BACKGROUND & AIMS: OBJECTIVES:To validate the SAPS 3 admission prognostic model in patients with cancer admitted to the intensive care unit (ICU). DESIGN:Cohort study. SETTING:Ten-bed medical-surgical oncologic ICU. PATIENTS AND PARTICIPANTS:Nine hundred and fifty-two consecutive patients admitted over a 3-year period. INTERVENTIONS:None. MEASUREMENTS AND RESULTS:Data were prospectively collected at admission of ICU. SAPS II and SAPS 3 scores with respective estimated mortality rates were calculated. Discrimination was assessed by area under receiver operating characteristic (AUROC) curves and calibration by Hosmer-Lemeshow goodness-of-fit test. The mean age was 58.3+/-23.1 years; there were 471 (49%) scheduled surgical, 348 (37%) medical and 133 (14%) emergency surgical patients. ICU and hospital mortality rates were 24.6% and 33.5%, respectively. The mean SAPS 3 and SAPS II scores were 52.3+/-18.5 points and 35.3+/-20.7 points, respectively. All prognostic models showed excellent discrimination (AUROC>or=0.8). The calibration of SAPS II was poor (p<0.001). However, the calibration of standard SAPS 3 and its customized equation for Central and South American (CSA) countries were appropriate (p>0.05). SAPS II and standard SAPS 3 prognostic models tended somewhat to underestimate the observed mortality (SMR>1). However, when the customized equation was used, the estimated mortality was closer to the observed mortality [SMR=0.95 (95% CI=0.84-1.07)]. Similar results were observed when scheduled surgical patients were excluded. CONCLUSIONS:The SAPS 3 admission prognostic model at ICU admission, in particular its customized equation for CSA, was accurate in our cohort of critically ill patients with cancer.

背景与目标：

BACKGROUND & AIMS: :Water plays an active role in many fundamental phenomena in cellular systems such as molecular recognition, folding and conformational equilibria, reaction kinetics and phase partitioning. Hence, our ability to account for the energetics of these processes is highly dependent on the models we use for calculating solvation effects. For example, theoretical prediction of protein-ligand binding modes (i.e., docking) and binding affinities (i.e., scoring) requires an accurate description of the change in hydration that accompanies solute binding. In this review, we discuss the challenges of constructing solvation models that capture these effects, with an emphasis on continuum models and on more recent developments in the field. In our discussion of methods, relatively greater attention will be given to boundary element solutions to the Poisson equation and to nonpolar solvation models, two areas that have become increasingly important but are likely to be less familiar to many readers. The other focus will be upon the trending efforts for evaluating solvation models in order to uncover limitations, biases, and potentially attractive directions for their improvement and applicability. The prospective and retrospective performance of a variety of solvation models in the SAMPL blind challenges will be discussed in detail. After just a few years, these benchmarking exercises have already had a tangible effect in guiding the improvement of solvation models.

背景与目标： : 水在细胞系统中的许多基本现象中起着积极作用，例如分子识别，折叠和构象平衡，反应动力学和相分配。因此，我们解释这些过程的能量学的能力在很大程度上取决于我们用于计算溶剂化效应的模型。例如，蛋白质-配体结合模式 (即对接) 和结合亲和力 (即评分) 的理论预测需要准确描述伴随溶质结合的水合变化。在这篇综述中，我们讨论了构建捕获这些影响的溶剂化模型的挑战，重点是连续模型和该领域的最新发展。在我们对方法的讨论中，将相对更多地关注泊松方程的边界元解和非极性溶剂化模型，这两个领域已变得越来越重要，但许多读者可能不太熟悉。另一个重点将放在评估溶剂化模型的趋势上，以发现局限性，偏见以及其改进和适用性的潜在吸引力方向。将详细讨论SAMPL盲挑战中各种溶剂化模型的前瞻性和回顾性表现。短短几年后，这些基准测试工作已经在指导溶剂化模型的改进方面产生了切实的效果。

