Aqueous extract of Epimedium sagittatum (Sieb. et Zucc.) Maxim. induces liver injury in mice via pyroptosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Epimedium sagittatum (Sieb. et Zucc.) Maxim. has been used traditionally in Asia. It can dispel wind and cold, tonify the kidney, and strengthen bones and tendons. However, adverse effects of E. sagittatum have been reported, and the underlying mechanisms remain unclear. AIM OF THE STUDY: This study aimed to investigate liver injury caused by an aqueous extract of E. sagittatum in Institute of Cancer Research (ICR) mice and explore its potential mechanisms. MATERIALS AND METHODS: Dried E. sagittatum leaves were decocted in water to prepare aqueous extracts for ultra-high performance liquid chromatography analysis. Mice were administered an aqueous extract of E. sagittatum equivalent to either 3 g raw E. sagittatum/kg or 10 g raw E. sagittatum/kg once daily via intragastric injection for three months. The liver weights and levels of the serum biochemical parameters including alanine transaminase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), total bilirubin (TBIL), and alkaline phosphatase were measured. Hematoxylin-eosin staining was performed for histopathology. Apoptosis was detected using the TUNEL apoptosis assay kit. IL-1ß was detected using ELISA kits. Proteomics was used to identify the differentially expressed proteins. Western blot analysis was performed to determine the levels of proteins significantly affected by the aqueous extract of E. sagittatum. RESULTS: E. sagittatum treatment increased the liver weights and liver coefficients, and ALT and AST levels significantly increased (p < 0.05). A high dose of E. sagittatum significantly increased LDH and TBIL levels (p < 0.05). Ruptured cell membranes and multiple sites of inflammatory cell infiltration were also observed. No evidence of apoptosis was observed. IL-1ß levels were significantly increased (p < 0.05). The expressions of PIK3R1, p-MAP2K4, p-Jun N-terminal kinase (JNK)/JNK, p-c-Jun, VDAC2, Bax, and CYC were upregulated, whereas that of Bcl-2 was inhibited by E. sagittatum. The expression of cleaved caspase-1 was significantly increased; however, its effects on GSDMD and GSDMD-N were significantly decreased. The expression levels of cleaved caspase-3 and its effector proteins GSDME and GSDME-N significantly increased. CONCLUSIONS: Our results suggest that the aqueous extract of E. sagittatum induces liver injury in ICR mice after three months of intragastric injection via inflammatory pyroptosis.

Investigating the Mechanism of Chinese Medicine Formula AACO Against Chronic Heart Failure by Network Pharmacology Analysis and Experimental Validation.

Background: A traditional Chinese medicine (TCM) formula, containing Astragalus membranaceus (Fisch.) Bunge, Aconitum wilsonii Stapf ex Veitch, Curcuma longa L., and Radix ophiopogonis (AACO), has therapeutic value for the treatment of chronic heart failure (CHF). Objective: This study intends to explore the pharmacological mechanism underlying the activity of the AACO formula against CHF. Materials and Methods: Using the TCM Systems Pharmacology database and Bioinformatics Analysis Tool for Molecular Mechanism of TCM, the active ingredients contained in the herbs of the AACO formula were screened. Meanwhile, the target genes related to these active ingredients were identified and genes correlated with CHF were screened. Protein-protein interaction networks were built to elucidate the relationships between the AACO formula and CHF. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) signal pathway enrichment analysis were carried out using the DAVID database. A "drug-component-target-disease" network was constructed with Cytoscape 3.7.0. The therapeutic effect of the AACO formula was proven by hemodynamic study, echocardiography evaluation, and histological analysis in transverse aortic constriction-induced CHF mice and was validated in vitro. Results: A total of 105 active ingredients and 1026 related targets were screened and identified, and 240 related targets overlapping with CHF were selected. According to GO analysis, the enriched genes participated in gene expression and cardiac contraction regulation by Ca2+ regulation. From KEGG analysis, the calcium axis was identified as one of the main mechanisms through which the AACO formula exerts an anti-CHF effect. AACO was validated to significantly improve cardiac diastolic and systolic functions in vivo via an increase in the rate of Ca2+ reuptake of the myocardial sarcoplasmic reticulum and improved myocardial contractility in vitro. Conclusions: Network pharmacology is a convenient method to study the complex pharmacological mechanisms of TCM. The calcium axis likely participates in the anti-CHF mechanism of AACO.

