Osteosarcoma is a malignant bone tumor that occurs in adolescents and for which surgical resection and chemotherapy are the treatments of first choice. However, tumor cell metastasis and the toxic effects of chemotherapeutic agents bring great challenges to the treatment of osteosarcoma. Fisetin, a dietary flavonoid derived from vegetables and fruits, plays a therapeutic role in many cancers, but its role and underlying mechanism on osteosarcoma remain unrevealed. This study found that fisetin significantly inhibited the proliferation of osteosarcoma cells and organoids. RNA sequencing revealed that forkhead box O3 (FOXO3) may be a key target of fisetin in its anti-osteosarcoma activity, which was also verified by immunofluorescence. siFOXO3 reversed the effects of fisetin on proliferation, apoptosis, and migration of osteosarcoma cells. Fisetin exerts its effects by targeting the ROS/FOXO3 pathway to promote the binding of FOXO3 to the microtubule-associated protein light chain 3 (LC3) promoter, thereby upregulating LC3 levels and inducing autophagy. Subsequently, activated autophagy further triggers ferroptosis, while silencing LC3 reversed fisetin-induced ferroptosis. In vivo, fisetin inhibited tumor growth while promoting FOXO3 and LC3, and inhibiting SLC7A11 protein expression. Therefore, fisetin-induced autophagy in osteosarcoma cells may be associated with the reactive oxygen species (ROS)/FOXO3 axis, thereby promoting ferroptosis. These findings highlight fisetin's potential as a unique therapy for osteosarcoma.

The chemical constituents in Urtica dioica subsp.gansuensis were isolated systematically and identified by preparative high performance liquid chromatography(HPLC),nuclear magnetic resonance(NMR),high-resolution mass spectrometry(HRMS),and other methods.Their anti-inflammatory activities were evaluated in vitro.As a result,33 compounds were isolated and identified as(7R,8S,8'R)-olivil-4-O-β-D-glucopyranoside(1),oleanic acid(2),vanillic acid(3),L-ascorbic acid(4),p-hydroxybenzaldehyde(5),rosmarinic acid(6),p-hydroxybenzoic acid(7),salicylic acid(8),ficusal(9),lumichrome(10),lobechine(11),2-[formyl-5-(butyl ether methyl)-1H-pyrrol-1-yl]butyric acid butyl ester(12),2-[formyl-5-(methoxymethyl)-1H-pyrrol-1-yl]butyric acid methyl ester(13),2-[formyl-5-(methoxymethyl)-1H-pyrrol-1-yl]butyric acid butyl ester(14),benzoic acid(15),roseoside(16),benzyl-O-β-D-glucopyranoside(17),arbutin(18),carvone(19),trans-pinocarveol(20),carvacrol(21),taxifolin(22),pinocembrin(23),isovitexin(24),3,4-divanillyltetrahydrofuran(25),apigenin-7-O-β-D-glucuronide(26),apigenin(27),β-ionone(28),trans-anethole(29),apigenin-7-O-β-D-glucopyranoside(30),afzelin(31),luteolin(32),and dehydrovomifoliol(33).Compound 1 was a new compound.Various phenolic acids(6-8,and 18)and alkaloids(11-14)exhibited significant anti-inflammatory activities and could significantly inhibit lipopolysaccharide(LPS)-induced NO release from RAW264.7 cells.

ETHNOPHARMACOLOGICAL RELEVANCE:Marsdenia tenacissima extract (MTE) from the stem of the Traditional Chinese herbal medicine of Marsdenia tenacissima (Roxb.) Wight et Arn. has been used as an anticancer remedy for decades. AIM OF THE STUDY:To investigate the beneficial effects of MTE in osteosarcoma in vitro and in vivo and its potential mechanisms. MATERIAL AND METHODS:Nude mice with ectopic xenograft tumors were treated with MTE of 10 mg/kg and 20 mg/kg for 2 weeks. Human osteosarcoma (OS) cell lines 143B and MG63 were treated with 40, 60, and 80 mg/mL MTE for 24 h. RESULTS:MTE significantly inhibited OS cell proliferation in vitro and in vivo. Specifically, intracellular ferrous ion accumulation is closely related to the upregulation of HO-1 expression levels regulated by MTE. NAC and DFO could restore the effects of MTE on OS cell viability. In addition, the Fe2+ accumulation caused cellular oxidative stress and promoted HO-1 translocation to mitochondria. Correspondingly, the mitochondria were damaged, lysosomes were recruited around, and the expression of autophagy-related proteins PINK1 and Parkin was upregulated. Moreover, the autophagy inhibitor 3-MA reversed the effect of MTE on HO-1 and GPX4 protein expression levels in OS cells. In vivo experiments demonstrated that MTE inhibited the growth of xenograft osteosarcoma while increasing the expression of LC3B, HO-1 and FTH1, and simultaneously suppressing the level of GPX4 in tumor tissues. CONCLUSION:MTE exerts anti-osteosarcoma effects by modulating HO-1 and activating mitophagy, thereby accelerating the ferroptosis.

