人细胞系生物人工肝支持系统治疗慢加急性肝衰竭患者的初步研究

目的利用自行构建的混合生物人工肝支持系统,探讨其治疗肝衰竭患者的安全性和有效性。方法采用转染人肝再生增强因子（hALR）的HepG2细胞为生物材料构建生物反应器,利用慢加急性肝衰竭患者作为治疗对象,随机分组进行混合生物人工肝治疗及普通血浆置换治疗。结果治疗组6例患者中,4例经住院治疗1个月后临床好转出院,1例在治疗结束20d后因肝性脑病死亡,1例出院后1个月因肝肾综合征死亡,1例在恢复后1年因消化道出血死亡。对照组6例患者中存活2例,1例肝移植,3例因肝衰竭死亡。结论自行构建的混合生物人工肝支持系统功能良好,初步论证有一定的安全性和有效性。     

人肝细胞离线混合生物人工肝支持系统的构建与实验研究

目的 构建人肝细胞离线混合生物人工肝支持系统,探讨其治疗肝衰竭患者离体血浆的可行性,为开展离线生物人工肝治疗肝衰竭患者奠定基础.方法 采用转染人肝再生增强因子（hALR）基因的Hep G2细胞为生物材料,将细胞培养于费森尤斯血浆滤过器（Psu-2S）中,构建生物反应器.利用慢性乙型病毒性肝炎基础上发生的慢加急性肝衰竭患者血浆置换后离体血浆作为治疗对象,先行2h血浆胆红素吸附治疗,然后应用构建的生物人工肝系统对其进行实验性治疗.结果 生物反应器建立过程顺利,无菌技术达到要求,细胞培养液离心检测未见细胞漏出,生物反应器外腔液中细胞活力平均达95.6%,细胞增殖旺盛.肝衰竭患者血浆在胆红素吸附治疗前后分别为（176.19±54.14）μmol/L,（50.10±16.85）μmol/L,差异有统计学意义（t=8.32,P=0.0002）,白蛋白、尿素氮、血糖等指标在胆红素吸附前后差异无统计学意义.血浆在生物人工肝治疗前后,胆红素分别为（50.10±16.85）μmoL/L,（30.27±15.02）μmol/L,差异有统计学意义（t=13.19,P=0.000）;同时尿素氮（UERA）明显升高（t=15.4,P=0.000）;血糖（GLU）明显降低（t=5.67,P=0.0013）;白蛋白治疗前后分别为（6.13±2.04）g/L及（7.19±2.42）g/L,差异无统计学意义（t=1.73,P=0.134）.结论 自行构建的混合生物人工肝支持系统细胞繁殖活性良好,对慢加急性肝衰竭患者血浆胆红素有较好的吸附及代谢作用并具有一定的蛋白合成功能.     

人工肝支持系统治疗重型病毒性肝炎的研究

目的 研究人工肝支持系统治疗重型病毒性肝炎的方法及疗效。方法 利用Baxter-550人工肾和Biologic-DT system进行人工肝支持系统（ALSS）治疗肝炎患者83例，其中重型肝炎患者66例。结果 治疗后平均胆红素、ALT、AST、BUN、Cr及内毒素较治疗前均有明显下降。66例重型肝炎患者中治愈好转31例（47．0％），死亡或自动出院35例（53．0％），对照组181例重型肝炎患者中治愈好转50例（27．6％），死亡或自动出院131例（72．4％）。结论 应用ALSS治疗重型肝炎患者具有一定疗效。     

非生物人工肝支持系统的临床应用现状及进展

肝衰竭作为临床常见的严重肝病症候群,病死率极高,其治疗包括内科综合治疗、非生物型人工肝支持系统治疗、肝移植和生物干细胞移植等。作为早中期肝衰竭患者过渡到肝移植的重要治疗方法,非生物型人工肝目前已得到国内外的认可与广泛应用。本文重点对常用的非生物型人工肝治疗方式进行综述。     

人肝细胞离线混合生物人工肝治疗慢加急性肝衰竭一例

应用转染人肝再生增强因子（hALR）基因的HepG2细胞筛选出一株肝细胞作为生物材料,并建立生物人工肝支持系统,经检测具有良好生物合成及转化功能。我们采用离线混合生物人工肝治疗慢加急性肝衰竭1例,报道如下。     

猪逆转录病毒与生物人工肝的使用

生物型人工肝(bioartificial liver,BAL)或组合型生物人工肝(hybridbioartificial liver,HBL),是目前国内外公认最接近自然肝脏,功能也最全面的人工肝脏支持系统[1].所谓生物型人工肝,就是在整个治疗系统中包含有活的肝细胞或组织,因此具有相应生物活性的人工肝支持系统.在生物人工肝领域,因为猪解剖、生理特点与人类相似,猪细胞来源广泛,经济适用,因此目前应用最多的生物部分是猪肝脏细胞.但目前已知,作为异种细胞猪肝细胞内存在着猪内源性逆转录病毒(porcine endogenousretrovirus,PERV),有引起生物人工肝治疗患者PERV感染的危险.因此,本文就猪内源性逆转录病毒及猪源性生物人工肝的安全性问题作一简要总结.     

