短波视蛋白在豚鼠频闪光诱导性近视和形觉剥夺近视眼模型中的表达差异

目的观察豚鼠频闪光诱导性近视和形觉剥夺近视模型中短波视蛋白(S-opsin)表达差异,并初步探讨原因。方法 36只普通级2周龄豚鼠随机分成三组:频闪组(FLM组,n=13),形觉剥夺组(FDM组,n=12),对照组(n=11)。FLM组,饲养笼具安装有频闪仪(频率0.5 Hz),笼具内装有发光二极管;FDM组豚鼠右眼用半透明眼罩遮盖,并确保豚鼠眼睑能正常活动;对照组豚鼠不予特殊处理。在造模第1天(0周)和第6周测量豚鼠右眼屈光度、眼轴长度和角膜曲率半径,并通过免疫荧光法观察S-opsin表达。结果第0周,FLM、FDM组与对照组屈光度、眼轴长度、角膜曲率半径差异均无显著性(P0.05)。造模6周后,与对照组相比,FLM组、FDM组屈光度变化值、眼轴长度变化值差异均有显著性(P0.05),而角膜曲率半径变化值差异无显著性(P=0.358),提示成功建立近视模型。FLM组与FDM组相比,屈光度变化值、眼轴长度变化值、角膜曲率半径变化值差异均无显著性(P0.05)。免疫荧光结果显示:FLM组视蛋白灰度值对照组视蛋白灰度值FDM组视蛋白灰度值,任意两组进行比较,差异均有显著性(P0.001)。结论频闪光和形觉剥夺均能建立近视模型,频闪光诱导性近视模型中S-opsin产生增加,而形觉剥夺性近视模型中S-opsin产生减少,说明两种近视模型的发生机制可能不同。     

孕14-19+6周正常胎儿胼周动脉的超声评估

摘要 目的：探讨应用超声SlowflowHD（超低速血流成像）联合RadiantFlow（二维立体血流成像）技术评估孕14-19⁺⁶周正常胎儿胼周动脉的可行性，观察胎儿胼周动脉及分支走行，测量其长度和高度，并探讨二者与孕周及双顶径的关系。方法：纳入2022年5月-2023年3月来南京医科大学附属苏州医院进行常规超声检查的150例孕14-19⁺⁶周孕妇，在胎儿正中矢状切面上获取胎儿胼周动脉及分支的图像，测量其长度和高度，测量重复3次，取平均值，并观察胎儿胼周动脉及分支走行。所有孕妇均随访至中孕晚期（20-28周）或晚孕期行产前超声检查未见胼胝体异常及其他颅内结构异常。采用pearson相关分析和回归分析判断胎儿胼周动脉长度及高度与孕周及双顶径的关系。结果：胎儿胼周动脉长度及高度随着孕周及双顶径的增加而增加（P＜0.05）。胎儿胼周动脉长度与孕周、双顶径之间呈线性正相关，且胎儿胼周动高度与孕周、双顶径之间呈线性正相关（P＜0.05）。在孕14-19⁺⁶周中所有111病例均类"C"形走行，额前内侧动脉、额中内侧动脉、额后内侧动脉三者几乎与胎儿胼周动脉垂直，类"鸡冠"，三者显示率100%；旁中央动脉在孕14-15⁺⁶周、16-17⁺⁶周、18-19⁺⁶周显示率分别约53.1%（17/32）、88.2%（45/51）、92.8%（26/28）；楔前动脉在孕14-15⁺⁶周、16-17⁺⁶周、18-19⁺⁶周显示率分别约3.1%（1/32）、35.2%（18/51）、78.5%（22/28）。结论：应用超声SlowflowHD（超低速血流成像）联合RadiantFlow（二维立体血流成像）技术显示孕14-19⁺⁶周正常胎儿胼周动脉及分支具有可行性，胎儿胼周动脉长度和高度与孕周及双顶径呈一定的线性相关。     

