BrdU作为脂肪干细胞标记示踪法的可行性

目的:研究BrdU标记猪脂肪干细胞(adipose-derived stem cells,ADSCs)的最佳剂量及时间,探讨其作为干细胞标记示踪方法的可行性.方法:自中华实验猪背部提取脂肪组织,采用贴壁法分离培养ADSCs,取第3代细胞以终浓度分别为10、15、20、25及30 μmol/L BrdU进行标记:另一孔不含BrdU作为对照组,分别培养12、24、48、72及96 h.免疫荧光法检测各组细胞BrdU标记率,找出BrdU的最佳标记方法,通过台盼蓝排斥试验、MTT及细胞凋亡检测,观察BrdU对ADSCs生长情况的影响.对第3代的ADSCs采用最佳标记方法后更换普通培养基继续培养,适时传代,连续检测第4、5、6、7、8代ADSCs的BrdU标记率.结果:原代培养的ADSCs的形态主要为长梭形,第3代ADSCs经BrdU标记后胞核呈红色荧光,随浓度的升高及时间的延长,BrdU阳性标记率逐渐升高,以终浓度20μmol/L BrdU标记48 h后阳性率达90%以上,且连续传5代标记率仍达40%.MTT、台盼蓝排斥试验及细胞凋亡检测发现BrdU对ADSCs生长增殖基本无影响.结论:BrdU标记ADSCs的最佳剂量和时间为20 μmol/L和48 h,该方法标记率高,对细胞影响小,可用于动态研究ADSCs在移植体内生长、分化.     

慢加急性肝衰竭病因及其病理生理机制研究进展

慢加急性肝衰竭(acute-on-chronic liver failure,ACLF)是国内最常见的肝衰竭类型,其临床表现复杂,病死率极高.ACLF的病理生理学改变目前尚不明确,深入了解ACLF的病理生理学改变有助于临床治疗工作的改善.近年来ACLF的病理生理学研究取得了一定的成就.本文就ACLF的概念、病因及其病理生理学改变作一综述.     

妊娠期急性脂肪肝

妊娠期急性脂肪肝（AFLP）是一种少见的妊娠晚期并发症，常发生在孕35周以后，但也有孕23周发病的报道。该病发病急骤，病情凶险，母儿死亡率高，以黄疸、凝血障碍、肝性脑病和肝脏小脂肪滴脂肪变性为特征。近年来随着对该病认识的不断深入，早期诊断和早期治疗可使本病预后得到极大改善，但由于本病常合并多脏器功能障碍，死亡率仍较高，进一步认识其发病机制和临床表现可为临床诊治提供更好的理论基础。     

兔慢性肝损害急性肝衰竭动物模型的建立

Objective To establish an ideal animal model of acute-on-chronic liver failure (ACLF) in New Zealand white rabbits in order to provide a large animal model for further researches.Methods Totally 75 New Zealand rabbits were randomly divided into experimental group (n =70) and control group (n = 5 ). Rabbits in the experimental group were injected with CCl4 into the abdominal cavity twice every week and the doses of CCl4 were modified according to the index of liver function and the body weight, whereas those in the control group were treated with the same volume of saline. At the 10th week,48 New Zealand rabbits with hepatic fibrosis were randomly assigned to 4 groups and injected with CCl4 as before, D-Gal at a dose of 0. 65 g/kg body weight (BW), 0. 70 g/kg BW and 0. 75 g/kg BW, respectively. By observing and comparing the general state, survival time, biochemical indexes, and the histopathology, a method of establishing a stable animal model of acute hepatic failure was found. Results As compared with those in control group, the levels of ALT, AST, GGT, HA, LN and PC-Ⅲ in the experiment group were increased significantly, while the level of ALB was decreased at the end of 10 weeks. Typical features of hepatic fibrosis and the formation of pseudo-lobules were observed at the end of 10 weeks. After treatment with D-Gal, all rabbits in group Ⅰ survived with minimal changes in liver function tests. In group Ⅱ , there was a temporary hepatic injury, but no hepatic coma. Four of the 12 rabbits died (33. 3% ). In group Ⅲ , biochemical indexes changed obviously 12 h after the administration and hepatic injury reached its peak after 48 h. Ten of 12 rabbits were died of severe hepatic failure with a survival time of ( 53. 00 ± 25. 69) h. Histology of liver section revealed massive necrosis in nodules. In group Ⅳ , hepatic injury occurred early and severely. All the rabbits died of severe hepatic failure with a survival time of (32. 70 ± 17. 46) h. Conclusion The experimental model of ACLF could be established by injected with D-Gal in New Zealand rabbits with hepatic fibrosis, induced by CCl4 intraperitoneal injection for 10 weeks.The one induced by 0. 70 g/kg of D-galactosamine was more stable and showed similar clinical pathophysiological changes in human beings. So it can be a good experimental platform for studies of ACLF.     

