胎肝前体细胞脾移植对大鼠急性肝功能衰竭的实验研究

目的 :探讨经脾内同种异体移植培养的原代胎肝前体细胞与成体肝细胞悬液对大鼠药物性肝衰竭的疗效 ,并观察脾内移植肝细胞的生物学特性。方法 :采用 D-氨基半乳糖 ( D- gal)建立大鼠急性肝衰竭模型 ,2 4 h后随机分为三组进行治疗。 A组 :经脾内移植体外培养 7d的肝细胞 2× 1 0 7;B组 :经脾内移植肝细胞悬液 2× 1 0 7;C组 :经脾内注射生理盐水1 ml。观察受体大鼠的存活率、肝脏功能和病理变化及移植肝细胞的生物学特性。结果 :A组、B组大鼠存活率 ( 77%、5 9% )与 C组大鼠存活率 ( 2 2 % )相比具有显著性差异 ,肝功能各项指标均有明显改善 ,A、B组与 C组大鼠的肝功能改变方面有统计学差异。经 HE和 PAS染色证实 ,移植的肝细胞在受体脾内结构和功能保持较好。结论 :经脾内移植的培养的原代胎肝前体细胞和肝细胞悬液均能提高大鼠药物性肝衰竭的存活率、改善肝功能及肝脏组织病理变化 ,但培养原代胎肝前体细胞优于成体肝细胞悬液。     

胃选择性迷走神经切断对大鼠肝再生的影响——肝细胞体积、数量和DNA含量的研究

探讨胃选择性迷走神经切断对大鼠再生肝细胞体积、数量和ＤＮＡ含量的影响。分别测定大鼠胃选择性迷走神经切断加肝叶大部分切除及肝叶大部分切除术后７ ｄ 再生肝ＤＮＡ含量、肝细胞数和体积。用多功能显微图像测量仪测定切片中肝细胞和肝细胞核面积及肝细胞核直径。按体视学方法测算整个肝脏的肝细胞数。用细胞分光光度技术测定切片中单个肝细胞ＤＮＡ 含量。结果表明, 胃选择性迷走神经切断加肝叶大部分切除组大鼠肝细胞ＤＮＡ含量、数量及体积均较单纯肝叶切除组增加 （Ｐ＜ ０．０５）。提示胃选择性迷走神经切断对大鼠肝再生过程有一定的促进作用。     

胃选择性迷走神经切断对大鼠肝再生的影响

探讨胃选择性迷走神经切断对大鼠再生肝细胞体积、数量和DNA含量的影响。分别测定大鼠胃选择性迷走神经切断加肝叶大部分切除及肝叶大部分切除术后7 d再生肝DNA含量、肝细胞数和体积。用多功能显微图像测量仪测定切片中肝细胞和肝细胞核面积及肝细胞核直径。按体视学方法测算整个肝脏的肝细胞数。用细胞分光光度技术测定切片中单个肝细胞DNA含量。结果表明,胃选择性迷走神经切断加肝叶大部分切除组大鼠肝细胞DNA含量、数量及体积均较单纯肝叶切除组增加(P＜0.05)。提示胃选择性迷走神经切断对大鼠肝再生过程有一定的促进作用。     

大鼠原位部分肝移植模型的建立及技术改进

目的建立简便实用的大鼠原位部分肝移植模型.方法用120只健康SD大鼠,体重为(285±55)g,随机分为供体组和受体组,每组60只.取供体大鼠肝脏左内叶、中叶、尾叶和右叶为移植物,占受体大鼠肝重的59.1%～64.3%.门静脉、肝下下腔静脉采用改良的二袖套法吻合.观察受体大鼠术后存活率、死亡原因、移植物光镜和电镜下表现以及肝组织再生情况.结果24h存活率93.33%,7d存活率68.33%,光镜下发现肝细胞增生活跃,电镜下发现肝细胞超微结构变化不明显,7d后肝移植重量达原受体大鼠肝重的88.53%.结论改进的大鼠原位部分肝移植术操作简便,手术成功率高,重复性好,可用于原位部分肝移植的实验研究.     

