, 0.250±0.0253 and 0.216±0.0187, 0.228±0.0226, 0.231±0.0225, 0.242±0.0279, 0.245±0.0294，0.247±0.0349.
Conclusion: bGFC can promote the growth of human corneal endothelial cells in adose dependent manner while with concentration lower than 30 ng/ml. bFGF withconcentration that is higher than 100ng/ml will weaken this effect. EGF can alsopromote proliferation of human corneal endothelial cells demonstrating a linear dosedependent effect when its concentration is lower than 10 ng/ml and this effect decreasedwhen its concentration was higher than 30 ng/ml, NGF showed no effect on the growthof human cornea endothelial cells in this study.