Journal of pharmaceutical and biomedical analysis最新文献_第9页

Development and validation of an UPLC-MS/MS method with polarity switching for simultaneous determination of 14 antiepileptic drugs and 2 metabolites in human serum 同时测定人血清中14种抗癫痫药物和2种代谢物的极性切换UPLC-MS/MS方法的建立与验证

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-02 DOI: 10.1016/j.jpba.2024.116655

Xiao-han Peng , Yan-lin Zhao , Zhong Huang , Xin-feng Xia , Kun Wang , Peng Jin , Yan Du , Dao-quan Tang

Currently, treatment with antiepileptic drugs (AEDs) is still the first choice for epileptic patients, while monitoring their blood concentrations is undoubtedly beneficial for minimizing their adverse side effects and optimizing their therapeutic effects. In this study, an ultra-high performance liquid chromatography coupled with tandem mass spectrometry with polarity switching was developed and validated for simultaneous determination of 14 AEDs and 2 active metabolites in human serum. Olanzapine was selected as the internal standard. One-step protein precipitation using methanol containing 0.05 % formic acid was used to treat sample, and the supernatant was injected for analysis without further evaporation and reconstitution. Chromatographic separation was performed on an Aglient Zorbax Eclipse Plus C18 (50 mm × 2.1 mm, 1.8 μm) column with gradient methanol and 0.1 % formic acid in water as mobile phase. Multi-reaction monitoring was performed for quantification of 16 analytes in polarity switching mode. Matrix-matched calibration curves of 16 analytes presented good linearity within the test concentration range (r > 0.99). The intra- and inter-run accuracies and precisions at the lower limit of quantification, and low, medium and high quality control levels were all less than 20 % or 15 %, respectively. The extraction recovery, matrix effect, and stability were all acceptable under detected conditions. Finally, this method was successfully applied in the quantitation of target analytes in the serum of patients received AEDs.

目前，抗癫痫药物治疗仍是癫痫患者的首选，而监测抗癫痫药物的血药浓度无疑有助于减少其不良副作用，优化其治疗效果。本研究建立了一种极性切换的超高效液相色谱-串联质谱联用方法，用于同时测定人血清中14种AEDs和2种活性代谢物。选择奥氮平作为内标。样品采用含0.05 %甲酸的甲醇一步蛋白沉淀法处理，上清液无需进一步蒸发和重构即可进样分析。色谱柱为Aglient Zorbax Eclipse Plus C18(50 mm × 2.1 mm, 1.8 μm)，流动相为梯度甲醇和0.1 %甲酸。在极性切换模式下对16种分析物进行多反应监测。16种分析物的基质匹配校准曲线在检测浓度范围内呈良好的线性关系（r > 0.99）。定量下限、低、中、高质量控制水平的批内、批间准确度和精密度分别小于20 %和15 %。在检测条件下，提取回收率、基质效应和稳定性均可接受。最后，该方法成功应用于aed患者血清中目标分析物的定量。

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引用次数: 0

Acquisition and transcriptomic analysis of tissue micro-regions using a capillary-based method 使用基于毛细管的方法获取和组织微区转录组学分析。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2024-12-31 DOI: 10.1016/j.jpba.2024.116656

Kaiqiang Ye , Yunxia Guo , Ying Wang , Jitao Xu , Qingyang Qin , Liyong He , Xi Yang , Yan Huang , Qinyu Ge , Xiangwei Zhao

Profiling the site-specific transcriptomes of microregions of interest (mROIs) contributes to a complete understanding of multicellular organisms. However, the simple and efficient isolation of mROIs for spatially detecting gene expression remains challenging. Here, we develop an efficient capillary-based microdissection system (CMS) for precisely isolating targeted samples from tissue sections. Optimized sampling procedures reveal that CMS can perform mROI isolation with an efficiency of 97.9 %, and detect a sufficient number of genes for gene expression profiling (CMS-seq). We apply CMS-seq to uncover spatial heterogeneity in the cortex region of the mouse, and the subregions of hippocampus in an Alzheimer's disease (AD) mouse. Results demonstrate that CMS-seq can profile spatial transcriptomes in tissue sections and holds promise for application spatial multi-omics.

