Characterization of anti-leukemia components from Indigo naturalis using comprehensive two-dimensional K562/cell membrane chromatography and in silico target identification.

Traditional Chinese Medicine (TCM) has been developed for thousands of years and has formed an integrated theoretical system based on a large amount of clinical practice. However, essential ingredients in TCM herbs have not been fully identified, and their precise mechanisms and targets are not elucidated. In this study, a new strategy combining comprehensive two-dimensional K562/cell membrane chromatographic system and in silico target identification was established to characterize active components from Indigo naturalis, a famous TCM herb that has been widely used for the treatment of leukemia in China, and their targets. Three active components, indirubin, tryptanthrin and isorhamnetin, were successfully characterized and their anti-leukemia effects were validated by cell viability and cell apoptosis assays. Isorhamnetin, with undefined cancer related targets, was selected for in silico target identification. Proto-oncogene tyrosine-protein kinase (Src) was identified as its membrane target and the dissociation constant (Kd) between Src and isorhamnetin was 3.81 μM. Furthermore, anti-leukemia effects of isorhamnetin were mediated by Src through inducing G2/M cell cycle arrest. The results demonstrated that the integrated strategy could efficiently characterize active components in TCM and their targets, which may bring a new light for a better understanding of the complex mechanism of herbal medicines

4-Methyl-N-(9-methyl-9-aza­bicyclo­[3.3.1]non-3-yl)benzamide

The asymmetric unit of the title compound, C17H24N2O, contains three independent mol­ecules. In the crystal, mol­ecules are linked by weak N—H⋯O hydrogen bonds into chains parallel to the c axis

Metabonomic Profiles Delineate the Effect of Traditional Chinese Medicine Sini Decoction on Myocardial Infarction in Rats

Background: In spite of great advances in target-oriented Western medicine for treating myocardial infarction (MI), it is still a leading cause of death in a worldwide epidemic. In contrast to Western medicine, Traditional Chinese medicine (TCM) uses a holistic and synergistic approach to restore the balance of Yin-Yang of body energy so the body’s normal function can be restored. Sini decoction (SND) is a well-known formula of TCM which has been used to treat MI for many years. However, its holistic activity evaluation and mechanistic understanding are still lacking due to its complex components. Methodology/Principal Findings: A urinary metabonomic method based on nuclear magnetic resonance and ultra highperformance liquid chromatography coupled to mass spectrometry was developed to characterize MI-related metabolic profiles and delineate the effect of SND on MI. With Elastic Net for classification and selection of biomarkers, nineteen potential biomarkers in rat urine were screened out, primarily related to myocardial energy metabolism, including the glycolysis, citrate cycle, amino acid metabolism, purine metabolism and pyrimidine metabolism. With the altered metabolism pathways as possible drug targets, we systematically analyze the therapeutic effect of SND, which demonstrated that SND administration could provide satisfactory effect on MI through partially regulating the perturbed myocardial energy metabolism. Conclusions/Significance: Our results showed that metabonomic approach offers a useful tool to identify MI-relate

Potential Biomarkers in Mouse Myocardium of Doxorubicin-Induced Cardiomyopathy: A Metabonomic Method and Its Application

BACKGROUND: Doxorubicin (DOX) is one of the most potent antitumor agents available; however, its clinical use is limited because of the risk of severe cardiotoxicity. Though numerous studies have ascribed DOX cardiomyopathy to specific cellular pathways, the precise mechanism remains obscure. Sini decoction (SND) is a well-known formula of Traditional Chinese Medicine (TCM) and is considered as efficient agents against DOX-induced cardiomyopathy. However, its action mechanisms are not well known due to its complex components. METHODOLOGY/PRINCIPAL FINDINGS: A tissue-targeted metabonomic method using gas chromatography-mass spectrometry was developed to characterize the metabolic profile of DOX-induced cardiomyopathy in mice. With Elastic Net for classification and selection of biomarkers, twenty-four metabolites corresponding to DOX-induced cardiomyopathy were screened out, primarily involving glycolysis, lipid metabolism, citrate cycle, and some amino acids metabolism. With these altered metabolic pathways as possible drug targets, we systematically analyzed the protective effect of TCM SND, which showed that SND administration could provide satisfactory effect on DOX-induced cardiomyopathy through partially regulating the perturbed metabolic pathways. CONCLUSIONS/SIGNIFICANCE: The results of the present study not only gave rise to a systematic view of the development of DOX-induced cardiomyopathy but also provided the theoretical basis to prevent or modify expected damage

Theoretical Analysis of Double Logistic Distributed Activation Energy Model for Thermal Decomposition Kinetics of Solid Fuels

