Optimization of the extraction process and metabonomics analysis of uric acid-reducing active substances from Gymnadenia R.Br. and its protective effect on hyperuricemia zebrafish

BackgroundAs Gymnadenia R.Br. (Gym) has an obvious uric acid-lowering effect, but its specific bioactive substances and mechanism are still unclear. The key metabolites and pathways used by Gym to reduce uric acid (UA) were identify.MethodsAn optimized extraction process for urate-lowering active substances from Gym was firstly been carried out based on the xanthine oxidase (XOD) inhibition model in vitro; then, the Ultra-high-performance liquid chromatography and Q-Exactive mass spectrometry (UHPLC-QE-MS) based on non-targeted metabolomics analysis of Traditional Chinese Medicine were performed for comparison of Gym with ethanol concentration of 95% (low extraction rate but high XOD inhibition rate) and 75% (high extraction rate but low XOD inhibition rate), respectively; finally, the protective effect of ethanolic extract of Gym on zebrafish with Hyperuricemia (referred to as HUA zebrafish) was explored.ResultsWe found that the inhibition rate of Gym extract with 95% ethanol concentration on XOD was 84.02%, and the extraction rate was 4.32%. Interestingly, when the other conditions were the same, the XOD inhibition rate of the Gym extract with 75% ethanol concentration was 76.84%, and the extraction rate was 14.68%. A total of 539 metabolites were identified, among them, 162 different metabolites were screened, of which 123 were up-regulated and 39 were down-regulated. Besides significantly reducing the contents of UA, BUN, CRE, ROS, MDA, and XOD activity in HUA zebrafish by Gym and acutely reduce the activity of SOD.ConclusionAlong with the flavonoids, polyphenols, alkaloids, terpenoids, and phenylpropanoids, the ethanolic extract of Gym may be related to reduce the UA level of Gym

Linearization Method of Nonlinear Magnetic Levitation System

Linearized model of the system is often used in control design. It is generally believed that we can obtain the linearized model as long as the Taylor expansion method is used for the nonlinear model. This paper points out that the Taylor expansion method is only applicable to the linearization of the original nonlinear function. If the Taylor expansion is used for the derived nonlinear equation, wrong results are often obtained. Taking the linearization model of the maglev system as an example, it is shown that the linearization should be carried out with the process of equation derivation. The model is verified by nonlinear system simulation in Simulink. The method in this paper is helpful to write the linearized equation of the control system correctly

Design of Stable Controller for Flexible Solar Panel by H∞ Loop-Shaping Method

Flexible solar panels play an essential role in the field of aerospace. However, many difficulties appear in the control design due to the existence of a weakly damped resonance module. The design for flexible systems often causes an unstable controller so that the systems after design still have trouble in putting into practice. We adopt H∞ loop-shaping design and put forward a directive method for selecting the weighting function. The simulation results indicate that system bandwidth is optimized based on the stable controller. In this way, the control precision and response speed of the system are improved. In the meantime, the system is easy to put into use

H∞ Observer Based on Descriptor Systems Applied to Estimate the State of Charge

This paper proposes an H∞ observer based on descriptor systems to estimate the state of charge (SOC). The battery’s open-current voltage is chosen as a generalized state variable, thereby avoiding the artificial derivative calculation of the algebraic equation for the SOC. Furthermore, the observer’s dynamic performance is saved. To decrease the impacts of the uncertain noise and parameter perturbations, nonlinear H∞ theory is implemented to design the observer. The sufficient conditions for the H∞ observer to guarantee the disturbance suppression performance index are given and proved by the Lyapunov stability theory. This paper systematically gives the design steps of battery SOC H∞ observers. The simulation results highlight the accuracy, transient performance, and robustness of the presented method

The Strica Homolog AaCASPS16 Is Involved in Apoptosis in the Yellow Fever Vector, Aedes albopictus.

Caspases are a family of cysteine proteases playing essential roles during apoptosis. Seven caspases identified in Drosophila were Dronc, Dredd, Strica, Dcp-1, Decay, Drice and Damm. Among them, Strica is an insect-specific caspase containing a long serine- and threonine- rich prodomain, of which function is not yet well studied. Here we identified a homolog of strica from Aedes albopictus, named as Aacasps16. Aacasps16 encoded a protein containing a putative serine- and threonine-rich prodomain and a well conserved caspase catalytic domain. AaCASPS16 shared high identity with dipteran insects Strica homologs. Alignment showed that the closest relative of AaCASPS16 was Aedes aegypti AeCASPS16. The expression profiles of Aacasps16 during developmental and adult stages were analyzed. Purified recombinant AaCASPS16 exhibited the highest caspase activity to WEHD, which is the substrate preferred by human caspase-9. AaCASPS16 induced apoptosis when over-expressed in C6/36 cells. AaCASPS16 was processed during apoptosis induced by actinomycin D and ultraviolet irradiation treatment, whereas partial silencing of Aacasps16 reduced actinomycin D- and ultraviolet irradiation-triggered apoptosis in C6/36 cells. Taken together, our study identified AaCASPS16 as a novel apoptotic caspase in Aedes albopictus

The sequence of AaCASPS16.

