To explore the protective effect of protropine in Corydalis humosa Migo. on lipopolysaccharide-induced acute kidney injury in mice (AKI), an approach that used ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q/TOF-MS) coupled with a multivariate analytical platform was established. The BALB/c mice were divided into normal group (CON), model group (LPS), and protropine group (PRO). Mice were injected intraperitoneally with lipopolysaccharide solution to replicate the AKI model. Three hours after modeling, mice were given the protropine solution by gavage. Protropine was a monomer compound isolated in the laboratory, and protropine solution was prepared by dissolving protropine in sterilized distilled water. Administration was performed twice a day for three days. After modeling and administration, serum samples were collected. UHPLC-Q/TOF-MS was used to generate metabolomics data. Multivariate statistical analysis and online databases were used to screen potential biomarkers and enrich metabolic pathways. The heatmap of relative quantitative biomarker data was generated through Mev software. Animal experiments were approved by the Animal Experimentation Ethics Committee of Henan University of Chinese Medicine (No. SYXK2015-0005). The results show that the metabolic profile of mice in the LPS group was significantly altered by intervention with protropine, and clustered towards the CON group. 70 biomarkers were identified from the CON group vs LPS group (35 in positive source mode, 35 in negative source mode), and 67 biomarkers were identified from the LPS group vs PRO group (37 in positive source mode, 30 in negative source mode). A total of 34 common markers (18 in positive source mode, 16 in negative source mode) were obtained from the two comparison groups. The enrichment of all biomarkers resulted in 8 metabolic pathways including linoleic acid metabolism, D-glutamine and D-glutamate metabolism, arginine and proline metabolism, and arachidonic acid metabolism. The results show that protropine in Corydalis rhizoma ameliorates the kidney damage, insufficient energy supply, and inflammation in AKI mice by regulating amino acid metabolism, energy metabolism, and lipid metabolism in AKI mice.

Baihu-Guizhi Decoction (BHGZD), a prescription from ''Synopsis of the Golden Chamber'', has a definite clinical effect in the treatment of rheumatoid arthritis (RA). However, the research on the mechanism of this prescription mainly focuses on the regulation of inflammatory response and immune function, and its efficacy and mechanism of inhibiting synovial angiogenesis have not been reported. In the current study, transcriptomics data mining, biological network analysis and ''in vivo-in vitro'' experimental verification integrated research strategy to explore the potential and molecular mechanism of BHGZD in RA synovial angiogenesis with hot syndrome. Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of China Academy of Chinese Medical Sciences. The results of network analysis showed that the candidate network targets of BHGZD intervention in RA with hot syndrome were significantly involved in multiple angiogenesis regulation related pathways. Among them, vascular endothelial growth factor A-vascular endothelial growth factor receptor 2 (VEGFA-VEGFR2) signaling pathway contains multiple BHGZD candidate network targets, such as VEGF, phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), etc. Further experimental results showed that BHGZD could effectively reduce the expression of CD31 in knee synovium, the expression level of VEGF in serum, the activity of endothelial nitric oxide synthase (eNOS), phosphorylated VEGFR2 (p-VEGFR2), p-PI3K and p-AKT in joint tissue of adjuvant-induced arthritis rats with hot syndromes, the migration and invasion activity of HUVEC and MH7A cells, and the lumen formation activity of HUVEC cells and improve the expression level of endostatin in serum. In conclusion, BHGZD has the potential to alleviate excessive synovial angiogenesis in RA with hot syndrome, and its mechanism may be related to the intervention of VEGF/VEGFR2/PI3K/AKT signaling pathway.

This study aims to explore the mechanism of Arisaema cum Bile on epilepsy using combination of network pharmacology and patch clamp recording. Active ingredients of Arisaema cum Bile were collected from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and literatures. Epilepsy-related targets were identified from GeneCards and OMIM database. STRING platform was employed to perform protein-protein interaction (PPI), and David platform was used for Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) enrichment analysis. The drug-ingredient-target-pathway-disease network was constructed with Cytoscape software. The animal experiments were approved by the Laboratory Animal Ethical and Welfare Committee Hebei Medical University. Nine compounds were detected as the active ingredients of Arisaema cum Bile, including chenodeoxycholic acid (CDCA), deoxycholic acid and β-sitosterol, etc. A total of 22 key potential targets were identified, including 5-hydroxytryptamine transporter (SLC6A4), gamma-aminobutyric acid receptor type A subunit alpha 2 (GABRA2) and neuronal acetylcholine receptor subunit alpha-7 (CHRNA7). These targets were associated with biological processes of serotonergic synapse, GABAergic synapse and ion transmembrane transport. Brain slice electrophysiology experiments revealed that β-sitosterol and CDCA inhibited the action potential (AP) of CA1 pyramidal neurons in the mouse hippocampus. Both β-sitosterol and CDCA affected the properties of AP, such as rheobase, delay and depolarization duration. In addition, the inhibitory effect of AP was more prominent when the two compounds were given together. Combining with network pharmacology and electrophysiological experiments, our study reveals the potential mechanisms of Arisaema cum Bile for the treatment of epilepsy in a "multi-ingredients, multi-targets and multi-pathways" manner. Our study provides a reference for further studies of Arisaema cum Bile.

