Elucidation of active components and target mechanism in Jinqiancao granules for the treatment of prostatitis and benign prostatic hyperplasia.

ETHNOPHARMACOLOGICAL RELEVANCE: Prostatitis and benign prostatic hyperplasia (BPH) are inflammations of the prostate gland, which surrounds the urethra in males. Jinqiancao granules are a traditional Chinese medicine used to treat kidney stones and this medicine consists of four herbs: Desmodium styracifolium (Osbeck) Merr., Pyrrosia calvata (Baker) Ching, Plantago asiatica L. and stigma of Zea mays L. AIM OF THE STUDY: We hypothesized that Jinqiancao granules could be a potential therapy for prostatitis and BPH, and this work aimed to elucidate active compounds in Jinqiancao granules and their target mechanisms for the potential treatment of the two diseases. MATERIALS AND METHODS: Jinqiancao granules were commercially available and purchased. Database-driven data mining and networking were utilized to establish a general correlation between Jinqiancao granules and the two diseases above. Ultra-performance liquid chromatography-mass spectrometry was used for compound separation and characterization. The characterized compounds were evaluated on four G-protein coupled receptors (GPCRs: GPR35, muscarinic acetylcholine receptor M3, alpha-1A adrenergic receptor α1A and cannabinoid receptor CB2). A dynamic mass redistribution technique was applied to evaluate compounds on four GPCRs. Nitric acid (NO) inhibition was tested on the macrophage cell line RAW264.7. Molecular docking was conducted on GPR35-active compounds and GPR35 crystal structure. Statistical analysis using GEO datasets was conducted. RESULTS: Seventy compounds were isolated and twelve showed GPCR activity. Three compounds showed potent GPR35 agonistic activity (EC50 < 10 µM) and the GPR35 agonism action of PAL-21 (Scutellarein) was reported for the first time. Docking results revealed that the GPR35-targeting compounds interacted at the key residues for the agonist-initiated activation of GPR35. Five compounds showed weak antagonistic activity on M3, which was confirmed to be a disease target by statistical analysis. Seventeen compounds showed NO inhibitory activity. Several compounds showed multi-target properties. An experiment-based network reflected a pharmacological relationship between Jinqiancao granules and the two diseases. CONCLUSIONS: This study identified active compounds in Jinqiancao granules that have synergistic mechanisms, contributing to anti-inflammatory effects. The findings provide scientific evidence for the potential use of Jinqiancao granules as a treatment for prostatitis and BPH.

Screening for a Fenpropidin Enantiomer with High Activity and Low Toxicity.

Fenpropidin has been extensively used for managing fungal diseases in different crops. There is a lack of literature on the enantioselective bioactivity and toxicity of fenpropidin. This study aims to explore the enantioselective bioactivity and toxicity of fenpropidin. R-Fenpropidin exhibited more potent bioactivity against seven plant pathogens than S-fenpropidin. R-Fenpropidin was more effective than S-fenpropidin in inhibiting sclerotial production, affecting mycelial growth and morphology, increasing cell membrane permeability, and decreasing the ergosterol content of Rhizoctonia solani. R-Fenpropidin exhibited a tighter binding affinity and formed hydrogen bonds with two target proteins. Fenpropidin also has enantioselective toxicity to Selenastrum capricornutum, with the toxicity of S-fenpropidin being seven times that of R-fenpropidin. S-Fenpropidin significantly reduced the content of the photosynthetic pigments. The results showed that R-fenpropidin was a highly active enantiomer with low toxicity. This study can provide a basis for the development of enantiomers with high activity and low toxicity.

The dual-targeting mechanism of an anti-inflammatory diarylheptanoid from Curcuma zedoaria (Christm.) Roscoe with the capacity for ß2-adrenoreceptor agonism and NLRP3 inhibition.

