RRLC-QTOF/MS-Based Metabolomics Reveal the Mechanism of Chemical Variations and Transformations of Astragali Radix as a Result of the Roasting Process.

Astragali Radix (AR), which is extensively used as a healthy food supplement and medicinal herb, contains two forms of products corresponding to raw Astragalus Radix (RAR) and processed Astragali Radix (PAR), which was obtained by roasting. In this study, a non-targeted rapid resolution liquid chromatography coupled with quadruple time-of-flight mass spectrometry (RRLC-Q/TOF-MS) based metabolomics approach was developed to investigate the chemical changes of AR due to roasting. A total of 63 compounds were identified or tentatively identified. Among them, 23 isoflavonoids (composed of 12 isoflavones, eight pterocarpans, and three isoflavans) and six cycloastragenols were characterized as differential metabolites. Heatmap visualization and high-performance liquid chromatography coupled with photodiode array and evaporative light scattering detector (HPLC-PDA-ELSD) quantitative analysis revealed that malonyl isoflavonoids or cycloastragenols were at higher levels in RAR. These might be converted to corresponding acetyl isoflavonoids and cycloastragenols and related isoflavonoid glycosides during roasting. To prove this prediction, chemical conversion experiments on malonyl isoflavonoids and cycloastragenols were performed to confirm and clarify the chemical transformation mechanism.

Oxymatrine Inhibits Colorectal Cancer Metastasis via Attenuating PKM2-Mediated Aerobic Glycolysis.

BACKGROUND: Colorectal cancer (CRC), a type of highly occurred intestinal cancer at present, is prone to metastasis at the later stage of chemotherapy. Looking for the anti-metastatic agents from natural compounds attracted much concern. Here, it aims to demonstrate whether oxymatrine, an anti-cancer natural compound, has anti-metastatic activity and its potential significance in clinic. MATERIALS AND METHODS: Wound healing assay and transwell assay were for evaluating the effect of oxymatrine on cell migration and invasion in vitro. Anti-metastatic action in vivo was determined by hepatic metastasis of colorectal cancer cells in mice. RESULTS: Oxymatrine can significantly inhibit cancer cell migration and invasion in vitro. The production of ATP, pyruvate, and lactate was suppressed in CRC cells under the treatment of oxymatrine, as well as the glucose consumption. Meantime, extracellular acidification rates (ECR) were evidently attenuated although the oxygen consumption rates (OCR) were not affected. Both clued that oxymatrine inhibition of metastasis is possibly related to blocking aerobic glycolysis. Subsequent results indicated that pyruvate kinase M2 (PKM2) not hexokinase (HK) and phosphofructokinase (PFK) were involved in oxymatrine blocking glycolysis as the PKM2 kinase activity and expression were inhibited by oxymatrine and the PKM2 activator, TEPP-46, can reverse in part the effect of oxymatrine induced in CRC cells. Furthermore, this process was also mediated by inhibition of glucose transporter 1 (GLUT1). Finally, the in vivo metastatic model in mice showed both 20 mg/kg and 40 mg/kg oxymatrine significantly inhibit liver metastasis of CRC cells in mice, and PKM2 and GLUT1 expression in liver of the oxymatrine-treated group is declined. CONCLUSION: Oxymatrine exerted anti-metastatic activity dependent on inhibition of PKM2-mediated aerobic glycolysis. It is not only an anti-cancer agent but also a potential anti-metastatic compound with clinical application significance.

Human serum albumin-mediated recognition of soluble amyloid-ß peptides using a time-resolved luminescent probe in plasma.

A terbium(iii) complex can recognize soluble Aß in plasma through human serum albumin (HSA)-mediated co-assembly, which can not only circumvent the interference of HSA, but also benefit Aß enrichment with amplified time-resolved luminescence enhancement.

Structural Identification and Conversion Analysis of Malonyl Isoflavonoid Glycosides in Astragali Radix by HPLC Coupled with ESI-Q TOF/MS.

