This study was aimed to improve the drug activity of three kinds of isoflavones from Chickpeas.Biochanin A,formononetin and genistein were used as raw materials.Acetone was used as solvent.Potassium carbonate was used as catalyst.The etherification reaction was with 1,3-dibromopropane,1-bromopropane and 3-bromopropene.The results showed that 9 isoflavone ramifications were synthesized.This method was simple,easy to control with high yield.It was concluded that the product structure was confirmed by 1H-NMR,13C-NMR and ESI-MS analysis.It laid a foundation for the structural study basis in the further research of its drug activity.

Objective To establish the HPLC fingerprints of Tangbikang Granules; To scientifically evaluate and effectively control the quality of Tangbikang Granules; To ensure its production stability. Methods HPLC was performed on the column of Germany Merck RP-18 endcapped (250 mm × 4.6 mm, 5 μm) with mobile phase of acetonitrile-0.1% formic acid water; column temperature was 40 ℃; flow rate was 1.0 mL/min; detection wavelength was 240 nm; volume injection was 20 μL. Fingerprint Similarity Evaluation Software (edition 2004A) of Chinese Pharmacopoeia Commission was used to evaluate the similarity of the 10 batches of Tangbikang Granules, and to analyze the correlations of 9 ingredients in Tangbikang Granules. Results Wogonoside was used as the reference peak, and the common mode for the HPLC fingerprints was set up. The similarities of the 10 batches of Tangbikang Granules were above 0.930, and altogether 25 common peaks in the chromatograms were found, of which 18 peaks were assigned to Chinese materia medica in Tangbikang Granules. Conclusion The method has good separability and is accurate and simple, which can provide references for the quality control of Tangbikang Granules.

This study was aimed to observe the effects of gymnema sylvestre total saponins (GA) on insulin resistance (IR) in adipose tissues of KKay mice,in order to discuss the action mechanism from the PI3K/AKT signal transduction pathway.A total of 27 KKay mice were randomly divided into the model group (DM),Pioglitazone group (BG) and GA group.Nine normal C57BL/6J mice were used as the normal control group (NC).Intragastric administration of drugs was given for eight weeks.The bodyweight and food intake of all mice were tested each week.After the experiment was completed,fasting plasma insulin (Fins) and fasting plasma glucose (FPG) were detected and the insulin sensitive index (ISI) was calculated.Expressions of PDK-1,AKT,P-AKT (Ser473),P-AKT (Thr308) in adipose tissues of epididymis in mice were detected.And expressions of PI3K-p85 mRNA,PTEN mRNA,APN mRNA were also measured.The results showed that compared with DM group,bodyweight of BG and GA groups were decreased (P＜0.05,P＜0.05);FPG and Fins level of BG and GA groups were decreased (P＜0.05,P＜0.05,P＜0.05,P＜0.05),ISI increased (P＜0.05,P＜0.05);APN mRNA increased in BG and GA group (P＜0.01,P＜0.01);PI3K-p85 mRNA increased in GA group (P＜0.05);P-AKT (Thr308) protein expression increased in BG and GA group (P＜0.05,P＜0.01);P-AKT (Ser473) protein expression increased in GA group (P＜0.05);the phosphorylation of AKT (Thr 308) was enhanced in BG and GA group (/9＜0.01,P＜ 0.01);the phosphorylation of AKT (Ser473) was enhanced in GA group (P＜0.01,P＜0.01).PDK-1 protein expression was decreased in BG and GA group (P＜0.05);PTEN mRNA decreased in BG and GA group (P＜0.05,P＜0.01).It was concluded that GA can ameliorate IR by sensitizing PI3κ/AKT signal pathway in adipose tissues of KKAy mice.

This paper was aimed to study the albiflorin activity of Tang-Bi-Kang (TBK) granules in protecting Schwann cells (SCs) of rat's sciatic nerve.The establishment of SCs oxidative stress model was the condition of 150 mmol×L-1 Dglucose with different concentrations.And the incubation time was 48 h.The experiment groups were the high-dose,middle-dose and low-dose (100 μM,20 μM,4 μM) albiflorin group,the model group,the vitamin C (100 μM) group,and the normal group.Flow cytometry was used to detect the content of ROS in SCs.Fluorescence microscope was used to observe the condition of ROS fluorescence in SCs.And CCK-8 was used to detect the cell activity.The results showed that by using CCK-8 to detect cell proliferation,after 48 h,there was a significant difference between the model group and the normal group (P＜0.01);the albiflorin group compared with the model group (P＜0.01).It indicated that albiflorin can promote the proliferation of SCs.Detecting the ROS fluorescence content,it showed that compared with the model group (Glu 150 mM),the 100 μM,20 μM,and 4 μM albiflorin group,it was P＜0.01 for each group.It showed that albiflorin could relieve the ROS in SCs and alleviate oxidative stress.It was concluded that albiflorin can increase the proliferation of SCs and improve the state of oxidative stress with the protection of SCs.