Panax quinquefolius saponins combined with dual antiplatelet therapy enhanced platelet inhibition with alleviated gastric injury via regulating eicosanoids metabolism.

BACKGROUND: Panax quinquefolius saponin (PQS) was shown beneficial against platelet adhesion and for gastroprotection. This study aimed to investigate the integrated efficacy of PQS with dual antiplatelet therapy (DAPT) on platelet aggregation, myocardial infarction (MI) expansion and gastric injury in a rat model of acute MI (AMI) and to explore the mechanism regarding arachidonic acid (AA)-derived eicosanoids metabolism. METHODS: Wistar rats were subjected to left coronary artery occlusion to induce AMI model followed by treatment with DAPT, PQS or the combined therapy. Platelet aggregation was measured by light transmission aggregometry. Infarct size, myocardial histopathology was evaluated by TTC and H&E staining, respectively. Gastric mucosal injury was examined by scanning electron microscope (SEM). A comprehensive eicosanoids profile in plasma and gastric mucosa was characterized by liquid chromatography-mass spectrometer-based lipidomic analysis. RESULTS: PQS+DAPT further decreased platelet aggregation, lessened infarction and attenuated cardiac injury compared with DAPT. Plasma lipidomic analysis revealed significantly increased synthesis of epoxyeicosatrienoic acid (EET) and prostaglandin (PG) I2 (potent inhibitors for platelet adhesion and aggregation) while markedly decreased thromboxane (TX) A2 (an agonist for platelet activation and thrombosis) by PQS+DAPT, relative to DAPT. DAPT induced overt gastric mucosal damage, which was attenuated by PQS co-administration. Mucosal gastroprotective PGs (PGE2, PGD2 and PGI2) were consistently increased after supplementation of PQS+DAPT. CONCLUSIONS: Collectively, PQS+DAPT showed synergistic effect in platelet inhibition with ameliorated MI expansion partially through upregulation of AA/EET and AA/PGI2 synthesis while suppression of AA/TXA2 metabolism. PQS attenuated DAPT-induced gastric injury, which was mechanistically linked to increased mucosal PG production.

A dual-nanozyme-loaded black phosphorus multifunctional therapeutic platform for combined photothermal/photodynamic/starvation cancer therapy.

The typical hypoxia of tumor microenvironments seriously affects the efficacy of starvation therapy (ST) and photodynamic therapy (PDT). Therefore, it is of great significance to prepare a multimodal combined therapy nanocomposite with the ability to relieve tumor hypoxia. It is an effective method to release oxygen into the tumor microenvironment using a nanoenzyme. In this work, two-dimensional black phosphorus nanosheets (BPNSs) were used as photothermal reagents and photosensitized agents due to the unique physical properties and also used as nano-carriers for the in situ reduction deposition of Au nanoparticles and fragmented MnO2 coatings. Finally, polyethylene glycol was introduced to obtain the nanocomposite BP@Au@MnO2-PEG (i.e., AMGP). In the tumor microenvironment, MnO2 can catalyze endogenous H2O2 to produce oxygen, improving the effect of ST and PDT. H2O2 generated via the catalytic oxidation of glucose by Au nanoparticles can continue to be used as the substrate catalyzed by MnO2. Meanwhile, black phosphorus nanosheets have excellent photothermal properties for photothermal treatment. The results of in vitro and in vivo experiments indicated that AMGP nanocomposites have good combined antitumor efficacies and biosafety.

A novel green phosphorus-containing flame retardant finishing on polysaccharide-modified polyamide 66 fabric for improving hydrophilicity and durability.

