Exploring antithrombotic mechanisms and effective constituents of Lagopsis supina using an integrated strategy based on network pharmacology, molecular docking, metabolomics, and experimental verification in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Thrombosis is a common cause of morbidity and mortality worldwide. Lagopsis supina (Stephan ex Willd.) Ikonn.-Gal. ex Knorring is an ancient Chinese herbal medicine used for treating thrombotic diseases. Nevertheless, the antithrombotic mechanisms and effective constituents of this plant have not been clarified. AIM OF THE STUDY: This work aimed to elucidate the pharmacodynamics and mechanism of L. supina against thrombosis. MATERIALS AND METHODS: Systematic network pharmacology was used to explore candidate effective constituents and hub targets of L. supina against thrombosis. Subsequently, the binding affinities of major constituents with core targets were verified by molecular docking analysis. Afterward, the therapeutic effect and mechanism were evaluated in an arteriovenous bypass thrombosis rat model. In addition, the serum metabolomics analysis was conducted using ultra-high performance liquid chromatography coupled with Q-Exactive mass spectrometry. RESULTS: A total of 124 intersected targets of L. supina against thrombosis were predicted. Among them, 24 hub targets were obtained and their mainly associated with inflammation, angiogenesis, and thrombosis approaches. Furthermore, 9 candidate effective constituents, including (22E,24R)-5α,8α-epidioxyergosta-6,22-dien-3ß-ol, aurantiamide, (22E,24R)-5α,8α-epidioxyergosta-6,9 (11),22-trien-3ß-ol, lagopsinA, lagopsin C, 15-epi-lagopsin C, lagopsin D, 15-epi-lagopsin D, and lagopsin G in L. supina and 6 potential core targets (TLR-4, TNF-α, HIF-1α, VEGF-A, VEGFR-2, and CLEC1B) were acquired. Then, these 9 constituents demonstrated strong binding affinities with the 6 targets, with their lowest binding energies were all less than -5.0 kcal/mol. The antithrombotic effect and potential mechanisms of L. supina were verified, showing a positively associated with the inhibition of inflammation (TNF-α, IL-1ß, IL-6, IL-8, and IL-10) and coagulation cascade (TT, APTT, PT, FIB, AT-III), promotion of angiogenesis (VEGF), suppression of platelet activation (TXB2, 6-keto-PGF1α, and TXB2/6-keto-PGF1α), and prevention of fibrinolysis (t-PA, u-PA, PAI-1, PAI-1/t-PA, PAI-1/u-PA, and PLG). Finally, 14 endogenous differential metabolites from serum samples of rats were intervened by L. supina based on untargeted metabolomics analysis, which were closely related to amino acid metabolism, inflammatory and angiogenic pathways. CONCLUSION: Our integrated strategy based on network pharmacology, molecular docking, metabolomics, and in vivo experiments revealed for the first time that L. supina exerts a significant antithrombotic effect through the inhibition of inflammation and coagulation cascade, promotion of angiogenesis, and suppression of platelet activation. This paper provides novel insight into the potential of L. supina as a candidate agent to treat thrombosis.

Association between Visceral Adiposity Index and Hyperuricemia among Steelworkers: The Moderating Effects of Drinking Tea.

BACKGROUND/OBJECTIVES: Steelworkers are more likely to have a higher prevalence of hyperuricemia due to their exposure to special occupational factors and dietary habits. The interrelationships of visceral adiposity index (VAI), hyperuricemia, and drinking tea remain uncertain. This study aimed to assess the association between VAI and hyperuricemia among steelworkers, and if drinking tea modified this association. METHODS: A total of 9928 steelworkers from Hunan Hualing Xiangtan Iron and Steel Company participated in this cross-sectional study. All participants completed a questionnaire, received anthropometric measurements, and provided blood samples for biochemical testing. Three logistic regression models were used to analyze the association between VAI and hyperuricemia. RESULTS: In this study, the prevalence of hyperuricemia was approximately 23.74% (males: 24.41%; females: 20.63%), and a positive correlation between VAI and hyperuricemia risk was observed. In multivariate logistic regression analysis, the risk of hyperuricemia increased 1.76 times (95% CI: 1.64-1.89) and 2.13 times (95% CI: 1.76-2.57) with the increase of ln VAI in males and females, respectively. For males, compared to quartile 1, the risk of hyperuricemia in the second, third, and fourth quartile of VAI were 1.75 (95% CI: 1.11-2.71), 2.56 (95% CI: 1.67-3.93) and 4.89 (95% CI: 3.22-7.43). For females, compared to quartile 1, the risk of hyperuricemia in the second, third, and fourth quartile of VAI were 1.99 (95% CI: 1.40-2.82), 2.92 (95% CI: 1.96-4.34) and 4.51 (95% CI: 2.89-7.02). Additionally, our study found that, compared with not consuming tea, drinking tea could reduce uric acid levels by 0.014 in male steelworkers (t = -2.051, p = 0.040), 0.020 in workers consuming smoked food (t = -2.569, p = 0.010), and 0.022 in workers consuming pickled food (t = -2.764, p = 0.006). CONCLUSIONS: In conclusion, VAI is positively correlated with hyperuricemia in steelworkers. Drinking tea may lower uric acid levels in male steelworkers and steelworkers who prefer smoked and pickled foods.