分析感兴趣微区（mris）的位点特异性转录组有助于对多细胞生物的全面了解。然而，简单有效地分离核磁共振成像用于空间检测基因表达仍然是一个挑战。在这里，我们开发了一种高效的基于毛细血管的显微解剖系统（CMS），用于从组织切片中精确分离目标样本。优化后的采样程序表明，CMS可以以97.9% %的效率进行mROI分离，并检测到足够数量的基因用于基因表达谱（CMS-seq）。我们应用CMS-seq揭示了阿尔茨海默病（AD）小鼠皮质区域和海马亚区的空间异质性。结果表明，CMS-seq可以在组织切片中分析空间转录组，并有望应用于空间多组学。

引用次数: 0

Determination of carbamazepine profile in human plasma by GC-MS 气相色谱-质谱法测定人血浆中卡马西平的含量。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2024-12-31 DOI: 10.1016/j.jpba.2024.116658

Zeynep Özdemir, Selen Al, Aykut Kul , Olcay Sagirli

Epilepsy is a major disease affecting millions of people worldwide. Carbamazepine is on the World Health Organization's list of essential medicines and is one of the most prescribed medicines for treating epilepsy. It has a narrow therapeutic range (4–12 μg/mL). Due to this narrow therapeutic range, toxic and adverse reactions are likely to be observed in the clinic. Therefore, therapeutic drug monitoring (TDM) should be routinely performed in the clinic for epilepsy patients treated with carbamazepine. Considering that an antiepileptic drug produces an antiepileptic effect only when its free (non-protein bound) concentration crosses the blood-brain barrier and reaches the brain, knowing and measuring the free drug fraction is important. In this study, a GC-MS method was developed for TDM of total and free carbamazepine, and carbamazepine epoxide in plasma. Free carbamazepine and carbamazepine epoxide were collected by ultra-filtrate, and analytes were extracted in plasma using salt-assisted liquid-liquid extraction (SALLME). The method was validated according to the European Medicines Agency (EMA) bioanalytical method validation guidelines. In the developed method, calibration curves were constructed for total carbamazepine, free carbamazepine, total carbamazepine epoxide, and free carbamazepine epoxide with calibration ranges of 1–20 µg/mL, 0.25–20 µg/mL, 0.4–8 µg/mL, and 0.1–8 µg/mL, respectively. The corresponding LLOQ values were 1, 0.25, 0.4, and 0.1 µg/mL. The correlation coefficient for both molecules was > 0.99 and the developing technique was applied to TDM for carbamazepine profile for plasma of patient samples.

癫痫是影响全世界数百万人的一种主要疾病。卡马西平被列入世界卫生组织的基本药物清单，是治疗癫痫最常用的处方药之一。治疗范围窄（4 ~ 12 μg/mL）。由于这种狭窄的治疗范围，在临床中很可能观察到毒性和不良反应。因此，临床对卡马西平治疗的癫痫患者应常规进行治疗性药物监测（TDM）。考虑到抗癫痫药物只有在其游离（非蛋白结合）浓度穿过血脑屏障并到达大脑时才产生抗癫痫作用，了解和测量游离药物部分是很重要的。本研究建立了血浆中总卡马西平、游离卡马西平和环氧卡马西平的TDM测定方法。用超滤液收集游离卡马西平和环氧卡马西平，用盐辅助液-液萃取法（salme）在血浆中提取分析物。该方法根据欧洲药品管理局（EMA）生物分析方法验证指南进行验证。建立了总卡马西平、游离卡马西平、总卡马西平环氧化物和游离卡马西平环氧化物的标定曲线，标定范围分别为1 ~ 20 µg/mL、0.25 ~ 20 µg/mL、0.4 ~ 8 µg/mL和0.1 ~ 8 µg/mL。相应的定量限分别为1、0.25、0.4、0.1 µg/mL。两种分子的相关系数为> 0.99，并将开发技术应用于患者血浆卡马西平谱的TDM。

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引用次数: 0

A simple automated assay format for measuring multiple immune checkpoint inhibitors 用于测量多种免疫检查点抑制剂的简单自动分析格式。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2024-12-30 DOI: 10.1016/j.jpba.2024.116657

