[Oxidative stress in patients with chronic hepatitis B before and after interferon alpha-2b treatment].

OBJECTIVE: To investigate the impacts of interferon alpha-2b (IFN alpha-2b) on the oxidative stress states in the treatment of chronic hepatitis B (CHB) with different genotypes. METHODS: Thirty-five patients with chronic hepatitis B and 18 healthy volunteers as a control were enrolled in this present study. In control and patients group, the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), serum malondialdehyde (MDA) levels, serum total antioxidative stress capacity (TAC) were measured spectrophotometrically. After the therapy with interferon alpha-2b at the dose of 300 million units via intramuscular injection thrice a week for 12 weeks, these parameters were measured again in the patient group. The genotypes of hepatitis B virus were detected by polymerase chain reaction and hybridization. The effective group was defined as the patients with complete response and partial response. RESULTS: The elevated concentrations of MDA and impaired levels of TAC in the patients with CHB were observed as compared to the healthy controls (P < 0.05 for both). There were no significant differences in serum levels of MDA and TAC in CHB patients with various genotypes (P > 0.05). The serum levels of MDA after the treatment with IFN alpha-2b were significantly lower than the pretreatment levels (P < 0.05), which even returned to the normal concentration (P > 0.05) in the effective group. There were significant increases in the TAC after the IFN alpha-2b therapy in the effective group. However, the significant differences in the TAC levels before and after the INFalpha-2b treatment were not observed in the non-responsive group. CONCLUSION: The oxidative stress could be improved with IFN alpha-2b treatment of chronic hepatitis B patients. The results suggest that antioxidant treatment for chronic hepatitis B patients may help improve the effect of anti-virus therapy.

Establishment and application of a TaqMan real-time quantitative reverse transcription-polymerase chain reaction assay for rubella virus RNA.

The aim of this study was to establish and apply a real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) for rubella virus (RV) RNA. First, the primer and TaqMan probe concentrations, as well as reaction temperatures were optimized to establish an efficient real-time quantitative RT-PCR assay for RV RNA. Next, an RV-specific PCR amplicon was made as an external standard to estimate the linearity, amplification efficiency, analytical sensitivity and reproducibility of the real time quantitative assay. Finally, the assay was applied to quantify RV RNA in clinical samples for rubella diagnosis. The RV-specific PCR amplicon was prepared for evaluation of the assay at 503 bp, and its original concentration was 2.75x109 copies/mul. The real time quantitative assay was shown to have good linearity (R2=0.9920), high amplification efficiency (E=1.91), high sensitivity (275 copies/ml), and high reproducibility (variation coefficient range, from 1.25% to 3.58%). Compared with the gold standard, the specificity and sensitivity of the assay in clinical samples was 96.4% and 86.4%, respectively. Therefore, the established quantitative RT-PCR method is a simple, rapid, less-labored, quantitative, highly specific and sensitive assay for RV RNA.

[Study on prenatal diagnosis using fluorescence quantitative polymerase chain reaction for congenital toxoplasmosis].

OBJECTIVE: To investigate prenatal diagnosis and treatment of toxoplasmosis in fetuses with fluorescence quantitative polymerase chain reaction (FQ-PCR) technique. METHODS: Of the 70 pregnant women with toxoplasma (TOX) DNA positive, TOX DNA in amniotic fluid and/or fetal umbilical cord blood was detected with FQ-PCR technique to diagnose fetal infection. 48 ones were given routine treatment with spiramycin for 2 therapy periods. Ultrasound examination were undertaken in all of pregnant women to monitor fetal growth. RESULTS: Of the 70 cases with TOX DNA positive, TOX DNA was detected in 21 fetuses. TOX DNA positive rates were similar in amniotic fluid and umbilical cord blood. The higher the TOX DNA, the higher fetal infectious rate. Fetal infectious rate was lower in treatment group (21%) than that in control group (50%), there was a statistically difference between two groups. CONCLUSIONS: Maternal TOX infection may cause fetal damage. Detection of TOX-DNA in amniotic fluid with FQ-PCR technique can diagnose fetal toxoplasmosis exactly. Treatment in pregnant period may decrease intrauterine infection rate.