Objective:To study the effect of the cytokines, tumor necrosis factor-?(TNF-?), interferon-?(IFN-?), interferon-?(IFN-?), platelet-derived growth factor BB(PDGF-BB), basic fibroblast growth factor(bFGF),on promoter activity of human transforming growth factor-?1(TGF-?1) gene.Methods:A construct of phTGF2.14 containing sequence from -1 328 bp to +812 bp of human TGF-?1 gene,linking with chloramphenicol acetyltransferase(CAT) as reporter gene,was transiently transfected into rat hepatic stellate cell line nFSC. The cells were subsequently treated with TNF-?,IFN-?,IFN-?,PDGF-BB,bFGF, and the CAT activity was assessed 48 hours after stimulation with each cytokines.Results:TNF-? of 10 ng/ml can increased the CAT activity of phTGF2.14 to 3.24 fold compared to control. IFN-? and IFN-? at 1 000 U/ml decreased CAT activity to (42?12)% and (58?6)% of control respectively.PDGF-BB,bFGF of 10 ng/ml had no effect on promoter activity of human TGF-?1 gene;Combined application of IFN-?,IFN-? and TNF-?,the promoter TGF-?1 gene were 1.32 and 1.46 fold compared with control,respectively.Conclusion:These data indicate that TNF-? can increase the promoter activity of human TGF-?1 gene, but IFN-?,IFN? can downregulate the CAT activites of phTGF2.14, and IFN-?,IFN-? can interdict the upregulate effect of TNF-? on phTGF2.14. We did not find PDGF-BB,bFGF have any effect on TGF-?1 promoter. These provided an essential evidence for study the interaction mechanism of cytokines in fibrogenisis diseases.

Objective:To investigate the effects of TGF-?1 on promoter activity of human transforming growth factor-beta1(TGF-?1) gene.Methods:The fragments with different length of the 5′-end sequence of the human TGF-?1 gene between -1 328 bp to +812 bp were obtained from a healthy male person genomic DNA, and the sequences were fused to chloramphenicol acetyltransferase reporter gene in the pCAT-enhancer plasmid to construct five chimeric recombinant plasmids. These recombinant plasmids were transiently transfected into rat hepatic setellate cell line(nFSC) by FuGENE6 transfection methods. The cells transfected with chimeric recombinant plasmids were cultured on media in the absence or presence of TGF-?1(2 5 ng/ml),CAT activities in transfected cells were tested and compared.Results:The different concentration of TGF-?1 can prevent the proliferation of nFSC, and these effects are dose-dependent; The presence TGF-?1 can stimulate the CAT activity of cells transfected with phTGF0 585?phTGF1 120?phTGF1 423?phTGF1 680?phTGF2 140 up to 2～5 times higher then cells cultured with absence of TGF-?1.Conclusion:The TGF-?1 can stimulate CAT activities in nFSC transfected with phTGF0 585?phTGF1 120?phTGF1 423?phTGF1 680?phTGF2 140. These results suggest that TGF-?1 have an autoregulation effect on TGF-?1 gene.

Objective To evaluate the image quality,diagnosis accuracy and dose reduction of split-bolus CT urography (CTU) with low voltage scan and sinogram affirmed iterative reconstruction (SAFIRE).Methods A total of 80 cases of consecutive patients with confirmed or suspected urinary system disease needed CTU examination were divided into two groups (control group and test group) by using random number table.In control group,convention scan (120 kV) with one time injection was used.But low voltage scan (80 kV) with SAFIRE algorithm and split-bolus injection (SBI) was used in experiment group.The radiation dose,image quality and diagnosis accuracy were compared.Results A total of 77 cases completed CTU examination successfully in the two groups,including 39 cases in control group and 38 cases in test group.The effective dose reduced from (26.68 ± 4.07) in control group to (3.93 ± 0.85) mSv in test group (t =-33.78,P ＜ 0.05).Subjective image quality score was (4.49 ± 0.79) in control group and (4.39 ± 1.53) in test group,with no significantly statistical difference (Z =2.71,P ＞ 0.05).Signal-to-noise ratio (SNR) of objective image quality in test group was higher than that in control group (127.3±15.9 vs.109.6 ± 13.2,t =4.49,P＜0.05).But there was no significantly statistical difference in contrast-to-noise ratio (CNR) between control group(100.8 ± 12.9)and test group (109.0 ± 14.4,P ＞ 0.05).For diagnosis accuracy,no statistical difference were found between two groups(84.62％ and 81.58％,P ＞ 0.05).Conclusions The combination of low voltage scan with SAFIRE algorithm and split-bolus injection CTU could reduce the radiation dose significantly,but the objective image quality,CNR (except SNR) of subjective image quality and diagnosis accuracy were all unaffected obviously.

