A UPLC-QTOF/MS-based hepatic tissue metabolomics approach deciphers the mechanism of Huachansu tablets-based intervention against hepatocellular carcinoma.

Huachansu (HCS) tablets, classified as well-known traditional Chinese medicine (TCM) preparation, have been proved to be effective in the treatment of hepatocellular carcinoma (HCC) in clinical studies. However, the underlying mechanism of HCS tablets against HCC has not been comprehensively elucidated. In this study, a rat model of HCC was established with diethylnitrosamine (DEN) inducer. The efficacy of HCS tablets against HCC was assessed through liver histopathological examination and evaluation of biochemical indicators. A metabolomics method based on UPLC-Q-TOF/MS combined with multivariate data analysis was established to identify differential metabolites related to the inhibition effect of HCS tablets on HCC, and then the relevant metabolic pathway analysis was performed to investigate the anti-HCC mechanisms of HCS tablets. The results showed that compared to the control group, the HCC model group showed a significant increase in the values of HCC-related biochemical indicators and the number of tumor nodules, indicating the successful establishment of the HCC rat model. Upon treatment with HCS tablets, the values of HCC-related biochemical indicators decreased, liver fibrosis and nuclear deformation were also significantly alleviated. A total of 15 differential metabolites associated with the anti-tumor effect of HCS tablets on HCC were screened and annotated through hepatic tissue metabolomics studies. Analysis of metabolic pathways revealed that the therapeutic effects of HCS tablets on HCC mainly involved the pentose and glucuronate interconversions and arachidonic acid metabolism. Further western blotting corroborated that the alteration in arachidonic acid (AA) level after the intervention of HCS tablets was related to the inhibition of cPLA2α expression in rat liver tissues. In conclusion, HCS tablets exhibit a certain anti-tumor effect on HCC, and the metabolomics method based on UPLC-Q-TOF/MS combined with further verification at the biochemical level is a promising way to reveal its underlying mechanism.

An integrated analytical strategy to decipher the metabolic profile of alkaloids in Compound Kushen injection based on UHPLC-ESI-QTOF/MSE.

Compound Kushen injection (CKI) is a kind of sterilised water-soluble traditional Chinese medicine preparation that has been used for the clinical treatment of a variety of cancers (hepatocellular carcinoma, lung cancer, etc.) for 19 years. However, to date, the metabolism-related study on CKI in vivo has not been conducted.An integrated analytical strategy was established to investigate the metabolic profile of alkaloids of CKI in rat plasma, urine, and faeces based on ultra-high performance liquid chromatography-electrospray quadrupole time-of-flight mass spectrometry in MSE mode (UHPLC-ESI-QTOF/MSE).Nineteen prototype alkaloids (including 12 matrine-type alkaloids, 2 cytisine-type alkaloids, 3 lupinine-type alkaloids, and 2 aloperine-type alkaloids) of CKI were identified in vivo. Furthermore, 71 metabolites of alkaloids (including 11 of lupanine-related metabolites, 14 of sophoridine-related metabolites, 14 of lamprolobine-related metabolites, and 32 of baptifoline-related metabolites) were tentatively characterised. Metabolic pathways involved in the metabolism of phase I (include oxidation, reduction, hydrolysis, and desaturation), phase II (mainly include glucuronidation, acetylcysteine or cysteine conjugation, methylation, acetylation, and sulphation), and associated combination reactions.The integrated analytical strategy was successfully used to characterise the prototype alkaloids and their metabolites of CKI in vivo, and the results laying a foundation for further study its pharmacodynamic substances.

An ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry method based on a four-step analysis strategy to investigate metabolites of Qi-Yu-San-Long decoction in rat plasma.

Metabolism is undoubtedly significantly correlated with the efficacy and safety of traditional Chinese medicine. In clinic, Qi-Yu-San-Long decoction (QYSLD) has achieved good results in the treatment of non-small-cell lung cancer (NSCLC). Nevertheless, a detailed understanding of the compounds (prototypes and metabolites) of QYSLD and its dynamic metabolic profile in plasma has not been revealed. METHODS: In this study, a rapid and sensitive method based on ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF/MSE ), combined with a four-step analysis strategy, was established to investigate QYSLD metabolic profile in rat plasma. RESULTS: In all, 101 xenobiotics (41 prototypes and 60 QYSLD-related metabolites) were identified in rat plasma. The research uncovered metabolic profiles of alkaloids, saponins, flavonoids, iridoids, anthraquinones, and phenylpropanoids of QYSLD in rat plasma. The dynamic changes in these xenobiotics were also observed at different time intervals. At 0.5 h after oral administration, only 15 prototypes and 11 metabolites were detected. Within 24 h, 4 prototypes and 20 metabolites can still be detected. Four prototypes and 10 metabolites had the phenomenon of emergence-disappearance-reappearance in vivo. CONCLUSION: In rat plasma, 101 xenobiotics of QYSLD were identified and their dynamic metabolic profiles were systematically delineated, which laid a material basis for further research of the pharmacodynamic substances of QYSLD inhibiting NSCLC.