BACKGROUND & AIMS: BACKGROUND:Oral health literacy is a newly emerging field with considerable research potential. AIM:To validate an original instrument, the Hong Kong Oral Health Literacy Assessment Task (HKOHLAT-P) for paediatric dentistry. DESIGN:A convenient sample of 200 child/parent dyads attending a dental hospital in Hong Kong was selected. Convergent validity was tested by examining the association of HKOHLAT-P scores with those derived from the Test of Functional Health Literacy in Dentistry (TOFHLiD) and Hong Kong Rapid Estimate of Adult Literacy in Dentistry (HKREALD-30). The predictive validity of HKOHLAT-P was determined by testing the association between HKOHLAT-P and children's caries experience (dmft) and the Chinese Early Childhood Oral Health Impact Scale (ECOHIS). The test-retest reliability and internal consistency of HKOHLAT-P were also evaluated. RESULTS:HKOHLAT-P was positively correlated with TOFHLiD and HKREALD-30 (P < 0.01), and was negatively correlated with children's dmft and ECOHIS. In the regression model, HKOHLAT-P was associated with TOFHLiD, HKEALD-30, children's dmft, and ECOHIS (P < 0.05) after controlling for participants' demographic characteristics. The intra-class correlation coefficient of HKOHLAT-P was 0.63 and the Cronbach's α was 0.71. CONCLUSION:Initial testing of HKOHLAT-P suggested that it is a valid and reliable instrument.

背景与目标：

BACKGROUND & AIMS: AIMS:To reduce sudden cardiac death, implantable cardioverter-defibrillators (ICDs) are indicated in patients with ischaemic and non-ischaemic dilated cardiomyopathy and a left ventricular ejection fraction (LVEF) ≤35%. Current guidelines do not recommend device therapy in patients with a life expectancy <1 year since benefit in these patients is low. In this study, we evaluated the incidence and predictors of early mortality (<1 year after implantation) in a consecutive primary prevention population. METHODS AND RESULTS:Analysis was performed on a prediction and validation cohort. The primary endpoint was all-cause mortality at 1 year. The prediction cohort comprised 861 prophylactic ICD recipients with ischaemic cardiomyopathy or dilated cardiomyopathy from the Academic Medical Center (Amsterdam) and Thorax Center Twente (Enschede). Detailed clinical data were collected. After multivariate analysis, a risk score was developed based on age ≥75 years, LVEF ≤ 20%, history of atrial fibrillation, and estimated glomerular filtration rate (eGFR) ≤30 mL/min/1.73 m(2). Using these predictors, a low (≤1 factor), intermediate (2 factors), and high (≥3 factors) risk group could be identified with 1-year mortality of, respectively, 3.4, 10.9, and 38.9% (P< 0.01). Afterwards, the risk score was validated in 706 primary prevention patients from the Erasmus Medical Center (Rotterdam). One-year mortality was, respectively, 2.5, 13.2, and 46.3% (all P< 0.01). CONCLUSION:A simple risk score based on age, LVEF, eGFR, and atrial fibrillation can identify patients at low, intermediate, and high risk for early mortality after ICD implantation. This may be helpful in the risk assessment of ICD candidates.

背景与目标：

BACKGROUND & AIMS: :Successful translation of findings derived from preclinical studies into effective therapies is critical in biomedical research. Lack of robustness and reproducibility of the preclinical data, due to insufficient number of repeats, inadequate cell-based and mouse models contribute to the poor success rate. Antibodies are widely used in preclinical research, notably to determine the expression of potential therapeutic targets in tissues of interest, including tumors, but also to identify disease and/or treatment response biomarkers. We sought to determine whether the current antibody characterization standards in preclinical research are sufficient to ensure reliability of the data found in peer-reviewed publications. To address this issue, we used detection of the protein c-FLIP, a major factor of resistance to apoptosis, as a proof of concept. Accurate detection of endogenous c-FLIP levels in the preclinical settings is imperative since it is considered as a potential theranostic biomarker. Several sources of c-FLIP antibodies validated by their manufacturer and recommended for western blotting were therefore rigorously tested. We found a wide divergence in immune recognition properties. While these antibodies have been used in many publications, our results show that several of them failed to detect endogenous c-FLIP protein by Western blotting. Our results suggest that antibody validation standards are inadequate, and that systematic use of genetic knockdowns and/or knockouts to establish proof of specificity is critical, even for antibodies previously used in the scientific literature. Because antibodies are fundamental tools in both preclinical and clinical research, ensuring their specificity is crucial.

