Determining the mechanism of action of the Qishan formula against lung adenocarcinoma by integration of network pharmacology, molecular docking, and proteomics.

BACKGROUND: Lung adenocarcinoma (LUAD) is the main pathological type of lung cancer. Qishan formula (QSF) is reportedly efficacious against LUAD. However, its mechanisms of action currently remain elusive. Therefore, network pharmacology, molecular docking techniques and proteomics were used to verify the potential pharmacological effects of QSF in the treatment of LUAD. METHODS: The active ingredients and potential targets of QSF were obtained from the TCMSP, chemical source network and construct a drug-component-target networks using Cytoscape v3.7.2. Data for disease targets were obtained from 5 databases: TCGA, OMIM, DrugBank, DisGeNET, and GeneCards. Drug disease cross targets were used to construct protein-protein interaction networks for selecting the core targets using the STRING database and enrichment pathway networks using the DAVID database. Finally, TMT quantitative proteomics was used to identify the possible core targets and action pathways. Molecular docking to verify the affinity between components and targets. RESULTS: Network pharmacology identified core components of QSF against LUAD included baicalein, methylophiopogonone B, quercetin, kaempferol, isorhamnetin, and luteolin, which can act on 10 key targets (SRC, TP53, PIK3R1, MAPK3, STAT3, MAKP1, HSP90AA1, PIK3CA, HRAS, and AKT1). QSF might play a therapeutic role in LUAD by regulating biological processes such as signal transduction, protein phosphorylation, cell proliferation, and apoptosis, as well as the PI3K/AKT, MAPK, FoxO, and other signaling pathways. Proteomics identified 207 differentially expressed proteins, and by integrating with network pharmacology and molecular docking results we found that 6 core components of QSF may target TP53 against LUAD through the PI3K/AKT signaling pathway. CONCLUSION: QSF is a multitarget recipe potentially exerting pleiotropic effects in LUAD.

Antioxidant Effects of Roasted Licorice in a Zebrafish Model and Its Mechanisms.

Licorice (Gan-Cao, licorice) is a natural antioxidant and roasted licorice is the most common processing specification used in traditional Chinese medicine prescriptions. Traditional Chinese medicine theory deems that the honey-roasting process can promote the efficacy of licorice, including tonifying the spleen and augmenting "Qi" (energy). The antioxidant activity and mechanisms underlying roasted licorice have not yet been reported. In this study, we found that roasted licorice could relieve the oxidative stress injury induced by metronidazole (MTZ) and could restrain the production of excessive reactive oxygen species (ROS) induced by 2,2'-azobis (2-methylpropionamidine) dihydrochloride (AAPH) in a zebrafish model. It was further found that roasted licorice could exert its oxidative activity by upregulating the expression of key genes such as heme oxygenase 1 (HO-1), NAD(P)H quinone dehydrogenase 1 (NQO1), glutamate-cysteine ligase modifier subunit (GCLM), and glutamate-cysteine ligase catalytic subunit (GCLC) in the nuclear factor erythroid 2-related factor 2 (NRF2) signaling pathway both in vivo and in vitro. Furthermore, consistent results were obtained showing that rat serum containing roasted licorice was estimated to reduce cell apoptosis induced by H2O2. Then, the UHPLC-Q-Exactive Orbitrap MS analysis results elucidated the chemical composition of rat plasma containing roasted licorice extracts, including ten prototype chemical components and five metabolic components. Among them, six compounds were found to have binding activity with Kelch-like ECH-associated protein 1 (KEAP1), which plays a crucial role in the transcriptional activity of NRF2, using a molecular docking simulation. The results also showed that liquiritigenin had the strongest binding ability with KEAP1. Immunofluorescence further confirmed that liquiritigenin could induce the nuclear translocation of NRF2. In summary, this study provides a better understanding of the antioxidant effect and mechanisms of roasted licorice, and lays a theoretical foundation for the development of a potential antioxidant for use in clinical practice.

Traditional Chinese medicine Jianpi therapy in exercise-induced fatigue: A protocol for systematic review and meta-analysis.

BACKGROUND: Exercise-induced fatigue (EIF) is a common occurrence in sports competition and training. It may cause trouble to athletes' motor skill execution and cognition. Although traditional Chinese medicine Jianpi therapy has been commonly used for EIF management, relevant evidence on the effectiveness and safety of Jianpi therapy is still unclear. METHODS: Databases including PubMed, Embase, Web of Science, the Cochrane Library, SinoMed, China Science and Technology Journal Database (VIP), China National Knowledge Infrastructure (CNKI), and Wanfang will be searched for relevant randomized controlled trials from databases from 2000 to 2021. Randomized controlled trials related to traditional Chinese medicine Jianpi therapy in the treatment and management of EIF will be included. Systematic review and meta-analysis of the data will be performed in RevMan 5.3 according to the Preferred Reporting Items of Systematic Reviews and Meta-Analysis (PRISMA) guidelines. Two authors independently performed the literature searching, data extraction, and quality evaluation. Risk of bias was assessed using the Cochrane Risk of Bias Tool for randomized clinical trials. RESULTS: This systematic review and meta-analysis will summarize the latest evidence for traditional Chinese medicine Jianpi therapy in EIF. The results will be submitted to a peer-reviewed journal once completed. CONCLUSION: The conclusion of our research will provide evidence to support traditional Chinese medicine Jianpi therapy as an effective intervention for patients with EIF.OSF Registration DOI: 10.17605/OSF.IO/NRKX4.