The seed kernel of Momordica cochinchinensis,i.e.,Momordicae Semen,is used for medicinal purposes,but to date,no research has been reported on its chemical constituents.In this study,the chemical constituents of Momordicae Semen were investigated for the first time using silica gel column chromatography,semi-preparative HPLC,HR-MS,and NMR.As a result,eight compounds were isolated and identified as:p-hydroxybenzoic acid-7-O-trehaloside(mubeside A,1),2,6-dimethoxyphenol-O-β-D-apiosyl-(1→2)-β-D-glucoside(mubeside B,2),1-O-p-methoxybenzoyl-1,4-benzenediol-4-O-β-D-apiosyl-(1→2)-β-D-glucoside(mubeside C,3),1-O-p-hydroxybenzoyl-1,4-benzenediol-4-O-β-D-apiosyl-(1→2)-β-D-glucoside(mubeside D,4),gypsogenin-3-O-β-D-galactosyl-(1→2)-β-D-glucuronoside(5),quillaic acid-3-O-β-D-galactosyl-(1→2)-β-D-glucuronoside(6),violanthin(7),and kaempferitrin(8).Compounds 1-4 are new compounds,while compounds 5-8 were isolated from Momordicae Semen for the first time.

Objective The main objectives of this study were to identify the active components of Tongguanteng injection (TGT) and investigate the preclinical efficacy and mechanism of TGT on osteosarcoma using a combination of network pharmacology and experimental validation. Methods To identify the active constituents and targets of TGT against osteosarcoma using network pharmacology, we constructed a network consisting of an 'active ingredient-disease-target-pathway' and a protein–protein interaction (PPI) network. The target organ network was utilized to investigate the distribution of core targets in tissues. Afterwards, the core targets underwent Gene ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. The binding energy between receptors and ligands was compared using molecular docking. In addition, SwissADME was employed to forecast the pharmacokinetic characteristics of the substances. Finally, real-time polymerase chain reaction (RT-PCR), cell proliferation assay, morphological analysis, apoptosis assay, mitochondrial membrane potential (MMP) detection, and Western blotting were utilized to confirm the potential mechanisms of TGT treatment in osteosarcoma cell lines 143B and SAOS2. Results A total of 54 chemical constituents of TGT and 71 targets associated with osteosarcoma were acquired. Through the molecular docking technology, Tenacigenin B, Marsdekoiside, Taraxasterol, Tenacissoside G, Tenacissoside L, and Tenacissoside J were identified as the primary active components of TGT among the various compounds. Analysis of target organs suggests that TGT may play an anti-osteosarcoma role through immune regulation. The GO and KEGG enrichment analysis revealed that TGT could trigger osteosarcoma cell apoptosis by inhibiting the HIF-1 signalling pathway and modulating PD-1 expression and the PD-1 checkpoint pathway in cancer. SwissADME database predicted that Tenacigenin B and Taraxasterol had the best drug-likeness. In vitro studies also demonstrated that TGT suppressed the activity and induced alterations in the morphology of osteosarcoma cells. It decreased MMP levels, triggered apoptosis by increasing Bax expression and Caspase-3 activity, and decreased Bcl-2 expression, thereby exerting an anti-osteosarcoma effect. In the meantime, RT-PCR tests demonstrated that TGT could control immune response against tumors and hinder the proliferation and spread of cancerous cells by impacting the levels of critical factors, including JUN, HSP90AA1, HDAC1, and CDK1. Conclusion The study accurately anticipated the active components, targets, and pathways of TGT in the management of osteosarcoma. The molecular mechanism of TGT-induced apoptosis in osteosarcoma cells was demonstrated by in vitro experiments. These results provide theoretical and technical support for TGT as a clinical adjuvant drug for osteosarcoma.