Chronic obstructive pulmonary disease (COPD) is a common respiratory disease characterized by symptoms of shortness of breath and chronic inflammation. Curcuma zedoaria (Christm.) Roscoe is a well-documented traditional medical herb that is frequently used in the treatment of COPD. Previously, we identified a diarylheptanoid compound (1-(4-hydroxy-5-methoxyphenyl)-7-(4,5-dihydroxyphenyl)-3,5-dihydroxyheptane; abbreviated as HMDD) from this herb that exhibited potent agonistic activity on ß2-adrenergic receptors (ß2 adrenoreceptor) that are present on airway smooth muscle cells. In this work, we used chemically synthesized HMDD compound, and confirmed its bioactivity on ß2 adrenoreceptors. Then by a proteomics study and anti-inflammatory evaluation detections, we found that HMDD downregulated the nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) signaling pathway and suppressed the NLRP3 receptor expression in RAW264.7 macrophages and in a COPD model in A549 lung carcinoma cells. HMDD also decreased nitric oxide production levels, and impacted other interleukins and the phosphorylation of NF-κB and ERK pathways. We performed molecular docking of HMDD on ß2 adrenoreceptor and NLRP3 protein models. This work reports the anti-inflammatory effects of HMDD and suggests a dual-targeting mechanism of ß2-adrenoreceptor agonism and NLRP3 inhibition. Such a mechanism scientifically supports the clinical uses of Curcuma zedoaria (Christm.) Roscoe in treating COPD, as it can simultaneously relieve persistent breathlessness and inflammation. HMDD can be considered as a potential non-steroidal anti-inflammatory drug in novel therapy design for the treatment of COPD and other inflammatory diseases.

Specific biomarker mining and rapid detection of Burkholderia cepacia complex by recombinase polymerase amplification.

Objective: To mine specific proteins and their protein-coding genes as suitable molecular biomarkers for the Burkholderia cepacia Complex (BCC) bacteria detection based on mega analysis of microbial proteomic and genomic data comparisons and to develop a real-time recombinase polymerase amplification (rt-RPA) assay for rapid isothermal screening for pharmaceutical and personal care products. Methods: We constructed an automatic screening framework based on Python to compare the microbial proteomes of 78 BCC strains and 263 non-BCC strains to identify BCC-specific protein sequences. In addition, the specific protein-coding gene and its core DNA sequence were validated in silico with a self-built genome database containing 158 thousand bacteria. The appropriate methodology for BCC detection using rt-RPA was evaluated by 58 strains in pure culture and 33 batches of artificially contaminated pharmaceutical and personal care products. Results: We identified the protein SecY and its protein-coding gene secY through the automatic comparison framework. The virtual evaluation of the conserved region of the secY gene showed more than 99.8% specificity from the genome database, and it can distinguish all known BCC species from other bacteria by phylogenetic analysis. Furthermore, the detection limit of the rt-RPA assay targeting the secY gene was 5.6 × 102 CFU of BCC bacteria in pure culture or 1.2 pg of BCC bacteria genomic DNA within 30 min. It was validated to detect <1 CFU/portion of BCC bacteria from artificially contaminated samples after a pre-enrichment process. The relative trueness and sensitivity of the rt-RPA assay were 100% in practice compared to the reference methods. Conclusion: The automatic comparison framework for molecular biomarker mining is straightforward, universal, applicable, and efficient. Based on recognizing the BCC-specific protein SecY and its gene, we successfully established the rt-RPA assay for rapid detection in pharmaceutical and personal care products.

Protective effect of zerumbone on sepsis-induced acute lung injury through anti-inflammatory and antioxidative activity via NF-κB pathway inhibition and HO-1 activation.