Using sediment resuspension to immobilize sedimentary phosphorus.

The response of the transformation of internal phosphorus (P) to resuspended sediment was investigated in the sediment-water system under different disturbance intensity. The sediments and overlying water were collected from Taihu Lake, a typical shallow lake. The concentrations of particulate P (PP) and dissolved inorganic P (DIP) in the water phase and algal available P (AAP) and P fractions in the sediments and suspended particle characteristics were evaluated in laboratory-simulated experiments. The results show that dissolved oxygen (DO) in the overlying water increased continuously and pH decreased slightly under sediment resuspension. The concentration of ammonia nitrogen (NH4+-N) showed a distinct decline, indicating that sediment resuspension promotes the penetration of DO into the sediments. It was also favor of the formation of metal oxides and hydroxides, inducing soluble amorphous metal compounds oxidized to insoluble crystalline metal compounds under disturbance condition. This resulted in the increase of refractory P in sediment compared with the static conditions. Sediment resuspension is beneficial to long-term P retention. This can be confirmed by the increase of maximum P adsorption amount (Qmax) and the decrease of the degree of P saturation (DPS) and equilibrium P concentration (EPC0). This is the main explanation of DIP decrease in the overlying water. It is indicating that sediment resuspension not only improves the redox environment in the sediment-water system but also enhances P retention capacity.

The effect of microenvironment in the sediment on phosphorus immobilization under capping with ACPM and Phoslock®.

Currently, in situ capping is a typical popular geoengineering method for eutrophication control. It is crucial to better understand the effect of microenvironment change due to capping, such as amended calcium peroxide material (ACPM) and Phoslock®, on phosphorus (P) adsorption and immobilization under the addition of external P. The microenvironment in sediment was presented by the concentration of O2, NH4+, and Fe2+ and microbial activity. The P removal and immobilization were also analyzed. The results show that the stronger oxidation in the microenvironment under the capping with ACPM was due to the higher reduction of NH4+ and Fe2+ and the higher increase of microbial activity, compared to Phoslock®. Although, under the capping of ACPM, less amount of external P was removed and there was a faster release of sedimentary P, compared to Phoslock®, ACPM improved the transformation of P from mobile P fractions to inert P fractions. In addition, sedimentary P under the capping of ACPM presents less release than that under the capping of Phoslock® during the anaerobic incubation. However, the settlement of suspended solids decreased the function of capping. All these results indicated that the mechanism of P removal and immobilization was different under the capping of ACPM and Phoslock®.

[Environmental Significance of Phosphorus Fractions of Phytoplankton-and Macrophyte-Dominated Zones in Taihu Lake].

It is of great importance to study the environmental significance of phosphorus fractions in overlying water and sediments of typical phytoplankton-and macrophyte-dominated zones. It will help to clarify the process of phosphorus migration and transformation in the sediment-water interface, and has practical significance for understanding the eutrophication process and its treatment in different regions of Taihu Lake. The investigation was conducted within typical phytoplankton-and macrophyte-dominated zones of Taihu Lake over four seasons to analyze the spatial and temporal differences between phosphorus fractions in water and sediments, and reveal their environmental significance. The results showed that:① Total phosphorus (TP), total soluble phosphorus (DTP), dissolved inorganic phosphorus (DIP), and particulate phosphorus (PP) in the overlying water of phytoplankton-dominated zones were much higher than those in macrophyte-dominated zones. Most of them showed seasonal characteristics, which were higher in summer and autumn than in winter and spring. PP is the main component of TP, accounting for 71.8% to 89.6%. A similar distribution character was found in the content of chlorophyll (Chl-a) compared with phosphorus concentration in overlying water. ② The concentration of TP in the surface sediments of phytoplankton-dominated zones was 372.38-529.64 mg·kg-1, and that in macrophyte-dominated zones was 304.29-454.27 mg·kg-1. In surface sediments, concentrations of TP in phytoplankton-dominated zones were significantly higher than in phytoplankton-dominated zones. The highest TP concentrations appeared in winter, and the lowest in summer. These were owing to the input of exogenous pollution, and the migration and transformation of internal phosphorus between sediments and overlying water under different environmental conditions. ③ The order of the mass fraction of phosphorus in sediments was:NH4Cl-P < Fe-P < Org-P < Res-P < Al-P < Ca-P. Mobile-P=NH4Cl-P+Fe-P+Org-P, accounting for 9.10%-16.93% of TP in phytoplankton-dominated zones, and slightly higher in macrophyte-dominated zones, where it was 8.11%-13.50%. Res-P accounted for 10.06%-14.97% of TP in phytoplankton-dominated zones, and 11.02%-20.28% in macrophyte-dominated zones. The risk of internal phosphorus release in phytoplankton-dominated zones is high, which is not conducive to the fixation and burial of phosphorus. The eutrophication degree of different regions in Taihu Lake is obviously different, and different characteristics of phosphorus release and burial are showed. The phytoplankton-dominated zones deserve special attention because of their high internal phosphorus load and release potential.

