OBJECTIVE:To observe the correlation of concentration-time curves of riboflavin in salive and plasma.METHOD:After peple orally taking riboflavin tablets,the riboflavin in salive was determinated by fluorescence spectrophotometry with excitation wavelength 475.4nm and emission wavelength 524.7nm.RESULTS:The recovery of riboflavin was over 96%.The detection limits of riboflavin was 5ng.ml-1,and the calibration curve was linear in the range of 5ng.ml-1～3μg.ml-1.The regression equation was △F′=0.116c+0.232(r=0.9999).CONCLUSION:The concentration-time curves of riboflavin in salive was similar to these in plasma.

Objective:To investigate the expression of TAZ and KLF5 and their clinical significance in hepatocellular carcinoma ( HCC).Methods:We freshly collected 76 samples of surgically resected HCC and matched normal tumor-adjacent tissues and detected TAZ and KLF5 expression in these samples using immunohistochemical staining.The clinical significance of TAZ and KLF5 protein expression were analysed.Results:The protein expression of TAZ and KLF5 in HCC tissues was significantly higher than those in matched normal tumor-adjacent tissues ( P=0.001;P=0.035 ).Clinicopathological analysis suggested that TAZ and KLF5 protein expression were associated with histopathological differentiation ( P=0.007;P=0.047 ) and TNM stage ( P=0.009;P=0.040).TAZ was positively correlated with KLF5 protein in HCC tissues (r=0.651,P=0.003).Conclusion:The high-expression of TAZ and KLF5 are correlated with poor clinicopathological characteristics,and TAZ is positively associated with KLF5 in HCC tissues, suggesting that TAZ may promote tumor progression through inhibition of KLF5 protein degradation in HCC.

Objective To construct a cDNA expression library from unfed female tick Haemaphysalis longicornis for screening and cloning potential antigenic genes.Methods Total RNA was isolated from unfed female ticks,mRNA was purified and a library of oligo(dT)-primed cDNA with added directional EcoR Ⅰ/Hind Ⅲ linkers was constructed from the purified mRNA.The constructed cDNA was ligated to the EcoRⅠ/HindⅢ arms of the ?SCREEN vector.Pure phage stocks were harvested by plaque purification and converted to plasmid subclones by plating phage on host strain BM25.8.Recombinant plasmids that were subcloned to E.coli BM25.8 were isolated and transformed into E.coli JM109.Recombinant plasmids abstracted from JM109 were identified by PCR and sequencing.Rusults The recombinant phage DNA was packaged by using phage-marker packaging extracts,resulting in a primary cDNA library with a size of 1.8?106 pfu.Data showed 100% of the library were recombinant and the titer of the amplified library was 2.4?109 pfu/ml.Forty-two clones of encoding immunodominant antigens were obtained from the cDNA library.Sequence analysis revealed 12 unique cDNA sequences and the encoded putative proteins showed similarities to H.longicornis tropomyosin mRNA,Rhipicephalus annulatus unknown larval protein mRNA,chromosome 2R of Drosophila melanogaster,mitochondrial DNA of H.flava,clones HqL09 unkown mRNA and Hq05 mRNA of H.qinghaiensis,and myosin alkali light chain protein mRNA.Conclusion The cDNA expression library from unfed female H.longicornis was successfully constructed and screening of protective genes may provide candidate antigens of the tick.

AIM:To investigate the effects of Prunus cerasifera Ehrh.polyphenol extract(PCE)on glucose metabolism in insulin-resistant HepG2(IR-HepG2)cells.METHODS:An IR-HepG2 cells model was established using high sugar and high insulin induction;a control group,an insulin resistance(IR)model group,a PCE treatment group with different concentrations,and a metformin treatment group were set up.Cells were treated with 50,100,200,and 400 mg/L of polyphenol extract for 24 hours,respectively,to detect their effects on cellular glucose consumption.At the same time,use thiazole blue(MTT)colorimetry to evaluate cell viability.Real-time PCR and Western blot were used to detect expression of glucose metabolism-related mRNA and proteins.RESULTS:Prunus cerasifera Ehrh.polyphenol ex-tract can significantly increase glucose consumption in IR-HepG2 cells,and cell activity tends to increase and then decline as its concentration increases;At mRNA and protein levels,in addition to significantly increasing the expression of phos-phorylated adenosine monophosphate activated protein kinase(p-AMPK)protein in IR-HepG2 cells,it also decrease gly-cogen synthase kinase 3β(GSK3β)in glycogen synthesis and gluconeogenesis pathways,forked protein O1(FoxO1),peroxisome proliferator activates gamma co-receptor activation 1-alpha(PGC-1α),glucose-6-phosphatase(G6Pase)and phosphoenol pyruvate carboxykinase(PEPCK)expression.CONCLUSION:Prunus cerasifera Ehrh.polyphenol extract can activate the AMPK signaling pathway in IR-HepG2 cells,to further inhibit gluconeogenesis and glycogen synthesis pathways,thereby improving the insulin resistance state of HepG2 cells.