背景与目标： : 成功地将临床前研究的发现转化为有效的疗法在生物医学研究中至关重要。由于重复次数不足，基于细胞的和小鼠模型不足，缺乏临床前数据的稳健性和可重复性导致成功率低。抗体广泛用于临床前研究，特别是用于确定感兴趣组织 (包括肿瘤) 中潜在治疗靶标的表达，还用于鉴定疾病和/或治疗反应生物标志物。我们试图确定临床前研究中当前的抗体表征标准是否足以确保同行评审出版物中发现的数据的可靠性。为了解决这个问题，我们使用了检测蛋白c-FLIP (抗凋亡的主要因素) 作为概念的证明。临床前环境中内源性c-FLIP水平的准确检测势在必行，因为它被认为是潜在的肿瘤生物标志物。因此，对其制造商验证并推荐用于蛋白质印迹的c-FLIP抗体的几种来源进行了严格测试。我们发现免疫识别特性存在很大差异。尽管这些抗体已在许多出版物中使用，但我们的结果表明，其中一些未能通过Western印迹检测到内源性c-FLIP蛋白。我们的结果表明，抗体验证标准是不充分的，并且系统地使用基因敲除和/或敲除来建立特异性证明是至关重要的，即使对于以前在科学文献中使用的抗体也是如此。由于抗体是临床前和临床研究的基本工具，因此确保其特异性至关重要。

BACKGROUND & AIMS: :The strong link between gene expression of mitotic Aurora kinases and cancer has stimulated a very high interest in developing Aurora kinase inhibitors for cancer therapy. Validation of Aurora kinases as targets, and development of pharmacodynamic biomarkers for inhibitors of Aurora kinases, provides an example of how target validation can help the drug discovery process, and also of how to interpret results depending on the technology used. In this review, we outline the principal tools, concepts, and strategies of target and biomarker validation for Aurora kinases, with emphasis on validation results derived from RNA-interference experiments. These data were essential for the decision to enter the next steps in drug development and for the selection of the appropriate biomarkers for clinical trials.

背景与目标： : 有丝分裂极光激酶的基因表达与癌症之间的紧密联系激发了人们对开发用于癌症治疗的极光激酶抑制剂的高度兴趣。Aurora激酶作为靶标的验证以及Aurora激酶抑制剂的药效学生物标志物的开发，提供了靶标验证如何帮助药物发现过程以及如何根据所使用的技术解释结果的示例。在这篇综述中，我们概述了Aurora激酶的靶标和生物标志物验证的主要工具，概念和策略，重点是RNA干扰实验得出的验证结果。这些数据对于决定进入药物开发的下一步以及为临床试验选择合适的生物标志物至关重要。

BACKGROUND & AIMS: BACKGROUND:Little research has been conducted to validate pain assessment tools in critical care, especially for patients who cannot communicate verbally. OBJECTIVE:To validate the Critical-Care Pain Observation Tool. METHODS:A total of 105 cardiac surgery patients in the intensive care unit, recruited in a cardiology health center in Quebec, Canada, participated in the study. Following surgery, 33 of the 105 were evaluated while unconscious and intubated and 99 while conscious and intubated; all 105 were evaluated after extubation. For each of the 3 testing periods, patients were evaluated by using the Critical-Care Pain Observation Tool at rest, during a nociceptive procedure (positioning), and 20 minutes after the procedure, for a total of 9 assessments. Each patient's self-report of pain was obtained while the patient was conscious and intubated and after extubation. RESULTS:The reliability and validity of the Critical-Care Pain Observation Tool were acceptable. Interrater reliability was supported by moderate to high weighted kappa coefficients. For criterion validity, significant associations were found between the patients' self-reports of pain and the scores on the Critical-Care Pain Observation Tool. Discriminant validity was supported by higher scores during positioning (a nociceptive procedure) versus at rest. CONCLUSIONS:The Critical-Care Pain Observation Tool showed that no matter their level of consciousness, critically ill adult patients react to a noxious stimulus by expressing different behaviors that may be associated with pain. Therefore, the tool could be used to assess the effect of various measures for the management of pain.