天津市6～18岁青少年近视患病率调查及其影响因素Logistic回归分析

摘要 目的：调查分析天津市6～18岁青少年近视患病率及其影响因素。方法：采用整群抽样方法抽取天津市6～18岁在校学生2230例作为研究对象。对所有研究对象均进行视力以及屈光度等指标的检测,同时通过调查问卷采集其基本资料。将所有研究对象按照是否近视分成近视组以及非近视组,以多因素Logistic回归分析明确天津市6～18岁青少年近视的影响因素。此外,比较两组青少年的血清微量元素水平。结果：2230例青少年近视患病率为41.12%,且6～9岁、10～12岁、13～15岁、16～18岁青少年的近视患病率呈逐渐升高趋势,组间对比差异有统计学意义（均P＜0.05）。近视组年龄、近距离用眼时间、使用电子设备时间以及父母一方或双方近视人数占比均高于非近视组,而户外活动时间短于非近视组（均P＜0.05）。多因素Logistic回归分析可得：天津市6～18岁青少年近视的危险因素包括年龄、父母一方或双方近视、近距离用眼时间、使用电子设备时间,保护因素为户外活动时间（均P＜0.05）。近视组血清Zn²⁺、Fe²⁺水平低于非近视组,而Cu²⁺水平高于非近视组（均P＜0.05）。结论：天津市6～18岁青少年近视患病率较高,其影响因素包括年龄、父母一方或双方近视、近距离用眼时间、使用电子设备时间以及户外活动时间,且近视青少年存在微量元素缺乏的情况,值得临床重点关注。     

530nm单色光诱导豚鼠近视眼模型的建立

目的应用530 nm单色光光照建立一种新型近视眼动物模型。方法20只约2周龄健康雄性豚鼠,随机分成两组（n=10）,实验组和对照组分别在绿光（530 nm）和白光（色温5000 k）下进行饲养。设置照明参数：光量子数相同,为每秒3×10-4μmol/cm2;实测光强度绿光为0.150 mW/cm2,白光为0.247 mW/cm2。实验前每组进行眼球生物学测量（屈光度、角膜曲率、眼轴各部分长度）,光照后12周重复测量以上数据,每只豚鼠均取右侧眼参数进行统计分析。结果光照前两组生物学测量参数差异无显著性。光照12周后,绿光组屈光度发生-3.125±0.76 D的变化,白光组为-1.075±0.71D,绿光组同对照白光组相比平均形成约2.0 D的近视,差异有显著性;绿光组眼轴和玻璃体腔分别增长0.98±0.13 mm与0.33±0.14 mm,对照组分别为0.77±0.22 mm与0.13±0.14 mm,绿光组较对照组眼轴和玻璃体腔长度延长较快,差异有显著性;光照后两组角膜曲率半径、前房深度和晶状体厚度均发生不同程度增加,但两组间变化差异无显著性。结论530 nm单色光诱导豚鼠眼球眼轴和玻璃体腔长度延长较快,产生近视.     

苦绳（Dregea sinensis）中多氧化孕烷糖苷与免疫蛋白相互作用的研究

目的 基于化学结构特点研究植物苦绳（Dregea sinensis）中多氧孕烷糖苷的生物活性,以及化合物分子与靶蛋白分子间的作用机理。方法 本研究采用分子对接和表面等离子体共振方法,对191个多氧化孕烷糖苷类成分（>800 u）进行了免疫活性及其与免疫蛋白的动力学评价。结果 通过分子对接方法筛选出7个配体分子（6、18、23、30、78、79和80）和3个免疫相关蛋白（IL-2Rα、TLR4和TNF-α）。研究表明,化合物30和78在SPR实验中与靶蛋白IL-2Rα和TLR4具有显著的结合趋势,与IL-2Rα的结合常数K_D分别为2.41×10^-6和2.14×10^-6 mol/L,与TLR4则分别为1.96×10^-5和5.60×10^-6 mol/L。分子动力学研究进一步表征SPR阳性分子6、18、30、78和80与靶蛋白之间的相互作用基团。结论 研究揭示多氧化孕烷糖苷类成分可以通过形成氢键和Pi-Pi相互作用与靶蛋白结合。研究内容对快速评价多氧化孕烷糖苷类成分的活性具有重要意义,为揭示低丰度药效物质的潜在作用机制进行了有益探索。     