Discovering the potential active ingredients of Qi-Yu-San-Long decoction for anti-oxidation, inhibition of non-small cell lung cancer based on the spectrum-effect relationship combined with chemometric methods.

Qi-Yu-San-Long decoction (QYSLD), a traditional Chinese medicine (TCM) prescription, consisting of ten types of herbal medicine which has significant clinical efficacy in the treatment of non-small cell lung cancer (NSCLC). However, the bioactive ingredients of QYSLD remain unclear, due to their "multi-ingredients" and "multi-targets" features. This study aimed to construct a spectrum-effect correlation analysis model and screen the potential active components of QYSLD. A fingerprint method based on ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) was developed and validated to obtain seventy common peaks of ten batches of QYSLD. The results of methodological evaluation, including precision, repeatability and stability, were less than 8.19%. In terms of linearity, eleven common components did not reach the linear standard (R2 < 0.99), they were removed before spectrum-effect relationship analysis. After treated with ten batches of QYSLD, the results of DPPH and FRAP assays ranged from 1.59 to 5.50 mg mL-1 and 143.83-873.83 µmol L-1, respectively. Meanwhile, the cell viabilities of A549 cells treated with QYSLD samples ranged from 21.73% to 85.71%. The relative healing rates ranged from 21.50% to 44.46%. The number of migrated and invaded cells ranged from 12.00 to 68.67 and 7.67 to 27.00, respectively. Then, the potential active components of QYSLD were screened through spectrum-effect relationship constructed by grey correlation analysis (GRA), partial least squares regression (PLSR) and backpropagation neural network (BP-ANN). The results were as follow: 1) eight ingredients of QYSLD were relevant to DPPH free radical scavenging ability; 2) nine ingredients were relevant to FRAP; 3) six ingredients were relevant to inhibit the proliferation ability of A549 cells; 4) twenty-two ingredients were relevant to inhibit the horizontal migration ability; 5) five ingredients were relevant to inhibit the vertical migration ability; 6) twelve ingredients were relevant to inhibit the invasion ability. Confirmatory experiments showed that compared with the unscreened ingredients, the potential active ingredients screened by the spectrum-effect relationship had better antioxidant and anti-NSCLC effects. In general, this study found the potential active ingredients in QYSLD. Meanwhile, the established method provided a valuable reference model for the potential active ingredients of TCM.

The dynamic metabolic profile of Qi-Yu-San-Long decoction in rat urine using UPLC-QTOF-MSE coupled with a post-targeted screening strategy.

Qi-Yu-San-Long decoction (QYSLD) is a traditional Chinese medicine that has been clinically used in the treatment of non-small-cell lung cancer (NSCLC) for more than 20 years. However, to date, metabolic-related studies on QYSLD have not been performed. In this study, a post-targeted screening strategy based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight full information tandem mass spectrometry (UPLC-QTOF-MSE) was developed to identify QYSLD-related xenobiotics in rat urine. The chemical compound database of QYSLD constituents was established from previous research, and metabolites related to these compounds were predicted in combination with their possible metabolic pathways. The metabolites were identified by extracted ion chromatograms using predicted m/z values as well as retention time, excimer ions, and fragmentation behavior. Overall, 85 QYSLD-related xenobiotics (20 prototype compounds and 65 metabolites) were characterized from rat urine. The main metabolic reactions and elimination features of QYSLD included oxidation, reduction, decarboxylation, hydrolysis, demethylation, glucuronidation, sulfation, methylation, deglycosylation, acetylation, and associated combination reactions. Of the identified molecules, 14 prototype compounds and 58 metabolites were slowly eliminated, thus accumulating in vivo over an extended period, while five prototypes and two metabolites were present in vivo for a short duration. Furthermore, one prototype and five metabolites underwent the process of "appearing-disappearing-reappearing" in vivo. Overall, the metabolic profile and characteristics of QYSLD in rat urine were determined, which is useful in elucidating the active components of the decoction in vivo, thus providing the basis for studying its mechanism of action.

