Objective To investigate whether the proliferation of human hepatoma cell can be induced by insulin and which can be inhibited by Octreotide. Methods A human hepatoma cell line BEL7402 was used in this study. Using MTT method, cell count and Flow Cytometry (FCM), the effects of insulin and/or Octreotide on the number of viable cell, its ratio, the total number of cells, cell cycle and proliferative index (PI) in phasesynchronized cells were assayed. Results The cell number, the viable cell number and its ratio in insulin (0～5 ?g/ml)treated BEL7402 cells increased, while both in basic and insulininduced ones all decreased when Octreotide 0～2 ?g/ml was used, both in dosedependent manner. 1 ?g/ml of Octreotide resulted in a reduction of PI which was increased by 5 ?g/ml of insulin. Moreover, a S cell cycle block was detectable in phasesynchronised BEL7402 cell treated with 1 ?g/ml of Octreotide. Conclusion In vitro, proliferation of human hepatoma cell can be induced by insulin, while Octreotide exerted an inhibitory effects on basic and insulininduced growth of human hepatoma cell.

Thirteen of 4-anilinoquinazoline derivatives with imine groups at position 6 of quinazoline ring were synthesized and their antitumor activities were evaluated by MTT assay and Western blotting analysis. Among these compounds, 13a-131 were reported first time. The MTT assay was carried out on three human cancer cell lines (A549, HepG2 and SMMC7721) with EGFR highly expressed. Among the tested compounds, 13i and 13j exhibited notable inhibition potency and their IC50 values on three cell lines were equivalent to or less than those of gefitinib. Compound 14, without imine group substituted, displayed excellent inhibitor potency only on A549 cell line. Compounds 14 and 13j were chosen to perform Western blotting analysis on A549. The results showed that both of the compounds could inhibit the expression level of phosphorylated EGFR remarkably. It was concluded that the inhibitor potency of compound 14 was almost equivalent to that of gefitinib and the inhibitor potency of 13j was better than that of gefitinib.

Objective:To explore the effect of different separation and purification technology on the physical and chemical proper-ties of water extract of ophiopogonin. Methods:The water extraction process of total ophiopogon saponins was optimized by an orthogo-nal test. The macroporous resin adsorption and membrane separation technology were adopted to purify the ophiopogonin water extract. The physical and chemical parameters, such as electrical conductivity, pH value, viscosity and turbidity, and the contents of total sap-onins, proteins, tannins and polysaccharides were determined. Results:The optimum water extraction technology of total saponins was as follows:6-fold amount of water was added, boiling 3 times with 90 min for each time. The electrical conductivity, pH value and vis-cosity of different purified liquid of total saponins had no significant differences, while the contents of total saponins and the three kinds of macromolecules showed significant differences. Conclusion: The content of macromolecules can be used as the reference index of purification process of total saponins water extract. Compared with macroporous resin separation, membrane separation technology is more suitable for the separation and purification of total saponins water extract.

Objective: To evaluate the cold and hot characteristics of Curcumae longae Rhizoma, Curcumae Radix and Curumae Rhizoma, and study the relationship between the content of curcumin in the three herbs and the hot and cold characteristics.Methods: MTT assay was used to investigate the effect of the three herbs on the growth of SGC-7901 in vitro.Trypan-blue staining was used to analyze the cytotoxicity, and the content of curcumin was detected by HPLC.Results: Curcumae longae Rhizoma and Curumae Rhizoma could promote the proliferation of SGC-7901 at the dose of 5 μg·ml-1 and showed hot characteristics;Curcumae Radix could inhibit the growth and proliferation of SGC-7901 within the concentration range of 5-800 μg·ml-1.Trypan-blue staining showed the three herbs had no cytotoxicity on SGC-7901.The content of curcumin in Curcumae Longae Rhizoma, Curcumae Radix and Curumae Rhizoma was 4.88%, 0.15% and 0.042%, respectively.Conclusion: The content of curcumin in the three herbs affects the hot and cold characteristics, and curcumin may be the important material basis for the hot and cold characteristics.

Objective:To optimize the extraction technology and evaluate antioxidant activity of total flavonoids from Hedyotis dif-fusa. Methods:The content of kaempferol was detected by an HPLC method,and spectrophotometry was used to detect the content of total flavonoids. Results: The optimum extraction conditions were investigated by orthogonal design with the extraction quantities of kaempferol and total flavonoids as the evaluation indices. The antioxidant activities of the extracts were detected based on the clearance of hydroxyl radical and DPPH·model. The optimum extraction conditions were as follows:10-fold amount of 80% methanol and with ultrasonic extraction for 15 min. The clearance rate of 1ml methanol extract for scavenging DPPH· and ·OH was 73.89% and 91.27%,respectively.Conclusion:The optimum technology is stable and feasible for the extraction of Hedyotis diffuse. The extract shows good in vitro antioxidant properties, which provides powerful reference for the future development of relevant antioxidant prod-ucts.

