Phytochemical investigation of the roots of Bistorta suffulta (Maxim.) H. Gross led to the isolation of 24 compounds, comprising four triterpenoids (1-4), three flavonoids (5-7), three coumarins (8-10), three sterols (11-13), eight phenolic compounds (14-21), three fatty acids and their derivatives (22-24). Their structures were elucidated through comparative analysis of NMR (1H and 13C) spectroscopic data with literature values. To the best of our knowledge, apart from compound 1 and 11, all isolated compounds are reported from this plant for the first time. Furthermore, 2-4, 8, 9, 12, 13, 16, 19, 23, and 24 are reported for the first time from the Polygonaceae family. The chemotaxonomic significance of these compounds was discussed. Additionally, the growth inhibitory activities of compounds 1-11 and 13 against various cancer cells were evaluated.

Previously undescribed triterpenoid saponins (1-8), along with eight previously reported analogues (9-16), were isolated from an EtOAc extract obtained from the roots of Rubus setchuenensis. Structures were established based on HRESIMS, 1D and 2D NMR, as well as by literature comparisons. All compounds were isolated for the first time from the genus Rubus. Compounds 3-4, 6-8, and 12-15 displayed inhibitory activities against NO production in a RAW264.7 cell inflammation model induced by LPS. Moreover, 4, 6-8, and 12-13 exhibited inhibitory effects in a LPS-induced BV-2 cell neuroinflammation model.

INTRODUCTION:Flavonoids are the main nonvolatile component responsible for the anti-rheumatoid arthritis (RA) synovitis activities of Artemisia argyi Folium. However, the effective ingredient remains unidentified. Spectrum-effect relationships analysis was a reliable and efficient strategy for herbal effective ingredient discovery. OBJECTIVES:This study aimed to screen the effective ingredients within the total flavonoid from Artemisia argyi Folium (TFAA) that exhibit anti-RA synovitis activities based on spectrum-effect relationship. METHODS:TFAA was obtained through ethanol extraction and subsequent purification with D101 resin from 15 distinct batches of Artemisia argyi Folium. The fingerprint of TFAA was established using HPLC, and its efficacy against RA synovitis was evaluated by determining the inhibition rate of nitric oxide (NO) on MH7A synovioblast induced by TNF-α. Common peaks were identified using HPLC-MS/MS and authentic standards. The spectrum-effect relationships between the fingerprints and efficacy were analyzed by gray relational analysis (GRA), canonical correlation analysis (CCA), and partial least squares regression analysis (PLSR) to pinpoint the peaks responsible for the anti-RA synovitis activity, and the results were further verified by in vitro anti-RA synovitis experiments and molecular docking studies. RESULTS:The fingerprint revealed 14 common peaks, and 12 compounds were identified, including four caffeoylquinic acids and eight flavonoids. Among them, five flavonoids-X10 (hispidulin), X11 (jaceosidin), X12 (centaureidin), X13 (eupatilin), and X14 (casticin)-were highly relevant to anti-RA synovitis activity. Verification experiments confirmed their inhibitory effect on NO production and cytokine secretion in MH7A cells, showing anti-RA synovitis potential, which was consistent with the established spectrum effect relationship. The underlying mechanism might be related to the inhibition of iNOS. CONCLUSION:Hispidulin, jaceosidin, centaureidin, eupatilin, and casticin were the key effective ingredient of TFAA responsible for its anti-RA synovitis effects. These compounds can serve as quality control markers for Artemisia argyi Folium and as lead compounds for anti-RA synovitis treatment.

基于网络药理学探究艾叶黄酮治疗类风湿性关节炎(RA)的作用机制.通过 TCMSP 数据库和 STP 数据库并结合文献筛选艾叶活性黄酮成分及其相应靶点,通过 GeneCards、DRUGBANK、OMIM、TTD 数据库预测 RA 相关靶点,得到艾叶黄酮治疗 RA的交集靶点;采用 DAVID 数据库对交集靶点进行 GO 功能富集分析和 KEGG 通路富集分析;并导入STRING数据库进行蛋白相互作用(PPI)分析,筛选出关键靶点;使用 Cytoscape3.7.2 软件构建"成分-靶点-通路"网络,分析并筛选出核心活性成分.筛选获得 38 个艾叶活性黄酮成分,主要作用于 129 个 RA相关靶点;主要通过药物反应、凋亡过程的负调控和炎症应答等生物过程,涉及包括肿瘤坏死因子信号通路、Toll样受体信号通路、破骨细胞分化等 15 条相关的信号通路,关键靶点有 TNF、JUN、RELA、AKT1、TP53、IL-6、MAPK1、MAPK14;核心成分为槲皮素、芹菜素、山柰酚、木犀草素、柚皮素、异鼠李素、异泽兰黄素.表明艾叶黄酮通过多成分、多靶点、多通路协同治疗 RA,为其进一步实验研究和临床应用提供依据.