混合细胞共微囊化对肝细胞功能的支持作用

目的观察大鼠肝细胞、转基因肝星状细胞株HGF／CFSC和／或大鼠骨髓来源Thy-1^＋β2M^-细胞（BDTC）共微囊化对肝细胞生物学活性的支持,及腹腔移植混合细胞微囊对急性肝衰竭大鼠肝功能的改善作用。方法利用微囊发生器制备含肝细胞或混合细胞的微囊,依微囊内包裹细胞种类不同,分为微囊化肝细胞组、微囊化肝细胞＋CFSC／HGF组）和微囊化肝细胞＋CFSC／HGF＋BDTC组,通过观察囊内细胞形态和体外培养测定培养液中白蛋白和尿素的分泌,判断各组囊内肝细胞活性和功能的维持;将90％肝大部切除所致的急性肝衰竭大鼠按照移植微囊种类不同分为空囊对照组和上述3个实验组（每组10只）,观察腹腔植入后不同时间大鼠的一般状况、存活时间、血生化改变、肝组织再生及微囊化移植物的组织学特征。结果与单独肝细胞微囊者相比,混合细胞微囊内肝细胞存活时间超过1倍,培养液中白蛋白分泌和尿素合成量明显增加（均P〈0．01）;与对照组相比,微囊化肝细胞或微囊化混合肝细胞移植后,急性肝衰竭大鼠的肝功能显著改善、存活率明显提高（10／10 vs 1／10）,其肝组织再生完全;移植21—42d时,部分微囊附着于肝脏表面并出现血管化,微囊表面存在不同程度的纤维化,微囊内仍有存活的细胞,以微囊化混合肝细胞组优于微囊化肝细胞组。结论混合细胞共微囊化能明显改善囊内肝细胞的存活寿命、形态和功能的维持,微囊化混合肝细胞腹腔移植对促进急性肝衰竭大鼠的肝功能恢复具有显著作用。     

肝细胞脾内移植治疗大鼠肝衰竭

目的:优化急性肝衰竭模型的构建方法及探讨经脾内同种异体肝细胞移植治疗大鼠急性药物性肝衰竭的疗效,并观察脾内移植肝细胞的生物学活性。方法:采用D-氨基半乳糖(D-gal)建立大鼠急性药物性肝衰竭模型。腹腔注射不同剂量的(1.0,1.2,1.4,1.6g/kg)D-gal后,以肝功能指标、病理形态学、死亡率等综合评估肝脏细胞损伤的状态,确定模型构建的最佳方案。造模后24h,随机分为2组进行治疗,Ⅰ组:经脾内移植肝细胞悬液1ml(4.4×107个);Ⅱ组:经脾内移植不含肝细胞的培养液1ml。观察大鼠1周的存活率、肝脏功能和病理变化及移植肝细胞的生物学活性。结果:①不同剂量的D-gal可导致不同程度的肝损伤,而以1.2g/kgD-gal造模组的大鼠死亡率适中,肝功能损害及其肝脏病理组织学改变均符合临界急性肝衰竭的程度,较好地模拟了急性肝衰竭的病理生理改变且稳定性较好。②移植后3d,Ⅰ组与Ⅱ组大鼠存活率相比具有显著性差异(P<0.01);肝功能指标:丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)和总胆红素(TBiL)均有明显改善(P<0.01);肝组织病理切片显示:Ⅰ组的肝组织损伤程度明显比Ⅱ组轻。经HE和PAS染色证实,移植的肝细胞在受体脾内结构和功能保持较好。结论:①D-gal1.2g/kg腹腔注射可较好构建SD大鼠急性肝衰竭模型。②经脾内肝细胞移植能明显提高药物性肝衰竭大鼠的存活率、改善肝功能及肝组织病理变化。     

人胎肝细胞对大鼠暴发性肝功能衰竭影响的初步研究

本文报道用D-氨基半乳糖所致大鼠暴发性肝衰竭作模型,观察人胎肝细胞对其影响。结果显示,腹腔内注射人胎肝细胞悬液和胞质液均能显著提高肝衰竭大鼠的存活率,两者间比较无显著差别。人胎肝细胞质液可减轻中毒大鼠内毒素血症,降低肝内丙二醛水平,阻止肝坏死,促进肝再生。实验结果提示人胎肝细胞治疗肝衰竭可能主要通过胞质液中的生物活性物质起作用。     