ETHNOPHARMACOLOGICAL RELEVANCE:As a classical traditional Chinese medicine formula to invigorating spleen and replenishing qi, Sijunzi decoction (SJZD) is composed of four herbs, which is applied to cure spleen deficiency syndrome (SDS) clinically. The non-polysaccharides (NPSs) of SJZD (SJZD_NPS) are important pharmacodynamic material basis. However, the amelioration mechanism of SJZD_NPS on SDS has not been fully elaborated. Additionally, the contribution of herbs compatibility to efficacy of this formula remains unclear.AIM OF THE STUDY:The aim was to explore the underlying mechanisms of SJZD_NPS on improving SDS, and uncover the scientific connotation in SJZD compatibility.MATERIALS AND METHODS:A strategy integrating incomplete formulae (called "Chai-fang" in Chinese) comparison, pharmacodynamics, gut microbiome, and metabolome was employed to reveal the role of each herb to SJZD compatibility against SDS. Additionally, the underlying mechanism harbored by SJZD_NPS was further explored through targeted metabolomics, network pharmacology, molecular docking, pseudo-sterile model, and metagenomics.RESULTS:SJZD_NPS significantly alleviated diarrhea, disordered secretion of gastrointestinal hormones and neurotransmitters, damage of ileal morphology and intestinal barrier in SDS rats, which was superior to the NPSs of Chai-fang. 16S rRNA gene sequencing and metabolomics analyses revealed that SJZD_NPS effectively restored the disturbed gut microbiota community and abnormal metabolism caused by SDS, showing the most evident recovery. Moreover, SJZD_NPS recalled the levels of partial amino acids, short chain fatty acids and bile acids, which possessed strong binding affinity towards potential targets. The depletion of gut microbiota confirmed that the SDS-amelioration efficacy of SJZD_NPS is dependent on the intact gut microbiome, with the relative abundance of potential probiotics such as Lactobacillus_johnsonii and Lactobacillus_taiwanensis been enriched.CONCLUSION:NPSs in SJZD can improve SDS-induced gastrointestinal-nervous system dysfunction through regulating microbiota-gut-metabolites axis, with four herbs exerting synergistic effects, which indicated the compatibility rationality of SJZD.

BACKGROUND:In China, Tongguanteng injection (TGT) is widely used in the treatment or adjuvant treatment of various types of cancer. However, the effect and mechanism of TGT in osteosarcoma is not clear. METHODS:The 143B and MG-63 cells were treated with different concentrations of TGT. Cell proliferation, migration, invasion and apoptosis were detected using CCK8 assay, transwell assay and flow cytometry. Differentially expressed genes (DEGs) were screened using RNA sequencing (RNA-seq). The identified mRNA and protein expression associated with the IRE1/CHOP pathway was validated by RT-PCR and western blot assay. To explore the underlying mechanisms, 4-phenylbutyric acid (4-PBA) was selected as a specific endoplasmic reticulum (ER) stress inhibitor. Small interfering RNA (siRNA) or pEX-3-ERN1 plasmid was transfected into 143B cells to silence or overexpress IRE1, respectively. The potential downstream proteins, including CHOP, and apoptosis associated proteins, caspase-3 and PARP1 were determined. Furthermore, the effect of TGT was demonstrated in 143B cell tumor-bearing mice in vivo. H&E staining, TUNEL staining and immunohistochemistry were conducted in tumor tissues obtained from the xenograft mouse model. RESULTS:TGT was shown to dramatically suppress the proliferation, migration and invasion, and induce apoptosis of osteosarcoma 143B and MG-63 cells in vitro. The identified DEGs included HSPA5 (encoding BiP) and ERN1 (encoding the IRE1 protein), as well as apoptosis-associated gene DDIT3 (encoding the CHOP protein). The term "IRE1-mediated unfolded protein response" was screened to be the most enriched biological process GO term. The expression of ER stress-associated proteins including ATF6, BiP, p-IRE1, XBP1s and CHOP, as well as apoptosis-associated cleaved caspase-3 and cleaved PARP1 proteins, was significantly upregulated by TGT treatment in osteosarcoma 143B cells, suggesting that TGT might promote the apoptosis of osteosarcoma 143B cells through the IRE1/CHOP pathway. Furthermore, knocking down IRE1 with si-IRE1 or inhibiting of ER stress with 4-PBA suppressed the expression of ATF6, BiP, XBP1s and CHOP induced by TGT, as well as the expression of cleaved caspase-3 and cleaved PARP1. On the contrary, overexpressing IRE1 promoted CHOP expression and induced osteosarcoma cell apoptosis. Consistent with in vitro results, TGT dramatically inhibited the tumor growth and promoted the expression of p-IRE1 and CHOP in tumor-bearing mice. CONCLUSION:The findings suggest that TGT exerts an anti-osteosarcoma effect in vitro and in vivo. The underlying mechanism might be associated with the activation of IRE1/CHOP pathway in ER stress. Our findings suggest that targeting IRE1/CHOP pathway might be a potential novel approach for osteosarcoma treatment.