<p>Predicted amino acid sequence of AaCASPS16 was shown in alignment with Strica homologs from <i>Aedes aegypti</i> (AeCASPS15, AeCASPS16, AeCASPS17 and AeCASPS21) and the caspases from <i>Drosophila melanogaster</i> (Dronc, Dredd, Drice, Decay, Dcp-1, Strica and Damm). The amino acid residues identical among 12 caspases are indicated by white letters within black boxes, the amino acid residues identical among 9 caspases are indicated by black letters within dark gray boxes, the amino acid residues identical among 6 caspases are indicated by black letters within medium gray boxes, and the amino acid residues identical among 3 caspases are indicated by black letters within light gray boxes. The alignment was performed using DNAMAN 7.0. Secondary structures were predicted using JPred3. Underline: catalytic center, black arrow: predicted cleavage site.</p

AaCASPS16 was processed in apoptosis triggered by UV and Act D treatment.

<p>C6/36 cells were treated with Act D (1.0 μg/mL) or UV treatment (200 μJ/cm²) separately and at 24 h post treatment, cells were subjected to the following analyses: <b>(A)</b> Cell pictures were taken under microscope (Scale bar indicated 50 μm). <b>(B)</b> Cell lysates were prepared and incubated with Ac-DEVD-AFC and subjected to caspase activity assay. Caspase activity was indicated as the changes in relative fluorescence units (RFU) per minute. <b>(C)</b> Cell lysates were analyzed by immunoblotting using antibody against AaCASPS16 and β-actin. A short vertical black line was used to indicate where lanes were removed and separate parts of the same Western blot image were joined together. The data in (B) were presented with the SD from three independent experiments, and statistical significance was calculated by <i>t</i> test, **<i>P</i> < 0.01.</p

Transient expression of AaCASPS16 induced apoptosis in C6/36 cells.

<p>Plasmids expressing C-terminally Flag-tagged AaCASPS16-WT, AaCASPS16-C300A and GFP were transfected into C6/36 cells separately. Caspase inhibitor z-VAD-FMK was added at 2 h before transfection of pIE-AaCASPS16, and proteasome inhibitor MG132 was added at 8 h before cells were harvested. Mock treated cells, GFP expressed C6/36 cells, and AaCASPS16-C300A expressed C6/36 cells were used as controls. At 24 h post transfection, cells were subjected to the following analyses: <b>(A)</b> Photographs of cells were taken under microscope (Scale bar indicated 50 μm). <b>(B)</b> Cell lysates were prepared and were incubated with Ac-DEVD-AFC and subjected to caspase activity assay. Caspase activity was indicated as the changes in relative fluorescence units (RFU) per minute. <b>(C)</b> Cell lysates were subjected to immunoblotting analysis using antibody against Flag and β-tubulin. The data in (B) were presented with the SD from three independent experiments, and statistical significance was calculated by <i>t</i> test, ** <i>P</i> < 0.01. NS: not significant.</p

Data_Sheet_1_Optimization of the extraction process and metabonomics analysis of uric acid-reducing active substances from Gymnadenia R.Br. and its protective effect on hyperuricemia zebrafish.docx

BackgroundAs Gymnadenia R.Br. (Gym) has an obvious uric acid-lowering effect, but its specific bioactive substances and mechanism are still unclear. The key metabolites and pathways used by Gym to reduce uric acid (UA) were identify.MethodsAn optimized extraction process for urate-lowering active substances from Gym was firstly been carried out based on the xanthine oxidase (XOD) inhibition model in vitro; then, the Ultra-high-performance liquid chromatography and Q-Exactive mass spectrometry (UHPLC-QE-MS) based on non-targeted metabolomics analysis of Traditional Chinese Medicine were performed for comparison of Gym with ethanol concentration of 95% (low extraction rate but high XOD inhibition rate) and 75% (high extraction rate but low XOD inhibition rate), respectively; finally, the protective effect of ethanolic extract of Gym on zebrafish with Hyperuricemia (referred to as HUA zebrafish) was explored.ResultsWe found that the inhibition rate of Gym extract with 95% ethanol concentration on XOD was 84.02%, and the extraction rate was 4.32%. Interestingly, when the other conditions were the same, the XOD inhibition rate of the Gym extract with 75% ethanol concentration was 76.84%, and the extraction rate was 14.68%. A total of 539 metabolites were identified, among them, 162 different metabolites were screened, of which 123 were up-regulated and 39 were down-regulated. Besides significantly reducing the contents of UA, BUN, CRE, ROS, MDA, and XOD activity in HUA zebrafish by Gym and acutely reduce the activity of SOD.ConclusionAlong with the flavonoids, polyphenols, alkaloids, terpenoids, and phenylpropanoids, the ethanolic extract of Gym may be related to reduce the UA level of Gym.</p

Expression profile of <i>Aacasps16</i> in developmental and adult stages.

<p>Total RNAs were prepared from 1st to 4th instar larvae, pupae, female and male adults and subjected to qRT-PCR analysis. The vertical axis represents the relative expression of <i>Aacasps16</i> in different developmental stages or different genders. Pupae samples were designated as the standard and set to 1. The data were presented with the SD from three independent experiments. ND: not detected.</p