Rising concerns about the toxic effects and environmental issues associated with various fireproof treatments on textiles have led to a demand for "green" materials. Chitosan (CS) is an amino polysaccharide green, recyclable, and non-toxic highly biocompatible biopolymer that consists of multiple hydroxyl groups and has a wide range of applications, including as a flame retardant additive. In this study, an eco-friendly bio-based formaldehyde-free flame retardant containing a higher level of phosphorus and nitrogen in phytic acid ammonia (PAA) was synthesized to amplify the most plentiful green chitosan (CS)-modified polyamide 66 (PA66) fabric surface through a simple pad-dry-cure technique for the improvement of durable flame retardancy with hydrophilicity. The findings revealed that each UV-grafted CS fabric could entirely stop the melt-dripping tendency during the vertical burning (UL-94) test and reached a V-1 rating. Meanwhile, limiting oxygen index (LOI) testing showed a rapid increase from 18.5 % to 24 % for the PA66 control and the PAA-treated (i.e., PA66-g-5CS-PAA) fabric samples, respectively. Moreover, compared to the PA66 control sample, a dramatic decrease in the peak heat release rate (PHRR), fire growth rate (FGR), and total heat release (THR) by approximately over 52 %, 0.63 %, and 19.7 %, respectively, was observed for the PA66-g-5CS-PAA fabric sample. Additionally, this arrangement of PAA catalyzed the charring of grafted CS and acted as a condensed phase flame retardant, resulting in a significant improvement in char yield% in both air and N2 atmospheres for the PA66-g-5CS-PAA fabric sample in TGA. In addition, only the lower grafting ratio of CS with PAA-treated fabric sample (i.e., PA66-g-2CS-PAA) could encourage it to gain its lowest water contact angle of 00, as well as impersonating a positive effect in improving the flame retardant coating durability in washing and sustaining even after 10 home laundering cycles. This phenomenon suggests that an actual hydrophilic and durable flame retardant finishing procedure for polyamide 66 fabrics might be applied with the novel, plentiful, sustainable, and environmentally friendly bio-based green PAA ingredient.

Altered Functional Activity and Functional Connectivity of Seed Regions Based on ALFF Following Acupuncture Treatment in Patients with Stroke Sequelae with Unilateral Limb Numbness.

Objective: Limb numbness is a frequent symptom of post-stroke somatosensory dysfunction, which may be alleviated by non-invasive therapy such as acupuncture. However, the precise mechanism via acupuncture remains unknown. The goal of this study was to investigate how the amplitude of low-frequency fluctuations (ALFF) and functional connectivity (FC) changed between stroke patients with limb numbness and healthy people, as well as how acupuncture might work. Methods: 24 stroke sequelae patients with unilateral limb numbness and 14 matched healthy controls were enrolled in the study. The patients with limb numbness received acupuncture therapy three days a week for four weeks. We mainly assessed the clinical outcomes via the visual analogue scale (VAS). In addition, fMRI data from patients with unilateral limb numbness at baseline and after treatment (4th week) were collected, as well as data from healthy controls at baseline. Results: Compared with the healthy subjects, the patient group demonstrated significantly decreased ALFF in several brain regions, mainly associated with the sensorimotor network (SMN) and default mode network (DMN), including left superior frontal gyrus (SFG), right temporal fusiform cortex (TFC), right middle frontal gyrus (MFG), bilateral middle temporal gyrus (MTG), right putamen (PUT), right precentral gyrus (preCG), right planum polare (PP), and left supplementary motor area (SMA). These regions were chosen as the seeds for investigating the FC alteration induced by acupuncture. Several sensorimotor-related brain regions were activated by acupuncture, and the FC of the left supramarginal gyrus (SMG) with right MTG, as well as brain-stem, cerebellum vermis 9 with right MFG showed enhancement following acupuncture in the patient group, which had a significant correlation with clinical outcomes. Conclusion: Acupuncture treatment may be used to stimulate brain areas associated with somatosensory processing and to strengthen the FC of sensorimotor and cognitive brain networks in order to achieve therapeutic effect.

Clinical Significance of Procalcitonin, Lactic Acid, and Endotoxin Testing for Children With Severe Pneumonia and Sepsis.