The distributed activation energy model (DAEM) has been widely used to analyze the thermal decomposition of solid fuels such as lignocellulosic biomass and its components, coal, microalgae, oil shale, waste plastics, and polymer etc. The DAEM with a single distribution of activation energies cannot describe those reactions well since the thermal decomposition normally involves multiple sub-processes of various components. The double DAEM employs a double distribution to represent the activation energies. The Gaussian distribution is usually used to represent the activation energies. However, it is not sufficiently accurate for addressing the activation energies in the initial and final stages of the thermal decomposition reactions of solid fuels. Compared to the Gaussian distribution, the logistic distribution is slightly thicker at the curve tail and suits better to describe the activation energy distribution. In this work, a theoretical analysis of the double logistic DAEM for the thermal decomposition kinetics of solid fuels has been systematically investigated. After the derivation of the double logistic DAEM, its numerical calculation method and the physical meanings of the model parameters have been presented. Three typical types of simulated double logistic DAEM processes have been obtained according to the overlapped situation of two derivative conversion peaks, namely separated, overlapped and partially overlapped processes. It is found that, for the partially overlapped process, the form of the minor peak (overlapped peak or peak shoulder) depends on the values of the frequency factor and heating rate. Considering the simulated processes and related examples from literature, the double logistic DAEM has been remarked as a more reliable tool with abundant flexibility to explain the thermal decomposition of various solid fuels. More accurate results are expected if the double logistic DAEM is coupled with the computational fluid dynamics (CFD) simulation for those reactions mentioned above

PoSynDA: Multi-Hypothesis Pose Synthesis Domain Adaptation for Robust 3D Human Pose Estimation

Existing 3D human pose estimators face challenges in adapting to new datasets due to the lack of 2D-3D pose pairs in training sets. To overcome this issue, we propose \textit{Multi-Hypothesis \textbf{P}ose \textbf{Syn}thesis \textbf{D}omain \textbf{A}daptation} (\textbf{PoSynDA}) framework to bridge this data disparity gap in target domain. Typically, PoSynDA uses a diffusion-inspired structure to simulate 3D pose distribution in the target domain. By incorporating a multi-hypothesis network, PoSynDA generates diverse pose hypotheses and aligns them with the target domain. To do this, it first utilizes target-specific source augmentation to obtain the target domain distribution data from the source domain by decoupling the scale and position parameters. The process is then further refined through the teacher-student paradigm and low-rank adaptation. With extensive comparison of benchmarks such as Human3.6M and MPI-INF-3DHP, PoSynDA demonstrates competitive performance, even comparable to the target-trained MixSTE model\cite{zhang2022mixste}. This work paves the way for the practical application of 3D human pose estimation in unseen domains. The code is available at https://github.com/hbing-l/PoSynDA.Comment: Accepted to ACM Multimedia 2023; 10 pages, 4 figures, 8 tables; the code is at https://github.com/hbing-l/PoSynD

Molecular Modeling Study of Chiral Separation and Recognition Mechanism of β-Adrenergic Antagonists by Capillary Electrophoresis

Chiral separations of five β-adrenergic antagonists (propranolol, esmolol, atenolol, metoprolol, and bisoprolol) were studied by capillary electrophoresis using six cyclodextrins (CDs) as the chiral selectors. Carboxymethylated-β-cyclodextrin (CM-β-CD) exhibited a higher enantioselectivity power compared to the other tested CDs. The influences of the concentration of CM-β-CD, buffer pH, buffer concentration, temperature, and applied voltage were investigated. The good chiral separation of five β-adrenergic antagonists was achieved using 50 mM Tris buffer at pH 4.0 containing 8 mM CM-β-CD with an applied voltage of 24 kV at 20 °C. In order to understand possible chiral recognition mechanisms of these racemates with CM-β-CD, host-guest binding procedures of CM-β-CD and these racemates were studied using the molecular docking software Autodock. The binding free energy was calculated using the Autodock semi-empirical binding free energy function. The results showed that the phenyl or naphthyl ring inserted in the hydrophobic cavity of CM-β-CD and the side chain was found to point out of the cyclodextrin rim. Hydrogen bonding between CM-β-CD and these racemates played an important role in the process of enantionseparation and a model of the hydrogen bonding interaction positions was constructed. The difference in hydrogen bonding formed with the –OH next to the chiral center of the analytes may help to increase chiral discrimination and gave rise to a bigger separation factor. In addition, the longer side chain in the hydrophobic phenyl ring of the enantiomer was not beneficial for enantioseparation and the chiral selectivity factor was found to correspond to the difference in binding free energy

Identifying potential anti-COVID-19 pharmacological components of traditional Chinese medicine Lianhuaqingwen capsule based on human exposure and ACE2 biochromatography screening

