Résumé
ObjectiveTo investigate the relative content changes of differential metabolites and reducing sugars during the processing process of Rehmanniae Radix Praeparata (RRP) processed with Amomi Fructus (AF) and Citri Reticulatae Pericarpium (CRP), and to lay the foundation for revealing the processing principle of this characteristic variety. MethodThe samples of the 0-54 h processing process of RRP processed with AF and CRP were taken as the research object, and their secondary metabolites were detected by ultra performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The 0.1% formic acid aqueous solution (A)-acetonitrile (B) was used as the mobile phase for gradient elution (0-1 min, 1%-3%B; 1-10 min, 3%-9%B; 10-15 min, 9%-12%B; 15-22 min, 12%-18%B; 22-31 min, 18%-24%B; 31-35 min, 24%-100%B; 35-36 min, 100%-5%B; 36-40 min, 5%-1%B; 40-45 min, 1%B), column temperature was 40 ℃, injection volume was 3 μL, flow rate was 0.3 mL·min-1. Electrospray ionization (ESI) was used to scan and collect MS data in the negative ion mode, the scanning range was m/z 50-1 250. Data analysis was carried out using PeakView 1.2 software, and the chemical composition of RRP processed with AF and CRP was identified by combining the literature information and chemical composition databases. The MS data were normalized by MarkerView 1.2, and then the multivariate statistical analysis was applied to screen the differential metabolites, and the changes of the relative contents of the differential metabolites with different processing times was analyzed, finally, correlation analysis was performed between the differential metabolites, the change of the reducing sugar content was combined to determine the most suitable processing time of RRP processed with AF and CRP. ResultA total of 121 compounds were identified from RRP processed with AF and CRP at different processing times, and 12 differential metabolites were screened out by multivariate statistical analysis, including catalpol, hesperidin, isoacteoside, acteoside, narirutin, echinacoside, isomartynoside, decaffeoylacteoside, 6-O-E-feruloylajugol, dihydroxy-7-O-neohesperidin, jionoside D, and rehmapicroside. With the prolongation of processing time, the relative contents of these 12 differential metabolites and reducing sugars changed slightly at 52-54 h. ConclusionUPLC-Q-TOF-MS can comprehensively and accurately identify the chemical constituents of RRP processed with AF and CRP at different processing times, and the suitable processing time of 52-54 h is determined according to the content changes of different metabolites and reducing sugars, which provides a basis for revealing the scientific connotation of the processing principle of this variety.
Résumé
By reviewing the relevant literature of ancient herbal works and modern codices, this paper sorted out the historical evolution and developmental venation of processing of Notoginseng Radix et Rhizoma. On this basis, the modern research of processed products of Notoginseng Radix et Rhizoma was used as the breakthrough point to analyze the literature in terms of processing technology, chemical composition changes and changes in pharmacological effects before and after processing. According to the research status of processing of Notoginseng Radix et Rhizoma, some existing problems were analyzed in this paper, such as not many ancient processing methods used in modern time, lack of standardized research on processing technology. And saponins, polysaccharides, amino acids, flavonoids and other chemical components in Notoginseng Radix et Rhizoma may change to different degrees before and after processing, which was the main reason for the difference of efficacy before and after processing. However, the current research on the pharmacological effects of Notoginseng Radix et Rhizoma mainly focuses on raw products, resulting in a lack of in-depth research on the transformation mechanism of Notoginseng Radix et Rhizoma in processing difference, and the scientific connotation of "Shengxiao Shubu" has not been clearly elaborated, which is not conducive to the standardized clinical use of drugs. Therefore, it is necessary to further analyze the material basis of Notoginseng Radix et Rhizoma and its processed products, and to explore the change rule of chemical components before and after processing and its correlation with pharmacodynamic activity, so as to clarify the processing mechanism for providing scientific basis for its standardized processing, quality control and clinical rational use.
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Objective To study inhibitory effect of serine protease activity by Ulinastatin in vitro .Methods Different chromogenic peptides were designed and synthesized.Highly sensitive fluorescence detection was performed to optimize the concentration of each serine proteases and their chromogenic substrates.Multi-point method was used for the calculation of half maximal inhibitory concentration of Ulinastatin .ResuIts Ulinastain could inhibit Polymorphonuclear leukocyte elastase ( PMNE ) and plasmin with IC50 lower than 100 U/mL.For factor Xa, and Kallikrein, the IC50 of Ulinastatin was higher than 1000U/mL.No thrombin IC50 could be calculated at the present experiments.ConcIusion Similar to Ulinastatin injection from Japan, domestic Ulinastatin shows the strongest inhibitory effects on PMNE among those serine proteases.As important references, this study gives reliable data for dose range of domestic Ulinastatin in anti-inflammation, coagulation/anti-coagulation and anti-shock therapy.
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Based on introducing digital certificate related concepts, the paper compares the similarities and differences of digital sig-nature soft and hard certificates from the aspects of implementation process, implementation condition, convenience, security, law validi-ty, maintainability, etc.Combining with the actual status of information system, it analyzes the selection of digital signature.