Insight into the mechanism of alkali-thermal pretreatment of food-waste solid residue through fluorescence spectroscopy coupled with parallel factor analysis.

Food-waste solid residue is the remaining solid after food waste treatment, with high yield, high solid content, high protein and fiber content. Effective pretreatment is necessary to improve the efficiency of hydrolysis and acidification for anaerobic digestion of food-waste solid residue. In this study, fluorescence spectroscopy coupled with parallel factor analysis were used to insight into the mechanism of food-waste solid residue during three pretreatments (alkali, thermal and alkali-thermal). Pretreatments increased the solubility of lignocellulosic substrate and destroyed structure of starch, while lignocellulosic analogs were effectively cracked, changing the composition and improving the degradability. Soluble chemical oxygen demand, soluble protein and soluble polysaccharide concentrations were increased by 144.60%, 350.57% and 138.72% after pretreatment under the condition of 120 °C + 2% CaO, respectively. Three-dimensional fluorescence spectra showed the region of maximum fluorescence intensity under alkali-thermal pretreatments, indicating chemical bonds (such as OC-C) were easier broken and the solubility of organic substances were increased. Three main fluorescence components were obtained by parallel factor analysis, which were humic acid-like, lignocellulose-like and protein-like, respectively, while the lignocellulose-like had the maximum Fmax value. The fluorescence intensity of samples under alkali-thermal pretreatment varied in the range from 59.48 × 105 to 13.18 × 106, which was an increase of 174.27%-507.74% over the control (21.68 × 105), indicating that alkali-thermal pretreatment observably accelerated the breaking of chemical bonds, and thus promoted the dissolution of organic matter. This study deeply revealed the mechanism of alkali-thermal pretreatment of food-waste solid residue, which is of great significance for efficient resource utilization of food waste and food-waste solid residue.

Enhanced adsorption of phosphate by rice straw-based biochar prepared via metal impregnation and bio-template technology.

Phosphate removal from water through green, highly efficient technologies has received much attention. Biochar is an effective adsorbent for phosphate removal. However, adsorption capacity of phosphate on pristine rice straw-based biochar was not optimistic due to low anion exchange capacity. In this study, Fe-modified, Mg-modified and MgFe-modified rice straw-based biochar (Fe-BC, Mg-BC and MgFe-BC) were prepared by combining metal impregnation and biological template methods to improve the adsorption capacity of phosphate. The surface characteristics of biochar and the adsorption behavior of phosphate on biochar were investigated. The modified biochar had the specific surface area of 17.910-39.336 m2/g, and their surfaces were rich in a large number of functional groups and metal oxides. Phosphate release was observed on pristine rice straw-based biochar without metal impregnation. The maximum adsorption capacities of phosphate on MgFe-BC, Mg-BC and Fe-BC at 298 K were 6.93, 5.75 and 0.23 mg/g, respectively. Adsorption was a spontaneous endothermic process, while chemical adsorption dominated and electrostatic attraction and pores filling existed simultaneously. Based on the site energy distribution theory study, the standard deviation of MgFe-BC decreased from 6.96 to 4.64 kJ/mol with temperature increasing, which proved that the higher the temperature would cause the lower heterogeneity. Moreover, the effects of pH, humic acid, co-existing ions and ionic strength on phosphate adsorption of MgFe-BC were also discussed. MgFe-BC with fine pores and efficient adsorption sites is an ideal adsorbent for phosphate removal from water.

Gene-gene interaction network analysis indicates CNTN2 is a candidate gene for idiopathic generalized epilepsy.

