Oridonin inhibits bladder cancer survival and immune escape by covalently targeting HK1.

BACKGROUND: Hexokinase I (HK1) is highly expressed in a variety of malignancies, regulates glycolytic pathway in cancer cells, and thus considered to be one of the promising molecular targets for cancer therapy. Nonetheless, the development of a specific inhibitor against HK1 remains elusive. PURPOSE: This study aims to elucidate the mechanism by which oridonin inhibits the proliferation and immune evasion of bladder cancer cells, specifically through the suppression of HK1. METHODS: To examine the mechanisms by which oridonin directly binds to cysteines of HK1 and inhibits bladder cancer growth, this study utilized a variety of methods. These included the Human Proteome Microarray, Streptavidin-agarose affinity assay, Biolayer Interferometry (BLI) ainding analysis, Mass Spectrometry, Cellular Thermal Shift Assay, Extracellular Acidification Rate measurement, and Xenotransplant mouse models. RESULTS: As indicated by our current findings, oridonin forms a covalent bond with Cys-813, located adjacently to glucose-binding domain of HK1. This suppresses the enzymatic activity of HK1, leading to an effective reduction of glycolysis, which triggers cell death via apoptosis in cells derived from human bladder cancer. Significantly, oridonin also inhibits lactate-induced PD-L1 expression in bladder cancer. Furthermore, pairing oridonin with a PD-L1 inhibitor amplifies the cytotoxicity of CD8+ T cells against bladder cancer. CONCLUSION: This research strongly suggests that oridonin serves as a covalent inhibitor of HK1. Moreover, it indicates that functional cysteine residue of HK1 could operate as viable targets for selective inhibition. Consequently, oridonin exhibits substantial potential for the evolution of anti-cancer agents targeting the potential therapeutic target HK1 via metabolism immunomodulation.

Mechanical wounds expedited starch degradation in the wound tissues of potato tubers.

Starch degradation occurs rapidly in stressed plants, but it is unclear how starch degradation occurs in potato tubers after they incur mechanical wounding. In this study, we found that wounding significantly upregulated the expression levels of StGWD, StAMY, StBAM, and StISA, and decreased the starch content of potato tubers. Meanwhile, wounding markedly upregulated the expression levels of StSUS, StBG, and StINV genes, and increased the content of sucrose, glucose, and fructose. Furthermore, wounding reduced the proportion of small starch granules and increase that of large as well as medium starch granules, in this way enhancing the average size distribution of starch. Initially, the hard surface layer of starch granules was removed by wounding, but the internal channels and other structures were only slightly affected. Taken together, the results show that wounding can accelerate starch degradation by promoting the accumulation of sucrose, glucose, and fructose, and the hydrolysis of starch granules in potato tubers.

AaCaM is required for infection structure differentiation and secondary metabolites in pear fungal pathogen Alternaria alternata.

AIMS: Calmodulin (CaM), acts as a kind of multifunctional Ca2+ sensing protein, which is ubiquitous in fungi, is highly conserved across eukaryotes and is involved in the regulation of a range of physiological processes, including morphogenesis, reproduction and secondary metabolites biosynthesis. Our aim was to understand the characteristics and functions of AaCaM in Alternaria alternata, the causal agent of pear black spot. METHODS AND RESULTS: A 450 bp cDNA sequence of AaCaM gene of A. alternata was cloned by the PCR homology method. Sequence analysis showed that this protein encoded by AaCaM was a stable hydrophilic protein and had a high similarity to Neurospora crassa (CAA50271.1) and other fungi. RT-qPCR analysis determined that AaCaM was differentially upregulated during infection structural differentiation of A. alternata both on hydrophobic and pear wax extract-coated surface, with a 3.37-fold upregulation during the hydrophobic induced appressorium formation period (6 h) and a 1.46-fold upregulation during the infection hyphae formation period (8 h) following pear wax induction. Pharmaceutical analysis showed that the CaM-specific inhibitor, trifluoperazine (TFP), inhibited spore germination and appressorium formation, and affected toxins and melanin biosynthesis in A. alternata. CONCLUSIONS: AaCaM plays an important role in regulating infection structure differentiation and secondary metabolism of A. alternata. SIGNIFICANCE AND IMPACT OF STUDY: Our study provides a theoretical basis for further in-depth investigation of the specific role of AaCaM in the calcium signalling pathway underlying hydrophobic and pear wax-induced infection structure differentiation and pathogenicity of A. alternata.