Context: Sepsis is a common complication of severe pediatric pneumonia, characterized by difficulty in treatment, a high treatment cost, high morbidity and mortality, and poor prognosis. The levels of three indicators, procalcitonin (PCT), lactic acid (Lac), and endotoxin (ET), can vary greatly in children with severe pneumonia complicated by sepsis. Objective: The study aimed to investigate the clinical significance of PCT, Lac, and ET levels in the serum of children with severe pneumonia complicated by sepsis. Design: The research team performed retrospective study. Setting: The study took place at Nantong First People's Hospital in Nantong, Jiangsu, China. Participants: Participants were 90 children with severe pneumonia complicated by sepsis and 30 children with severe pneumonia only, all of whom had received treatment in the pediatric intensive care unit of the hospital between January 2018 and May 2020. Groups: At baseline, the research team divided the participants into three groups based on their pediatric clinical illness score (PCIS) at 24 h after admission: (1) the extremely critical group-0-70 points (n = 29), (2) the critical group-71-80 points (n = 31), and (3) the noncritical group->80 points (n = 30). The 30 children who had received treatment but who had severe pneumonia only became the control group. Outcome Measures: The research team: (1) measured the serum PCT, Lac, and ET levels for the four groups at baseline, (2) compared those levels by group, (3) compared those levels by clinical outcome, (4) determined the correlation of the three indicators to the PCIS scores, and (5) identified the predictive value of the three indicators. To compare the levels by clinical outcome and to determine the indicators' predictive values, the team divided participants into two groups according to their clinical outcomes on day 28 of the study: (1) 40 children who died became the death group, and (2) 50 children who survived became the survival group. Results: The serum PCT, Lac, and ET levels in the extremely critical group were the highest, followed by the critical group, the noncritical group, and the control group. The serum PCT, Lac, and ET levels had a significant negative correlation with participants' PCIS scores (r = -0.8203 (PCT), -0.6384 (Lac), -0.6412 (ET), P < .05).The area under the curve (AUC) for the PCT level was 0.7732 (95% CI = 0.6214 to 0.9249, P = .0015), for the Lac level was 0.9533 (95% CI = 0.9036 to 1.000, P < .0001), and for the ET level was 0.8694 (95% CI = 0.7622 to 0.9765, P < .0001). These values indicate that all three indicators were significantly predictive regarding participants' prognoses. Conclusions: The serum PCT, Lac, and ET in children with severe pneumonia complicated by sepsis were abnormally high, and the levels of these indicators were significantly negatively correlated with the PCIS scores. PCT, Lac, and ET may be potential indicators for the diagnosis and prognosis assessment of children with severe pneumonia complicated by sepsis.

Remediation of copper and lead contaminated sediments using iron-based granule biochar: mechanisms and enzyme activity.

In recent years, there has been a growing concern about heavy metal contamination in sediments. In this study, iron-based granular biochar (MGB) is prepared to remediate Cu and Pb contaminated sediments. Characterizations via scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) show that the rough surface of MGB with abundant pore structures and a large number of oxygen-containing functional groups that facilitate stabilization of Cu and Pb in sediments. Potential mobility and bioavailability of Cu and Pb are investigated using BCR sequential extraction in the 35 day remediation experiment. The XPS results indicate that FeOOH and C-OH play a crucial part in stabilizing heavy metals. Large affinity of FeOOH for Pb allows it to occupy a proportion in F2 while C-OH is attractive to Cu. Changes of pH, organic matter (OM), and available phosphorus (AP) in sediments after adding MGB as well as the relationship between changes and the stable solidification of Cu and Pb are explored. The stable solidification of heavy metals effectively reduces the available phosphorus in sediments. Magnetic and particle properties of the material are used to reduce the impact of MGB aging on sediment environment and separate it from the remediated sediment. Finally, 3% of MGB significantly enhanced the sediment catalase activity in the biological enzyme activity experiment. All findings indicate that MGB is a green and environmentally friendly sediment remediation material with satisfactory potential in synergistically stabilizing heavy metals and phosphorus.Highlights The complexation of FeOOH with Pb on the surface of MGB fixes it to the reduced stateThe C-OH on the surface of MGB is more attractive to Cu than PbMGB effectively mitigates the release of bioavailable phosphorus from sediments to overlying water.

Sub-chronic toxicity of an aqueous extract of Epimedium sagittatum (Sieb. Et Zucc.) Maxim. in rats.

