Transcription factor GlbHLH regulates hyphal growth, stress resistance, and polysaccharide biosynthesis in Ganoderma lucidum.

Basic helix-loop-helix (bHLH) transcription factors (TFs) participate in many physiological and cellular processes in eukaryotes. However, their functions remain unclear in the macro basidiomycete Ganoderma lucidum (G. lucidum). In this study, a gene encoding bHLH TF, GlbHLH, was identified in G. lucidum. The knockdown of GlbHLH by RNA interference reduced hyphal growth, hyphal branching, and resistant to osmotic, oxidative, and cell wall stress. The content of cell wall components ß-1,3 glucan and chitin and the expression of their synthesis genes were decreased in the GlbHLH knockdown strains. The knockdown of GlbHLH led to an increase of intracellular reactive oxygen species by decreasing the enzyme activity and gene expression of antioxidant enzymes. Furthermore, the production of intracellular polysaccharides and extracellular polysaccharides was greatly decreased in the GlbHLH mutants. These results suggested that GlbHLH is involved in hyphal growth, stress response, and polysaccharide biosynthesis in G. lucidum.

Protective Effects of Herba Houttuyniae Aqueous Extract against OVA-Induced Airway Hyperresponsiveness and Inflammation in Asthmatic Mice.

Herba Houttuyniae is the well-knownfood-medicine herb with the special taste and smell. It is also widely used in south China for prevention of various chronic pulmonary inflammatory diseases including asthma. However, the active ingredients and therapeutic mechanism of this herb remain obscure. In this study, network pharmacology technology was employed to investigate the effects of Herba Houttuyniae aqueous extract (HHAE) on OVA-induced airway hyperresponsiveness and inflammation. The results showed that six compounds (isoramanone, kaempferol, 1-methyl-2-nonacosyl-4-quinolone, C09747, spinasterol, and quercetin) were found to be mainly responsible for the therapeutic effects of the herb, which totally regulated the expressions of 168 asthma-related proteins. All those targets involved in the signal transduction of the prolactin signaling pathway, central carbon metabolism in cancer, EGFR tyrosine kinase inhibitor resistance, endocrine resistance, and VEGF signaling pathway. The in vivo experiment also revealed that orally administrated with HHAE alleviated airway hyperresponsiveness and inflammation in OVA-induced asthmatic mice. It significantly decreased the counts of neutrophils, eosinophils, and lymphocytes as well as the levels of IL-1ß, IL-4, IL-6, and IL-13 in BALF of asthmatic mice. Mechanically, HHAE downregulated both the mRNA and protein expressions of p38 MAPK, PI3K, AKT, and VEGF in the lung tissues of asthmatic mice. Therefore, HHAE improved OVA-induced airway hyperresponsiveness and inflammation in mice and could be a potential supplement for asthma treatment.

Long non-coding RNA MINCR regulates the growth and metastasis of human osteosarcoma cells via Wnt/ß-catenin signaling pathway.

There is concrete evidence that lncRNA-MINCR is involved in tumorigenesis of a number of human cancers through modulation of Wnt/ß-catenin signaling pathway. However, the characterization of regulatory role of lncRNA-MINCR has not been worked out in osteosarcoma yet. The present study was undertaken to explore the role of lncRNA-MINCR in human osteosarcoma. The osteosarcoma tissues and cell lines were found to exhibit significant (P<0.05) over-expression of lncRNA-MINCR. Silencing of lncRNA-MINCR in osteosarcoma cells suppressed their cell viability through the induction of apoptosis. The Saos-2 osteosarcoma cells exhibited significant (P<0.05) decline in migration and invasion rate together with inhibition of EMT under transcriptional knockdown of lncRNA-MINCR. Western blot analysis revealed that lncRNA-MINCR operated through Wnt/ß-catenin signaling pathway to control the growth and metastasis of osteosarcoma cells. In vivo mice tumorigenesis was significantly (P<0.05) restricted under lncRNA-MINCR repression. The study clearly indicated that lncRNA-MINCR exhibits crucial growth regulatory role in osteosarcoma together with its ability to control the metastasis of cancer cells through Wnt/ß-catenin signal.

Qualitative and quantitative analysis of sesquiterpene lactones in Centipeda minima by UPLC-Orbitrap-MS & UPLC-QQQ-MS.

An ultra-high performance liquid chromatography coupled with high-resolution Orbitrap mass spectrometry (UPLC-Orbitrap-MS) method has been used to identify sesquiterpene lactones in the methanolic extract of Centipeda minima. Fifteen sesquiterpene lactones were tentatively identified based on retention time and accurate mass of external standards or exact accurate mass searching (within 2 ppm) by comparison of some previous isolated sesquiterpene lactones. Meanwhile, a rapid, sensitive, precise, and reliable ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-QQQ-MS) method has been developed to evaluate the quality of Centipeda minima through a simultaneous determination of five sesquiterpene lactones, namely brevilin A, arnicolide C, arnicolide D, microhelenin C, minimolide F. Chromatographic separation was achieved on a Waters Acquilty UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 µm) with a mobile phase consisting of a) 0.1% formic acid and b) a mixture of acetonitrile and methanol 50:50 v/v under an isocratic elution (42:58) manner. Positive electrospray ionization mode with multiple reaction monitoring was applied for the detection of the five sesquiterpene lactones. Method validation for linearity, accuracy and precision was also carried out. Finally, the method was successfully used for the analysis of 10 batches of Centipeda minima samples collected in China. Brevilin A and arnicolide D were the dominant sesquiterpene lactones in Centipeda minima and could be proposed as suitable markers for the quality control of Centipeda minima.

Inhibition of proliferation and gene expression regulation by (-)-epigallocatechin-3-gallate in human synovial sarcoma cells.

Synovial sarcoma is an aggressive soft-tissue malignancy with poor prognosis and lack of response to conventional cytotoxic chemotherapy. The regulatory mechanisms for the rapid proliferation of synovial sarcoma cells and the particular aggressiveness of this sarcoma remain poorly understood. In this study, we investigate the effect of epigallocatechin-3-gallate (EGCG) on growth and apoptosis of chondrosarcoma cells. The MTT assay and DAPI staining indicated that EGCG effectively inhibited cellular proliferation and induces apoptosis of the synovial sarcoma cells and induced apoptosis as confirmed by flow cytometry. Furthermore, Bcl-2 and Mcl-1 levels significantly decreased, Bax levels significantly increased, whereas expression levels of the proteins Bcl-XL were unchanged in response to EGCG treatment in SW982. In conclusion, our findings demonstrate that EGCG is effective for growth inhibition of synovial sarcoma cell lines in vitro and suggest that EGCG may be a new therapeutic option for patients with synovial sarcoma.