皖产古尼虫草中核苷和碱基成分的HPLC分析

目的:建立高效液相色谱法测定皖产古尼虫草中核苷和碱基成分(尿嘧啶、虫草素、腺苷、腺嘌呤、尿苷、次黄嘌呤、肌苷和鸟苷)的含量,并比较其与天然冬虫夏草核苷和碱基成分的异同。方法:采用Zorbax SB-Aq(250 mm×4.6 mm,5μm)色谱柱,以水-乙腈为流动相,梯度洗脱,流速为0.8 mL.min-1,检测波长260 nm,柱温20℃。结果:尿嘧啶、虫草素、腺苷、腺嘌呤、尿苷、次黄嘌呤、肌苷和鸟苷进样浓度分别在0.38～191.08,0.37～188.48,0.37～189.72,0.37～190.92,0.37～188.55,0.50～168.55,0.37～190.36,0.24～125.00μg.mL-1范围内线性关系良好,R2为0.9998～0.9999;加样回收率分别为102.1%,102.1%,95.3%,96.2%,96.9%,94.1%,108.7%,101.8%,RSD未超过2.8%。皖产古尼虫草和冬虫夏草均含有核苷和碱基成分,其中:皖产古尼虫草含有较高的尿嘧啶、尿苷、腺苷,而冬虫夏草中肌苷含量较高;二者的次黄嘌呤、鸟苷、腺嘌呤的含量相近;在该检测限下,两者均未测得虫草素。结论:初步认...     

Targeting a cryptic allosteric site of SIRT6 with small-molecule inhibitors that inhibit the migration of pancreatic cancer cells

SIRT6 belongs to the conserved NAD⁺-dependent deacetylase superfamily and mediates multiple biological and pathological processes. Targeting SIRT6 by allosteric modulators represents a novel direction for therapeutics, which can overcome the selectivity problem caused by the structural similarity of orthosteric sites among deacetylases. Here, developing a reversed allosteric strategy AlloReverse, we identified a cryptic allosteric site, Pocket Z, which was only induced by the bi-directional allosteric signal triggered upon orthosteric binding of NAD⁺. Based on Pocket Z, we discovered an SIRT6 allosteric inhibitor named JYQ-42. JYQ-42 selectively targets SIRT6 among other histone deacetylases and effectively inhibits SIRT6 deacetylation, with an IC₅₀ of 2.33 μmol/L. JYQ-42 significantly suppresses SIRT6-mediated cancer cell migration and pro-inflammatory cytokine production. JYQ-42, to our knowledge, is the most potent and selective allosteric SIRT6 inhibitor. This study provides a novel strategy for allosteric drug design and will help in the challenging development of therapeutic agents that can selectively bind SIRT6.     

Metabolic dysregulation and emerging therapeutical targets for hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is an aggressive human cancer with increasing incidence worldwide. Multiple efforts have been made to explore pharmaceutical therapies to treat HCC, such as targeted tyrosine kinase inhibitors, immune based therapies and combination of chemotherapy. However, limitations exist in current strategies including chemoresistance for instance. Tumor initiation and progression is driven by reprogramming of metabolism, in particular during HCC development. Recently, metabolic associated fatty liver disease (MAFLD), a reappraisal of new nomenclature for non-alcoholic fatty liver disease (NAFLD), indicates growing appreciation of metabolism in the pathogenesis of liver disease, including HCC, thereby suggesting new strategies by targeting abnormal metabolism for HCC treatment. In this review, we introduce directions by highlighting the metabolic targets in glucose, fatty acid, amino acid and glutamine metabolism, which are suitable for HCC pharmaceutical intervention. We also summarize and discuss current pharmaceutical agents and studies targeting deregulated metabolism during HCC treatment. Furthermore, opportunities and challenges in the discovery and development of HCC therapy targeting metabolism are discussed.     

RP-HPLC测定天然虫草与人工虫草中核苷类成分的含量

 目的分析不同来源的天然虫草与人工虫草中主要核苷类成分的差异。方法应用RP-HPLC检测18个样品中尿苷、腺嘌呤、腺苷、虫草素的含量。Symmetry Shield Rp18柱(3.9 mm×150 mm,5μm),以乙腈-水为流动相,梯度洗脱。流速0.2～1.2mL·min^-1,检测波长260 nm,柱温30℃。结果本法高效灵敏,重现性好。所测人工虫草核苷类成分的含量较天然虫草高,不同产地天然虫草4种主要的核苷类成分的差异较不同来源人工虫草的小。结论本法能有效鉴别虫草的核苷类成分,可用于天然虫草和人工虫草中核苷类成分的分析,评价不同来源虫草的质量。     

Prognostic value of nicotinamide adenine dinucleotide (NAD+) metabolic genes in patients with stomach adenocarcinoma based on bioinformatics analysis

