OBJECTIVE: Salt-processed Psoraleae Fructus (SPF) is widely used as a phytoestrogen-like agent in the treatment of osteoporosis. However, due to improper clinical use or misuse, resulting in liver damage. In this study, network pharmacology was employed to analyze the mechanism of cholestatic liver damage. An adeno-associated virus overexpressing SULT1E1 (rAAV8-SULT1E1) was constructed and the hepatotoxicity of SPF, psoralen, and isopsoralen was determined. METHODS: By utilizing three databases inclding TCMSP, TCMID, and BATMAN- TCM, the targets of the three databases were summarized, and a total of 45 psoralen compounds were included. Network pharmacology analysis was then performed. The adenoviral vectors were injected into the tail vein of C57BL6 mice to elucidate the role of SULT1E1 in SPF-induced cholestasis-mediated hepatotoxicity in vivo. SPF (10 g/kg), psoralen, and isopsoralen (50 mg/kg each) were intragastrically administered to mice for 30 d. B-ultrasound and samples were collected and examined for follow-up experiments. RESULTS: A total of 854 targets were predicted for 45 active components, with 151 cholestasis-mediated hepatotoxicity-related disease targets obtained for SPF. A total of 126 pathways were enriched based on KEGG pathway analysis, with the "estrogen signaling pathway" identified as one of the top 20 pathways. In terms of pathological hepatic changes, treated mice had visually swollen hepatocytes, dilated bile ducts, and elevated serum biochemical markers, which were more prominent in mice treated with isopsoralen than in those treated with other compounds. Notably, the overexpression of SULT1E1 could reverse liver damage in each treatment group. B-ultrasound was used to observe the size of the gallbladder in vivo. The size of the gallbladder was found to significantly increase on day 30 after treatment in the SPF-, psoralen-, and isopsoralen-treated groups, especially the SPF group. Compared with the expression levels in the negative control group (rAAV8-empty + con), the expression levels of FXR, Mrp2, Bsep, SULT1E1, SULT2A1, Ntcp, and Nrf2 decreased, whereas those of CYP7a1 and IL-6 increased in the SPF-, psoralen-, and isopsoralen-treated groups. CONCLUSION The overexpression of SULT1E1 could alleviate the decreased or increased expression of indicators, indicating that SULT1E1 is an important target gene for SPF-induced liver damage. The severity of liver damage was significantly lower in the rAAV8-SULT1E1 groups than in the rAAV8-empty groups.

Objective To express the protein enconded by the Rv3432c gene of Mycobacterium tuberculosis(M.tb)in vitro by prokaryotic expression,to analyze the structure of the Rv3432c protein by using bioinformatics software,and to explore for new drug targets against M.tb.Methods The Rv3432c gene was amplified by PCR using the genomic DNA of the inactivated M.tb strain H37Rv as the template and a recombinant plasmid was constructed with the expression vector pET-28a.The expression products were analyzed by SDS-PAGE and purified using affinity chromatography.The biological properties of Rv3432c were analyzed with Protparam,the Pfam online tool,SOMPA,Protscale,TMHMM Signalp 6.0,NetPhos3.1,SUMOsp 2.0,and SWISS-MODEL.Results pET-28a-Rv3432c recombinant plasmid sequencing results were fully consistent with those of the target gene.SDS-PAGE analysis showed that the fusion protein existed in the form of a soluble protein with a relative molecular mass of about 55×103,which matched the expected size.ProtParam analysis showed that the Rv3432c protein was hydrophilic(showing a GRAVY value of-0.079).Rv3432c was a protein with no transmembrane structural domains or signal peptide.The secondary structure of Rv3432c mainly consisted of random coils(39.78%)and α-helix(39.57%)and was relatively loosely structured.Conclusion We successfully constructed a prokaryotic expression plasmid of the Rv3432c protein and analyzed its structure using bioinformatics,laying the foundation for further research on the role of Rv3432c in the pathogenesis and progression of tuberculosis as well as the identification of new drug targets against M.tb.

In order to improve the poor solubility and low bioavailability of paeonol (Pae), paeonol-nanoemulsion (Pae-NE) was prepared, and its effect on uptake of human umbilical vein endothelial cells (HUVECs) was investigated.Pae-NE was prepared by phase inversion composition (PIC), the formulation of Pae-NE was optimized by single factor method and central composite design-response surface method (CCD), and the pharmaceutical properties were further characterized.Moreover, MTT was applied to evaluate the toxicity of Pae-NE on HUVECs, and the cellular uptake efficiency of Pae-NE was detected by fluorescence microscopy and flow cytometry.The results showed that the optimal formulation of Pae-NE was 20 mg of Pae, 55.1 mg of LCT, 144.9 mg of MCT, 600 mg of HS15, and 200 mg of 1,2 propylene glycol.The Pae-NE appearance was a light blue emulsion, and the average particle size is (25.69 ± 0.03) nm, with PDI of 0.182 ± 0.09, Zeta potential of -(4.01 ± 0.30) mV and good stability.The drug loading of Pae-NE was (1.967 ± 0.28) mg/mL and encapsulation rate of (99.36 ± 0.1)%.Pae-NE performed no significant effect on HUVECs growth in the Pae concentration range of 10-1-10-3 μg/mL.Moreover, NE as a drug delivery carrier significantly enhanced the uptake efficiency of Pae on HUVECs.In conclusion, Pae-NE preparation method was simple and stable, and promotes HUVECs uptake efficiency of Pae, suggesting that NE was a better dosage form reference for the lipid-soluble drug of Pae.

China ; Humans ; Lenalidomide/therapeutic use* ; Multiple Myeloma/drug therapy* ; Thalidomide ; Therapeutic Equivalency

Objective:To establish an HPLC method for the determination of content and entrapment efficiency of HIV-1 virus in-fection factor Vif inhibitor VEC-5 liposomes. Methods:VEC-5 liposomes were prepared by a method of freeze-drying and reconstruc-tion. The separation of free drug from the liposomes was achieved by ultracentrifugation, and an HPLC method was used to determine the content and entrapment efficiency of VEC-5 liposomes. Results:The linear range of VEC-5 was 20-100 μg·ml-1(r=0. 999 0). The average recovery was 100. 25% and RSD was 0. 93%(n=9). The content of three batches of VEC-5 liposomes was 98. 63%, 100. 43% and 102. 65%, respectively within the range of 90%-110%, and the entrapment efficiency was 94. 89%, 93. 68% and 94. 56%, respectively, which was above 90%. Conclusion:The method is accurate and reliable, which can be used to determine the content and entrapment efficiency of VEC-5 liposomes.

Objective: To find out a new clarification process of Kechuanning Oral Liquid by chitosan, and to compare with the process with ethanol sediment. Methods: Glycyrrhizinate, papaverine hydrochlorid were qualitatively analysed and ephedrine hydrochloride was quantitatively determined in the two processes. Stability of the two preparations were compared. Results: Both the processes by chitosan and by ethanol have clearing action the former reserved more effective components than the latter. Conclusion: the process by chitosan can substitute for process by ethanol sediment in production of Kechuaming Oral Liquid.