Ragnhild V. Nome , Øystein Flatebø , Sigurd Leinæs Bøe , Rolf Anton Klaasen , Elin Aamdal , Marius Normann , Nils Bolstad , David John Warren

Immune checkpoint inhibitors (ICIs) have improved survival rates in oncology, but there is a rising concern for immune-related adverse health outcomes. Monitoring drug serum concentration would enable tailored dosing, however this strategy has not yet been evaluated for ICIs. Fully automated analyte capture assays with time-resolved fluorometry using protein A as tracer, were developed for three different ICIs; the cytotoxic T lymphocyte Antigen-4 (CTLA4) inhibitor ipilimumab (Yervoy; Bristol-Myers Squibb) and the Programmed Death-1 (PD-1) inhibitors nivolumab (Opdivo; Bristol-Myers Squibb) and pembrolizumab (Keytruda; Merck). Drug trough levels were measured in serum samples from ICI-treated patients. Measuring ranges were 1–100 mg/L for all three drugs. Automation allowed for 110 samples to be analyzed in < 4 h. Median drug trough-levels after 5–7 weeks of treatment were 20 (range <1.0–45) mg/L for ipilimumab (n = 113), 60 (range 14–75) mg/L) for nivolumab (n = 21) and 19 (range 7.4–39) mg/L for pembrolizumab (n = 20). Routine drug concentration monitoring for ipilimumab, nivolumab and pembrolizumab is feasible using fully automated analyte capture assays constructed with commercially available reagents. The large drug serum concentration ranges in samples from real-world patients, should be further investigated to assess the clinical relevance of ICI concentration monitoring.

免疫检查点抑制剂（ICIs）提高了肿瘤患者的生存率，但对免疫相关的不良健康结果的担忧日益增加。监测药物血清浓度将使量身定制的剂量，然而，这一策略尚未被评估为ici。全自动分析物捕获测定与时间分辨荧光法使用蛋白A作为示踪剂，开发了三种不同的ici；细胞毒性T淋巴细胞抗原4 （CTLA4）抑制剂伊匹单抗(Yervoy；Bristol-Myers Squibb)和程序性死亡-1 （PD-1）抑制剂nivolumab (Opdivo；百时美施贵宝（Bristol-Myers Squibb）和pembrolizumab (Keytruda；默克公司)。在ci治疗患者的血清样本中测量药物谷水平。三种药物的测量范围均为1 ~ 100 mg/L。自动化允许对110个样品进行分析

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引用次数: 0

A simple LC-MS/MS assay for the quantification of E6011, a novel anti-fractalkine monoclonal antibody, in cynomolgus monkey serum - comparison with ligand binding assay LC-MS/MS法定量食蟹猴血清中一种新型抗fractalkine单克隆抗体E6011——与配体结合法的比较

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2024-12-30 DOI: 10.1016/j.jpba.2024.116659

Kenji Kita , Haruna Ono , Tomoko Kojima , Yuji Mano

E6011 is a monoclonal antibody that is currently under development for the treatment of rheumatoid arthritis. While ligand binding assays (LBAs) are typically employed for the determination of therapeutic antibodies, ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) represents an alternative platform. E6011 in monkey serum was treated with ammonium sulfate to obtain pellets for subsequent processing. The pellets were subjected to denaturalization reduction, alkylation, and tryptic digestion. The resulting signature peptide of E6011, TLADGVPSR, was assayed. The pellet digestion assay was validated in accordance with the established bioanalytical guidelines. E6011 in monkey serum was quantifiable from 3 to 729 µg/mL, with a sample volume of 0.02 mL. The selectivity was confirmed in 12 individual monkey sera. The accuracy and precision were within ± 11.2 % and 15.0 %, respectively. The validated UPLC-MS/MS assay was employed in a pharmacokinetic study in monkeys. After the intravenous dose at 1 mg/kg, E6011 reached the maximum of 27.4 μg/mL, then declined with the half-life of 169 h. The serum E6011 concentrations determined by the UPLC-MS/MS were comparable to those obtained by the LBA with electrochemiluminescence detection. These findings suggest that the established simple UPLC-MS/MS assay is reproducible and can serve as an alternative assay platform.