BackgroundBecause stomach adenocarcinoma (STAD) has a poor prognosis, it is necessary to explore new prognostic genes to stratify patients to guide existing individualized treatments.MethodsSurvival and clinical information, RNA-seq data and mutation data of STAD were acquired from The Cancer Genome Atlas (TCGA) database. Fifty-one nicotinamide adenine dinucleotide (NAD⁺) metabolism-related genes (NMRGs) were obtained from the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Reactome databases. Differentially expressed NMRGs (DE-NMRGs) between STAD and normal samples were screened, and consistent clustering analysis of STAD patients was performed based on the DE-NMRGs. Survival analysis, Gene Set Enrichment Analysis (GSEA), mutation frequency analysis, immune microenvironment analysis and drug prediction were performed among different clusters. Additionally, the differentially expressed genes (DEGs) among different clusters were selected, and the intersections of DEGs and DE-NMRGs were selected as the prognostic genes. Finally, quantitative real-time polymerase chain reaction (qRT-PCR) was performed on a human gastric mucosa epithelial cell line and cancer cell line to verify the expression of the prognostic genes.ResultsA total of 27 DE-NMRGs and two clusters were selected. There was a difference in survival between clusters 1 and 2. Furthermore, 18 DE-NMRGs were significantly different between clusters 1 and 2. The different Gene Ontology (GO) biological processes and KEGG pathways between clusters 1 and 2 were mainly enriched in cyclic nucleotide mediated signaling, synaptic signaling and hedgehog signaling pathway, etc. The somatic mutation frequencies were different between the two clusters, and TTN was the highest mutated gene in the patients of the clusters 1 and 2. Additionally, eight immune cells, immune score, stromal score, and estimate score were different between clusters 1 and 2. The patients in cluster 2 were sensitive to CTLA4 inhibitor treatment. Furthermore, the top five drugs (AP.24534, BX.795, Midostaurin, WO2009093927 and CCT007093) were significantly higher in cluster 1 than in cluster 2. Finally, three genes (AOX1, NNMT and PTGIS) were acquired as prognostic, and their expressions were consistent with the results of bioinformatics analysis.ConclusionsThree prognostic genes related to NAD⁺ metabolism in STAD were screened out, which provides a theoretical basis and reference value for future treatment and prognosis of STAD.     

线粒体DNA 5178 A/C多态性与2型糖尿病肾病相关性

目的探讨线粒体DNA 5178A／C多态性与2型糖尿病肾病（DN）的相关性。方法将1039例2型糖尿病患者分为无糖尿病肾病（DN-0）组和糖尿病肾病（DN）组,后者进一步分为微量蛋白尿（DN-1）组和大量蛋白尿（DN-2）组,进行病例对照研究。采用聚合酶链反应-限制性片段（PCR-RFLP）技术检测线粒体DNA5178A／C多态。结果（1）2型糖尿病者尿白蛋白肌酐比值（ACR）与5178A／C多态无相关。（2）DN-0,DN-1及DN-2这3组间基因型频率的差异无显著性。结论线粒体DNA 5178A／C多态性与中国2型糖尿病患者ACR水平无相关,与糖尿病肾病无相关。     

Exploration of 5-cyano-6-phenylpyrimidin derivatives containing an 1,2,3-triazole moiety as potent FAD-based LSD1 inhibitors

Histone lysine specific demethylase 1 (LSD1) has become a potential therapeutic target for the treatment of cancer. Discovery and develop novel and potent LSD1 inhibitors is a challenge, although several of them have already entered into clinical trials. Herein, for the first time, we reported the discovery of a series of 5-cyano-6-phenylpyrimidine derivatives as LSD1 inhibitors using flavin adenine dinucleotide (FAD) similarity-based designing strategy, of which compound 14q was finally identified to repress LSD1 with IC₅₀ = 183 nmol/L. Docking analysis suggested that compound 14q fitted well into the FAD-binding pocket. Further mechanism studies showed that compound 14q may inhibit LSD1 activity competitively by occupying the FAD binding sites of LSD1 and inhibit cell migration and invasion by reversing epithelial to mesenchymal transition (EMT). Overall, these findings showed that compound 14q is a suitable candidate for further development of novel FAD similarity-based LSD1 inhibitors.     

Impact of obese levels on the hepatic expression of nuclear receptors and drug-metabolizing enzymes in adult and offspring mice

The prevalence of obesity-associated conditions raises new challenges in clinical medication. Although altered expression of drug-metabolizing enzymes (DMEs) has been shown in obesity, the impacts of obese levels (overweight, obesity, and severe obesity) on the expression of DMEs have not been elucidated. Especially, limited information is available on whether parental obese levels affect ontogenic expression of DMEs in children. Here, a high-fat diet (HFD) and three feeding durations were used to mimic different obese levels in C57BL/6 mice. The hepatic expression of five nuclear receptors (NRs) and nine DMEs was examined. In general, a trend of induced expression of NRs and DMEs (except for Cyp2c29 and 3a11) was observed in HFD groups compared to low-fat diet (LFD) groups. Differential effects of HFD on the hepatic expression of DMEs were found in adult mice at different obese levels. Family-based dietary style of an HFD altered the ontogenic expression of DMEs in the offspring older than 15 days. Furthermore, obese levels of parental mice affected the hepatic expression of DMEs in offspring. Overall, the results indicate that obese levels affected expression of the DMEs in adult individuals and that of their children. Drug dosage might need to be optimized based on the obese levels.