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Chrysanthemum morifolium Ramat is an ornamental plant of worldwide cultivation. Like many other species in the family Asteraceae, C. morifolium is a rich producer of secondary metabolites. There are two objectives in this study: (I) to determine and compare the diversity of apolar secondary metabolites among different cultivars of C. morifolium and (II) to compare their properties as antifungal agents. To attain these objectives, we selected 13 cultivars of C. morifolium that are commonly used for making chrysanthemum tea as experimental materials. Leaves at the same developmental stage were collected from respective mature plants and subjected to organic extraction. The extracts were analyzed using gas chromatography-mass spectrometry. A total of 37 apolar secondary metabolites including 26 terpenoids were detected from the 13 cultivars. These 13 cultivars can be largely divided into three chemotypes based on chemical principal components analysis. Next, the extracts from the 13 cultivars were examined in in vitro assays for their antifungal properties against three species of pathogenic fungi: Fusarium oxysporum , Magnaporthe oryzae , and Verticillium dahliae . Significant variability in antifungal activity of the leaf extracts among different cultivars was observed. The 13 cultivars can be divided into four groups based on their antifungal activities, which could be partly correlated to the contents of terpenoids. In short, this study reveals large variations in chemical composition, particularly of terpenoids, of leaf secondary metabolites among different cultivars of C. morifolium and their different abilities in functioning as antifungal agents.

Carotenoids are the most important pigments determining the color of C. × morifolium; however, it is still unknown whether the changes of plastid ultrastructure affect carotenoids accumulation. In this study, we compared the change of carotenoid composition, content, and the plastid ultrastructures in the different developmental stages of capitulum among fourteen C. × morifolium cultivars from seven color groups. We found that the carotenoids and plastids detected at the early stage of capitulum development were similar in all cultivars, including violaxanthin, lutein, and β-carotene, which were present in proplastids and immature chloroplasts. Immature chloroplasts were degraded completely, forming loosely broken plastids during the development of the capitulum in white and pink cultivars. Meanwhile, a number of lipid vesicles appeared at proplastids, which resulted in only trace amounts of carotenoid accumulation in these cultivars. For yellow, orange, red, and brown cultivars, a great number of chromoplasts were found, which contained diverse ultrastructures, such as plastoglobules, tubules, and lipid droplets; these chromoplasts were derived from proplastids or chloroplasts. Compared with the early stage of capitulum development, these cultivars accumulated large amounts of carotenoids, primarily including lutein, lutein derivatives, and their isomers. In green cultivars, proplastids and immature chloroplasts were completely transformed into mature chloroplasts. These chloroplasts mainly contained violaxanthin, lutein, β-carotene, and two new components, (9Z)-lutein and (9'Z)-lutein, compared with carotenoid components presented in proplastids and immature chloroplasts. This research will be helpful for understanding the mechanisms of carotenoid metabolism of C. × morifolium. Furthermore, we found that two different chromoplast transformation patterns could be present in the same tissue cell, which contributed to the research on plastid differentiation and development in higher plants.

Background: Recent research has suggested that autophagy can provide a better mechanism for inducing cell death than current therapeutic strategies. This study investigated the effects of using an ethanol extract of Chrysanthemum zawadskii Herbich (ECZ) to induce apoptosis and autophagy associated with reliable signal pathways in mouse colon cancer CT-26 cells., Methods: Using ECZ on mouse colon cancer CT-26 cells, cell viability, annexin V/propidium iodide staining, acridine orange staining, reactive oxygen species (ROS) and western blotting were assayed., Results: ECZ exhibited cytotoxicity in CT-26 cells in a dose-dependent manner. ECZ induced apoptosis was confirmed by caspase-3 activation, poly (ADP-ribose) polymerase cleavage, and increased production of reactive oxygen species (ROS). Furthermore, it was shown that ECZ induced autophagy via the increased conversion of microtubule-associated protein 1 light chain 3II, the degradation of p62, and the formation of acidic vesicular organelles. The inhibition of ROS production by N-Acetyl-L-cysteine resulted in reduced ECZ-induced apoptosis and autophagy. Furthermore, the inhibition of autophagy by 3-methyladenine resulted in enhanced ECZ-induced apoptosis via increased ROS generation., Conclusion: These findings confirmed that ECZ induced ROS-mediated autophagy and apoptosis in colon cancer cells. Therefore, ECZ may serve as a novel potential chemotherapeutic candidate for colon cancer.

Chrysanthemum boreale is a plant widespread in East Asia, used in folk medicine to treat various disorders, such as pneumonia, colitis, stomatitis, and carbuncle. Whether the essential oil from C. boreale (ECB) and its active constituents have anti-proliferative activities in lung cancer is unknown. Therefore, we investigated the cytotoxic effects of ECB in A549 and NCI-H358 human lung cancer cells. Culture of A549 and NCI-H358 cells with ECB induced apoptotic cell death, as revealed by an increase in annexin V staining. ECB treatment reduced mitochondrial membrane potential (MMP), disrupted the balance between pro-apoptotic and anti-apoptotic Bcl-2 proteins, and activated caspase-8, -9, and -3, as assessed by western blot analysis. Interestingly, pretreatment with a broad-spectrum caspase inhibitor (z-VAD-fmk) significantly attenuated ECB-induced apoptosis. Furthermore, gas chromatography-mass spectrometry (GC/MS) analysis of ECB identified six compounds. Among them, β-caryophyllene exhibited a potent anti-proliferative effect, and thus was identified as the major active compound. β- Caryophyllene induced G 1 cell cycle arrest by downregulating cyclin D1, cyclin E, cyclin-dependent protein kinase (CDK) -2, -4, and -6, and RB phosphorylation, and by upregulating p21 CIP1/WAF1 and p27 KIP1 . These results indicate that β-caryophyllene exerts cytotoxic activity in lung cancer cells through induction of cell cycle arrest.

Chrysanthemum morifolium . cv "Hangju" is an important medicinal material with many functions in China. Flavonoids as the main secondary metabolites are a major class of medicinal components in "Hangju" and its composition and content can change significantly after flooding. This study mimicked the flooding stress of "Hangju" during flower bud differentiation and detected its metabolites in different growth stages. From widely targeted metabolomics data, 661 metabolites were detected, of which 46 differential metabolites exist simultaneously in the different growth stages of "Hangju". The top three types of the 46 differential metabolites were flavone C-glycosides, flavonol and flavone. Our results demonstrated that the accumulation of flavonoids in different growth stages of "Hangju" was different; however, quercetin, eriodictyol and most of the flavone C-glycosides were significantly enhanced in the two stages after flooding stress. The expression of key enzyme genes in the flavonoid synthesis pathway were determined using RT-qPCR, which verified the consistency of the expression levels of CHI , F3H , DFR and ANS with the content of the corresponding flavonoids. A regulatory network of flavonoid biosynthesis was established to illustrate that flooding stress can change the accumulation of flavonoids by affecting the expression of the corresponding key enzymes in the flavonoid synthesis pathway.

