Metabolism of non-steroidal anti-inflammatory drugs (NSAIDs) by Streptomyces griseolus CYP105A1 and its variants

In the field of drug development, technology for producing human metabolites at a low cost is required. In this study, we explored the possibility of using prokaryotic water-soluble cytochrome P450 (CYP) to produce human metabolites. Streptomyces griseolus CYP105A1 metabolizes various non-steroidal anti-inflammatory drugs (NSAIDs), including diclofenac, mefenamic acid, flufenamic acid, tolfenamic acid, meclofenamic acid, and ibuprofen. CYP105A1 showed 4′-hydroxylation activity towards diclofenac, mefenamic acid, flufenamic acid, tolfenamic acid, and meclofenamic acid. It should be noted that this reaction specificity was similar to that of human CYP2C9. In the case of mefenamic acid, another metabolite, 3′-hydroxymethyl mefenamic acid, was detected as a major metabolite. Substitution of Arg at position 73 with Ala in CYP105A1 dramatically reduced the hydroxylation activity toward diclofenac, flufenamic acid, and ibuprofen, indicating that Arg73 is essential for the hydroxylation of these substrates. In contrast, substitution of Arg84 with Ala remarkably increased the hydroxylation activity towards diclofenac, mefenamic acid, and flufenamic acid. Recombinant Rhodococcus erythrocyte cells expressing the CYP105A1 variant R84A/M239A showed complete conversion of diclofenac into 4′-hydroxydiclofenac. These results suggest the usefulness of recombinant R. erythropolis cells expressing actinomycete CYP, such as CYP105A1, for the production of human drug metabolites.     

The usefulness of measuring n-butyric acid concentration as a new indicator of blood decomposition in forensic autopsy

In forensic medicine, although various alcohols have been reported as indicators of decomposition in collected blood, no studies have examined short-chain fatty acids as indicators. In this study, the blood n-butyric acid concentration was quantified, and the association between n-butyric acid and decomposition was investigated to determine whether the detection of n-butyric acid could be a new indicator of decomposition. Among the forensic autopsies performed from 2016 to 2018 in our laboratory, the cases were divided into decomposed (n = 20) and non-decomposed (n = 20) groups based on macroscopic findings. Blood samples collected at the time of autopsy were derivatized with 3-nitrophenylhydrazine hydrochloride after solid-phase extraction. The n-butyric acid concentration was measured using liquid chromatography–tandem mass spectrometry. In addition, ethanol and n-propanol were measured using a gas chromatography-flame ionization detector. There was a significant difference (p < 0.01) in the concentrations of n-butyric acid between the decomposed and non-decomposed groups (0.343 ± 0.259 [0.030–0.973] and 0.003 ± 0.002 [0.001–0.007] mg/mL, respectively). In the decomposed group, n-butyric acid was detected at high concentrations, even in cases where n-propanol was low. These results suggest that n-butyric acid is more likely to be an indicator of blood decomposition than n-propanol.     

从新型冠状病毒肺炎的寒疫病机探析清肺排毒汤的组方机制

在《新型冠状病毒肺炎诊疗方案(试行第六版)》中推荐的清肺排毒汤,临床观察疗效显著。笔者拟探讨新型冠状病毒肺炎(COVID-19)的寒疫病机以及清肺排毒汤的方药组成机制,认为该方能疏解表里、通调三焦,具有宣肺行气、透邪解毒、润燥化湿、逐水泻热的功效,切合COVID-19寒、燥、湿的病机特点,且能有效的针对症状进行治疗,应推广全国各地广泛使用以遏制疫情蔓延。     

Aberrantly Methylated cfDNA in Body Fluids as a Promising Diagnostic Tool for Early Detection of Breast Cancer

Breast malignancies are the leading type of cancer among women. Its prevention and early detection, particularly in young women, remains challenging. To this end, cell-free DNA (cfDNA) detected in body fluids demonstrates great potential for early detection of tissue transformation and altered molecular setup, such as epigenetic profiles. Aberrantly methylated cfDNA in body fluids could therefore serve as a potential diagnostic and prognostic tool in breast cancer management. Abnormal methylation may lead to both an activation of oncogenes via hypomethylation and an inactivation of tumor suppressor genes by hypermethylation. We update the state of the art in the area of aberrant cfDNA methylation analyses as a diagnostic and prognostic tool in breast cancer, report on the main technological challenges, and provide an outlook for advancing the overall management of breast malignancies based on cfDNA as a target for diagnosis and tailored therapies.     

