大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植的实验研究

Objective To investigate a novel method of adrenocotical cells transplantation. Methods Adrenal glands of neonate rats were dissociated into adrenal cortical cells. Cells were cultured in self-made collagen type Ⅰ gel for one week, and then transplanted under the renal capsule of bilateral adrenalectomy mate rats. Blood samples were collected per week after surgery. Animals were sacrificed at the 8th week, and histological characteristics of the allografts, proliferation of transplanted cells, CYP11B1 and CYP11B2 expression as well as plasma aldosterone and corticosterone were observed. Results There were totally 15 rats receiving collagen gel transplantation, and 12 survived 8 weeks post-operation. The plasma cortocosterone level in collagen gel transplantation group was significantly higher than in adrenocortical cell transplantation group, and reached the normal level from 6th to 8th week, but the change in plasma aldosterone level in collagen gel transplantation group was similar to that of adrenocortical cell transplantation group. The adrenocortical cells cultured in gel grew well, and had a high proliferation rate. 95% of them were fasciculata cells which expressed CYP11B1, and the rest were glomerulosa cells which expressed CYP11B2. No inflammatory cell infiltration was observed. Conclusion The proliferation and function of adrenocortical cells could be promoted when they were cultured in collagen gel.     

以α6-integrin和c-kit为特异性表面标志分离筛选小鼠精原干细胞的可行性

背景:从目前文献报道来看,精原干细胞分离效率较高且较为公认的方法是通过隐睾模型结合表面标志进行多参数筛选.目的:探讨06-integrin和c-kil作为分离筛选精原干细胞特异性表面标志的可行性.设计、时间及地点:随机对照动物实验,于2006-05/12在武汉大学人民医院完成.材料:6周龄雄性昆明白小鼠40只,随机分为隐睾组、正常对照组,20只/组.方法:隐睾组小鼠建立隐睾模型,麻醉后腹部正中切口,将两侧睾丸拉入腹腔,将脂肪垫靠近附睾处各缝一针,分别固定在侧腹壁.正常对照组小鼠不进行任何干预.造模后两三个月,采用传统的两步消化法获取生精上皮单细胞悬液,加入FITC标记的抗α6-integrin抗体和PE标记的抗c-kit抗体,利用流式细胞仪进行α6-integrin+和c-kit双重筛选,锥虫蓝染色监测细胞活性.主要观察指标:隐睾的形态学变化,精原干细胞分选结果.结果:隐睾小鼠的生精小管内细胞排列紊乱,层次及管腔消失,小管中央可见分裂相细胞,细胞数明显减少.与正常对照组比较,隐睾组side scatter(low)、Forward scatter(hi)区的睾丸细胞增多,图形向左上移位.α6-integrin+和c-kit-细胞分布存在明显的相互偏离,即α6-integrin+细胞绝大多数不是精原干细胞,c-kit-细胞绝大多数也不是精原干细胞.从隐睾组筛选出具有sidescatter(low)、α6-integrin+、c-kit-特征的细胞数占睾丸细胞总数的2.8%,此即为精原干细胞,锥虫蓝监测结果显示细胞活性达95%以上.结论:采用α6-integrin和c-kit这两种表面标志进行精原干细胞的筛选,尽管可以提高细胞悬液中的精原干细胞纯度,但均缺乏特异性.     

五倍子鞣酸的体外杀精和抑菌作用研究

目的:研究五倍子鞣酸的杀精和抑菌作用。方法:按照WHO标准方法,对20例人精液进行体外杀精试验,并观察了五倍子鞣酸对大肠杆菌、金黄色葡萄球菌的最低抑菌浓度(MIC)和最低杀菌浓度(MBC),并将其效果与壬苯醇醚-9(NR-9)作比较。结果:五倍子鞣酸与人精液作用20s最低有效杀精浓度为20mg.mL-1。鞣酸对大肠杆菌(3株)MIC为0.195～0.390mg.mL-1,MBC为0.390～0.780mg.mL-1;对金黄色葡萄球菌(2株)MIC为0.049～0.098mg.mL-1,MBC为0.195～0.390mg.mL-1。而在鞣酸MIC、MBL浓度范围内NP-9无抑菌杀菌作用。结论:五倍子鞣酸具有极强的精液蛋白凝固作用和很好的杀精抑菌效果,可望成为一种安全、有效的阴道杀精抑菌剂,值得进一步研究。     