In this study, four malonyl isoflavonoid glycosides (MIGs), a type of isoflavonoid with poor structural stability, were efficiently isolated and purified from Astragali Radix by a medium pressure ODS C18 column chromatography. The structures of the four compounds were determined on the basis of NMR and literature analysis. Their major diagnostic fragment ions and fragmentation pathways were proposed in ESI/Q-TOF/MS positive mode. Using a target precursor ions scan, a total of 26 isoflavonoid compounds, including eleven malonyl isoflavonoid glycosides coupled with eight related isoflavonoid glycosides and seven aglycones were characterized from the methanolic extract of Astragali Radix. To clarify the relationship of MIGs and the ratio of transformation in Astragali Radix under different extraction conditions, two MIGs (calycosin-7-O-glycoside-6″-O-malonate and formononetin-7-O-glycoside-6″-O-malonate) coupled with related glycosides (calycosin-7-O-glycoside and formononetin-7-O-glycoside) and aglycones (calycosin and formononetin) were detected by a comprehensive HPLC-UV method. Results showed that MIGs could convert into related glycosides under elevated temperature conditions, which was further confirmed by the conversion experiment of MIGs reference compounds. Moreover, the total contents of MIGs and related glycosides displayed no obvious change during the long-duration extraction. These findings indicated that the quality of Astragali Radix could be evaluated efficiently and accurately by using the total content of MIGs and related glycosides as the quality index.

[Comparative study of water characteristic components of Glycyrrhiza uralensis from three geographical regions by chemical pattern recognition combined with muti-component qualitative and quantitative analysis].

Thirty-two batches of cultivated and wild Glycyrrhiza uralensis were obtained from three geographical regions. Comparative study of water characteristic components of G. uralensis from three geographical origins was conducted by PCA,OPLS-DA chemical pattern recognition combined with LC-TOF/MS and muti-component analysis. The similarity of fingerprints of 32 batches of medicinal materials ranged from 0. 903 to 0. 999. Patterns recognition could be used to distinguish cultivated G. uralensis in Gansu and Xinjiang areas from cultivated and wild plants in Inner Mongolia. Then a total of thirty-one common constituents were identified by LC-TOF/MS analysis coupled with standard compounds information. The contents of four flavonoid glycosides and five saponins were determinated by HPLC and compared using One-way ANOVA. The results showed that there was no significant difference in the contents of 5 triterpenoid saponins among the three regions,but the contents of 4 flavonoid saponins showed the trend of Inner Mongolia >Gansu≈Xinjiang( P<0. 05). In the same Inner Mongolia region,the contents of 4 flavonoid glycosides and 5 triterpenoid saponins in wild plant was significantly higher than that in cultivated plants( P<0. 01). In addition,the contents of liquiritin,isoliquiritin,licorice-saponin A_3,22ß-acetoxyl-glycyrrhizic acid and uralsaponin B in Gansu and Xinjiang were obviously lower than those in Inner Mongolia,but the contents of glycyrrhizic acid,the main component of G. uralensis,were not different in the three geographical regions. In Inner Mongolia,the contents of liquiritin,isoliquiritin,licorice-saponin A_3,licorice-saponin G_2 and glycyrrhizic acid in wild plants were significantly higher than those in cultivated plants. In conclusion,qualitative/quantitative analysis of multi-index components combined with pattern recognition could effectively evaluate the quality of cultivated and wild licorice in different regions. It was helpful for us to understand the reality of licorice in different regions,and provided scientific basis for the development and comprehensive utilization of licorice resources.

[Analysis on the aetiology of influenza in children in Tianjin].

OBJECTIVE: To analyse the pathogen of child patients with influenza in Tianjin area. METHODS: The influenza virus isolation was performed by MDCK cells and embryonated eggs. The identification of the isolates was carried out with hemagglutination (HA) and hemagglutination inhibition (HI) tests. RESULTS: Two hundred and thirty-eight throat swab specimens from children with influenza-like illness were collected in Tianjin area from Oct. 2001 to Mar. 2002 and 64 strains (26.9%) of influenza virus were isolated. Data showed that there were 42 strains (65.6%) of A (H3N2) subtype, 13 strains (20.3%) of A (H1N1) subtype and 9 strains (14.1%) of B type in these positive isolates. All the isolated viruses grew very well in MDCK cells and hemagglutinated with human "O" red blood cells, and most (62/64 strains) of them were able to multiply in embryonated chick eggs. However, there were only 3 isolates with HA positive in inoculating embryonated eggs with the specimens. Meanwhile, it was revealed that out of 55 strains of A type viruses, 53 strains (96.4%) were from O to D phase, 2 strain of A (H3N2) were D phase characters and all B type isolated viruses being D phase properties. CONCLUSION: There were three endemic types of influenza viruses-A (H3N2), A (H1N1) and B type in Tianjin area, with A, the main type.