Twin and family studies have established the genetic contribution to idiopathic generalized epilepsy (IGE). The genetic architecture of IGE is generally complex and heterogeneous, and the majority of the genetic burden in IGE remains unsolved. We hypothesize that gene-gene interactions contribute to the complex inheritance of IGE. CNTN2 (OMIM* 615,400) variants have been identified in cases with familial adult myoclonic epilepsy and other epilepsies. To explore the gene-gene interaction network in IGE, we took the CNTN2 gene as an example and investigated its co-occurrent genetic variants in IGE cases. We performed whole-exome sequencing in 114 unrelated IGE cases and 296 healthy controls. Variants were qualified with sequencing quality, minor allele frequency, in silico prediction, genetic phenotype, and recurrent case numbers. The STRING_TOP25 gene interaction network analysis was introduced with the bait gene CNTN2 (denoted as A). The gene-gene interaction pair mode was presumed to be A + c, A + d, A + e, with a leading gene A, or A + B + f, A + B + g, A + B + h, with a double-gene A + B, or other combinations. We compared the number of gene interaction pairs between the case and control groups. We identified three pairs in the case group, CNTN2 + PTPN18, CNTN2 + CNTN1 + ANK2 + ANK3 + SNTG2, and CNTN2 + PTPRZ1, while we did not discover any pairs in the control group. The number of gene interaction pairs in the case group was much more than in the control group (p = 0.021). Taking together the genetic bioinformatics, reported epilepsy cases, and statistical evidence in the study, we supposed CNTN2 as a candidate pathogenic gene for IGE. The gene interaction network analysis might help screen candidate genes for IGE or other complex genetic disorders.

High performance polyvinyl alcohol/lignin fibers with excellent mechanical and water resistance properties.

To address the shortcoming of Polyvinyl alcohol (PVA) fibers for food or medical packaging materials including low mechanical strength and poor water resistance, lignin (LN) was used as raw material, acetone/H2O as solvent to self-assemble into lignin nanoparticles (LNP) by adverse solvent precipitation approach, and then PVA/LNP composite fibers with different LNP contents were fabricated successfully by wet and dry spinning. Herein, vast hydrophilic hydroxyl groups in PVA decreased owing to the hydrogen bond between LN and PVA, Especially, with only 0.5 wt% loading of LNP into the PVA/LNP fibers, the diameter was 94.4 dtex, tensile strength was 10.1 cN/dtex (1279.8 MPa), initial modulus was 94.7 cN/dtex (12.0 GPa), the crystallinity was 56.7 %, the orientation was 97.1 %, and water contact angle was 103.1°. Compared with pure PVA fibers, the tensile strength of PVA/LNP-0.5 fibers was increased by 44.2 % and the contact angle was increased 37°. This work provides novel insights into obtaining lignin-reinforced PVA composite fibers with strong mechanical properties and excellent water resistance properties, indicating the potential of the PVA/LNP fibers for food or medical packaging application.

A potential dual protection vaccine: Recombinant feline herpesvirus-1 expressing feline parvovirus VP2 antigen.

Recently, herpesvirus viral vectors that stimulate strong humoral and cellular immunity have been demonstrated to be the most promising platforms for the development of multivalent vaccines, because they contain various nonessential genes and exhibit long-life latency characteristics. Previously, we showed that the feline herpesvirus-1 (FHV-1) mutant WH2020-ΔTK/gI/gE, which was safe for felines and provided efficacious protection against FHV-1 challenge, can be used as a vaccine vector. Moreover, previous studies have shown that the major neutralizing epitope VP2 protein of feline parvovirus (FPV) can elicit high levels of neutralizing antibodies. Therefore, to develop a bivalent vaccine against FPV and FHV-1, we first generated a novel recombinant virus by CRISPR/Cas9-mediated homologous recombination, WH2020-ΔTK/gI/gE-VP2, which expresses the VP2 protein of FPV. The growth characteristics of WH2020-ΔTK/gI/gE-VP2 were similar to those of WH2020-ΔTK/gI/gE, and WH2020-ΔTK/gI/gE-VP2 was stable for at least 30 generations in CRFK cells. As expected, we found that the felines immunized with WH2020-ΔTK/gI/gE-VP2 produced FPV-neutralizing antibody titers (27.5) above the positive cutoff (26) on day 14 after single inoculation. More importantly, recombinant WH2020-ΔTK/gI/gE-VP2 exhibited severely impaired pathogenicity in inoculated and cohabiting cats. The kittens immunized with WH2020-ΔTK/gI/gE and WH2020-ΔTK/gI/gE-VP2 produced similar levels of FHV-specific antibodies and IFN-ß. Furthermore, felines immunized with WH2020-ΔTK/gI/gE-VP2 were protected against challenge with FPV and FHV-1. These data showed that WH2020-ΔTK/gI/gE-VP2 appears to be a potentially safe, effective, and economical bivalent vaccine against FPV and FHV-1 and that WH2020-ΔTK/gI/gE can be used as a viral vector to develop feline multivalent vaccines.