miR-21通过调节PTEN及Wnt信号通路介导Huh7细胞对顺铂的耐药性

目的探讨microRNAs(miRNAs)介导肝细胞癌(HCC)对顺铂的耐药机制。方法采用miR-21 mimics及miR-21抑制剂来调节Huh7细胞miR-21表达量。人DKK-1与bpv(phen)分别是Wnt信号通路抑制剂及PTEN抑制剂。相关mRNA及蛋白水平采用RT-PCR及蛋白质免疫检测。结果 miR-21过表达将减弱顺铂(5和10μg/mL)对Huh7细胞增殖的抑制作用。当Huh7细胞经miR-21 mimics处理后,Wnt4的mRNA及蛋白质表达均显著上升,且miR-21过表达可逆转DKK-1对Wnt4和细胞增殖的抑制作用。另外,miR-21过表达还上调了PTEN的mRNA及蛋白表达,并可提升经bpv(100和200nmol/L)抑制的Huh7细胞增殖速率。结论 miR-21在Huh7细胞对顺铂的耐药中发挥作用,且通过活化Wnt信号通路及PTEN调节Huh7细胞增殖。     

兔慢性肝损害急性肝衰竭动物模型的建立

目的 建立新西兰兔的慢加急性肝衰竭(ACLF)动物模型.方法 将75只新西兰兔随机分为实验组(n=70)与对照组(n=5),实验组采用四氯化碳(CCl4)腹腔注射建立兔代偿性肝纤维化模型,每周2次,通过体质量及谷丙转氨酶(ALT)/谷草转氨酶(AST)改变调整药物剂量,共10周,获得48只肝纤维化新西兰兔,在此基础上将动物随机分为4组(n=12),Ⅰ组:继续腹腔注射CCl4,Ⅱ组:静脉注射D-氨基半乳糖(D-Gal)0.65 g/kg,Ⅲ组:静脉注射D-Gal 0.70 g/kg,Ⅳ组:静脉注射D-氨基半乳糖0.75 g/kg,观察各组动物的一般情况、生化指标及病理改变.结果 与对照组比较,肝纤维化实验组兔10周时ALT、AST、ALB、γ-谷氨酰转肽酶(GGT)、透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PC-Ⅲ)差异均有统计学意义(P＜0.05),可观察到肝纤维化的病理表现,出现典型的假小叶,在肝纤维化基础给予D-Gal后,Ⅰ组动物全部存活,生化指标轻度改变;Ⅱ组动物死亡率为33.3%(4/12),生化指标仅出现一过性的改变;Ⅲ组动物死亡率为83.3%(10/12),平均存活时间为(53.00±25.69)h,给药后12 h生化指标及临床表现出现改变,48 h达到高峰,病理显示肝脏大块坏死;Ⅳ组动物均死于肝衰竭,平均存活时间为(32.70±17.46)h,肝损害出现时间早,损伤剧烈.结论 对CCl4诱导的肝纤维化新西兰兔给予D-Gal急性攻击可建立ACLF模型.其中给予D-Gal 0.70 g/kg的模型稳定性好,能较大程度上的模拟临床上ACLF的病理生理过程.

Abstract：

Objective To establish an ideal animal model of acute-on-chronic liver failure (ACLF) in New Zealand white rabbits in order to provide a large animal model for further researches.Methods Totally 75 New Zealand rabbits were randomly divided into experimental group (n =70) and control group (n = 5 ). Rabbits in the experimental group were injected with CCl4 into the abdominal cavity twice every week and the doses of CCl4 were modified according to the index of liver function and the body weight, whereas those in the control group were treated with the same volume of saline. At the 10th week,48 New Zealand rabbits with hepatic fibrosis were randomly assigned to 4 groups and injected with CCl4 as before, D-Gal at a dose of 0. 65 g/kg body weight (BW), 0. 70 g/kg BW and 0. 75 g/kg BW, respectively. By observing and comparing the general state, survival time, biochemical indexes, and the histopathology, a method of establishing a stable animal model of acute hepatic failure was found. Results As compared with those in control group, the levels of ALT, AST, GGT, HA, LN and PC-Ⅲ in the experiment group were increased significantly, while the level of ALB was decreased at the end of 10 weeks. Typical features of hepatic fibrosis and the formation of pseudo-lobules were observed at the end of 10 weeks. After treatment with D-Gal, all rabbits in group Ⅰ survived with minimal changes in liver function tests. In group Ⅱ , there was a temporary hepatic injury, but no hepatic coma. Four of the 12 rabbits died (33. 3% ). In group Ⅲ , biochemical indexes changed obviously 12 h after the administration and hepatic injury reached its peak after 48 h. Ten of 12 rabbits were died of severe hepatic failure with a survival time of ( 53. 00 ± 25. 69) h. Histology of liver section revealed massive necrosis in nodules. In group Ⅳ , hepatic injury occurred early and severely. All the rabbits died of severe hepatic failure with a survival time of (32. 70 ± 17. 46) h. Conclusion The experimental model of ACLF could be established by injected with D-Gal in New Zealand rabbits with hepatic fibrosis, induced by CCl4 intraperitoneal injection for 10 weeks.The one induced by 0. 70 g/kg of D-galactosamine was more stable and showed similar clinical pathophysiological changes in human beings. So it can be a good experimental platform for studies of ACLF.     

