| Despite the availability of an effective vaccine, infection with hepatitis B virus (HBV) is still one of the major world health problems. This is mainly due to the fact that more than 350 million people are chronically infected with HBV world wide, who are at high risk of developing liver cirrhosis and hepatocellular carcinoma (HCC), accounting for 1 million to 2 million deaths annually. Recently with the usage of hepatitis B vaccine, the risk of horizontal transmission has reduced. However, effective methods for prevention of hepatitis B intrauterine infection which isn't impeded by vaccines are still not available. Once the intrauterine infection is installed, the neonate will develop to chronic hepatitis probably and become a harmful carrier. At present, the molecular merchanism of HBV infection hasn't been clarified, and a breakthrough progress haven't been made in the treatment of CHB (chronic hepatitis B). Study on the mechanism of HBV infection will lay solid foundation for developing more effective vaccines and drugs.Most chronic hepatitis B which tend to develop to immunity tolerance and complicated diseases pattens including HBsAg carrier, chronic hepatitis B, cirrosis and carcinoma are developed from intrauterine infection in China. Now the research on host response in infection diseases are being paid more attention, espically the relationship between host and complicated diseases patten after HBV infection are becoming pop field. The latest studies have showed that the host-virus interactions which determine the happen, development and outcome of disease extend far beyond those occurring at the cell surface but probably is a complex each other functionary network system. These interactions perform variation of gene expression patterns at molecular level. To study on integrated gene expression patterns will be useful to clarify the molecular mechanism of HBV infection and find more anti-virus targets. As the developing of strategies and methods of identification of novel genes, various methods to study patterns of gene expression have been described. It is becoming possible to identify some novel genes related to HBV infection in hepatocytes. The public articles about differential expression genes patterns concerning HBV infection in hepatocytes using SSH technology have not been seen. The aim of our studies is to clone and identify differentially expression genes of HBV infection in human fetal hepatocytes, and make definite their structures and functions in order to abtain the key factors of HBV-host interactions and establish foundation for finding novel anti-virus targets and drugs.In the study described here, over 22-weeks-old human primary fetal hepatocytes were isolated and cultured, and inoculated with HBV Dane particles. HBsAg and HBeAg in supernatant were detected by ELISA, HBcAg in nuclei by immunohistochemistry, HBV DNA in supernatant and cells by quantitative fluorescence PCR, and cccDNA in cultured cells by nested PCR in order to determine that the hepatocytes have been infected by HBV. Meanwile we have compared the sensitivity and specificity of these assays to verify some sensive and specific methods to indentify HBV infection. According to the documents, fibronectin can acceletate the growth and proliferation of cultured cells, and fibronectin in liver sinus can facilitate hepatocytes to take in HBV particles. So the effect of fibronectin on the growing status of fetal hepatocytes and it's effect on HBV infection in hepatocytes was also observed. The suppression subtractive hybridization(SSH) was performed by utilizing fetal hepatocytes infected with HBV as tester and those without HBV infection as driver. The cDNA fragments generated by SSH were cloned into pCR 2.1 vector and the subtracted cDNA library was constructed. The dot blots hybridization was used to screen the subtracted cDNA library clone with tester, driver, forward-and reverse-subtracted cDNA probes. The differentially expressed cDNA fragments was sequenced and analyzed with Blasta. The major results ar...
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