Epimedium sagittatum (Sieb. et Zucc.) Maxim., a traditional medicinal plant in Asia, is widely used in clinical settings but its safety in vivo is unclear. This study investigated the sub-chronic toxicity of E. sagittatum aqueous extract to rats with a 13-week daily intragastric administration of 7.5, 15, or 30 g/kg. Nine constituents of the aqueous extract were identified by ultra-performance liquid chromatography (UPLC). Organ weights, organ coefficients, serum biochemistry parameters, histopathology, and metabolomic analysis were performed. In female rats, treatment increased the liver, thymus, and adrenal gland coefficients (p < 0.05). Liver, pancreas, and adrenal gland injury were observed. The levels of six metabolites were altered by the treatment (p < 0.05). In male rats, treatment altered liver, heart, and thymus coefficients (p < 0.05) and liver, adrenal gland, and heart injury were observed. The levels of 11 metabolites were altered (p < 0.05). The no-observed-adverse-effect level was not determined but would be below 7.5 g/kg in rats treated for 13 weeks. In female rats, E. sagittatum may injure the liver and pancreas and dysregulate the biosynthesis of phenylalanine, tyrosine, tryptophan, valine, leucine, and isoleucine and the metabolism of phenylalanine. In male rats, the extract may injure the liver and adrenal gland and dysregulate the biosynthesis of valine, leucine, and isoleucine and the metabolism of pyruvate.

Pentacyclic triterpenoids from spikes of Prunella vulgaris L. with thyroid tumour cell cytostatic bioactivities.

Five new triterpenoids, including four ursane types (1-4) and one oleanane type (5), together with 15 known ursane types pentacyclic triterpenoids (6-20) were isolated from the fruit spikes of Prunella vulgaris L., a traditional Chinese herbal medicine. Their structures were elucidated based on IR, HR-ESI-MS, and NMR spectroscopic data. The SW579 cell line was used to evaluate anti-thyroid cancer activities of (1-20). The results indicated that (7-9), (16), and (19) exhibited apparent inhibitory activity with IC50 values of 25.73-71.41 µM (cisplatin as positive control, IC50 14.49 ± 0.97 µM). Network pharmacology and molecular docking were also used for the prediction of the synergistic actions and the underlying mechanisms. Accordingly, four potential targets have been characterized.

Dryness weakens the positive effects of plant and fungal ß diversities on above- and belowground biomass.

Plant and microbial diversity are key to determine ecosystem functioning. Despite the well-known role of local-scale α diversity in affecting vegetation biomass, the effects of community heterogeneity (ß diversity) of plants and soil microbes on above- and belowground biomass (AGB and BGB) across contrasting environments still remain unclear. Here, we conducted a dryness-gradient transect survey over 3000 km across grasslands on the Tibetan Plateau. We found that plant ß diversity was more dominant than α diversity in maintaining higher levels of AGB, while soil fungal ß diversity was the key driver in enhancing BGB. However, these positive effects of plant and microbial ß diversity on AGB and BGB were strongly weakened by increasing climatic dryness, mainly because higher soil available phosphorus caused by increasing dryness reduced both plant and soil fungal ß diversities. Overall, these new findings highlight the critical role of above- and belowground ß diversity in sustaining grassland biomass, raising our awareness to the ecological risks of large-scale biotic homogenization under future climate change.

A natural compound hyperoside targets Salmonella Typhimurium T3SS needle protein InvG.

The antimicrobial actions of natural compounds derived from medicinal plants have been well documented. However, their detailed mechanisms underlying the action against microorganisms remain largely unexplored. Salmonella enterica is a common pathogen causing both gastrointestinal and systemic diseases. In Salmonella enterica, the type III secretion system (T3SS) is employed to export secreted effectors directly to the cytoplasm of host cells. Using a SipA-ß-lactamase reporter, we found that hyperoside (HYP) inhibited the activity of Salmonella T3SS needle protein InvG, prevented damage to host cells and protected mice against Salmonella enterica serovar Typhimurium. It was also observed that HYP binds to InvG directly through hydrogen-bridged cations and hydrophobic interactions. The unique mechanism of antibacterial action of HYP suggested that it could be used as a potentially effective candidate for future antimicrobial regimens.

Oxygen-deficient titanium dioxide-loaded black phosphorus nanosheets for synergistic photothermal and sonodynamic cancer therapy.

