目的：探讨越橘提取物对视网膜色素上皮(retinal pigment epithelial,RPE)细胞氧化损伤和血管生成的抑制作用。方法：① 将人RPE细胞(ARPE-19)分为对照组、模型组、越橘提取物组，模型组给予0.5 mmol/L3-吗啉代亚胺 (3 morpholinosydnonimine,SIN-1)处理24 h，越橘提取物组给予10ng/mL越橘提取物处理1h 后，再给予0.5 mmol/L SIN-1处理24 h。细胞活性检测法(Cell Counting Kit-8,CCK-8)测定各组细胞活性，流式细胞术检测细胞活性氧(reactive oxygen species,ROS)水平。②将人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)分为对照组、模型组、越橘提取物组，模型组给予10 ng/mLVEGF处理8 h，越橘提取物组给予10 ng/mL越橘提取物处理12 h后，再给予10 ng/mLVEGF处理8 h。划痕实验和Transwell实验检测细胞迁移和侵袭，管腔形成实验检测细胞血管生成情况。结果：①越橘提取物在质量浓度≤10 ng/mL对 ARPE-19细胞无明显毒性。给予SIN-1处理后，ARPE-19细胞活性明显降低，而预孵10 ng/mL越橘提取物可使细胞活性恢复接近至正常(P<0.001)，后续实验均选用该浓度。同时，越橘提取物可降低SIN-1诱导的ARPE-19细胞内ROS水平(P<0.0001)。②给予VEGF处理后，HUVEC细胞迁移和侵袭能力增强，而预孵越橘提取物可使细胞迁移和侵袭能力减弱(P<0.0010)。同时，越橘提取物可抑制VEGF诱导的HUVEC细胞小管形成(P<0.0001)。结论：越橘提取物具有较强的抗氧化能力和抗血管生成活性，为其作为潜在治疗年龄相关性黄斑变性治疗药物提供了科学依据。
Objective: To investigate the inhibitory effects of bilberry extract on oxidative damage and angiogenesis in retinal pigment epithelial (RPE) cells. Methods: Human RPE cells (ARPE-19) were divided into control, model and bilberry extract groups. The model group was treated with 0.5 mmol/L SIN-1 for 24 h, and the bilberry extract group was treated with 10ng/bilberry extract for 1 hour, followed by 0.5mmol/L SIN-1 for 24 h. Cell viability was measured by CCK-8. Level of reactive oxygen species (ROS) was determined using flow cytometry. Human umbilical vein endothelial cells (HUVEC) were divided into three groups, control group, model group and bilberry extract group. The model group was treated with 10ng/mL VEGF for 8 h, and the bilberry extract group was treated with 10 ng/mL bilberry extract for 12 h, followed by 10ng/mLVEGF for 8 h. Cell migration and invasion were measured by wound healing assay and Transwell assay. Cell angiogenesis was determined by tube formation assay. Results: 1. Bilberry extract had no obvious toxicity to ARPE-19 cells(≤10 ng/mL). AfterSIN-1 treatment , the the viability of ARPE-19 cells was significantly reduced, while incubation with 10 ng/ml bilberry extract could restore cell activity to the normal levels(P<0.001). This concentration was selected for subsequent experiments. Additionally, bilberry extract reduced the level of ROS in SIN-1-induced ARPE-19 cells(P<0.0001). VEGF treatment significantly enhanced the migration and invasion of HUVEC, whil epretreatment with bilberry extract attenuated these effects(P<0.0010). Meanwhile, bilberry extract could significantly inhibit VEGF-induced tube formation in HUVEC(P<0.0001). Conclusion: Bilbery extract possess strongantioxidant and anti-angiogenetic activities, suggesting its potential as treatment agent for AMD.
