PCR-RFLP for Distinguishing Periplocae Cortex from Acanthopanacis Cortex and Lycii Cortex / 中国实验方剂学杂志

ObjectiveTo establish a polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） method for rapid distinguishing Periplocae Cortex from Acanthopanacis Cortex and Lycii Cortex， so as to avoid the influence of genetic confusion on drug safety. MethodThe DSS-tagged sequences of Periplocae Cortex were obtained from the Chloroplast Genome Information Resource （CGIR） and analyzed to find the enzymatic cleavage sites that were different from those of Acanthopanacis Cortex and Lycii Cortex. The specific enzymatic cleavage site， Cla I， of Periplocae Cortex was selected， on the basis of which the primers for PCR-RFLP were designed. Furthermore， the factors such as annealing temperature， number of cycles， Taq enzyme， PCR instruments， and enzymatic treatment time that may influence PCR-RFLP were studied. The established PCR-RFLP method was applied to the identification of Periplocae Cortex， Acanthopanacis Cortex， and Lycii Cortex samples produced in different regions. ResultThe PCR-RFLP at the annealing temperature of 59 ℃ and with 40 cycles showed clear bands of the samples. When the enzyme digestion time was 30 min. The reaction produced the target bands at about 140 bp and 290 bp for both Periplocae Cortex and its original plant and only a band at about 430 bp for Acanthopanacis Cortex， Lycii Cortex， and their original plants. The method can accurately distinguish Periplocae Cortex from its confounders Acanthopanacis Cortex and Lycii Cortex. ConclusionThe PCR-RFLP method for distinguishing Periplocae Cortex from Acanthopanacis Cortex and Lycii Cortex was established. It has high stability， sensitivity， and applicability， providing a reference for the quality control of Periplocae Cortex， Acanthopanacis Cortex， and Lycii Cortex.

Androgen receptor up-regulates EphA3 expression in androgen-dependent prostate cancer cell lines / 天津医药

Objective To evaluate the relationship between liver cell type A receptor (EphA) expression and androgen receptor (AR) signaling in androgen-dependent prostate cancer cells. Methods RT-PCR and Western blot assay were used to determine mRNA and protein levels of EphA3 and AR in prostate cancer LNCaP and 22Rv1 cells, respectively. The variations of EphA3, AR and prostate specific antigen (PSA) expressions were also measured in these cells after dihydrotes?tosterone (DHT) treatment for 48 h. The constructed EphA3-Luc (-789-+146) luciferase reporter plasmid was co-transfect?ed with pcDNA3.1(+)-AR or siAR in 22Rv1 cells to analyze the effects of different AR expression levels on EphA3 tran?scription activity. Results The expression pattern of EphA3 was similar to AR, showing a lower level in prostate stromal cell line WPMY-1 and a higher level in prostate cancer cell lines LNCaP and 22Rv1. When stimulated with 10 nmol/L DHT, the expression levels of AR, PSA and EphA3 were significantly increased in 22Rv1 cells, and the protein levels of these genes were also increased in LNCaP cells. Moreover, AR expression levels markedly influenced the activity of EphA 3 pro?moter. Conclusion AR up-regulates EphA3 expression by increasing the activity of EphA3 promoter.