Objective:To explore the macroporous resin adsorption and the membrane separation technologies for the purification of saponins water extract of Anemarrhena asphodeloides. Methods:Ten-fold amount of water was used to extract twice for 120 min each time to extract saponins from Anemarrhena asphodeloides. The macroporous resin adsorption(HP-20,HPD-600,D101,AB-8) and the membrane separation technologies (ceramic microfiltration membranes 0.8 μm and 0.05 μm, and hollow fiber ultrafiltration mem-branes 50,10 and 6 kDa) were adopted to purify the saponins water extract liquid. The physicochemical parameters including electri-cal conductivity,viscosity and turbidity were measured,as well as the contents of total saponins,proteins and polysaccharides were de-termined. Results:The viscosity and turbidity decreased,the value of pH increased and the electrical conductivity of the saponins puri-fication liquid changed irregularly after the membrane filtration. The microfiltration membrane was more advantageous than the ultrafil-tration membrane in removing macromolecular substances. The smaller the pore diameter of microfiltration membrane, the smaller the intercepted molecular weigh,the higher the removal ratio of proteins and the higher the penetration rate of the total saponins,while the polysaccharides content was stable, which was consistent with the results of physicochemical parameters. The ceramic microfiltration membrane could obtain clearer extract,while the ultrafiltration membrane was more suitable for the enrichment of saponins when the in-tercepted molecular weight was 6 kDa. The macroporous resin HPD-600 was the best for the purification of timosaponin water extraction liquid.Conclusion:The selection of membrane for the separation and purification of different substances is particularly important. The change of physicochemical parameters and the content decrease of macromolecular substances have obvious corresponding relationship. Ultrafiltration membrane is better than microfiltration membrane for the purification of timosaponin water extract liquid.

【Objective】 To explore the effect of Zhikepipa Mixture on the treatment of COVID-19 through network pharmacology analysis and molecular docking technology. 【Methods】 First, we performed the network pharmacology method to screen active compounds and targets so as to explore the mixture’s potential mechanisms in the treatment of COVID-19. In line with ADME screening index, like oral bioavailability (OB) ≥30% or drug likeness index (DL) ≥0.18, the active compounds against COVID-19 related targets were selected to construct the 'herb-compound-target’ network. Mechanism prediction of Zhikepipa mixture in the treatment of COVID-19 was analyzed by the interaction of the target sites, the bioinformatic annotation, and the metabolic pathway. Then, we used a molecular docking model to evaluate the binding ability between active compounds and 2019-nCoV (SARS-CoV-2) 3-chymotrypsin-like cysteine protease (3CL^pro) receptor-binding domain (PBD ID 6LU7), which was involved in mediating viral replication and transcription functions. 【Results】 The'herb-compound-target’ network showed 34 key compounds and 30 disease targets after overlapping with disease targets. The network topology analysis showed that those selected compounds with higher degree would produce marked anti-inflammatory effects by regulating 30 targets like PTGS1, IL1B, IL6, IL10, CXCL8 and JUN. AGE-RAGE signaling pathway, IL-17 signaling pathway, TNF signaling pathway, PI3K-Akt signaling pathway, and MAPK signaling pathway were involved in regulating hepatitis B and diabetic complications. In addition, Folium eriobotryae and Radix stemonae played important roles in the network. The molecular docking results showed that nine compounds were identified with higher docking score rank against 2019-nCoV 3CL^pro protease, and most of them were attributed to flavonoids. 【Conclusion】 Zhikepipa Mixture could exhibit both anti-inflammatory and anti-virus actions through multi-component, multi-target, and multi-pathway.

【Objective】 To investigate the active ingredients of Zizyphi Spinosae Semen (ZSS) in relieving benzodiazepine (BDZ) dependence and its molecular mechanism based on the integrated Traditional Chinese Medicine and chemical drugs idea of "enhancing effect and reducing toxicity" via the approach of network pharmacology and the technique of molecular dynamics simulation. 【Methods】 First, literature search was undertaken to find the main components of ZSS. Then, the major effective constituents of ZSS in relieving BDZ dependence and its target of action were explored on the basis of network pharmacology and molecular docking. Finally, the relationship between core components of ZSS and key proteins was further verified through the technique of molecular dynamics simulation. 【Results】 After literature search, a total of 24 chemical components in ZSS were found to act on 731 targets. Through establishing the network of "ingredients-targets-pathways" , topology analysis was performed to obtain nine core components such as linoleic acid, palmitic acid, oleic acid, tetradecanoic acid, spinosin, oleanolic acid and jujuboside A, as well as five key targets including AR, PTGS2, PPARG, RXRA and CYP19A1. Bioinformation enrichment analysis was made to obtain critical pathways such as calcium signaling pathway, cAMP signaling pathway, IL-17 signaling pathway and TNF signaling pathway. The results of molecular docking revealed that there was a good combination between core components of ZSS and key targets, and it was mainly dominated by hydrogen bonding. Furthermore, the molecular dynamics simulation experiments indicated that the combinations between jujuboside A and RXRA, oleanolic acid and RXRA, spinosin and PPARG were stable, and these three active ingredients played an important role in improving BDZ dependence. 【Conclusion】 The active components in ZSS may exert multi-target and multi-pathway intervention effects on BDZ dependence by means of processes such as immunoregulation, anti-anxiety, and anti-insomnia.