Through comprehensively characterizing components in blood after oral administration of Tang-Bi-Kang (TBK) granules by UPLC-ESI-MSn,this study was aimed to explain the pharmaceutical material basis of TBK initially.UPLC-LTQ-Orbitrap was used under both positive and negative ion modes of electrospray ionization.The blank serum and rat serum after oral administration of TBK were analyzed.Components in rat serum were identified and characterized based on ion fragment information,evenelectron law,nitrogen rule and so on.Reference data was used to establish the UPLC-ESI-MSn method.The results showed that after oral administration of TBK granules,15 components were detected in the serum,of which 13 components were taken as the prototype to blood and 2 metabolites.It was concluded that constituents of TBK granules in rat serum were generated from compatibility of all herbal medicines.In rat serum,most of the components had been absorbed by rat's metabolism;a few were absorbed as the prototype.This research provided references for pharmacodynamic material basis and metabolism of TBL granules in vivo.

This paper was aimed to study the effect of ethyl acetate extracts of Gentianella acuta on the gene and protein of insulin significant signal IRS-1 and Akt in insulin resistance (IR) HepG2 cells.The CCK-8 method was used to detect the HepG2 cell activity.HepG2 cells of human liver cancer were cultured with high concentration insulin (10-6 mol· L-1)for 36 hours to establish IR cell model.According to the results of CCK-8,the control group,model (IR) group,ethyl acetate extracts of Gentianella acuta IR + 50 μg· mL-1,IR + 500 μg· mL-1 group,and the metformin group were divided.Glucose consumption was measured with a glucose assay kit.The expressions of IRS-1 and Akt gene in IR HepG2 cells were detected by RT-PCR after 6-hour using of ethyl acetate extracts of Gentianella acuta.Western blot was used to detect the expression of IRS-1 and Akt protein after 6-hour using of ethyl acetate extracts of Gentianella acuta.The results showed that when the concentration of ethyl acetate extracts of Gentianella acuta was 500 μg· mL-1,the survival rate reached 95％.When the concentration was higher than 500 μg· mL-1,the survival rate decreased.Compared with the IR group,the IR + 50 μg· mL-1 group and the IR + 500 μg· mL-1 group promoted glucose consumption of IR HepG2 cells,but its effect was less than that of the metformin hydrochloride group.The expression of IRS-1 and Akt in IR HepG2 cells was significantly increased by using RT-PCR in the group of IR + 50 μg· mL-1 and IR + 500 μg·mL-1 compared with the IR group after 6-hour using of ethyl acetate extracts of Gentianella acuta.The expression of IRS-1 and Akt protein in the group of IR + 50 μg· mL-1 and IR + 500 μg· mL-1 was significantly higher than that in the IR group after 6-hour medication detected by western blot.It was concluded that the ethyl acetate extracts of Gentianella acuta can increase the expression of IRS-1,Akt gene,the expression of IRS-1 and Akt protein in HepG2 cells,which may be the mechanism of IR improvement.

Objective To explore the effects of Chinese herbal compound Tangbikang on the expressions of p38 MAPK of sciatic nerve and plasma TNF-α in diabetic rats. Methods Ten of the sixty male SD rats were selected randomly as normal group, and the rest were fed with high-fat diet and low-dosage STZ was used to induce type Ⅱdiabetic rat models. Model rats were randomly divided into model group, mecobalamine group and Tangbikang low-, medium-, and high-dosage groups, 10 rats in each group. Each medication group was intervened with relevant medicine. Rat unilaterals sciatic nerves were taken after 16 weeks. The content of TNF-α in plasma was determined by radioimmunoassay. Western blot method was used to detect the expressions of p38 and p-p38 MAPK protein of sciatic nerve. Results Compared with normal group, the expressions of p38 and p-p38 protein and content of TNF-αin model group significantly increase (P<0.05, P<0.01). Compared with the model group, the expressions of p-p38 protein and the content of TNF-α significantly decreased after medicine intervention in different doses Tangbikang groups and mecobalamin group (P<0.05, P<0.01). The expression of p38 protein in Tangbikang high-dose group significantly decreased (P<0.05), with statistical significance (P<0.05). Conclusion Tangbikang can reduce the expression of p38 and p-p38 MAPK protein of the rat sciatic nerve, and reduce the content of TNF-α protein in rat plasma, which may be one of the effective targets of neuroprotection and abirritation of diabetic peripheral neuropathy.