As an important "food and drug dual-use" product, chrysanthemums are widely used in both botanical medicine and food applications. However, the misuse of pesticides during chrysanthemum cultivation makes pesticide residue monitoring crucial. The aim of the present work was to address this practical demand for the simultaneous determination of multiple pesticide residues in various species of chrysanthemums. Both the sample pre-treatment and instrumental methods were systematically investigated. Seven chrysanthemum samples were extracted using acetonitrile and purified by dispersive solid-phase extraction with amino-modified multi-walled carbon nanotubes (MWCNTs-NH 2 ) and C18 as the cleanup co-adsorbents. After optimizing the amounts of MWCNTs-NH 2 and C18, matrix effects could not be avoided during LC-MS/MS analysis of 112 pesticides, although satisfactory recoveries were obtained. The use of SFC-MS/MS was evaluated, which demonstrated the significant positive role of SFC-MS/MS in reducing the matrix effects during pesticide residue analysis. In addition, the use of SFC-MS/MS permitted a shorter run time and afforded greater analytical efficiency. Method validation was further performed to evaluate the linearity, sensitivity, recovery, and precision of the developed method. Good linearity was observed for 92% of the analytes in the concentration range of 2-250 μg L -1 for all seven of the chrysanthemum samples. The LODs of the 112 pesticides ranged from 0.01 to 31.41 μg L -1 , depending on the sample, while the mean recoveries of all of the spiked pesticides ranged from 81.8% to 102% for concentrations of 20, 50, and 200 μg kg -1 . These results clearly demonstrate the applicability of the developed method for the simultaneous determination of multi-pesticides in various chrysanthemum samples., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Flos Chrysanthemi indici , an important medicinal and aromatic plant in China, is considered to have many different preservative and pharmacological properties. Considering the capability of essential oils (EOs), the present study is conducted to compare different extraction methods in order to improve yield and biological activities. Hydro-distillation (HD), steam-distillation (SD), solvent-free microwave extraction (SFME), and supercritical fluid extraction (SFE) are employed to prepare EOs from Flos Chrysanthemi indici . A total of 71 compounds are assigned by gas chromatography/mass spectrometry (GC-MS) in comparison with retention indices. These include 32 (HD), 16 (SD), 31 (SFME) and 38 (SFE) compounds. Major constituents of EOs differ according to the extraction methods were heptenol, tricosane, camphor, borneol, and eucalyptol. EOs extracted by SFME exhibit higher antioxidant activity. All EOs show varying degrees of antimicrobial activity, with minimum inhibitory concentration (MIC) ranging from 0.0625 to 0.125 mg/mL and SFME and SFE prove to be efficient extraction methods. EOs alter the hyphal morphology of Alternaria alternata , with visible bumps forming on the mycelium. Overall, these results indicate that the extraction method can significantly influence the composition and biological activity of EOs and SFME and SFE are outstanding methods to extract EOs with high yield and antimicrobial activity.

As dual regulators, the PTP-1B signaling pathway and α-glucosidase slow glucose release and increase the degree of insulin sensitivity, representing a promising therapeutic target for type 2 diabetes. In this study, we systematically examined the in vivo and in vitro anti-diabetic activities of natural flavonoids 1-6 from Chrysanthemum morifolium. Flavonoids 1-6 increased glucose consumption-promoting activity and the phosphorylation of GSK-3β and Akt, and decreased PTP-1B protein level along with slightly inhibitory activity of the PTP-1B enzyme. Moreover, flavonoids 1-2 treatment induced insulin secretion in INS-1 cells. Besides, in vivo study revealed that flavonoids 2 and 5 demonstrated potent anti-hyperglycemic and anti-hyperlipidemic activity, and improved maltose and glucose tolerance. Although flavonoid 2 exhibited lower inhibitory activity against α-glucosidase in vitro, it could deglycosylated in vivo to diosmetin to function as an α-glucosidase inhibitor. Taken together, these results led to the identification of the natural flavonoids 1-6 from C. morifolium as dual regulators of α-glucosidase and the PTP-1B signaling pathway, suggesting their potential application as new oral anti-diabetic drugs or functional food ingredients., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

In this study, phenolic profiles of chrysanthemums derived from five main species were determined for characterization of rationality of their application in tea, beverages and functional foods. A total of 41 phenolics including 3 phenolic acids, 17 flavones, 9 flavanones, 1 dihydroflavonol, 4 flavonols, 4 chalcones and 3 aurones were identified. The contents of 22 characteristic phenolics were simultaneously determined. Chrysanthemum morifolium cv. 'Qiju' (with abundant phenolics) and Coreopsis tinctoria Nutt. (with unique and abundant flavonoid aglycones and glycosides), exhibited excellent cellular antioxidant activities and strong market potentials. Chlorogenic acid, 3,5-dicaffeoylquinic acid and luteolin-7-O-glucoside largely contributing to cellular antioxidant activity of 'Qiju' by forming protective membrane around erythrocyte should be markers for quality control of 'Qiju'. Okanin, the gut microbial-produced metabolite of marein, possessed strong protective effect on oxidatively damaged erythrocyte via incorporating in membrane and entering cytoplasm. Okanin and marein should be markers for quality control of C. tinctoria., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The variation of flower color of chrysanthemum (Chrysanthemum×morifolium) is extremely rich, and carotenoids, which are mainly stored in the plastid, are important pigments that determine the color of chrysanthemum. However, the genetic regulation of the carotenoid metabolism pathway in this species still remains unclear. In this study, a pink chrysanthemum cultivar, 'Jianliuxiang Pink', and its three bud sport mutants (including white, yellow and red color mutants, 'Jianliuxiang White', 'Jianliuxiang Yellow' and 'Jianliuxiang Red', respectively) were used as experimental materials to analyze the dynamic changes of carotenoid components and plastid ultrastructure at different developmental stages of ray florets. We found that the carotenoid components and plastid ultrastructure of the four color cultivars in the early developmental stage of the chrysanthemum capitulum (S1) were almost identical, and the carotenoids mainly included violaxanthin, lutein and β-carotene, which exist in proplastids and immature chloroplasts. With the development of capitulum, the chloroplasts in 'Jianliuxiang White' and 'Jianliuxiang Pink' were degraded, and the protoplasts did not transform but rather formed vesicles that accumulated trace amounts of carotenoids. The proplastids and chloroplasts in 'Jianliuxiang Yellow' and 'Jianliuxiang Red' were all transformed into chromoplasts and consist of lutein as well as lutein's isomer and derivatives. Using comparative transcriptomics combined with gene expression analysis, we found that CmPg-1, CmPAP10, and CmPAP13, which were involved in chromoplast transformation, CmLCYE, which was involved in carotenoid biosynthesis, and CmCCD4a-2, which was involved in carotenoid degradation, were differentially expressed between four cultivars, and these key genes therefore should affect the accumulation of carotenoids in chrysanthemum. In addition, six transcription factors, CmMYB305, CmMYB29, CmRAD3, CmbZIP61, CmAGL24, CmNAC1, were screened using weighted gene co-expression correlation network analysis (WGCNA) combined with correlative analysis to determine whether they play an important role in carotenoid accumulation by regulating structural genes related to the carotenoid metabolism pathway and plastid development. This study analyzed dynamic changes of carotenoid components and plastid ultrastructure of the four bud mutation cultivars of chrysanthemum and identified structural genes and transcription factors that may be involved in carotenoid accumulation. The above results laid a solid foundation for further analysis of the regulatory mechanism of the carotenoid biosynthesis pathway in chrysanthemum., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

In this paper,the fingerprint of different varieties of chrysanthemum were established with " Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica" and the content of chlorogenic acid,galuteolin and 3,5-O-dicaffeoylquinic acid in 29 batches of different varieties of chrysanthemum in Futianhe town,Huangtugang town and Wuhan city were compared. At the same time,similarity evaluation and common peak clustering analysis were carried out. There were 11 common peaks in the fingerprints of 29 batches of different varieties of chrysanthemum,and the similarity ranged from 0. 802 to 0. 975. Hangju and Gongju were divided into one group by cluster analysis,and Huangju into another category. The established fingerprint method provides a basis for the identification of chrysanthemum cultivars. The content of 29 batches of chlorogenic acid was between 4. 092 and 11. 723 mg·g-1,luteolin was between 1. 010 and 11. 713 mg·g-1,and 3,5-O-dicaffeoylquinic acid was between 8. 828 and 33. 435 mg·g-1,both reach the pharmacopoeia standard,but the effective components of different varieties of chrysanthemum were quite different. Based on the contents of three active ingredients and the diversity of fingerprint peaks,the quality of the characteristic germplasm resource of local Fubaijuin Macheng is superior,and the protection of local characteristic germplasm resource should be strengthened in production.