In recent years, the biopharmaceutical industry has developed rapidly, creating urgent demand for high-quality, innovative, and application-oriented talents. In the context of "first-class undergraduate education", it is of great significance to reform and explore biopharmaceutics blended learning to foster professional talents who can adapt to the industrial development. The blended teaching of biopharmaceutics course in Hubei University was based on small private online course (SPOC) and ChaoXing platform, aiming to meet the first-class "AIC (advanced, innovation, challenge)". The course strengthened the three phases of teaching: before, during, and after class, and innovated teaching methods actively to achieve curriculum goals, and integrated typical cases organically. In addition, the course improved the discriminative power of assessment by strengthening the formative performance evaluation. Moreover, the course provided guidance for students to improve the learning efficiency through investigating the students' learning behavior and employing the marginal utility curve to analyze the characteristics of group activities. Furthermore, the course also offered students personalized learning guidance based on their career planning. The reform of biopharmaceutics blended teaching has achieved significant outcomes, such as improving students' satisfaction, students' innovation and entrepreneurship ability, and curriculum construction level, thus may serve as a reference for the teaching reform and research of the related courses.

Three neolignan glycosides, including a new compound (7S,8R)‐dihydro‐3′‐hydroxy‐7‐(4‐hydroxy‐3‐methoxyphenyl)‐1′‐benzofuranpropanol‐9‐O‐β‐D‐xylopyranoside (1), were isolated from the root of Nothopanax davidii. Their structures were determined by extensive spectroscopic analyses, particularly NMR, HR‐ESI‐MS, and ECD experiments, and the absolute configuration of 2 was first definitively determined. The anti‐tumor activity was assessed on four tumor cells by MTT assay, the anti‐inflammatory activity was determined by inhibition of NO production in LPS reduced RAW264.7 cells, and the interaction with iNOS was predicted by molecular docking. At the dose of 100 μM, the three neolignan glycosides showed no cytotoxic activity against HepG2, HCT116, HeLa and A549 human tumor cells, but significantly inhibited LPS induced NO generation in RAW264.7 cells with inhibition rates of 31.53 %, 23.95 %, and 20.79 %, respectively, showing weak anti‐inflammatory activity, possibly due to their binding to key residues of iNOs involved in inhibitor binding.

Beta-sitosterol (1) and ingol 7,8,12-triacetate 3-phenylacetate (2) were isolated from leaves of Synadenium grantii Hook F. and screened for anti-tumor activity against a NCI60 cell line panel. The highly functionalized ingol diterpene (2) displayed cytotoxicity against leukemia cancer cells (SR) and renal cancer cells (CAKI-1) with growth inhibition of 33% and 21%, respectively. Since PIK3α inhibitors are predicted to target PI3K/Akt signaling that is operative in renal cancer and leukemia, in silico molecular docking simulations with an alpha isoform of PIK3 were performed. Docking simulations confirmed that 2 fits in the active site with a docking score of −9.03 kcal/mol. Computational modeling included biological validation against the enzyme target, PI3KCα based on the commercial inhibitors X6K and GDC-0326 with experimental IC50 values of 18 and 0.2 nM, respectively. Ingol diterpene (2), isolated from the medicinal herb S. grantii, holds potential as an inhibitor for select tumors; binding simulations indicate possible inhibition via the PI3K/Akt signaling pathway.