Sepsis is a systemic illness for which there are no effective preventive or therapeutic therapies. Zerumbone, a natural molecule, has anti-oxidative and anti-inflammatory properties that may help to prevent sepsis. In the present study, we have assessed the protective effect of zerumbone against sepsis-induced acute lung injury (ALI) and its underlying mechanisms. During the experiment, mice were divided into five groups: a sham group, a sepsis-induced ALI group, and three sepsis groups that are pre-treated with zerumbone at different concentrations. We found that zerumbone greatly decreased the sepsis-induced ALI using histological investigations. Also, zerumbone treatment reduced the sepsis-induced inflammatory cytokine concentrations as well as the number of infiltrating inflammatory cells in BALF compared to non-treated sepsis animals. The zerumbone-pretreated sepsis groups had reduced pulmonary myeloperoxidase (MPO) activity than the sepsis groups. Moreover, the mechanism underlying the protective action of zerumbone on sepsis is accomplished by the activation of antioxidant genes such as nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), superoxide dismutase (SOD), and heme oxygenase 1 (HO-1). The obtained results revealed that zerumbone inhibited the sepsis-induced ALI through its anti-inflammatory and antioxidative activity via inhibition of the NF-κB pathway and activation of HO-1 pathway. Our findings demonstrate that zerumbone pretreatment suppresses sepsis-induced ALI via antioxidative activities and anti-inflammatory, implying that zerumbone could be a viable preventive agent for sepsis-induced ALI.

[Research progress of coarse-grained molecular dynamics in drug carrier materials].

As one of the traditional computer simulation techniques, molecular simulation can intuitively display and quantify molecular structure and explain experimental phenomena from the microscopic molecular level. When the simulation system increases, the amount of calculation will also increase, which will cause a great burden on the simulation system. Coarse-grained molecular dynamics is a method of mesoscopic molecular simulation, which can simplify the molecular structure and improve computational efficiency, as a result, coarse-grained molecular dynamics is often used when simulating macromolecular systems such as drug carrier materials. In this article, we reviewed the recent research results of using coarse-grained molecular dynamics to simulate drug carriers, in order to provide a reference for future pharmaceutical preparation research and accelerate the entry of drug research into the era of precision drug design.

Enantioselective uptake, translocation, and biotransformation of pydiflumetofen in wheat (Triticum aestivum L.): Insights from chiral profiling and molecular simulation.

Pydiflumetofen (PYD), a highly effective and broad-spectrum fungicide, is commonly employed for the control of fungal diseases. In this study, the uptake, translocation, and biotransformation of PYD by wheat (Triticum aestivum L.) were firstly investigated at a chiral level. The findings revealed that the residue concentration of R-PYD in wheat was higher than that of S-PYD, because of its higher uptake rate (k1 = 0.0421 h-1) and lower elimination rate (k2 = 0.0459 h-1). Additionally, R-PYD exhibited higher root bioconcentration factors and translocation factors compared with S-enantiomer, indicating R-PYD was more easily accumulating in roots and translocating to shoots. Furthermore, a total of 9 metabolites, including hydroxylated, demethylated, demethoxylated, dechlorinated, hydrolyzed, and glycosylated-conjugated products, were detected qualitatively in wheat roots or shoots. Symplastic pathway-mediated uptake, which predominantly relied on aquaporins and anion channels, was confirmed by root adsorption and inhibition experiments, without displaying any enantioselective effect. Molecular simulations demonstrated that R-PYD exhibited stronger binding affinity with TaLTP 1.1 with a lower grid score (-6.79 kcal/mol), whereas weaker interaction with the metabolic enzyme (CYP71C6v1) compared to the S-enantiomer. These findings highlight the significance of plant biomacromolecules in the enantioselective bioaccumulation and biotransformation processes. Importantly, a combination of experimental and theoretical evidence provide a comprehensive understanding of the fate of chiral pesticides in plants from an enantioselective perspective.

Fluoxetine Ameliorates the Aggravation of UC Symptoms in C57BL/6 Mice Induced by CUMS.

