Study on the in vitro activity of Hehuan Yin aqueous extract against hepatitis C 2a virus

Objective:To study the anti-HCV activity and mechanism of Hehuan Yin aqueous extract.Methods:Huh7.5.1 cells were used to establish the HCV2a virus infection model.Cell survival rate(%)and Renilla Luciferase Assay Kit(%)were calculated by Celltiter-GLO Assay for evaluating CC50,EC50 and SI values.To observe the drug resistance of the virus to different concentrations of Hehuan Yin within 72 hours by detecting luciferase activity,western-blot was used to detect the protein expression levels of NS5A,NS3 and NS5B.Results:the CC50,EC50 and SI of Hehuan Yin against HCV2a were 132.50g/ml,1.90g/ml and 67.90 respectively.The EC50 after 24h,48h and 72h administration were 18g/ml,5.8g/ml and 2.3g/ml respectively.Within the range of drug concentration,the aqueous extract Hehuan Yin had inhibitory effect on the expression of NS5A and NS5B proteins in a dose-effect relationship,but had no obvious effect on the expression of NS3 protein.Conclusion:The aqueous extract of Hehuan Yin may inhibit the replication of HCV2a virus by changing the protein expression levels of NS5A and NS5B,and the virus has no tolerance to the aqueous extract of Hehuan Yin.

Engineered polymer nanoparticles incorporating L-amino acid groups as affinity reagents for fibrinogen

Synthetic polymer hydrogel nanoparticles(NPs)were developed to function as abiotic affinity reagents for fibrinogen.These NPs were made using both temperature-sensitive N-isopropyl acrylamide(NIPAm)and L-amino acid monomers.Five kinds of L-amino acids were acryloylated to obtain functional monomers:L-phenylalanine(Phe)and L-leucine(Leu)with hydrophobic side chains,L-glutamic acid(Glu)with negative charges,and L-lysine(Lys)and L-arginine(Arg)with positive charges.After incubating the NPs with fibrinogen,g-globulin,and human serum albumin(HSA)respectively,the NPs that incorporated Nacryloyl-Arg monomers(AArg@NPs)showed the strongest and most specific binding affinity to fibrinogen,when compared with g-globulin and HSA.Additionally,the fibrinogen-AArg binding model had the best docking scores,and this may be due to the interaction of positively charged AArg@NPs and the negatively charged fibrinogen D domain and the hydrophobic interaction between them.The specific adsorption of AArg@NPs to fibrinogen was also confirmed by the immunoprecipitation assay,as the AArg@NPs selectively trapped the fibrinogen from a human plasma protein mixture.AArg@NPs had a strong selectivity for,and specificity to,fibrinogen and may be developed as a potential human fibrinogen-specific affinity reagent.

Discovery and identification of quality markers of Sparganii Rhizoma based on zebrafish thrombosis model

Objective:The aim of the present study was to determine the quality marker(Q-Markers)of Sparganii Rhizoma against thrombus through an integration of investigations on its antithrombotic effect,content determination and spectrum-effect correlation analysis.Methods:Based on the concept of Q-Marker,Sparganii Rhizoma was investigated for the identification of chemical component.The pharmacological effects on arachidonic acid-induced thrombosis in zebrafish were also investigated.The material basis in ethanol extract was determined by HPLC-UV.Furthermore,the potential Q-Markers were analyzed and predicted according to the effect-chemical correlation analysis.Finally,the anti-thrombotic Q-Markers were verified through the anti-thrombotic test of monomer components.Results:The model of thrombosis zebrafish was established with larvae exposed to 100 mmol/L arachidonic acid for 1 h.Nine ingredients in Sparganii Rhizoma were identified as 5-hydroxymethylfurfural,vanillic acid,ferulic acid,p-hydroxybenzaldehyde,p-hydroxybenzoic acid,vanillin,protocatechuic acid,p-coumaric acid and isoferulic acid.According to the determination effect of zebrafish thrombosis model and HPLC content analysis results,all the other contents present positive correlation except 5-hydroxymethylfurfural,and the P values of three representative potential Q-Markers(ferulic acid,protocatechuic acid and p-coumaric acid)were 0.002,0.001 and 0.026,respectively.Conclusion:Sparganii Rhizoma showed a dose-dependent effect on the recovery of reducing cardiac red blood cell on zebrafish model.Three phenolic acids(ferulic acid,protocatechuic acid and p-coumaric acid)were proved to possess the anti-thrombotic effects which could be regarded as the potential QMarkers for quality assessment of Sparganii Rhizoma.

Corrigendum to"Discovery and identification of quality markers of Sparganii Rhizoma based on zebrafish thrombosis model"[Chin.Herb.Med.13(2021)389-395]

When this paper was first published,the authors'affiliation“(“Department of Clinical Pharmacy,The First Affliated Hospital of Shandong First Medical University,Jinan 250014,China)was incom-plete.The corrected affiliation has shown above.