益母草中阿魏酸对含益母草中成药质量标准影响的探讨

目的：建立HPLC/TQ-MS测定益母草中阿魏酸含量的方法并测定不同产地益母草中阿魏酸含量，评价益母草中阿魏酸对中成药调经止痛片及新生化颗粒的质量影响，为制定含有益母草并以阿魏酸为定量指标的中成药质量标准提供参考。方法：采用薄层色谱法及UPLC-Q/TOF定性鉴别及分析益母草中阿魏酸，采用HPLC/TQ-MS定量分析10批不同产地益母草中阿魏酸含量。基于定量分析结果及处方用量探讨益母草中阿魏酸含量对其中成药质量标准的影响。结果：薄层鉴别及UPLC-Q/TOF定性分析结果显示，10批不同产地益母草中均含有阿魏酸。HPLC/TQ-MS定量分析结果显示，益母草中阿魏酸含量在0.004 2%~0.006 5%之间。结论：益母草与当归、川芎等含有阿魏酸的药材配伍且其用量较大时，益母草中阿魏酸含量会对其质量评价产生一定的影响。     

Pharmacokinetics and brain penetration study of chlorogenic acid in rats

1. The present study is designed to investigate the brain distribution and plasma pharmacokinetics profiles of chlorogenic acid (CGA) after intranasal administration in Charles–Foster rats to evaluate whether the CGA molecules are transported directly via the nose-to-brain path.

2. The CGA is administered intravenously (IV) and intranasally (IN) at the dose of 10?mg/kg. Further, its concentration in the plasma, cerebrospinal fluid (CSF) and the whole brain is analyzed by HPLC-UV method.

3. The study observes that CGA is rapidly absorbed in plasma with t_max of 1?min similar to IV route after IN administration. The peak plasma concentration and AUC_0–24 are higher by 3.5 and 4.0 times respectively in IV administration, compared to IN delivery that represents the significant less systemic exposure of CGA in IN route.

4. However, the concentration of CGA in the brain is 4, 6.5, 5.3, 5.2 and 4.5 times higher at 30, 60, 120, 240 and 360?min, respectively in IN administration compared to IV administration. The exposure of CGA in the brain after IN administration (AUC_{brain, IN}) was significantly greater (4 times) as compared to the exposure of CGA in the brain (AUC_{brain, IV}) after IV administration reflecting significant brain uptake of CGA through nasal route. Therefore, IN delivery of CGA can be a promising approach for the treatment of stroke and neurodegenerative disorders.     

5-Aminolevulinic acid-mediated photodynamic activity in patient-derived cholangiocarcinoma organoids

BackgroundAccurate diagnosis of the disease extension of cholangiocarcinoma (CCA) is often difficult in clinical practice. The diagnostic yield of conventional pre-operative imaging or endoscopic procedures is sometimes insufficient for the evaluation of longitudinal spreading of CCA. Here we investigated the usefulness of 5-aminolevulinic acid (5-ALA) for the pre- or intra-operative diagnosis of CCA, using patient-derived organoids.MethodsFour CCA- and two adjacent tissue-derived organoids were established. After 5-ALA treatment, we assessed their photodynamic activity using fluorescence microscopy.ResultsCCA organoids established from different patients showed diverse morphology in contrast to monolayer structures of non-tumor organoids, and had the ability to form subcutaneous tumors in immunodeficient mice. CCA organoids demonstrated remarkably high photodynamic activity based on higher accumulation of protoporphyrin IX as a metabolite of 5-ALA compared to non-tumor organoids (40–71% vs. < 4%, respectively). Importantly, cancer cell-specific high photodynamic activity distinguished the organoids originated from biliary stenotic lesions from those of non-stenotic lesions in a CCA patient. The high photodynamic activity did not depend on the expression profile of heme biosynthesis genes.ConclusionsDistinct 5-ALA-based photodynamic activity could have diagnostic potential for the discrimination of CCA from non-tumor tissues.     