E6011是一种单克隆抗体，目前正在开发中，用于治疗类风湿性关节炎。虽然配体结合试验（LBAs）通常用于测定治疗性抗体，超高效液相色谱串联质谱（UPLC-MS/MS）代表了另一种平台。用硫酸铵处理猴子血清中的E6011，得到后续加工用的微球。球团进行变性还原、烷基化和胰蛋白酶消化。检测了E6011的特征肽TLADGVPSR。根据建立的生物分析指南验证了颗粒消化试验。猴子血清中E6011的定量范围为3 ~ 729 µg/mL，样品体积为0.02 mL。在12个单独的猴血清中证实了选择性。准确度和精密度分别在± 11.2 %和15.0 %以内。采用经验证的UPLC-MS/MS法对猴子进行了药代动力学研究。静脉给药1 mg/kg后，E6011达到最大值27.4 μg/mL，随后逐渐下降，半衰期为169 h。UPLC-MS/MS测定的血清E6011浓度与电化学发光法测定的血清E6011浓度相当。这些结果表明，所建立的简单UPLC-MS/MS分析具有可重复性，可以作为替代的分析平台。

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引用次数: 0

Validation of a liquid chromatography-high-resolution mass spectrometry method to quantify peptide-related impurities in teriparatide 液相色谱-高分辨率质谱法定量特立帕肽中肽相关杂质的验证。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2024-12-28 DOI: 10.1016/j.jpba.2024.116654

Daniel A. Weisz , Sarah M. Rogstad , Kui Zeng , Eric Pang , Ilan Geerlof-Vidavsky

With recent advances in quantitative high-resolution mass spectrometry (HRMS), there is growing interest in developing liquid chromatography (LC)-HRMS methods that can simultaneously quantify numerous critical impurities in a peptide or protein drug. This approach is attractive as it could reduce the total number of methods and instruments required during product development and quality control testing, while taking advantage of the technique’s high specificity and sensitivity. To investigate the feasibility of this approach for peptide drugs, an LC-HRMS method was validated for the quantification of six peptide-related impurities in teriparatide, the 34-amino acid active ingredient in Forteo. External calibration curves were constructed to correlate the peak area ratio of impurity-to-teriparatide to a known impurity abundance. The method displayed good specificity, sensitivity, linearity, accuracy, repeatability, intermediate precision, and robustness. The lower limits of quantification were 0.02 % or 0.03 % of teriparatide, below the regulatory reporting threshold of 0.10 %. It was found that quantification using three isotopic peaks per peptide did not provide a significant benefit over quantification with one isotopic peak. The method was validated successfully without the impractical inclusion of an isotopically-labeled internal standard for each impurity. Future studies will be conducted to determine the method’s longer-term reproducibility.

随着定量高分辨率质谱法（HRMS）的最新进展，人们对开发液相色谱(LC)-HRMS方法越来越感兴趣，这种方法可以同时定量多肽或蛋白质药物中的许多关键杂质。这种方法很有吸引力，因为它可以减少产品开发和质量控制测试过程中所需的方法和仪器的总数，同时利用该技术的高特异性和灵敏度。为了考察该方法在多肽类药物中的可行性，采用LC-HRMS方法对三立帕肽（Forteo中含有34个氨基酸的活性成分）中6种多肽相关杂质进行了定量验证。构建外部校准曲线，将杂质-特立帕肽的峰面积比与已知杂质丰度相关联。该方法具有良好的特异度、灵敏度、线性度、准确度、重复性、中间精密度和鲁棒性。定量下限为特立帕肽的0.02 %或0.03 %，低于监管报告阈值0.10 %。结果发现，每个肽使用三个同位素峰的定量并不比使用一个同位素峰的定量提供显著的好处。该方法被成功地验证了，没有为每个杂质不切实际地包含同位素标记的内标。未来的研究将确定该方法的长期可重复性。

