This study was aimed to screen candidate genes involved in the triterpenoid saponins biosynthetic pathway of the Ilex asprella root. The Illumina platform was applied to perform transcriptomic sequencing of I. asprella root, followed by a series of bioinformatics analysis. The results showed that a total of 272 candidate unigenes were anno-tated to be involved in the biosynthetic pathway of terpenoid in the transcriptome of I. asprella root, including 72 u-nigenes for the upstream pathway and 26 unigenes for cyclization, oxidation and glycosylation in the downstream pathway. Phylogenetic analysis was carried out to further analyze the evolution relationship of some candidate uni-genes and their homologous genes. Two genes IaA S1 and IaA S2 were proved to be mixed amyrin synthases in yeast expression system. Moreover, IaA S1 was identified to one of the rare ASs with α-amyrin as the major product. It was concluded that a series of candidate genes, which might be involved in the biosynthetic pathway of triterpenoid saponins, were screened out from the transcriptome of I. asprella root. Further investigation of these candidate genes will provide insight into their actual functions in the triterpenoid saponins biosynthetic pathway in I. asprella.

Objective To establish the quality standard of Radix Toddaliae Asiaticae. Methods Thin layer chromatography ( TLC) and high performance liquid chromatography ( HPLC) were used to identify and determine chloride nitidine and toddalolactone in Radix Toddaliae Asiaticae. The moisture and total ash contents were detected according to the methods recorded in appendix of Chinese Pharmacopeia (2010 edition) . Results Toddalolactone and chloride nitidine were detectable by TLC, the spots were clear and the dissociation was good. The established HPLC method was simple and accurate. The linear ranges of toddalolactone and chloride nitidine in Radix Toddaliae Asiaticae were 2.84~42.6 μg/mL and 25.6~385 μg/mL, and their recovery rates were 99.2 % ( RSD=1.12%) and 100 % ( RSD=0.71%) , respectively. The content of moisture was in the range of 75.8~98.9 mg/g and that of total ash was in the range of 12.4~33.6 mg/g. Conclusion The developed method is specific and accurate, and can provide useful reference for establishing quality standard of Radix Toddaliae Asiaticae.

Objective To optimize the high performance liquid chromatography (HPLC) method for determining the content of scopoletin from Caulis Erycibes. Methods Methanol-25% HCl ( v/v, 4 : 1) solvent was used to extract scopoletin. HPLC method was performed on Waters XBridge Shield RP18 column (4.6 mm × 250 mm, 5μm) with the mobile phase consisting of acetonitrile ( A) and 0.16% ( v/v) acetic acid ( B) solution by gradient elution. The detection wavelength was 298 nm and the flow rate was set at 1.0 mL/min. Results The linear range of scopoletin from Caulis Erycibes was 2.83-118 μg/mL, and the recovery rate was 99.47% ( sR=1.07%). Conclusion The optimized method is simple, specific and accurate, and can provide reference for content determination of scopoletin in Caulis Erycibes.

Objective The purpose of our study is to investigate the clinical value of virtual touch tissue quantification (VTQ)in the differential diagnosis of the benign and malignant liver tumor.Methods VTQ was used to inspect 53 patients with liver space-occupying lesions.Results After choosing the little lesions (the largest diameter less than 35 mm) from all the case groups,the VTQ value of the malignant group was higher than the hepatic hemangioma group (with significant differences among the different groups,P ＞0.05).Through the analysis of receiver operating characteristic curve (ROC),the optimal cutoff point as diagnosis reference value for differential diagnosis between malignant and benign groups was available.When the Area under the curve was 0.778,and with the VTQ value of 1.91 m/s,the sensitivity,specificity and Youden index was 78.6％,90.0％,0.686 respectively.The correlation analysis showed that the VTQ values of the lesion was positively correlated with the largest diameter of the lesion in the liver hemangioma group,with significant statistically differences (P ＜ 0.05).The correlation index was 0.812.Conclusions VTQ could provide a new effective way to benign and malignant liver lesions in differential diagnosis.