β-Nicotinamide mononucleotide (NMN) has been widely used as a nutraceutical for self-medication. The one-step conversion of nicotinamide riboside (NR) to β-NMN has been considered to be the most promising synthetic route for β-NMN. Here, human nicotinamide riboside kinase 2 (NRK-2) was functionally displayed on the cell surface of Saccharomyces cerevisiae EBY100, forming a whole-cell biocatalyst (Whole-cell NRK-2). Whole-cell NRK-2 could convert nicotinamide riboside (NR) to β-NMN efficiently in the presence of ATP and Mg2+, with a maximal activity of 64 IU/g (dry weight) and a Km of 3.5 μM, similar to that of free NRK-2 reported previously. To get the best reaction conditions for β-NMN synthesis, the amounts of NR, ATP, and Mg2+ used, pH, and temperature for the synthetic reaction were optimized. Using Whole-cell NRK-2 as the catalyst under the optimized conditions, β-NMN synthesized from NR reached a maximal conversion rate of 98.2%, corresponding to 12.6 g/L of β-NMN in the reaction mixture, which was much higher than those of synthetic processes reported. Additionally, Whole-cell NRK-2 had good pH stability and thermostability, required no complicated treatments before or after use, and could be reused in sequential production. Therefore, this study provided a safe, stable, highly effective, and low-cost biocatalyst for the preparation of β-NMN, which has great potential in industrial production.

Non-alcoholic fatty liver disease (NAFLD) is a prevalent chronic liver disease associated with hepatic lipid accumulation, insulin resistance, oxidative stress, and inflammation. The CFLAR-JNK pathway plays a decisive role in the development of NAFLD. EJ1 is a new hexacyclic triterpenic acid isolated from Euscaphis japonica, a traditional herbal medicine with anti-NAFLD effect. The present study was aimed to evaluate the hepatoprotective effect of EJ1 against NAFLD in vitro and the underlying mechanisms. The biochemical parameters related to lipid accumulation, insulin resistance, oxidative stress, and inflammation in oleic acid-induced HepG2 cellular model of NAFLD were measured. In this study, EJ1 was found to significantly decrease TG and TC contents. Meanwhile, EJ1 increased hepatocellular glucose uptake and improved insulin resistance by increasing the phosphorylation levels of pIRS1, pAKT and PI3K. Furthermore, EJ1 decreased the intracellular ROS and MDA, promoted the antioxidant enzymes activity including SOD and GSH-Px. Moreover, EJ1 reduced the levels of cellular NO and mRNA of TNF-α, IL-6 and IL-8. Further investigation showed that EJ1 treatment led to up-regulation of CFLAR protein expression and down-regulation of JNK protein phosphorylation. These results revealed that EJ1 has the potential to alleviate lipid accumulation, insulin resistance, oxidative stress, and inflammatory responses in OA-treated HepG2 cellular model of NAFLD, representing a promising leading compound for NAFLD, possibly by activating the CFLAR-JNK pathway.

目的 研究华中枸骨Ilex centrochinensis根的化学成分.方法 运用硅胶、Sephadex LH-20,ODS柱色谱及半制备HPLC等方法进行分离纯化,并通过NMR等波谱技术和理化性质鉴定结构.结果 从华中枸骨根中分离得到13个化合物,分别鉴定为11-羰基-α-香树醇-3β-棕榈酸酯(1)、3β-乙酰氧基-28-羟基-12-烯-乌苏烷(2)、儿茶酚(3)、4-乙基儿茶酚(4)、2,4,6-三甲氧基苯酚(5)、3-(4-羟基-3,5-二甲氧基苯基)-1,2-丙二醇(6)、3-(4-羟基-3-甲氧基苯基)-1,2-丙二醇(7)、3,4-二甲氧基苯酚(8)、3,4-二羟基苯丙酸甲酯(9)、3β,19α,23-三羟基-20α(H)-乌苏-12-烯-28-酸3β-O-α-L-吡喃阿拉伯糖(10)、(+)-1-羟基松脂醇-1-O-β-D-葡萄糖苷(11)、3β-O-[α-L-吡喃鼠李糖(1→2)-α-L-吡喃阿拉伯糖]-坡模酸(12)、rotundioic acid (13).结论 所有化合物均为首次从华中枸骨中分离得到,其中4～9为首次从冬青属植物中分离得到.

Highly specific novel glycopeptide-based fluorescent molecularly imprinting polymers (g-FMIPs) were constructed to recognize and determine the target glycoprotein in complex biological samples. The glycopeptide of ovalbumin (OVA), with the unique structural characteristics of glycan and peptide, and potential application in improving the specificity recognition of g-FMIPs, was selected as the template molecule. The nitrogen-doped graphene quantum dots (N-GQDs) were introduced for fluorescence response. The obtained g-FMIPs possessed rapid binding kinetics and high adsorption capacity. Notably, the g-FMIPs exhibited remarkable selectivity and sensitivity with a high imprinting factor of 6.57, good linearity of 0.625 − 5.00 μM, and limit of detection of 0.208 μM. After treatment with g-FMIPs, the concentration of OVA in eluted solution was 1.07 μM. The obtained recoveries at 1.43 μM, 2.86 μM, and 4.29 μM spiked concentrations were 97.2%, 93.5%, and 101%, respectively, and the relative standard deviations were 2.6%, 4.2%, and 1.1%, respectively. In summary, the proposed strategy will expand the MIPs construction method and its application prospects in precision recognition and sensitive detection of trace glycoproteins from complex biosamples.