依托咪酯靶控输注联合右美托咪定对胃癌根治术患者应激反应、炎症因子和细胞免疫功能的影响

摘要 目的：观察胃癌根治术患者在依托咪酯靶控输注联合右美托咪定麻醉下,机体细胞免疫功能、炎症因子、应激反应的影响。方法：选取2021年4月~2022年6月期间我院收治的择期行胃癌根治术患者100例,按照信封抽签的形式将患者分为对照组（n=50）和观察组（n=50）,对照组患者麻醉选用依托咪酯注射液靶控输注,观察组则在对照组的基础上结合静脉输注盐酸右美托咪定注射液。观察两组患者不良反应发生情况、血流动力学指标[心率（HR）、收缩压（SBP）、舒张压（DBP）]、应激反应指标[皮质醇（Cor）、肾上腺素（E）、促肾上腺皮质激素（ACTH）]、炎症因子[肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）、超敏C反应蛋白（hs-CRP）]、细胞免疫功能指标。结果：观察组用药时、拔管时SBP、HR、DBP低于对照组（P<0.05）。两组不良反应发生率组间对比无差异（P>0.05）。观察组术后1 d E、IL-6、Cor、TNF-α、ACTH、 hs-CRP、 CD8⁺低于对照组（P<0.05）。观察组术后1 d CD4⁺、CD3⁺、CD4⁺/CD8⁺高于对照组（P<0.05）。结论：胃癌根治术患者使用依托咪酯靶控输注联合右美托咪定麻醉,血流动力学、应激反应可得到有效控制,炎症因子、免疫功能的影响也可大大减轻,利于手术的顺利进行。     

消疬膏对脾胃虚弱证颈部淋巴结核患者CD4+CD25highFoxP3+调节性T淋巴细胞和胃肠激素的影响

摘要 目的：研究消疬膏对脾胃虚弱证颈部淋巴结核患者CD4⁺CD25^highFoxP3⁺调节性T淋巴细胞和胃肠激素的影响。方法：选择南京市中西医结合医院于2020年3月~2022年1月期间收治的颈部淋巴结核患者120例。按照随机数字表法分为对照组（60例）和研究组（60例）。对照组接受常规抗结核方案，研究组接受消疬膏联合常规抗结核方案，两组疗程均为6个月。观察两组治疗前和治疗6个月后的中医证候积分、CD4⁺CD25^highFoxP3⁺调节性T淋巴细胞相关指标和胃肠激素的变化，记录两组治疗期间用药安全性。结果：两组治疗6个月后肿疡大小、肿疡皮色、面色、纳呆、倦怠乏力、便溏、舌质、脉象评分均下降，且研究组低于对照组（P<0.05）。研究组治疗6个月后CD4⁺CD25^highFoxP3⁺调节性T淋巴细胞、白介素-10（IL-10）水平低于对照组，γ-干扰素（IFN-γ）水平高于对照组（P<0.05）。研究组治疗6个月后胆囊收缩素（CCK）、胃泌素（GAS）水平低于对照组，胃动素（MTL）水平高于对照组（P<0.05）。研究组的不良反应总发生率低于对照组（P<0.05）。结论：消疬膏治疗脾胃虚弱证颈部淋巴结核患者，可有效改善患者临床症状，可能与调节CD4⁺CD25^highFoxP3⁺调节性T淋巴细胞和胃肠激素水平有关。     