Alpha-Tocopherol prevents esophageal squamous cell carcinoma by modulating PPARγ-Akt signaling pathway at the early stage of carcinogenesis.

The poor prognosis of esophageal squamous cell carcinoma (ESCC) emphasizes the urgent need to better understand the carcinogenesis and develop prevention strategies. Previous studies have highlighted the potential of using Vitamin E (tocopherols) for cancer chemoprevention, but the preventive activity of α-Tocopherol against ESCC remains to be elucidated. Our data showed that early-stage supplementation with α-Tocopherol significantly prevented esophageal carcinogenesis induced by N-nitrosomethylbenzylamine (NMBA) in ESCC rat model. In the Het-1A cell model, α-Tocopherol markedly suppressed cell proliferation, promoted cell cycle G2-phase arrest and increased apoptosis. Gene microarray and proteins array analysis indicated that Akt signaling was a potential target for α-Tocopherol. We further demonstrated that α-Tocopherol increased the expression of PPARγ and its downstream tumor suppressor PTEN. Knockdown of PPARγ activated Akt signaling transduction, whereas this process was attenuated by the presence of α-Tocopherol and PPARγ agonist Rosiglitazone. In contrast, the effect of α-Tocopherol on Akt inhibition was not observed in established tumors, neither in cancerous cell lines which constitutively expressed higher levels of PPARγ. These results were closely correlated with the ineffectiveness of α-Tocopherol in the late stage of ESCC carcinogenesis. Taken together, our study suggested that α-Tocopherol may serve as a PPARγ agonist for the chemoprevention of esophageal cancer.

Xue-fu-Zhu-Yu decoction protects rats against retinal ischemia by downregulation of HIF-1α and VEGF via inhibition of RBP2 and PKM2.

BACKGROUND: Retinal ischemia-related eye diseases result in visual dysfunction. This study investigates the protective effects and mechanisms of Xue-Fu-Zhu-Yu decoction (XFZYD) with respect to retinal ischemia. METHODS: Retinal ischemia (I) was induced in Wistar rats by a high intraocular pressure (HIOP) of 120 mmHg for 1 h, which was followed by reperfusion of the ischemic eye; the fellow untreated eye acted as a control. Electroretinogram (ERG), biochemistry and histopathology investigations were performed. RESULTS: Significant ischemic changes occurred after ischemia including decreased ERG b-wave ratios, less numerous retinal ganglion cells (RGCs), reduced inner retinal thickness, fewer choline acetyltransferase (ChAT) labeled amacrine cell bodies, increased glial fibrillary acidic protein (GFAP) immunoreactivity and increased vimentin Müller immunolabeling. These were accompanied by significant increases in the mRNA/protein concentrations of vascular endothelium growth factor, hypoxia-inducible factor-1α, pyruvate kinase M2 and retinoblastoma-binding protein 2. The ischemic changes were concentration-dependently and significantly altered when XFZYD was given for seven consecutive days before or after retina ischemia, compared to vehicle. These alterations included enhanced ERG b-wave amplitudes, more numerous RGCs, enhanced inner retinal thickness, a greater number of ChAT immunolabeled amacrine cell bodies and decreased GFAP/vimentin immunoreactivity. Furthermore, decreased mRNA levels of VEGF, HIF-1α, PKM2, and RBP2 were also found. Reduced protein concentrations of VEGF, HIF-1α, PKM2, and RBP2 were also demonstrated. Furthermore, there was an inhibition of the ischemia-associated increased ratios (target protein/ß-actin) in the protein levels of VEGF, HIF-1α, PKM2, and RBP2, which were induced by Shikonin, JIB-04 or Avastin. CONCLUSION: XFZYD would seem to protect against well-known retinal ischemic changes via a synergistic inhibition of RBP2 and PKM2, as well as down-regulation of HIF-1α and a reduction in VEGF secretion.