Virtual screening-molecular docking-activity evaluation of Ailanthus altissima (Mill.) swingle bark in the treatment of ulcerative colitis.

BACKGROUND: The dried bark of Ailanthus altissima (Mill.) Swingle is widely used in traditional Chinese medicine for the treatment of ulcerative colitis. The objective of this study was to explore the therapeutic basis of the dried bark of Ailanthus altissima (Mill.) Swingle for the treatment of ulcerative colitis based on Virtual Screening-Molecular Docking-Activity Evaluation technology. METHODS: By searching the Traditional Chinese Medicine Systems Pharmacology TCMSP Database and Analysis Platform, 89 compounds were obtained from the chemical components of the dried bark of Ailanthus altissima (Mill.) Swingle. Then, after preliminarily screening the compounds based on Lipinski's rule of five and other relevant conditions, the AutoDock Vina molecular docking software was used to evaluate the affinity of the compounds to ulcerative colitis-related target proteins and their binding modes through use of the scoring function to identify the best candidate compounds. Further verification of the compound's properties was achieved through in vitro experiments. RESULTS: Twenty-two compounds obtained from the secondary screening were molecularly docked with ulcerative colitis-related target proteins (IL-1R, TLR, EGFR, TGFR, and Wnt) using AutoDock Vina. The free energies of the highest scoring compounds binding to the active cavity of human IL-1R, TLR, EGFR, TGFR, and Wnt proteins were - 8.7, - 8.0, - 9.2, - 7.7, and - 8.5 kcal/mol, respectively. The potential compounds, dehydrocrebanine, ailanthone, and kaempferol, were obtained through scoring function and docking mode analysis. Furthermore, the potential compound ailanthone (1, 3, and 10 µM) was found to have no significant effect on cell proliferation, though at 10 µM it reduced the level of pro-inflammatory factors caused by lipopolysaccharide. CONCLUSION: Among the active components of the dried bark of Ailanthus altissima (Mill.) Swingle, ailanthone plays a major role in its anti-inflammatory properties. The present study shows that ailanthone has advantages in cell proliferation and in inhibiting of inflammation, but further animal research is needed to confirm its pharmaceutical potential.

Identifying the Genes Regulated by AtWRKY6 Using Comparative Transcript and Proteomic Analysis under Phosphorus Deficiency.

Phosphorus (P) is an important mineral nutrient for plant growth and development. Overexpressing AtWRKY6 (35S:WRKY6-9) was more sensitive and wrky6 (wrky6-1) was more resistant under low Pi conditions. To better understand the function of AtWRKY6 under low phosphate stress conditions, we applied two-dimensional gel electrophoresis (2-DE) to analyse differentially expressed proteins in the shoots and roots between wild type, 35S:WRKY6-9 and wrky6-1 after phosphorus deficiency treatment for three days. The results showed 88 differentially abundant protein spots, which were identified between the shoots and roots of 35S:WRKY6-9 and wrky6-1 plants. In addition, 59 differentially expressed proteins were identified in the leaves and roots of 35S:WRKY6-9 plants. After analysis, 9 genes with W-box elements in their promoter sequences were identified in the leaves, while 6 genes with W-box elements in their promoter sequences were identified in the roots. A total of 8 genes were identified as potential target genes according to the quantitative PCR (QPCR) and two dimension difference gel electrophoresis, (2D-DIGE) results, including ATP synthase, gln synthetase, nitrilase, 14-3-3 protein, carbonic anhydrases 2, and tryptophan synthase. These results provide important information concerning the AtWRKY6 regulation network and reveal potential vital target genes of AtWRKY6 under low phosphorus stress. two dimension difference gel electrophoresis, 2D-DIGE.

Biological phosphorus removal in anoxic-aerobic sequencing batch reactor with starch as sole carbon source.

In traditional biological phosphorus removal (BPR), phosphorus release in anaerobic stage is the prerequisite of phosphorus excessive uptake in aerobic conditions. Moreover, when low molecular weight of the organic substance such as volatile fatty acids (VFAs) is scarce in bulk liquid or anaerobic condition does not exist, phosphate accumulating organisms (PAOs) have difficulty removing phosphorus. However, in this work, phosphorus removal in two anoxic-aerobic sequencing batch reactors (SBRs) was observed when starch was supplied as a sole carbon source. The relations of the BPR with idle period were investigated in the two identical SBRs; the idle times were set to 0.5 hr (R1) and 4 hr (R2), respectively. Results of the study showed that, in the two SBRs, phosphorus concentrations of 0.26-3.11 mg/L in effluent were obtained after aeration when phosphorus concentration in influent was about 8 mg/L. Moreover, lower accumulations/transformations of polyhydroxyalkanoates (PHAs) and higher transformation of glycogen occurred in the SBRs, indicating that glycogen was the main energy source that was different from the traditional mechanism of BPR. Under the different idle time, the phosphorus removal was a little different. In R2, which had a longer idle period, phosphorus release was very obvious just as occurs in a anaerobic-aerobic regime, but there was a special phenomenon of chemical oxygen demand increase, while VFAs had no notable change. It is speculated that PAOs can assimilate organic compounds in the mixed liquor, which were generated from glycolysis by fermentative organisms, coupled with phosphorus release. In R1, which had a very short idle period, anaerobic condition did not exist; phosphorus removal rate reached 63%. It is implied that a new metabolic pathway can occur even without anaerobic phosphorus release when starch is supplied as the sole carbon source.

Phosphorus metabolism and population dynamics in a biological phosphate-removal system with simultaneous anaerobic phosphate stripping.

In this study, the metabolism of phosphorus and changes in population dynamics were investigated via simultaneous chemical stripping in sidestream in an acetate-fed sequencing batch reactor. The synthesized intracellular polyphosphate (poly-P) by polyphosphate-accumulating organisms (PAOs) gradually decreased when the biomass was subjected to 83 d of P stripping. Initially, the P removal efficiency of the system improved from 94.3% to 96.9%. Thereafter, a relatively high level of P in effluent was observed, during which time the stoichiometric ratios of Prelease/HAcuptake decreased, Glycogendegraded/HAcuptake and poly-ß-hydroxyvalerate/PHA increased. The results revealed that a metabolic shift from polyphosphate-accumulating metabolism to glycogen-accumulating metabolism. Correspondingly, PAOs declined to less than 1% of the population, glycogen-accumulating organisms proliferated to almost 20% instead. The results of PCR­DGGE also indicated that the microbial community structure considerably changed in response to the gradually decreasing poly-P content. These findings imply that intracellular poly-P level is important for the stable of P removal system. Furthermore, it suggests that it is not a stable and effective way for P recycling from anaerobic stage of the biological P removal system in sidestream.

Comparative analysis of Pu-erh and Fuzhuan teas by fully automatic headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry and chemometric methods.

Thirteen Pu-erh teas and 13 Fuzhuan teas obtained from two different production areas in China were profiled using fully automatic headspace solid-phase microextraction (HS-SPME)/gas chromatography-mass spectrometry (GC-MS) coupled with chemometric methods. A total of 93 aroma components were identified in 26 dark teas; among them, methoxyphenolic compounds (31.77%) were the most abundant components in Pu-erh teas, whereas ketone compounds were the most abundant components (25.42%) in Fuzhuan teas. Cluster analysis (CA) and principal component analysis (PCA) showed that these two types of dark teas could be clearly distinguished according to their chemical characteristics. This study suggested that the proposed strategy could provide a feasible and rapid technique to differentiate dark teas with similar morphological characteristics from different production areas by their volatile composition and relative content.