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引用次数: 0

Untargeted metabolomics combined with lipidomics revealed the effects of myocardial infarction and exercise rehabilitation on blood circulation metabolism of patients based on liquid chromatography-mass spectrometry 非靶向代谢组学联合脂质组学基于液相色谱-质谱分析揭示心肌梗死和运动康复对患者血液循环代谢的影响。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2024-12-26 DOI: 10.1016/j.jpba.2024.116651

Na Wang , Huimin Li , Hao Wu , Zilin Xia , Dabing Ren , Yunmei Zhang , Yan Zhao , Hong Zhang , Ke Zhuang , Lunzhao Yi

Myocardial infarction (MI) is a major cause of death worldwide. Exercise rehabilitation (ER) is a powerful tool to improve life quality and prognosis of MI patients. Herein, we developed an untargeted metabolomics combined with lipidomics method to qualitatively and quantitatively detect metabolites in plasma. A total of 475 metabolites were annotated according to MS, MS/MS, and quantified by internal standard method. Moreover, medical statistical methods combined with chemometrics were used for metabolomics data mining and interpretation of clinical issues (matched Cohort 1, n = 90, Cohort 2, n = 6). The results illustrated that abnormal lipid metabolism is the most significant metabolic disorder for MI patients. And, three metabolic pathways, bile secretion, HIF-1 signaling pathway, and glutathione metabolism were uncovered in MI patients. Furthermore, glutamine, Phenylacetylglutamine (PAGln) and lysophosphatidylcholine (LPCs) were revealed as the essential biomarkers for ER of MI patients. Our findings revealed the metabolic landscape of MI and metabolic alterations after ER, will underlay potential applications of plasma metabolites in the detection of MI and optimization of ER program.

心肌梗死（MI）是世界范围内的一个主要死亡原因。运动康复是改善心梗患者生活质量和预后的有力手段。在此，我们开发了一种非靶向代谢组学结合脂质组学的方法来定性和定量检测血浆中的代谢物。采用MS、MS/MS对475种代谢物进行注释，内标法定量。此外，使用医学统计学方法结合化学计量学进行代谢组学数据挖掘和临床问题解释（匹配队列1，n = 90，队列2，n = 6）。结果表明，脂质代谢异常是心肌梗死患者最显著的代谢紊乱。发现了MI患者的胆汁分泌、HIF-1信号通路和谷胱甘肽代谢3条代谢途径。此外，谷氨酰胺、苯乙酰谷氨酰胺（PAGln）和溶血磷脂酰胆碱（LPCs）是心肌梗死患者ER的重要生物标志物。我们的研究结果揭示了心肌梗死的代谢景观和ER后的代谢改变，将为血浆代谢物在心肌梗死检测和ER程序优化中的潜在应用奠定基础。

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引用次数: 0

Analysis of Pz-1, a promising therapeutic for organophosphorus poisoning from rodent plasma by liquid chromatography-tandem mass spectrometry 有前途的治疗鼠类血浆有机磷中毒药物Pz-1的液相色谱-串联质谱分析。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2024-12-26 DOI: 10.1016/j.jpba.2024.116650

Khadija Bilkis , Moustafa M.R. Khalaf , Darci M. Fink , Jeremy W. Chambers , Brian A. Logue

Organophosphorus (OP) pesticides (e.g., parathion) and nerve agents (e.g., soman) can produce acute and long-term neurological problems. Exposure to OP chemicals is responsible for an estimated 200,000 deaths annually. Pz-1 (N-(5-(tert butyl)isoxazol-3-yl)-2-(4-(5-(1-methyl-1H-pyrazol-4-yl)-1H-benzo[d]imidazol-1-yl)phenyl)acetamide) is a muscle specific kinase (MuSK) inhibitor which has shown potential as a treatment for OP chemical exposure and as a tyrosine kinase inhibitor to impede the growth of cancer cells. While development of this treatment requires the availability of a validated analytical method, no method currently exists for analysis of Pz-1 from biological samples. In this study, an analytical method was developed for Pz-1 from rat (and mouse) plasma. Plasma was prepared by precipitating plasma proteins, isolating the supernatant, evaporating to dryness and reconstituting in 1:1 MeOH:water. Prepared samples were analyzed by reversed-phase liquid chromatography tandem mass-spectrometry (LC-MS/MS). The method produced excellent sensitivity, with a limit of detection of 1 nM (455 ng/L). The calibration range was 3–100 nM and the calibration curve produced excellent linear behavior (R² ≥ 0.99 and PRA ≥ 91 %). The method also showed good accuracy and precision. The validated method was used to detect Pz-1 in mouse plasma following intraperitoneal (IP) treatment with 5 mg/kg Pz-1. In summary, this method shows promise as a simple and sensitive method to analyze Pz-1 in rat plasma to facilitate its continued development as a treatment for OP toxicity.