背景与目标：

BACKGROUND & AIMS: :The aim of the present study was to test the influence of obesity and the presence of type 2 diabetes mellitus (T2DM) on the expression of ten housekeeping genes and of the 18S rRNA in a group of human adipose tissue samples from the omental and subcutaneous depot. Adipose tissue biopsies were obtained by laparoscopic surgery from lean and obese patients. After the extraction, mRNA levels in adipose tissue samples were quantified by real-time PCR using the commercial HUMAN ENDOGENOUS CONTROL PLATES. From the genes analyzed, 18S rRNA exhibited the most stable expression levels in both depots regardless of the pathophysiological conditions of obesity and obesity-associated T2DM. Contrarily, GAPD was the gene with the highest variation in its expression levels, being upregulated (8.0-fold) in the obese group and downregulated (3.5-fold) in obesity-associated T2DM. Our results show that 18S rRNA may be the most suitable gene for normalization in expression studies performed in human adipose tissue samples obtained from patients suffering from obesity and/or obesity-associated T2DM, whereas GAPD is less appropriate for comparison purposes under these circumstances.

背景与目标： : 本研究的目的是测试肥胖和2型糖尿病 (T2DM) 的存在对来自网膜和皮下仓库的一组人类脂肪组织样品中十个管家基因和18S rRNA表达的影响。脂肪组织活检是通过腹腔镜手术从瘦和肥胖患者中获得的。提取后，使用商业人类内源性对照板通过实时PCR定量脂肪组织样品中的mRNA水平。从分析的基因来看，无论肥胖和肥胖相关的T2DM的病理生理状况如何，18S rRNA在两个仓库中均表现出最稳定的表达水平。相反，GAPD是其表达水平变化最高的基因，在肥胖组中上调 (8.0倍)，在肥胖相关的T2DM中下调 (3.5倍)。我们的结果表明，在从肥胖和/或肥胖相关的T2DM患者获得的人脂肪组织样本中进行的表达研究中，18S rRNA可能是最适合标准化的基因，而在这些情况下，GAPD不太适合用于比较目的。

BACKGROUND & AIMS: UNLABELLED:This article summarizes the work done to adapt and to validate the short form of Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12) for its use in Spain. It will become the first validated questionnaire in this country for the evaluation of the sexual function in women with Pelvic Organ Prolapse and/or Urinary Incontinence. PATIENTS AND METHOD:49 women who visited a specialized unit with symptoms of pelvic floor were included. Patients filled in the Spanish version of the questionnaire to validate (PISQ-12), the Urinary Incontinence Questionnaire (ICIQ-UI-SF); the Female Sexual Function Questionnaire (FSM) and the Bladder Control Autoevaluation Questionnaire (CACV). Factibility, reliability and validity of the new questionnaire were evaluated. RESULTS:Factibility: 99.83% of the sample answered all the items (only one patient did not answer one of the items); average administration time 3.5 (1.5) minutes. RELIABILITY:Cronbach's alpha was 0,829. VALIDITY:PISQ-12 correlation with FSM was 0,71; with ICIQ-UI-SF it was -0,038; with the CACV "symptoms" dimension the correlation was -0,30 and with the "discomfort" dimension it was -0,40. The existence of the same three dimensions of the PISQ-12 original version in the adapted Spanish questionnaire is checked through a factorial analysis. The score in PISQ-12 was worse (lower) in the case of women with Hyperactive Bladder symptoms and discomfort measured with the CACV questionnaire and in women with sexual dysfunction measured with FSM. PISQ-12 is an instrument with the appropriate psychometric characteristics to evaluate sexual function in women with pelvic floor problems.