The flowers of chrysanthemum species are used as a herbal tea and in traditional medicine. In addition, members of the genus have been selected to develop horticultural cultivars of diverse floral colors and capitulum forms. In this research, we investigated the phytochemical composition of eight gamma-irradiation mutant cultivars of Chrysanthemum morifolium and their original cultivars. The mutant chrysanthemum cultivars were generated by treatment with various doses of 60 Co gamma irradiation of stem cuttings of three commercial chrysanthemum cultivars as follows: 'ARTI-Dark Chocolate' (50Gy), 'ARTI-Purple Lady' (30 Gy), and 'ARTI-Yellow Star' (50 Gy) derived from 'Noble Wine'; 'ARTI-Red Star' (50 Gy) and 'ARTI-Rising Sun' (30 Gy) from 'Pinky'; 'ARTI-Purple' (40 Gy) and 'ARTI-Queen' (30 Gy) from 'Argus'; and 'ARTI-Rollypop' (70 Gy) from 'Plaisir d'amour'. Quantitative analysis of flavonoids, phenolic acids, anthocyanins, and carotenoids in the flowers of the 12 chrysanthemum cultivars was performed using high performance liquid chromatography-diode array detector-electrospray ionization mass spectrometry (HPLC-DAD-ESIMS). Essential oils from the flowers of these cultivars were analyzed by gas chromatography-mass spectrometry (GC-MS). The mutant cultivars, 'ARTI-Dark Chocolate', 'ARTI-Purple Lady', 'ARTI-Purple', and 'ARTI-Queen' showed higher total amounts of flavonoid and phenolic acid compared with those of the respective original cultivars. The mutant cultivars, 'ARTI-Dark Chocolate', 'ARTI-Purple Lady' and 'ARTI-Purple', which produce purple to pink petals, contained more than two-times higher amounts of anthocyanins compared with those of their original cultivars. Of the mutant cultivars, 'ARTI-Yellow Star' in which petal color was changed to yellow, showed the greatest accumulation of carotenoids. Ninety-nine volatile compounds were detected, of which hydrocarbons and terpenoids were abundant in all cultivars analyzed. This is the first report that demonstrated the phytochemical analysis of novel chrysanthemum cultivars derived from C. morifolium hydrid using HPLC-DAD-ESIMS and GC-MS. These findings suggest that the selected mutant chrysanthemum cultivars show potential as a functional source of phytochemicals associated with the abundance of health-beneficial components, as well as good source for horticulture and pigment industries.

Objective: To investigate the inhibitory effect of chrysanthemum extract on myocardial fibrosis in rats with renovascular hypertension, and explore the possible mechanism underlying this effect., Methods: Sixty Wistar rats were randomly divided into six groups: sham operation, model, positive control, and low-, medium-, and high-dose Huai chrysanthemum extract groups (ten rats per group). With the exception of the sham operation group, a renal hypertensive model was established in rats using the ""two-kidney, one clip"" method. After 6 weeks, low-, medium-, and high-dose groups were intragastrically administered chrysanthemum extract at 1, 2, or 4 g/kg, respectively, once daily for 4 weeks. The positive control group was administered Kato Pury at 50 mg/kg once daily for 4 weeks, while sham operation and model groups received an equal volume of distilled water once daily for 4 weeks. Blood pressure changes were examined before modeling, 6 weeks after modeling, and after 4 weeks of treatment administration. Ventricular remodeling indexes were measured by high frequency echocardiography after 4 weeks of treatment administration. Pathological changes were observed by hematoxylin and eosin, and Masson's trichrome staining methods. Collagen typeⅠ (Col Ⅰ) and type Ⅲ (Col Ⅲ) expression were examined by enzyme-linked immunosorbent assays. Transforming growth factor-β1 (TGF-β1), sma mad 3 (Smad3), Smad7, Ras homolog gene family, member A (RhoA), and Rho-associated protein kinase 1 (ROCK1) protein expression were detected by Western blot., Results: Compared with the model group, chrysanthemum-administered groups and the positive control group showed significant improvement of arterial blood pressure, echocardiography indicators, and degree of myocardial fibrosis (P < 0.05). In addition, these groups exhibited decreased expression of Col Ⅰ, Col Ⅲ, RhoA, ROCK1, TGF-β1, and Smad3, and increased Smad7 expression. Such improvements were most obvious in the high-dose chrysanthemum extract group (P < 0.05)., Conclusion: Chrysanthemum extract could effectively reduce myocardial fibrosis in rats with renovascular hypertension by a mechanism that potentially involves inhibition of RhoA/ROCK1 and TGF-β1/Smad signaling pathways.

One interesting phenomenon of Chrysanthemum morifolium tea is its formation of green or dark green color after hours of brewing. We investigated the greening reaction and its bioactivities, including an analysis of the green compounds. Results showed that the green color was due to a decrease in the L* (lightness), b* (yellowness/blueness), chroma values and an increase in hue angle. The green substances were found to be substances with similar polarities and unstable in acidic conditions. There was no significant difference (p < 0.01) in antioxidant activity between non-green and green samples. The green substances did not lead to cytotoxicity in PC12 cells at low concentrations, but at high concentrations, they caused a significant (p < 0.01) decrease in cell viability. The saccharide percentage and FT-IR results showed that the greening reaction was affected by the glycosides or groups attached to the saccharides, which might suggest a new mechanism for color-forming reactions., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Seventeen commercial chrysanthemum teas (Chrysanthemum morifolium and Coreopsis tinctoria) were extracted with hot-H 2 O, and examined and compared to the 75% methanol extracts for their chemical compositions using UPLC/Q-TOF-MS analysis. For the first time, 6, 8-C,C-diglucosylapigenin and eriodicyol-7-O-glucoside were detected in the Snow chrysanthemum, and acetylmarein was detected in HangJu, GongJu and HuaiJu. The extracts were also examined for their radical scavenging and anti-inflammatory activities in vitro. The hot-H 2 O extract of Kunlunmiju 1 had the greatest total phenolic content, and relative DPPH and oxygen radical absorbance capacity values of 12.72 mg gallic acid equivalents/g, 105.48 and 1222.50 μmol Trolox equivalents/g, respectively. In addition, all the hot-H 2 O extracts suppressed the lipopolysaccharide-induced interleukin-6, IL-1β and cyclooxygenase-2 mRNA expressions, and H 2 O 2 -induced intracellular reactive oxygen species production in cultured cells. The results from this research may be used to promote the consumption of chrysanthemum as a functional tea., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

This study aimed to elucidate the molecular mechanism of Chrysanthemum morifolium Ramat. against obesity and diabetes, by comparing the transcriptional changes in epididymal white adipose tissue (eWAT) with those of the bioactive compound in C. morifolium , luteolin (LU). Male C57BL/6J mice were fed a normal diet, high-fat diet (HFD), and HFD supplemented with 1.5% w/w chrysanthemum leaf ethanol extract (CLE) for 16 weeks. Supplementation with CLE and LU significantly decreased the body weight gain and eWAT weight by stimulating mRNA expressions for thermogenesis and energy expenditure in eWAT via lipid mobilization, which may be linked to the attenuation of dyslipidemia. Furthermore, CLE and LU increased uncoupling protein-1 protein expression in brown adipose tissue, leading to energy expenditure. Of note, CLE and LU supplements enhanced the balance between lipid storage and mobilization in white adipose tissue (WAT), in turn, inhibiting adipocyte inflammation and lipotoxicity of peripheral tissues. Moreover, CLE and LU attenuated hepatic steatosis by suppressing hepatic lipogenesis, thereby ameliorating insulin resistance and dyslipidemia. Our data suggest that CLE helps inhibit obesity and its comorbidities via the complex interplay between liver and WAT in diet-induced obese mice.