An integrated strategy to delineate the chemical and dynamic metabolic profile of Huachansu tablets in rat plasma based on UPLC-ESI-QTOF/MSE.

Metabolic research is a key method to understand the fate of traditional Chinese medicine (TCM) prescription in vivo and help to explain its pharmacological effects. Huachansu (HCS) tablets are prepared from the dried skin of Bufo gargarizans (Cantor, 1842), which have the effects of detumescence and analgesia, and used in the treatment of middle and advanced tumors. However, an in-depth understanding of the chemical components of HCS tablet and its metabolism in vivo is lacking. In this study, an integrated analytical strategy based on UPLC-ESI-QTOF/MSE was developed to efficiently identify the chemical components, the absorbed prototype compounds and metabolites of HCS tablets given by gavage in rats, and track their dynamic changes. As a result, 77 chemical components of HCS tablets were characterized, including 46 bufadienolides, 11 alkaloids, 10 amino acids and organic acids, 9 nucleosides, and 1 other component. Further more, 16 prototype compounds and 47 metabolites (38 bufadienolides-related and 9 alkaloids-related) were identified in rat plasma. Through the dynamic detection of prototype compounds and metabolites of HCS tablets in vivo, the phenomenon that 3 prototype compounds and 16 metabolites reappeared after a period of disappearance from the plasma was found. Additionally, 8 prototype compounds with large intensity in HCS tablet samples were not detected in the plasma samples of rats given HCS tablets but had metabolites within 0-24 h after administration, indicating that these components were rapidly absorbed and metabolized into metabolites in vivo. It was also observed that 9 metabolites can be driven from various prototype compounds and reach high concentrations in a short time. This study comprehensively identified the xenobiotics and metabolites of HCS tablets in rat plasma sample, systematically discussed their dynamic metabolic process in vivo, which laid a foundation for studying the pharmacodynamic substances of HCS tablets.

Sustainable management of cadmium-contaminated soils as affected by exogenous application of nutrients: A review.

Cadmium (Cd) pollution in arable land is of great concern as it impairs plant growth and further threats human health via food-chain. Exogenous supplementation of nutrients is an environmentally-friendly, cost-effective, convenient and feasible strategy for regulating Cd uptake, transport and accumulation in plants. To sustain Cd-contaminated soils management, on the one hand, a low level of the Cd-contaminated soil is expected to cultivate crops with decreased Cd accumulation as affected by exogenous nutrients application, on another hand, a high level of the Cd-contaminated soil is suggested to cultivate phytoextraction plants with increased Cd accumulation as affected by exogenous nutrients application. Nevertheless, effects of nutrients on Cd accumulation in plants are still ambiguous. Thus, data of Cd accumulation in shoots of plants as affected by exogenous application of nutrients were collected from previously published articles between 2005 and 2021 in the present study. According to the data, exogenous supply of calcium (Ca), magnesium (Mg), iron (Fe), manganese (Mn) and silicon (Si) to a larger extent decrease Cd amounts in shoots of plants. By contrast, exogenous nitrogen (N), and deficient Ca, Mg and Fe supply have a great possibility to increase Cd amounts in shoots of plants. Although exogenous application of phosphorus (P), sulfur (S), potassium (K), zinc (Zn) and selenium (Se) have a great opportunity to increase biomass, they show different effects on Cd concentrations. As a result, the odds are even for increasing and decreasing Cd amounts in shoots of plants. Taken together, exogenous application of Ca, Mg, Fe, Mn and Si might decrease Cd accumulation in plants that are recommended for crops production. Exogenous N and deficient Ca, Mg and Fe supply might increase Cd accumulation in plants that are recommended for phytoextraction plants. Exogenous application of P, S, K, Zn and Se have half a chance to increase or decrease Cd accumulation in plants. Therefore, dosages, forms and species should be taken into account when exogenous P, S, K, Zn and Se are added.

Nutrient inputs from the leaf decay of Cynodon dactylon (L.) Pers in the water level fluctuation zone of a Three Gorges tributary.

