This study was designed to explore the impact of Huanglian Jiedu Decoction (HLJDT) on macrophage inflammation reaction using the network pharmacology method. Glycolysis, sphingolipid metabolism and glutamine metabolism were also investigated for "multi-component, multi-target and multi-pathway", which supports a foundation for drug innovative research. The TCMSP database was used to screen the active components of HLJDT, the target protein predicted by PharmMapper database and the DAVID database for pathways annotation and analysis. The Cytoscape 3.2.1 software was used to construct the active componenttarget-pathway network map and GENEMANIA database for protein interaction analysis. System Dock Database Site is used in verification of molecular docking. The results showed that 84 active ingredients were screened in HLJDT with a total of 111 target targets. Fourteen pathways are affected according to 13 macrophage-related inflammatory proteins, and 8 pathways including 34 target proteins from glycolysis, sphingolipid metabolism and glutamine metabolism. Inflammation-related proteins and metabolism-related proteins can interact with each other through physical correlation, protein co-expression, etc. Berberine, baicalin and geniposide combined well with 5 important targets. Huanglian Jiedu Decoction may act on the glycolysis and sphingolipid pathways to regulate macrophage inflammatory responses.

Hemorrhagic transformation (HT) is a common complication of ischemic stroke, especially after thrombolytic therapy, which is associated with increased morbidity and mortality. Thrombolysis with tissue plasminogen activator (t-PA) increases the rate of HT by as much as 10-fold, and the mortality by about 60%. The patients who are eligible for t-PA treatment are still between 3.4% and 5.2% of all patients with acute ischemic stroke because of the narrow therapeutic time window. Due to the unknown mechanism and therapeutic target of HT, there are no effective drugs to decrease the incidence of HT. The main mechanism of HT is disruption of the blood-brain barrier (BBB) integrity and neurovascular homeostasis, involving a variety of molecular signaling pathways. In animal and clinical studies, combining therapeutic agents with t-PA, which may help to minimize BBB perturbations, reduces the incidence of HT and increases the safety of thrombolytic therapy. This article is prepared to review the mechanisms, targets and therapeutic drugs of t-PA induced HT in recent years to provide a reference to the basic research and drug development of HT.

The aim of this study was to discover the pharmacological mechanism of Compound Xueshuantong in the treatment of diabetic retinopathy using network pharmacology. TCMSP software was used to search the active ingredients of Compound Xueshuantong, and the targets of its active ingredients were obtained. The targets of diabetic retinopathy were searched by OMIM, TTD, pharmGkb, DiGSeE and GAD database. The same 37 targets were analyzed by GO and KEGG using DAVID software. The results were verified using the SystemsDock. Cytoscape 3.6.1 software was used to establish an ingredient-target-pathway network model. Network pharmacological studies suggest that Compound Xueshuantong treated diabetic retinopathy through the vascular endothelial growth factor signaling pathways, mitogen-activated protein kinase signaling pathways and Toll-like receptor signaling pathways. Compound Xueshuantong alleviated diabetic retinopathy through multi-component, multi-target, and multi-pathway. This study provides a theoretical basis for further elucidation of the pharmacological mechanism of Compound Xueshuantong in the treatment of diabetic retinopathy.

Wu-tou decoction (WTD) and Baihu-Guizhi decoction (BHGZD) as described in the Synopsis of the Golden Chamber have been used extensively for the treatment of rheumatoid arthritis (RA) with apparent therapeutic efficacy. However, characteristics of pharmacological effects and their underlying molecular mechanisms have not been fully elucidated due to a lack of appropriate scientific methodology. In the current study, we performed an integrative approach applying gene expression profiling and network analysis to examine the therapeutic effects and molecular mechanisms of WTD and BHGZD based on adjuvant-induced arthritis (AIA) animal model. Results demonstrated that both WTD and BHGZD could relieve the severity of arthritis in AIA rats, while the significant differences were observed in the changes of the withdrawal response scores and latency time of AIA rats treated with WTD and BHGZD. Mechanistically, our network pharmacology-based investigation demonstrated that the major candidate targets of WTD and BHGZD were significantly associated with several inflammation-immune regulatory pathways, such as Toll-like receptor signaling pathway, T cell receptor signaling pathway, cytokine-cytokine receptor interaction, chemokine signaling pathway, B cell receptor signaling pathway, antigen processing and presentation, Fc epsilon RI signaling pathway, natural killer cell mediated cytotoxicity, as well as leukocyte transendothelial migration. In particular, the major candidate targets of WTD were also involved in the regulation of hormone and energy metabolism, which might be imbalanced during RA progression. In conclusion, the current study revealed differences and similarities regarding the effects and network regulatory mechanisms of WTD and BHGZD. These findings may present a scientific basis for elucidation of mechanisms by which WTD and BHGZD alleviates RA.