Chk2在Bmi1缺失所致的肾脏早衰和纤维化中的作用和机制研究

摘要 目的：探究细胞周期检测点激酶2（cell-cycle checkpoint kinase 2，Chk2）在B淋巴瘤Mo-MLV插入区1（B cell-specific MLV integration site-1，Bmi1）缺失所致的肾脏早衰和纤维化中的作用及可能的机制。方法：取5周龄WT、Bmi1^-/-、Chk2^-/-、Bmi1^-/-Chk2^-/-小鼠肾脏，采用HE染色观察肾脏结构变化，采用免疫荧光和Masson染色观察肾脏纤维化情况，采用衰老相关β半乳糖苷酶（Senescence-associated β-galactosidase，SA-β-gal）染色观察肾脏衰老情况，采用免疫组化染色和western blot观察肾脏超氧化物歧化酶1（Superoxide Dismutase 1，SOD1）、超氧化物歧化酶2（Superoxide Dismutase 2，SOD2）表达水平和定位。从5周龄WT、Bmi1^-/-、Chk2^-/-、Bmi1^-/-Chk2^-/-小鼠肾脏皮质中提取和分离原代肾小管上皮细胞，采用免疫荧光和western blot检测其SOD1、SOD2表达水平。结果：与WT小鼠肾脏组织相比，Bmi1^-/-小鼠肾脏组织表现为体积变小、肾脏皮质厚度减少、肾小球数量减少，β-gal活性增加，SOD1和SOD2水平降低；与Bmi1^-/-小鼠肾脏相比，Bmi1^-/-Chk2^-/-小鼠肾脏体积增大、肾脏皮质厚度增加，肾小球数量增多，β-gal活性降低，SOD1和SOD2水平增加。与WT小鼠肾脏皮质原代肾小管上皮细胞相比，Bmi1^-/-小鼠肾脏皮质原代肾小管上皮细胞中SOD1和SOD2水平；与Bmi1^-/-小鼠肾脏皮质原代肾小管上皮细胞相比，Bmi1^-/-Chk2^-/-小鼠肾脏皮质原代肾小管上皮细胞中抗氧化指标SOD1和SOD2增加。结论：Chk2通过抑制肾小管上皮细胞的抗氧化能力促进Bmi1缺失所致的肾脏早衰和纤维化。     

广州地区育龄人群地中海贫血基因分型及筛查漏检误诊的原因分析

摘要 目的：了解广州地区育龄人群的地中海贫血（简称：地贫）基因携带率及基因型分布特征,分析地贫筛查的漏检和误诊原因,为育龄地贫基因携带者进行人工辅助受孕提供依据。方法：收集2019年1月到2021年12月在我院生殖医学科就诊并同时进行地贫筛查及地贫基因检测的育龄患者31455例,分析其地贫相关实验室检查结果,包括红细胞参数、红细胞渗透脆性、血红蛋白电泳及地贫基因检测,计算地贫基因携带率,并通过比较筛查和基因检测结果找出漏筛和误诊病例。结果：共检出育龄地贫基因携带者4455例,地贫基因携带率为14.16 %。其中,α-地中海贫血3365例,常见的基因型有-- ^SEA /αα、-α^3.7/αα和 -α^4.2/αα;β-地中海贫血914例,常见的基因型有β^{CD41-42(-TCCT)} /β ^N 、β^{IVS-2-654(C→T)} /β ^N 和β^-28(A→G) /β ^N ;α-合并β-地中海贫血176例,最常见的基因型是β^{CD41-42(-TCCT)} /β ^N /-- ^SEA /αα。漏筛病例有731例,多为静止型α-地贫。误诊病例有4784例,其中有2701例误诊病例其红细胞渗透脆性和红细胞参数均正常,仅因HbA2<2.5 %且HbA>97.5 %被误诊为携带地贫基因。结论：广州地区的育龄人群具有较高的地贫基因携带率,静止型α-地贫容易在筛查中被漏检。     