Background: Chrysanthemi Flos (CF), as a popular traditional Chinese medicine (TCM), has five main cultivars in China, namely "Chuju", "Boju", "Gongju", "Huaiju" and "Hangju". Due to their habitats and processing methods, great quality variations occur yet no systematical study has ever been carried out to evaluate such variations., Purpose: In this study, we aim to establish a new approach that can serve both as a quality control method and as an identification method for cultivars of CF., Method: The components in CF samples were identified by a combination of UPLC-ESI-Q-TOF/MS and GC/MS. Furthermore, a multimodal quantitative method was established by UPLC-UV coupled with principal component analysis (PCA) and the similarity evaluation system (SES), which was used to control and identify four cultivars of CF., Results: 18 compounds of flavonoids and caffeoylquinic acids were identified and ten of them were quantified using UPLC-ESI-Q-TOF/MS. Different cultivars of CF could be clearly distinguished with the fingerprints evaluation and principal component analysis (PCA). A total of 74 volatile compounds were detected by GC/MS. The distinctness of volatile components was observed. By the combination of UPLC-ESI-Q-TOF/MS and GC/MS, an identification and quality control method for CF was successfully established., Conclusion: The combination of UPLC-ESI-Q-TOF/MS and GC/MS could act as a comprehensive multimodal method for both identification and quality control of herbal medicines. This study provided new insights into the overall evaluation method for herbal medicines possessing different cultivars., (Copyright © 2018. Published by Elsevier GmbH.)

The capitula of Chrysanthemum morifolium and C. indicum are used to prepare Chrysanthemi Flos in traditional Japanese Kampo medicine. In our previous study, we reported on the agonistic effect of methanol extract of C. indicum capitulum on peroxisome proliferator-activated receptor (PPAR)-γ. We further isolated (E)-tonghaosu from C. indicum capitulum as one of the active ingredients. In the present study, we aimed to evaluate the PPAR-γ agonistic activity of a methanol extract of C. morifolium capitulum (MCM) in which (E)-tonghaosu could not be detected. MCM exhibited PPAR-γ agonistic activity in a concentration-dependent manner, and at a dose of 100 µg/ml, it showed similar activity to pioglitazone (30 µM), a standard PPAR-γ agonist. Through activity-guided fractionation, we isolated two geometric isomers, (E)- (1) and (Z)-B-ring-homo-tonghaosu (2), as the active ingredients of MCM. Both compounds exerted concentration-dependent PPAR-γ agonistic effects, and 1 had higher activity than 2. At 1.4 µM, 1 had similar activity to pioglitazone (30 µM), which was achieved by 2 at a concentration of 140 µM. Thus, 1 has the potential to become a lead compound for the drug discovery of PPAR-γ agonists. We compared the activities and the contents of (E)-, (Z)-tonghaosu, 1, and 2 among 13 commercial samples of Chrysanthemi Flos, including those derived from both C. morifolium and C. indicum. Their PPAR-γ agonistic activities were not related to the contents of these compounds. 1 and 2 were detected in the samples derived from both species but (E)- and (Z)-tonghaosu were not detected in the samples derived from C. morifolium; hence (E)- and (Z)-tonghaosu can serve as marker compounds to identify the capitula of C. indicum in Chrysanthemi Flos samples.

The feasibility of near-infrared spectroscopy (NIR) to detect chlorogenic acid, luteoloside and 3,5-O-dicaffeoylquinic acid in Chrysanthemum was investigated. An NIR spectroradiometer was applied for data acquisition. The reference values of chlorogenic acid, luteoloside, and 3,5-O-dicaffeoylquinic acid of the samples were determined by high-performance liquid chromatography (HPLC) and were used for model calibration. The results of six preprocessing methods were compared. To reduce input variables and collinearity problems, three methods for variable selection were compared, including successive projections algorithm (SPA), genetic algorithm-partial least squares regression (GA-PLS), and competitive adaptive reweighted sampling (CARS). The selected variables were employed as the inputs of partial least square (PLS), back propagation-artificial neural networks (BP-ANN), and extreme learning machine (ELM) models. The best performance was achieved by BP-ANN models based on variables selected by CARS for all three chemical constituents. The values of r p 2 (correlation coefficient of prediction) were 0.924, 0.927, 0.933, the values of RMSEP were 0.033, 0.018, 0.064 and the values of RPD were 3.667, 3.667, 2.891 for chlorogenic acid, luteoloside, and 3,5-O-dicaffeoylquinic acid, respectively. The results indicated that NIR spectroscopy combined with variables selection and multivariate calibration methods could be considered as a useful tool for rapid determination of chlorogenic acid, luteoloside, and 3,5-O-dicaffeoylquinic acid in Chrysanthemum.

Skin provides the first defense line against the environment while preserving physiological homeostasis. Subcutaneous tissues including fat depots that are important for maintaining skin structure and alleviating senescence are altered during aging. This study investigated whether theaflavin (TF) in green tea (GT) has skin rejuvenation effects. Specifically, we examined whether high ratio of TF contents can induce the subcutaneous adipogenesis supporting skin structure by modulating lipid metabolism. The co-fermented GT (CoF-GT) fraction containing a high level of TF was obtained by co-fermentation with garland chrysanthemum ( Chrysanthemum coronarium ) and the conventionally fermented GT (F-GT) fraction was also obtained. The effects of the CoF- or F-GT fractions on adipogenesis were assessed using primary human subcutaneous fat cells (hSCF). Adipogenesis was evaluated based on lipid droplet (LD) formation, as visualized by Oil Red O staining; by analyzing of adipogenesis-related factors by real-time quantitative polyperase chain reaction (RT-qPCR); and by measuring the concentration of adiponectin released into the culture medium by enzyme-linked immunosorbent assay. TF-enriched CoF-GT fraction did not adversely affect hSCF cell viability but induced their adipogenic differentiation, as evidenced by LD formation, upregulation of adipogenesis-related genes, and adiponectin secretion. TF and TF-enriched CoF-GT fraction promoted differentiation of hSCFs and can therefore be used as an ingredient in rejuvenating agents.

Ethnopharmacological Relevance: Chrysanthemum indicum Linne (C. indicum), a healthy food and folk medicine in China for thousands of years, has been reported to exert heat-clearing and detoxifying effects and extensively applied to treat various symptoms such as inflammation diseases, hepatitis and headache., Aim of This Study: The purpose of the present study was to investigate the protective effect of the supercritical carbon dioxide fluid extract from flowers and buds of C. indicum (CI SCFE ) on D-galactose-induced brain and liver damage during aging process and to illuminate the underlying mechanisms., Materials and Methods: Mice were orally administrated with CI SCFE (100, 150 and 300 mg/kg) after injection with D-galactose. 24 h after the last administration, the blood samples, whole brain and liver tissues were collected for biochemical analysis, histological examination and western blot analysis. The body weight, spleen and thymus indexes, alanine transaminase (ALT), aspartate transaminase (AST), total antioxidant capacity (T-AOC), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), malondialdehyde (MDA) in brain and liver, interleukin-1β (IL-1β), interleukin-6 (IL-6), and necrosis factor-α (TNF-α) were detected. Besides, the expressions of Bax, Bcl-2 and cleaved caspase-3 were determined by western blot assay., Results: The results indicated that CI SCFE effectively increased the suppressed body weight, attenuated the decline of thymus and spleen indexes, and reduced the elevated levels of ALT and AST induced by D-gal. Furthermore, CI SCFE might notably alleviate D-gal-induced abnormal alterations in structure and function of brain and liver dose-dependently via renewing normal antioxidant enzymes activities (SOD, CAT, GSH-Px), reducing MDA accumulation, decreasing inflammatory cytokines productions (IL-1β, IL-6, TNF-α), as well as attenuating the increase of Bax/Bcl-2 ratio and cleaved caspase-3 activation in the liver and brain., Conclusions: Taken together, our present results suggested that CI SCFE treatment could effectively mitigate the D-gal-induced hepatic and cerebral injury, and the underlying mechanism might be tightly related to the decreased oxidative stress, inflammation and apoptosis, indicating CI SCFE might be an alternative and promising agent for the treatment of aging and age-associated brain and liver diseases., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Dendranthema morifolium cv. 'jinsidaju', cultivated only in Kaifeng, has been eaten for more than 1000 years. During the antioxidant-activity-guided studies on its chemistry and health care function, two new bisabolane-type sesquiterpenes, (6R,7R)-7-hydroxybisabol-2,9E,11-triene-4-one (jinsidajuol A, 1) and (6R,7R)-7-hydroxy-11-methoxybisabol-2,9E-diene-4-one (jinsidajuol B, 2), and thirteen known compounds (3-15) were isolated from the flowers. Their structures were elucidated by 1D and 2D NMR spectroscopy and HRMS. 1 and 2 are the first example of bisabolane-type sesquiterpenes isolated from the genus Dendranthema. Compounds 6-8, 12 and 13 exhibited strong scavenging activities on the ABTS radical cation with IC 50 3.33, 5.67, 2.00, 2.50, 5.33 μg/mL, respectively. The IC 50 values of all compounds on HepG2 human hepatoma tumor cell line were higher than 50 μM., (Copyright © 2019 Elsevier B.V. All rights reserved.)