ETHNOPHARMACOLOGICAL RELEVANCE:As a classical prescription for treating spleen deficiency syndrome (SDS), Sijunzi decoction (SJZD) is composed of Ginseng Radix et Rhizoma (RG, Panax ginseng C.A.Mey.), Atractylodes Macrocephalae Rhizoma (AM, Atractylodes macrocephala Koidz.), Poria (Poria cocos (Schw.) Wolf) and Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle (GRP, processed from Glycyrrhiza uralensis Fisch., Glycyrrhiza inflata Bat. or Glycyrrhiza glabra L.). The non-polysaccharides (NPSs) are the pharmacodynamic substance basis of SJZD, whose pharmacokinetics in SDS rats were elaborated previously. Further study on their tissue distribution and excretion properties is of significance for understanding the compatibility laws of SJZD.AIM OF THE STUDY:The aim was to unravel the tissue distribution and excretion characteristics of NPSs of SJZD in SDS rats, and explore the scientific connotation of SJZD compatibility.MATERIALS AND METHODS:A validated ultrafast liquid chromatography tandem mass spectrometry method was developed for monitoring the accurate dynamics of sixteen components in the tissues, feces and urine of SDS rats. The four incomplete formulae of SJZD were prepared by randomly deleting one herb to uncover the herb-herb interactions.RESULTS:All components of NPSs in SJZD were distributed in the tissues, except for ononin in the heart. Among them, glycyrrhetinic acid and atractylenolide III were more abundant in the liver and lung, respectively, while other components were enriched in the ileum, especially saponins. The evaluation of fecal excretion and urinary excretion revealed the low cumulative excretion of all components. The comparative analysis of incomplete formulae indicated that the tissue distribution and excretion became faster after removing Poria from SJZD, while a lack of RG led to slower tissue distribution. The tissue distribution at most time points was reduced when AM was absent. Further comprehensive visualization implied that SJZD compatibility can improve tissue distribution of the NPSs, especially ginsenosides and atractylenolide, at the specific time periods.CONCLUSION:The tissue distribution and excretion characteristics of NPSs of SJZD were elucidated in current research. Meanwhile, this study proposed new insights into the mechanism of SJZD compatibility rationality.

目的 研究通关藤Marsdenia tenacissima(Roxb.)Wight et Arn.的化学成分及其细胞毒活性.方法 通关藤水提取物采用大孔树脂、硅胶、ODS、制备HPLC进行分离纯化,根据理化性质及波谱数据鉴定所得化合物的结构.采用MTT法评价其对肿瘤细胞A549、Bel-7402的细胞毒活性.结果 从中分离得到12个化合物,分别鉴定为通关藤醇A(1)、12-O-惕各酰基-通关藤苷元 A(2)、isoshonanin(3)、caruilignan D(4)、香草酸(5)、丁香酸(6)、通关藤苷元A(7)、白桦脂酸(8)、11-O-异丁酰基-12-O-乙酰基通关藤苷元B-3-O-茯苓二糖苷(9)、通关藤苷H(10)、通关藤苷G(11)、通关藤苷Ⅰ(12).化合物1～2、7、10、12能抑制A549、Bel-7402细胞的生长.结论 化合物1为新化合物,化合物2为新天然产物,化合物3～6为首次从该植物中分离得到.化合物1～2、7、10、12具有细胞毒活性.

目的:研究木鳖子苷Ⅰ的化学结构及纯度测定方法.方法:采用2D NMR技术、水解方法确定化学结构及NMR数据归属;采用1H-NMR方法测定纯度.结果:运用2D NMR技术,结合酸水解、碱水解方法,快速确定了化学结构,首次完成了木鳖子苷Ⅰ NMR数据全归属.建立了1H-NMR方法测定木鳖子苷Ⅰ的纯度,该方法不需要特殊对照品,操作简单、快速准确.结论:本研究为复杂皂苷类成分的结构及纯度测定提供了方法借鉴.