组蛋白去乙酰化酶3对外周CD4+ T细胞分化的影响

目的 探讨组蛋白去乙酰化酶3（HDAC3）对外周CD4⁺ T细胞分化及功能的调控作用。方法 采用CD4cre酶介导Hdac3杂合基因缺失小鼠（Hdac3^fl/flCD4^cre+/-）及其野生型正常对照小鼠（Hdac3^fl/fl，WT），流式细胞术检测HDAC3缺失对外周CD4⁺和CD8⁺ T细胞比例和数量的影响；在体外佛波酯（PMA）和离子霉素（Ionomycin）刺激条件下，流式细胞术检测HDAC3缺失对CD4⁺ T细胞中IFN-γ、IL-4和IL-17A的表达以及Tfh细胞产生的影响；采用ELISA检测HDAC3缺失对小鼠血清IFN-γ、IL-4和IL-17表达的影响；分选Hdac3^fl/flCD4^cre+/-和WT小鼠外周初始CD4⁺ T细胞，分别在Th1和Th2分化条件下培养，细胞内染色检测HDAC3缺失对Th1、Th2以及Th17相关细胞因子及其特异转录因子表达的影响；采用Microarray检测HDAC3缺失对CD4⁺ T细胞分化亚群相关基因表达的影响；采用链脲佐菌素（STZ）处理小鼠构建I型糖尿病（TIDM）疾病模型，检测HDAC3缺失对T1DM发病的影响。结果 与WT小鼠相比，Hdac3^fl/flCD4^cre+/-小鼠外周CD4⁺和CD8⁺ T细胞的比例和数量显著降低。Hdac3^fl/flCD4^cre+/-小鼠CD4⁺ T细胞及血清中IFN-γ的表达显著降低，而IL-4和IL-17A的表达显著增加，Tfh细胞比例也显著增加；HDAC3缺失抑制体外培养CD4⁺ T细胞向Th1分化但促进其向Th2分化；Microarray检测发现HDAC3缺失导致Th1型细胞谱系基因表达降低，而Th2、Th17以及Tfh细胞谱系基因表达增加；在STZ诱导条件下，HDAC3缺失抑制小鼠T1DM的发生和CD4⁺ T细胞向Th1分化。结论 HDAC3促进外周CD4⁺ T细胞向Th1细胞分化并加重T1DM的发生。     

The measurement of ocular axial length in normal human eyes based on an improved Twyman-Green interferometer

In order to meet the demands of myopia prevention, as well as the increasing needs of measurement for refractive and cataract operations in China, a new axial length (AL) measurement system combining an improved Twyman-Green interferometer with digital signal processing has been established. The ALs of 33 eyes of different optical refractive subjects (−8.50 ~ 0.50 D) were measured with the New AL and intraocular lens (IOL) master. The repeatability of measurements with the New AL was assessed using coefficient of variation (CoV) and intra-group correlation coefficient (ICC). Comparison, correlation, linear regression and agreement of AL between devices were analyzed. There was good repeatability (CoV = 0.0617%, ICC = 0.9999) with the New AL and great agreement has been obtained with both devices. These show that the New AL is capable of providing precise AL values over the normal AL range compared to the IOL master, and indicate that the New AL developed can be used for routine clinical AL measurements.     

Sources of variation affecting cashmere grown in the Pamir mountain districts of Tajikistan and implications for industry development

We aimed to quantify the sources of variation contributing to the main quality attributes of cashmere produced from goats in the Pamir mountain districts of Murghab, Shugnon and Vanj in Tajikistan. In early spring 2010, mid-side samples were taken from 194 adult females, 43 adult males and 20 castrates belonging to 58 farmers and pastoralists in 14 villages. For 57 goats, samples were also taken from the shoulder and hip sites. Mean fibre diameter (MFD), fibre curvature (FC) and cashmere staple length (SL) data were examined using a general linear model to determine the relationships between fleece attributes and other possible effects. For females, the mean (s.d.) for MFD, FC and SL were: 16.5 (1.70) μm; 46 (12.1)°/mm; 53 (22.9) mm. MFD was affected by district, SL and age of goat. SL was affected by district, MFD, gender, age of goat and village. FC was affected by district, MFD, shade of cashmere, age of goat and farmer. Cashmere from Vanj district was finer and shorter than cashmere from Murghab and Shugnon. Cashmere grown on the mid-side and hip sites was finer and had higher FC than cashmere grown on the shoulder. Cashmere grown on the hip was shorter than cashmere grown on the mid-side and shoulder. About 50% of the cashmere sampled was < 16.4 μm and potentially suitable for knitwear. Of this fine cashmere, 53% was 34 mm or longer. A further 37% of the cashmere was 16.4-18.5 μm, and suitable for weaving as 97% was longer than 36 mm. Almost 12% of samples were > 18.5 μm and may only be suitable for weaving or, if cashgora, will have little commercial value. Most of the cashmere was coloured. There are cashmere goats in the Murghab, Shugnon and Vanj districts of Tajikistan which produce the finest qualities of cashmere, comparable to premium grades of Chinese cashmere. There is substantial scope to increase the commercial value of cashmere produced by goats in Tajikistan, in particular increasing SL for fine cashmere, reducing MFD for the longest cashmere and ensuring cashmere has acceptable FC and white colour.     