Chemical constituents isolated from Hosta ensata and their anti-inflammatory activities.

A new phenol derivative, hostaphenol A (1), along with 16 known ones (2-17) were isolated from an ethanolic extract of the whole plants of Hosta ensata F. Maek. Their structures were elucidated by HRMS and NMR data as well as comparison with those reported in literature. The report of the first cyclopeptide and compounds 5, 6, 8, 10, 12-15, and 17 in the Asparagaceae family. Compound 2, as well as compounds 3, 4, 7, 9, 11, and 16 were reported for the first time from the Hosta genus and this plant, respectively. All compounds significantly reduced nitric oxide (NO) production at a concentration of 40 µM with no toxicity in RAW 264.7 cells stimulated by lipopolysaccharide. Among them, compounds 2-5 (40 µM) exerted obvious NO inhibitory activities, and their inhibition rate was exceeded 50%.

Lagopsis supina ameliorates myocardial ischemia injury by regulating angiogenesis, thrombosis, inflammation, and energy metabolism through VEGF, ROS and HMGB1 signaling pathways in rats.

BACKGROUND: Lagopsis supina (Steph. ex. Willd.) Ikonn.-Gal. is an important traditional Chinese medicine used to treat various ailments. However, its impact on myocardial ischemia (MI) injury remains unknown. PURPOSE: This research aimed to reveal the therapeutic effect, potential mechanism, and metabolomics of L. supina against MI injury in rats. METHODS: The therapeutic effects of the ethanolic extract of L. supina (LS) and its four fractions (LSA∼D) on a left anterior descending (LAD) artery occlusion-induced MI model rat were explored. The pharmacodynamics including myocardial infraction area, myocardial tissue pathology and apoptosis, and serum biochemical parameters (CK, CK-MB, CTn-T, SOD, ET-1, NO, eNOS, VEGF, TXB2, 6-keto-PGF1α, TNF-α, IL-6, and CRP) were evaluated. The 24 related protein expressions were detected using western blotting assay. Simultaneously, the qualitative and quantitative analyses of microporous adsorption resin with 30% (LSC) and 60% (LSD) aqueous ethanol fractions were performed using UHPLC-MS and HPLC. Moreover, the serum metabolomics analysis of rats was profiled using UHPLC-MS. RESULTS: LS exerted remarkable alleviating effect on MI in rats. Importantly, LSC and LSD, two effective fractions of LS, significantly reduced myocardial infraction area, alleviated myocardial tissue pathology and apoptosis, regulated serum biochemical parameters. Furthermore, LSC and LSD markedly up-regulated the levels of VEGF-A, VEGFR-2, PKC, Bcl-2, Nrf2, HO-1, and thrombin, as well as prominently down-regulated the protein expression of Notch 1, p-PI3K, p-PI3K/PI3K, p-Akt, p-Akt/Akt, Bax, cleaved-caspase-3, cleaved-caspase-3/caspase-3, vWF, p-Erk, p-Erk/Erk, HMGB1, p-p38, p-p38/p38, p-p65, and p-p65/p65. A total of 26 candidate biomarkers were significantly regulated by LSC and LSD and they are mainly involved in amino acid metabolism, glycerophospholipid metabolism, and sphingolipid metabolism. Finally, phenylethanols and flavonoids may be major bio-constituents of LSC and LSD against MI. CONCLUSIONS: This work, for the first time, demonstrated that L. supina had a significant therapeutic effect on MI in rats. Additionally, LSC and LSD, two bio-fractions from L. supina, exerted their potential to ameliorate MI injury by promoting angiogenesis, inhibiting thrombosis, blocking inflammation, and facilitating energy metabolism through promotion of VEGF pathway, as well as suppression of ROS and HMGB1 pathways in rats. These findings suggest that LSC and LSD hold promise as potential therapeutic agents for MI injury in clinical application.

Rapid Identification of Alkaloids and Flavonoids in Fissistigma oldhamii var. longistipitatum by Ultra High-Performance Liquid Chromatography and Quadrupole Time-of-Flight Tandem Mass Spectrometry.

