Carboxylesterase catalyzed 18O-labeling of carboxylic acid and its potential application in LC-MS/MS based quantification of drug metabolites

LC-MS quantification of drug metabolites is sometimes impeded by the availability of internal standards that often requires customized synthesis and/or extensive purification. Although isotopically labeled internal standards are considered ideal for LC-MS/MS based quantification, de novo synthesis using costly isotope-enriched starting materials makes it impractical for early stage of drug discovery. Therefore, quick access to these isotope-enriched compounds without chemical derivatization and purification will greatly facilitate LC-MS/MS based quantification. Herein, we report a novel ¹⁸O-labeling technique using metabolizing enzyme carboxylesterase (CES) and its potential application in metabolites quantification study. Substrates of CES typically undergo a two-step oxygen exchange with H₂¹⁸O in the presence of the enzyme, generating singly- and doubly-¹⁸O-labeled carboxylic acids; however, unexpected hydrolytic behavior was observed for three of the test compounds – indomethacin, piperacillin and clopidogrel. These unusual observations led to the discovery of several novel hydrolytic mechanisms. Finally, when used as internal standard for LC-MS/MS based quantification, these in situ labeled compounds generated accurate quantitation comparable to the conventional standard curve method. The preliminary results suggest that this method has potential to eliminate laborious chemical synthesis of isotope-labeled internal standards for carboxylic acid-containing compounds, and can be developed to facilitate quantitative analysis in early-stage drug discovery.     

Insights from examination of hearts from adults dying suddenly to the understanding of congenital cardiac malformations

Congenital heart disease is a rare but important finding in adults who experience sudden death. Examination of the congenitally malformed heart has historically been considered esoteric and best left to those with expertise. The Cardiac Risk in the Young cardiovascular pathology laboratory based at St George's University of London has now received over 6,000 cases. Of these, 21 congenitally malformed hearts were retained for research and educational purposes. Hearts were assessed using sequential segmental analysis, and causes of death were adjudicated based on thorough macroscopic examination and histology. Congenital malformations that were encountered included atrial septal defects, ventricular septal defects, tetralogy of Fallot, and transposition of the great arteries in both its regular and congenitally corrected variants. Findings also included hearts with mirror-imaged and isomeric atrial appendages. Direct causes of death included myocardial fibrosis, pulmonary hypertension, and hemorrhage. A small but notable proportion did not reveal a substrate for arrhythmia, raising the question of whether the terminal event was due to the congenital heart disease itself, or an underlying channelopathy. Here, we demonstrate the value of simple sequential segmental analysis in describing and categorizing the cases, with the concept of the “morphological method” serving to identify the distinguishing features of the cardiac components. Clin. Anat. 33:394–404, 2020. © 2019 Wiley Periodicals, Inc.     

顶空气相色谱-质谱法测定布南色林原料药中遗传毒性杂质

目的：建立顶空气相色谱-质谱法测定布南色林原料药中甲磺酸甲酯、甲磺酸乙酯和甲磺酸异丙酯3种遗传毒性杂质含量的方法。方法：采用顶空气相色谱-质谱法，以甲磺酸丁酯为内标，按内标标准曲线法进行甲磺酸甲酯、甲磺酸乙酯和甲磺酸异丙酯的含量测定。色谱条件：DB-WAX毛细管色谱柱（30 m×0.25 mm×0.25 μm）；程序升温，初始柱温为40℃，维持3 min，升温速率为30℃·min^-1，终止温度150℃，保持2 min；进样口温度为110℃；载气（He）流速为0.6 mL·min^-1；进样量为1 mL；进样方式为分流进样，分流比为20：1。质谱条件：电子轰击离子源（EI），扫描方式为选择性离子检测；离子源温度为200℃；接口温度为150℃；电子能量为70 eV；溶剂延迟1 min。结果：3种杂质成分之间的分离度均大于2.0；甲磺酸甲酯、甲磺酸乙酯、甲磺酸异丙酯检测质量浓度线性范围均为0.025~3.0 μg·mL^-1（r ≥ 0.998 5）；精密度、稳定性、重复性试验的RSD<5%；加样回收率分别为93.40%~101.40%（RSD为3.2%，n=9）、92.80%~99.70%（RSD为2.5%，n=9）和96.30%~100.75%（RSD为1.6%，n=9）。结论：该方法简便、准确、灵敏、迅速，可用于布南色林原料药中3种遗传毒性杂质的测定。     

