Résumé
Objective To analyze the relationship among serum hepatitis B virus (HBV ) envelope large protein (HBV‐LP ) , HBV‐DNA and HBV marker(HBV‐M ) for investigating the clinical significance of HBV‐LP to reflect the HBV in vivo replication in the patients with HBV infection .Methods Total 540 cases of chronic HBV infection treated in the Longgang District Hospital of Traditional Chinese Medicine from April 2013 to September 2015 were selected .The real‐time fluorescence quantitative PCR meth‐od was used to detect serum HBV‐DNA ,HBV‐LP and HBV‐M were detected by the enzyme linked immunosorbent assay (ELISA) . The correlation among HBV‐LP ,HBV‐M and HBV‐DNA were analyzed .Results The positive rate of HBV‐LP in HBeAg‐positive patients was 96 .39% ,and which of HBV‐DNA was 93 .33% ,there was no statistically significant difference between them (P>0 .05);The serum HBV‐LP level was positively related with the logarithmic value of HBV‐DNA copies ;the positive rate of HBV‐LP in HBeAg‐negative patients was 63 .33% ,and which of HBV‐DNA was 51 .11% ,the difference between them was statistically significant(P<0 .05) .Conclusion HBV‐LP can effectively reflect the HBV in vivo replication in the patients with chronic hepatitis B and its sensitivity is higher than that of HBeAg ,HBV‐LP can even more reflect the HBV in vivo replication status in patients with HBeAg‐negative chronic hepatitis B .
Résumé
Objective: To construct a liver targeting gene transfer vector using hepatitis B virus envelope particles. Methods: Hepatitis B viruses were obtained from the supernatant of HepG 2.2.15 cells by a PEG8000 system and were inactivated by ?-propiolactone to prepare hepatitis B virus envelope. The hepatitis B virus envelope was used to pack 5.3 kb pIRES_2-EGFP to assess their packing ability. Subsequently, the products were studied with ELISA, PCR, SDS-PAGE, and electron microscopy. Finally, the product was used to transfect HepG2 cells and the green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results: The acquired hepatitis B virus envelope retained the surface protein HBsAg+pre S_1+pre S_2, but with no virus DNA. The prepared envelope had high packing ability for GFP and the packed GFP had a high transfection rate in HepG2 cell. Conclusion: Hepatitis B virus envelope has been successfully obtained from the supernatant of HepG 2.2.15 cells with a PEG8000 system and ?-propiolactone.
Résumé
Background and purpose:The reason for hepatitis B virus (HBV) with hepatocyte specificity is PreS1 enchased on the hepatitis B virus envelope (HBVE). So HBVE may have a potential application in liver targeting gene transfer. In this study, we investigated whether HBVE has the ability to target liver cancer cells. Methods:HBVE was obtained from the supernatant of Hep G 2.2.15 cells through PEG8000 system and ?-propiolactone method. The pIRS2-EGFP was packed with HBVE and resulted in the product HBVE-GFP. HBVE-GFP was transfected into HepG2, A549, HeLa and FB cells. The green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results:The GFP could be observed in the four groups, but the HepG2 group had a higher fluorescent intensity than the other 3 groups. The transfected rate of HepG2 group was (71.35?0.03)% , much higher than other groups(P
Résumé
Objective:To observe the transfection efficacy and targeting efficiency of hepatitis B virus envelope(HBVE) as a gene transfer vector for liver cancer cells.Methods:HBVE was obtained from the supernatant of HepG2.2.15 cells with a PEG8000 system and ?-propiolactone.The pIRS2-EGFP was packed with HBVE to obtain HBVE-GFP and was packed with liposome to obtain Liposome-GFP.HBVE-GFP and Liposome-GFP were used to transfect human hepatoblastoma cell line HepG 2 to study the transfection efficiency.HepG 2,A 549,HeLa and FB cells were transfected with HBVE-GFP to appraise the targeting ability of HBVE-GFP.GFP protein expression was observed under a fluorescent microscope and the ratio of GFP positive cells was determined by flow cytometry.Results:(1) Transfection efficiency:The GFP protein was observed in both the liposome group and the HBVE group under the fluorescent microscope;the fluorescent intensity in the HBVE group was 3-4 times that of liposome group as determined by flow cytometry(P