生物人工肝支持系统中溶解氧控制问题的研究进展

生物人工肝支持系统借助体外的机械、生化装置,清除患者体内积蓄的各种有害物质,补充有利于肝细胞再生的必需物质,改善内环境,暂时替代患者病肝的部分功能,为肝衰竭患者过渡到自身肝细胞再生或肝移植赢得宝贵的时间.在生物人工肝支持系统(BALSS)中,生物反应器是整套系统的核心.对生物反应器中多参数如溶解氧(DO)、pH和温度的有效控制是保障肝细胞生长和功能发挥的基础,其中溶解氧对细胞生长尤为重要.文中对反应器中增加溶解氧浓度的方法、溶解氧的检测和溶解氧的控制算法进行了综述,并介绍了适用于生物人工肝溶解氧控制的自适应预测算法.     

同一机构5年原位肝移植726例术后并发肝动脉血栓形成14例资料回顾

背景：虽然肝移植技术已经成熟,但肝动脉血栓形成仍是造成肝移植后移植物丢失的重要原因之一,肝动脉血栓形成如果不能及早发现处理,只有再次肝移植才能挽救患者生命。 目的：总结原位肝移植后并发肝动脉血栓形成的治疗体会。 方法：中山大学附属第一医院器官移植中心从2004-01/2009-12共实施726例成人尸肝移植,共14例患者经造影证实在肝移植后出现肝动脉血栓形成,回顾性分析以上14例患者的临床资料。 结果与结论：肝动脉血栓形成的发生率为1.9%(14/726),发生的平均时间为移植后10 d(1~41 d)。14例肝动脉血栓形成患者中,6例表现为急性的肝功能恶化,4例表现为胆漏,1例表现为肝脓肿,3例无明显临床症状。3例行急诊肝动脉再血管化,2例行肝动脉溶栓治疗,3例行肝动脉再血管化联合肝动脉局溶栓治疗,6例行再次肝移植。本组肝动脉血栓形成相关的死亡率为42.9%(6/14),其中2例行肝动脉再血管化后因胆道坏死、肝功能衰竭死亡;1例溶栓后再次血栓形成并发多器官功能衰竭死亡;1例肝动脉再血管化联合肝动脉溶栓后因肾功能衰竭、严重感染死亡;2例再次移植后早期因严重感染死亡。8例患者康复出院,并常规随访18~66个月,其中2例患者分别于肝移植后18,29个月因肝癌复发死亡,以上患者随访过程中移植肝功能正常,肝动脉畅通。提示肝动脉血栓形成是肝移植后的严重并发症,在造成不可逆的胆道和肝实质损伤前,尽早恢复肝动脉血流可以避免再次肝移植。     

121例人工肝治疗肝衰竭病例流行病学分析

目的:分析采取人工肝治疗的肝衰竭患者流行病学特点.方法:收集本院2019-2021年采取人工肝治疗的肝衰竭病例.对其一般资料、入院方式、入住科室、住院时间、临床分型、肝衰竭病因、人工肝治疗次数等进行统计分析.结果:121例人工肝治疗的肝衰竭患者平均年龄49.86±13.17岁,40-50岁年龄组占比最高(24.79％),男女之比3.17:1;肝衰竭的临床分型以慢加急性肝衰竭为主(71.07％),其次为慢性肝衰竭(12.40％)、急性肝衰竭(11.57％)和亚急性肝衰竭(4.96％),其中乙型病毒性肝炎为慢加急性肝衰竭的主要病因;平均住院时间为23.26±11.71 d,其中慢性肝衰竭患者平均住院时间最长为26.53±12.73 d;平均人工肝治疗次数4.21±3.00次,血浆置换治疗模式使用最多486次,其次为血浆灌流、血液滤过、双重血浆分子吸附和血浆透析滤过.结论:进行人工肝治疗的肝衰竭患者在性别和年龄上存在差异,临床分型上以慢加急性肝衰竭为主,乙型病毒性肝炎仍为主要病因.     