This study was designed to investigate the therapeutic effect of novel compound Z-9-octadecenyl-2-propanesulfonamide (N15) on diabetes-associated cognitive decline (DACD). Type 2 diabetes (T2DM) mice models were established with multiple injection of low doses of streptozotocin (STZ) in mice on high fat diet (HFD). Vehicle and different concentrations of N15 (50 and 100 mg·kg^-1·d^-1) were administrated orally for 6 weeks. The step-down test, dark avoidance task and Morris water maze were conducted at the 6^th week. The level of glucose and lactic acid in hippocampus were determined and mRNA of growth associated protein-43 (GAP-43), synaptophysin (SYN), brain derived neurotrophic factor (BDNF) and neurotrophins-3 (NT-3) in hippocampus were analyzed by real time PCR. The beneficial effects of N15 on learning and memory were found in the test of step-down, dark avoidance and Morris water maze. N15 reduced the level of glucose and lactic acid in hippocampus of HFD+STZ-induced diabetic encephalopathy model mice. Additionally, the mRNA expression of GAP-43, SYN, BDNF and NT-3 in hippocampus of HFD+STZ-induced diabetic encephalopathy mice were significantly increased by N15 (P < 0.01). These results suggest that the novel compound N15 can ameliorate diabetes-associated cognitive decline and the potential mechanism may be associated with the expressions of increased synaptic-related factors and neurotrophic factor in the hippocampus of diabetesassociated cognitive decline in mice.

An analytical method was developed for determination of ginkgolic acids in Yinxing Tongzhi Dropping Pills by ultra high performance liquid chromatography-triple quadrupole mass spectrometry. The samples were purified by mix-mode anion exchange and reversed-phase SPE. A chromatographic column, Waters Cortecs T3 (50 mm×2.1 mm, 2.7 μm), was used with acetonitrile-methanol-1% acetic acid (44:44:12) as the mobile phase. The ginkgolic acids were detected by electrospray ionization mass spectrometry in negative mode with multiple reaction monitoring (MRM) mode. Ginkgolic acid C13:0, C15:1 and C17:1 possessed good linear correlation in the mass concentration range from 0.2 to 200 μg·L^-1, 2 to 200 μg·L^-1, 4 to 200 μg·L^-1, respectively, with the correlation coefficients more than 0.999. The mean recoveries at spiked levels of 50, 250 and 600 μg·kg^-1 were in the range of 70.8%-95.1%, and the RSDs were 0.7%-8.6%. The limits of quantification were 1, 10, 20 μg·kg^-1, respectively. The method could be applied to the analysis of ginkgolic acids in complex matrix samples.

Jiawei Foshou San is a new Chinese medicine compound consisting of ligustrazine, ferulic acid and fumarate. Previously Jiawei Foshou San inhibited the growth of endometriosis with unclear mechanism, especially in metastasis and invasion. In this study, network pharmacology analysis was performed to explore potential mechanism of Jiawei Foshou San on endometriosis. Jiawei Foshou San compound targets were purchase from TCMID, TCMSP and SEA database. Endometriosis targets were collected from OMIM, DisGeNET and GEO database. Networks of Jiawei Foshou San compound-compound targets and compound target-endometriosis target were established with Cytoscape 3.5.0 software. Key targets were analyzed for pathway enrichment through DAVID database. It was found that Jiawei Foshou San regulated 66 core targets (MMP2, MMP9, TIMP1, ICAM1, VEGFA, et al.) and affected 115 pathways, such as estrogen, HIF-1, TNF and GnRH signaling pathways. MMP-TIMP were uncovered as one cluster of the core targets. Furthermore, Jiawei Foshou San significantly suppressed the growth of ectopic endometrium. Meanwhile, invasion and metastasis were restrained after treating with Jiawei Foshou San through decreasing MMP-2 and MMP-9, increasing TIMP-1. In brief, these results provide a pharmacodynamic basis for the study of Jiawei Foshou San.

