OBJECTIVE： To develop a method of thin－ layer chromatographic scanning to determine the content of puerarin in Jiangtang pill METHODS： Silica GF254 thin－ layer was used， and the mobile phase consisted of chloroform－ methanol－ water(7∶ 2 5∶ 0 25) with the detection wave－ length of 254nm and the control wave－ length of 370nm RESULTS： The linear range of puerarin was 0 4～ 2 0μ g The mean recovery was 99 40％ (n=5) with RSD of 1 37％  CONCLUSION： This method is accurate， reliable and can be used for the determination of puerarin in Jiangtang pill

Objective To identify a high affinity IRF-1 binding element in the translational start site of XAF1 promoter. Methods By the bioinformatics analysis, a putative IFN regulatory factor 1 binding element (IRF-E), named as IRFE-XAF1, was identified from -30nt to -38nt of the XAF1 gene, with 76.2% homogeneity with the synonymous IRF-E sequence. Electrophoresis mobility shift assay (EMSA) was performed to confirm the binding capacity of IRFE-XAF1. Two site-directed mutations were made, one mutation site was outside of the IRF-E region (-28nt) and another located at the center of IRF-E (-34nt). The promoter of XAF1 after mutation was examined, including its binding activity and response to IFN-?. Results It was found by EMSA assay that the doublestranded oligonucleotide DNA probe, containing IRFE-XAF1 and labeled with 32P, may be connected to the nuclear protein, and blocked by the unlabeled synonymous IRF-1 probe (cold probe). The binding capacity of IRF-E was lost after site-directed mutation. The XAF1 promoter containing IRFE-XAF1 site had the priming activity, and could be induced by IFN-?. The priming activity declined markedly after site-directed mutation of IRFE-XAF1, and -34 site mutation completely eliminated the effect of IFN-?. Conclusion A high affinity of IRF-E is found in -30nt to -38nt region upstream of ATG initiator codon of XAF1 gene. The code sequence is -38nt-GAAACGAAA--30nt. The present study suggests that XAF1 is one of the genes with which IFN-? may induce the differentiation of cancer cells.

Objective:To investigate the possible role of long non-coding RNA (LncRNA) 00602 in promoting browning in adipocytes induced by adenovirus type 36 (Ad36).Methods:According to Ad36 infection, adipose tissue samples of obese patients were divided into Ad36-negative group and Ad36-infected group. Realtime fluorescent quantitative PCR (qRT-PCR) was used to detect the changes in the expression of LncRNA00602 mRNA in omental adipose tissue of the two groups, and analyze the differences between the two groups. The correlation between waist-to-hip ratio, systolic blood pressure, diastolic blood pressure, fasting blood glucose, triacylglyceride and other indicators of the patients in the group with LncRNA00602 mRNA expression were analyzed. HE staining was used to detect the size of adipocytes in the omental adipose tissue of the Ad36 negative group and the Ad36 infection group. qRT-PCR and Western blotting were used to detect the mRNA and protein expression levels of uncoupling protein 1 (UCP1) and PR domain containing 16 (PRDM16) in omental adipose tissue of two groups of patients. Human adipose-derived stem cells (hADSC) were isolated and cultured, using Ad36 to induce differentiation, and divided into control group and LncRNA00602 knockdown group. On 0, 2, and 4 days after LncRNA00602 knockdown, fluoroboron dipyrrole (BODIPY) and mitochondrial red fluorescence (Mito-Tracker Red) were used to stain intracellular lipid droplets and mitochondria. At the same time, qRT-PCR and Western blotting were used to detect changes in the expression of UCP1 and PRDM16.Results:The expression of LncRNA00602 gene in the Ad36 infection group was higher than that in the Ad36 negative group (all P<0.05). The expression of LncRNA00602 in the Ad36 negative group was not significantly different from the above clinical indicators, while the expression of LncRNA00602 was negatively correlated with serum fasting blood glucose and triacylglyceride ( r=-0.522, -0.486, P<0.05) in the Ad36 infection group; HE staining showed that the average adipocyte area of the Ad36 infection group was smaller than that of the Ad36 negative group. At the same time, UCP1 and PRDM16 gene expression were higher than the negative group (all P<0.05). At the cellular level, on the 2nd and 4th days after knockdown of LncRNA00602, the lipid droplet area of adipocytes in the LncRNA00602 knockdown group was larger than that of the control group, the number of mitochondria decreased compared with the control group, and difference was statistically significant ( P<0.05 or P<0.01); Compared with the control group, there was significantly lower expression of the browning marker genes UCP1, PRDM16, and protein in the adipocytes in the LncRNA00602 knockdown group (all P<0.05). Conclusion:In Ad36-induced adipocyte differentiation, LncRNA00602 may positively regulate the expression of UCP1, PRDM16 and lipid droplet metabolism, and promote the browning of adipocytes.