Malignant tumors, particularly those located in deep tissues, have always been a grievous threat to human health. Sonodynamic therapy (SDT) has recently attracted great attention due to deep tissue penetration. However, the lack of effective sonosensitizers and the poor therapeutic efficacy severely limit their wider use. Herein, dual-functionalized black phosphorus nanosheets (BP@PEI-PEG, i.e., PPBP) integrating black oxygen-deficient titanium dioxide particles (B-TiO2) were successfully constructed (PPBP-B-TiO2) for synergistic photothermal (PTT)/sonodynamic therapy. In these nanocomposites, black titanium dioxide can enhance the separation of electrons (e-) and holes (h+) due to the oxygen-deficient structure and significantly improves the production of reactive oxygen species (ROS) for SDT, while the BP nanosheets endow the nanocomposites with a higher photothermal conversion capability for photothermal therapy (η = 44.1%) which can prolong the blood circulation and improve the O2 supply. In vivo experiments prove that PPBP-B-TiO2 nanocomposites exhibited outstanding tumor inhibition efficacy and excellent biocompatibility. This work provides a prospective platform for combined photothermal/sonodynamic cancer therapy.

Hemin-loaded black phosphorus-based nanosystem for enhanced photodynamic therapy and a synergistic photothermally/photodynamically activated inflammatory immune response.

The biocompatible nanosystem integrating hemin into black phosphorus nanosheets was ingeniously constructed through the easy modified strategy. Taking advantage of the enhanced permeability and retention (EPR) effect, the designed nanosystem could accumulate into the tumor location, leading to attractive cytotoxicity through the enhanced photodynamic therapy (PDT) ascribing to the catalytic oxygen supply and GSH depletion of hemin. Simultaneously, combining PDT and photothermal therapy (PTT) showed an apparent promotion in anti-tumor effect. Moreover, inflammatory response and immune activation amplified anti-tumor effect, which could compensate limitations of exogenous therapy (i.e., limited tissue depth and intensity-dependent curation effect) and potentiate the efficiency of the endogenous immune-activating behavior. Especially, the designed nanosystem degraded followed by being metabolized in the blood circulation. By and large, this constructed nanosystem provides the new insight into designing biocompatible nanomaterials and paves the ideal way for anti-tumor therapy. STATEMENT OF SIGNIFICANCE: Biocompatible nanomaterials-based synergistic tumor therapy offers the potential application prospect. Taking advantage of degradable black phosphorus, the nanosystem integrating hemin into black phosphorus for the enhanced photodynamic therapy and synergistic photothermal-photodynamic activating inflammation-immune response was developed and the results demonstrate that tumor growth was inhibited followed by activating inflammatory factors and leading to satisfactory immune response.

Covalent grafting diazotized black phosphorus with ferrocene oligomer towards smoke suppression and toxicity reduction.

In comparison with the thermal hazard of polymers, noxious smoke and gas produced by the combustion of polymers make the environment self-purification a huge challenge. As a new type of a highly effective flame retardant, black phosphorus (BP) can effectively decrease the thermal hazard of polymers, but its performances in smoke suppression and toxicity reduction are unsatisfactory. In this article, a method of covalently grafting diazotized BP with a ferrocene oligomer was applied to promote the smoke suppression and toxicity reduction efficiency of BP. In our work, the BP-NH nanomaterials with a mass of amino groups on the surface were acquired by diazotizing the BP. Then, the BP-Fe was obtained by covalently grafting the ferrocene chloride salt and nitrogen-containing heterocycles on the surface of BP. The smoke production rate (SPR) and total smoke production (TSP) values of the epoxy resin (EP) decreased by 49.8% and 52.5% with the addition of 2 wt% BP-Fe, respectively. In comparison with previous studies, this work was far more effective than the previous work in smoke suppression and flame retardant. The release of toxic gases (CO and HCN) and volatile organic compounds in the EP was also effectively inhibited at the same time. In addition, the storage modulus and tensile strength of nanocomposites increased by 35.1% and 27.2% with the addition of 1 wt% BP-Fe. This work also provides a new idea on how to simultaneously strengthen the toxic smoke suppression, mechanical properties, and flame retardant of polymer materials.

Inhibitory Effects of Mongolian Medicine Yihe-Tang on Continuous Darkness Induced Liver Steatosis in Zebrafish.

