[Optimization of Aqueous Two-Phase Extraction of Polysaccharide from Schisandra chinensis Using Response Surface Methodology with Box-Behnken Design].

Objective: To optimize the extraction condition of polysaccharide from Schisandra chinensis. Aqueous two-phase extraction（ ATPE） method was used, based on Box-Behnken design with Response surface methodology（ BBD-RSM）. Methods: solvent volume,K2HPO4 and PEG6000 were selected as the investigation factors by the single-factor experiment, and the overall desirability（ OD）of phase volume ratio, partition coefficient and extraction rates were the reponse value. BBD-RSM was used to optimize the extraction process. . Results: The optimal parameters were as follows,the solvent volume was 5 m L,the addition amount of K2HPO4 was 1. 0 g, the addition amount of PEG6000 was 1. 8 g and centrifugation time was 9 min, which indicated that the model had a good predictability. The predicted value was 0. 950,and the deviation between observed and predicted values was 3. 94% Conclusion: The ATPE technology is easy to operate and cost-effective for the extraction of polysaccharides from Schisandra chinensis.

[Study on the mechanism of haitongpi prescription extract ointment in the treatment of knee osteoarthritis based on transcriptome].

OBJECTIVE: To explore the mechanism of Haitongpi Prescription extract in the treatment of knee osteoarthritis based on transcriptome. METHODS: Total of 12 SPF grade rats were divided into control group(group C), model group(group M), and Haitongpi prescription group(group HP). The knee osteoarthritis rat model was established using the Panicker method for group M and group HP, and group HP was intervened by local topical application of Haitongpi Prescription extract for 4 weeks. Total RNA from mouse knee cartilage was extracted and three sets of differential genes were obtained through sequencing.Differential genes were prediction and analysis through GO function and KEGG pathway enrichment analysis. RESULTS: A total of 109 differentially expressed genes were identified in Group C versus Group M, while 118 differentially expressed genes were identified in Group M versus Group HP, resulting in a total of 28 genes. GO functional enrichment analysis showed that the mechanism of HP extract in treating knee osteoarthritis mainly involved immunoglobulin mediated immune response, immunoglobulin complexes, and antigen binding; KEGG pathway enrichment analysis showed correlation with tumor necrosis factor (TNF) signaling pathway, interleukin 17(IL-17) signaling pathway, and estrogen signaling pathway. CONCLUSION: HP extract can exert therapeutic effects on knee osteoarthritis through mechanisms such as immunoglobulin mediated immune response, immunoglobulin complexes, and antigen binding, as well as signaling pathways such as TNF signaling pathway, IL-17 signaling pathway, and estrogen signaling pathway.

Mechanism of icariin for the treatment of osteoarthritis based on network pharmacology and molecular docking method.

BACKGROUND: Icarin's mechanism of action in osteoarthritis (OA) was explored using network pharmacology and the GEO database, and then further validated using molecular docking. METHODS: GEO database using network pharmacology identified differential genes in OA based on Icariin's possible targets predicted by pharmmapper database. Combining the differentially expressed genes in OA with the OA-related targets, the overlapping targets were removed. In order to determine what Icariin's core targets are for treating OA, PPI network analysis was performed using OA-related targets and possible Icariin targets. Furthermore, molecular docking was used to verify the chemical's binding to the targets. Final steps included Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of genes and genomes (KEGG) pathway enrichment analysis. Cytoscape was used to construct a network of compound-target-pathway-disease. RESULTS: Protein-protein interactions between overlapping targets revealed 151 intersection targets based on a network analysis. The top ten targets with the highest enrichment scores were SRC, MAPK1, HSP90AA1, AKT1, PTPN11, ESR1, EGFR, RhoA, JAK2, and MAPK14. KEGG enrichment analysis showed that the pathways at which Icariin intervention occurs include the OA including FOXO signaling pathway, and estrogen signaling pathway. The GO analysis result showed that various biologic processes such as proteolysis, angiogenesis, innate immune response, and positive regulation of inflammatory response were involved in treatment. Molecular docking analysis confirmed that Icariin could bind well to the targets through intermolecular forces. CONCLUSION: With its multi-targeting and multi-pathway characteristics, Icariin is a promising candidate drug for treating OA.