OBJECTIVE: Patients with chronic ulcerative colitis (UC) often have mental symptoms such as depression and anxiety, and stress can lead to gastrointestinal diseases. However, the correlation between mental stress and UC is unclear. In this paper, chronic unpredictable mild stress (CUMS) was utilized to evaluate the involvement of mental factors in the pathogenesis of UC. METHODS: The CUMS model was used to evaluate the direct/indirect involvement of mental factors in the pathogenesis of UC. The behavior was evaluated by the open field, forced swimming, and tail suspension tests. Body weight, the disease activity index (DAI) score, colon length, and HE staining of colon tissue were used to evaluate the action of CUMS and fluoxetine. RESULTS: The results showed that weight loss and the DAI score increased in CUMS mice, but they had no meaningful effect on colon length and morphological structure of colon tissue. However, CUMS aggravated dextran sulfate sodium (DSS)-induced colon length shortening and colon morphological structure damage. Fluoxetine significantly improved the DAI score, shortened colon length, and damaged morphology and structure of the colons induced by CUMS combined with DSS in mice. Fluoxetine also decreased the level of IL-6 in the serum and the TNF-α and IFN-γ levels of colon tissue. Fluoxetine simultaneously improved behavioral abnormalities induced by CUMS combined with DSS in mice. CONCLUSION: CUMS aggravated the UC symptoms induced by DSS, and fluoxetine could improve the UC symptoms due to its improvement in the inflammatory level and behavioral abnormalities.

Ciclopirox mitigates inflammatory response in LPS-induced septic shock via inactivation of SORT1-mediated wnt/ß-Catenin signaling pathway.

OBJECTIVE: Septic shock, the most severe stage of sepsis, is a deadly inflammatory disorder with high mortality. Ciclopirox (CPX) is a broad-spectrum antimycotic agent which also exerts anti-inflammatory effects in human diseases. However, whether CPX can relieve inflammatory response in LPS-induced septic shock remains unclear. MATERIALS AND METHODS: Male C57BL/6 mice LPS were injected intraperitoneally with LPS to simulate septic shock in vivo. RAW264.7 cells and bone marrow-derived macrophages (BMDMs) were subject to LPS treatment to simulate septic shock in vitro. ELISA was applied to detect the level of pro-inflammatory cytokines. Cell viability was assessed by CCK-8 assay. Protein levels was detected by western blotting. RESULTS: CPX enhanced the survival rate and attenuated inflammation in mice with LPS-induced septic shock. Similarly, CPX dose-dependently mitigated LPS-induced inflammation in BMDMs. It was also found that Sortilin 1 (SORT1) was upregulated in both in vivo and in vitro models of LPS-induced septic shock. In addition, SORT1 overexpression counteracted the alleviative effects of CPX on the inflammation response of LPS-challenged BMDMs by activating the Wnt/ß-Catenin signaling. Furthermore, BML-284 (a Wnt/ß-Catenin agonist) treatment also abrogated CPX-mediated moderation of LPS-triggered inflammatory reaction in BMDMs. CONCLUSIONS: In sum, we found that CPX protected against LPS-induced septic shock by mitigating inflammation via SORT1-mediated Wnt/ß-Catenin signaling pathway.

Deep-MSIM: Fast Image Reconstruction with Deep Learning in Multifocal Structured Illumination Microscopy.

Fast and precise reconstruction algorithm is desired for for multifocal structured illumination microscopy (MSIM) to obtain the super-resolution image. This work proposes a deep convolutional neural network (CNN) to learn a direct mapping from raw MSIM images to super-resolution image, which takes advantage of the computational advances of deep learning to accelerate the reconstruction. The method is validated on diverse biological structures and in vivo imaging of zebrafish at a depth of 100 µm. The results show that high-quality, super-resolution images can be reconstructed in one-third of the runtime consumed by conventional MSIM method, without compromising spatial resolution. Last but not least, a fourfold reduction in the number of raw images required for reconstruction is achieved by using the same network architecture, yet with different training data.

The enantioselective environmental fate of mandipropamid in water-sediment microcosms: Distribution, degradation, degradation pathways and toxicity assessment.