目的 评价胃康灵胶囊中醇溶性多糖质量、生物活性.方法 分别采用比色法、高效凝胶渗透色谱-多角度激光散射-示差检测器联用(HPGPC-MALLS-RID)、HPLC、傅立叶红外变换光谱(FTIR)对多糖化学组成、重均相对分子质量、单糖组成、特征光谱进行分析,建立其HPLC、FTIR指纹图谱.采用微板法、CCK8法测定多糖对大肠杆菌的抑制作用及对人胃黏膜上皮细胞GES-1的保护作用.结果 12批样品醇溶性多糖中的中性多糖、糖醛酸、蛋白质平均含量分别为59.62％、12.53％、15.42％,重均相对分子质量、分布系数分别为4.19～18.57 kDa、1.007～1.154;含有甘露糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖,平均摩尔比为26.3:7.3:4.1:3.9:100.0:4.5:41.5;主要以吡喃糖形式存在.HPLC、FTIR指纹图谱相似度均大于0.90.醇溶性多糖对大肠杆菌具有抑制作用,对GES-1细胞具有保护作用,并呈剂量依赖性.结论 胃康灵胶囊中醇溶性多糖具有抑制大肠杆菌生长、保护GES-1细胞免受乙醇损伤的作用,其化学组成、重均相对分子质量及HPLC、FTIR指纹图谱有助于提升该制剂质量控制标准.

Osteosarcoma (OS) is a common primary malignant bone tumor in children and adolescents. The high recurrence and metastasis rate have become a common clinical problem to be solved, but there is no effective treatment. In recent years, studies have suggested that targeting the tumor microenvironment will likely become a new treatment direction for OS. Immune cell infiltration in the tumor microenvironment can promote tumor inflammation and angiogenesis. Tumor-associated macrophages (TAMs) are the most important immune cells in the tumor microenvironment, which play important roles in the development and metastasis of OS. The article reviews the effect of TAMs polarization on tumor cells and describes the effect of TAMs on the occurrence and development of OS from five aspects, including TAMs affecting the growth, invasion and metastasis, mediating chemotherapy resistance, stem cell-like phenotype, and immunosuppression of OS. The review summarizes the research progress of targeting TAMs in the treatment of OS in the past years, including influencing the recruitment of TAMs, promoting the polarization of M2 type to M1 type, targeting CD47 to promote the phagocytosis of TAMs, and targeting the immune checkpoint of TAMs, aiming to provide new directions and ideas for targeted therapy of OS.

Bao-Yuan Decoction (BYD), a widely used traditional Chinese medicine formula, is worth developing into modern dosage forms. To assess the quality of traditional decoction, the commonly used ultra-performance liquid chromatography coupled with diode array and evaporative light scattering detection (UPLC-DAD/ELSD) method was initially applied to develop the analytical methods for the qualitative fingerprints and simultaneous quantitation of multiple marker compounds in BYD. Based on 16 batches of BYD prepared from multiple batches of qualified crude herbs combined randomly, the characteristic fingerprints were generated, with 41 and 19 common peaks detected by DAD and ELSD, respectively. Furthermore, ginsenosides Re, Rg 1 and Rb 1 , calycosin-7-glucoside, calycosin, liquiritin, isoliquiritin apioside, isoliquiritin, glycyrrhizic acid and cinnamic acid were qualified as marker compounds to represent the herbs composing the formula. The characteristic fingerprints and the content ranges of multiple batches of the decoction were obtained, thus providing guidance for the quality control of modern dosage forms. The combination of these qualitative and quantitative methods will be an effective operational measure by which to evaluate and control the quality of BYD from traditional decoction to modern dosage forms.

Objective: To investigate the analgesic substances in the aerial part of Urtica fissa (Urticae Fissae Herba), commonly used for rheumatoid and rheumatism arthritis. Methods: The analgesic constituents were isolated with the active guidance of hot plate and acetic acid writhing models, and identified by comprehensive spectroscopic analysis. Results: Thirteen alkaloids (1-13), two lignans (14, 15), and three amides (16-18) were isolated from the active fractions. Among them, compound 1 was a new alkaloid, and compound 6 was a new natural product. The activity evaluation in vivo indicated that various pyrrole alkaloids (1, 3, 6, and 12) possessed significant analgesic activities, they could significantly inhibit the mice pain response induced by acetic acid and hot plate at the dosage of 2 mg/kg BW. Conclusion: The study revealed that the pyrrole alkaloids played important roles in the analgesic activities of Urticae Fissae Herba. (C) 2021 Tianjin Press of Chinese Herbal Medicines. Published by ELSEVIER B.V.

