- 现金
- 3700 元
- 精华
- 16
- 帖子
- 1790
- 注册时间
- 2002-12-9
- 最后登录
- 2021-4-14
|
3楼
发表于 2004-8-26 22:54
Patients and Methods
Patients
Between June 1998 and December 2001, 94 consecutive adult patients from San Giovanni Battista Hospital (Turin, Italy) with e-CHB were treated with Lam 100 mg dose daily. They were followed up until 31 January 2003 and were included in this retrospective analysis. Median duration of treatment was 33 months, between 12 and 54.
The demographic and clinical features of patients are outlined in Table 1.
Criteria for inclusion in the analysis were: (i) HBsAg-positive for at least 6 months, HBeAg-negative and anti-HBe-positive; (ii) serum alanine aminotransferase (ALT) values >1.5 times the upper limit of normal (ULN) in at least two determinations during the 6 months preceding therapy; (iii) positive serum HBV DNA, tested by hybridization assay, non-amplified technique (>10E5 copies/mL); (iv) CH or compensated liver cirrhosis. We did not include patients co-infected by the HCV, HDV and HIV.
Chronic liver disease was confirmed by a liver biopsy taken within 18 months before therapy in 59 patients: 50 had chronic active hepatitis, nine had cirrhosis. Among the 35 patients for whom no liver biopsy was available, 13 were considered to have cirrhosis (nine patients Child-Pugh A and four Child-Pugh B grade), diagnosed on the basis of signs of portal hypertension by upper endoscopy (six patients had oesophagus varices) or ultrasound. The remaining 22 patients had persistently abnormal ALT levels without clinical signs of cirrhosis. They refused liver biopsy before therapy.
Thirty-one patients had received prior treatment with α-interferon (IFN): eight had relapsed during therapy after an initial response and 23 had not responded. Three patients had received prior treatment with Famciclovir and one with Entecavir. All previous treatments were discontinued at least 6 months before Lam therapy. None patient was previously treated with Lam.
Methods
Follow-Up. The patients were followed-up at 3-month intervals. Before therapy and at each clinic visit routine haematological, biochemical and virological tests [HBsAg, HBsAb, HBeAg, HBeAb, IgM-antiHBc and HBV DNA by polymerase chain reaction (PCR)] were performed. The patients who had a virological breakthrough (VBK) and those who did not respond to therapy were tested for HBV polymerase mutants by Inno-Lipa HBV DR assay. Seventeen patients with different therapy outcomes were analysed retrospectively for sequence variations in the core-promoter, precore and polymerase rt-domain region B/C at baseline, and during therapy (at the appearance of VBK and at the end of follow-up).
All patients underwent ultrasound of the upper abdomen every 6 months.
Paired liver biopsies taken at baseline and during Lam therapy were obtained in 20 patients.
Therapy Outcome Definitions
The outcome was defined as (i) virological response (VR): negative HBV DNA by PCR for at least two consecutive determinations, (ii) biochemical response (BR): decrease in serum ALT to within the normal range during therapy, (iii) VBK:[8] HBV DNA-positive by PCR (>1000 copies/mL) for at least two consecutive determinations after an initial VR, (iv) no response (NR): persistence of HBV DNA viraemia detectable by PCR on-therapy.
Serology. Serum HBV markers (HBsAg, HBsAb, HBeAg, HBeAb, IgM anti-HBcore), anti-HCV, anti-HDV and anti-HIV were tested with immunoenzymatic assays (EIA; Abbott Diagnostic Inc., Chicago, IL, USA). HBV DNA load was performed by Dygene Hybrid-Capture I (sensitivity 700 000 copies/mL) at baseline, and during therapy by an amplified technique, PCR (Amplicor HBV, Monitor; Roche, Branchburg, NJ, USA; sensitivity of 1000 copies/mL).[17]
HBV Polymerase Mutants Assay. Polymerase mutants were determined using a commercially available kit (Inno-LiPa HBV DR, Innogenetics N.V., Belgium) based on reverse hybridization of PCR amplified HBV DNA against specific probes immobilized on paper strips.[18,19]
Genotyping. The HBV genotype was determined using a PCR-based method with primers specific for the six major genotypes (A-F).[20]
Statistical Analysis
Patients who were lost at follow-up were considered as failure of treatment. All the data were analysed by stats direct software, version 1.21. The Kaplan-Meier' method[21] was used to estimate the cumulative rate of biochemical and virological remission during the study period. Statistical significance analysis of the histological response was performed with the Wilcoxon rank sum test. A P-value of <0.05 was considered statistically significant. |
|