以总黄酮含量和转移率为评价指标,采用多种方法提取纯化华中枸骨叶总黄酮(ICTF),并以TNF-α诱导人关节滑膜成纤维细胞MH7A为滑膜炎症细胞模型评价其抗RA滑膜炎症活性,试剂盒测定炎症介质NO含量,RT-qPCR检测炎症因子IL-1β、IL-6、IL-8mRNA的转录水平.结果表明,华中枸骨叶乙醇总提取物经石油醚脱脂后,经D101大孔吸附树脂吸附,50％乙醇洗脱5 BV,再经葡聚糖凝胶Sephadex LH-20吸附后甲醇洗脱5 BV后,所得总黄酮含量达53.91％,转移率为39.53％;所得ICTF能剂量依赖性降低滑膜炎症模型细胞NO水平以及IL-1βmRNA的转录水平,200μg/mL剂量可显著降低IL-6和IL-8 mRNA的水平.该纯化方法稳定可行,所得ICTF具有抑制RA滑膜炎症潜力.

One new siaresinolic acid saponin (1) and three new rotundic acid saponins (2-4) were isolated from the roots of Ilex centrochinensis. Their structures were confirmed by detailed analysis of standard spectroscopic data (IR, MS, 1D and 2D NMR). Compounds 1-4 exhibited anti-inflammatory activity by inhibiting nitric oxide production in a lipopolysaccharide-induced RAW264.7 cell inflammatory model. However, they showed no significant lipid-lowering activity against the production of triglycerides in the lipid-accumulation model of HepG2 cells induced by oleic acid.

Objective To provide an alternative therapeutic modality for rheumatoid arthritis (RA), a novel bispecific antibody (BsAb) targeting human tumor necrosis factor α (TNF-α) and human complement component C5a was constructed. Results BsAb was expressed in Pichia pastoris and secreted into the culture medium as a functional protein. In vitro functional study demonstrated that BsAb could simultaneously bind to TNF-α and C5a and neutralize their biological actions. Furthermore, BsAb showed significant improvements in both the antigen-binding affinity and the neutralizing ability as compared to its original antibodies produced in E. coli . It was also found that TNF-α and C5a had an additive/synergistic effect on promoting the production of inflammatory cytokines and chemokines and C5a receptor (C5aR) expression in human macrophages. Compared to single inhibition of TNF-α or C5a with respective antibody, BsAb showed a superior efficacy in blocking inflammatory cytokines, chemokines, and C5aR response, as well as in lowering the C5a-mediated chemotaxis of macrophages via C5aR in vitro. Conclusions With improved production processing and the ability to simultaneously block TNF-α and C5a action, BsAb has a great potential to be developed into a therapeutic agent and may offer a better therapeutic index for RA.

A pair of new diastereoisomeric flavan, containing an additional phenylpropanoid (C6–C3) unit in the molecule, has been isolated from the leaves of Ilex centrochinensis. Their structures were identified by extensive spectral analysis and comparison with the data of known analogues.

为研究彩云木Synadenium grantii叶的化学成分及部分化合物体外抗炎活性,本研究采用硅胶、Sephadex LH-20等多种柱色谱进行分离纯化,通过理化性质和NMR等波谱数据鉴定化合物的结构,并对部分化合物进行抑制脂多糖诱导RAW 264.7细胞释放NO活性的测定.从彩云木叶二氯甲烷-甲醇(1∶1)提取物中共分离得到了10个化合物,分别鉴定为日尔曼醇(1)、大戟醇(2)、正二十八烷醇(3)、β-谷甾醇(4)、豆甾醇(5)、ingol-7,8,12-triacetate-3-phenylacetate (6)、山柰酚-3-O-α-L-吡喃鼠李糖苷(7)、胡萝卜苷(8)、3,4,3'-三甲氧基鞣花酸-4'-O-β-D-吡喃葡萄糖苷(9)、山奈酚-3-O-β-D-吡喃葡萄糖苷(10).其中3和6～10为首次从该种植物中分离得到,3、6、8和9为首次从聚苞大戟属植物中分离得到,9抑制LPS诱导RAW 264.7细胞释放NO的IC50值为12.0±0.9 μM,显示较好的体外抗炎活性.

