乙型肝炎病毒DNA經(jīng)精細(xì)胞垂直傳播的實(shí)驗(yàn)研究.pdf

1、Objective To detect hepatitis B virus (HBV) DNA in testis and epididymis of human with positive HBsAg and HBeAg, observe the capacity of sperm to capture HBV DNA, ascertain the position of HBV DNA in sperm cell, and es

2、tablish murine model on HBV DNA vertical transmission. Materials and Methods ①The testis and epididymis tissues of 22 men with HBV and 18 normal samples were collected. After HBV DNA was detected from the tissues by n

3、est PCR, Fluorescence in situ hybridization (FISH) was used to make certain the localization of HBV DNA. ②Murine sperm cells were cultured in vitro and were transfected HBV DNA. Then genome DNA of sperm cells were ext

4、racted and amplified by PCR, and the exact position of HBV DNA in sperm cells was observed by FISH. ③Establish murine model on HBV DNA vertical transmission using sperm as carriers and then observe the expression of H

5、BV DNA in the new generations by PCR and RT-PCR.Results ①HBV DNA were detected in the testis and epididymis tissues of 18 men with positive HBsAg and HBeAg, and the positive ratio was 81.8％. Relatively, HBV DNA was not

6、detected in normal samples. And so, there were significant differences between two groups (P＜0.001). HBV DNA mostly located in spermatogenic epithelium and Sertoli cell of testis and epididymis tissues by FISH. ②Under fl

7、uorescence microscope,HBV DNA was discovered in head and rear end of some germ cells, they were Kelly fluorescence granule or brown granule after DAB stained. ③There were 31 new generations's mice, eleven mice had HBV DN

8、A, the positive ratio was 35.4％(11/31).Among the eleven mice, only three had HBV RNA.Conclusion ①HBV DNA were detected in testis and epididymis tissues of human with HBV and located in all levels of germ cells. ②The ger

9、m cell can capture HBV DNA actively, so germ cell had the ability of carrying HBV DNA. ③Murine model on HBV DNA vertical transmission was established using sperm as carrier. HBV DNA could be transmitted from old generati