Swelling of the collagen-keratocyte matrix of the bovine corneal stroma ex vivo in various solutions and its relationship to tissue thickness

AIM: The mammalian corneal stroma, like some other connective tissues, can absorb fluid, swell and become oedematous. Since studies on the corneal stroma have been carried out with different types of preparations and solutions, inter-study comparisons are very difficult. A study was thus undertaken on a standardised preparation to assess the relative magnitude of this swelling and its relationship to thickness of the preparations. METHODS: From selected recent post-mortem eyes of adult cattle, stroma preparations were cut from the central part of the cornea. These preparations were immersed in various solutions of known pH and osmolality, and the time-dependent changes in wet mass were assessed over 9 h at 37 degrees C. The relative rates and magnitude of the swelling of the tissue were then compared. RESULTS: A reference value for stromal swelling was obtained by incubation in a 35 mM bicarbonate-buffered mixed salts solution equilibrated with 5% CO2-air (pH 7.60) where a 3.39-fold increase in wet mass and a 4.58-fold increase in thickness was realised in 9 h, at an initial rate of 76 +/- 3%/h. The swelling was essentially the same in an organic buffer-mixed salt solution (pH 7.5) but progressively greater in phosphate-buffered saline (pH 7.5), a range of phosphate buffers (10-67 mM, pH 7.5), NaCl solutions (0.025-1%) and with gross swelling observed in water (where a 15.9-fold increase in wet mass occurred along with a 25-fold increase in thickness, at an initial rate of 643 +/- 62%/h). Overall, the wet mass changes were strongly related to thickness (P < 0.001). CONCLUSIONS: The results confirm that the selection of solution(s) for studies on corneal stromal swelling is critical. The swelling (oedema) is lower in a physiologically-relevant solution (similar to the aqueous humour of the eye). This indicates that the swelling tendency of the corneal stroma has been overestimated in the past, and that a similar discrepancy may also exist for studies on other connective tissues ex vivo when non-physiological experimental solutions are used.     

Contribution of the intrinsic curvature to measured DNA persistence length

The persistence length of DNA, a, depends both on the intrinsic curvature of the double helix and on the thermal fluctuations of the angles between adjacent base-pairs. We have evaluated two contributions to the value of a by comparing measured values of a for DNA containing a generic sequence and for an "intrinsically straight" DNA. In each 10 bp segment of the intrinsically straight DNA an initial sequence of five bases is repeated in the sequence of the second five bases, so any bends in the first half of the segment are compensated by bends in the opposite direction in the second half. The value of a for the latter DNA depends, to a good approximation, on thermal fluctuations only; there is no intrinsic curvature. The values of a were obtained from measurements of the cyclization efficiency for short DNA fragments, about 200 bp in length. This method determines the persistence length of DNA with exceptional accuracy, due to the very strong dependence of the cyclization efficiency of short fragments on the value of a. We find that the values of a for the two types of DNA fragment are very close and conclude that the contribution of the intrinsic curvature to a is at least 20 times smaller than the contribution of thermal fluctuations. The relationship between this result and the angles between adjacent base-pairs, which specify the intrinsic curvature, is analyzed.     