Mandipropamid (MDP) is a chiral fungicide that has been widely used to control oomycete pathogens. Currently, a comprehensive study of its environmental fate in aquatic ecosystems at the enantiomer level is still lacking. Here, the enantioselective environmental behaviors of MDP in four types of water-sediment microcosms were investigated. The concentrations of MDP enantiomers decreased over time in water due to sedimentation and degradation, while the concentrations in sediment increased to a peak and then slowly decreased because of adsorption and degradation. There were no enantioselective distribution behaviors in all kinds of microcosms. Moreover, R-MDP was preferentially degraded in lake water and Yangtze River water with half-lives of 5.92 d and 25.67 d, respectively. S-MDP was preferentially degraded in Yangtze River sediments, Yellow River sediments, and Yangtze River microcosm system, with half-lives ranging from 36.47 d to 77 d. Five degradation products of MDP in the sediment via hydrolysis and reduction were identified, and potential degradation pathways were proposed. Ecological Structure Activity Relationships (ECOSAR) prediction showed that the acute and chronic toxicities of all the products were higher than those of MDP except CGA 380778, which might pose a potential threat to aquatic ecosystems. This result offers new insights into the fate of chiral MDP in water-sediment ecosystems and will be conducive to the environmental and ecological risk assessment of MDP.

Microfluidic-Assisted Self-Assembly of 2D Nanosheets toward in situ Generation of Robust Nanofiber Film.

Methods allow the enhancement of nanofibers via self-assembly are potentially important for new disciplines with many advantages, including multi-anchor interaction, intrinsic mechanical properties and versatility. Herein, a microfluidic-assisted self-assembly process to construct hydroxyl functionalized boron nitride nanosheets (OH-BNNS)/graphene oxide (GO)/thermoplastic polyurethane (TPU) composite nanofiber film, in which stable and precisely controlled self-assembly is fulfilled by the confined ultra-small-volume chip is demonstrated. Multiple fine structural analyses alongside with the density-functional theory (DFT) calculations are implemented to confirm the synergistic effect of noncovalent interactions (hydrogen bonding interaction, π - π stacking interaction, and van der Waals attraction) plays a critical role in the robust micro-structure and a massive 700% enhancement of mechanical strength via adding only 0.3 wt% OH-BNNS and GO. Importantly, profiled from broadband optical absorption ability, robust mechanical properties and outstanding flexibility, the self-assembled 3D OH-BNNS/GO/TPU nanofiber film reveals an adorable evaporation rate of 4.04 kg m-2  h-1 under one sun illumination with stable energy transfer efficiency (93.2%) by accompanying hydrogen bonding interaction. This microfluidic-assisted self-assembly strategy will provide a constructive entry point for the rational design of nanofibers and beyond.

Comprehensive Assessment of Enantioselective Bioactivity, Toxicity, and Dissipation in Soil of the Chiral Herbicide Flurtamone.

Flurtamone is a typical chiral pesticide with a pair of enantiomers. In this study, the enantioselective biological effects of flurtamone enantiomers were systematically investigated. The bioactivities of R-flurtamone were 6.3-35.6 and 1.7-9.9 times higher than S- and Rac-flurtamone, respectively. The contribution of the R-enantiomer to herbicidal activity was 86.3-97.3%. The carotenoid content in the R-flurtamone treatment group was 1.31-2.14 times lower than that in the S-flurtamone treatment group. Molecular docking found that flurtamone can form aromatic H-bonds with phenylalanine (PHE) 233 of phytoene desaturase (PDS), while R-flurtamone can form π-π stacking with PHE 202 and PHE 311. The binding energies of R- and S-flurtamone were -10.239 and -7.555 kcal/mol, respectively. Furthermore, the acute toxicity levels to Selenastrum capricornutum of R- and Rac-flurtamone were both highly toxic, and S-flurtamone has moderate toxicity. S-Flurtamone dissipated preferentially. Therefore, the development of R-flurtamone can decrease environmental risks and protect human health.

Behavioral abnormalities in C57BL/6 mice with chronic ulcerative colitis induced by DSS.

