The molecular mechanism of spleen-strengthening and moisture-nourishing liver prescription in treatment of acute-on-chronic liver failure based on network pharmacology and experimental verification / 中国药理学通报

To explore the mechanism of spleen- were obtained for the treatment of acute-on-chronic livstrengthening and moisture-nourishing liver prescription er failure, and 244 intersecting target genes and 7 core (JPLSYGF) in the treatment of acute-on-chronic liver target genes were screened. Molecular docking showed failure using network pharmacology and the molecular that the core target genes AKT1, SRC, VEGFA, docking. Methods Relying on TCMSP and Gene- STAT3 , EGFR, MAPK3 , HRAS had good affinity with Cards and other databases, the relevant targets of JPL- quercetin, the main active component in the JPLSYGF in the treatment of acute-on-chronic liver failure SYGF, and had strong binding activity. In addition, in were obtained. String and Cytoscape were used to con- vivo tests verified that the JPLSYGF could reduce the struct PPI networks of targets, core targets were expression of HRAS, EGFR, STAT3 , SRC, and VEGscreened out, and DAVID was used for GO function FA, to delay the progression of acute-on-chronic liver annotation and KEGG pathway enrichment analysis. failure. Conclusions JPLSYGF may act on core tar- The main active ingredients of the traditional Chinese gets such as HRAS, EGFR, STAT3, SRC, VEGFA medicine compound formula for JPLSYGF were select- and so on, to achieve the effect of treating acute-oned with a bioavailability OB value of =Э 30% and a chronic liver failure. drug-like DL

The molecular mechanism of Qizhu anti-cancer prescription in treatment of primary liver cancer based on network pharmacology and experimental verification / 中国药理学通报

Aim To investigate the mechanism of Qizhu anti-cancer prescription ( QZACP) inthe treatment of primary liver cancer using network pharmacology and molecular docking. Methods Drugs and primary liver cancer ( PLC) -related targets were found according to TCMSP database and disease databases such as GeneCard, the key chemical components and core targets were screened by Cytoscape 3. 9. 1 and String platform respectively, and a network relationship diagram of traditional Chinese medicine-active component-target was constructed by using Cytoscape 3.9. 1. GO functional analysis and KEGG pathway analysis were performed using DAVID platform, visualized by R 4. 1. 1 software, and finally the core clustered proteins were analyzed by CytoNCA plug-in to obtain the core action targets, and the core components and key targets were verified by using molecular docking technology and the pharmacodynamic mechanism of QZACP was further verified by animal experiments. Results The active ingredients of QZACP in the treatment of primary liver cancer may be quercetin, glycyrrhizin, Denudatin B, isoflavanone, sanguinarol, etc. ; the potential targets were STAT3, EGFR, AKT1 etc. ; the related pathways were mainly PI3K-Akt signaling pathway,MAPK signaling pathway,etc. ; molecular docking showed that the core compounds had better integrating conformation with the key targets. In addition, QZACP could inhibit the growth of tumor in nude mice and decrease the expression of STAT3, EGFR and AKT1. Conclusions Qizhu anti-cancer prescription may have some positive significance in the treatment of primary liver cancer, which may be related to the regulation of PI3K/Akt signaling pathway.

Rapid identification of classical herbal formulae Baoyinjian's chemical substance based by HPLC-Q/TOF-MS / 中草药

Objective: To analyze the chemical constituent cluster of classical herbal formulae Baoyinjian systemically by HPLC-Q/TOF-MS. Methods: The seperation was performed on Diamonsil C18 (250 mm × 4.6 mm, 5 μm) column with gradient elution with acetonitrile-0.1% formic acid. The column temperature was 30 ℃, the flow rate was 1 mL/min, the injection volume was 10 μL, and the mass spectrometry condition was X500R QTOF mass spectrometer, electrospray ion source, positive and negative mode scanning. Results: A total of 52 chemical constituents were identified by reference confirmation, literature comparison, and high mass spectrometry data analysis. The chemical constituent cluster was composed of 17 flavonoids, six phenolics, 12 iridoid glycosides, eight alkaloids, one phenethyl alcohol glycosides, four monoterpene glycoside, two triterpenes and two other compound. Conclusion: This study can identify various chemical constituents of Baoyinjian systematically, accurately, and rapidly, which provides a basis for the determination of the quality attributes of Baoyinjian.

Practice and progress on bionic technology in identification on five tastes of Chinese materia medica / 中草药

Four properties and five tastes are the core content of medicinal theory of Chinese materia medica (CMM), and it is important for modern study of CMM to elucidate the scientific connotation of five tastes based on the application of bionic technology. With the development of bionic technology, electronic nose and electronic tongue have become the objective method to determine the taste and smell in recent years. Based on the taste and odour connotation of five tastes of CMM, the working principle, research method, and identification of flavor substances in CMM of electronic nose and electronic tongue were summarized, and the progress on original variety, production place, growth period, storage life and processing technology of CMM were reviewed in this article. Combined with the practice of our research group on the characterization of medicinal properties, the basic research model of the definition and characterization of the five tastes of effective materials basis was proposed, which would provide a reference method for the medicinal research on five tastes of CMM.