大鼠冷保存供肝肝移植模型的建立及评价

目的 建立稳定的大鼠供肝冷保存肝移植模型,为冷保存再灌注损伤及再生研究奠定基础.方法 大鼠原位肝移植模型参照Kamada的双袖套法并加以改进.雄性SD大鼠分为4组,观察不同的冷保存时间对大鼠生存率及损伤与再生状况的影响.结果 模型的平均无肝期为15 min左右,肝移植对照组、冷冻保存8、12、16 h组术后168 h存活率分别为100%、80%、50%、20%.冷冻保存12 h组与肝移植对照组相比,大鼠存活率明显降低(P＜0.05).冷冻保存12 h组肝脏组织术后24 h可观察到明显的片状坏死,继而肝细胞有丝分裂相显著增多.结论 对SD大鼠而言,冷保存12 h的供肝肝移植是研究肝脏损伤与再生修复的良好模型.     

大鼠肝叶切除后肝细胞再生相关蛋白的蛋白质组学分析

目的 通过对肝叶切除后6、24 h的肝细胞与正常肝细胞的蛋白质组分析,筛选出与肝细胞再生相关的蛋白质.方法 选择雄性SD大鼠9只(体质量200 g),分3组,6 h组、24 h组和对照组,每组3只,建立大鼠70%肝叶切除模型.6 h组在肝叶切除后6 h分离肝细胞,24 h组在肝叶切除后24 h分离肝细胞,对照组为正常肝细胞.裂解液裂解细胞提取总蛋白,运用蛋白组学相关技术分析3组细胞的蛋白组学差异.结果 总共得到47个差异蛋白质点,鉴定出42个.结论 我们的数据显示:24 h组与对照组和6 h组都有重叠,而6 h组和对照组重叠部分较少,推测其可能的原因为6 h组肝细胞处于肝再生的启动阶段,与对照组差异较大,而24 h组,肝再生已启动,各种维持肝细胞正常功能的蛋白质也开始表达.     

大鼠肝脏缺血再灌注合并肝部分切除诱导肝再生模型中EGFR及CyclinD1的表达

目的 观察大鼠肝脏缺血再灌注合并肝再生诱导模型中EGFR及CyclinD1的表达情况。方法 采用预先保留门静脉转流肝叶（约占全肝70％）,阻断部分肝叶（约占全肝30％）血供,到预定观察终点后重新开放血供灌注缺血肝叶,同时切除门静脉转流肝叶（70％肝切除）模型,通过RT－PCR和免疫组化法分析EGFR及CyclinD1大鼠肝脏缺血再灌注对肝细胞再生功能影响过程中的变化。结果 大鼠术后12、24hCyclinD1mRNA及此后相应的蛋白表达水平随着缺血时间的延长而显著降低。术后12h内EGFR蛋白表达水平随着缺血时间的延长而显著降低。结论 术后EGFR以及CyclinD1表达降低可能是肝脏缺血再灌注影响大鼠肝再生的原因之一。     

探讨脂肪肝供肝在肝脏移植中的价值

目的探讨应用不同脂肪沉着程度供肝对肝移植患者预后的影响。方法选取64例进行肝移植术患者,依据供肝浸润程度,随机分为轻度脂肪肝组、中度脂肪肝组、重度脂肪肝组、无脂肪肝组共四组,每组各16例,比较四组患者肝功能(主要观察ALT、AST指标)、入住ICU的平均时间、1年移植物成活率、移植物原发无功发生率。结果轻度脂肪肝组入住ICU的平均时间、ALT和AST水平、移植物原发无功发生率与无脂肪肝组接近;中度脂肪肝组入住ICU平均时间、ALT和AST水平均较轻度组和无脂肪肝组明显增高,P0.05;重度脂肪肝组入住ICU平均时间、ALT和AST水平、移植物原发无功发生率明显较前三组为高,P0.05。结论轻、中度肝脏脂肪浸入可作为肝移植供肝,重度脂肪肝不能作为供肝的选择目标。     