Cynodon dactylon (L.) Pers (C. dactylon) is one of the dominant plants in the water level fluctuation (WLF) zone of the Three Gorges Reservoir (TGR) tributaries. However, the leaves of C. dactylon can decay to increase the inputs of nutrients under flood inundation, increasing the risk of eutrophication in the TGR tributaries. Nutrient inputs from the leaf decay of C. dactylon in three interfaces, namely, water-sediment (WS), water-C. dactylon (WC) and water-sediment-C. dactylon (W-S-C), were estimated in a 180 d inundation experiment. The results showed that the kinetic processes of total dissolved nitrogen (TDN) and total dissolved phosphorus (TDP) input accorded with the power function equation: y = axb for the WS, WC and W-S-C interfaces (R2s > 0.72, p < 0.001). The cumulative TDN input from leaf decay of C. dactylon in the WC interface was 506.44 mg N kg-1 of biomass, which was significantly higher than that in the W-S-C interface with 422.24 mg N kg-1 of biomass (p < 0.05). However, no significant differences in TDP input were found between the WC and W-S-C interfaces (p > 0.05). The total amounts of TDN and TDP inputs at the 165-175 m altitude were 21,688.81 and 13,121.68 kg year-1, respectively, which were approximately 3.17 times those from the 145-155 m altitude of the WLF zone. The amounts of TDN and TDP inputs from the leaves of C. dactylon for the whole WLF zone were 49,261.65 and 29,803.17 kg year-1, respectively, which were 0.1 and 2.7 times the annual permissible discharge amount of pollutants calculated from a municipal wastewater treatment plant with the peak flow of 60,000 m3/d according to Class I (A) of the Wastewater Discharge Standard (GB18918-2002) in China. Thus, the aboveground part of this perennial herb should be harvested in a timely manner before reflooding, especially at the higher altitudes of the WLF zone to decrease eutrophication risk.

Icaritin suppresses multiple myeloma, by inhibiting IL-6/JAK2/STAT3.

Icaritin is an active prenylflavonoid derived from Epimedium genus, a traditional Chinese medicine. Icaritin has a wide range of pharmacological and biological activities, including cardiovascular function improvement, hormone regulation and antitumor activity. Here, we investigated the effect of icaritin on multiple myeloma (MM) in vitro and in vivo. Icaritin inhibited cell growth of MM cell line and primary MM cells. In contrast, icaritin had low or no cytotoxic effect on normal hematopoiesis. We also demonstrated that in MM xenograft mouse models, icaritin suppressed tumor growth and decreased serum IL-6 and IgE levels, but did not show adverse reactions such as body weight loss. The anti-MM activity of icaritin was mainly mediated by inhibiting IL-6/JAK2/STAT3 signaling. We suggest that icaritin can be further tested in clinical trials in MM.

Discovery andw biological evaluation of novel 6,7-disubstituted-4-(2-fluorophenoxy)quinoline derivatives possessing 1,2,3-triazole-4-carboxamide moiety as c-Met kinase inhibitors.

A series of 6,7-disubstituted-4-(2-fluorophenoxy)quinoline derivatives possessing 1,2,3-triazole-4-carboxamide moiety were designed, synthesized and evaluated for their in vitro biological activities against c-Met kinase and five typical cancer cell lines (A549, H460, HT-29, MKN-45 and U87MG). Most compounds showed moderate to excellent antiproliferative activity. In this study, a promising compound 34, with a c-Met IC50 value of 1.04nM, was identified as a multitargeted receptor tyrosine kinase inhibitor. The SAR analyses indicated that compounds with halogen group, especially fluoro group, at 4-position on the phenyl ring (moiety B) have potent antitumor activity, and methylation on the 5-atom linker played an important role in the c-Met enzymatic activity.

Notolutesins A-J, dolabrane-type diterpenoids from the Chinese liverwort Notoscyphus lutescens.

Ten new dolabrane-type diterpenoids, notolutesins A-J (1-10), were isolated from the Chinese liverwort Notoscyphus lutescens, along with four known compounds. The structures of the new compounds were established on the basis of extensive spectroscopic data, and that of 1 was confirmed by single-crystal X-ray crystallography. The absolute configuration of 1 was determined by comparing its experimental and calculated electronic circular dichroism spectra. All of the isolates were evaluated for their cytotoxicity against a small panel of human cancer cell lines, and compound 1 exhibited an IC50 value of 6.2 µM against the PC3 human prostate cancer cell line.

Phytotoxic cis-clerodane diterpenoids from the Chinese liverwort Scapania stephanii.