The herb Fissistigma oldhamii var. longistipitatum has been used for a long time in Asian folk medicine in the treatment of several diseases, including rheumatoid arthritis and other inflammatory conditions. Researchers in China and elsewhere have analyzed and characterized its chemical content. In this study, a UHPLC-Q-TOF-MS/MS method, run in both positive and negative modes, was used to identify the main chemical compounds in dichloromethane extracts of this F. oldhamii variant. A total of 64 compounds, including 44 alkaloids and 20 flavonoids, were rapidly identified or tentatively characterized by comparing the molecular ion peaks and MS2 mass spectrometry fragment ions, combined with the mass spectrometry information of reference substances, appropriate fragmentation ions and related literatures. For the first time, the developed UHPLC-Q-TOF-MS/MS analysis method allows for the determination of 64 compounds from extracts of the F. oldhamii variant. The method presented here produced results that will be useful in further studies of this herb.

Dirac cones in bipartite square-octagon lattice: A theoretical approach.

Dirac cones are difficult to achieve in a square lattice with full symmetry. Here, we have theoretically investigated a bipartite tetragonal lattice composed of tetragons and octagons using both Tight-Binding (TB) model and density functional theory (DFT) calculations. The TB model predicts that the system exhibits nodal line semi-metallic properties when the on-site energies of all atoms are identical. When the on-site energies differ, the formation of an elliptical Dirac cone is predicted. Its physical properties (anisotropy, tilting, merging, and emerging) can be regulated by the hopping energies. An exact analytical formula is derived to determine the position of the Dirac point by the TB parameters, and a criterion for the existence of Dirac cones is obtained. The "divide-and-coupling" method is applied to understand the origin of the Dirac cone, which involves dividing the bands into several groups and examining the couplings among inter-groups and intra-groups. Various practical systems computed by DFT methods, e.g., t-BN, t-Si, 4,12,2-graphyne, and t-SiC, are also examined, and they all possess nodal lines or Dirac cones as predicted by the TB model. The results provide theoretical foundation for designing novel Dirac materials with tetragonal symmetry.

Effects of incineration and pyrolysis on removal of organics and liberation of cathode active materials derived from spent ternary lithium-ion batteries.

Removing organics via thermal treatment to liberate active materials from spent cathode sheets is essential for recovering lithium-ion batteries. In this study, the effects of incineration, N2 pyrolysis, and CO2 pyrolysis on the removal of organics and liberation of ternary cathode active materials (CAMs) were compared. The results indicated that the organics in the spent ternary cathode sheets comprised a residual electrolyte and polyvinylidene fluoride (PVDF) binder. Moreover, the organics could be removed to promote the liberation of CAMs via incineration, N2 pyrolysis, and CO2 pyrolysis. When the temperature was <200 °C, the chemical properties of the volatilized ester electrolyte remained unchanged during both N2 and CO2 pyrolysis, indicating that the electrolyte can be collected by controlling the pyrolysis temperature and condensation. Furthermore, PVDF binder decomposition occurred at 200-600 °C. The optimal temperatures of incineration, N2 pyrolysis, and CO2 pyrolysis were 550, 500, and 450 °C, respectively, and these treatments increased the liberation efficiency of CAMs from 81.49 % to 98.75 %, 99.26 %, and 97.98 %, respectively. In addition, heat-treated CAMs required less time to achieve adequate liberation. Following three thermal treatment processes, the sizes of the CAM particles were mainly concentrated in the ranges of 0.075-0.1 mm and <0.075 mm. Furthermore, for all types of CAMs examined, the Al concentration decreased from 1.09 % to <0.35 %, which increased the separation efficiency and improved the chemical metallurgical performance.

Identification of N-(3-(methyl(3-(orotic amido)propyl)amino)propyl) oleanolamide as a novel topoisomerase I catalytic inhibitor by rational design, molecular dynamics simulation, and biological evaluation.

DNA topoisomerase I (TOP1) catalytic inhibitors are a promising class of antitumor agents. Oleanolic acid derivatives are potential TOP1 catalytic inhibitors. However, their inhibitory activity still needs to be enhanced, and the stability and hotspot residue sites of their interaction with TOP1 remain to be elucidated. Herein, a novel oleanolic acid derivative, OA4 (N-(3-(methyl(3-(orotic amido)propyl)amino)propyl)oleanolamide), was identified by rational design. Subsequently, molecular dynamics simulations were performed to explore the stability and conformational dynamics of the TOP1-OA4 complex. The molecular mechanics/generalized Born surface area method calculated the binding free energy and predicted Arg488, Ile535, and His632 to be hotspot residues. Biological experiments verified that OA4 is a nonintercalative TOP1 catalytic inhibitor. OA4 exhibits better proliferation inhibitory activity against tumor cells than normal cells. Furthermore, OA4 can induce apoptosis and effectively suppress the proliferation and migration of cancer cells. This work provides new insights for the development of novel TOP1 catalytic inhibitors.