背景与目标：

BACKGROUND & AIMS: BACKGROUND:Traffic Injuries are a major public health problem, especially among young people. However, we have not found any useful questionnaire designed in our country for the epidemiological research in this field. The objective of this study was to design and validate an easy and quickly-to-fill questionnaire aimed to collect information on how frequently university car drivers report to be involved in driving circumstances theoretically related to traffic crashes. METHODS:Between 2007 and 2010, a total of 1597 young undergraduate students at the University of Granada answered a self-administered questionnaire collecting information about exposure, accidents and involvement in 28 different driving circumstances. For designing this questionnaire, an extensive literature review was carried out and the opinions of five experts in a panel were also taken into account. By applying the tetracoric correlation coefficient, we conducted a factor analysis. Internal consistency was assessed using Cronbach's alpha coefficient. Finally, we evaluated the crude and adjusted association of each identified factor with the odds for having suffered an accident. RESULTS:After excluding 8 circumstances, the remaining ones were grouped into three factors: the first one included ten high-prevalence circumstances and explained 31.9% of the total variability. Meanwhile, the other two factors included five circumstances each one which respectively explained 15.2% and 12.5% of the variability. Cronbach's alpha coefficients ranged between 0.816 and 0.553. When adjustments according age, sex, years in possession of the driving license and intensity of exposure were made, the first factor obtained the score more strongly associated with the accident rate (OR = 1.51; CI95%: 1.25-1.85). CONCLUSIONS:The final version (20 circumstances) identified three factors related to higher accident rates among the young drivers. The first one integrated, among other circumstances, the excessive speed and driving while sleepy or tired and it was the most closely associated with the accident rate in the adjusted analysis. The second factor included, among others, the commission of driving offences, and the third one included driving under the influence of alcohol, not always wearing the seat belt and distractions.

背景与目标：

BACKGROUND & AIMS: PURPOSE:The present study validated the abbreviated version of the World Health Organization Quality of Life (WHOQOL-BREF) questionnaire for general use in New Zealand. METHODS:A random postal sample from the national electoral roll was used, and 808 questionnaires were returned. Psychometric properties of the instrument were assessed, including tests of the four-domain factor structure using confirmatory factor analysis and Rasch analysis. RESULTS:Goodness-of-fit from the confirmatory factor analysis were good, and the overall conclusion of the Rasch analysis supported the confirmatory factor analysis (CFA) findings after dealing with problems of threshold ordering, local dependency, and differential item functioning (DIF). CONCLUSIONS:The WHOQOL-BREF is valid for general use in New Zealand. In the future work, the WHOQOL-BREF domain scores should either be analyzed using non-parametric statistics or data should be fitted to the Rasch model to derive interval person estimates.

背景与目标：

BACKGROUND & AIMS: :In this review we have showcased the preclinical development of original amphiphilic nanomaterials designed for ruthenium-based anticancer treatments, to be placed within the current metallodrugs approach leading over the past decade to advanced multitarget agents endowed with limited toxicity and resistance. This strategy could allow for new options for breast cancer (BC) interventions, including the triple-negative subtype (TNBC) with poor therapeutic alternatives. BC is currently the second most widespread cancer and the primary cause of cancer death in women. Hence, the availability of novel chemotherapeutic weapons is a basic requirement to fight BC subtypes. Anticancer drugs based on ruthenium are among the most explored and advanced next-generation metallotherapeutics, with NAMI-A and KP1019 as two iconic ruthenium complexes having undergone clinical trials. In addition, many nanomaterial Ru complexes have been recently conceived and developed into anticancer drugs demonstrating attractive properties. In this field, we focused on the evaluation of a Ru(III) complex-named AziRu-incorporated into a suite of both zwitterionic and cationic nucleolipid nanosystems, which proved to be very effective for the in vivo targeting of breast cancer cells (BBC). Mechanisms of action have been widely explored in the context of preclinical evaluations in vitro, highlighting a multitarget action on cell death pathways which are typically deregulated in neoplasms onset and progression. Moreover, being AziRu inspired by the well-known NAMI-A complex, information on non-nanostructured Ru-based anticancer agents have been included in a precise manner.

背景与目标： : 在这篇综述中，我们展示了为基于钌的抗癌治疗而设计的原始两亲性纳米材料的临床前发展，这些材料被置于当前的金属药物方法中，该方法在过去十年中领先于具有有限毒性和抗性的先进多靶点药物。该策略可以为乳腺癌 (BC) 干预提供新的选择，包括治疗方案较差的三阴性亚型 (TNBC)。卑诗省目前是第二广泛的癌症，也是女性癌症死亡的主要原因。因此，新型化学治疗武器的可用性是对抗BC亚型的基本要求。基于钌的抗癌药物是最具探索和先进的下一代金属治疗药物之一，NAMI-A和KP1019作为两种标志性的钌配合物已经进行了临床试验。此外，最近已经构思了许多纳米材料Ru配合物，并将其开发成具有吸引力的抗癌药物。在这一领域，我们专注于评估一种Ru(III) 复合物-名为AziRu-掺入到一组两性离子和阳离子核脂纳米系统中，这被证明对体内靶向乳腺癌细胞非常有效 (BBC)。在体外临床前评估的背景下，已广泛探索了作用机制，强调了对细胞死亡途径的多靶点作用，这些作用通常在肿瘤的发作和进展中被解除管制。此外，由于AziRu受到著名的NAMI-A复合物的启发，有关非纳米结构的Ru基抗癌剂的信息已以精确的方式包含在内。