Koumine-induced apoptosis of human colonic adenocarcinoma cells: the cell biological mechanism / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the cell biological mechanism of koumine-induced apoptosis of human colonic adenocarcinoma cells.</p><p><b>METHODS</b>The effects of LoVo cell koumine on the membrane potential, mitochondrial membrane potential, concentration of cytosolic free calcium, reactive oxygen species (ROS) and gap junctional intercellular communication was observed by laser scanning confocal microscopy.</p><p><b>RESULTS AND CONCLUSION</b>Koumine lowered the membrane potential and mitochondrial membrane potential of LoVo cells and decreased the concentration of free cytosolic calcium, which was increased to a level higher than the basal one after addition of EDTA. Koumine also increased the reactive oxygen species and enhanced the gap junctional intercellular communication of LoVo cells. These findings may explain the possible mechanisms of koumine-induced LoVo cell apoptosis.</p>

Over-expression in Escherichia coli and characterization of apolipoprotein AI / 生物工程学报

Apolipoprotein AI (apo AI), the major protein component of human high-density lipoprotein (HDL), is a single-chain polypeptide of 243 amino acids. Several epidemiological studies have shown that the plasma concentrations of HDL has the role of reverse cholesterol transport (RCT) and inversely correlated with the incidence of coronary artery disease. Because apo AI lacks post-translational modifications, it is convenient to express human apo AI in Escherichia coli expression system. However, there is a poor stability of the mRNA and the apo AI protein in E. coli, it is difficult to express mature apo AI in recombinant bacteria, moreover, even as a fusion protein, apo AI is still sensitive to degradation and can not be cleaved efficiently from the fusion tags. In contrast, proapolipoprotein AI (proapo AI, having an additional polypeptide containing the amino acids Arg-His-Phe-Trp-Gln-Gln at the amino-teminal of the mature protein) proved stable and undegraded in Escherichia coli, and therefore, in this research, an expression system of E. coli including a plasmid of P(R)P(L) tandem promoter was adapted to produce proapo AI. Furthermore, site-directed mutagenesis of the proapo AI cDNA was performed to generate a Clu8Asp mutation in the amino-terminal sequence of proapo AI which created an acid labile Asp-Pro peptide bond between amino acid 8 and 9, and permitted specific chemical cleavage to remove pro-peptide. After inducing with a shift of temperature, yields of recombinant proapo AI achieved about 40% of total cell protein and the recombinant proapo AI expressed proved as a form of inclusion body in cells, so protein need to renature. First of all, the protein was dissolved in buffer with denaturant, and renaturation was carried out on a hydrophobic interaction column (Phenyl Sepharose), ion-exchange chromatography and gel-filtration chromatography were then used to further purify the protein. The purified recombinant apo AI was detected by a set of tests including Western-blotting, Circular dichroism spectra and lipid-binding test, the results shown that recombinant apo AI has similar structural and lipid-binding properties identical to those of native plasma apo AI, which facilitates further research and application.

Study on the Recombinant Human ApoA-I_Milano High Density and High Expression by Two Temperature-Shifted Induction in Escherichia coli / 微生物学通报

The temperature effect on the recombinant protein production formation was investigated in present study. The culture temperature of growth phase is 30℃, and the culture temperature of induction phase was arranged according to three modes. Hign cell-density and high expression culture of E.coli to product recombinant human apolipoprotein A-I Milano by two temperature-shifted induction . Two temperature-shifted induction was carried out high density and high expression recombinant human ApoA-1 Milano. The recombinant protein ApoA-I Milano reached 4.8 g?L -1 with the final cell density of OD 600 150. And the two temperature-shifted induction avoided the acetic acid successfully to the influence of the high density and high expression. Two temperature-shifted induction was viable in high density culture and high expression of heterogenous protein in recombination E.coli.The sduty provides a basic work for production of recombinant ApoA-I Milano in scale.

Optimation of the Fermentation Conditions for Human ApolipoproteinA-I Expression / 微生物学通报

Optimization of the fermentation condition for human apolipoproteinA-I expression in recombinant Escherichia coli was investigated. The recombinant plasmid pBV220-ApoA-I was transformed respectively into different E.coli hosts such as JM109, BL21(DE3),DH5?, BMH7118,and TG1. The best host E.coli was DH5? in which the recombinant ApoA-I expression percentage was 21.2% corresponding to that in BL21(DE3) in flask shaker cultivation,while the ApoA-I expressed percentage in E.coli TG1 was 11%.Fed-batch cultivation was performed in FMG-5L fermentor,the optimum fermentation cultivation conditions were as following :optimum pH value was 7.0 in growth phase and 7.4 in the expression phase. The initial glucose concentration in batch phase was 3 g?L -1.The optimum C/N ratio was 2∶1.The recombinant ApoA-I reached about 40% of the total protein, and concentration of ApoA-I was 2.86 g?L -1.