Topically injected adrenocorticotropic hormone induces mechanical hypersensitivity on a full‐thickness cutaneous wound model in rats

Cutaneous wound pain causes physical and psychological stress for patients with wounds. Previous studies reported that stress induces hyperalgesia and deteriorates wound healing. However, the effect of the stress response such as in hypothalamic‐pituitary‐adrenal (HPA) axis on local wound area is unclear. We aimed to investigate the effects of a stress response on the mechanical withdrawal threshold in the local wound area and describe the identification of a wound pain exacerbation. We topically injected adrenocorticotropic hormone (ACTH) into the granulation tissue of full‐thickness cutaneous wound model rats on the fifth day postwounding and measured the mechanical withdrawal thresholds, cytochrome P450 2Bs levels and concentration of 5,6‐epoxyeicosatrienoic acid in wound exudate. We found that ACTH induced mechanical hypersensitivity at 4 and 6 hours after injection (P = .004 and .021, respectively), and increased gene expression of cytochrome P450 2B12 expression (P = .046). Concentration of 5,6‐EET in the wound exudate was moderately correlated with the mechanical withdrawal threshold (r = ?.630). Finally, the mechanical withdrawal threshold in the 5,6‐EET group was significantly lower than that in the control group at 2 hours after the injection (P = .015). We propose that 5,6‐EET is one of the most promising contributors to the wound pain exacerbation. These findings could guide clinical wound and pain management.     

阿魏酸对创伤性脑损伤小鼠的神经保护作用

目的本研究利用创伤性脑损伤(traumatic brain injury,TBI)小鼠模型观察阿魏酸(FA)对TBI的神经保护作用,为防治TBI继发性损伤的药物开发提供新思路。方法利用C57/BL6小鼠采用小鼠重复轻度脑损伤模型进行造模。通过干湿重比值法检测脑组织水肿、水迷宫实验检测小鼠学习记忆能力、HE染色法观察小鼠脑组织形态学变化、采用免疫组化法检测小胶质细胞的激活情况。结果与TBI组小鼠比较,阿魏酸组小鼠:脑含水量明显降低(P<0.05),Morris水迷宫实验中逃避潜伏期较TBI模型组明显缩短,原象限停留时间明显延长,活化的小胶质细胞减少。结论阿魏酸可改善TBI小鼠的空间学习记忆能力,降低TBI小鼠脑含水量,改善TBI小鼠的脑组织形态学变化,其机制与阿魏酸抑制小胶质细胞活化有关。     

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.     

多指标综合加权评分法优选复方夏枯草洗剂提取工艺研究

[目的] 优选复方夏枯草洗剂的提取工艺。[方法] 采用L₉（3⁴）正交设计，以迷迭香酸、丹酚酸B的含量及出膏率的综合评分为指标，考察料液比、提取时间、提取次数对提取效果的影响，优选复方夏枯草洗剂的最佳提取工艺。[结果] 根据综合评分的结果，优选出最佳提取工艺为加8倍水，提取3次，每次0.5 h。[结论] 该工艺准确可靠，可行性好。     

Serum Folate Levels and Lung Cancer Risk: A Meta- Epidemiological Study of Population-based Case-Control Studies

Objective: While it has been claimed that lung cancer occurs due to epigenetic mechanisms, four systematic reviews were reported to investigate the association between serum folate levels and lung cancer risk. Considering some methodological problems founded in the systematic review, a meta-epidemiological study was conducted. Methods: The selection criteria of this study were defined that a case-control study was conducted to determine the risk of lung cancer occurrence according to the concentration of serum folate and its results showed odds ratio and its 95% confidence interval. Additional paper was explored from cited lists of 4 papers selected by previous systematic reviews. Random effect model was applied if I-squared value was over 50%. Results: For 5 case-control studies selected, the summary odds ratios (and their 95% confidence intervals) were 0.82 (0.74-0.90) in men, 0.70 (0.62-0.79) in former smokers, and 0.86 (0.75-1.00) in non-smokers. Conclusion: Higher foliate levels can decrease lung cancer risk in men and former smokers. Especially, the protective effect was highest in former smokers compared in non-smokers and current smokers. Based on these facts, folate fortification programs to reduce lung cancer risk would be focused on former smokers in men. And some epidemiological studies are needed to provide a hypothesis to explain the sex differences in the association between folate and lung cancer risk.