The activities and mechanisms of intestinal microbiota metabolites of TCM herbal ingredients could be illustrated by a strategy integrating spectrum-effects, network pharmacology, metabolomics and molecular docking analysis: Platycodin D as an example.

BACKGROUND: The intestinal microbiota plays a key role in understanding the mechanism of traditional Chinese medicine (TCM), as it could transform the herbal ingredients to metabolites with higher bioavailability and activity comparing to their prototypes. Nevertheless, the study of the activity and mechanism of microbiota metabolites reported by the published literature still lacks viable ways. Hence a new strategy is proposed to solve this issue. PURPOSE: A new strategy to study the activity and mechanism of intestinal microbiota metabolites of TCM herbal ingredients by integrating spectrum-effect relationship, network pharmacology, metabolomics analysis and molecular docking together was developed and proposed. METHOD: Platycodin D (PD) and its microbiota metabolites with antitussive and expectorant effect were selected as an example for demonstration. First, the PD and its microbiota metabolites with important contribution to antitussive and/or expectorant effects were screened through spectrum-effect relationship analysis. Second, network pharmacology and metabolomics analysis were integrated to identify the upstream key targets of PD and its microbiota metabolites as well as the downstream endogenous metabolites. Finally, the active forms of PD were further confirmed by molecular docking. RESULTS: Results showed that PD was an active ingredient with antitussive and/or expectorant effects, and the active forms of PD were its microbiota metabolites: 3-O-ß-d-glucopyranosyl platycodigenin, 3-O-ß-d-glucopyranosyl isoplatycodigenin, 7­hydroxyl-3-O-ß-d-glucopyranosyl platycodigenin, platycodigenin and isoplatycodigenin. In addition, those microbiota metabolites could bind the key targets of PAH, PLA2G2A, ALOX5, CYP2C9 and CYP2D6 to exert antitussive effects by regulating four metabolic pathways of phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism, glycerophospholipid metabolism and linoleic acid metabolism. Similarly, they could also bind the key targets of PLA2G1B, ALOX5, CYP2C9 and CYP2D6 to exert expectorant effect by regulating two pathways of glycerophospholipid metabolism and linoleic acid metabolism. CONCLUSION: The proposed strategy paves a new way for the illustration of the activities and mechanisms of TCM herbal ingredients, which is very important to reconcile the conundrums of TCM herbal ingredients with low oral bioavailability but high activity.

Sub-region analysis of DMD gene in cases with idiopathic generalized epilepsy.

Gene sub-region encoded protein domain is the basic unit for protein structure and function. The DMD gene is the largest coding gene in humans, with its phenotype relevant to idiopathic generalized epilepsy. We hypothesized variants clustered in sub-regions of idiopathic generalized epilepsy genes and investigated the relationship between the DMD gene and idiopathic generalized epilepsy. Whole exome sequencing was performed in 106 idiopathic generalized epilepsy individuals. DMD variants were filtered with variant type, allele frequency, in silico prediction, hemizygous or homozygous status in the population, inheritance mode, and domain location. Variants located at the sub-regions were selected by the subRVIS software. The pathogenicity of variants was evaluated by the American College of Medical Genetics and Genomics criteria. Articles on functional studies related to epilepsy for variants clustered protein domains were reviewed. In sub-regions of the DMD gene, two variants were identified in two unrelated cases with juvenile absence epilepsy or juvenile myoclonic epilepsy. The pathogenicity of both variants was uncertain significance. Allele frequency of both variants in probands with idiopathic generalized epilepsy reached statistical significance compared with the population (Fisher's test, p = 2.02 × 10-6, adjusted α = 4.52 × 10-6). The variants clustered in the spectrin domain of dystrophin, which binds to glycoprotein complexes and indirectly affects ion channels contributing to epileptogenesis. Gene sub-region analysis suggests a weak association between the DMD gene and idiopathic generalized epilepsy. Functional analysis of gene sub-region helps infer the pathogenesis of idiopathic generalized epilepsy.