Environmental monitoring through functional biodiversity tools

The foundational concept for our research, which is largely shared by statisticians and ecologists, is that biodiversity is one of the most important indicators for environmental assessment. Because this indicator decreases in relation to ecosystem stressors, its measurement is essential for predicting future biological impacts of environmental damages. Although many indices have been proposed, no universally accepted measure for biodiversity has yet been established. In this context, the use of diversity profiles allows the analyst to display a family of indices in a single graph. However, this approach presents two critical limitations: first, a community composition is not always interpretable; second, the diversity profiles could lead to ranking issues when the curves intersect each other. The aim of this paper is to resolve these limitations by introducing functional biodiversity tools. In particular, three functional measures are proposed: the derivatives, the radius of curvature and the curve length. The analysis of derivatives and of the radius of curvature addresses the first limitation and highlights the characteristics, the differences and the similarities among communities. Arc length addresses the second limitation, providing a scalar measure that leads to a unique communities ranking for a given pattern of richness even if profiles intersect. The proposed functional models are applied to a real data set involving lichen biodiversity in the province of Genoa, Italy. Our approach allowed us to analyze the characteristics of lichen communities and to identify the biodiversity ranking. The combined use of these tools provides a useful method for identifying areas of high environmental risk, with the potential to address the monitoring of environmental policies.     

光诱导角膜交联及检测技术的研究进展

由于圆锥角膜疾病导致越来越多的人患有近视,常见的矫正方法有佩戴近视眼镜、隐形眼镜等.随着科技的进步,利用光对近视等眼科疾病进行屈光矫正已经成为当前临床中常用的方法.使用光诱导角膜胶原蛋白发生交联,从而达到治疗圆锥角膜疾病、提高患者视力水平的目的,这是一种新型的光治疗眼睛疾病的方法.同时这种方法由于无侵入性、对操作者能力依赖性小等优势成为新的研究热点.本文阐述光诱导角膜交联的基本原理,并介绍其发展历程,分析现有的各种交联方法和角膜检测技术的原理,并对现有交联方法和检测方法的优缺点进行讨论.最后,本文对光诱导角膜交联和检测技术的最新进展进行系统的论述,并对未来的发展趋势进行展望.     

Assessment of the influence of viscoelasticity of cornea in animal ex vivo model using air‐puff optical coherence tomography and corneal hysteresis

Application of the air‐puff swept source optical coherence tomography (SS‐OCT) instrument to determine the influence of viscoelasticity on the relation between overall the air‐puff force and corneal apex displacement of porcine corneas ex vivo is demonstrated. Simultaneous recording of time‐evolution of the tissue displacement and air pulse stimulus allows obtaining valuable information related in part to the mechanical properties of the cornea. A novel approach based on quantitative analysis of the corneal hysteresis of OCT data is presented. The corneal response to the air pulse is assessed for different well‐controlled intraocular pressure (IOP) levels and for the progression of cross‐linking‐induced stiffness of the cornea. Micrometer resolution, fast acquisition and noncontact character of the air‐puff SS‐OCT measurements have potential to improve the in vivo assessment of mechanical properties of the human corneas.      

Absence of corneal endothelium injury in non‐human primates treated with and without ophthalmologic drugs and exposed to 2.8 GHz pulsed microwaves

Microwave‐induced corneal endothelial damage was reported to have a low threshold (2.6 W/kg), and vasoactive ophthalmologic medications lowered the threshold by a factor of 10–0.26 W/kg. In an attempt to confirm these observations, four adult male Rhesus monkeys (Macaca mulatta) under propofol anesthesia were exposed to pulsed microwaves in the far field of a 2.8 GHz signal (1.43 ± 0.06 µs pulse width, 34 Hz pulse repetition frequency, 13.0 mW/cm² spatial and temporal average, and 464 W/cm² spatial and temporal peak (291 W/cm² square wave equivalent) power densities). Corneal‐specific absorption rate was 5.07 W/kg (0.39 W/kg/mW/cm²). The exposure resulted in a 1.0–1.2 °C increase in eyelid temperature. In Experiment I, exposures were 4 h/day, 3 days/week for 3 weeks (nine exposures and 36 h total). In Experiment II, these subjects were pretreated with 0.5% Timolol maleate and 0.005% Xalatan® followed by 3 or 7 4‐h pulsed microwave exposures. Under ketamine–xylazine anesthesia, a non‐contact specular microscope was used to obtain corneal endothelium images, corneal endothelial cell density, and pachymetry at the center and four peripheral areas of the cornea. Ophthalmologic measurements were done before and 7, 30, 90, and 180 days after exposures. Pulsed microwave exposure did not cause alterations in corneal endothelial cell density and corneal thickness with or without ophthalmologic drugs. Therefore, previously reported changes in the cornea exposed to pulsed microwaves were not confirmed at exposure levels that are more than an order of magnitude higher. Bioelectromagnetics 31:324–333, 2010. Published 2010 Wiley‐Liss, Inc.     

