The Relationship Between The Quantities Of Hepatitis B Virus Covalently Closed Circular DNA And Lasting Time Of Serum Virology Response

[Objective]Covalently closed circular DNA (ccc DNA) is the original replication template of hepatitis B virus (HBV). The replication and existence of cccDNA maintains the source of infection. Suppression or removal of cccDNA is the key to the treatment of chronic hepatitis B. Only depend on a simple monitoring of liver function in peripheral blood and the level of HBV DNA is far from enough in the course of anti-hepatitis B virus treatment. The recurrence rate after stopping is still high even if one year continued treatment after the conversion of serum HBeAg happen in chronic hepatitis B patients. Anti-virus treatment, such as drug stopping target has become a hot issue to discuss. In this paper, liver biopsy in 58 cases of chronic hepatitis B patients and quantitative analysis the HBV cccDNA and HBV DNA in the liver tissue and detection serum HBV DNA at the same time. To explore the dynamics of hepatitis B virus covalently closed circular DNA (cccDNA) and optimal duration of treatment after serum virology response.[Methods]Detected 58 cases of chronic hepatitis B patients from August of 2007 to June of 2008 in Liver Diseases Apartment of Provincial Hospital affiliated to Shandong University. Male 48 cases and female 10 cases, ages from 19 to 66, average 34.8 years old. Among them, liver puncture before the anti-virus treatment were 23 cases and during the nucleoside analogue antiviral treatment were 35 cases. The diagnosis is consistent with "Guide to Combat Chronic Hepatitis B" and exclusion of hepatitis A, C, D, E and other reasons which may cause the liver damage. None of the cases received antiviral therapy before liver puncture in uncontrolled group. All cases of the controlled group have reached the serum virology response and liver function returned to normal before liver puncture (HBV DNA in peripheral blood <1000 copies/ml).There are 30 cases of patients with HBeAg negative conversion took place. HBV DNA and cccDNA were detected in liver sample by Roche light-cycle FQ-PCR. Serum HBV DNA was detected by ELISA.[Results]1. The cccDNA in biopsy samples continued to decrease after serum virology responded. The patients were divided into group a,b and c according to serum HBV DNA. HBV DNA and cccDNA in biopsy samples were no significantly difference between group a and b (Nemenyi display:χ~2= 0.1364, 0.0767, P = 0.7119, 0.7818); difference between group a and group c was significant (Nemenyi display:χ~2 = 20.5024,9.6948, P<0.0001, P = 0.0018); difference between group b and group c was significant (Nemenyi display:χ~2= 16.4618, 9.2824, P <0.0001, P = 0.0023), see Table 1.2. The longer last time after serum virology response, cccDNA in liver tissue more lower(r =- 0.370, P = 0.0142). 35 cases of patients were divided into group a,b and c according to the treatment time after serum virology response. HBV DNA levels in liver tissues were no significantly difference among the three groups (Kruskal-Wallis test: H = 4.9551, P = 0.0839). CccDNA levels in liver tissues were significantly difference between group a and b, c (Nemenyi display:χ~2= 6.4674, 16.4592, P = 0.0110, P <0.0001); the difference was also significantly between group b and c (Nemenyi display:χ~2 = 6.7682, P = 0.0093), see table 2.3. Anti-HBe positive patients had lower cccDNA in liver tissue than anti-HBe negative patients. 35 cases patients divided into group a,b and c according to HBeAg and anti-HBe. HBV DNA level in liver tissue was significantly difference between group b and c (Nemenyi display:χ~2= 7.3569, P = 0.0067). CccDNA levels in liver tissues were significantly difference among three groups. The continue treatment times were significantly difference among three groups, see Table 3. 4. There were 12 cases whose cccDNA in liver tissue undetected in 18 case HBeAb (+)patients. Serum virology response lasted 35 months and HBeAb (+) lasted 30 months.[Conclusion]1. HBV DNA, HBV cccDNA level in liver tissue had no significantly difference between sera HBV DNA≥10~7 copies / mL group and 10~3-10~6 copies / mL group.2. HBV DNA, HBV cccDNA level in liver tissue had significantly difference HBV between sera HBV DNA positive group and negative group.3. The longer the treatment time after serum virology response, the lower the cccDNA level in liver tissue.4. Anti-HBe positive patients had lower cccDNA in liver tissue than anti-HBe negative patients.5. There were 12 cases whose cccDNA in liver tissue undetected in 18 case HBeAb (+) patients. Serum virology response lasted 35 months and HBeAb (+) lasted 30 months.