Anti-inflammatory constituents isolated from the flowers of Hosta plantaginea via suppression of the NF-κB signaling pathway in LPS-stimulated RAW 264.7 macrophages.

Hosta plantaginea (Lam.) Aschers flower is traditionally used in China as an important herbal medicine for the treatment of inflammatory disease. The present study isolated one new compound, namely (3R)-dihydrobonducellin (1), and five known ones, p-hydroxycinnamic acid (2), paprazine (3), thymidine (4), bis(2-ethylhexyl) phthalate (5), and dibutyl phthalate (6) from H. plantaginea flowers. These structures were elucidated from spectroscopic data. Among them, compounds 1-4 remarkably suppressed nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW 264.7 cells with half maximal inhibitory concentration (IC50) values of 19.88 ± 1.81, 39.80 ± 0.85, 19.03 ± 2.35, and 34.63 ± 2.38 µM, respectively. Furthermore, compounds 1 and 3 (20 µM) significantly decreased levels of tumor necrosis factor α (TNF-α), prostaglandin E2 (PGE2), interleukin 1ß (IL-1ß), and IL-6. Additionally, compounds 1 and 3 (20 µM) prominently reduced the phosphorylation protein level of nuclear factor kappa-B (NF-κB) p65. The present findings indicated that compounds 1 and 3 may be new candidates against inflammation via blocking the NF-κB signaling pathway.

Activating Blood Circulation, Anti-Inflammatory and Diuretic Effects of Leonurus japonicus Extract on a Rat Model of Trauma Blood Stasis and Its Phytochemical Profiling.

Leonurus japonicus Houtt. has been traditionally used to treat many ailments. This study evaluated the activating blood circulation, anti-inflammatory, and diuretic effects of L. japonicus extract (LJ) and identified its phytochemicals. In this work, the phytochemicals in LJ were identified using liquid chromatography mass spectrometry. Rats were randomly assigned to three groups (n=8): Control group was treated with saline, while the Model group (saline) and LJ group (426 mg/kg) had induced traumatic injury. All rats were treated with once by daily oral gavage for one week. The biochemical indices and protein expression were measured. Herein, 79 constituents were identified in LJ, which were effective in elevating body weight, food consumption, water intake, and urinary excretion volume, as well as in ameliorating traumatic muscle tissues in model rats. In addition, LJ prominently decreased the contents of plasma viscosity, platelet aggregation rate, thrombin time, prothrombin time, activated partial thromboplastin time, fibrinogen, thromboxane B2 (TXB2), TXB2/6-keto-prostaglandin F1α (6-keto-PGF1α), urokinase-type plasminogen activator (u-PA), plasminogen activator inhibitor 1 (PAI-1), PAI-1/tissue-type PA (t-PA), and PAI-1/u-PA, while significantly increasing antithrombin III, 6-keto-PGF1α, and t-PA contents. Furthermore, LJ notably inhibited tumor necrosis factor alpha, interleukin 6 (IL-6), IL-8, angiotensin II, antidiuretic hormone, aldosterone, aquaporin 1 (AQP1), AQP2, and AQP3 levels, and markedly elevating IL-10 and natriuretic peptide levels. Finally, LJ markedly reduced the protein expression of AQP1, AQP2, and AQP3 compared to the model group. Collectively, LJ possessed prominent activating blood circulation, anti-inflammatory, and diuretic effects, thus supporting the clinical application of L. japonicus.

Amelioration of chronic prostatitis by fractions of Mongolian medicine Hosta plantaginea flowers via inhibition of NF-κB, MAPKs, JAK-STAT, and PI3K-Akt signaling pathways in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Hosta plantaginea (Lam.) Aschers flower is an important Mongolian medicine beneficial in the treatment of chronic prostatitis (CP) in the absence of scientific evidence. AIM OF THE STUDY: The aim of this study was to reveal the therapeutical effects and potential mechanisms of H. plantaginea flowers extract (HP) and its different polarity fractions (HPA∼D) on autoimmune CP (ACP) model rats. MATERIALS AND METHODS: Sprague-Dawley male rats were randomly assigned to 13 groups (n = 6/group). Except the sham group, all rats were injected with a mixture of prostate antigen and complete Freund's adjuvant on days 0, 7, and 21 to establish ACP model rats. Afterwards, ACP model rats were orally gavaged with HP or HPA∼D (1 and 4 g/kg of raw herbal material) or positive drug (Prostat, 200 mg/kg) daily from day 21 to day 50 for 30 days, while the sham and model groups were treated simultaneously with isopyknic of 0.3% sodium carboxymethyl cellulose. Histopathological analysis, biochemical parameters, and protein expression of prostate tissues were investigated. RESULTS: In comparison with the model group, all fraction groups experienced improved CP effects, including restored body weight, reduced prostate gland edema and prostate index, decreased prostatic leukocytes, increased prostatic lecithin bodies, and alleviated histopathological damage to prostate tissue. Furthermore, all fraction groups markedly inhibited the phosphorylated protein of nuclear factor kappa-B p65 (NF-κB p65), NF-κB inhibitor alpha (IκBα), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (Erk), just another kinase 1 (JAK1), signal transducer and activator of transcription 3 (STAT3), phosphoinositide 3-kinase (PI3K), and protein kinase B (Akt) than the model group. CONCLUSION: All fractions of HP exerted significant anti-CP effects by inhibiting NF-κB, MAPKs, JAK-STAT and PI3K-Akt pathways in ACP model rats. These findings provide scientific evidence that H. plantaginea flowers can be used as a pivotal Mongolian medicine in clinical applications for the treatment of CP.