BACKGROUND: Clinical epidemiological studies have found that some patients with ulcerative colitis (UC) are prone to mental disorders. DSS-induced acute and chronic UC models are often used to evaluate the efficacy of anti-UC drugs. However, whether DSS has an effect on mouse behavior has not been reported. METHODS: Acute and chronic UC models were induced by 3% DSS and 1.5% DSS, respectively. The bloody stool, the changes in the colon length, and histopathological changes in the colon were used to evaluate the success of the animal model. The behavior of mice was evaluated by open field experiment, tail suspension experiment and Sucrose preference test. RESULTS: The weight of mice in 3% DSS group decreased significantly, the DAI score increased significantly, the colon length of mice was significantly shortened, and the structure of colonic crypts was abnormal, which showed inflammatory cell infiltration and shrinkage of crypts. Compared with the control group, the immobility time of 3%DSS group mice in the tail suspension test and forced swimming test had no effect, the number of running and grooming times was significantly reduced, and there was no significant difference in the number of standing times. No abnormality was observed in HE staining of the hippocampus. However, in 1.5% DSS-induced chronic UC model, behavioral and hippocampal abnormalities were observed not only UC symptoms. CONCLUSIONS: The acute UC model induced by 3% DSS has certain influence on the behavior of mice, but the mental state of mice is normal, which may be the abnormal behavior caused by UC symptoms; However, the chronic UC model induced by 1.5% DSS has a significant effect on the behavior of mice, and the mice have mental disorders, which are caused by mental disorders.

Snapshot temporal compressive light-sheet fluorescence microscopy via deep denoising and total variation priors.

We present a snapshot temporal compressive light-sheet fluorescence microscopy system to capture high-speed microscopic scenes with a low-speed camera. A deep denoising network and total variation denoiser are incorporated into a plug-and-play framework to quickly reconstruct 20 high-speed video frames from a short-time measurement. Specifically, we can observe 1,000-frames-per-second (fps) microscopic scenes when the camera works at 50 fps to capture the measurement. The proposed method can potentially be applied to observe cell and tissue motions in thick living biological specimens.

Combined toxicity of acetamiprid and cadmium to larval zebrafish (Danio rerio) based on metabolomic analysis.

Emerging contaminants, such as neonicotinoid pesticide acetamiprid (Ace), are frequently detected in the water environment, which can interact with existing heavy metal cadmium (Cd) to produce unpredicted influence. Limited studies have evaluated the effects of multiple pollutant exposures on aquatic animals. Here, we characterized the joint toxicity of Ace and Cd exposure to zebrafish (Danio rerio). The results revealed that Cd and its combined exposure with Ace had an inhibitory effect on the growth of larval zebrafish and induced morphological defects. Combined exposure to high doses of Ace and Cd could significantly reduce the levels of TG, glucose, and pyruvate in larval zebrafish. Untargeted metabolomics revealed that Cd treatment (285) produced more differentially expressed metabolites (DEMs) than Ace treatment (115), and combined treatment produced the most DEMs (294). The KEGG pathway enrichment analysis showed that they could disrupt riboflavin metabolism, amino acid metabolism, and glycolipid metabolism in the larvae of D. rerio. ELISA showed that VB2, FMN, and FAD levels were significantly increased. In addition, gene expression analysis exhibited that the mRNA levels of essential genes related to glycolipid metabolism were substantially affected, such as PK, PEPckc, PPAR-α, and FABP6. Furthermore, targeted amino acid metabolomics confirmed that both single exposure to Cd and combined exposure to Ace and Cd altered the levels of amino acids in larvae, including ALA, ARG, MET, PRO, TYR, VAL, GLY, ORN, and PHE. Taken together, exposure to Ace and Cd, alone or in combination, exerted harmful effects on the individual development, riboflavin metabolism, glycolipid metabolism, and amino acid metabolism disorder of D. rerio. These findings highlighted that more attention should be paid to the compound toxicity of chemical mixtures to aquatic organisms.

Mucosal-Associated Invariant T Cells in the Digestive System: Defender or Destroyer?

Mucosal-associated invariant T (MAIT) cells are a subset of innate T lymphocytes that express the semi-invariant T cell receptor and recognize riboflavin metabolites via the major histocompatibility complex class I-related protein. Given the abundance of MAIT cells in the human body, their role in human diseases has been increasingly studied in recent years. MAIT cells may serve as targets for clinical therapy. Specifically, this review discusses how MAIT cells are altered in gastric, esophageal, intestinal, and hepatobiliary diseases and describes their protective or pathogenic roles. A greater understanding of MAIT cells will provide a more favorable therapeutic approach for digestive diseases in the clinical field.