We previously reported that Chrysanthemum indicum polysaccharide (CIPS) effectively inhibited the replication of duck hepatitis A virus (DHAV). However, the inhibition mechanisms are still unclear. Autophagy plays important role in virus genomic replication. Therefore, in present study, the effect of autophagy on DHAV genome replication as well as the influence of CIPS on autophagy were studied. qPCR, western blot, and ELISA methods were applied to observe the autophagy and analyze the inhibition mechanisms of CIPS on DHAV. Results showed that DHAV infection increased the expression level of LC3-II and interdicted the degradation of p62. Treating with rapamycin benefited DHAV gene expression level. What's more, DHAV infection and rapamycin treatment also promoted the expression of PI3KC3 and increased the concentration of PI3P. However, CIPS treatment significantly downregulated the expressions of LC3-II and PI3KC3 induced by DHAV and rapamycin, and consequently inhibited autophagosomes formation. As a result, DHAV replication was inhibited., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

This study was aimed to develop novel antibacterial packaging materials in order to reduce the microbial contamination of food surface. Chrysanthemum essential oil (CHEO) was successfully incorporated into chitosan nanofibers (CS/NF) through electrospinning which was demonstrated by SEM and AFM analysis. The antibacterial mechanism of CHEO against Listeria monocytogenes (L. monocytogenes) was explored as well. The cell membrane permeability of L. monocytogenes appeared to be increased by CHEO. In addition, respiratory metabolism of L. monocytogenes was inhibited by CHEO through the inhibition of the Embden-Meyerhof-Parnas (EMP) pathway. The presence of CHEO had a negative effect on the activity of hexokinase, phosphofructokinase and pyruvate kinase in L. monocytogenes cells. Release efficiency study indicated that the CHEO could be released slowly from CHEO/CS/NF to achieve long-lasting antibacterial effect. The antibacterial application of the CHEO nanofibers against L. monocytogenes was tested on beef, with an inhibition rate of 99.91%, 99.97%, and 99.95% at the temperature of 4 °C, 12 °C and 25 °C, respectively, after 7 days of storage. Beef parameters like thiobarbituric acid reactive substances (TBARS), pH values, and texture at different storage temperatures (4 °C, 12 °C and 25 °C) were evaluated as well. Due to the presence of antioxidant components in CHEO released from CHEO/CS/NF, the TBARS value in treated beef was 0.135 MDA/kg lower (P < 0.05) than the untreated sample at 4 °C after 12 days. PH value assay indicated that PH value of beef sample packed with CHEO/CS/NF (6.43) was lower than unpacked sample (7.05) at 4 °C after 10 days of storage. These obtained results all illustrated the fact that CHEO/CS/NF could prolong the shelf-life of beef, suggesting a potential application in food packaging., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The Chrysanthemum morifolium flower is widely used in China and Japan as a food, beverage, and medicine for many diseases. In our work, two new caffeoylquinic acid derivatives ( 1, 2 ), a new flavanone glycoside ( 3 ), and six reported flavanones ( 4 ⁻ 9 ) were isolated and identified from the flowers of C. morifolium . The chemical structures of all isolates were elucidated by the analysis of comprehensive spectroscopic data as well as by comparison with previously reported data. The isolated constituents 1 ⁻ 8 were evaluated for their neuroprotective activity, and compounds 3 and 4 displayed neuroprotective effects against hydrogen peroxide-induced neurotoxicity in human neuroblastoma SH-SY5Y cells.

Twelve new guaianolide-type sesquiterpenoids (1-12) and five known guaianolide derivatives (13-17) were isolated from an aqueous ethanol extract of the aerial parts of Chrysanthemum indicum. Their structures were determined through spectroscopic data analysis. The absolute configurations of the new compounds were assigned by X-ray crystallography and electronic circular dichroism. Compound 5 shows multiple cytotoxic activities against four human naso-pharyngeal carcinoma (NPC) cell lines (CNE1, CNE2, SUNE-1, and HONE-1) and one human intestinal epithelial cell line (HT-29) with IC 50 values of 4.6, 6.0, 3.5, 4.3, and 9.6 μM, respectively. Compound 16 exhibits weak cytotoxicity against four NPC cell lines, CNE1 (IC 50 = 7.3 μM), CNE2 (IC 50 = 7.4 μM), HONE-1 (IC 50 = 7.6 μM), and SUNE-1 (IC 50 = 5.6 μM), but no cytotoxicity against HT-29 (IC 50 > 10 μM).

The aim of the present study was to explore whether the supercritical-carbon dioxide fluid extract from flowers and buds of Chrysanthemum indicum (SEC) exhibits antidepressant-like effects in a chronic unpredictable mild stress (CUMS)-induced mice model. Firstly, SEC was found to reverse a CUMS-induced decrease in the body weight gain in mice. Next, SEC was found to alleviate CUMS-induced depressive-like behavior, evidenced by the reversal of the decrease in the sucrose consumption in the sucrose preference test (SPT), the increase in the locomotor activity in the open field test (OPF), and the alleviation of immobility duration in both the forced swimming test (FST), and tail-suspension test (TST). SEC also attenuated CUMS-induced hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis by decreasing the levels of serum corticosterone and (CORT) and adrenocorticotropic hormone (ACTH), and hypothalamus corticotrophin-releasing hormone (CRH). In addition, SEC was found to suppress the expression of pro-inflammatory cytokines, including the tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 in the hippocampal of CUMS mice. Interestingly, further investigations demonstrated that SEC inhibited CUMS-induced activation of the nuclear factor kappa B (NF-κB) and NOD-like receptor protein 3 (NLRP3) inflammasomes pathways but upregulated brain-derived neurotrophic factor (BDNF) expression and promoted phosphorylation of extracellular signal-regulated kinase (ERK) and cAMP-response element-binding protein (CREB) in hippocampal. In summary, SEC was able to alleviate depressive-like behavior in a CUMS-induced mice model, accompanied by inhibitory roles in the hyperactivity of the HPA axis and pro-inflammatory cytokine expression. Modulating the NF-κB/NLRP3 and BDNF/CREB/ERK pathways contributed to SEC-mediated antidepressant-like effects.

Sunburn causes inflammation, which increases melanin production in skin and causes hyperpigmentation. Angiopoietin-like protein (ANGPTL) 2 is an inflammatory mediator induced in sun-exposed skin areas. However, whether ANGPTL2 functions in melanin production remains unclear. To assess this possibility, we overexpressed Angptl2 in the melanoma line B16 and in the keratinocyte line HaCaT. Relative to controls, Angptl2-expressing B16 cells produced higher melanin levels via tyrosinase induction. Accordingly, Angptl2-expressing HaCaT cells secreted relatively high levels of both endothelin-1 (ET-1) and α-melanocyte-stimulating hormone (α-MSH). Moreover, treatment with an extract from Chrysanthemum indicum × Erigeron annuus (CE) suppressed ANGPTL2 expression and repressed tyrosinase induction in melanocytes and of α-MSH and ET-1 in keratinocytes. Our data suggest that ANGPTL2 expression in keratinocytes and melanin-producing cells accelerates pigment production and that treatment of skin with a CE extract could prevent melanin accumulation., (© 2018 The Authors. Experimental Dermatology Published by John Wiley & Sons Ltd.)