甲氨蝶呤(Methotrexate,MTX)是一种叶酸类似物，具有抗增殖、抗炎和免疫调节作用。在临床上广泛用于治疗多种疾病，包括恶性肿瘤和自身免疫性疾病。MTX通过眼局部注射或者全身给药用于眼科疾病的治疗，其安全性和有效性均已被证实。其中玻璃体腔注射可以减少糖皮质激素及免疫抑制剂的全身使用，在眼科的应用越来越广泛。MTX在备孕期男性及女性、妊娠期妇女、哺乳期妇女、儿童、青少年及老年人等特殊患者人群中的应用有其特殊性，用药情况因患者而异。低剂量MTX常见不良反应包括胃肠道症状、肝功能异常等，比较少出现严重的不良反应，但仍应做好药学监护。该篇综述总结MTX的临床使用方法及其不良反应，为其在眼科的临床应用提供参考。
Methotrexate (MTX) is a folate analog with anti-proliferative, anti-inflammatory and immunomodulatory effects.It is widely used in clinical practice to treat various diseases, including malignant tumors and autoimmune diseases.MTX is used for treating eye diseases by local injection or whole-body administration. Its safety and efficacy have been confirmed. The intravitreal injection can reduce the whole-body use of glucocorticoids and immunosuppressive agents, so it is applied in ophthalmology increasingly. The specificity of MTX has been found in its application in special patients, such as men and women in pregnancy preparation period, pregnant women, lactating women, children, adolescents and the elderly. The dosage varies from patient to patient. Common adverse reactions to low-dose MTX include gastrointestinal symptoms, liver dysfunction, etc. Serious adverse reactions are relatively rare, but pharmaceutical monitoring is still necessarily needed. This review summaries the usage and adverse reactions of MTX in clinical practice, to provide reference for its application in clinical practice in ophthalmology.
术前有效杀灭或减少眼表微生物的数量是降低术后眼内炎的重要措施。聚维酮碘消毒液和妥布霉素均能有效的降低眼科手术术后眼内炎发生率。笔者通过文献报道，综合分析聚维酮碘消毒液及妥布霉素滴眼液及注射液对眼科手术结膜囊微生物的控制能力，为眼科手术术前准备提供理论依据。
Effectively killing or reducing the number of ocular microbes before surgery is an important measure to reduce postoperative endophthalmitis. Both povidone-iodine disinfectant and tobramycin can effectively reduce the incidence of endophthalmitis after ophthalmic surgery. Through literature reports, the author comprehensively analyzes the control ability of povidone-iodine disinfectant and tobramycin eye drops and injection on the microscopy of conjunctival sac in ophthalmic surgery, and provides theoretical basis for preoperative preparation of ophthalmic surgery
目的：建立高效液相色谱法(high-performance liquid chromatography，HPLC)测定盐酸丁卡因滴眼液中盐酸丁卡因和羟苯乙酯的含量。方法：采用Agilent Eclipse PLUS C18色谱柱(250 mm ×4.6 mm，5 μm)；以1%三乙胺溶液(三乙胺10 mL，加水990 mL，用冰醋酸调节pH值至5.0±0.5)-乙腈(65 : 35)为流动相，流速为1.0 mL/min；柱温为30 ℃；检测波长为256 nm；进样体积20 μL。结果：盐酸丁卡因在0.05~0.36 mg/mL范围内线性关系良好( r =1.000)，平均回收率为99.2%，相对标准偏差(relative standard deviation，RSD)为0.3%(n=9)，羟苯乙酯在3.02~24.14 μg/mL范围内线性关系良好(r=1.000)，平均回收率为98.2%，RSD为0.4%(n=9)，该方法重现性好。对照品溶液和供试品溶液在室温放置24 h基本稳定；结论：本方法简便、快速、准确。适用于检测盐酸丁卡因滴眼液中盐酸丁卡因和羟苯乙酯的含量。
Objective: To establish a high-performance liquid chromatography (HPLC) method for the determination of tetracaine hydrochloride and ethyl hydrobenzoate in tetracaine hydrochloride eye drops. Methods: The analytic column was Agilent Eclipse Plus C18 (4.6 mm ×250 mm, 5 μm) and the mobile phase was 1% triethylamine solution (10 mL triethylamine and 990 mL water, pH adjusted to 5.0±0.5 with glacial acetic acid) - acetonitrile (65:35) at a flow rate of 1.0 mL/min. The detection wavelength was 256 nm and the column temperature was 30 ℃. The injection volume was 20 μL. Results: The linear range of tetracaine hydrochloride was well shown within 0.05–0.36 mg/mL (r=1.000). The average recovery rate of tetracaine hydrochloride was 99.2% with relative standard deviation (RSD) 0.3% (n=9). The linear range of ethyl hydrobenzoate was well shown within 3.02–24.14 μg/mL (r=1.000). The average recovery rate of tetracaine hydrochloride was 98.2% with RSD 0.4%(n=9). The method had high reproducibility. The reference solution and testing solution were stable for 24 h in room. Conclusion: The method is simple, quick and accurate, which is suitable for the determination of tetracaine hydrochloride and ethyl hydrobenzoate in tetracaine hydrochloride eye drops.