Corylin, a flavonoid derived from Psoralea Fructus, induces osteoblastic differentiation via estrogen and Wnt/ß-catenin signaling pathways.

Corylin is a naturally occurring flavonoid isolated from the fruit of Psoralea corylifolia L. (Fabaceae), which is a Chinese medicinal herb in treating osteoporosis. Although a variety of pharmacological activities of corylin have been reported, its osteogenic action and the underlying mechanism in bone development remain unclear. In the present study, the involvement of bone-specific genes in corylininduced differentiated osteoblasts was analyzed by RT-PCR, promoter-reporter assay, and Western blotting. In cultured osteoblasts, corylin-induced cell differentiation and mineralization, as well as increased the expressions of vital biological markers for osteogenesis, such as Runx2, Osterix, Col1, and ALP. Corylin was proposed to have dual pathways in triggering the osteoblastic differentiation. First, the osteogenic function of corylin acted through the activation of Wnt/ß-catenin signaling. The nuclear translocation of ß-catenin of cultured osteoblasts, as determined by flow cytometry and confocal microscopy, was triggered by applied corylin, and which was blocked by DKK-1, an inhibitor of Wnt/ß-catenin signaling. Second, the application of corylin-induced estrogenic response in a dose-dependent manner, and which was blocked by ICI 182 780, an antagonist of estrogen receptor. Furthermore, the activation of Runx2 promoter by corylin was abolished by both DKK-1 and ICI 182,780, indicating that the corylin exhibited its osteogenic effect via estrogen and Wnt/ß-catenin signaling pathways. In addition, corylin regulated the metabolic profiles, as well as the membrane potential of mitochondria, in cultured osteoblasts. Corylin also stimulated the osteogenesis in bone micromass derived from mesenchymal progenitor cells. This study demonstrated the osteogenic activities of corylin in osteoblasts and micromass, suggesting that corylin has the potential to be developed as a novel pro-osteogenic agent in targeting for the treatment of osteoblast-mediated osteoporosis.

Alteration of phenolic profiles and antioxidant capacities of common buckwheat and tartary buckwheat produced in China upon thermal processing.

BACKGROUND: Buckwheat products are receiving increasing attention because of their high nutritive values and significant health-promoting properties. In the present study, 15 buckwheat products grown in different parts of China were investigated. Representative common or tartary buckwheat samples were further subjected to soaking, roasting, microwave cooking, boiling and steaming treatments. Colorimetric analyses and high-performance liquid chromatography (HPLC) analyses were performed to determine the phenolic profiles and antioxidant capacities of the raw and thermally processed buckwheat samples, respectively. RESULTS: Tartary buckwheat exhibited a remarkably higher total phenolic content (TPC), total flavonoid content (TFC), 2-diphenyl-1-picryhydrazyl (DPPH) free radical scavenging activity and ferric reducing antioxidant power (FRAP) compared to common buckwheat, although there were no significant differences between their 2,2'-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS) free radical scavenging capacity. All thermal treatments, particularly microwave cooking, contributed to the greatest losses of phenolics and antioxidant capacities in the common buckwheat samples, whereas boiling and steaming usually resulted in the lowest losses. For the tartary buckwheat samples, all thermal treatments (except roasting), especially boiling and steaming, led to significant increases in TPC, TFC, DPPH free radical scavenging activity, FRAP and ABTS free radical scavenging capacity. However, HPLC analyses indicated that all thermal treatments, especially microwave cooking, gave rise to the greatest losses of the total content of 14 phenolic acids and three flavonoids, whereas boiling led to the lowest losses. CONCLUSION: Both steaming and boiling treatments are recommended when preparing common or tartary buckwheat food products because they can minimize thermal degradation or promote their phenolic compounds and antioxidant capacities to the greatest extent. © 2019 Society of Chemical Industry.

