Objective To establish an HPLC method for determination of pterodontic acid in Lingdancao Soft Capsula. Methods Agilent Zorbax XDB-C18 column (250 mm?4.6 mm, 5 ?m) was used. Acetonitrile-water (60∶40) was used as mobile phase, the detection wavelength was 210 nm, flow rate was 1.0 mL/min, and column temperature was at 25 ℃. Results The linear range of pterodontic acid was 12.91—129.1 ?g/mL (r=0.999 9), the average recovery rate of pterodontic acid was 99.7 %, RSD was 2.0%. Conclusion The method is simple, accurate, and reliable, and it can be used to the quality control of this preparation effectively as a quantitative method.

A novel method for quantification of astragaloside IV in Radix Astragli was developed by using online SPE liquid chromatography coupled with corona charged aerosol detection( CAD) . The sample solution
was loaded into Acclaim Polar AdvantageⅡC18(50 mm×4. 6 mm, 3 μm) which was selected as online SPE column. Then the cleaning process was done by using the Right one of dual gradient pumps with Methanol-water as mobile phase. Acclaim C18(150 mm×4. 6 mm, 5 μm) was selected as analytical column with acetonitrile-water as mobile phase at a flow rate of 1. 0 mL/min. The eluate containing the target from SPE column was transferred into the analytical column by heart-cutting mode. The temperature of nebulizer of corona CAD was set at 30 ℃ and the nitrogen pressure was 241. 3 kPa. The baseline separation of astragaloside IV from matrix components has been achieved. There was a good linear correlation in the range of 4 . 0-80 mg/L for astragaloside IV and the correlation coefficient was 0 . 9998 . The standard addition average recovery of astragaloside IV in Radix Astragli was 97 . 6%. It has been validated that astragaloside IV in Radix Astragli and its preparations can be quantified rapidly and accurately with this method.

Objective:The purpose of this study is to use the next-generation sequencing (NGS) technology platform to detect the methylation rate of phosphatase and tensin homolog deleted on chromosome ten ( PTEN) promoter region in hepatocellular carcinoma (HCC) tissue samples, and to analyze the clinical significance of its correlation with the prognosis of patients receiving sorafenib treatment. Methods:The 52 pairs of tumor tissue and para-cancerous tissue samples from HCC patients treated with sorafenib alone, which were collected and preserved in the Liver Tumor Diagnosis and Research Center of the former 302 Hospital of the People′s Liberation Army by the National Natural Science Foundation of China Youth Project with the project batch number 81702986 in 2018, were extracted total DNA from the samples. Then the DNA samples were treated with bisulfite and specific primers were designed to amplify the PTEN promoter region. Finally, the amplified products were analyzed by second-generation sequencing. In the analysis of clinical significance of PTEN methylation, log-rank statistical analysis was used to calculate whether there was a statistical difference in survival between the patient groups. Results:The methylation rate of PTEN promoter region in tumor tissues (29.17%±9.58%) was significantly higher than that in paracancer tissues (4.17%±2.86%)( t=19.970, P<0.05). At the same time, in HCC tissues, the methylation rate of the PTEN promoter region is negatively correlated with its expression ( F=47.270, P<0.000 1; Y=-1 800× X+38.03), and the PTEN methylation rate is negatively correlated with the prognosis of patients receiving the molecularly targeted drug Sorafenib (χ2=4.313, P<0.05). Conclusion:This study successfully established a new method for detecting methylation in the promoter region of PTEN, and the methylation rate of PTEN can be used as one of the targets of HCC diagnosis and targeted therapy.

Objective:The purpose of this study is to use the next-generation sequencing (NGS) technology platform to detect the methylation rate of phosphatase and tensin homolog deleted on chromosome ten ( PTEN) promoter region in hepatocellular carcinoma (HCC) tissue samples, and to analyze the clinical significance of its correlation with the prognosis of patients receiving sorafenib treatment. Methods:The 52 pairs of tumor tissue and para-cancerous tissue samples from HCC patients treated with sorafenib alone, which were collected and preserved in the Liver Tumor Diagnosis and Research Center of the former 302 Hospital of the People′s Liberation Army by the National Natural Science Foundation of China Youth Project with the project batch number 81702986 in 2018, were extracted total DNA from the samples. Then the DNA samples were treated with bisulfite and specific primers were designed to amplify the PTEN promoter region. Finally, the amplified products were analyzed by second-generation sequencing. In the analysis of clinical significance of PTEN methylation, log-rank statistical analysis was used to calculate whether there was a statistical difference in survival between the patient groups. Results:The methylation rate of PTEN promoter region in tumor tissues (29.17%±9.58%) was significantly higher than that in paracancer tissues (4.17%±2.86%)( t=19.970, P<0.05). At the same time, in HCC tissues, the methylation rate of the PTEN promoter region is negatively correlated with its expression ( F=47.270, P<0.000 1; Y=-1 800× X+38.03), and the PTEN methylation rate is negatively correlated with the prognosis of patients receiving the molecularly targeted drug Sorafenib (χ2=4.313, P<0.05). Conclusion:This study successfully established a new method for detecting methylation in the promoter region of PTEN, and the methylation rate of PTEN can be used as one of the targets of HCC diagnosis and targeted therapy.