BACKGROUND & AIMS: :Screening is recommended to reduce both incidence and mortality of colorectal cancer. Currently, many countries employ fecal occult blood test (FOBT). In Emilia-Romagna (Italy), since 2005, FOBT immunochemical version (FIT) is performed every two years on people aged between 50 and 69 years. A colonoscopy is then carried out on those who are FIT positive. However, FIT shows approximately 65% false positives (non-tumoral bleedings), leading to many negative colonoscopies. The use of an economic and easy-to-use method to check FOBT-positives will improve screening effectiveness, reducing costs to the national health service. This work illustrates the results of a three-year clinical validation protocol (started in 2016) of a patented device composed of a core of nanostructured gas sensors. This device was designed to identify CRC presence by fecal volatile compounds, with a non-invasive, in vitro and low-cost analysis. Feces are, in fact, affected by tumor-volatile biomarkers, produced by cellular peroxidation and metabolic alterations. The protocol consisted in the analysis of fecal samples of FIT-positive subjects, using colonoscopy as a gold standard. A total of 398 samples were analyzed with machine learning techniques, leading to a sensitivity and specificity of 84.1% and 82.4%, respectively, and a positive predictive value of 72% (25-35% for FIT).

背景与目标： : 建议进行筛查以降低结直肠癌的发病率和死亡率。目前，许多国家/地区都采用粪便潜血测试 (FOBT)。2005年，在意大利的艾米利亚-罗马涅 (Emilia-Romagna)，fott免疫化学版本 (FIT) 每两年对年龄在50至69岁之间的人进行一次。然后对那些健康阳性的人进行结肠镜检查。然而，FIT显示大约65% 个假阳性 (非肿瘤出血)，导致许多阴性结肠镜检查。使用一种经济且易于使用的方法来检查FOBT阳性将提高筛查效果，从而降低国家卫生局的成本。这项工作说明了由纳米结构气体传感器核心组成的专利设备的三年临床验证协议 (2016年开始) 的结果。该设备旨在通过非侵入性，体外和低成本分析来识别粪便挥发性化合物是否存在CRC。实际上，粪便受细胞过氧化和代谢改变产生的肿瘤挥发性生物标志物的影响。该方案包括使用结肠镜检查作为金标准，对FIT阳性受试者的粪便样本进行分析。使用机器学习技术分析了总共398个样品，分别得出84.1% 和82.4% 的敏感性和特异性，阳性预测值为72% (FIT为25-35%)。

BACKGROUND & AIMS: :Clinical decision support systems are a combination of software techniques to help the clinicians in their medical decision making process via functionalities ranging from basic signal analysis to therapeutic planning and computerized guidelines. The algorithms providing all these functionalities must be very carefully validated on real patient data and must be confronted to everyday clinical practice. One of the main problems when developing these techniques is the difficulty to obtain high-quality complete patient records, comprising data coming both from the biomedical equipment (high-frequency signals), and from numerous other sources (therapeutics, imagery, clinical actions, etc.). In this paper, we present an infrastructure for developing and testing such software algorithms. It is based on a bedside workstation where testing different algorithms simultaneously on real-time data is possible in the ward. It is completed by a collaborative portal enabling different teams to test their software algorithms on the same patient records, making comparisons and cross-validations more easily.

