This study aimed to explore gene expression profiles that drive malignancy from low- to high-grade head and neck carcinomas (HNC), as well as to analyze their correlations with survival. Gene expressions and clinical data of HNC were downloaded from the Gene Expression Omnibus (GEO) repository. The significantly differential genes (SDGs) between low- and high-grade HNC were screened. Cox regressions were performed to identify prognostic SDGs of progression-free survival (PFS) and disease-specific survival (DSS). The genes were experimentally validated by RT-PCR in clinical tissue specimens. Thirty-five SDGs were identified in 47 low-grade and 30 high-grade HNC samples. Cox regression analyses showed that CXCL14, SLC44A1, and UBD were significantly associated with DSS, and PPP2R2C and SLC44A1 were associated with PFS. Patients were grouped into high-risk or low-risk groups for prognosis based on gene signatures. High-risk patients had significantly shorter DSS and PFS than low-risk patients (P=0.033 and P=0.010, respectively). Multivariate Cox regression showed HPV (P=0.033), lymph node status (P=0.032), and residual status (P<0.044) were independent risk factors for PFS. ROC curves showed the risk score had better efficacy to predict survival both for DSS and PFS (AUC=0.858 and AUC=0.901, respectively). The results showed CXCL14 and SLC44A1 were significantly overexpressed in the low-grade HNC tissues and the UBD were overexpressed in the high-grade HNC tissues. Our results suggested that SDGs had different expression profiles between the low-grade and high-grade HNC, and these genes may serve as prognostic biomarkers to predict survival.

Abstract This study investigated the influence of different processing methods on the oral toxicity of Sophora alopecuroides L., Fabaceae, seeds in mice and on the contents of five known toxic-effective quinolizidine alkaloids from the ethanol extracts quantified by ultra-performance liquid chromatography coupled to tandem quadrupole mass spectrometry. It provides an evidence to elucidate the possible reasons why vinegar-processing and parching methods significantly decrease the acute oral toxicity induced by S. alopecuroides and why wine-processing method increases it instead (demonstrated by measurement of LD50 and histopathological analysis). The analytical performance for the determination of the five analytes was evaluated by linearity, stability, repeatability, precision and accuracy, and recovery test. The lowest limit of quantification was determined to be 5 ng/ml for each substance and the precision and accuracy at lowest limit of quantification were below 20%. Cytisine, the most toxic alkaloid among the five alkaloids, declined 11.26, 3.98, and 2.73 folds after being vinegar-processed and fried in a ceramic or iron pan, respectively and had a very close correlation with the toxicity of S. alopecuroides seeds (r = 0.8589). Other matrine-type alkaloids with lower toxicity including matrine, sophcarpine, and sophoridine decreased after being wine-processed and fried in a ceramic pan, but increased 4.44, 7.20, and 7.23 folds when being processed by vinegar. Oxymatrine declined in all groups. It, therefore, reveals that vinegar-processing method reduces the oral toxicity of S. alopecuroides mainly due to a sharp decrease of cytisine, thus improves its clinical safety.

Abstract Kudouzi (Sophora alopecuroides L., Fabaceae) is an effective folk medicine, but it always causes a hepatic and renal toxicity in clinical therapy. The toxic mechanism remains unclear. This paper detected the urinary and plasma metabolites alteration by 1H NMR-based metabonomics study in Kudouzi-induced rats to evaluate the toxic mechanism for clinical security. The male Sprague-Dawley rats were orally dosed with 0.5 and 1 g Kudouzi/kg weight once per day for consecutive 14 days. Urine samples were collected at day −1 (before treatment), and days 7, 14, and 21 for NMR analysis, respectively. Plasma samples were harvested at day 14 for NMR and biochemical analysis. The metabonomic profiling of Kudouzi-treated rats differed from that of the vehicle. This was confirmed by the biochemistry analysis. The accumulated subacute toxicity of Kudouzi was visible with dosing time, and persisted at day 21 even after the disposal was ended. The observable biochemical pathways alterations included inhibited TCA cycle, activated anaerobic glycolysis, perturbed amino acids metabolism, and disordered gut microbiota. The results evidenced the toxicity mechanism of Kudouzi from a systematic and holistic view.

Abstract: Chuju, the flower of Chrysanthemum morifolim cv. Chuju, is a traditional medicine-food material used in China. Bioassay-guided chemical investigation on Chuju led the isolation of 16 flavonoids (1-16) characterized as quercetin (1), isorhamnetin 3-O-b-D-glucoside (2), eriodictyol (3), pyracanthoside (4), apigenin (5), apigetrin (6), acacetin (7), acacipetalin (8), luteolin (9), diosmetin (10), spinacetin (11), axillarin (12), bonanzin (13), cirsiliol (14), chrysosplenol D (15) and artemetin (16). Among them, compounds (11-16) were first reported from the Chrsanthemum species. Chuju flavonoids displayed strong antioxidant in DPPH (1, 1-diphenyl-2- picrylhydrazyl) radical scavenging assay, and high anti-inflammatory activity by inhibiting NO production in LPS-induced RAW 264.7 cells. These findings suggests that Chuju is an ideal source of natural flavonoids with significant antioxidant and anti-inflammatory effects, and is worthy of further development as health-promoting food or supplement.

