[Ru(dcbpy)3]2+ functionalized 2D/3D hybrid metal-organic frameworks (Ru-BTC-MOFs), which performed good ECL performance, were synthesized by on one-step cathodic electrodeposition with Zn(NO3)2 as metal ion source, Ru(dcbpy)3Cl2) (tri(4,4-dicarboxybipyridine) ruthenium chloride) and H3BTC (1,3,5-phenyl tricarboxylic acid) as organic ligand. The effect of electrodeposition voltage, electrodeposition time, H3BTC and [Ru(dcbpy)3]Cl2 concentration on the morphology and ECL performance of Ru-BTC-MOFs were investigated in detail. An ECL biosensor platform for sensitive detection of carcinoembryonic antigen (CEA) was constructed by further modified AuNPs and aptamer on the Ru-BTC-MOFs/GCE. The specifically recognition and adsorption of CEA would hinder the electron transfer between Ru-BTC-MOFs and trinpropylamine (TPA) as the co-reactive agent, resulting in decreasing the ECL intensity. Under the optimal experimental conditions, the ECL intensity and logarithm of CEA concentration showed a good linear relationship in the concentration range of 10.0 fg/mL to 10.0 ng/mL, and the detection limit was 5.56 fg/mL. The proposed sensor performed acceptable reproducibility, repeatability, anti-interference ability and stability, and it would be expected to have broad application prospects in clinical trials.

Synthetic 4-Chloroethcathinone (4-CEC) is a derivative of cathinone that belongs to one of the more severe abused substances among new psychoactive substances (NPS). Current researches on 4-CEC mainly focus on metabolite identification studies, and there is a lack of researches on pharmacokinetic, tissue distribution and metabolomics studies in vivo. A sensitive and reliable LC-MS/MS assay was developed and validated for the determination of 4-CEC concentrations in plasma and tissue homogenates. According to the pharmacokinetic results, the absorption and elimination of 4-CEC were faster after administration. The Cmax was 1896 ± 876 ng/ml, the peak time Tmax was 10.1 ± 9.2 min, and the elimination half-life t1/2 was 100.4 min. Metabolomics studies showed that the highest concentrations of 4-CEC were found in brain, lung, kidney and liver. The results of tissue biopsy showed that the liver, kidney and brain tissue had a certain degree of damage. After 4-CEC administration, amino acid-related metabolism and biosynthesis, lipid metabolism, niacin and niacinamide metabolism in mice were interfered, suggesting that 4-CEC could cause energy metabolism disorder in mice. The metabolic pathways and toxicity mechanisms related to 4-CEC entry into the body were explained at the overall metabolic level by multivariate data analysis, screening and identification of differential metabolites and metabolic pathway analysis.

咖啡酸、丹参素等小分子酚酸类成分有广泛的药理活性,丹参酚酸类成分在复方丹参制剂中发挥着主要的药效作用.据报道,小分子酚酸类成分在一定条件下发生自氧化,可引发自由基链反应.自由基还可通过复杂的机制偶联生成多种聚合物.其中,含1,4-苯并二烷结构的聚合物具有抗炎、抗肿瘤和抗病毒等药理活性.笔者对咖啡酸等小分子酚酸类成分的自氧化及自由基聚合机制、产物及药理活性进行综述,以期为阐明含小分子酚酸类成分相关产品的物质基础提供依据,为含1,4-苯并二烷结构的物质在化合物筛选、新药开发领域的应用提供参考.

预测药物在血浆中的蛋白结合率,有助于了解药物的药代动力学特征,对药物发现的早期研究有重要的参考价值.本研究收集了 2452个临床药物的血浆蛋白结合率信息,用Molecular Operating Environment(MOE)和Mordred两种软件计算分子描述符,将算得的分子描述符作为模型的输入特征.使用极端梯度提升(extreme gradient boosting,XGBoost)算法和随机森林(randomforest,RF)算法构建机器学习模型.结果表明,与MOE相比,将Mordred计算的分子描述符作为模型的输入,构建的模型预测性能更优.使用XGBoost算法和RF算法构建模型的预测性能结果相近,最优模型的R2均为0.715.此外,根据研究结果得出药物血浆蛋白结合率与药物分子的一些理化性质参数,如水溶性,辛醇/水分配系数以及共轭双键密切相关.通过这些参数预测药物血浆蛋白结合率具有方便快捷的优点,可以为相关药代动力学研究提供参考依据.