The chemical constituents in Urtica dioica fruits were investigated by silica gel chromatography, preparative HPLC, NMR, and HR-MS for the first time. As a result, 21 compounds were isolated from the fruits of U. dioica and identified 7R,8S,8'R-olivil(1), oleic acid(2), α-linoleic acid(3), palmic acid(4), methyl palmitate(5), α-linolenic acid(6), α-linolenic acid methyl ester(7), 5-O-caffeoyl-shikimic acid(8), vanillic acid(9), p-coumaric acid(10), 5-O-p-coumaroylshikimic acid(11), cinnamic acid(12), quinic acid(13), shikimic acid(14), ethyl caffeate(15), coniferyl ferulate(16), ferulic acid(17), caffeic acid(18), chlorogenic acid(19), pinoresinol(20), and quercetin(21). Compound 1 was a new compound and compounds 2-16 were isolated from U. dioica for the first time.

目的 优化急支糖浆现有薄层色谱鉴别和含量测定方法.方法 采用高效薄层色谱(HPTLC)法鉴别急支糖浆方中的四季青、鱼腥草、前胡、枳壳和甘草5种药材,采用高效液相色谱(HPLC)法测定原儿荼酸、盐酸麻黄碱和柚皮苷的含量.结果 HPTLC法结果显示,具栖冬青苷、白花前胡甲素、柚皮苷、甘草苷4种对照品和鱼腥草对照药材的薄层斑点均显示清晰,比移值均在0.2～0.8范围内;经验证,方法专属性、耐用性和批次重复性良好.HPLC法结果显示,原儿茶酸、盐酸麻黄碱和柚皮苷的线性范围分别为4.32～431.67、1.14～114.17、7.02～702.33μg/mL(r均大于0.996);平均加样回收率分别为100.61％、100.40％、99.22％ (RSD均小于2.00％);精密度(n=6)、稳定性(24h,n=7)、重复性(n=6)试验的RSD均小于2.00％.原儿茶酸等3种活性成分在10批急支糖浆中的平均含量依次为623.3、152.1、1213.9 μg/mL(RSD均小于10.00％).结论 本研究建立了急支糖浆一板多药味HPTLC鉴别方法,使用1种样品前处理方法、2个薄层色谱条件,实现了对处方中5种药材的快速鉴别;建立了HPLC含量测定方法,实现了对急支糖浆中3种活性成分的快速定量,可用于优化急支糖浆现有法定质量标准中的鉴别项和含量测定项.

Ethnopharmacological relevance: The root of Psammosilene tunicoides (W. C. Wu et C. Y. Wu) is a well-known medicinal herb for the treatment of pain, hemostasia and rheumatoid arthritis among Chinese people. Aim of the study: The present study aimed to investigate the antinociceptive activity and mechanism of beta-carboline alkaloids 1-4 which were extracted from the roots of P. tunicoides. Materials and methods: The analgesic effects were evaluated using peripheral and central pain mouse models of nociception, including the formalin test and the tail flick test. The levels of glutamic acid (Glu) and nitric oxide (NO) in cerebellar cortexes and spinal cords (L4-6) were determined. The anti-inflammatory of all compounds were then assessed on RAW264.7 cells. Results: Our results showed that compounds 1-4 had significant analgesic effects on both phases of formalin test of mice. Furthermore, all compounds showed suppressive effects on Glu in the brain and NO levels in the brain cortex and the spinal cord. Except for compound 1, the others could extend the pain threshold of hot water tail-flick in mice. In addition, compounds 2 and 3 (60 mu mol/kg) could decrease GABAA alpha 1 protein levels in spinal cord. All compounds exhibited anti-inflammatory effects by inhibiting lipopolysaccharide (LPS)-induced NO production in RAW264.7 cells with half-maximal inhibitory concentration (IC50) 1.1-34.9 mu M. Conclusion: beta-carboline alkaloids from the roots of P. tunicoides had significant analgesic activity by both central and peripheral mechanisms. Our findings suggested that regulating the release of NO or Glu or GABA alpha 1 are some of the mechanisms of analgesic activity of beta-carboline alkaloids.

