目的 :研究幽门螺杆菌空泡毒素作为单一毒力决定簇对真核细胞的作用。方法 :用 PCR扩增vac A基因片段 ,克隆入真核表达载体 p EGFP- N 1,构建重组质粒 p EGFP- vac A,转染 He L a细胞 ,通过相差显微镜和电子显微镜观察细胞形态与结构的...目的 :研究幽门螺杆菌空泡毒素作为单一毒力决定簇对真核细胞的作用。方法 :用 PCR扩增vac A基因片段 ,克隆入真核表达载体 p EGFP- N 1,构建重组质粒 p EGFP- vac A,转染 He L a细胞 ,通过相差显微镜和电子显微镜观察细胞形态与结构的变化。结果 :重组质粒转染 He L a细胞 2 4 h,10 %～ 2 0 %细胞的胞质内出现明显空泡 ,其中少数细胞发生凋亡改变。结论 :成功构建了用于真核表达的重组 vac A质粒 ,转染真核细胞后 ,观察 Vac A作用导致的细胞形态结构的变化 ,为研究 Vac展开更多
Objective: Since little is known about the actual incidence and fate of vacuoles at different stages of development this preliminary study was set up to accurately measure vacuoles and track them to day 5. Design: Pro...Objective: Since little is known about the actual incidence and fate of vacuoles at different stages of development this preliminary study was set up to accurately measure vacuoles and track them to day 5. Design: Prospective study. Setting: Women’s General Hospital in Austria. Patient(s): A total of 223 consecutive IVF and intracytoplasmic sperm injection (ICSI) cycles (206 patients). Intervention(s): Accurate measurement of vacuoles. Affected gametes and embryos were cultured individually and tracked until day 5. Main Outcome Measure(s): Size and number of vacuoles, fertilization rate, blastocyst formation rate, blastocyst quality. Result(s): There was a significant relationship between size of the vacuole (cutoff value 14 μ m) and fertilization (P< .05). At zygote stage the incidence of vacuoles was higher (P< .01) in ICSI (11.6% ) than in IVF (5.3% ). Only 32.2% of affected ICSI- embryos reached blastocyst stage on day 5 compared with 53.0% of the normal ones (P< .001). In terms of blastocyst formation vacuolization on day 4 (P< .001) turned out to be the most severe one. At blastocyst stage inner cell mass was affected less frequently than the trophectoderm (P< .05). Conclusion(s): Three types of vacuoles could be identified: (1) those already present at oocyte collection, which develop during maturation (day 0); (2) those artificially created by ICSI (day 1); and (3) those accompanied with developmental arrest (day 4). The later that vacuoles arose, the more detrimental their effect on blastocyst formation.展开更多
文摘Objective: Since little is known about the actual incidence and fate of vacuoles at different stages of development this preliminary study was set up to accurately measure vacuoles and track them to day 5. Design: Prospective study. Setting: Women’s General Hospital in Austria. Patient(s): A total of 223 consecutive IVF and intracytoplasmic sperm injection (ICSI) cycles (206 patients). Intervention(s): Accurate measurement of vacuoles. Affected gametes and embryos were cultured individually and tracked until day 5. Main Outcome Measure(s): Size and number of vacuoles, fertilization rate, blastocyst formation rate, blastocyst quality. Result(s): There was a significant relationship between size of the vacuole (cutoff value 14 μ m) and fertilization (P< .05). At zygote stage the incidence of vacuoles was higher (P< .01) in ICSI (11.6% ) than in IVF (5.3% ). Only 32.2% of affected ICSI- embryos reached blastocyst stage on day 5 compared with 53.0% of the normal ones (P< .001). In terms of blastocyst formation vacuolization on day 4 (P< .001) turned out to be the most severe one. At blastocyst stage inner cell mass was affected less frequently than the trophectoderm (P< .05). Conclusion(s): Three types of vacuoles could be identified: (1) those already present at oocyte collection, which develop during maturation (day 0); (2) those artificially created by ICSI (day 1); and (3) those accompanied with developmental arrest (day 4). The later that vacuoles arose, the more detrimental their effect on blastocyst formation.