Microenvironment of Liver Regeneration in Liver Cancer

The occurrence and development of liver cancer are essentially the most serious outcomes of uncontrolled liver regeneration. The progression of liver cancer is inevitably related to the abnormal microenvironment of liver regeneration. The deterioration observed in the microenvironment of liver regeneration is a necessary condition for the occurrence, development and metastasis of cancer. Therefore, the use of a technique to prevent and treat liver cancer via changes in the microenvironment of liver regeneration is a novel strategy. This strategy would be an effective way to delay, prevent or even reverse cancer occurrence, development and metastasis through an improvement in the liver regeneration microenvironment along with the integrated regulation of multiple components, targets, levels, channels and time sequences. In addition, the treatment of "tonifying Shen (Kidney) to regulate liver regeneration and repair by affecting stem cells and their microenvironment" can regulate "the dynamic imbalance between the normal liver regeneration and the abnormal liver regeneration"; this would improve the microenvironment of liver regeneration, which is also a mechanism by which liver cancer may be prevented or treated.     

Liver

Alleviation of ischemia-reperfusion injury in rat liver donors by induction of exogenous hTERT gene;Effects of surgical technique on acute renal failure after orthotopic liver transplantation in patients with end-stage liver disease at high risk： a report of 90 eases;Timing for liver transplantation for chronic severe hepatitis;Analysis of bacterial variance and drug resistance after orthotopic liver transplantation;The influence of splenectomy on orthotopic liver transplantation and its management     

Liver

<正>209461 Intraoperative ligation of recipient’s portasystemic shunt in liver transplantation/Chen Litian(,Organ Transplant Center,Tianjin 1st Centr Hosp Tianjin 300192)…∥Chin J Gen Surg.-2009,25(4).-489～491Objective To investigate the clinical significance of ligating the portasystemic shunt confirmed by preoperative CT evaluation during orthotopic liver transplantation.Methods From January 2007 to August 2008,35 patients in Tianjin First Central Hospital underwent preoperative three-dimensional CT scan,among them 23 patients had spontaneous major portasystemic shunts,the other 12 patients did not have portasystemic shunts.16 out of the 23 cases with significant shunts underwent shunt ligation based on portal blood flow volume measured by intraoperative portal vein flowmetry.The shunt of the other 7 patients were left untreated.Results The portal blood flow in the 12 patients without portasystemic shunt judged by preoperative CT scanning were(1 101±70)ml/min.The shunts in 7 patients with portal blood flow greater than 1 000 ml/min were not ligated,that of the 16 patients with portal blood flow volume lower than 1 000 ml/min were ligated.The portal blood flow volume in those 16 patients before and after ligating the shunt were(657±112)ml/min and(1 136±161)ml/min,respectively(P<0.05).Postoperatively 2 patients suffered from portal vein thrombosis,among them 1 patients suffered from intermittent disturbance of consciousness,2 patients died within 3 months,with one died of respiratory failure from pulmonary aspergillus infection,one died of hepatic failure in 2 months after operation because of graft dysfunction.The other 19 patients with normal blood flow and well-functioning graft were alive.Conclusion The ligation of portasytemic shunt is mandatory in patients when pretransplant CT evaluation showing a major porto-systemic shunts and portal blood flow volume was less than 1 000 ml/min.5 refs,2 figs.     

Liver

<正>209293 Probing into indication of living-related Liver Transplantation for Wilson’s disease/Cheng Feng(Liver Transplant Center,1st Hosp Nanjing Med Univ,Key Lab Living Donor Liver Transplant,Minist Public Health,Nangjing 210029)…∥Chin J Surg.-2009,47(6).-437~440Objective To probe into indication of living-related liver transplantation(LRLT)for Wilson’s Disease.Methods From January 2001 to February 2007,thirty-seven living-related liver transplants were performed.A retrospective analysis was carried on outcome of those patients.The indications for LRLT were acute hepatic failure in 3 patients and chronic advanced liver disease in 32 patients including 13 patients with Wilsonian neurological manifestations.Two patients presented with severe Wilsoian neurological manifestations even though their liver functions were stable.According to the scoring system for evaluation of the neurological impairment in Wilsion disease based on neurological signs and functions(total score was 30),the pre-transplantation score of those patients with neurological manifestations was 15.9±4.3(n=15).Results Thirty-seven patients were followed up for 20-93 months.The survival rates of post-transplant patients and grafts at 1,3,and 5 year were 91.9%,83.8%,75.7%,and 86.5%,78.4%,75.7%,respectivly.Postoperative surgical complications occurred in 2 donors with bile leakage required drainage,in 2 recipients with hepatic thormbosis underwent retransplantation of cadeveric liver and in 1 recipient with hepatic stenosis required balloon dilatation.Neurological function was improved in all recipients and the score of posttransplantation at 6,12,18,24,and 30 months was 17.5±3.7(n=13);21.0±4.3(n=12);23.9±3.9(n=10);26.6±2.2(n=10)and 28.1±1.9(n=7)respectively.Conclusion Patients with acute hepatic failure or patients with severe liver disease unresponsive to chelation tratment should be treated with LRLT.Early transplantation in patients with an unsatisfactory response medical tratment may prevent irreversible neurologica     