Product photos are a very important part of product presentations in online shopping that directly aid consumers’ understanding of products. Previous studies have shown photo aesthetics matter to consumers. With the advances of image-processing and machine learning techniques, we are able to derive machine-based measures from photos and systematically examine their impact on e-commerce from multiple dimensions. In this study, we use a machine learning method to differentiate the foreground (i.e., product) and the background of a product photo and propose a background-composition-focus framework for understanding consumers’ perception of the photo. We conduct an empirical study using a real-world clothing dataset collected from one of the biggest fashion product websites in China. Using a hierarchical Bayesian model, we find that consumers prefer clothing products being shown on a simpler background, located vertically close to the center and horizontally following the rule-of-thirds. And, it is better to have a blurred background and no model face on the photo so that consumers can focus more on the product itself. These findings are of strong theoretical and practical value.

目的 研究急支糖浆多糖组分的化学成分组成、重均相对分子质量以及单糖组成,并比较不同批次急支糖浆多糖组分,为其质量控制提供依据.方法 采用比色法、高效凝胶色谱法(high performance gel permeation chromatography,HPGPC)和高效阴离子交换色谱法(high performance anion exchange chromatography,HPAEC)测定急支糖浆多糖的基本化学组成、重均相对分子质量及分散系数、单糖组成.结果 15批急支糖浆多糖的中性多糖、糖醛酸和蛋白质的平均质量分数分别为47.60％、33.38％、9.30％;重均相对分子质量及分散系数分别为11 757～26 367和1.89～2.65.15批急支糖浆多糖中均含有岩藻糖、鼠李糖、阿拉伯糖、半乳糖、葡萄糖、甘露糖、木糖、半乳糖醛酸及葡萄糖醛酸,平均质量分数分别为1.94、20.13、115.54、99.25、117.45、13.62、6.03、215.23、9.61 μg/mg,物质的量比结果为 1.5∶14.4∶100.0∶71.6∶84.7∶9.8∶5.2∶144.1∶6.4.将急支糖浆与川贝枇杷糖浆、杏苏止咳糖浆、小儿热速清糖浆和小儿肺热咳喘口服液进行比较,结果发现单糖组成物质的量比结合HPAEC指纹图谱可用于急支糖浆多糖的质量控制.结论 15批急支糖浆多糖的化学组成、重均相对分子质量及单糖组成具有较高的一致性,HPAEC指纹图谱具有特异性,可用作急支糖浆多糖的质控指标.

A quantitative analysis of multi-components by single marker(QAMS) method was established for the simultaneous determination of ephedrine hydrochloride, protocatechuic acid, 5-caffeoylquinic acid, 4-hydroxybenzoic acid, naringin, neohesperidin, glycyrrhizic acid, and praeruptorin A in Jizhi Syrup by high performance liquid chromatography(HPLC) with ultraviolet multi-wavelength detection system, and its feasibility in quality evaluation of Jizhi syrup was verified. With naringin as the internal reference substance, the relative correction factors and chromatographic peak localization methods of other seven components were respectively established at 210, 254, 280, and 320 nm. The method reproducibility was validated, and the result of QAMS were compared with those obtained by the external standard method(ESM) to verify the accuracy and feasibility of the method. The relative correction factors of ephedrine hydrochloride, protocatechuic acid, 5-caffeoylquinic acid, 4-hydroxybenzoic acid, neohesperidin, glycyrrhizic acid, and praeruptorin A with naringin as reference were 0.846, 0.582, 0.608, 0.293, 0.913, 2.207, and 0.940, respectively, which presented excellent reproducibility under different experimental conditions. Furthermore, QAMS and ESM showed no significant difference in the results for 15 batches of samples. Except protocatechuic acid and 5-caffeoylquinic acid, other six compounds were the exclusive components of single medicinal materials. In addition, glycyrrhizic acid and praeruptorin A were identified in the Jizhi Syrup for the first time, filling up the blank of no component detected in Glycyrrhizae Radix et Rhizoma and Peucedani Radix. The method established in this study is convenient, efficient, specific, accurate, and reliable, which can comprehensively and effectively evaluate the quality of Jizhi Syrup to ensure the safety and efficacy of this drug in clinical application.