Acute liver failure (ALF) is a serious clinical disorder with high fatality rates. Mahuang decoction (MHD), a well-known traditional Chinese medicine, has multiple pharmacological effects, such as anti-inflammation, anti-allergy, anti-asthma, and anti-hyperglycemia. In this study, we investigated the protective effect of MHD against ALF. In the lipopolysaccharide and D-galactosamine (LPS/D-GalN)-induced ALF mouse model, the elevated activities of the serum alanine and aspartate transaminases as well as the liver pathological damage were markedly alleviated by MHD. Subsequently, a metabolomics study based on the ultrahigh performance liquid chromatograph coupled with Q Exactive Orbitrap mass spectrometry was carried to clarify the therapeutic mechanisms of MHD against ALF. A total of 36 metabolites contributing to LPS/D-GalN-induced ALF were identified in the serum samples, among which the abnormalities of 27 metabolites were ameliorated by MHD. The analysis of metabolic pathways revealed that the therapeutic effects of MHD are likely due to the modulation of the metabolic disorders of tricarboxylic acid (TCA) cycle, retinol metabolism, tryptophan metabolism, arginine and proline metabolism, nicotinate and nicotinamide metabolism, phenylalanine metabolism, phenylalanine, tyrosine and tryptophan synthesis, as well as cysteine and methionine metabolism. This study demonstrated for the first time that MHD exerted an obvious protective effect against ALF mainly through the regulation of TCA cycle and amino acid metabolism, highlighting the importance of metabolomics to investigate the drug-targeted metabolic pathways.

目的 建立基于机器学习的化合物血脑屏障通透性(logBB)预测模型,并评价其预测效果.方法 收集360个化合物的分子结构信息与相应的logBB,使用遗传算法结合极端梯度提升算法来构建化合物logBB预测模型.结果 经十折交叉验证,模型的预测平方相关系数为0.63,均方误差为0.23,表现出良好的预测性能.此外,通过对模型特征重要性的分析,总结了对化合物logBB影响程度最高的前5个分子描述符,分别是拓扑极性表面积、氢键、疏水/亲水性、油水分配系数、辛醇水分配系数.结论 模型的预测性能优于仅使用极端梯度提升算法或使用遗传算法结合支持向量机来构建的模型性能,且远优于discovery studio(DS,2016版)软件自带的预测算法性能.本研究可为治疗脑部相关疾病的药物研发提供指导性意义.

Hyperpigmentation is a common skin condition with serious psychosocial consequences. Decapeptide-12, a novel peptide, has been found to be safer than hydroquinone in reducing melanin content, with efficacy up to more than 50% upon 16 weeks of twice-daily treatment. However, the peptide suffers from limited transcutaneous penetration due to its hydrophilicity and high molecular weight. Therefore, decapeptide-12 was modified by adding a palmitate chain in an attempt to overcome this limitation. Molecular docking results showed that the two peptides exhibited similar biological activity towards tyrosinase. We also tested the effect of chemical penetration enhancers and microneedles to deliver the two peptides into and through skin, using an in vitro human skin permeation method. It was shown that the palm-peptide achieved the best skin retention owing to the increased lipophilicity. In addition, skin permeation of the palm-peptides was enhanced by the chemical skin penetration enhancers, namely, oleic acid and menthol. Skin permeation of the native peptide was enhanced by the microneedle patch but not the chemical skin penetration enhancers. Cutaneous absorption of the palm-peptides was estimated to have achieved its therapeutic concentration within skin. The combinatory approach of using molecular modification, chemical penetration enhancement, and microneedle patch proves to be useful to enhanceskin permeation of the peptides.

肝毒性是药物治疗过程中常见的副作用,也是导致新药临床试验失败以及药物撤市的主要原因.建立可以有效识别药物肝毒性的体外模型对于新药研发至关重要.传统的二维(2D)模型在培养过程中会逐渐丧失肝脏特异性功能,无法准确评价药物肝毒性.新兴的三维(3D)培养技术可模拟体内细胞微环境,有利于肝细胞的体外组装,从而使3D肝细胞模型表现出与体内肝细胞相似的表型以及药物毒性反应,在肝毒性评价方面具有很大潜力.