Resumen: La flor de Chrysanthemum morifolim cv. Chuju, conocida popularmente como Chuju, se utiliza tradicionalmente como alimento y medicina en China. Un estudio químico biodirigido de un extracto de las flores condujo al aislamiento de 16 flavonoides, los cuales se caracterizaron como quercetina (1), isoramnetina 3-O-b-D-glucosido (2), eriodictiol (3), piracantosido (4), apigenina (5), apigetrina (6), acacetina (7), acacipetalina (8), luteolina (9), diosmetina (10), espinacetina (11), axilarina (12), bonanzina (13), cirsiliol (14), crisosplenol D (15) and artemetina (16). Los productos 11-16 se describen por primera vez en la especie. Los flavonoides de Chuju demostraron una notable actividad antioxidante en el ensayo de captación del radical 2,2- difenil-1-picrilhidrazilo (DPPH); asimismo presentaron un fuerte efecto antiinflamatorio in vitro, valorado a través de la inhibición de la producción de NO en macrófagos RAW 264.7 estimulados con LPS. Estos resultados indican que Chuju es una fuente importante de flavonoides antiinflamatorios y antioxidantes con un potencial importante para su desarrollo como agente nutracéutico.

ABSTRACT Our previous work revealed that chrysosplenetin in combination with artemisinin inhibited in vivo P-glycoprotein (P-gp, one of classic multi-drug resistance proteins) mediated digoxin transportation activity by reversing the upregulated P-gp/Mdr1 mRNA expression levels by artemisinin. Therefore, chrysosplenetin might be a potential artemisinin-resistance reversal agent as a P-gp inhibitor. But it still remains unknown if chrysosplenetin has an impact on another pivotal multi-drug resistance protein, breast cancer resistance protein (Bcrp), which is co-expressed with P-gp in apical membrane of intestinal epithelial cell and overlaps some of the substrates and inhibitors. This study, therefore, further addressed the impact of chrysosplenetin, per se or in combination with artemisin, on Bcrp/ABCG2 mRNA expression levels in mice small intestine determined by western blot and real time-quantitative polymerase chain reaction (RT-qPCR) assay. The drugs were intragastrically administrated once per day for 7 days. Novobiocin, a known Bcrp inhibitor, was observed to have no impact on Bcrp/ABCG2 levels with or without artemisinin versus vehicle. Interestingly, artemisinin alone attenuated Bcrp level while chrysosplenetin alone increased it (p < 0.05). Relative mRNA level was significantly decreased when co-used with artemisinin and chrysosplenetin in ratio of 1:2 (p < 0.05). The discrepant results for chrysosplenetin on Bcrp/ABCG2 mRNA expressions might be closely related to the transcriptional or posttranscriptional regulation.

Abstract: A new sesquilignan effususin E (1) and a new long chain fatty enamide effususin F (2), together with eight known compounds were isolated from the medullae of Juncus effusus. The structures of 1 and 2 were elucidated by detailed spectroscopic analysis including 1H and 13C NMR, COSY, HSQC, HMBC, and HRMS, while the structures of the known compounds were deduced from comparison of their spectral data with those in the literature. The new compounds 1 and 2 were evaluated against five human cancer cell lines (SHSY-5Y, SMMC-7721, HepG-2, Hela, and MCF-7) by CCK-8 assay, but only 1 exhibited weak cytotoxic activity against SMMC-7721 with IC50 value of 57.5 μm. Inhibitory effects of 1 and 2 on nitric oxide (NO) production in lipopolysaccharide-induced RAW264.7 macrophages were also evaluated, and they were found to exhibit good inhibitory activities with IC50 values of 8.59 and 13.73 μm, respectively.

Resumen: Se elucidaron las estructuras de 1 y 2 mediante análisis espectrocópicos de RMN de 1H y 13C, COSY, HSQC, HMBC, y HRMS, mientras que las estructuras de los compuestos conocidos se dedujeron por comparación de sus datos espectrales con los de la literatura. Los nuevos compuestos 1 y 2 fueron evaluados contra cinco líneas celulares cancerígenas humanas (SHSY-5Y, SMMC-7721, HepG-2, Hela, and MCF-7) mediante el ensayo CCK-8. Solamente el compuesto 1 mostró una débil actividad citotóxica contra la línea SMMC-7721 con un valor de IC50 de 57.5 μM. También se evaluó el efecto inhibitorio de los compuestos 1 y 2 en la producción de óxido nítrico (NO) en macrófagos RAW264.7 inducidos por lipopolisacáridos. Ambos compuestos mostraron buena actividad inhibitoria con valores de IC50 de 8.59 y 13.73 μM, respectivamente.

Chemical investigation on the root of R. laevigata led the isolation and identification of two new flavonoids, (+)-catechin-8-acetic acid (1) and guibourtacacidine 4-methyl ether (2), one known flavonoid, guibourtacacidine (3), along with seven known triterpenoids, euscaphic acid (4), kajiichigoside F1 (5), nigaichigoside F2 (6), rubuside J (7), tomentic acid (8), rosamutin (9) and betulinic acid (10). Their structures were established on the basis of spectroscopic evidence and comparisons with literature data. Compounds 1, 2 and 3 showed considerable radical scavenging activity in the 1, 1-diphenyl-2-picrylhydrazyl (DPPH) assay.

La investigación química de las raíces de R. laevigata condujo al aislamiento de dos nuevos flavonoides, 8-(+)-catequin-ácido acético (1) y éter 4-metílico de guibourtacacidina (2); un flavonoide conocido, guibourtacacidina (3), y siete triterpenos conocidos, ácido euscáfico (4), kajichi-gósido F1 (5), nigaichigósido F2 (6), rubúsido J (7), ácido toméntico (8), rosamutina (9) y ácido betulínico (10). Las estructuras fueron establecidas de acuerdo a las evidencias espectroscópicas y comparaciones con los datos informados en la literatura. Los compuestos 1, 2 y 3 mostraron actividad atrapadora de radicales en el ensayo del 1,1-difenil-2-picrilhidracilo (DFPH).