H1foo is essential for in vitro meiotic maturation of bovine oocytes.

Oocyte-specific linker histone, H1foo, is localized on the oocyte chromosomes during the process of meiotic maturation, and is essential for mouse oocyte maturation. Bovine H1foo has been identified, and its expression profile throughout oocyte maturation and early embryo development has been established. However, it has not been confirmed if H1foo is indispensable during bovine oocyte maturation. Effective siRNAs against H1foo were screened in HeLa cells, and then siRNA was microinjected into bovine oocytes to down-regulate H1foo expression. H1foo overexpression was achieved via mRNA injection. Reverse transcription polymerase chain reaction (RT-PCR) results indicated that H1foo was up-regulated by 200% and down-regulated by 70%. Based on the first polar body extrusion (PB1E) rate, H1foo overexpression apparently promoted meiotic progression. The knockdown of H1foo significantly impaired bovine oocyte maturation compared with H1foo overexpression and control groups (H1foo overexpression = 88.7%, H1foo siRNA = 41.2%, control = 71.2%; P < 0.05). This decrease can be rescued by co-injection of a modified H1foo mRNA that has escaped from the siRNA target. However, the H1e (somatic linker histone) overexpression had no effect on PB1E rate when compared with the control group. Therefore we concluded that H1foo is essential for bovine oocyte maturation and its overexpression stimulates the process.

[The relationship between HBV genotypes and the efficacy of antiviral therapies in hepatitis B e antigen- positive patients].

OBJECTIVES: To study the relationship between HBV genotypes and the efficacy of antiviral therapies. METHODS: HBV genotypes of 90 hepatitis B e antigen positive patients with chronic hepatitis B (CHB) were determined by PCR sandwich hybridization-ELISA technique. Forty-one patients with CHB were treated with lamivudine (100 mg/day) for 48 weeks and 49 patients with CHB were given alpha-interferon (3 MU/QOD) therapy for 48 weeks. The serological, biochemical and virological symbols were measured before, during and after treatment for all the patients. RESULTS: Of the 90 patients, genotype B HBV was found in 16 and C in 74. There was no difference in the rate of response to lamivudine treatment between patients with genotype B or C HBV (33.3% vs. 20.0%) after 48 weeks treatment with lamivudine in the 41 patients. Of the 49 HBeAg positive CHB patients treated with alpha-interferon for 48 weeks, in HBV genotype B and C patients the rates of normalization of ALT were 60.0% and 20.5%; the rate of HBeAg turning to negativity was 50.0% and 17.9%; and the rate of HBV DNA undetectability was 50.0% and 17.9%. The rate of response to the interferon treatment was significantly higher in patients with HBV genotype B compared to those with genotype C. CONCLUSIONS: Our study shows that there is no influence on the lamivudine treatment effects for the HBV genotype B and C CHB patients, but the alpha-interferon treatment for HBV genotype B CHB patients is more effective than that for the genotype C ones.

[Liver pathological changes and clinical features of patients with chronic hepatitis B virus infection in their immune tolerant phase and non-active status].

OBJECTIVE: To investigate the liver pathological changes and the clinical features of patients with hepatitis B virus (HBV) infection in their immune tolerant phase and non-active status. METHODS: Fifty-four patients with chronic HBV infection in their immune tolerant stage and another 47 patients with the same infection but in non-active status were involved in this study. Statistical analysis including the ages and sex of the patients, their serum levels of HBV DNA, hepatocytic expression of HBsAg and HBcAg and their liver pathology were studied and statistically analyzed. Histological grading of inflammation and staging of fibrosis in the livers were also compared and analysed in patients with different levels of serum ALT. RESULTS: The sex ratio of the two groups was of no significant difference. The average age of the patients in the non-active status [(28.11+/-8.60) years.] was older than that of the patients in the immune tolerant stage [(24.93+/-7.21) years], showing a significant difference (P < 0.05). The serum levels of HBV DNA of the patients in the immune tolerant stage were high and 94% of them had a HBV DNA higher than 106 copies/ml. In the non-active status group, 89% of the patients were HBV DNA negative. Between the two groups of patients there were no significant differences in the histological grades of liver inflammation or in the hepatocytic expressions of HBsAg and HBcAg. The stage of fibrosis was higher in the non-active status group than in the immune tolerant stage group, showing a significant difference between these two groups (u = 2.004, P < 0.05). The fibrosis stages of the livers of patients of a higher but within normal ALT level were markedly higher than those of a lower but within normal ALT level patients (u = 3.274, P less than 0.01). CONCLUSION: Patients infected with HBV in non-active status may have experienced some occult courses of immune active stages; they are older in age and have higher levels of fibrosis. ALT sustained at a high level but within the normal range may indicate a higher degree of fibrosis, therefore liver pathological studies should be recommended for this kind of patient.

[Relationship between the quantities of peripheral dendritic cells and of serum HBV DNA and the inflammatory reaction levels in the liver].

OBJECTIVES: To investigate the relationship between the quantity of peripheral dendritic cell (DC) and of serum HBV DNA and the inflammatory level in chronic hepatitis B (CHB). METHODS: The myeloid DC (DC1) and plasmacytoid DC (DC2) in fresh peripheral blood were enumerated by using three-color flow cytometry in chronic hepatitis B patients and healthy donors. The hepatic inflammatory levels were evaluated by percutaneous liver biopsy. The serum HBV DNA levels were determined by real-time PCR. RESULTS: CHB patients with serum HBV DNA < or = 10(6) copies/ml exhibited a significant increase in the percentage of circulating DC2 in comparison with those of CHB patients with serum HBV DNA > or = 10(6) and with healthy donors (P < 0.05). The two latter groups showed no significant differences between each other. There was also no significant difference in the relative quantity of peripheral blood DC1 among the three groups mentioned above (P = 0.162). No evidence was found to support that the relative quantity of peripheral blood DC2 was associated with the clinical severity of the disease or the inflammatory level in the liver (P > 0.05). CONCLUSION: The relative quantity of peripheral blood DC2 is associated with HBV DNA level. It is suggested that DC2 may play a pivotal role in inhibiting HBV replication in CHB patients. There was no relationship found between relative quantities of DCs and the inflammatory level in the liver.