背景与目标： : 临床决策支持系统是软件技术的组合，可通过从基本信号分析到治疗计划和计算机化指南的功能来帮助临床医生进行医疗决策。提供所有这些功能的算法必须在真实的患者数据上进行非常仔细的验证，并且必须面对日常临床实践。开发这些技术时的主要问题之一是难以获得高质量的完整患者记录，包括来自生物医学设备 (高频信号) 和来自许多其他来源 (治疗，图像，临床动作等) 的数据。在本文中，我们介绍了用于开发和测试此类软件算法的基础结构。它基于床边工作站，可以在病房中同时测试实时数据的不同算法。它由一个协作门户网站完成，使不同的团队能够在相同的患者记录上测试他们的软件算法，从而更容易进行比较和交叉验证。

BACKGROUND & AIMS: :Menadione (2-methyl-1,4-naphthoquinone, MQ), a component of multivitamin drugs with antihemorrhagic, antineoplastic, and antimalarial activity, is frequently used to investigate quinone-induced cytotoxicity. The formation of MQ conjugates with glutathione (GSH) by Michael addition and subsequent biotransformation to yield N-acetyl-l-cysteine conjugates is believed to be an important detoxification process. However, the resulting conjugates, 2-methyl-3-(glutathione-S-yl)-1,4-naphthoquinone (MQ-GS) and 2-methyl-3-(N-acetyl-l-cysteine-S-yl)-1,4-naphthoquinone (MQ-NAC), retain the ability to redox cycle and to arylate cellular nucleophiles. Although the nephrotoxicity and hepatotoxicity of MQ-thiol conjugates have been reported in vitro, methods for their determination in vivo have yet to be published. Herein, a highly sensitive, simple, and selective HPLC-chemiluminescence (HPLC-CL) coupled method is reported, allowing for the first time the simultaneous determination of MQ, MQ-GS, and MQ-NAC in rat plasma after MQ administration. Our method exploits the unique redox characteristics of MQ, MQ-GS, and MQ-NAC to react with dithiothreitol (DTT) to liberate reactive oxygen species (ROS) which are detected by a CL assay using luminol as a CL probe. To verify the proposed mechanism, MQ-GS and MQ-NAC were synthetically prepared. Specimen preparation involved solid-phase extraction on an Oasis HLB cartridge followed by isocratic elution on an ODS column. No interference from endogenous substances was detected. Linearity was observed in the range of 5-120 nM for MQ-GS and MQ-NAC and 10-240 nM for MQ, with detection limits (S/N of 3) of 1.4, 0.8, and 128 fmol for MQ-GS, MQ-NAC, and MQ, respectively. The application of our method reported here is the first to extensively study the stability and reversibility of thiol-quinones.

背景与目标： : 甲萘醌 (2-甲基-1,4-萘醌，MQ) 是具有抗出血，抗肿瘤和抗疟活性的多种维生素药物的成分，通常用于研究醌诱导的细胞毒性。通过Michael加成形成MQ缀合物与谷胱甘肽 (GSH) 并随后进行生物转化以产生N-乙酰基-l-半胱氨酸缀合物被认为是重要的解毒过程。然而，得到的共轭物，2-甲基-3-(谷胱甘肽-S-基)-1,4-萘醌 (mq-gs) 和2-甲基-3-(N-乙酰基-l-半胱氨酸-S-基)-1,4-萘醌 (mq-nac)，保留氧化还原循环和芳化细胞亲核试剂的能力。尽管已在体外报道了MQ-硫醇偶联物的肾毒性和肝毒性，但体内测定方法尚未公布。本文报道了一种高度灵敏、简单和选择性的HPLC-化学发光 (hplc-cl) 偶联方法，首次允许在MQ给药后同时测定大鼠血浆中的MQ、mq-gs和mq-nac。我们的方法利用MQ，mq-gs和mq-nac的独特氧化还原特性与二硫苏糖醇 (DTT) 反应释放活性氧 (ROS)，使用鲁米诺作为CL探针通过CL测定法检测到。为了验证所提出的机制，综合制备了mq-gs和mq-nac。标本制备涉及在Oasis HLB柱上进行固相萃取，然后在ODS柱上进行等度洗脱。未检测到内源性物质的干扰。对于mq-gs和mq-nac，在5-120 nM和10-240 nM范围内观察到线性，对于mq-gs、mq-nac和MQ，检出限 (S/N为3) 分别为1.4、0.8和128 fmol。本文报道的方法的应用是首次广泛研究硫醇-醌的稳定性和可逆性。