Dietary chrysanthemum flower and wolfberry alone or together are widely consumed as a health beverage on a daily basis for centuries. The study aims to evaluate combinative effects of flower heads of Chrysanthemum morifolium cv. Hangju (C) and Lycium barbarum fruit (wolfberry, W) served as tea on chemical compounds, antioxidant and anti-inflammatory activities in RAW 264.7 macrophages. Eight phenolics were mainly detected in chrysanthemum flowers, whereas polysaccharides were dominant in wolfberry. The infusion of five combinations showed significantly antioxidant activities positively associated with the chrysanthemum flower content in chemical methods (ORAC and FRAP). However, the cellular-based CAA assay exhibited the highest antioxidant activities of the infusion at C:W = 1:1, indicating a synergistic interaction (CI = 0.11, P < .01). Additionally, the anti-inflammatory effect of infusion, specifically at a combination of C:W = 1:1, was observed by reducing the LPS-induced nitric oxide production, and inhibiting the expression of iNOS, TNF-α, IL-1β, and IL-6 mRNA (P < .05). The infusion prepared at a C:W = 1:1 was found to inactivate MAPKs (ERK and JNK) and NF-κB. The antioxidant and anti-inflammatory mechanisms might be attributed to acacetin-7-O-rutinoside, luteolin-7-O-glucoside and chlorogenic acid from chrysanthemum flower, and wolfberry polysaccharide via multiple inflammatory pathways., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Flower of Chrysanthemum morifolium is widely used in China and Japan as a folk medicine in treatment of many diseases. However, its active compounds remain largely unknown. In the present work, we have isolated, purified and characterized six new compounds (1-6), including two new arylnaphthalene lignans and four new phenolic glycosides, together with eight known compounds (7-14), from the flower of C. morifolium. Their structures and absolute configurations were elucidated in detail using 1D and 2D NMR, UV, IR, ORD, HRESIMS and ECD spectrometric data. In addition, compounds 1-3 possessed the significant neuroprotective activity against hydrogen peroxide-induced neurotoxicity in human neuroblastoma SH-SY5Y cells., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Enzyme treatment of the foods and herbs has been used to improve the absorption rate the efficiency of plant extracts by converting the glycosides of the plant into aglycones. In this study, we examined the obesity-inhibitory effect of Chrysanthemum indicum Linné (CI) treated with enzymes such as viscozyme and tannase, which are highly efficient in converting glycosides to aglycones and then compared with untreated CI extract. The enzyme-treated CI ethanol extract (CIVT) was administered orally at various doses for 7 weeks in the high fat diet (HFD)-fed male mice. CIVT administration reduced the body weights, the food efficiency and the serum levels of lipid metabolism-related biomarkers, such as triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-c) and leptin in the dose-dependent manner but not those high-density lipoprotein cholesterol (HDL-c) and adiponectin. CIVT also reduced considerably the total lipid amount in the liver and the size of adipocytes in the epididymal white adipose tissue (eWAT). CIVT effectively downregulated the adipogenesis-related transcription factors such as peroxisome proliferation activated receptor (PPAR)-γ and CCAAT/enhancer binding protein-α (C/EBP-α) but up-regulated PPAR-α, in the liver and eWAT. In addition, when compared to the enzyme-untreated CI 50% ethanol extract (CIEE), CIVT enhanced the reduction of body weight and lipid accumulation. Moreover, the viscozyme and tannase treatment of CI increased the flavonoid contents of the aglycone form. Therefore, our results support that the enzymatic treatment induced the production of aglycones for potentially suppressing the adipogenesis and lipid accumulation in HFD-fed mice. It suggests that CIVT might be an effective candidate for attenuating the over-weight and its related diseases.

Chrysanthemum morifolium Ramat is a perennial flowering plant widely cultivated for use in a tea infusion and as a popular beverage. To identify and evaluate the tea infusion made with a γ-irradiated mutant chrysanthemum cultivar with dark purple petals (cv. ARTI-Dark Chocolate), its phytochemical composition and antioxidant activity were tested and compared with those of the commercially available chrysanthemum cultivar with yellow petals (cv. Gamguk) by HPLC-DAD-ESIMS, as well as DPPH and ABTS radical scavenging assays. The purple chrysanthemum tea contained anthocyanins and linarin, which were not detected in the yellow chrysanthemum tea and the content of chlorogenic acid, acacetin-7- O -β-glucoside, and luteolin was higher compared with the yellow chrysanthemum tea. In contrast, the yellow chrysanthemum tea had higher luteolin-7- O -β-glucoside, 3,5-dicaffeoylquinic acid, apigenin-7- O -β-glucoside, and apigenin contents in comparison with the purple chrysanthemum tea. In addition, the content and antioxidant activity of the two chrysanthemum teas were investigated according to different water temperatures and infusing time. The yellow chrysanthemum tea did not show any significant differences according to infusing time and temperature, while the purple chrysanthemum tea was more influenced by the infusing time than water temperature, showing the highest total compound content in the infusing condition of 100 °C and 4 min. Moreover, the floral scent volatiles of the two chrysanthemum tea sources were analyzed using HS-SPME-GC-MS. In the DPPH radical scavenging assay, the purple chrysanthemum tea broadly showed greater antioxidant activity than did the yellow chrysanthemum tea, corresponding to the high content of anthocyanins known as the powerful antioxidant. Further, both chrysanthemum flower teas exhibited strong ABTS radical scavenging effects ranging from 76% to 61% under all infusing conditions. Therefore, the purple chrysanthemum cultivar, ARTI-Dark Chocolate, is worthy of breeding as a new tea cultivar.

Tea brewed from chrysanthemum flowers has been widely used in Chinese medicine. The possibly medicinal compounds in Chrysanthemum morifolium tea can be purified by preparative high performance liquid chromatography (HPLC), but this is usually done with acidic conditions, which leads to the hydrolysis of glycosides. In hopes of avoiding this hydrolysis, we explored the effect of weakly basic conditions on the separation and purification of flavonoids and glycosides from Chrysanthemum morifolium . We also explored the effects of weakly basic conditions on chlorogenic acid (3-CQA) and apigenin-7- O -glucoside (A7G). Our results show that the concentration of the weakly basic ammonium hydrogen carbonate and time had no significant effect on A7G, p < 0.01, but it had a significant effect on 3-CQA, p < 0.01. HPLC and ultraviolet (UV) analysis showed that the structure of 3-CQA is destroyed in weakly basic conditions. Caffeic acid, quinic acid, and 3,4-dihydroxymandelic acid, which is a hydrolysate of 3-CQA, were identified by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The results showed that weakly basic conditions could be used for the purification of flavonoids and glycosides but not for caffeoylquinic acids. Moreover, our work clarified the hydrolysis behaviour of caffeoylquinic acids, which can be helpful for research into their functional aspects.

Obesity is directly associated with cancer, cardiovascular injury, hypertension, and type 2 diabetes. To date, Yamamoto identified that hot water extracts of edible Chrysanthemum (EC) induced cell size reduction, up-regulation of adiponectin expression, and glucose absorption inhibition in 3T3-L1 cells during adipocyte differentiation. Furthermore, EC showed antidiabetic effects such as improvement in insulin resistance and the down-regulation of the blood glucose level and liver lipid content in type 2 diabetes model mice. In this study, we attempted to identify the antidiabetic components in EC. The methanol fraction from EC that showed relatively strong biological activity was purified by chromatography to obtain acacetin-7-O-glucoside, apigenin-7-O-glucoside, kaempferol-7-O-glucoside, and naringenin-7-O-glucoside. Among the isolated compounds and their aglycones, naringenin (NA) and naringenin-7-O-glucoside (NAG) up-regulated the intracellular accumulation of lipid and adiponectin-secretion and down-regulated the diameter of 3T3-L1 cells during adipocyte differentiation. Because the PPARγ antagonist BADGE and PI3K/Akt inhibitors wortmannin and LY29004 inhibited the intracellular lipid accumulation by NA and NAG associated with adipogenesis, it was considered that NA and NAG showed the above-mentioned activities via the activation of PPARγ as well as phosphorylation of the PI3K/Akt pathway., (© 2019 Wiley-VHCA AG, Zurich, Switzerland.)

