Objective To study a simple and practical method of isolation, culture and identification of hepatic oval cells from adult rat. Methods Wistar adult rats were fed by 2-acetaminofluorere （AAF） and were stimulated by partial hepatectomy to activate the proliferation of hepatic oval cells. After operation 12 days, the livers were resected for isolating oval cells. Hepatic tissue was digested by 0.10% collagenase Ⅳ and the obtained heterogeneous liver cells were then isolated and purified by density gradient centrifugation. The expressions of albumin and CK19 mRNA in hepatic oval cells were analyzed by immuno-fluorescence and RT-PCR. Results The survival rate of the newly isolated oval cells was more than 90%. The hepatic stem cells were shown by immuno-fluorescence of stem cell’s antigen c-kit. The expressions of mRNA CK19 and albumin of the oval cell were also detected by PCR. The proliferation activity of the newly isolated oval cells was significantly high and they could be induced to differentiate into both hepatic and bile ductal cells by some growth factors. Conclusion The successful development of the simple and feasible isolation and purification procedure as well as the identification method for hepatic oval cells may provide a fundamental for further studies about bionomics of the hepatic stem cell and the relation between stem cells and hepatic carcinoma.
目的 探讨腹腔镜胆囊切除术（laparoscopic cholecystectomy，LC）中不同CO2气腹压力对患者肝功能、动脉血气及对颈、肩痛的影响。方法 选择行择期LC术患者120例，采用随机数字表法，按住院先后顺序对应入组，随机均分为气腹压力10mmHg （1 mm Hg=0.133kPa）组、12mmHg组及14mmHg组3组；对比分析3组患者手术后肝功能、血气指标以及术后当天、第1和第2天恶心、呕吐及颈肩部疼痛的发生率。结果 3组患者术前各参数组间比较差异均无统计学意义（P＞0.05）。术后肝功能及血气分析指标的改变3组间的差异有统计学意义（P＜0.05），气腹压力越大，其术后AST、ALT及TBIL水平升高越明显，而pH及PO2的下降越明显，PCO2的升高也越明显；术后患者颈、肩痛及恶心呕吐发生率3组间比较差异也有统计学意义（P＜0.05），气腹压力越大，术后颈、肩痛及恶心呕吐发生率越高。结论 CO2气腹压力对术后肝功能及动脉血气指标的改变以及术后颈、肩痛和恶心呕吐发生率有明显影响。
目的总结困难腹腔镜胆囊切除术（LC）中造成操作困难的原因，探讨各种情况下的手术技巧。 方法回顾性分析2004年12月至2013年12月期间我院为128例LC术中遇有操作困难的患者施行手术的临床资料。 结果128例患者中113完成LC，住院时间为5～7 d（平均5.50 d）；15例中转开腹完成，住院时间为7～9 d（平均7.46 d）。术后发生胆汁漏1例，经保守治疗治愈，余术中、术后均未发生严重并发症。128例患者术后随访3个月～2年（平均19.4个月），无并发症发生。 结论术中仔细解剖，谨慎操作，采用适当的手术技巧，多数困难LC能顺利完成，遇复杂情况果断中转开腹是避免LC严重并发症的关键措施。