有机磷（OP）杀虫剂（如对硫磷）和神经毒剂（如索曼）可产生急性和长期的神经问题。据估计，每年因接触 OP 化学品而死亡的人数达 20 万。Pz-1（N-(5-(叔丁基)异恶唑-3-基)-2-(4-(5-(1-甲基-1H-吡唑-4-基)-1H-苯并[d]咪唑-1-基)苯基)乙酰胺）是一种肌肉特异性激酶（MuSK）抑制剂，已显示出治疗 OP 化学品暴露和作为酪氨酸激酶抑制剂阻碍癌细胞生长的潜力。这种治疗方法的开发需要有效的分析方法，但目前还没有从生物样本中分析 Pz-1 的方法。本研究开发了一种从大鼠（和小鼠）血浆中检测 Pz-1 的分析方法。血浆的制备方法是沉淀血浆蛋白，分离上清液，蒸发至干，用 1:1 MeOH:water 复溶。制备好的样品采用反相液相色谱串联质谱法（LC-MS/MS）进行分析。该方法灵敏度极高，检测限为 1 nM（455 ng/L）。校准范围为 3-100 nM，校准曲线线性良好（R2 ≥ 0.99，PRA ≥ 91 %）。该方法还具有良好的准确度和精密度。在小鼠腹腔注射（IP）5 mg/kg Pz-1 后，采用该方法检测了小鼠血浆中的 Pz-1。总之，该方法是分析大鼠血浆中 Pz-1 的一种简单而灵敏的方法，有助于继续开发其作为 OP 毒性的治疗方法。

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引用次数: 0

An exploratory multi-omics study reveals distinct molecular signatures of ulcerative colitis and Crohn's disease and their correlation with disease activity 一项探索性多组学研究揭示了溃疡性结肠炎和克罗恩病的不同分子特征及其与疾病活动的相关性。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2024-12-25 DOI: 10.1016/j.jpba.2024.116652

Nguyen Tran Nam Tien , Eun Jeong Choi , Nguyen Quang Thu , Seung Jung Yu , Duc Ninh Nguyen , Dong Hyun Kim , Nguyen Phuoc Long , Hong Sub Lee

Clinically heterogeneous spectrum and molecular phenotypes of inflammatory bowel disease (IBD) remain to be comprehensively elucidated. This exploratory multi-omics study investigated the serum molecular profiles of Crohn's disease (CD) and ulcerative colitis (UC), in association with elevated fecal calprotectin and disease activity states. The serum proteome, metabolome, and lipidome of 75 treated IBD patients were profiled. Single- and multi-omic data analysis was performed to determine differential analytes and integrative biosignatures for biological interpretations. We found that chronic inflammation, phosphatidylcholines and bile acid homeostasis disturbances underlined the differences between CD and UC. Besides, elevated calprotectin was associated with higher levels of inflammatory proteins and sphingomyelins (SM) and lower levels of bile acids, amino acids, and triacylglycerols (TG). Relative to the remission disease state, the active form was characterized by decreased abundances of SMs and increased abundances of inflammatory proteins and TGs. We also observed that molecular changes upon treatment escalation were putatively related to altered levels of inflammatory response proteins, amino acids, and TGs. ISM1, ANGPTL4, chenodeoxycholate, Cer(18:1;2 O/24:1), and TG were identified as candidates subject to further investigation. Altogether, our study revealed that disturbances in immune response, bile acid homeostasis, amino acids, and lipids potentially underlie the clinically heterogeneous spectrum of IBD.