大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植的实验研究

Objective To investigate a novel method of adrenocotical cells transplantation. Methods Adrenal glands of neonate rats were dissociated into adrenal cortical cells. Cells were cultured in self-made collagen type Ⅰ gel for one week, and then transplanted under the renal capsule of bilateral adrenalectomy mate rats. Blood samples were collected per week after surgery. Animals were sacrificed at the 8th week, and histological characteristics of the allografts, proliferation of transplanted cells, CYP11B1 and CYP11B2 expression as well as plasma aldosterone and corticosterone were observed. Results There were totally 15 rats receiving collagen gel transplantation, and 12 survived 8 weeks post-operation. The plasma cortocosterone level in collagen gel transplantation group was significantly higher than in adrenocortical cell transplantation group, and reached the normal level from 6th to 8th week, but the change in plasma aldosterone level in collagen gel transplantation group was similar to that of adrenocortical cell transplantation group. The adrenocortical cells cultured in gel grew well, and had a high proliferation rate. 95% of them were fasciculata cells which expressed CYP11B1, and the rest were glomerulosa cells which expressed CYP11B2. No inflammatory cell infiltration was observed. Conclusion The proliferation and function of adrenocortical cells could be promoted when they were cultured in collagen gel.     

建立动态管理模式 发挥奖助学金作用

武汉大学人民医院自2007年参与研究生培养机制改革,2007级研究生开始推行新的奖助体系,并探索建立了奖助金的动态管理模式,此举广泛地调动了广大研究生及导师的积极性,内在的激励作用凸显,起到了让奖助金真正“奖学”、“助学”的作用。     

血必净注射液在群蜂蜇伤致多器官功能障碍综合征治疗中的应用

目的：探讨血必净注射液（中药复方制剂）在群蜂蜇伤所致多器官功能障碍综合征（MODS）治疗中的应用价值.方法：回顾分析我院住院治疗的60例群蜂蜇伤所致MODS患者临床资料,分为血必净组（31例）和对照组（29例）.对照组患者治疗措施包括局部处理、糖皮质激素的应用、脏器功能保护、血液净化治疗;血必净组则在上述治疗上加用血必净注射液治疗.比较2组患者治愈率、病死率和平均住院时间.结果：血必净组治愈率显著高于对照组（83.9％ vs 58.6％,P＜0.05）,病死率显著低于对照组（9.7％ vs 31.0％,均P＜0.05）,血必净组患者平均住院时间短于对照组[（11.2±3.4）d vs （13.6±4.6）d,P＜0.05].结论：群蜂蜇伤致MODS患者在传统综合治疗的基础上加用血必净注射液在缩短病程和改善预后方面具有一定作用.     

蓝舌病毒湖北株对小鼠前列腺癌RM-1细胞的杀伤效应

目的:体外研究蓝舌病毒湖北株3(BTV-HbC3)对小鼠前列腺癌细胞RM-1的感染性并探讨BTV-HbC3靶向性溶瘤的机制。方法:观察RM-1细胞感染BTV-HbC3的细胞病变效应;MTT法研究病毒致细胞病变率的特征;透射电镜观察感染病毒后细胞超微结构的变化;DNA Ladder分析病毒诱导细胞凋亡的情况;流式细胞仪测定病毒对RM-1细胞凋亡的影响。结果:BTV-HbC3感染RM-1细胞后有明显的细胞病变效应;DNA Ladder分析为阶梯状条带;透射电镜发现胞质内有大量病毒颗粒和典型细胞凋亡形态变化;流式细胞仪可见明显的细胞凋亡。结论:BTV-HbC3在体外能有效的感染RM-1细胞,并能诱导RM-1细胞凋亡。     

小鼠隐睾模型研究进展

隐睾症是指睾丸下降异常,使睾丸不能降至阴囊而停留在腹膜后、腹股沟管或阴囊入口处.隐睾的发病率为1.6% ～ 9.0%,先天隐睾患儿发病率则为0.9% ～ 1.8%,近年来的研究发现隐睾的患病率呈增高的趋势[1].     

增塑剂邻苯二甲酸二丁酯致小鼠睾丸组织的病理学改变

背景：目前国内外研究邻苯二甲酸二丁酯对生殖损害研究对象多为大鼠,且不同时间点下小鼠病理组织学未见,以小鼠为移植对象没有明确移植时间。 目的：探讨邻苯二甲酸二丁酯致小鼠睾丸组织病理学改变,并找出其改变最大的时间点,为移植做准备。 方法：妊娠balb/c小鼠20只随机分成3组,分别为正常对照组6只,玉米油对照组6只,DBP组8只。自妊娠12~21 d,分别经口给予邻苯二甲酸二丁酯和玉米油,分别在小鼠出生后4~8周每间隔1周观察仔代雄小鼠睾丸的组织病理学改变,找到变化最大的时间点。 结果与结论：邻苯二甲酸二丁酯组染毒小鼠睾丸出现明显的生理、病理和电镜下的改变。邻苯二甲酸二丁酯可引起雄性仔鼠性分化异常,睾丸生精上皮损害和生精过程障碍,从而对雄性仔鼠生育力产生不利影响,在5,6周小鼠睾丸组织损害最大,可以作为移植变化最大时间进行选择。