肝移植治疗成人郎格罕细胞组织细胞增生症伴肝脏累及的护理

目的探讨肝移植治疗成人郎格罕细胞组织细胞增生症(langerhans cell histiocytosis,LCH)伴肝脏累及的观察护理。方法回顾性分析1例成人LCH伴肝脏累及患者在我院行肝移植术期间的护理特点。结果患者术后肝功能恢复良好,术前存在的尿崩症得到有效控制,术后初期出现的腹泻、发热、电解质紊乱等在治疗护理后恢复正常,术后42天患者出院,定期随访。结论肝移植治疗LCH伴肝脏累及患者比较罕见,除了做好普通肝脏移植术后常规护理外,还应加强对疼痛、肢体活动度、尿量及水电解质等方面的观察护理,并给予饮食、运动、服药等健康指导。     

兔慢性肝损害急性肝衰竭动物模型的建立

Objective To establish an ideal animal model of acute-on-chronic liver failure (ACLF) in New Zealand white rabbits in order to provide a large animal model for further researches.Methods Totally 75 New Zealand rabbits were randomly divided into experimental group (n =70) and control group (n = 5 ). Rabbits in the experimental group were injected with CCl4 into the abdominal cavity twice every week and the doses of CCl4 were modified according to the index of liver function and the body weight, whereas those in the control group were treated with the same volume of saline. At the 10th week,48 New Zealand rabbits with hepatic fibrosis were randomly assigned to 4 groups and injected with CCl4 as before, D-Gal at a dose of 0. 65 g/kg body weight (BW), 0. 70 g/kg BW and 0. 75 g/kg BW, respectively. By observing and comparing the general state, survival time, biochemical indexes, and the histopathology, a method of establishing a stable animal model of acute hepatic failure was found. Results As compared with those in control group, the levels of ALT, AST, GGT, HA, LN and PC-Ⅲ in the experiment group were increased significantly, while the level of ALB was decreased at the end of 10 weeks. Typical features of hepatic fibrosis and the formation of pseudo-lobules were observed at the end of 10 weeks. After treatment with D-Gal, all rabbits in group Ⅰ survived with minimal changes in liver function tests. In group Ⅱ , there was a temporary hepatic injury, but no hepatic coma. Four of the 12 rabbits died (33. 3% ). In group Ⅲ , biochemical indexes changed obviously 12 h after the administration and hepatic injury reached its peak after 48 h. Ten of 12 rabbits were died of severe hepatic failure with a survival time of ( 53. 00 ± 25. 69) h. Histology of liver section revealed massive necrosis in nodules. In group Ⅳ , hepatic injury occurred early and severely. All the rabbits died of severe hepatic failure with a survival time of (32. 70 ± 17. 46) h. Conclusion The experimental model of ACLF could be established by injected with D-Gal in New Zealand rabbits with hepatic fibrosis, induced by CCl4 intraperitoneal injection for 10 weeks.The one induced by 0. 70 g/kg of D-galactosamine was more stable and showed similar clinical pathophysiological changes in human beings. So it can be a good experimental platform for studies of ACLF.     

杂合子亲属活体肝移植治疗儿童BCKDHB基因新复合突变枫糖尿病1例并文献复习

目的 总结枫糖尿病的杂合子亲属活体肝移植治疗效果.方法 1例男性3岁患儿,因发现枫糖尿病半年于2017年7月5日入院.患儿空腹状态出现阵发性口齿不清、下肢运动功能障碍半年,枫糖样体味明显,语言发育稍迟缓,无其他生长发育异常和智力缺陷.血清支链氨基酸(BCAA)检查示亮氨酸684μmol/L,缬氨酸559μmol/L,经...