The constant dark induction (DD) causes lipid degeneration and nonalcoholic fatty liver disease (NAFLD) in zebrafish, which might be closely related to the imbalance of gut microbiota and require in-depth study. In this study, a total of 144 zebrafish were divided into four groups, including the control group, Yihe-Tang group, constant dark group, and constant dark + Yihe-Tang group, and were treated with constant darkness (except control and Yihe-Tang groups) for 21 days. The bodyweights of zebrafish were recorded after 8 d, 15 d, and 22 d. The sequencing analysis of gut microbiota, detection of liver histopathological changes, and comparison of lipid metabolism-related gene expression levels were performed on the 22nd day of the experiment. The results showed that the Yihe-Tang could inhibit the constant dark-induced increase in zebrafish weight and liver steatosis. As compared to the control group, the dark treatment could alter the composition of gut microbiota in zebrafish, increase the relative abundance of harmful bacteria, and decrease the Cetobacterium and Bacteroides to Firmicutes ratio in the intestines. The abundance of Proteobacteria in the constant dark + Yihe-Tang group was close to that in the control group and that of Fusobacteria and Cetobacterium increased, especially the Cetobacterium, which increased significantly. The constant dark treatment caused an abnormal expression of liver lipid-related genes, inhibited lipid metabolism, and promoted fat accumulation. However, the Yihe-Tang could restore these changes to the level of the control group. This study indicated that Yihe-Tang could restore the constant dark-induced liver lipid degeneration. We hypothesized that Cetobacterium could significantly inhibit steatosis.

Cardioprotective effects of gypenoside XVII against ischemia/reperfusion injury: Role of endoplasmic reticulum stress, autophagy, and mitochondrial fusion fission balance.

Gypenoside XVII (GP-17), a tetracyclic triterpene saponin isolated from the functional food Gynostemma pentaphyllum, has been demonstrated protective effects against cerebrovascular and cardiovascular diseases on multiple disease models. In this study, we established a myocardial infarction (MI) model by ligating the left anterior descending coronary artery, and explored whether GP-17 prevent myocardial ischemia/reperfusion (I/R) injuries in mice. Compared with the I/R group, GP-17 significantly improved the cardiac function, reduced the MI, decreased myocardial pathology, activated superoxide dismutase and catalase, and reduced the content of lactate dehydrogenase, creatine kinase, malondialdehyde, and inflammatory factor. The proteomic analysis showed multiple differential proteins between the GP-17 and I/R groups enriched in endoplasmic reticulum and mitochondria. Western-Blot showed that GP-17 significantly decreased the expression of GRP78, ATF6, CHOP, and phosphorylation of PERK, indicating the inhibition of ERS. GP-17 inhibited the expression of ATG5, LC3A/B, and BAX, illustrating the suppression of autophagy and apoptosis. Moreover, both GP-17 and 4-PBA could improve the downregulated Mfn2, meaning that inhibition of ERS regulated the mitochondrial fusion fission balance, thus protected the function of mitochondria. In conclusion, we found that GP-17 prevented against myocardial I/R injury by inhibit ERS-induced cell apoptosis, autophagy, oxidative stress, and mitochondrial division.

In Silico Identification and Mechanism Exploration of Active Ingredients against Stroke from An-Gong-Niu-Huang-Wan (AGNHW) Formula.

An-Gong-Niu-Huang-Wan (AGNHW) is a well-known formula for treating cerebrovascular diseases, with roles including clearing away heat, detoxification, and wake-up consciousness. In recent years, AGNHW has been commonly used for the treatment of ischemic stroke, but the mechanism by which AGNHW relieves stroke has not been clearly elucidated. In the current study, we developed a multiple systems pharmacology-based framework to identify the potential antistroke ingredients in AGNHW and explore the underlying mechanisms of action (MOA) of AGNHW against stroke from a holistic perspective. Specifically, we performed a network-based method to identify the potential antistroke ingredients in AGNHW by integrating the drug-target network and stroke-associated genes. Furthermore, the oxygen-glucose deprivation/reoxygenation (OGD/R) model was used to validate the anti-inflammatory effects of the key ingredients by determining the levels of inflammatory cytokines, including interleukin (IL)-6, IL-1ß, and tumor necrosis factor (TNF)-α. The antiapoptotic effects of the key ingredients were also confirmed in vitro. Integrated pathway analysis of AGNHW revealed that it might regulate three biological signaling pathways, including IL-17, TNF, and PI3K-AKT, to play a protective role in stroke. Moreover, 30 key antistroke ingredients in AGNHW were identified via network-based in silico prediction and were confirmed to have known neuroprotective effects. After drug-like property evaluation and pharmacological validation in vitro, scutellarein (SCU) and caprylic acid (CA) were selected for further antistroke investigation. Finally, systems pharmacology-based analysis of CA and SCU indicated that they might exert antistroke effects via the apoptotic signaling pathway and inflammatory response, which was further validated in an in vitro stroke model. Overall, the current study proposes an integrative systems pharmacology approach to identify antistroke ingredients and demonstrate the underlying pharmacological MOA of AGNHW in stroke, which provides an alternative strategy to investigate novel traditional Chinese medicine formulas for complex diseases.