药学院吴彩胜副教授联合海军军医大学柴逸峰教授团队在连花清瘟胶囊防治新冠肺炎的药理活性成分和机制研究方面取得新进展，这项研究基于HRMS和智能非靶向数据挖掘技术，全面分析了对多次给药后人血浆和尿液中的连花清瘟胶囊成分，合成了全新的ACE2生物色谱固定相，筛选出连花清瘟胶囊提取物和人尿液样品潜在的ACE2靶向成分。这项研究是连花清瘟胶囊的人体暴露信息的首次报道，为其在抗COVID-19的药理活性成分和作用机制研究提供了化学和药理学理论依据。本研究证明基于人体暴露的研究策略可用于高效的发掘中草药中的药效活性物质。【Abstract】Lianhuaqingwen (LHQW) capsule, a herb medicine product, has been clinically proved to be effective in coronavirus disease 2019 (COVID-19) pneumonia treatment. However, human exposure to LHQW components and their pharmacological effects remain largely unknown. Hence, this study aimed to determine human exposure to LHQW components and their anti-COVID-19 pharmacological activities. Analysis of LHQW component profiles in human plasma and urine after repeated therapeutic dosing was conducted using a combination of HRMS and an untargeted data-mining approach, leading to detection of 132 LHQW prototype and metabolite components, which were absorbed via the gastrointestinal tract and formed via biotransformation in human, respectively. Together with data from screening by comprehensive 2D angiotensin-converting enzyme 2 (ACE2) biochromatography, 8 components in LHQW that were exposed to human and had potential ACE2 targeting ability were identified for further pharmacodynamic evaluation. Results show that rhein, forsythoside A, forsythoside I, neochlorogenic acid and its isomers exhibited high inhibitory effect on ACE2. For the first time, this study provides chemical and biochemical evidence for exploring molecular mechanisms of therapeutic effects of LHQW capsule for the treatment of COVID-19 patients based on the components exposed to human. It also demonstrates the utility of the human exposure-based approach to identify pharmaceutically active components in Chinese herb medicines.The authors would like to thank Prof. Chuan Li in Shanghai Institute of Materia Medica, Chinese Academy of Sciences (Shanghai, China) to provide biological samples and technical guidance. This research was supported by Natural Science Foundation of China, China, (Grant Nos. 81773688, U1903119, 81973291, and 81973275); Zhejiang University Special Scientific Research Fund for COVID-19 Prevention and Control, China; “Phospherus” Project of Shanghai Science and Technology Committee, China, (Grant Nos. 19QA1411500); National Major Scientific and Technological Special Project for "Significant New Drugs Development", China, (Grant No. 2020ZX09201005)

Lack of Trehalose Accelerates H2O2-Induced Candida albicans Apoptosis through Regulating Ca2+ Signaling Pathway and Caspase Activity

Trehalose is a non-reducing disaccharide and can be accumulated in response to heat or oxidative stresses in Candida albicans. Here we showed that a C. albicans tps1Δ mutant, which is deficient in trehalose synthesis, exhibited increased apoptosis rate upon H2O2 treatment together with an increase of intracellular Ca2+ level and caspase activity. When the intracellular Ca2+ level was stimulated by adding CaCl2 or A23187, both the apoptosis rate and caspase activity were increased. In contrast, the presence of two calcium chelators, EGTA and BAPTA, could attenuate these effects. Moreover, we investigated the role of Ca2+ pathway in C. albicans apoptosis and found that both calcineurin and the calcineurin-dependent transcription factor, Crz1p, mutants showed decreased apoptosis and caspase activity upon H2O2 treatment compared to the wild-type cells. Expression of CaMCA1, the only gene found encoding a C. albicans metacaspase, in calcineurin-deleted or Crz1p-deleted cells restored the cell sensitivity to H2O2. Our results suggest that Ca2+ and its downstream calcineurin/Crz1p/CaMCA1 pathway are involved in H2O2 -induced C. albicans apoptosis. Inhibition of this pathway might be the mechanism for the protective role of trehalose in C. albicans

Methodology of drug screening and target identification for new necroptosis inhibitors

Apoptosis has been considered as the only form of regulated cell death for a long time. However, a novel form of programmed cell death called necroptosis was recently reported. The process of necroptosis is regulated and plays a critical role in the occurrence and development of multiple human diseases. Thus, the study on the molecular mechanism of necroptosis and its effective inhibitors has been an attractive field for researchers. Herein, we introduce the molecular mechanism of necroptosis and focus on the literature about necroptosis drug screening in recent years. In addition, the identification of the critical drug targets of the necroptosis is also discussed. Keywords: Necroptosis, Inhibitors, Drug screening, Target identification, Revie