HPV核酸原位杂交技术的应用体会

尖锐湿疣是与人类乳头瘤病毒(HPV)感染有关的性传播疾病,据报道女性生殖道尖锐湿疣HPV6/¨的阳性表达占96.4%[1].仅凭H-E切片形态难以进行判断,HPV免疫组化阳性率很低,特异性不强,核酸原位杂交具有较高的敏感性和特异性.我室于1995年开始用原位杂交的方法鉴定HPV以确诊尖锐湿疣,反复实验对地高辛标记的探针在应用技术有了新的认识和体会.

Objective To study the biological effects of basic fibroblast growth factor (bFGF) in the process of embryo implantation in mouse Materials & Methods The transcription and translocation of bFGF and its receptor (Flg) in endometrium of 60 Kunming mice were detected with the methods of im munohistochemistry and hybridization in situ. Endometrial tissue was obtained on the day 4 from pregnancy mice. The cells were cultured and attached in 1 : 1 F12/DMEM (vol/vol) supplemented with 10% FBS for 24 h The medium supplemented with 1% FBS and bFGF (50 ng/mL), anti-bFGF antibody (250 ng/mL) or anti-Flg antibody (250 ng/mL) was added in different culture wells. At different culture stages, the biological effect of bFGF on cell survival and expression of LN, FN and c-myc was detected by using MTT analysis, immunohistochemistry and hybridization in situ.Results bFGF and Flg was located in luminal and glandual cells on day 4 and 5 of pregnancy. Embryonic implantation was accompanied by increased bFGF and its recep tor in decidual cell around implantation site, in which low level of bFGF and its recep tor was apparent on day 7 and 8 of pregnancy. In vitro, the OD value in culture wells containing bFGF was significantly higher than that in control group. Exogenous bFGF promoted the expression of LN, FN and c-myc.Conclusion Changes in the cell-specific distribution of bFGF and the effects of bFGF on cultured endometrial cells imply a multifunctional role of the growth factor in uterine cell proliferation, differentiation and embryonic implantation.