The bis-Schiff base of N,N′-1,10-bis(naringin) triethylenetetraamine (1) was prepared, as a copper(II) ion chelator, compound 1 was used for Alzheimer’s disease therapy in vitro. The ...3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay of compound 1 showed that this Schiff base could promote PC12 cells proliferation, and also, compound 1 could inhibit Cu2+-amyloid-β (Aβ)1–42 mediated cytotoxicity on PC12 cells. The thioflavine T (ThT) assay showed that 1 can effectively attenuate Cu2+-induced Aβ1–42 aggregation. In addition, compound 1 is determined to be potent antioxidants on the basis of in vitro antioxidant assay, it can effectively decease the level of reactive oxygen species (ROS) in Cu2+-Aβ1–42-treated PC12 cells and elevate the superoxide dismutase (SOD) activity in Cu2+-Aβ1–42-treated PC12 cells. The results show that N,N′-1,10-bis(naringin) triethylenetetraamine is a potential agent for therapy of Alzheimer’s disease.
Gambogic acid (GA) is a potential anti-cancer compound that is extracted from the resin of Garciania hanburyi. The present study was designed to evaluate the anti-metastatic effect of GA on melanoma ...cell lines in vitro and to explore the underlying mechanism. The anti-proliferative activity of GA on melanoma cells was assessed by CCK-8 assay. The Wound-healing, transwell, adhesion, and tube formation assays were performed to examine the inhibition of GA on the cell's migration, invasion, adhesion, and angiogenesis capacities, respectively. Enzymatic activity of MMP-2 and MMP-9 were detected by gelatin zymography assay. Protein expressions regulated by GA treatment were tested by Western blot assay. The present results showed that GA significantly inhibited the proliferation of highly metastatic melanoma A375, B16-F10 cells and human umbilical vein endothelial cells (HUVECs) in time- and doses-dependent manners. Furthermore, GA significantly inhibited the migratory, invasive and adhesive properties of A375 and B16-F10 cells, and tube-forming potential of HUVECs at sub-IC50 concentrations, where no significant cytotoxicity was observed. Mechanistically, GA treatment suppressed the EMT and angiogenesis processes and reduced the enzymatic activity of MMP-2 and MMP-9. Moreover, abnormal PI3K/Akt and ERK signaling pathways in A375 and B16-F10 cells and HUVECs were notably suppressed by GA treatment. Collectively, our results suggest that GA exerts anti-metastasis activity in melanoma cells by suppressing the EMT and angiogenesis through the PI3K/Akt and ERK signaling pathways, and might be used as a phytomedicine against metastatic melanoma.
Display omitted
Objective
This study intended to establish a droplet digital PCR (dd‐PCR) for monitoring minimal residual disease (MRD) in patients with BCR/ABL (P210)‐positive chronic myeloid leukemia (CML), ...thereby achieving deep‐level monitoring of tumor load and determining the efficacy for guided clinically individualized treatment.
Methods
Using dd‐PCR and RT‐qPCR, two cell suspensions were obtained from K562 cells and normal peripheral blood mononuclear cells by gradient dilution and were measured at the cellular level. At peripheral blood (PB) level, 61 cases with CML‐chronic phase (CML‐CP) were obtained after tyrosine kinase inhibitor (TKI) treatment and regular follow‐ups. By RT‐qPCR, BCR/ABL (P210) fusion gene was undetectable in PB after three successive analyses, which were performed once every 3 months. At the same time, dd‐PCR was performed simultaneously with the last equal amount of cDNA. Ten CML patients with MR4.5 were followed up by the two methods.
Results
At the cellular level, consistency of results of dd‐PCR and RT‐qPCR reached R2 ≥ 0.99, with conversion equation of Y = 33.148X1.222 (Y: dd‐PCR results; X: RT‐qPCR results). In the dd‐PCR test, 11 of the 61 patients with CML (18.03%) tested positive and showed statistically significant difference (P < .01). In the follow‐up of 10 CML patients who were in MR4.5. All patients were loss of MR4.0, and 4 were tested positive by dd‐PCR 3 months earlier than by RT‐qPCR.
Conclusion
In contrast with RT‐qPCR, dd‐PCR is more sensitive, thus enabling accurate conversion of dd‐PCR results into internationally standard RT‐qPCR results by conversion equation, to achieve a deeper molecular biology‐based stratification of BCR/ABL(P210) MRD. It has some reference value to monitor disease progression in clinic.
Numerous outbreak investigations and case-control studies of campylobacteriosis have provided evidence that handling Campylobacter-contaminated chicken products is a high risk factor for infection ...and illness. In this study, the cross-contamination and transfer rates of Campylobacter jejuni from chicken to ready-to-eat food were determined in various food handling scenarios. Skinless raw chicken breasts were artificially contaminated with C. jejuni and diced on cutting boards of three different materials. Whether cold water, cold water with detergent, or hot water was used, statistically significant differences were found between the transfer rates of C. jejuni to unwashed and washed cutting boards or hands, respectively. When both kitchen knife and cutting board were reused after dicing the artificially contaminated chicken, the transfer rates of C. jejuni to cucumber cut on bamboo, wooden, and plastic cutting boards were 16.28, 12.82, and 5.32%, respectively. The transfer rates from chicken to bread, a large lift-up water faucet handle, and a small twist faucet handle via unwashed hands were 0.49, 4.64, and 3.14%, respectively. This research provides scientific evidence that various types of contaminated kitchenware and cook's hands are vital potential vehicles for the cross-contamination of Campylobacter from raw chicken to ready-to-eat food and emphasizes the importance of timely and proper cleaning to prevent cross-contamination during food handling; therefore, high-quality consumer education to reduce the risk of foodborne infection is urgent and necessary.