Herpes simplex encephalitis (HSE) is the most common infectious disease of the central nervous system worldwide. However, the pathogenesis of HSE is not clear. Research has shown that the immune response mediated by the toll-like receptor 3 (TLR3) signaling pathway is essential to protect the central nervous system against herpes simplex virus (HSV) infection. However, an excessive immune response may cause tissue damage accompanied by pathological changes. The aim of this study was to explore the molecular mechanism via which corilagin controls HSE through the TLR3 signaling pathway in vitro and in vivo. Cells and mice were pre-treated with polyriboinosinic polyribocytidylic acid [poly(I:C)] or HSV type 1, and then treated with corilagin. After treatment, the mRNA and protein levels of TLR3, TLR-like receptor-associated interferon factor (TRIF), tumor necrosis factor (TNF) receptor type 1-associated DEATH domain protein (TRADD), TNF receptor-associated factor (TRAF) 3 and 6, nuclear factor-kappa-B (NF-κB) essential modulator (NEMO), P38, and interferon regulatory factor 3 (IRF3) were decreased. Interleukin-6 (IL-6), TNF-α, and type 1 interferon-β were also decreased. When TLR3 expression was silenced or increased, corilagin still inhibited the expression of TLR3 and its downstream mediators. Hematoxylin-eosin (HE) staining and immunohistochemical examinations of mouse brain tissues revealed that corilagin lessened the degree of brain inflammation. Altogether, these results suggest that corilagin may regulate the immune response in HSE and relieve inflammatory injury by interfering with the TLR3 signaling pathway.

Inflammatory cytokines can mediate many biological processes and are tightly regulated by the body. Loss of control can trigger a range of diseases such as autoimmune inflammation and cancer. Therefore, a number of biological agents that can effectively regulate the biological effects of inflammatory cytokines such as recombinant anti-inflammatory cytokines, cytokine receptors and neutralizing antibodies have been extensively used in the treatment of related diseases caused by the imbalance of inflammatory cytokines. In recent years, in particular, a number of new innovative biological agents for blocking and regulating cytokine activities are emerging. In this article, we review the recent development and clinical use of the biologics targeting TNF-α, IL-1, IL-6 and IL-17, and point out their inherent limitations and clinical risks. Finally, based on the research findings of our own and other scholars, we suggest some approaches and methods for reducing their side-effects and clinical risk. We consider that using modern biotechnology to improve the tissue specificity to inflammatory site and tumor will be an important development direction of such biologics.

A new para-quinone-type flavan, (2S)-7-methoxy-3',4'-dihydroxy-5,8-quinoflavan (1), together with three known compounds, were isolated from the leaves of Ilex centrochinensis. Their structures were elucidated by detailed spectroscopic analyses for new structure and in comparison with published data for known compounds. Moreover, the new compound was evaluated its cytotoxic and anti-inflammatory activities in vitro on LPS induced RAW 264.7 cells and the results showed that 1 has promising anti-inflammatory activities.

Objective To screen the active fraction of Euscaphisjaponica roots with inhibitory activity on hepatic lipid accumulation and investigate its chemical constituents.Methods Different polar fractions were prepared by extraction with organic solvents,oil Red O staining and triglyceride content assay were used to determine inhibitory activity of different polar fraction on oleic acid induced triglyceride accumulation on HepG2 cells,and the constituents of active fraction were isolated and purified by various chromatography techniques such as column chromatography on silica gel,Sephadex LH-20,and HPLC,and their structures were identified by physicochemical properties and spectral data.Results The petroleum ether fraction exhibited significantly inhibitory activity on oleic acid induced triglyceride accumulation on HepG2 cells.Nine compounds were isolated and identified as 3β,19-dihydroxy-24-trans-ferulyloxyurs-12-en-28-oic acid (1),β-sitosterol (2),7-hydroxy-2-octen-5-olide (3),3,3'-dimethoxy-ellagic acid (4),vanillin (5),ethyl-5-oxo-tetrahydro-3-furancarboxylate (6),gallic acid (7),3,3'-di-O-methylellagic acid 4-(5"-acetyl)-α-L-arabinofuranoside (8),and bergapten (9).Conclusion The petroleum ether fraction is main active fraction.The compounds 1,6,8,and 9 are obtained from genus Euscaphis Sieb.et Zucc.for the first time.