Revealing the structure and organization of intercellular tunneling nanotubes (TNTs) by STORM imaging.

Tunneling nanotubes (TNTs) are nanoscale, actin-rich, transient intercellular tubes for cell-to-cell communication, which transport various cargoes between distant cells. The structural complexity and spatial organization of the involved components of TNTs remain unknown. In this work, the STORM super-resolution imaging technique was applied to elucidate the structural organization of microfilaments and microtubules in intercellular TNTs at the nanometer scale. Our results reveal different distributions of microfilaments and intertwined structures of microtubules in TNTs, which promote the knowledge of TNT communications.

Integration of Two-Dimensional Liquid Chromatography-Mass Spectrometry and Molecular Docking to Characterize and Predict Polar Active Compounds in Curcuma kwangsiensis.

Curcuma kwangsiensis, one species of Curcumae zedoaria Ros. c, is a commonly used traditional Chinese medicine (TCM) for treating cardiovascular disease, cancer, asthma and inflammation. Polar compounds are abundant in water decoction, which would be responsible for critical pharmacological effects. However, current research on polar compounds in Curcumae zedoaria Ros. c remains scarce. In this study, the polar fraction from Curcuma kwangsiensis was firstly profiled on G protein-coupled receptor 109A (GPR109A), ß2-adrenergic receptor (ß2-AR), neurotensin receptor (NTSR), muscarinic-3 acetylcholine receptor (M3) and G protein-coupled receptor 35 (GPR35), which were involved in its clinical indications and exhibited excellent ß2-AR and GPR109A receptor activities. Then, an offline two-dimensional reversed-phase liquid chromatography (RPLC) coupled with the hydrophilic interaction chromatography (HILIC) method was developed to separate polar compounds. By the combination of a polar-copolymerized XAqua C18 column and an amide-bonded XAmide column, an orthogonality of 47.6% was achieved. As a result of coupling with the mass spectrometry (MS), a four-dimensional data plot was presented in which 373 mass peaks were detected and 22 polar compounds tentatively identified, including the GPR109A agonist niacin. Finally, molecular docking of these 22 identified compounds to ß2-AR, M3, GPR35 and GPR109A receptors was performed to predict potential active ingredients, and compound 9 was predicted to have a similar interaction to the ß2-AR partial agonist salmeterol. These results were supplementary to the material basis of Curcuma kwangsiensis and facilitated the bioactivity research of polar compounds. The integration of RPLC×HILIC-MS and molecular docking can be a powerful tool for characterizing and predicting polar active components in TCM.

Stereoselective toxicity, bioaccumulation, and metabolic pathways of triazole fungicide cyproconazole in zebrafish.

Cyproconazole (CPZ) is a broad-spectrum fungicide that is widely used around the world. CPZ can persist in water which raised concerns about its potential adverse effects on aquatic life. In this study, the stereoselective toxicity, bioaccumulation, elimination, and kinetic biotransformation in zebrafish were investigated. The LC50 of 96 h acute toxicity was 15.88, 19.68, 26.99, and 17.10 mg/L for SR-, SS-, RS-, and RR-CPZ, respectively. The uptake and elimination experiment showed the bioconcentration factor in order of SR- > RR- > SS- > RS-CPZ at the exposure concentration of 0.1 and 1 mg/L. In the depuration stage, CPZ isomers were rapidly eliminated by 99% within 24 h. Moreover, the oxidative stress responses (POD, SOD, and CAT) were stereoselectively induced by CPZ stereoisomers, the activity of POD was significantly increased in all CPZ treatment groups compared to the control while the activity of CAT exhibited a concentration-dependent decrease in the CPZ treatment group. Multiple metabolic pathways of CPZ in zebrafish were proposed for the first time and 7 phase I metabolites and 25 phase II conjugates were found. This study determined the potential toxicity of CPZ to zebrafish and provided a strategy for the risk evaluation of CPZ stereoisomers in aquatic ecosystems.