Five cis-clerodane diterpenoids, stephanialides A-E, along with seven known cis-clerodanes, scaparvins A-C, parvitexins B and C, 3-chloro-4-hydroxy-parvitexin A, and scapanialide B, were isolated from the Chinese liverwort Scapania stephanii. Their structures were established unequivocally on the basis of spectroscopic data. The absolute configuration of stephanialide A was determined by analysis of CD data using the octant rule. Phytotoxic activity evaluation showed that this type of diterpenoids can significantly inhibit root elongation of the seeds of Arabidopsis thaliana, Lepidium sativum and Brassica pekinensis.

Marsupellins A-F, ent-longipinane-type sesquiterpenoids from the Chinese liverwort Marsupella alpine with acetylcholinesterase inhibitory activity.

Acetylcholinesterase (AChE) inhibitory activity-guided fractionation of the Chinese liverwort Marsupella alpine afforded six new [marsupellins A-F (1-6)] and three known (7-9) ent-longipinane-type sesquiterpenoids. The structures were determined from MS and NMR spectroscopic data, single-crystal X-ray diffraction, and electronic circular dichroism calculations. Compounds 1-9 exhibited moderate to weak AChE inhibitory activity.

Discovery of novel 6,7-disubstituted-4-phenoxyquinoline derivatives bearing 5-(aminomethylene)pyrimidine-2,4,6-trione moiety as c-Met kinase inhibitors.

A series of novel quinoline derivatives bearing 5-(aminomethylene)pyrimidine-2,4,6-trione moiety were designed, synthesized, and evaluated for their c-Met kinase inhibitory activities and antiproliferative activities against 5 cancer cell lines (HT-29, H460, MKN-45, A549, and U87MG) in vitro. Most compounds showed moderate to excellent potency, with the most promising analogue 45 (c-Met half-maximal inhibitory concentration [IC50]=1.15 nM) showing high selectivity versus 5 other tyrosine kinases, VEGFR-2, Flt-3, PDGFR-ß, c-Kit, and EGFR. Structure-activity relationship studies indicated that electron-donating groups on the phenyl ring at the 3-position of pyrimidine-2,4,6-trione were required to increase the electron density on the 5-(aminomethylene)pyrimidine-2,4,6-trione moiety.

Secondary metabolites from the Chinese liverwort Cephaloziella kiaeri.

Sixteen new clerodane diterpenoids, cephaloziellins A-P (1-16), and two known analogues (17 and 18) were isolated from an EtOH extract of the Chinese liverwort Cephaloziella kiaeri. The structures of the new compounds were elucidated from extensive spectroscopic data (IR, UV, HRESIMS, 1D NMR, and 2D NMR), and the structures of 5, 9, and 15 were confirmed by single-crystal X-ray diffraction analyses. The absolute configurations of all new compounds were established by comparing experimental and calculated electronic circular dichroism spectra.

Highly oxygenated ent-pimarane-type diterpenoids from the Chinese liverwort Pedinophyllum interruptum and their allelopathic activities.

Ten highly oxygenated ent-pimarane-type diterpenoids, pedinophyllols A-J (1-10), were isolated from the Chinese liverwort Pedinophyllum interruptum. Their structures were determined by comprehensive analysis of spectroscopic data together with single-crystal X-ray diffraction analysis. The absolute configurations were elucidated by comparison of experimental and theoretically calculated electronic circular dichroism spectra. Allelopathic testing showed that several new diterpenoids inhibited germination of Arabidopsis thaliana seeds.

Terpenoids from the Chinese liverwort Plagiochila pulcherrima and their cytotoxic effects.

Three new pimarane-type diterpenoids, 7ß,11α-dihydroxypimara-8(14),15-diene (1), 1ß,11α-dihydroxypimara-8(14),15-diene (2), and 11α-hydroxypimara-8(14),15-diene (3), five 2,3-secoaromadendrane-type sesquiterpenoids, including a new one, ethyoxyplagiochiline A2 (4), and three known fusicoccane-type diterpenoids were isolated from the Chinese liverwort Plagiochila pulcherrima. Their structures were established on the basis of extensive spectroscopic analysis. Compounds 6 and 7 exhibited moderate inhibitory activity on the proliferation of human cancer cell lines Hela, A172, and H460. Moreover, the cytotoxicity of compound 6 to Hela cells was related to apoptotis as confirmed by DAPI nuclear staining and flow cytometry.