Immunoglobulin E Epitope Mapping and Structure-Allergenicity Relationship Analysis of Crab Allergen Scy p 9.

Filamin C is an allergen of Scylla paramamosain (Scy p 9), and six IgE linear epitopes of the allergenic predominant region had previously been validated. However, the IgE epitope and structure-allergenicity relationship of Scy p 9 are unclear. In this study, a hydrophobic bond was found to be an important factor of conformation maintaining. The critical amino acids in the six predicted conformational epitopes were mutated, and the IgE-binding capacity and surface hydrophobicity of four mutants (E216A, T270A, Y699A, and V704A) were reduced compared to Scy p 9. Ten linear epitopes were verified with synthetic peptides, among which L-AA187-205 had the strongest IgE-binding capacity. In addition, IgE epitopes were mapped in the protruding surface of the tertiary structure, which were conducive to binding with IgE and exhibited high conservation among filamin genes. Overall, these data provided a basis for IgE epitope mapping and structure-allergenicity relationship of Scy p 9.

Optimisation of infrared-assisted extraction of rutin from crude Flos Sophorae Immaturus using response surface methodology and HPLC analysis.

INTRODUCTION: Rutin, one of main constituents in Flos Sophorae Immaturus, has been proven to possess several pharmacological properties such as anti-oxidant, anti-platelet, anti-inflammatory effects and so on. However, optimisation of the extraction of rutin from Flos Sophorae Immaturus has rarely been reported. Thus, it is important to develop an effective method to obtain maximum yields of rutin from Flos Sophorae Immaturus. OBJECTIVE: To develop an infrared-assisted extraction method for maximum rutin yield from crude Flos Sophorae Immaturus using response surface methodology and HPLC analysis. METHODOLOGY: Through single factor experiments, ranges of the main variables (including methanol concentration, liquid:solid ratio, extraction time and infrared power) affecting the extraction yield of rutin were confirmed. A Box-Behnken design consisting of 24 experimental runs and five replicates at zero point was then applied and a regression model was obtained to predict the optimal extraction yield. RESULTS: The ANOVA analysis indicated that the regression equation fits very well with the actual situation. The optimal conditions were as follows: infrared power 204.90 W, liquid:solid ratio 30.00 mL/g, methanol concentration 70.00% and extraction time 4.80 min. Under optimal conditions the predicted maximum yield (125.70 mg rutin/0.5 g raw material) was consistent with the experimental value (126.32 ± 0.67 mg rutin/0.5 g raw material) (n = 3). CONCLUSION: The application of response surface methodology was reliable and feasible in the optimisation of infrared-assisted extraction of rutin from crude Flos Sophorae Immaturus.

Molecular interaction studies of acetylcholinesterase with potential acetylcholinesterase inhibitors from the root of Rhodiola crenulata using molecular docking and isothermal titration calorimetry methods.

(-)-Epicatechin gallate ((-)-ECG), 1,2,3,4,6-O-pentagalloylglucose (PGG), rhodionin, herbacetin and rhodiosin isolated from the root of Rhodiola crenulata exhibited potent, dose-dependent inhibitory effects on acetylcholinesterase (AChE) with IC50 ranged from 57.50±5.83 to 2.43±0.34µg/mL. With the aim of explaining the differences in activity of these active ingredients and clarifying how they inhibit AChE, the AChE-inhibitor interactions were further explored using molecular docking and isothermal titration calorimetry (ITC) methods in the present study. Molecular docking studies revealed that all compounds except PGG showed binding energy values ranging from -10.30 to -8.00kcal/mol while the binding energy of galantamine, a known AChE inhibitor, was -9.53kcal/mol; they inhibited the AChE by binding into the ligand pocket with the similar binding pattern to that of galantamine by interacting with Glu199 of AChE. Inhibition constant of these active ingredients had a positive correlation with binding energy. The interaction between AChE and PGG was further evaluated with the ITC method and the results indicated that the PGG-AChE interaction was relevant to AChE concentration. The results revealed a possible mechanism for the AChE inhibition activity of these bioactive ingredients, which may provide some help in lead compounds optimization in the future.