Through the ethoxylation structure modification of three isoflavones which were genistein, biochanin A and formononetin in chickpea, the hypoglycemic activity and the synergistic hypoglycemic activity were studied. Ethoxylation structure modification was given on three kinds of isoflavones. The hypoglycemic activity of three kinds of isoflavones and their derivatives were studied. The synergistic hypoglycemic activity of the compounds was also studied. The insulin-resistance HepG2 cell was selected as the model of hypoglycemic activity screening. The results showed that four ethoxylation products were synthesized. And the hypoglycemic activity of genistein was better than biochanin A and formononetin with significant difference (P < 0.05). The maximum dosage of three isoflavones was compared with the positive drug metformin hydrochloride. And the effect was not as good as metformin hydrochloride. There was significant difference (P < 0.05, orP < 0.01). There was no significant difference of effect between the compound a and the compound d (P > 0.05). Effects of the compound b and compound c were not as good as the compound a and compound d. There was statistical difference (P < 0.05). The activity of compound d was significantly improved than the parent compound (P < 0.05). There was no statistical difference on the hypoglycemic activity between combination 5 and metformin hydrochloride (P > 0.05). It was concluded that the combinations of compounds played synergistic effects with better hypoglycemic effect. It provided a basis for developing hypoglycemic agents with the intellectual property rights.

This article was aimed to study the content determination methods ofTang-Nai-Kang (TNK) granules. The content of rosemary acid in TNK granules was determined by HPLC method, using C18 chromatographic column (150 mm×4.6 mm, 5μm), the mobile phase of 0.1% trifluoroacetic acid - methanol (60:40),λ of 330 nm. The theoretical plate number was more than 2 000. The standard curve method was used in the determination. Contents of ginsenosides Rg1, Re and Rb1 were determined by HPLC method, using C18 chromatographic column (150 mm× 4.6 mm, 5μm), the mobile phase of water-acetonitrile, gradient elution method,λ of 203 nm. The theoretical plate number was more than 2 000. The standard curve method was used in the determination. The results showed that rosmarinic acid was linear in the range of 0.300-0.500 mg·mL-1 with good linearity. The regression equation wasY = 1E + 07X -7 334,R2 = 0.999. Its sample recovery was 97.32% (RSD 1.25%). Ginsenoside Rg1 was linear in the range of 0.138-0.220 mg·mL-1 with good linearity. The regression equation wasY = 2E + 06X +34 864,R2 = 0. 999. Ginsenoside Re was linear in the range of 0.126-0.250 mg·mL-1 with good linearity. The regression equation wasY = 2E + 06X + 39 879,R2 = 0. 999. Ginsenoside Rb1 was linear in the range of 0.132-0.220 mg·mL-1 with good linearity. The regression equation wasY = 2E + 06X + 39 070,R2 =0. 999. Their sample recoveries were Rg1 97.97% (RSD 1.83%), Re 101.80% (RSD 1.83%), Rb1 98.35% (RSD 1.82%). It was concluded that the content determination method established for the quality control of TNK granules was simple, fast and reliable.

Spectrophotometry and single factor test were used in the investigation of effects on extraction results of total flavonoids from the fruit ofVernonia esculenta Hemsl. from aspects of ethanol concentration, frequency of extraction, length of extraction time, and the solid-liquid ratio. The extraction conditions were optimized by orthogonal test. The results showed that the optimum extraction conditions were ethanol concentration of 70%, extracted for 4 times, 1 h for each time, and the solid-liquid ratio of 1:20. Under these extraction conditions, the content of total flavonoids in the fruit ofVernonia esculenta Hemsl. was 103.55 mg·g-1.