To study the multivariate statistical analysis and metabolic regulation mechanism of rhubarb in the treatment of blood stasis syndrome through the ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF/MS). In this study, multivariate statistical analysis was used to comprehensively analyze the differential characteristics of metabolites with significant regulatory effect in plasma samples of blood stasis model rats, analyze the changes of differential metabolites of rhubarb regulating blood stasis syndrome, and analyze the pathway enrichment of identified metabolites. The results showed that the differentiation of plasma differential metabolites between the model group and the control group was good (Q² > 0.5), and 44 differential metabolites all had different degrees of callback. The expressions of PC(15∶0/20∶2(11Z, 14Z)), PC(18∶1(9Z)/18∶1(9Z)), salicylaldehyde and glycolic acid were down-regulated in the rat model of blood stasis syndrome, and rhubarb could up-regulate them. The expression levels of (±)8-HETE, taurodeoxycholic acid and γ-murocholic acid were up-regulated in the blood stasis rat model, which could be down-regulated by rhubarb. Differential metabolites are enriched in 97 metabolic pathways, involving lipid metabolism pathways, inflammatory factors and immune pathways, and steroid hormone synthesis pathways. This study clarified the mechanism of rhubarb in the treatment of blood stasis syndrome from the perspective of plasma metabolism, and provided a theoretical basis for the further development and clinical application of rhubarb. This study has been approved by the experimental animal Ethics Committee of Shaanxi University of Traditional Chinese Medicine (No. SUCMDL202103009002).

Invasive fungal infections are a tricky problem with high morbidity and mortality. The emerging of drug resistance worsens this problem. Therefore, developing novel antifungal agents is an urgent need to break these hurdles. Although protein kinase inhibitors have been extensively investigated in various fields such as cancer, diabetes, rheumatosis and so on, they are less explored in fungal infection. Previously, we found 12 protein kinase inhibitors with different antifungal activities, among them 5-iodotubercidin (5-Itu) was found to be the most potent antifungal agent with minimal inhibitory concentrations ranging from 2 to 4 μg·mL^-1. Moreover, 5-Itu displayed potent fungicidal effect, inhibition of hyphal formation and anti-biofilm activity. The mechanistic studies indicated that 5-Itu changed membrane sterol compositions and ultrastructures, increased the cell membrane permeability and induced apoptosis. Therefore, it needs to further study 5-Itu for the discovery of promising antifungal lead compounds.

Totally 28 new 12N-substituted aloperine derivatives were designed, synthesized and evaluated for their down-regulating PD-L1 activities in breast cancer MDA-MB-231 cells. Among them, compound 7f could significantly down-regulate PD-L1 level in concentration- and time-dependent manners, and exhibit a low cytotoxicity. It activated the killing activity of co-cultured T cells against tumor cells in a concentration-dependent manner, showing the potential of tumor immunotherapy. Further study indicated that 7f mediated the degradation of PD-L1 through a lysosomal pathway. This study provides useful guidance for the development of aloperine compounds into new small molecule tumor immune suppressants.

Electrospray ionization (ESI) is easy to be affected by the biological matrix interferences, and thus the accuracy, precision, and reproducibility of the quantitation are significantly impaired. Probe electrospray ionization (PESI) is one of the most typical ambient ionization, which can ionize molecules without complicated sample preparation at the atmospheric environment, and is superior in simplicity, high efficiency, and high throughput. The micro pen electrospray ionization tandem mass spectrometry (μPen-ESI-MS/MS) method was newly developed based on PESI. In this study, the matrix effect of the μPen-ESI-MS/MS method for drug quantitation in plasma samples was evaluated and compared with the liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) method. The μPen-ESI-MS/MS and LC-ESI-MS/MS methods for quantitation of five drugs in rat plasma were established respectively, and the matrix factors (MFs) and internal standard normalised matrix factors (IS normalised MFs) were measured. The results showed that the ion suppression of the μPen-ESI-MS/MS method for tacrolimus, flunarizine, and desloratadine was equal to or less than that of the LC-ESI-MS/MS method; the RSDs of the IS normalised MFs of all five drugs were less than 15%, which met the related requirements of the Pharmacopoeia of the People's Republic of China. Therefore, this study investigated the matrix effect of the μPen-ESI-MS/MS method for quantitative analysis of target drugs in plasma samples and provided new data for the application of μPen-ESI-MS/MS in bioanalysis.