Comparative genomic and transcriptomic analyses of the Fuzhuan brick tea-fermentation fungus Aspergillus cristatus.

BACKGROUND: Aspergillus cristatus is the dominant fungus involved in the fermentation of Chinese Fuzhuan brick tea. Aspergillus cristatus is a homothallic fungus that undergoes a sexual stage without asexual conidiation when cultured in hypotonic medium. The asexual stage is induced by a high salt concentration, which completely inhibits sexual development. The taxon is therefore appropriate for investigating the mechanisms of asexual and sexual reproduction in fungi. In this study, de novo genome sequencing and analysis of transcriptomes during culture under high- and low-osmolarity conditions were performed. These analyses facilitated investigation of the evolution of mating-type genes, which determine the mode of sexual reproduction, in A. cristatus, the response of the high-osmolarity glycerol (HOG) pathway to osmotic stimulation, and the detection of mycotoxins and evaluation of the relationship with the location of the encoding genes. RESULTS: The A. cristatus genome comprised 27.9 Mb and included 68 scaffolds, from which 10,136 protein-coding gene models were predicted. A phylogenetic analysis suggested a considerable phylogenetic distance between A. cristatus and A. nidulans. Comparison of the mating-type gene loci among Aspergillus species indicated that the mode in A. cristatus differs from those in other Aspergillus species. The components of the HOG pathway were conserved in the genome of A. cristatus. Differential gene expression analysis in A. cristatus using RNA-Seq demonstrated that the expression of most genes in the HOG pathway was unaffected by osmotic pressure. No gene clusters associated with the production of carcinogens were detected. CONCLUSIONS: A model of the mating-type locus in A. cristatus is reported for the first time. Aspergillus cristatus has evolved various mechanisms to cope with high osmotic stress. As a fungus associated with Fuzhuan tea, it is considered to be safe under low- and high-osmolarity conditions.

In vitro antiviral effect of germacrone on feline calicivirus.

Feline calicivirus (FCV) often causes respiratory tract and oral disease in cats and is a highly contagious virus. Widespread vaccination does not prevent the spread of FCV. Furthermore, the low fidelity of the RNA-dependent RNA polymerase of FCV leads to the emergence of new variants, some of which show increased virulence. Currently, few effective anti-FCV drugs are available. Here, we found that germacrone, one of the main constituents of volatile oil from rhizoma curcuma, was able to effectively reduce the growth of FCV strain F9 in vitro. This compound exhibited a strong anti-FCV effect mainly in the early phase of the viral life cycle. The antiviral effect depended on the concentration of the drug. In addition, germacrone treatment had a significant inhibitory effect against two other reference strains, 2280 and Bolin, and resulted in a significant reduction in the replication of strains WZ-1 and HRB-SS, which were recently isolated in China. This is the first report of antiviral effects of germacrone against a calicivirus, and extensive in vivo research is needed to evaluate this drug as an antiviral therapeutic agent for FCV.

Bio-inspired synthesis yields a tricyclic indoline that selectively resensitizes methicillin-resistant Staphylococcus aureus (MRSA) to ß-lactam antibiotics.

The continuous emergence of resistant bacteria has become a major worldwide health threat. The current development of new antibacterials has lagged far behind. To discover reagents to fight against resistant bacteria, we initiated a chemical approach by synthesizing and screening a small molecule library, reminiscent of the polycyclic indole alkaloids. Indole alkaloids are a class of structurally diverse natural products, many of which were isolated from plants that have been used as traditional medicine for millennia. Specifically, we adapted an evolutionarily conserved biosynthetic strategy and developed a concise and unified diversity synthesis pathway. Using this pathway, we synthesized 120 polycyclic indolines that contain 26 distinct skeletons and a wide variety of functional groups. A tricyclic indoline, Of1, was discovered to selectively potentiate the activity of ß-lactam antibiotics in multidrug-resistant methicillin-resistant Staphylococcus aureus (MRSA), but not in methicillin-sensitive S. aureus. In addition, we found that Of1 itself does not have antiproliferative activity but can resensitize several MRSA strains to the ß-lactam antibiotics that are widely used in the clinic, such as an extended-spectrum ß-lactam antibiotic amoxicillin/clavulanic acid and a first-generation cephalosporin cefazolin. These data suggest that Of1 is a unique selective resistance-modifying agent for ß-lactam antibiotics, and it may be further developed to fight against resistant bacteria in the clinic.