The effect of Chrysanthemum morifolium flower extract (CME) was investigated on the formation of heterocyclic amines (HCAs) in goat meat patties cooked at different temperatures by different cooking methods. In patties without CME, the level of total HCAs increased analogously with the increase in cooking temperature. The inhibitory effect of CME on HCAs ranged from 14% to 82%. The total HCAs content were reduced by 46%, 40% and 35% in pan fried, deep fat fried and oven roasted goat meat patties, respectively, at 175 °C. While, at 225 °C, the amount of total HCAs was decreased by 52%, 47% and 32% in deep fat fried, pan fried and oven roasted patties respectively, whereas, in barbecue patties the reduction was 36%. Hence, it is concluded that the effect of CME was more prominent in deep fat frying and pan frying cooking methods and increase in temperature enhance the efficacy of CME in both cooking methods., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Apigenin-7-O-glucoside is an active phenolic compound in Asteraceae flowers and possesses remarkable therapeutic applications. However, its high price and low abundance in plants limit its use, meanwhile it would hydrolyze in the purification process. In this study, apigenin-7-O-glucoside extracted with ultrasound and purified with preparative HPLC from Chrysanthemum morifolium 'Huangju' was investigated, as well as its hydrolysis behavior and bioactivities. The optimized extraction conditions were: solid/liquid ratio: 1:20, extraction time: 35 min, temperature: 50 °C, and ultrasound power: 350 W. The content of apigenin-7-O-glucoside was up to 16.04 mg/g. Apigenin-7-O-glucoside was then purified with preparative HPLC from the extract, and confirmed by Q-TOF/MS. Apigenin-7-O-glucoside was partially hydrolyzed in acidic condition, and the hydrolysis rate depended on the pH value and temperature. The antioxidant activity increased as a result of the hydrolysis process. This study provided a green and effective way to obtain apigenin-7-O-glucoside and would be beneficial for further investigations into nutritional and functional aspects apigenin-7-O-glucoside and other glycosides.

The key point of our work was evaluating the impact of terpene alcohols on the aroma expression of terpenes recombination in Chrysanthemum essential oils. Using pure commercial products, various aromatic recombinations were prepared, consisting of terpenes recombination and six terpene alcohols, all the concentrations found in Chrysanthemum essential oils. There were five groups of terpene alcohols mixtures performed very interesting with the addition or omission tests. The "olfactory threshold" of the terpenes recombination had a notable decrease when adding isoborneol, d-Fenchyl alcohol respectively through the Feller's additive model analysis. Furthermore, the descriptive test indicated that the addition of terpene alcohols mixture had the different effect on fruity, floral, woody, green, and herbal aroma intensity. Specifically, when isoborneol was added to the terpenes recombination in squalane solution, it was revealed that isoborneol had a synergy impact on herbal and green notes of the terpenes recombination and masked the fruity note.

Chrysanthemum indicum Linne flower (CF) and Cinnamomum cassia (L.) J. Presl bark (CB) extracts have been used as the main ingredients in several prescriptions to treat the hyperuricemia and gout in traditional medicine. In the present study, we investigated the antihyperuricemic effects of DKB114, a CF, and CB mixture, and the underlying mechanisms in vitro and in vivo. DKB114 markedly reduced serum uric acid levels in normal rats and rats with PO-induced hyperuricemia, while increasing renal uric acid excretion. Furthermore, it inhibited the activity of xanthine oxidase (XOD) in vitro and in the liver in addition to reducing hepatic uric acid production. DKB114 decreased cellular uric acid uptake in oocytes and HEK293 cells expressing human urate transporter (hURAT)1 and decreased the protein expression levels of urate transporters, URAT1, and glucose transporter, GLUT9, associated with the reabsorption of uric acid in the kidney. DKB114 exerts antihyperuricemic effects and uricosuric effects, which are accompanied, partially, by a reduction in the production of uric acid and promotion of uric acid excretion via the inhibition of XOD activity and reabsorption of uric acid. Therefore, it may have potential as a treatment for hyperuricemia and gout.

The rapid and nondestructive determination of active compositions in Chrysanthemum morifolium (Hangbaiju) is of great value for producers and consumers. Hyperspectral imaging as a rapid and nondestructive technique was used to determine total polysaccharides and total flavonoids content in Chrysanthemum morifolium . Hyperspectral images of different sizes of Chrysanthemum morifolium flowers were acquired. Pixel-wise spectra within all samples were preprocessed by wavelet transform (WT) followed by standard normal variate (SNV). Partial least squares (PLS) and least squares-support vector machine (LS-SVM) were used to build prediction models using sample average spectra calculated by preprocessed pixel-wise spectra. The LS-SVM model performed better than the PLS models, with the determination of the coefficient of calibration (R²c) and prediction (R²p) being over 0.90 and the residual predictive deviation (RPD) being over 3 for total polysaccharides and total flavonoids content prediction. Prediction maps of total polysaccharides and total flavonoids content in Chrysanthemum morifolium flowers were successfully obtained by LS-SVM models, which exhibited the best performances. The overall results showed that hyperspectral imaging was a promising technique for the rapid and accurate determination of active ingredients in Chrysanthemum morifolium , indicating the great potential to develop an online system for the quality determination of Chrysanthemum morifolium .

This study identified the volatile organic compounds in the essential oils that are extracted from Chrysanthemum indicum Linné ( C. indicum Linné) and investigated the effects of the inhalation of these compounds. We detected a total of 41 volatile organic compounds, including 32 hydrocarbons, four acids, three alcohols, two ketones, and one aldehyde. In a sniffing test, seven types of volatile organic compounds were identified. Furthermore, the volatile organic compounds in C. indicum Linné that were identified were found to be derived from 1,8-cineole and camphor. After inhalation of the essential oils, the subjects' systolic blood pressure and heart rate decreased. This indicates that inhalation of the essential oils extracted from C. indicum Linné provides mental and physical relaxation. We examined the changes in electroencephalogram findings that are observed after C. indicum Linné essential oil inhalation. An increase in theta and alpha waves, which usually appear during relaxation, as well as a decrease in beta and gamma waves, which appear during brain activity such as excessive attention, were noted. These results indicate that C. indicum Linné essential oil inhalation helps to reduce blood pressure and may provide mental and physical relaxation.

Targeting the two degradation systems, ubiquitin proteasome pathway and ubiquitin signal autophagy lysosome system, plays an important function in cancer prevention. Borneol is called an "upper guiding drug". Luteolin has demonstrated anticancer activity. The fact that borneol regulates luteolin can be sufficient to serve as an alternative strategy. Borneol activates luteolin to inhibit E1 and 20S activity (IC 50 = 118.8 ± 15.7 μM) and perturb the 26S proteasome structure in vitro. Borneol regulates luteolin to inhibit 26S activity (IC 50 = 157 ± 19 μM), induces apoptosis (LC 50 = 134 ± 4 μM), and causes pre-G1 and G0/G1 arrest in HepG2 cells. Borneol regulates luteolin to induce ubiquitin signal autophagic degradation, resulting in induction of E1, reduction of USP47, and accumulation of p62 in HepG2 reporter cells. Interestingly, luteolin decreased Ub conjugates, while borneol increased the accumulation of Ub conjugates in HepG2 reporter cells. E1, p62, and ubiquitin levels were downregulated in borneol-treated HepG2 reporter cells at 24 h. These observations suggest a potential autophagic inhibitor of borneol that may guide luteolin in the ubiquitin proteasome pathway and the ubiquitin signal autophagic degradation.