UPLC-PDA法同时测定复方五凤草液中7种成分

目的建立UPLC-PDA法同时测定复方五凤草液(五凤草、白芨、猫爪草)中没食子酸、原儿茶酸、对羟基苯甲酸、咖啡酸、金丝桃苷、槲皮素、柚皮素的含有量。方法该药物80%甲醇提取液的分析采用Waters BEH C18色谱柱(2.1 mm×100 mm, 1.7μm);流动相乙腈-0.1%甲酸,梯度洗脱;体积流量0.3 mL/min;柱温40℃;检测波长266 nm。结果 7种成分在各自范围内线性关系良好(r≥0.999 5),平均加样回收率96.96%~102.26%,RSD 0.65%~2.10%。结论该方法稳定准确,重复性好,可用于复方五凤草液的质量控制。     

清肺化浊汤联合胸腺五肽治疗恶性肿瘤合并肺部感染效果及对淋巴细胞亚群水平的影响

目的探究清肺化浊汤联合胸腺五肽治疗恶性肿瘤合并肺部感染的临床效果和对淋巴细胞亚群水平影响。方法选取恶性肿瘤合并肺部感染的患者109例作为研究对象,按随机数字表法将其分为两组,对照组54例患者采用胸腺五肽进行治疗,观察组55例患者在观察组的基础上予以清肺化浊汤辅助治疗。观察比较两组治疗前后临床症状改善情况、炎症指标和T淋巴细胞亚群水平变化情况。结果观察组胸部X线好转时间、退热时间、咳痰咳嗽好转时间明显优于对照组(P<0.05);观察组炎症因子水平、CD3^+(71.08±3.86)%、CD4^+(41.17±3.21)%、CD4^+CD25^+(1.52±0.37)%和CD3^+CD+8(25.17±2.56)%水平明显优于对照组炎症因子水平、CD3^+(64.31±4.17)%、CD4^+(34.24±2.53)%、CD4^+CD25^+(2.11±0.45)%和CD3^+CD+8(27.84±2.15)%,差异具有统计学意义(P<0.05)。结论清肺化浊汤联合胸腺五肽治疗恶性肿瘤合并肺部感染患者效果明显,可以有效改善患者的临床症状,降低炎症因子水平,增强患者的免疫功能,值得临床推广应用。     

基于计算机辅助药物设计的清肺排毒汤多靶点系统治疗新型冠状病毒肺炎(COVID-19)物质基础探究

目的通过对清肺排毒汤功效和新型冠状病毒肺炎(COVID-19)病机的分析,基于计算机辅助药物设计(CADD)从多靶点结构出发系统探索清肺排毒汤排毒、抑制炎症风暴、利水渗湿3个方面的物质基础和分子机制。方法 TCMSP下载清肺排毒汤成分,基于血管紧张素转化酶II(ACE2)、白细胞介素-6受体(IL-6R)和水通道蛋白(AQP)分别进行分子对接虚拟筛选并对其结合模式进行分析。对利水药白术、茯苓、猪苓、泽泻进行反向靶点预测和GO分析和KEGG通路富集分析,预测其作用机制。结果阻断病毒作用最突出的前3位中药为甘草、麻黄、枳实,抑制炎症作用最突出的前3位中药是甘草、柴胡、紫菀;清肺排毒汤4个子方中,小柴胡汤阻断病毒、抑制炎症风暴的潜在活性化合物数均排第1。槲皮素及其衍生物对ACE2和IL-6R2个靶点均具有较强结合能力,是潜在的双靶点活性化合物。ACE2的Lys353残基是化合物与ACE2结合的关键位点。白术、茯苓、猪苓、泽泻缺乏阻断病毒、抑制炎症风暴的化合物,分子对接结果显示,这4味药材中含有的东莨菪内酯、去氢齿孔酸、α-D-半乳糖、环氧泽泻烯具有与水通道蛋白4(AQP4)的潜在结合能力。结论清肺排毒汤可通过多种化合物分别与ACE2、IL-6R、AQP4结合,发挥排毒、抑制炎症风暴、利水渗湿的作用;各组成子医方配伍合理,通过多点协同、优势互补发挥防治作用;得到了中药阻断新型冠状病毒(SARS-Co V-2)成分关键的结合位点Lys353,为清肺排毒汤药效机制的多角度挖掘和单体成分的现代化开发提供线索。     

养阴清肺汤对急性放射性口腔炎患者免疫功能、炎症细胞因子的影响

目的观察养阴清肺汤治疗急性放射性口腔炎的临床疗效及其对患者免疫功能和炎症细胞因子的影响。方法患者100例,按照随机数字表法分为治疗组与对照组各50例,两组均采用西医常规治疗,治疗组在对照组治疗基础上加用养阴清肺汤,两组疗程均持续到放疗结束后2~4周。观察两组临床疗效,治疗前后两组唾液中γ-干扰素(IFN-γ)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)及白细胞介素-8(IL-8)水平及两组治疗前后免疫功能指标(CD3^+、CD4^+、CD8^+、CD4^+/CD8^+)的变化情况。结果治疗组总有效率为92.00%,显著高于对照组的76.00%(P<0.05)。治疗组症状体征、唾液炎症细胞因子、相关免疫功能指标恢复程度均优于对照组(P<0.05)。结论养阴清肺汤可有效缓解急性放射性口腔炎的疼痛和黏膜损伤程度,加速患者伤口愈合,有效抑制急性放射性口腔炎的相关炎症细胞因子。