翻转课堂教学法在消化内科临床带教的应用评价

目的 探讨翻转课堂教学法在消化内科临床带教的教学效果.方法 选取2018年10月—2019年10月在消化内科进行临床见习的护生48人(全部为女生),按照实习先后顺序,前24人作为试验组,给予翻转课堂教学法教学;后24人作为对照组,采用传统的带教模式.讨论两组护生在理论和操作成绩和对带教满意度方面有无差异.两组学生在试验开始之前的成绩相比较,差异无统计学意义(P>0.05).结果 试验组在理论、操作、教学满意度等方面的成绩均高于对照组,P<0.05.结论 翻转课堂教学法在消化内科的临床带教中有较好的教学效果,可以提高学生的学习成绩和教学满意度.     

在手术室护理教学中应用目标教学法的效果观察

目的探讨目标教学法在手术室护理教学中的应用效果。方法选取2017年10月-2018年10月期间80名手术室的实习护生,按照完全随机对照原则分组,对照组(n=40)采用传统带教法,观察组(n=40)采用目标教学法,分析技能操作考核成绩、护生对教学方式的满意度评分以及教师对学生表现的满意度评分。结果观察组技能操作考核成绩、护生对教学方式的满意度评分以及教师对学生表现的满意度评分高于对照组,差异有统计学意义(P<0.05)。结论在手术室护理教学中应用目标教学法效果较好,有助于护生将理论与实践更好地结合,进而有效提高实习质量以及护生的临床思维能力。     

基于子宫内膜异位症与恶性肿瘤的相关性探析补肾化瘀法作用机制及中医理论依据

子宫内膜异位症其异位内膜组织虽然在形态学上呈良性表现，却具有类似恶性肿瘤的生物学特性。中医学一般认为，“正虚伏邪”为恶性肿瘤的病机特点；那么，肾虚血瘀既属于“正虚伏邪”的范畴，又体现了EMs发病学和疾病发生、发展过程中的主要特点。在“病证相应”的中医治则之下，补肾化瘀法的临床疗效主要表现为：缓解痛经症状、提高受孕率，以及调整月经周期。本文基于EMs与恶性肿瘤的相关性，通过分析补肾化瘀法治疗EMs的理论依据及其抑制异位内膜侵袭的调控机制，主要包括解除免疫抑制、阻断局部微血管新生等，旨在阐明其疗效显著的原因，为临床推广提供可靠的基础研究证据。     

经穹隆结膜入路埋线法重睑成形术应用探讨

目的:通过分析重睑形成原理,综合比较多种重睑成形的方法,讨论埋线法重睑成形术的优点和可行性。方法:对387例患者行经穹隆结膜入路间断缝合埋线法重睑术。其中88例为其他方法埋线后重睑线消失的患者。结果:手术方法可靠,术后所形成的重睑外形良好,重睑维持时间长。最长随访时间38个月。结论:此方法因符合重睑形成原理,可以充分的将较多的上睑提肌腱膜与上睑皮肤结扎固定、故术后所形成的重睑牢固。且在直视下经穹隆结膜入路,降低了对眼球组织损伤的危险性。是埋线法重睑成形术一种可靠的手术方法。     

启发式教学在生理学教学中的应用研究

针对生理学课程理论抽象、实验操作复杂等特点,阐述了启发式教学方法取代传统填鸭式教学方法的必然性。因生理学教学难度大、实验操作条件高,故着重介绍了问答式、讨论式、演绎式、归纳式、直观式等五种启发式教学方法在生理学教学中的应用。     

杆菌肽发酵的培养基优化及影响其检测的因素

通过对影响杆菌肽抑菌圈大小的各种因素的研究，从而得到提高地衣芽抱杆菌发酵杆菌肽的产量及测量的最佳效果。研究结果表明豆芽汁培养基发酵的杆菌肽产量最高，并且在豆芽汁中加糖量为2g／dL时，抑菌圈最大。管碟法测定时，培养基的厚度影响最为显著，下层厚度10mL，上层5mL最佳，上层培养基的酸碱度为7．0，指示菌的浓度在10^8cfu／mL时，抑菌圈最清晰，直径最大，平均直径为2．52cm。