A new glycoside, named as CG-1, was separated from Imperial Chrysanthemum with silica gel column chromatography. The purity was detected by TLC and HPLC. The crystal shape of CG-1 was consisting of a quadrangular and two rectangular pyramids. The analysis of DSC and TGA showed that the melting point of CG-1 crystal was 150.22 °C and had good thermal stability. The monosaccharide conformation analysis showed that it was d-glucose. The structure characteristics were compared by FT-IR, NMR spectroscopy and LC-MS and a molecular structure has been deduced which consistent with spectroscopic results. In vitro antioxidant results suggested that the glycoside extracted from Imperial Chrysanthemum could be effectively employed as natural antioxidant in health or functional food. This work is of significance to keep the antioxidant activity in the processing and application of Imperial Chrysanthemum., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

To investigate the preventive effect of Kunlun snow chrysanthemum polysaccharides (KSCP) on acetaminophen (AP) induced liver damage and its possible mechanism. Mice acute liver injury model was established via intraperitoneal injection of AP (300 mg/kg). The biochemical indicators of plasma and liver tissue were tested. The effects of KSCP on the liver index were examined. The liver pathological changes were investigated. The expressions of related protein were detected via Western blotting. In our study, compared with model group, the concentrations and contents of ALT, AST, TNF-α, IL-1β and MDA were reduced and activities of SOD were increase in H-KSCP (1.2mg/10 g)-pretreated mice (P<0.01). The liver index was significantly reduced in H-KSCP-pretreated mice compared with model group (4.89±0.22 vs 7.4±0.66, P<0.01). Liver cellular swelling, degeneration and necrosis relieved, and pathological injury had been improved. Western blotting results showed that the caspase-3 protein level in H-KSCP group was significantly decreased, expression of Bcl-2 protein and Bcl-2/Bax ratio was increased, whereas which of Bax protein was decreased (P<0.01). KSCP-pretreated at middle and high doses can prevent against the liver injury, its action mechanism may be related to its anti-inflammatory effects and regulation of apoptosis related proteins expression. Overall, our results showed that KSCP may be an effective preventive agent in preventing acute liver injury.

A fast, sensitive and reliable method for the determination of fipronil and its metabolites in tea and chrysanthemum was developed using a modified QuEChERS technique and an ultra performance liquid chromatography Q-Exactive Orbitrap mass spectrometry. The mixture of adsorbents containing primary secondary amine (PSA), octadecylsilane (C 18 ) and carbon nanotubes (CNTs), was used as QuEChERS adsorbents. The use of mass resolution at 70000 full width at half maximum (FWHM) and narrow mass windows at 5 ppm achieved high selectivity and repeatability. Satisfactory linearity with correlative coefficient (R 2 ) higher than 0.996 was achieved for all compounds. Recoveries at three levels (2, 10 and 50 μg kg -1 ) ranged from 86% to 112%, while the intra- and inter-day accuracies were less than 15%. Limits of quantification for fipronil and its metabolites were 2 μg kg -1 , which fulfils the requirement of maximum residue limits formulated by European Union and Japan., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Dendranthema indicum var. aromaticum , which is an aromatic plant with a strong and special fragrance throughout the whole plant, is used for the treatment of colds and headaches, and as a mosquito repellant in Shennongjia, Hubei province, China. To analyze the composition of the essential oil from this medicinal herb, we developed a gas chromatography-mass Spectrometry (GC-MS) method including microwave-assisted extraction, hydrodistillation and direct headspace analysis in two different stationary phase columns. In total, 115 volatile compounds were identified, of which 90 compounds were identified using Rxi-5MS and 78 using HP-INNOWAX. Our results revealed that the oil was mainly composed of five categories of compound: oxygenated monoterpenes (28.76-78.10%), oxygenated sesquiterpenes (4.27-38.06%), sesquiterpenes (3.22-11.57%), fatty hydrocarbons (1.65-9.81%) and monoterpenes (0-3.32%). The major constituents are α-thujone, β-thujone, cis -sabinol, sabinyl acetate and (-)-neointermedeol.However, the essential oil composition in the published literature differs significantly. Therefore, a cluster analysis was carried out using the top ten compositions in the reported literature as well as this study, using Minitab software. To provide detailed information on plant origin, the ITS1-5.8s-ITS2 region was amplified and sequenced (Accession No. MF668250). Besides, in order to provide a macroscopic view of the chemical composition, the biosynthetic pathway of the main components was summarized according to the Kyoto Encyclopedia of Genes and Genomes (KEGG) database and the published literatures., Competing Interests: The authors declare no conflict of interest.

Ten new highly oxidized monomeric (1-8) and dimeric guaianolides (9 and 10), along with two known guaianolide derivatives (11 and 12), were isolated from the aerial parts of Chrysanthemum indicum using a bioassay-guided fractionation procedure. The new compounds were characterized by the basic analysis of the spectroscopic data obtained, and the absolute configurations were determined by both empirical approaches and ECD calculations. Inhibitory effects of 1-12 on nitric oxide production were investigated in lipopolysaccaride (LPS)-mediated RAW 264.7 cells, and most of them (1-8 and 11) displayed IC 50 values in the range 1.4-9.7 μM. Moreover, a mechanistic study revealed that the potential anti-inflammatory activity of compound 1 appears to be mediated via suppression of an LPS-induced NF-κB pathway and down-regulation of MAPK activation.

In the present study, the chemical compositions and skin whitening-related antioxidant and anti-melanogenic effect of essential oils (EOs) extracted from Chrysanthemum borealeMakino (CBM) (CBMEOs) at vegetative, pre-flowering and full-flowering are investigated and contrasted among the three stages. The yields and components of the CBMEOs were different at each stage. The CBMEOs increased DPPH and ABTs scavenging activities and attenuated the α-melanocyte stimulating hormone (α-MSH)-induced tyrosinase activity and melanin synthesis in B16BL6 cells. Among CBMEO components, eugenol had the highest DPPH and ABTs scavenging activities and cuminaldehyde was the strongest inhibitor of α-MSH-induced tyrosinase activity and melanin synthesis. The CBMEOs in each stage showed the different levels of phosphorylation of extracellular signal-regulated kinase1/2 and p38 MAPK. These findings demonstrate that the CBMEOs have antioxidative and anti-melanogenic activities in all the CBM harvesting stages, resulting in skin-whitening biological activities and that the levels of their component contents and bioactivities differ among the CBM harvesting stages. The CBMEOs may have the potential for use in cosmetics and alternative medicine., (© 2018 Wiley-VHCA AG, Zurich, Switzerland.)

Chrysanthemum indicum (CI) is widely distributed in China and many parts of the tropical world, and has been reported to have antibacterial, antiviral, anti-oxidant and immunomodulatory effects, but no information is available on its effects on high fat diet (HFD)-induced obesity. This was undertaken to investigate the mechanism responsible for the effect of ethyl acetate fraction of CI (CIEA) on adipogenesis, in vitro and in vivo models of obesity. In the in vitro study, differentiating 3T3-L1 cells were treated with media to initiate differentiation (MDI) in the presence or absence of CIEA with different concentrations, and in the in vivo study, C57BL/6 mice were fed with HFD and administered CIEA daily for six weeks. Garcinia cambogia (GC) was used as the positive control, and was administered in the same manner as CIEA. Results showed CIEA reduced HFD-induced body weight gain, epididymal white adipose tissue (eWAT), and liver weight. In addition, CIEA significantly decreased serum lipid profiles, including total cholesterol (TC), triglyceride (TG) and low density lipoprotein cholesterol (LDLc) and increased high density lipoprotein cholesterol (HDLc) levels. Furthermore, CIEA also reduced leptin levels and increased adiponectin levels in serum, and significantly decreased peroxisome proliferator-activated receptor [Formula: see text] (PPAR[Formula: see text]) and CCAAT/enhancer-binding protein (C/EPBs) levels, but increased PPAR[Formula: see text] level and the phosphorylation of AMP-activated protein kinase (AMPK) in eWATs and in the liver tissues of HFD fed obese mice. Taken together, these results indicate CIEA might be beneficial for preventing obesity.