[Infection progress of arbuscular mycorrhizae on tissue-cultured plantlets of Pinellia ternata].

OBJECTIVE: To study the Arbuscular mycorrhizal (AM) formation progress and infection characteristics between tissue culture plantlets of Pinellia ternata and Glomus mosseae. METHOD: The tissue culture plantlets of P. ternata were inoculated with G. mosseae, the formation of AM were sampled and observed with microscopy by staining. RESULT AND CONCLUSION: The hyphae of G. mosseae began to penetrate the root epidermis after 10 days of inoculation. Lots of intracellular hyphae formed in cortex cells at the 15th day. Arbuscules started to form and there were some hyphae on the root at the 20th day. At the 25th day, many arbuscules formed and most as Arum type. Some arbuscles started to disintegrate at the 30th day, and a few of vesicles occurred. Lots of spores formed after 35 days. At the 40th day, some vesicles began to decline. The hand section showed that the intercellular hyphae gradually formed in intercellular space, and the hyphae branched in cortex cells and occupied most cell lumen finally. It is expounded that P. ternata and G. mosseae could recognize each other quickly and form a symbiont system.

Effects of source and level of magnesium on catalase activity and its gene expression in livers of broiler chickens.

The effects of dietary supplemental magnesium oxide (MgO), magnesium-L-aspartate (MgAsp) and monomagnesium-di-L-aspartate (MgdiAsp) on hepatic catalase (CAT) activity and its mRNA expression were investigated. A total of 360 one-day-old male Abor Acre broiler chickens were allocated to ten treatments, i.e. control plus 9 treatments from 3 x 3 factorial arrangement (Mg source, Mg level), each treatment with six replicates of 6 chickens. The birds were fed with the basal diet alone or supplemented with magnesium (Mg) at 0.9, 1.8, 2.7 g/kg of the diet from MgO, MgAsp or MgdiAsp. Results showed that hepatic Mg concentration increased quadratically as MgO or MgAsp supplementation increased (p < 0.01). Hepatic CAT activity increased linearly in birds fed with MgAsp or MgdiAsp (p < 0.01) and quadratically in birds fed with MgO (p < 0.05) as dietary Mg supplementation level increased. Hepatic CAT mRNA was linearly correlated with the dietary Mg supplementation level (p < 0.01). There were positive correlations among hepatic CAT activity, its mRNA expression level and hepatic Mg concentration (p < 0.01). No effect of Mg2+ on the purified CAT activity was detected in vitro enzymatic reaction system (p > 0.05). Supplemental MgAsp or MgdiAsp was more efficient to increase hepatic Mg concentrations, enhance hepatic CAT activity and its mRNA expression than MgO (p < 0.01). It can be concluded that dietary Mg supplementation could increase hepatic Mg concentration, enhance CAT mRNA expression and consequently enhance CAT activity, and the organic Mg (MgAsp or MgdiAsp) is much more efficient than the inorganic form (MgO).

[Basis of art phonetics in biomedical engineering].

Art phonetics' medicine, a new branch of traditional medicine, has not been developed perfectly, especially in the aspects of objective and scientific study. In this paper, the acoustical and anatiomical basis of art phonetics in viewpoint of biomedical engineering is explored, and then our work of quantitative measurement and analysis of art phonetic is introduced. The experiment data show further that quantitative measurement and analysis plays an important role in art phonetic medicine.