The chloroplast genome sequence and phylogenetic analysis of Apocynum venetum L.

Apocynum venetum L. (Apocynaceae) is valuable for its medicinal compounds and fiber content. Native A. venetum populations are threatened and require protection. Wild A. venetum resources are limited relative to market demand and a poor understanding of the composition of A. venetum at the molecular level. The chloroplast genome contains genetic markers for phylogenetic analysis, genetic diversity evaluation, and molecular identification. In this study, the entire genome of the A. venetum chloroplast was sequenced and analyzed. The A. venetum cp genome is 150,878 bp, with a pair of inverted repeat regions (IRA and IRB). Each inverted repeat region is 25,810 bp, which consist of large (LSC, 81,951 bp) and small (SSC, 17,307 bp) single copy areas. The genome-wide GC content was 38.35%, LSC made up 36.49%, SSC made up 32.41%, and IR made up 43.3%. The A. venetum chloroplast genome encodes 131 genes, including 86 protein-coding genes, eight ribosomal RNA genes, and 37 transfer RNA genes. This study identified the unique characteristics of the A. venetum chloroplast genome, which will help formulate effective conservation and management strategies as well as molecular identification approaches for this important medicinal plant.

Anti-depressive effects of Jiao-Tai-Wan on CORT-induced depression in mice by inhibiting inflammation and microglia activation.

ETHNOPHARMACOLOGICAL RELEVANCE: Jiao-Tai-Wan (JTW) is a very famous traditional Chinese medicine formula for the treatment of psychiatric disorders, especially in anxiety, insomnia and depression. However, its molecular mechanism of treatment remains indistinct. AIM OF THE STUDY: We aimed to reveal the action mechanism of JTW on anti-depression via inhibiting microglia activation and pro-inflammatory response both in vivo and in vitro. MATERIAL AND METHODS: The corticosterone (CORT)-induced depression mouse model was used to evaluate the therapeutic efficacy of JTW. Behavioral tests (open field, elevated plus maze, tail suspension and forced swim test) were conducted to evaluate the effect of JTW on depressive-like behaviors. The levels of inflammatory factors and the concentration of neurotransmitters were detected by RT-qPCR or ELISA assays. Then three hippocampal tissue samples per group (Control, CORT, and JTW group) were sent for RNA sequencing (RNA-seq). Transcriptomics data analysis was used to screen the key potential therapeutic targets and signaling pathways of JTW. Based on 8 bioactive species of JTW by our previous study using High-performance liquid chromatography (HPLC) analysis, molecular docking analyses were used to predict the interaction of JTW-derived compounds and depression targets. Finally, the results of transcriptome and molecular docking analyses were combined to verify the targets, key pathways, and efficacy of JTW treatment in vivo and vitro. RESULTS: JTW ameliorated CORT-induced depressive-like behaviors, neuronal damage and enhanced the levels of monoamine neurotransmitters in the serum of mice. JTW also inhibited CORT-induced inflammatory activation of microglia and decreased the serum levels of interleukin- 6(IL-6) and interleukin- 1ß (IL-1ß) in vivo. Transcriptomic data analysis showed there were 10 key driver analysis (KDA) genes with the strongest correlation which JTW regulated in depression mice. Molecular docking analysis displayed bioactive compound Magnoflorine had the strongest binding force to the key gene colony-stimulating factor 1 receptor (CSF1R), which is the signaling microglia dependent upon for their survival. Meanwhile, CSF1R staining showed it was consistent with inflammatory activation of microglia. Our vitro experiment also showed JTW and CSF1R inhibitor significantly reduced lipopolysaccharide (LPS)/interferon-gamma (IFNÉ£)-induced inflammatory activation response in macrophage cells. CONCLUSIONS: Our study suggests that JTW might ameliorate CORT-induced neuronal damage in depression mice by inhibiting CSF1R mediated microglia activation and pro-inflammatory response.