目的应用蛋白质组学实验方法对在异体异位生长发育的新生小鼠睾丸组织进行差异蛋白分析,从而对睾丸组织体外移植模型用于生精细胞发育研究的可行性提供进一步的理论依据。方法以新生小鼠睾丸组织为供体,免疫缺陷小鼠为受体进行组织移植...目的应用蛋白质组学实验方法对在异体异位生长发育的新生小鼠睾丸组织进行差异蛋白分析,从而对睾丸组织体外移植模型用于生精细胞发育研究的可行性提供进一步的理论依据。方法以新生小鼠睾丸组织为供体,免疫缺陷小鼠为受体进行组织移植,移植8周后收集移植物组织,采用双向凝胶电泳(2-DE)分离移植物组织总蛋白,同时分离8周正常小鼠睾丸组织总蛋白作对比分析,运用Im ageM aster 2D E lite 5.0图像分析软件识别移植物组织与正常组织差异表达的蛋白质点,应用基质辅助电离解析飞行时间质谱(MALD I-TOF-MS)获取肽质量指纹图谱(PMF),检索数据库鉴定差异表达的蛋白质点。结果正常睾丸组织和移植物组织双向电泳图谱的平均蛋白质点数分别为(1 326±15)个和(1 561±20)个,3次重复实验的蛋白质点位置重复性较好。通过比较两者的双向凝胶电泳图谱,得到表达差异量较大的蛋白质点21个。选取6个仅在正常睾丸组织中有表达的蛋白质点进行质谱和生物信息学分析,鉴定出的6种蛋白质,除血红蛋白α1链(Hbα-α1)外,其他5种蛋白均为在小鼠睾丸中有较高表达的蛋白质,包括A激酶锚定蛋白结合的精子蛋白(ASP)、磷脂过氧化氢物谷胱甘肽过氧化物酶(GPX4)、精子蛋白(Sp17)、天门冬酰胺酶样蛋白(ASRGL1)和过氧化物氧化还原因子4(Prdx4)。结论在本实验中,从异体异位生长发育成熟的新生小鼠睾丸组织中检测到5种在睾丸中特异性高表达蛋白质的缺失,这些蛋白质主要在精子运动、精子顶体形成以及精子的受精等功能中发挥作用。展开更多
To investigate ovarian function after heterotopic autotransplantation of human ovarian tissue banked at -196°C. A clinical case study. University medical center. A 37-year-old woman with cervical cancer (stage Ib...To investigate ovarian function after heterotopic autotransplantation of human ovarian tissue banked at -196°C. A clinical case study. University medical center. A 37-year-old woman with cervical cancer (stage Ib). Frozenthawed human ovarian tissue was transplanted to two different heterotopic sites. Ovarian function of the grafts was monitored sequentially by blood sampling for the hormonal profiles and by ultrasound. The hormonal profile remained at the postmenopausal level until 10 weeks after transplantation. By 14 weeks, the return of ovarian function was evidenced by the elevation of the serum E2 level (57.5 pg/mL). While monitoring hormonal profiles every 2 days for 5 weeks, we observed the LH surge (69.8 IU/L) followed by the elevation of the P4 concentration (9.6 ng/mL), suggesting presumptive ovulation. The ultrasound revealed a dominant follicle on the rectus muscle in the abdominal site. However, there was no sign of follicle development in the breast site. Ovarian function ceased around 28 weeks after transplantation. Heterotopic autotransplantation of cryobanked human ovarian tissue can be a practical strategy for restoration of ovarian function after cancer treatment. As a site for transplantation, a space between the rectus sheath and the rectus muscle appeared to be effective.展开更多
文摘目的应用蛋白质组学实验方法对在异体异位生长发育的新生小鼠睾丸组织进行差异蛋白分析,从而对睾丸组织体外移植模型用于生精细胞发育研究的可行性提供进一步的理论依据。方法以新生小鼠睾丸组织为供体,免疫缺陷小鼠为受体进行组织移植,移植8周后收集移植物组织,采用双向凝胶电泳(2-DE)分离移植物组织总蛋白,同时分离8周正常小鼠睾丸组织总蛋白作对比分析,运用Im ageM aster 2D E lite 5.0图像分析软件识别移植物组织与正常组织差异表达的蛋白质点,应用基质辅助电离解析飞行时间质谱(MALD I-TOF-MS)获取肽质量指纹图谱(PMF),检索数据库鉴定差异表达的蛋白质点。结果正常睾丸组织和移植物组织双向电泳图谱的平均蛋白质点数分别为(1 326±15)个和(1 561±20)个,3次重复实验的蛋白质点位置重复性较好。通过比较两者的双向凝胶电泳图谱,得到表达差异量较大的蛋白质点21个。选取6个仅在正常睾丸组织中有表达的蛋白质点进行质谱和生物信息学分析,鉴定出的6种蛋白质,除血红蛋白α1链(Hbα-α1)外,其他5种蛋白均为在小鼠睾丸中有较高表达的蛋白质,包括A激酶锚定蛋白结合的精子蛋白(ASP)、磷脂过氧化氢物谷胱甘肽过氧化物酶(GPX4)、精子蛋白(Sp17)、天门冬酰胺酶样蛋白(ASRGL1)和过氧化物氧化还原因子4(Prdx4)。结论在本实验中,从异体异位生长发育成熟的新生小鼠睾丸组织中检测到5种在睾丸中特异性高表达蛋白质的缺失,这些蛋白质主要在精子运动、精子顶体形成以及精子的受精等功能中发挥作用。
文摘To investigate ovarian function after heterotopic autotransplantation of human ovarian tissue banked at -196°C. A clinical case study. University medical center. A 37-year-old woman with cervical cancer (stage Ib). Frozenthawed human ovarian tissue was transplanted to two different heterotopic sites. Ovarian function of the grafts was monitored sequentially by blood sampling for the hormonal profiles and by ultrasound. The hormonal profile remained at the postmenopausal level until 10 weeks after transplantation. By 14 weeks, the return of ovarian function was evidenced by the elevation of the serum E2 level (57.5 pg/mL). While monitoring hormonal profiles every 2 days for 5 weeks, we observed the LH surge (69.8 IU/L) followed by the elevation of the P4 concentration (9.6 ng/mL), suggesting presumptive ovulation. The ultrasound revealed a dominant follicle on the rectus muscle in the abdominal site. However, there was no sign of follicle development in the breast site. Ovarian function ceased around 28 weeks after transplantation. Heterotopic autotransplantation of cryobanked human ovarian tissue can be a practical strategy for restoration of ovarian function after cancer treatment. As a site for transplantation, a space between the rectus sheath and the rectus muscle appeared to be effective.