脂肪肝患者血清肝纤维化指标分析

目的探讨脂肪肝患者血清肝纤维化指标特点，寻找可能引起脂肪肝肝纤维化改变的相关因素。方法记录脂肪肝组和正常对照组临床资料与血清生化指标以及肝纤维化指标，分析各项指标的异常及脂肪肝组肝纤维化指标及其他指标的相关关系。结果脂肪肝组和正常对照组血清生化指标和肝纤维化指标之间的比较：脂肪肝患者组血清FPG、TG、Tc、ALT、HA、LN、PCⅢ、CIV、CG与对照组比较差异有统计学意义（P〈0．05）。血清肝纤维化指标HA与年龄（≥50岁）、酗酒、男性、CG增高、ALT增高、高血压病有线性回归关系；血清肝纤维化指标LN与酗酒、肥胖、糖尿病有线性回归关系；血清肝纤维化指标CIV与高脂血症，ALT增高有线性回归关系；血清肝纤维化指标PcⅢ与年龄（I〉50岁）、高脂血症、高血压病有线性回归关系。结论通过多元线性回归分析显示：男性，年龄≥50岁，酗酒，肥胖，伴发高血压病、高脂血症、糖尿病，出现CG增高，ALT增高，可能是促进脂肪肝发生肝纤维化的危险因素。     

Liver

<正>209604 The suppressive effect of CD8+ CD28-regulatory T cells from spontaneous tolerance models on the acute rejection responses in rat liver transplantation/Chen Ning(陈宁,Dept Gastroenterol,Peking Univ Peop Hosp,Beijing 100044)…∥Chin J Organ Trans-plant. -2009,30(9). -524 ～526     

Liver

<正>209087 Clinical analysis of outcome of invasive fungal infection after kidney transplantation/Chen Guodong(陈国栋,Dept Transplant Surg,1st Hosp,Sun Yat sen Univ, Guangzhou 510080)…∥Chin J Organ Transplant. -2009, 30(10). -616～619 Objective To explore the outcome of invasive fungal infection in kidney transplantation and the influencing factors.     

Liver

Prevention and treatment of artery complication after liver transplantation by HBO , Clincal feature of cytomegabvirus pneumonia in orthtopic liver-transplantation and treatments, Risk factors of diabetes mellitus following orthotopic liver transplantation , Re-infection of hepatitis B virus after liver transplantation , Diagnosis and treatment of fungal infection after orthotopic liver transplantation , Two successful adult-to-adult living donor liver transplantation using dual grafts     

化肝汤1号治疗酒精性肝纤维化症状及肝功能改变

目的观察活血化瘀、软坚散结药配伍治疗肝纤维化时对患者主、次症状改变及肝功能的影响。方法将确诊为酒精性肝纤维化患者90例,随机分为三组,治疗组采用自制方剂化肝汤1号(醋鳖甲、黄芩、柴胡、赤芍等),对照组1采用凯西莱治疗,对照组2采用化肝汤1号和凯西莱联合治疗,疗程3个月。观察三组患者治疗前后主、次症状改善情况,血清天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、γ谷氨酰转肽酶(γ-GT)变化。结果三组治疗后的临床疗效比较差异无统计学意义(H=2.5897,P>0.05)。三组治疗后肝功能指标均有改善,与治疗前比较,差异有统计学意义(P<0.05)。结论化肝汤1号可有效地降低肝细胞的损害程度,促进受损细胞修复,从而改善肝功能。