Effect of porcine corneal stromal extract on keratocytes from SMILE-derived lenticules

Propagating large amounts of human corneal stromal cells (hCSCs) in vitro while maintaining the physiological quality of their phenotypes is necessary for their application in cell therapy. Here, a novel medium to propagate hCSCs obtained from small incision lenticule extraction (SMILE)-derived lenticules was investigated and the feasibility of intrastromal injection of these hCSCs was assessed. Primary hCSCs were cultured in porcine corneal stroma extract (pCSE) with RIFA medium including ROCK inhibitor Y27632, insulin-transferrin-selenium, fibroblast growth factor 2, L-ascorbate 2-phosphate and 0.5% FBS (RIFA medium + pCSE). Protein profiling of the pCSE was identified using nanoscale liquid chromatography coupled to tandem mass spectrometry (nano LC-MS/MS). After subculturing in RIFA medium + pCSE or 10% FBS normal medium (NM), hCSCs at P4 were transplanted into mouse corneal stroma. Compared with NM, ALDH3A1, keratocan and lumican were significantly more expressed in the RIFA medium + pCSE. ALDH3A1 was also more expressed in the RIFA medium + pCSE than in the RIFA medium. Fibronectin and α-SMA were less expressed in the RIFA medium + pCSE than in the NM. Using Metascape analysis, the pCSE with its anti-fibrosis, pro-proliferation and anti-apoptosis activities, was beneficial for hCSC cultivation. The intrastromally implanted hCSCs in the RIFA medium + pCSE had positive CD34 expression but negative CD45 expression 35 days after injection. We provide a valuable new medium that is advantageous for the proliferation of hCSCs with the properties of physiological keratocytes. Intrastromal injection of hCSCs in RIFA medium + pCSE has the potential for clinical cell therapy.     

Two-year changes in corneal stiffness parameters after accelerated corneal cross-linking

The objective of this non-randomized trial was comparison of two-year changes in dynamic corneal response (DCR) between 18 mW/cm² (5- min) and 9 mW/cm² (10-min) cross-linking (CXL) protocols, using novel stiffness parameters and correlating them to clinical indices. The two groups were evaluated before and 2 years after the procedure using Corvis ST (Oculus Optikgeräte GmbH, Germany) and DCR parameters such as deformation amplitude ratio at 1 mm and 2 mm (DA ratio-1 mm and DA ratio-2 mm) and integrated radius and stiffness parameters at A1 (SP-A1). Two-year follow-up was completed for 16 of the 30 eyes in the 5-min group and 21 of the 25 eyes in the 10-min group; data from those who were lost to follow-up was not included in the analyses. Mean age at baseline was 21.7 ± 4.9 and 21.5 ± 5.2 years in the 5- and 10-min groups, respectively (P = 0.895). At 2 years after CXL, in the 5-min group, the reduction in integrated radius (−1.12 ± 1.27 mm, P = 0.003) was significant, and the increase in SP-A1 (7.11 ± 14.86 mmHg/mm, P = 0.075) was borderline, while in the 10-min group, the decrease in DA ratio-2 mm (−0.43 ± 0.58, P = 0.003) and integrated radius (−1.89 ± 1.72 mm, P < 0.001), and increase in SP-A1 (7.67 ± 10.92 mmHg/mm, P = 0.004) were significant. In both groups, the strongest and significant correlation was observed between DCR parameters and changes in radius of curvature. In conclusion, results indicated corneal strengthening with both protocols especially with the 9 mW/cm². Corvis ST indices can provide “in vivo” biomechanical evidence on the efficacy of CXL that may occur prior to clinical indices.