糖尿病氧化应激环境中α-Klotho对巨噬细胞-血管内皮细胞串扰的影响

目的：探讨糖尿病氧化应激环境中过表达α-Klotho(KL)的小鼠单核巨噬细胞白血病细胞(RAW264.7)对人脐静脉内皮细胞(HUVECs)增生、迁移、管腔形成以及紧密连接的影响。

方法：将RAW264.7细胞分为对照组、4-羟壬二酸酯(4HNE)组、4HNE+KL组,采用免疫荧光实验检测RAW264.7细胞F4/80的表达。制备3组细胞的条件培养基用于培养HUVECs,分为Mø-NC组、Mø-4HNE组和Mø-4HNE+KL组。采用CCK8实验检测血管内皮细胞增生,采用划痕实验和Transwell实验检测迁移,采用管腔形成实验检测管腔形成,采用Western blot实验检测闭合蛋白5(Claudin 5)、咬合蛋白(Occludin)、带状闭合蛋白1(ZO 1)表达水平。

结果：免疫荧光实验结果显示,4HNE组RAW264.7细胞F4/80荧光强度较对照组明显增强,而4HNE+KL组F4/80荧光强度较4HNE组明显减弱(均P<0.05)。CCK8实验结果显示,相比于Mø-NC组,Mø-4HNE组HUVECs增生显著增加,而Mø-4HNE+KL组HUVECs增生较Mø-4HNE组显著下降(均P<0.01)。划痕实验和Transwell实验结果显示,相比于Mø-NC组,Mø-4HNE组HUVECs迁移显著增强,而Mø-4HNE+KL组HUVECs迁移较Mø-4HNE组显著减弱(均P<0.01)。管腔形成实验结果显示,相比于Mø-NC组,Mø-4HNE组HUVECs管腔数显著增加,而Mø-4HNE+KL组管腔数较Mø-4HNE组显著下降(均P<0.01)。Western blot实验结果显示,相比于 Mø-NC组,Mø-4HNE组HUVECs中Claudin 5、Occludin、ZO 1蛋白的相对表达量明显减少,而Mø-4HNE+KL组Claudin 5、Occludin、ZO 1蛋白的相对表达量较Mø-4HNE组明显增加(均P<0.01)。

结论：KL通过改变糖尿病氧化应激环境中巨噬细胞激活状态抑制了HUVECs的增生、迁移、管腔形成,并增强了HUVECs的紧密连接。     

GC法测定小儿氨酚烷胺颗粒中盐酸金刚烷胺

目的建立GC法测定小儿氨酚烷胺颗粒中盐酸金刚烷胺。方法采用HP-5毛细管柱（30 m×0.32 mm×0.25μm）,FID检测器;气化室温度为220℃,检测室温度为300℃。采用程序升温,初始温度为70℃,以10℃/min升至140℃。载气：氮气,流速为4 mL/min;进样量为1μL;分流比：1∶50。采用内标法定量。结果盐酸金刚烷胺在0.1~0.7 mg/mL线性关系良好（r=0.999 6）。平均回收率为99.27%,RSD值为0.66%（n=9）。结论本法简便、准确、重复性好,可用于小儿氨酚烷胺颗粒中盐酸金刚烷胺的测定。     

电缆附件安装操作技能对安装质量的影响（3）

本文对安装中运用刀具不当所产生的负面影响和电缆附件安装中安装工艺的要点作了简要分析。     

基于UPLC/Q-TOF MS/MS的稳心颗粒主要成分分析及鉴定

[目的]采用超高效液相色谱-四级杆-飞行时间质谱(UPLC/Q-TOF MS/MS)技术,对稳心颗粒中的化学成分进行初步分析。[方法]采用ACQUITY UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7μm),以乙腈-水(含体积比0.1%甲酸)为流动相梯度洗脱,流速为0.3 mL/min,电喷雾电离源,正/负离子双模式检测,通过UPLC/Q-TOF MS/MS所得到的化合物分子量以及MS/MS碰撞解离碎片,结合文献报道,对稳心颗粒的主要化学成分进行分析鉴定。[结果]结合文献报道和一级MS、二级MS/MS高分辨质谱数据,对其中68个化合物进行了初步鉴定,结果显示,稳心颗粒成分以糖类、皂苷类、黄酮类为主,其中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1丰度较高。[结论] UPLC/Q-TOF MS/MS技术是鉴定中药复方成分的有效手段,本研究首次利用该技术对稳心颗粒的主要成分进行定性分析,为阐明该复方药效物质基础、探讨其作用机制及其质量控制提供了有效的科学依据。     