Kuqin ameliorates Lipopolysaccharide-induced acute lung injury by regulating indoleamine 2,3-dioxygenase 1 and Akkermansia muciniphila.

Scutellariae radix (SR) has been proven to be highly effective in treating inflammation because of its superior medicinal properties. The two main commercial specifications of SR are Kuqin (KQ) and Ziqin (ZQ). According to traditional Chinese medicine theories, KQ has a better effect than ZQ on dispelling upper energizer lung damp heat, however, its mechanism of action is not known. Thus, this study investigated the role of KQ-induced alterations in endogenous metabolites and gut microbiota in regulating LPS-induced acute lung injury (ALI). KQ treatment ameliorated lung injury more effectively than ZQ and demonstrated satisfactory organ protection properties. KQ treatment reversed the tryptophan metabolite abnormalities in ALI and reshaped the composition of gut microbial communities. Additionally, the abundance of the enriched Akkermansia muciniphila was significantly and inversely correlated with the rate-limiting enzyme of the tryptophan/kynurenine pathway, indoleamine 2,3-dioxygenase 1 (IDO1) activity (p = 0.0214, R2 =0.7712). Furthermore, the beneficial and causative effects of A. muciniphila were confirmed by antibiotic and microbial intervention experiments. Live and pasteurized A. muciniphila, both supplements could ameliorate the inflammatory response and down-regulate IDO1 expression, thereby restoring tryptophan metabolic imbalance. In conclusion, the current study demonstrated for the first time that KQ may act on the A. muciniphila abundance, regulate IDO1 activity, and thus ameliorate ALI. Interestingly, A. muciniphila supplementation could be a promising therapeutic option for lung diseases through the gut-lung axis.

Revealing the Active Constituents and Mechanisms of Semiliquidambar cathayensis Chang Roots against Rheumatoid Arthritis through Network Pharmacology, Molecular Docking, and in Vivo Experiment.

Semiliquidambar cathayensis Chang roots (SC) are traditional Chinese medicine for treating rheumatoid arthritis (RA). However, the effect and potential mechanism of SC remain unclear. This study aims to reveal the anti-RA constituents and mechanisms of SC based on network pharmacology, molecular docking, and adjuvant-induced arthritis (AIA) model rat experiment. In this work, 9 potential active constituents, including kaempferol, quercetin, naringenin, paeoniflorin, catechin, fraxin, gentianin, hesperetin, and ellagic acid 3,3',4-trimethyl ether, in SC crossed 65 target genes of RA. In addition, 28 core targets were enriched in inflammation and others, among which interleukin-17 (IL-17) and tumor necrosis factor (TNF) were the major targets. The binding of bio-constituents with IL-17 and TNF were performed using molecular docking. Rat experiment demonstrated that the extract of SC restored body weight loss, reduced arthritis score and the indices of thymus and spleen, alleviated ankle joint histopathology, decreased the levels of rheumatoid factor (RF), C-reactive protein (CRP), IL-17, TNF-α, IL-1ß, IL-6, cyclooxygenase-2 (COX-2), 5-lipoxygenase (5-LOX), and matrix metalloproteinase-2 (MMP-2), whereas elevated the levels of IL-4 and IL-10. Collectively, it was the first time to comprehensively reveal the anti-RA efficacy and mechanism of SC via suppressing the inflammatory pathway based on network pharmacology, molecular docking, and experimental verification, which provide chemical and pharmacological evidences for the clinical application of SC.