炎症性肠病（IBD）的临床异质性谱和分子表型仍有待全面阐明。这项探索性多组学研究调查了克罗恩病（CD）和溃疡性结肠炎（UC）的血清分子谱，它们与粪便钙保护蛋白升高和疾病活动状态有关。对75例接受治疗的IBD患者的血清蛋白质组、代谢组和脂质组进行了分析。进行单组学和多组学数据分析以确定差异分析物和生物解释的综合生物特征。我们发现慢性炎症、磷脂酰胆碱和胆汁酸稳态紊乱突出了CD和UC之间的差异。此外，钙保护蛋白升高与炎症蛋白和鞘磷脂（SM）水平升高以及胆汁酸、氨基酸和甘油三酯（TG）水平降低有关。相对于疾病缓解状态，活性形式的特征是SMs丰度降低，炎症蛋白和tg丰度增加。我们还观察到，治疗升级后的分子变化可能与炎症反应蛋白、氨基酸和tg水平的改变有关。ISM1、ANGPTL4、鹅脱氧胆酸盐、Cer（18:1;2 O/24:1）和TG被确定为有待进一步研究的候选。总之，我们的研究揭示了免疫反应、胆汁酸稳态、氨基酸和脂质的紊乱可能是IBD临床异质性谱的基础。

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引用次数: 0

Metabolic and molecular basis of flavonoid biosynthesis in Lycii fructus: An integration of metabolomic and transcriptomic analysis 枸杞类黄酮生物合成的代谢和分子基础：代谢组学和转录组学的综合分析。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2024-12-24 DOI: 10.1016/j.jpba.2024.116653

Limei Tong , Yinxiu Jiang , Xinrun Zhang , Xia Zhang , Wenhua Zhang , Gang Ren , Zhanping Chen , Yuling Zhao , Sheng Guo , Hui Yan , Yang Pan , Jin-ao Duan , Fang Zhang

Flavonoids serve as bioactive components and contribute to medicinal and nutritional profile of Lycii fructus. However, there is limited information regarding the influence of ecological environments on the flavonoid biosynthesis pathway. In this study, we integrated transcriptome sequencing and metabonomic techniques across three distinct cultivation regions to elucidate the processes of flavonoids biosynthesis and the associated gene expression levels in L. fructus. LC-MS/MS based metabolomics revealed significant variations in metabolite profiles including 43 differential flavonoid metabolites, predominantly consisting of flavanol compounds across diverse regions. Additionally, 154 significantly differentially expressed genes (DEGs) were categorized in the flavonoid biosynthesis identified by de novo transcriptome assembly. Transcription factors C2C2 MYB, NAC, WRKY, AP2/ERF and B3 superfamily were the mainly hub genes regulating the flavonoids biosynthesis. The flavonoid pathway was built through integrated analysis of DEGs and DAMs to illustrate the molecular mechanism of flavonoid biosynthesis. Precipitation and temperature may serve as the primary environmental factors that affected the flavonoids variations. This study proposed a schematic of flavonoid biosynthesis in L. fructus, and further provided evidence for environmental response of L. fructus.

黄酮类化合物是枸杞子的活性成分，具有丰富的药用价值和营养价值。然而，关于生态环境对类黄酮生物合成途径的影响的信息有限。在这项研究中，我们整合了三个不同栽培区域的转录组测序和代谢组学技术，以阐明L. fructus中黄酮类化合物的生物合成过程和相关基因的表达水平。基于LC-MS/MS的代谢组学发现，不同地区的代谢物谱存在显著差异，包括43种不同的类黄酮代谢物，主要由黄烷醇化合物组成。此外，通过从头转录组组装鉴定，154个显著差异表达基因（DEGs）在类黄酮生物合成中被分类。转录因子C2C2、MYB、NAC、WRKY、AP2/ERF和B3超家族是调控黄酮类化合物生物合成的主要枢纽基因。通过对DEGs和dam的综合分析，构建了类黄酮途径，阐明了类黄酮生物合成的分子机制。降水和温度可能是影响黄酮类化合物变化的主要环境因子。本研究提出了枸杞类黄酮的生物合成原理图，为枸杞的环境响应提供了进一步的依据。

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引用次数: 0