近年来,随着生物打印技术的进步,基于生物打印技术构建的体外模型用于新药筛选和毒性评价引起人们极大的关注.作为一种新型的组织工程技术,生物打印技术的核心是生物墨水,即装载不同细胞的生物材料.明胶基生物墨水广泛用于生物打印,打印的三维(3D)体外模型与传统的二维(2D)模型相比,具有明显的优越性.本文主要对基于明胶及其修饰产物的生物墨水用于药物筛选和毒性评价进行综述,并给出对新型明胶生物墨水的展望.

体内药物与毒物分析是一门运用药物分析原理和技术对生物机体中药物及其代谢物、内源性物质以及侵入生物体内的有毒物质进行定性和定量分析的应用学科.为了适应国家医药产业发展和禁毒大战略的布局对药物分析专业的需求,通过优化教学内容、改进教学方法、完善考核制度等途径,激发学生的学习兴趣,培养学生的创新能力,帮助学生掌握体内药物与毒物分析基本技能,提高教学效果.

目的:基于柱色谱技术和液相色谱-离子阱-飞行时间质谱(LCMS-IT-TOF)技术展开地榆的物质基础研究,分析地榆水提液中不同成分在D101型大孔树脂和聚酰胺树脂上的分布规律.方法:通过D101型大孔树脂和聚酰胺树脂分离地榆水提液,使用LCMS-IT-TOF检测,使用ACQUITY UPLC HSS T3色谱柱(2.1 mm×100 mm,1.8 μm),流动相水-乙腈梯度洗脱,流速0.3 mL· min-1,柱温30℃,采用电喷雾离子源(ESI),正、负离子模式采集数据,质谱扫描范围m/z 100～1 200,根据精确相对分子质量及质谱碎片等信息并结合文献研究,鉴定地榆水提液的上样流出液和洗脱液中化学成分.提取各样品中质谱峰强度数据,绘制成分在各流分中分布热图,直观比较各成分的洗脱规律.结果:D101型大孔树脂和聚酰胺树脂的富集和分离作用明显,鞣质类成分主要集中于大孔树脂上样流出液及其水洗脱液中,三萜类成分主要分布在大孔树脂90％乙醇洗脱液中.在地榆水提液的流出液和洗脱液中,共鉴定出63个化合物,其中6-O-galloylnorbergerin,3-O-galloylnorbergerin,2,6-乙酰氧基-5,7-二羟基-8-甲氧基色原酮,鞣花酸-4-吡喃阿拉伯糖苷或其异构体,2-甲基-3-乙酰氧基-苯并呋喃-5,6-O-硫酸酯6个成分在地榆中为首次发现.结论:该方法能够快速准确地鉴定地榆水提液经过柱色谱分离后的成分分布,为探索地榆药效成分及其作用机制提供了实验依据.

Effective treatment of brain metastases is hindered by the blood-brain barrier (BBB) and the rapid development of resistance to drug therapy. Moreover, the clinical application of general formulations is hampered by biological barriers and biological elimination. To tackle this challenge, we report a feasible approach for the assembly of polymer-covalent organic framework (COF) nanocomposites into 150 nm thin platelets as a drug delivery vehicle for enhanced retention in brain tumours. Using intravital imaging, we demonstrate that these polymer-COF nanocomposites are able to traverse the BBB in mice and achieve direct tumour accumulation in intracranial orthotopic models of brain metastasis from renal cancer (BMRC). These nanocomposites can target brain tumour cells and respond to tumour microenvironmental characteristics, including acidic and redox conditions. Intracranial tumour acidity triggers the breakdown of the nanoassemblies to polymer-COF nanocomposites due to the presence of borate bonds. Furthermore, in vivo studies on the nanocomposites showed enhanced brain tumour-targeting efficiency and therapeutic effects compared to those of free-drug dosing. Mice treated with drug-loaded polymer-COF nanocomposites also show protection from systemic drug toxicity and improved survival, demonstrating the preclinical potential of this nanoscale platform to deliver novel combination therapies to BMRC and other central nervous system (CNS) tumours.

