Exploring the anti-glioma mechanism of the active components of Cortex Periplocae based on network pharmacology and iTRAQ proteomics in vitro.

BACKGROUND: The active components of Cortex Periplocae (CP) exert antitumor properties in many cancers. However, little is known about their effects on glioma or the related underlying mechanisms. OBJECTIVES: The study investigated the underlying mechanism of CP in treating glioma. MATERIAL AND METHODS: The U251 and TG905 cells were treated with an ethanol extract from CP. Cell proliferation was detected using Cell Counting Kit-8 (CCK-8) and a colony formation assay. The flow cytometric analysis was applied to explore the induction of cell cycle arrest and apoptosis. The expression levels of cell cycleand apoptosis-associated proteins were measured with western blot. A network pharmacology method was performed to predict the potential mechanism underlying the effects of the active components of CP on glioma. Then, isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics analysis was used to verify the differentially expressed proteins and pathways in order to reveal the underlying mechanisms. Furthermore, to determine the iTRAQ results, 6 candidate proteins were chosen for quantification using parallel reaction monitoring (PRM). RESULTS: The CP extract inhibited the proliferation of U251 and TG905 cells and induced cell cycle arrest and apoptosis. There are 16 active compounds of CP. The antitumor mechanism of CP may be related to the apoptosis pathway, p53 signaling pathway, PI3K-AKT pathway, or transcriptional misregulation in cancer pathway. Six proteins (HSP90AB1, TOP2A, ATP1A1, TGFß1, ATP1B1, and TYMS) were determined to be key factors involved in regulating CP in glioma. CONCLUSIONS: Our research revealed the underlying mechanism of CP in treating glioma using integrated network pharmacology and iTRAQ-based quantitative proteomics technology.

Stimulating mechanism of corn oil on biomass and polysaccharide production of Pleurotus tuber-regium mycelium.

Hyperbranched polysaccharides (HBPSs) are the main components in cell wall and exopolysaccharide (EPS) of Pleurotus tuber-regium. To enhance the yield of these macromolecules, corn oil at 4% addition exhibited the best effect for production of mycelial biomass at 20.49 g/L and EPS at 0.59 g/L, which was 2.56 folds and 1.90 folds of the control, respectively. The treated hyphae were much thicker with smooth surface, while its cell wall content (43.81 ± 0.02%) was 1.96 times of the control (22.34 ± 0.01%). Moreover, a large number of lipid droplets could be visualized under the view of confocal laser scanning microscopy (CLSM). RNA-seq analysis revealed that corn oil could enter the cells and result in the up-regulation of genes on cell morphology and membrane permeability, as well as the down-regulation on expression level of polysaccharide hydrolase and genes involved in the MAPK pathway, all of which probably contribute to the increase of polysaccharides production.

[Chromosomal studies of germ cells in diploid and polyploid fish produced by distant crossing].

By observing the chromosomal spreads of germ cells and tissue sections, we studied the chromosomal spreads of germ cells in diploid and polyploid fish produced by distant crossing. The samples covered the second generation of hybrids of red crucian carp (Carassius auratus red var.) (female) x common carp (Cyprinus carpio) (male) (2n = 100) (F2), allotetraploid hybrids of red crucian carp (female) x common carp (male)(4n = 200), triploid hybrids of gold fish (female)x allotetraploid (male) (3n = 150), the second generation of the gynogenetic progeny of allotetraploid hybrids (G2) (2n = 100), and the common carp (2n = 100) used as a control. The results demonstrated that chromosomal number of spermotogonia in common carp was equal to that of their somatic cell (2n = 100), while the chromosomal number of germ cells in diploid hybrid fish and polyploid hybrid fish had doubled obviously, and the frequent of chromosomal doubling in spermotogonia of F2 appeared especially high, accounting for 21.6% of all examined chromosomal spreads, which provided directly cytological evidence for the production of unreduced diploid gametes in F2 and also indicated that distant crossing was an important factor to lead to chromosomal doubling in germ cell. This research had important significance in studies on the formation of polyploidy fish and fish breeding.