Objective To explore the effects and mechanisms of hepatitis B virus-X protein (HBx) on invasion and migration of hepatocellular carcinoma (HCC) cells.Methods The retrospective cohort study was conducted.The clinicopathological data of 30 patients with liver tumor (20 with HCC and 10 with benign tumor of liver) who were admitted to the Affiliated Hospital of Xuzhou Medical College between July 2014 and July 2015 were collected.HCC tissues of 20 patients with HCC (with history of HBV infection) were collected by surgical resection and peritumoral normal tissues (outside of tumor capsule) of 10 patients with benign tumor of liver (without history of HBV infection) were collected.The expressions of epidermal growth factor receptor 3 (ErbB3)in HCC tissues and peritumoral normal tissues were detected by immunohistochemistry (IHC).The relative expressions of ErbB3 and HBx in HCC tissues and peritumoral normal tissues were detected by Western blot,and relative expressions of ErbB3 in HepG2 of which green fluorescent protein (GFP) and GFP-HBx were respectively transfected were detected.The relative expressions of ErbB3 mRNA in HepG2 transfected by GFP and GFP-HBx were detected by real-time polymerase chain reaction (RT-PCR).The migration and invasion of HepG2 were respectively detected by Transwell assay with and without matrix.The measurement data with normal distribution were represented as $± s.The comparisons between groups were evaluated with the independent-sample t test.Correlation analysis was done by the Pearson test.Results (1) The expressions of ErbB3 were detected by IHC:relative value of mean optical density (MOD) of ErbB3 in HCC tissues of 20 patients with HCC and peritumoral normal tissues of 10 patients with benign tumor of liver were 2.54± 1.33 and O.99±0.29,respectively,with a statistically significant difference (t =6.542,P ＜ 0.05).(2) The relative expressions of ErbB3 and HBx were detected by Western blot:relative expressions of ErbB3 and HBx were respectively 0.79±0.13,1.10±0.28 in HCC tissues of 10 patients with HCC and 1.07±0.17,0 in peritumoral normal tissues of 10 patients with benign tumor of liver,with statistically significant differences (t =3.229,19.486,P＜0.05).The results of Pearson test showed that there was a positive correlation of expression between ErbB3 and HBx in HCC tissues (r=O.637,P＜ 0.05).(3) The relative expressions and transcriptional levels of ErbB3 were detected by Western blot and RT-PCR:relative expressions of ErbB3 in HepG2 of which GFP and GFP-HBx were respectively transfected were O.75±0.11 and 1.10±0.10,respectively,with a statistically significant difference (t=4.291,P＜0.05).The relative expressions of ErbB3 mRNA in HepG2 of which GFP and GFP-HBx were respectively transfected were O.38±0.03 and O.94±0.07,respectively,with a statistically significant difference (t=11.703,P＜O.05).(4) The effects of ErbB3 on migration and invasion of HepG2:numbers of transmenbrane cell in HepG2 of which His and His-ErbB3 were respectively transfected by Transwell assay with matrix were respectively 271± 18 and 463± 31,respectively,with a statistically significant difference (t =8.202,P＜0.05).Numbers of transmenbrane cell in HepG2 of which His and His-ErbB3 were respectively transfected by Transwell assay without matrix were respectively 315±38 and 549±34,respectively,with a statistically significant difference (t =8.310,P＜0.05).Conclusion HBx protein can promote the invasion and migration of hepatocellular carcinoma cells through up-regulating expressions of ErbB3 protein.

This study was intended to establish a method of purification of HPV16 L1 protein expressed in a prokaryotic system and to obtain the purified protein. The prokaryotic expression vector pGEX-4T-1-HPV16 L1 was constructed and transformed into E. coli BL21 cell, and induced by 1 mM IPTG to express HPV16L1 protein. The inclusion bodies were isolated and solubilized with 8 M urea. After the urea was removed by gradual dialysis, the denatured L1 protein were renatured and then were purified by affinity chromatography. The results showed that HPV16L1 protein formed inclusion bodies in bacterial expression system, suggesting that this assay can be used to purify HPV16L1 protein and hence provide a basis for studying the applications of HPV16 L1 protein.
Capsid Proteins ; biosynthesis ; Escherichia coli ; metabolism ; Genetic Vectors ; Human papillomavirus 16 ; Oncogene Proteins, Viral ; biosynthesis