目的 考察罗汉果苷V作为新型载体对难溶性药物的增溶作用.方法 以罗汉果苷V为载体,紫杉醇为模型药物,采用溶剂法制备紫杉醇-罗汉果苷V固体分散体.采用高效液相色谱(HPLC)法测定紫杉醇的含量,考察固体分散体中紫杉醇的饱和溶解度和体外溶出性能的变化.同时,用差示热扫描法(DSC)进行物相鉴别,评价药物在固体分散体中晶型变化.结果 紫杉醇-罗汉果苷Ⅴ固体分散体的饱和溶解度比紫杉醇增加约375倍;与紫杉醇比较,固体分散体的体外溶出速率和累积溶出度明显提高;差示热扫描法结果表明,紫杉醇在固体分散体内以无定形存在.结论 罗汉果苷V能明显增加难溶性药物的溶解度和体外溶出度,且该载体安全、无毒,有望成为难溶性药物增溶的新型载体.

OBJECTIVES:Reproducing human hair follicles in vitro is often limited by various reasons such as the lack of a systematic approach to culture distinct hair follicle cell types to reproduce their spatial relationship. Here, we reproduce hair follicle-like constructs resembling the spatial orientation of different cells in vivo, to study the role of keratinocytes in maintaining cellular compartmentalization among hair follicle-related cells.MATERIALS AND METHODS:Dermal papilla (DP) cells, HaCaT keratinocytes and human dermal fibroblast (HDF) cells were seeded sequentially into three-dimensional (3D) microwells fabricated from polyethylene glycol diacrylate hydrogels. Quantitative polymerase chain reaction was used to compare inductive gene expression of 3D and two-dimensional (2D) DP. DP and HaCaT cells were transfected with green fluorescent protein and red fluorescent protein lentivirus, respectively, to enable cell visualization using confocal microscopy.RESULTS:The 3D DP cultures showed significantly enhanced expression of essential DP genes as compared 2D cultures. Core-shell configurations containing keratinocytes forming the outer shell and DP forming the core were observed. Migratory polarization was mediated by cell-cell interaction between the keratinocytes and HDF cells, while preserving the aggregated state of the DP cells.CONCLUSIONS:Keratinocytes may play a role in maintaining compartmentalization between the DP and the surrounding HDF residing in the dermis, and therefore maintains the aggregative state of the DP cells, necessary for hair follicle development and function.

A matrix metalloproteinases (MMPs) triggered multifunctional micellar system loading traditional chemotherapeutic agent paclitaxel (PTX) and marimastat (MATT), an inhibition of matrix metalloproteinases, was established to prevent tumor growth and metastasis. The micelles were self-assembled by the conjugate synthesized with an MMPs sensitive peptide as the bridge between PTX and poly(ethylene glycol) (PEG). 4T1 cell line derived from a murine breast tumor was selected as the cell model due to its high metastasis. The cytotoxicity assay and cell apoptosis analysis revealed that the sensitive micelles increased the toxicity and cell apoptosis compared with the insensitive micelles. In addition, this system also exhibited high penetration ability in tumor spheroids and significant invasion inhibition with the method of Transwell invasion assay. In the BALB/c mice bearing 4T1 tumors, this system inhibited the growth of the metastatic tumor and prevented the incidence of lung metastasis with low systemic toxicity. And the expression of MMP-2 and MMP-9, which are important in the process of tumor metastasis, was down-regulated. Generally, the data obtained from the in vitro and in vivo studies confirmed that the codelivery of PTX and MATT by the MMPs sensitive micelles achieved not only significant antitumor effect but also obvious inhibition effect of tumor metastasis, which are the leading cause of cancer deaths. This functional particle system may provide a promising strategy for metastatic breast cancer therapy.

