The method of laser capture microdissection (LCM) combined with suppressive subtractive hybridization (SSH) was developed to isolate specific germ cells from human testis sections and to identify the genes expressed d...The method of laser capture microdissection (LCM) combined with suppressive subtractive hybridization (SSH) was developed to isolate specific germ cells from human testis sections and to identify the genes expressed during differentiation and development. In the present study, over 10,000 primary spermatocytes and round spermatid cells were successfully isolated by LCM. Using the cDNAs from primary spermatocytes and round spermatids, SSH cDNAs library of primary spermatocyte-specific was constructed. The average insert size of the cDNA isolated from 75 randomly picked white clones was 500 bp, ranging from 250 bp to 1.7 kb. Using the dot-blot method, a total of 421 clones were examined, resulting in the identification of 390 positive clones emitting strong signals. Partial sequence of cDNAs prepared from each clone was determined with an overall success rate of 84.4%. Genes encoding cytochrome c oxidase II and the rescue factor-humanin were most frequently expressed in primary spermatocytes, suggesting their roles involved in meiosis.展开更多
目的研究深圳城区育龄妇女与高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)感染率和宫颈糜烂患病率的年龄分布特点及相关性。方法 2004年3月～2008年3月期间对深圳市城区育龄妇女1943例进行宫颈癌筛查,包括问卷调查、妇...目的研究深圳城区育龄妇女与高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)感染率和宫颈糜烂患病率的年龄分布特点及相关性。方法 2004年3月～2008年3月期间对深圳市城区育龄妇女1943例进行宫颈癌筛查,包括问卷调查、妇科检查,收集宫颈脱落细胞。采用第二代杂交捕获(HC-2)法检测13种高危型HPV。结果深圳城区妇女HR-HPV DNA阳性381例,阳性率19.6%。各年龄组感染率之间有显著性差异(χ2=28.67,P<0.01)。宫颈糜烂患病率为41.8%,从年龄分布来看,不同年龄组间宫颈糜烂患病率有显著性差异(χ2=81.08,P<0.001)。其中患病率最高的是25～29岁组55.2%。在各年龄组轻度及中度宫颈糜烂患病率差异无显著性(P>0.05)。但在重度宫颈糜烂百分比中25～29岁组重度糜烂患病率差异有显著性(P<0.05)。宫颈光滑、宫颈糜烂患者的HR-HPV感染率分别是15.8%及24.8%,宫颈糜烂患者感染HR-HPV的OR值为1.569,95%CI(1.310～1.878)。结论①深圳城区妇女生殖道HR-HPV感染的年龄分布存在着两个感染高峰年龄段(25～29岁及55～60岁),宫颈糜烂患病率存在一个患病高峰年龄段(25～29岁);②25～29岁组是患重度宫颈糜烂的高发年龄段;③宫颈糜烂是年轻女性感染生殖道HR-HPV的高危因素。展开更多
文摘The method of laser capture microdissection (LCM) combined with suppressive subtractive hybridization (SSH) was developed to isolate specific germ cells from human testis sections and to identify the genes expressed during differentiation and development. In the present study, over 10,000 primary spermatocytes and round spermatid cells were successfully isolated by LCM. Using the cDNAs from primary spermatocytes and round spermatids, SSH cDNAs library of primary spermatocyte-specific was constructed. The average insert size of the cDNA isolated from 75 randomly picked white clones was 500 bp, ranging from 250 bp to 1.7 kb. Using the dot-blot method, a total of 421 clones were examined, resulting in the identification of 390 positive clones emitting strong signals. Partial sequence of cDNAs prepared from each clone was determined with an overall success rate of 84.4%. Genes encoding cytochrome c oxidase II and the rescue factor-humanin were most frequently expressed in primary spermatocytes, suggesting their roles involved in meiosis.