A genetic dereplication approach in combination with differential gene expression led to the discovery of three new sesquiterpenes, tricinoloniol acids (TRAs) A-C (1-3) and the known fusidilactone A ...(4) from T. hypoxylon. Comparative transcriptomic analysis and targeted deletion identified the biosynthetic route for TRAs. Our results demonstrate an alternative application of the genetic dereplication method for exploring the biosynthesis of cryptic secondary metabolites (SMs), which utilizes the coordinated expression of trichothecene (tri) and tra cluster genes.
RT-qPCR offers high sensitivity, for accurate interpretations of qPCR results however, normalisation using suitable reference genes is fundamental. Androgens can regulate transcriptional expression ...including reference gene expression in prostate cancer. In this study, we evaluated ten mRNA and six non-protein coding RNA reference genes in five prostate cell lines under varied dihydrotestosterone (DHT) treatments. We validated the effects of DHT-treatments using media containing charcoal-stripped serum prior to DHT stimulation on the test samples by Western blot experiments. Reference gene expression stability was analysed using three programs (geNorm, NormFinder and BestKeeper), and the recommended comprehensive ranking is provided. Our results reveal that ACTB and GAPDH, and miR-16 and miR-1228-3p are the most suitable mRNA and miRNA reference genes across all cell lines, respectively. Considering prostate cancer cell types, ACTB/GAPDH and ACTB/HPRT1 are the most suitable reference gene combinations for mRNA analysis, and miR-16/miR-1228-3p and RNU6-2/RNU43 for miRNA analysis in AR+, and AR- and normal cell lines, respectively. Comparison of relative target gene (PCA3 and miR-141) expression reveals different patterns depending on reference genes used for normalisation. To our knowledge, this is the first report on validation of reference genes under different DHT treatments in prostate cancer cells. This study provides insights for discovery of reliable DHT-regulated genes in prostate cells.
Degradation of sulfamethoxazole (SMX) by the white-rot fungus Phanerochaete chrysosporium was assessed. P. chrysosporium had a stronger tolerance for SMX in the concentration range of 10–30mgL−1. The ...degradation percentage of SMX in liquid medium was 53% after 24h and reached 74% after 10days when SMX was added at 10mgL−1. Crude laccase produced by P. chrysosporium was used to degrade SMX in vitro. The results showed that the removal of SMX obviously increased with laccase activity and reaction time increasing.
Display omitted
•Biodegradation was the main removal mechanism of sulfamethoxazole by Phanerochaete chrysosporium.•Laccase of P. chrysosporium plays a key role in sulfamethoxazole biodegradation.•The removal of sulfamethoxazole increased with laccase activity and reaction time increasing.
•The IC50 of phenol on Anammox was 678.2mgL−1 in short-term test.•The modified non-competitive inhibition model fit the test data well.•The evolution of Anammox performance under phenol inhibition ...was presented.•Phenol changed stoichiometric ratios and granule properties.•Measures for recovery of Anammox performance are provided.
The short- and long-term effects of phenol on anaerobic ammonium oxidation (Anammox) were evaluated. The short-term impact of phenol on Anammox activity was determined by a batch test, and an IC50 value of 678.2mgL−1 was calculated. Anammox granular sludge was equally seeded into two identical upflow anaerobic sludge blanket reactors (R0 and R1); synthetic wastewater without phenol was fed to R0 while with varied phenol was fed to R1 to study the long-term effects. The performance of R0 was stable, with a steadily rising nitrogen removal rate of 10.5–21.3kgNm−3day−1. However, the performance of R1 was significantly suppressed by an influent phenol concentration of 50mgL−1, and was recovered and stabilized by applying one or more control strategies. The phenol-mediated inhibition depressed the Anammox activity and biomass, and caused a change of stoichiometric ratios and granule characteristics.
Summary
Background
Alflutinib is a novel irreversible and highly selective third-generation EGFR inhibitor currently being developed for the treatment of non-small cell lung cancer patients with ...activating EGFR mutations and EGFR T790M drug-resistant mutation. Alflutinib is mainly metabolized via CYP3A4 to form its active metabolite AST5902. Both alflutinib and AST5902 contribute to the in vivo pharmacological activity. The aim of this study was to investigate the effects of rifampicin (a strong CYP3A4 inducer) on the pharmacokinetics of alflutinib and AST5902 in healthy volunteers, thus providing important information for drug-drug interaction evaluation and guiding clinical usage.
Methods
This study was designed as a single-center, open-label, and single-sequence trial over two periods. The volunteers received a single dose of 80 mg alflutinib on Day 1/22 and continuous doses of 0.6 g rifampicin on Day 15–30. Blood sampling was conducted on Day 1–10 and Day 22–31. The pharmacokinetics of alflutinib, AST5902, and the total active ingredients (alflutinib and AST5902) with or without rifampicin co-administration were respectively analyzed.
Results
Co-administration with rifampicin led to 86% and 60% decreases in alflutinib AUC
0-∞
and C
max
, respectively, as well as 17% decrease in AST5902 AUC
0-∞
and 1.09-fold increase in AST5902 C
max
. The total active ingredients (alflutinib and AST5902) exhibited 62% and 39% decreases in AUC
0-∞
and C
max
, respectively.
Conclusions
As a strong CYP3A4 inducer, rifampicin exerted significant effects on the pharmacokinetics of alflutinib and the total active ingredients (alflutinib and AST5902). The results suggested that concomitant strong CYP3A4 inducers should be avoided during alflutinib treatment. This trial was registered at
http://www.chinadrugtrials.org.cn
. The registration No. is CTR20191562, and the date of registration is 2019-09-12.
PDF