To investigate the effect of ionic liquid based on choline and citronellic acid ([Cho] [CA], COCA) on the oral absorption of poorly soluble drug cyclosporin A (CsA), COCA was synthesized using choline and citronellic acid by one-step neutralization method, and then characterized by mass spectrometry, ¹H-NMR, and infrared spectrophotometry. Next, CsA-ionic liquid (CsA-COCA) was prepared by ultrasonic-assisted method and filled into enteric-coated capsules. After oral administration of CsA-COCA capsules or CsA suspension preparation in rats, CsA concentration in whole blood was assayed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, and the pharmacokinetic parameters were calculated by DAS 2.0 software. All animal care and experiments followed the approval of Institutional Animal Care and Use Committee at Peking University Health Science Center. The results indicated that compared with oral administration of 10 mg·kg^-1 CsA suspension, the area under the curve (AUC), half-life time (T_1/2), and mean residence time (MRT) of CsA obtained by oral administration the same dose of CsA-COCA increased by 2.81, 4.41, and 1.77 times, respectively. The COCA prepared in this study can significantly promote the oral absorption of CsA in rats, and prolong the half-life. This study can provide reference for the study of oral formulation of insoluble drugs such as CsA.

Multicellular tumor spheroids (MCTS) can simulate the structure and metabolic characteristics of tumors in vivo, which is of great significance to study the metabolic phenotype of tumor cells and the mechanism of drug intervention. In this study, esophageal cancer MCTS were constructed, and MCTS frozen sections were prepared after treated with different formulations of paclitaxel (PTX) including common PTX injection, PTX liposome and albumin bound PTX. MCTS mass spectrometry imaging analysis method was established by using air flow assisted desorption electrospray ionization mass spectrometry imaging (AFADESI-MSI). The visualization of the permeation and enrichment process of PTX in MCTs after PTX treatment was realized, and the spatially resolved metabolomics of PTX injection group was studied. The results showed that the permeation and enrichment behavior of PTX in MCTs model were related to the formulations. The changes of endogenous metabolites in MCTs of esophageal cancer after treated with PTX injection had temporal and spatial characteristics. The metabolic changes of MCTS during the initial 0-4 hours were dominated by the down-regulation of middle-high polarity metabolites and some lipids in the central region of MCTS, while the metabolic changes of MCTS during 8-72 hours were mainly up-regulated by lipid metabolites in the peripheral region of MCTS. The combination of in vivo tumor-associated MCTs model with label free, highly sensitive and high coverage mass spectrometry imaging technology provided a new method and strategy for the study of pharmacometabolomics.

Aa a characteristic medicinal plant in China, Gentiana rigescens Franch. has the function of protecting the liver and invigorating the spleen. At present, there are a few studies on the content determination method of characteristic components of G. rigescens, so it is necessary to establish a scientific and effective quality control method; In this study, The high performance liquid chromatography (HPLC) fingerprint of G. rigescens was established, based on literature reviewed and characteristic spectrum identified, the source range of G. rigescens quality marker (Q-marker) was screened. The effectiveness of the ingredients and the corresponding targets and pathways was analyzed through network pharmacology, and drew the diagram of ''component-target-pathway''. Qualitative and quantitative analysis of G. rigescens was performed by HPLC, and screen the main marker components leading to the differences between groups which were determined the Q-marker of G. rigescens; The literature and HPLC had determined that five iridoids were the main source of G. rigescens Q-marker. The network pharmacology (effectiveness) and qualitative and quantitative (detectability) analysis of G. rigescens from different producing areas confirmed that gentiopicroside, swertiamarin, and sweroside can be used as the main landmark components, and there were significant differences in their contents among different producing areas; The analysis of G. rigescens from different producing areas was carried out by network pharmacology and chemical fingerprints, it is confirmed can be used as potential Q-marker to provide sufficient theoretical basis for the quality control of G. rigescens in the later period.