Transdermal drug delivery is widely investigated as an alternative drug administration route to oral delivery and hypodermic injections. Owing to the availability of human skin samples, in vitro tests are used to predict the in vivo delivery of transdermal drugs. The most widely used validation method is skin permeation using diffusion cells. Traditional diffusion cells, however, are capacious and often require large amounts of skin sample and drugs, which is undesirable, given the scarcity of new drug entities and the limitation of skin sample supply. In this study, we fabricated miniaturized multichannel devices (MCDs) by 3D printing, to minimize the use of skin and drug samples. The MCDs were compared with conventional static diffusion cells and achieved comparable drug permeation profiles. The finite element method-based simulation revealed the efficient carry-off of permeated ingredients by the multichannel devices, and a critical role of distance between the buffer stream and skin sample in determining the flow velocity inside the chamber. The results support these devices as qualified alternatives to Franz cells for in vitro permeation studies using biomembranes, with reduced use of skin and drug samples.

目的 探讨附子主要毒性生物碱成分乌头碱、新乌头碱和次乌头碱的肝脏摄取转运机制.方法 以HepG2细胞为体外模型,进行细胞摄取研究,并用液相色谱-质谱串联(LC-MS/MS)法测定细胞内的药物浓度.结果HepG2细胞对乌头碱、新乌头碱和次乌头碱的摄取具有温度、浓度和pH依赖性,化学解耦联剂FCCP和能量抑制剂叠氮化钠可显著抑制乌头碱、新乌头碱和次乌头碱的细胞摄取.有机阳离子药物如奎尼丁、维拉帕米、吡拉明、苯海拉明及金刚烷胺可显著抑制3种生物碱的摄取,但有机阳离子转运蛋白(OCTs)、有机阳离子/肉毒碱转运蛋白(OCTNs)、多药及毒素外排转运蛋白(MATEs)和质膜单胺转运蛋白(PMAT)的典型底物或抑制剂对附子生物碱的细胞摄取没有抑制作用.结论 附子毒性生物碱进入肝细胞以主动转运为主,新型有机阳离子/质子逆向转运蛋白可能是其进入肝脏的分子基础.

目的 建立测定去水卫矛醇含量的柱前衍生化反相高效液相色谱法(RP-HPLC).方法 去水卫矛醇经二乙基二硫代氨基甲酸钠衍生化后进行RP-HPLC分析,采用Welch Ultimate XB-CN色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈-水(50:50),流速为1.0 mL?min-1,检测波长为278 nm.结果 40℃下衍生化反应可在30 min内完成,去水卫矛醇衍生物在10 h内稳定.去水卫矛醇在0.100～20.0μg?mL-1范围内线性关系良好(r=0.9998).检测限(LOD)和定量限(LOQ)分别为30和100 ng?mL-1.方法专属性、准确度和精密度均符合要求.结论 该方法快速、准确、灵敏,具有良好的专属性和重复性,可用于去水卫矛醇的质量控制.

Sleep electroencephalography (EEG) provides an opportunity to study sleep scientifically, whose chaotic, dynamic, complex, and dissipative nature implies that non-linear approaches could uncover some mechanism of sleep. Based on well-established complexity theories, one hypothesis in sleep medicine is that lower complexity of brain waves at pre-sleep state can facilitate sleep initiation and further improve sleep quality. However, this has never been studied with solid data. In this study, EEG collected from healthy subjects was used to investigate the association between pre-sleep EEG complexity and sleep quality. Multiscale entropy analysis (MSE) was applied to pre-sleep EEG signals recorded immediately after light-off (while subjects were awake) for measuring the complexities of brain dynamics by a proposed index, CI1−30. Slow wave activity (SWA) in sleep, which is commonly used as an indicator of sleep depth or sleep intensity, was quantified based on two methods, traditional Fast Fourier transform (FFT) and ensemble empirical mode decomposition (EEMD). The associations between wake EEG complexity, sleep latency, and SWA in sleep were evaluated. Our results demonstrated that lower complexity before sleep onset is associated with decreased sleep latency, indicating a potential facilitating role of reduced pre-sleep complexity in the wake-sleep transition. In addition, the proposed EEMD-based method revealed an association between wake complexity and quantified SWA in the beginning of sleep (90 min after sleep onset). Complexity metric could thus be considered as a potential indicator for sleep interventions, and further studies are encouraged to examine the application of EEG complexity before sleep onset in populations with difficulty in sleep initiation. Further studies may also examine the mechanisms of the causal relationships between pre-sleep brain complexity and SWA, or conduct comparisons between normal and pathological conditions.