Combination of curcumin with N-n-butyl haloperidol iodide inhibits hepatocellular carcinoma malignant proliferation by downregulating enhancer of zeste homolog 2 (EZH2) - lncRNA H19 to silence Wnt/ß-catenin signaling.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common cause of cancer-related death worldwide. Curcumin (C) has been extensively investigated in different types of malignancies, including hepatocellular carcinoma, but its physicochemical properties have significantly influenced its clinical use. Several approaches are being explored to enhance curcumin's therapeutic response, including its combination with various drugs. PURPOSE: This study aimed to evaluate the anti-tumor effect of curcumin (C) in combination with F2 (N-n-butyl haloperidol iodide) on hepatocellular carcinoma and its potential underlying mechanism in vitro and in vivo. METHODS: Cell proliferation was evaluated by CCK-8 and colony formation assays, and apoptosis was measured by flow cytometry. The migratory and invasive abilities of Hep3B and SMMC-7721 cells were measured by wound-healing and matrigel transwell assays. In order to investigate the molecular pathways, various experiments such as western blotting, qPCR, RNA-seq, immunostaining and transfection were performed. To evaluate the anti-HCC effects in vivo, a xenograft tumor model was used. RESULTS: Our findings showed that the combination of curcumin (C) & F2 (F2C) strongly inhibited malignant proliferation and migration in SMMC-7721 and Hep3B cells. The F2C treatment downregulates enhancer of zeste homolog 2 (EZH2) transcription and protein expression, which is key epigenetic regulator responsible for HCC development. Moreover, the inhibition of EZH2 by F2C led to Wnt/ß-catenin signaling inhibition by decreasing tri-methylation of histone H3 at lysine 27 (H3K27me3) and long non-coding RNA H19 expression. The inhibition of F2C was associated with the suppression of tumorigenicity in xenograft HCC models. CONCLUSION: These findings suggested that, F2C inhibited HCC formation, migration and its modulatory mechanism seemed to be associated with downregulation of EZH2, silencing Wnt/ß-catenin signaling by interacting with H19, suggesting that F2C may be a promising drug in the clinical treatment of HCC.

Arsenic Bioaccessibility of Realgar Influenced by the Other Traditional Chinese Medicines in Niuhuang Jiedu Tablet and the Roles of Gut Microbiota.

Niuhuang Jiedu tablet (NJT), a realgar (As2S2) containing Traditional Chinese Medicine (TCM), is a well-known formula. The safety of NJT is of growing concern since arsenic (As) is considered as one of the most toxic elements. NJT was demonstrated to be safer than realgar by our previous experiments and some other studies. The toxicity of realgar has been shown to be related to the amount of soluble or bioaccessible arsenic. In this study, the influences of the other TCMs in NJT on the bioaccessibility of arsenic from realgar, and the roles of gut microbiota during this process were investigated in vitro. Results showed that Dahuang (Rhei Radix et Rhizoma), Huangqin (Scutellariae Radix), Jiegeng (Platycodonis Radix), and Gancao (Glycyrrhizae Radix et Rhizoma) could significantly reduce the bioaccessibility of arsenic from realgar in artificial gastrointestinal fluids. Gut microbiota played an important role in decreasing the bioaccessibility of realgar because it was demonstrated to be able to absorb the soluble arsenic from realgar in the incubation medium. Dahuang, Huangqin, and Jiegeng could modulate the gut microbiota to enhance its arsenic absorption activity.

Screening and quantification of 18 glucocorticoid adulterants from herbal pharmaceuticals and health foods by HPLC and confirmed by LC-Q-TOF-MS/MS.

A procedure for the screening and quantification of 18 glucocorticoids, i.e. hydrocortisone sodium succinate (HSS), prednisolone (PDL), prednisone (PDS), hydrocortisone (HCS), methylprednisolone (MPS), betamethasone (BTM), dexamethasone (DXM), triamcinolone acetonide (TA), prednisolone acetate (PLA), hydrocortisone acetate (HA), fludrocortisone acetate (FA), prednisone acetate (PA), cortisone acetate (CA), dexamethasone acetate (DA), hydrocortisone butyrate (HB), triamcinolone acetonide acetate (TAA), fluocinonide (FN) and halcinonide (HC) from herbal pharmaceuticals and health foods was established and fully validated. The samples were extracted by methanol and separated by HPLC. The retention times and ultraviolet spectra were used for the preliminary screening, and the suspected adulterants were then confirmed by liquid chromatography-quadrupole-time of flight mass spectrometry (LC-Q-TOF-MS/MS) and quantified by HPLC. The developed procedure was successfully applied to 14 herbal samples, and 316.3 µg g-1 of DA and 13.6 µg mL-1 of BTM were found in a tablet sample and a spray sample, respectively. To our best knowledge, this is the first report of the simultaneous screening and quantification of these 18 glucocorticoids from any matrix.

Laccase Production Among Medicinal Mushrooms from the Genus Flammulina (Agaricomycetes) Under Different Treatments in Submerged Fermentation.

The laccase activities of 13 strains of medicinal and edible mushrooms from the Flammulina genus (F. velutipes, F. rossica, and F. fennae) were studied. The effects of both fungal isolates and culture media were investigated. The laccase activities indicated significant differences among Flammulina strains (P < 0.001), and the cultural media significantly affected the laccase activities in Flammulina spp. (P < 0.001). The morphological characteristics of the 3 Flammulina species were similar, but differences in phylogenetic analysis and laccase activity existed among different species. Although isolates of each Flammulina species differed, the laccase variables among different Flammulina species were greater than those within the same species. The presence of simple carbon and nitrogen sources increased the maximum laccase enzyme activity, but the occurrence of both laccase activity and maximum laccase enzyme activity was delayed compared with lignocellulosic material. The laccase activities of most Flammulina isolates were increased when exposed to copper ions. Our findings offer insights into laccase productivity in response to different Flammulina species or strains and different submerged fermentation treatments.

[Study on arsenic speciation changes in crude and processed traditional Chinese medicines by HPLC-ICP-MS].

A HPLC-ICP-MS method for simultaneous determination of As(III), As(V), MMA and DMA in traditional Chinese medicines (TCMs) was established, and the contents of As(III), As(V), MMA and DMA in a TCM with high total arsenic content (Cordyceps) and 5 crude and processed TCMs (Radix Astragali, Radix et Rhizoma Rhei, Radix Scutellariae, Radix Polygoni Multiflori and Radix Rehmanniae) were determined and analyzed. The method validation indicated that the correlative coefficients (r) for all speciations were bigger than 0.9984; the limits of quantitation (LOQ) were from 0.8 to 1.0 microg x L(-1); the reproducibility and stability were satisfactory with all RSDs less than 10%; the spiked recoveries ranged from 82.40% to 119.5%. The results of samples analysis showed that the inorganic arsenic (As(III) and As(V)) was the dominating speciation in the tested TCMs; MMA and DMA were not found in all plant resourced TCMs, but MMA was found in Cordyceps; all the tested TCMs indicated a content increasing of inorganic arsenic after processing.