Objective:To establish the quality standard for Qiyeling granules. Methods:Astragali Radix and naringin were identi-fied by TLC. The content of astragaloside A was determined by HPLC. Results:The qualitative identification was with high resolution and without interference from the negative substances. The linear range of astragaloside A was 0. 166-1. 104 μg(r=0. 999 1), and the average recovery was 97. 27%(RSD=1. 3%,n=6). Conclusion:The method can be used in the quality control of Qiyeling granules.

Objective: To observe the effect of a novel targeted agent enzastaurin alone or in combination with gefitinib on ge-fitinib-resistant human non-small cell lung cancer cells to explore the rational drug combination. Methods: CCK-8 assay was used to measure the cell proliferation. Combination index ( CI) was calculated by Chou-Talalay method to assess the efficacy of the combination therapy. The flow cytometry (FCM) was used to analyze the change in the cell cycle. Results:In 72h, the IC50 value of gefitinib and enzastaurin for the lung cancer NCI-H460 cells was 6. 99μmol·L-1 (95%CI:3. 55-13. 79μmol·L-1 ) and 7. 25μmol·L-1 (95%CI:4. 77-1. 02 μmol·L-1), respectively. The inhibition effect on the cell proliferation was stronger in the combination treatment than that in the monotherapy (P<0. 05), and the simultaneous treatment showed the most significant inhibition effect (P<0. 01). The IC50 value of gefitinib for H460 cells in the simultaneous administration group, the sequential administration group with gefitinib used first and the sequential administration group with enzastaurin used first was 0.006 μmol·L-1(95%CI:0.002-0.020μmol·L-1), 0.02μmol·L-1(95%CI:0.011-0.037 μmol·L-1) and 0.085 μmol·L-1(95% CI:0.042-0.170μmol·L-1, respectively. The CI of the simultaneous administration group was lower than one when the gefitinib concentration was above 0. 05μmol·L-1 . The cell cycle distribution result indicated that the simultaneous administration group had significantly increased G0/G1 proportion (P<0. 05) and induced cell cycle arrest at G1 phase. Conclusion:Protein kinase C inhibitor enzastaurin combined with EGFR inhibitor gefitinib shows a synergistic effect, suggesting that the combination treatment of the two drugs might be a new strategy for the follow-up therapy of gefitinib-resistant non-small cell lung cancer.

BACKGROUNDTo observe the suppressive effects of exogenous p27 gene on human lung cancer cell line A549.

METHODSAn adenovirus expression vector (pAd CMV-p27) containing 570 bp human full-length p27 cDNA was transfected into human lung cancer cell line A549. Expression of exogenous p27 gene was detected by dot-blot hybridization and laser co-focal system. MTT was adopted to measure the effects of exogenous p27 gene on cell cycle progression and cell features of the infected A549.

RESULTSThe mRNA and protein expression level of p27 was remarkably increased after transfecting with exogenous p27 gene. The apoptosis of infected A549 occurred and the progression of cell cycle was arrested in G1 phase.

CONCLUSIONSp27(kipl) gene transfer may play a therapeutic role in the treatment of lung cancer.

BACKGROUNDTo investigate the biological effects of anti VEGF₁₆₅ ribozyme on human lung adenocarcinoma cell.

METHODSHammerhead ribozyme (VRz) against VEGF₁₆₅ gene transcripts (site 212) and its paired mutant ribozyme (mVRz) were designed and synthesized, and the cleavage activity of the ribozymes on target RNA in a cell-free system was observed. The replication-incompetent adenovirus-mediated eukaryotic expression vectors (rpAdVRz) containing VRz and mVRz gene were constructed and identified. Then the human lung adenocarcinoma cells (A549) were infected with recombinant adenovirus. The biological characteristics of A549 cell before and after infection in vitro were inspected by Northern blot, laser confocal imaging system analysis, flow cytometry and transmission electron microscopy.

RESULTSVRz specifically and efficiently cleaved the VEGF₁₆₅ mRNA. The rpAdVRz was successfully constructed and infected A549 cell. The level of VEGF₁₆₅ expression decreased 87% in rpAdVRz infected cells compared with the other groups, but their biological characteristics were not influenced by the expression of the exogenous gene.

CONCLUSIONSThe adenovirus mediated hammerhead ribozyme against VEGF₁₆₅ can significantly decrease the expression of VEGF₁₆₅. This provides an experimental basis for human lung cancer gene therapy with antiangiogenesis method.

Objective:To explore the protective effects and related mechanism of tea saponin (TS) on intestinal inflammation due to Shigella infection. Methods:In vitro, the antibacterial activity of TS was detected by standard broth microdilution method. The absorbance at 600 nm of the bacterial liquid was detected by Microplate Reader under different concentrations of TS, and the growth curve was drawn. Bacterial count was obtained by plate colony counting. In vivo, 15 mice were divided into 3 groups ( n=5 each): control group, Sf301 group and TS group. Sterile water or TS was applied to mice in the Sf301 group and the TS group per gavage once a day for 8 days, and the mouse model of Shigella infection was established on day 3. The disease activity index (DAI) was used to evaluate the general condition of mice. The mice were sacrificed on day 8. Colon length was measured and colon tissues were stained with HE to analyze the pathological changes. The cecal contents and feces of mice were taken for plate counting and Shigella load was obtained. Real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the mRNA expression levels of inflammatory factors. Results:(1) In vitro, the MIC of TS was 1 024 μg/ml. The plate counting and A600 of TS decreased in proportion to increasing concentrations (256, 512, 1 024 μg/ml) in comparison with the control group (all P<0.05). (2) In vivo, colon length was (7.70±0.24) cm in the control group and (7.35±0.41) cm in the TS group, which was significantly longer than that of the Sf301 group ([6.13±0.05] cm, P<0.05). Histopathological examination evidenced colonic epithelial cells shedding, decreased goblet cells, and inflammatory cell infiltration in the colon tissue of the Sf301 group. In the TS group, the colonic mucosa was intact without significant inflammatory cell infiltration. Bacterial load in cecal contents was significantly lower in the TS group than in the Sf301 group ( P<0.05). The level of tumor necrosis factor-α was significantly lower, and the level of interleukin-10 was significantly higher in the TS group than in the Sf301 group (all P<0.05). Conclusion:TS can effectively inhibit Shigella and alleviate Shigella infective enteritis by reducing Shigella load and inhibiting inflammation.

Low-carbohydrate diet is becoming popular recently. It requires carbohydrate intake lower than 130 g/d(or <26% of total energy intake) while fat and protein intake are not defined. Low-carbohydrate diet contributes to better glycemic control in patients with type 1 diabetes: lowering HbA 1C and glycemic variability, and improving time in range(TIR). Low-carbohydrate diet reduces insulin dosage in type 1 diabetic patients and does not increase the risks of hypoglycemia and diabetic ketoacidosis. This review summarized the research evidence of low-carbohydrate diet to explore the role and safety in disease management.