灌注液中庆大霉素对白内障患者结膜囊细菌的作用

目的:分析白内障手术灌注液中庆大霉素对白内障患者结膜囊细菌的作用,为制定围手术期眼内炎预防措施提供依据。方法:采集200例白内障患者结膜囊拭子标本,进行细菌培养鉴定和药物敏感试验,观察检出细菌对20 μg/mL、40 μg/mL、80 μg/mL庆大霉素和2 μg/mL左氧氟沙星敏感率。结果:结膜囊细菌培养阳性检出率为62%,其中以表皮葡萄球菌为主,耐甲氧西林金黄色葡萄球菌（MRSA）检出率为3.22%,耐甲氧西林表皮葡萄球菌（MRSE）检出率为28.2%;全部检出细菌对20 μg/mL、40 μg/mL、80 μg/mL庆大霉素和2 μg/mL左氧氟沙星敏感率分别为79.84%、95.16%、97.58%、78.23%,中高浓度庆大霉素和左氧氟沙星比较,差异有统计学意义（均P<0.05）;MRSA和MRSE对20 μg/mL、40 μg/mL、80 μg/mL庆大霉素和2 μg/mL左氧氟沙星敏感率分别为69.23%、94.87%、100%、41.03%,各浓度庆大霉素和左氧氟沙星比较,差异有统计学意义（均P<0.05）。结论:白内障患者结膜囊细菌对灌注液中庆大霉素具有较高的敏感度,灌注液中加入适宜浓度庆大霉素,作为白内障围手术期眼内炎预防的措施应继续加以推广。     

角膜后表面散光的评估方法及临床特征的研究进展

角膜散光的精准评估在屈光和白内障手术中具有重要的临床意义。以往由于理论认识的不足和检查设备的局限性,低估了角膜后表面散光（PCA）对整体角膜散光评估所带来的影响。近年来,基于不同原理的多种设备实现了对角膜后表面信息的直接或间接采集,其在临床和科研中的广泛应用使PCA的分布特点及规律逐渐清晰。现对PCA的评估方法和临床特征作一综述,旨在提高临床实践中对PCA的认识和重视。     

Human umbilical cord-derived mesenchymal stem cells treatment for refractory uveitis: a case series

AIM: To evaluate therapeutic outcomes of human umbilical cord-derived mesenchymal stem cells (HUC-MSCs) treatment in patients with refractory uveitis. METHODS: A retrospective and noncomparative review was performed on four patients with refractory uveitis from December 2013 to December 2017. HUC-MSCs were administered intravenously at a dose of 1×106 cells/kg. Clinical response, relapse rate, change of visual acuity, and other metrics were evaluated. RESULTS: All four patients presented with responses to HUC-MSCs treatment, with three males and one female. The numbers of uveitis attacks per year after the HUC-MSCs treatment (0, 2, 0, 0 respectively) all decreased compared with the numbers before the treatment (3, 6, 4, 4 respectively). The oral steroid and immunosuppressive agents were tapered in all patients without recrudescence of ocular inflammation, and three patients discontinued their oral medicine at the last visit. The best corrected visual acuity (BCVA) of 3 patients was improved to varying degrees, and the BCVA of 1 patient remained at 20/20 (Snellen chart) from the first to the last consultation. CONCLUSION: The study provides an effective therapy of HUC-MSCs in maintaining remission in patients affected by uveitis refractory to previous immunosuppressant treatments.     

Novel rhodamine dye with large Stokes shifts by fusing the 1,4-diethylpiperazine moiety and its applications in fast detection of Cu2+

Rhodamine dyes were widely developed for designing probes due to their excellent photophysical properties and biocompatibility. However, traditional rhodamine dyes still bear major drawbacks of short emission wavelengths (<600 nm) and narrow Stokes shifts (<30 nm), which limit their biological imaging applications. Herein, we reported a novel mitochondria-targeted fluorescent dye JRQ with near-infrared (NIR) emission wavelength and improved Stokes shift (63 nm) by tuning the donor–acceptor–donor (D–A–D) character of the rhodamine skeleton. As expected, JRQ exhibited multiple excellent properties and could accumulate in mitochondria, and can therefore be used as a signal reporter for the design of fluorescent probes by taking advantage of the fluorescence controlled mechanism of the ring opening and closing chemical processes of the spirolactone platform. By using JRQ as a precursor, a highly sensitive fluorescent probe JRQN for the fast detection of mitochondrial Cu²⁺ ions was synthesized based on the Cu²⁺-triggered specific hydrolysis mechanism because mitochondria are an important reservoir of intracellular Cu²⁺. We expect that the Stokes shift increase of rhodamine dyes via tuning the donor–acceptor–donor (D–A–D) character of the rhodamine skeleton will provide a novel synthetic approach for the development of rhodamine dyes and expansion of their applications.

A novel NIR rhodamine dye JRQ with large Stokes shift (70 nm) by fusing the 1,4-diethylpiperazine moiety in rhodamine dyes has been synthesized and utilized to design a probe.