This study was designed to explore the mechanism of total flavonoids of Astragali Radix (TFA) in treating nephrotic syndrome through establishing the active components-targets network and protein-protein interaction (PPI) networks and analyzing the functions and pathways involved in the targets. The main active ingredients of TFA were obtained by ¹H NMR and LC-MS, TCMSP and TCMID database. PharmMapper, SEA, SIB, HOME-NCBI-GENE, GeneCards and OMIM were used to predict and screen the active components of TFA. The Cytoscape software was used to construct the active components-targets network and protein-protein interactions network. The relation between the main active ingredients and targets were validated by Systems Dock Web Site. The GO and KEGG pathways involved in the targets were analyzed by ClueGO software. The target organ distribution was assigned by the BioGPS database. The results showed that 29 active components and 50 targets of TFA were screened and predicted. The network results showed that the TFA were mainly involved in biological processes such as inflammatory reaction process, oxidative stress process, apoptosis and autophagy, and played a role in the regulation of AGE-RAGE, PI3K/Akt, VEGF, IL-17 and MAPK signaling pathways to treat the nephrotic syndrome. This study reflects the characteristics of multi-components, multi-targets and multi-pathways of TFA, which provides new ideas and clues for further research on the mechanism of anti-nephrotic syndrome effects of TFA.

This study was designed to explore the anti heart failure mechanisms of the compatibility of Gualou with Xiebai based on network pharmacology in rat model of myocardial ischemia-reperfusion injury. Using the databases of Traditional Chinese Medicine Database@Taiwan (TCM Database@Taiwan), Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Drug Repositioning and Adverse Drug Reaction Chemical-Protein Interactome (DRAR-CPI) and Universal Protein Resource (Uniprot) to screen compounds and predict the target of active components, the Database for Annotation, Visualization, and Integrated Discovery (DAVID) database, we predicted the biological pathway and signal pathway in the compatibility of Gualou with Xiebai. The effects of Gualou Xiebai dropping pills on the apoptosis of myocardial cells and the expression of protein kinase B (Akt), p-Akt and cysteine aspartate-specific proteinase (caspase-3) protein were examined in the related signal pathway phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) of myocardial ischemia reperfusion injury in rats. Twenty two compounds, such as 10 α-cucurbita-5, 24-diene-3β-ol and macrostemonoside were found to protect rats from heart failure through multiple targets, multiple biological pathways and multiple pathways, involving biological pathways such as hormone stimulation reaction, phosphorylation, apoptosis regulation, and signaling pathways such as insulin, mitogen-activated protein kinase (MAPK), cell apoptosis and so on. After the intervention of Gualou Xiebai dropping pills, the PI3K-Akt signaling pathway was activated to promote the phosphorylation of Akt protein, reduce the expression of caspase-3 protein, inhibit apoptosis and protect the myocardium. The data verify the results of the network pharmacology, and explain the mechanisms of anti-heart failure activity of combination of Gualou with Xiebai.

The objective of this study is to develop an in vitro screening method for nasal absorption of insulin. First, the adaptability of in situ rat nasal perfusion test for the study of insulin was investigated. It was found that insulin was liable to be absorbed on the silicone tube and the traditional method is not suitable. However, addition of 0.001% Labrasol into the perfusate can effectively solve this problem. A modified method suitable for in situ rat nasal perfusion of insulin was established with the addition of 0.001% Labrasol into the perfusate. Using the modified method, effect of pH and drug concentration on the absorption of insulin in the nasal cavity was further investigated. The results suggest that compared with pH 4.5 and pH 7.4, the drug absorption rate was the lowest at pH 6.0. The intranasal absorption mechanism of insulin may be passive diffusion.