猪逆转录病毒与生物人工肝的使用

生物型人工肝(bioartificial liver，BAL)或组合型生物人工肝(hybfidbioartifieial liver，HBL)，是目前国内外公认最接近自然肝脏，功能也最全面的人工肝脏支持系统。所谓生物型人工肝，就是在整个治疗系统中包含有活的肝细胞或组织，因此具有相应生物活性的人工肝支持系统。在生物人工肝领域，因为猪解剖、生理特点与人类相似，猪细胞来源广泛，经济适用，因此目前应用最多的生物部分是猪肝脏细胞。但目前已知，作为异种细胞猪肝细胞内存在着猪内源性逆转录病毒(porcine endogenousretrovirus，PERV)，有引起生物人工肝治疗患者PERV感染的危险。因此，本文就猪内源性逆转录病毒及猪源性生物人工肝的安全性问题作一简要总结。     

中国人肝细胞系1运用于生物人工肝的生物代谢功能

背景：前期研究发现,中国人肝细胞系1细胞分化程度高且生物代谢功能良好, 并且中国人肝细胞系1细胞组织学上来源于正常肝组织,较其他来源于肿瘤源性的肝细胞系更为安全。 目的：探讨中国人肝细胞系中国人肝细胞系1细胞在混合型生物人工肝中的生物代谢功能。 方法：15只食蟹猴随机分成对照组(n=5)和治疗组(n=10),均建立急性肝功能衰竭模型,治疗组接受以全接触灌流型生物反应器接种微载体微重力中国人肝细胞系1细胞建立的人源细胞混合型生物人工肝进行治疗。 结果与结论：急性肝功能衰竭食蟹猴血清谷氨酸转氨酶、总胆红素、总胆汁酸、尿素氮、肌酐、血氨均明显上升,而白蛋白、Fischer指数则显著下降;人源细胞混合型生物人工肝治疗后,急性肝功能衰竭食蟹猴血清谷氨酸转氨酶、总胆红素、总胆汁酸、尿素氮、肌酐、血氨和白蛋白均恢复。提示中国人肝细胞系1细胞在混合型生物人工肝中生物代谢功能良好,表现出良好的肝特异性生物合成及生物代谢功能。     

胸腺法新在肿瘤患者肝移植术后应用的安全性研究

目的探讨胸腺法新在肝脏移植患者中应用对急性排异反应发生率的影响及相关不良反应,评估其应用的安全性。方法选取2008年1月后超＂杭州标准＂的13例肝脏恶性肿瘤患者作为治疗组,肝移植术后1个月开始应用胸腺法新。应用方案为1.6 mg皮下注射,1次/d,应用2周后改为2次/周,连续应用2年。随机选取2008年1月前肝移植患者13例作为对照组,对照组未应用胸腺法新,免疫抑制方案等与治疗组相同,观察比较应用治过程中相关副作用及急性排异反应的发生情况。结果随访期间两组间肝功能恢复情况及排斥反应发生率均无明显统计学差异,胸腺法新治疗组和对照组各1例患者发生急性排异反应,治疗组急性排异反应患者经停用胸腺法新并调整免疫抑制剂后肝功能恢复正常;对照组即行排异反应患者经激素冲击治疗后得以纠正。两组比较急性排异反应发生率无显著差异（P〉0.05）,治疗组未观察到其他该药物相关不良反应。结论肝脏移植患者应用胸腺法新不增加急性排异反应的发生率,在肝脏移植患者中应用是安全的。     

生物型人工肝支持系统研究进展

暂时的功能替换和加强支持在抢救终末期器官衰竭病人中有重要作用。透析技术已常规用于急慢性肾功能衰竭治疗。但肝脏功能极为复杂 ,人工模拟或部分替换其功能一直都只是临床医生和生物工程学家的梦。随着细胞生物学和材料科学研究的飞速发展 ,结合生物医用材料和肝细胞体外培养技术而设计的体外灌流装置能替代人类肝脏部分功能 ,此即所谓“组合型”或生物型人工肝支持系统。最近 ,许多类型的人工肝支持系统已进入临床前试验。本文就生物型人工肝的基础和临床应用展开综述     

Artificial liver support: future aspects

Liver transplantation and blood purification therapy, including plasmapheresis, hemodiafiltration, and bioartificial liver support, are available to treat patients with severe liver failure. The two mainstream systems developed for bioartificial liver support are extracorporeal whole-liver perfusion (ECLP) and the bioreactor system (BIS). We developed a method of plasma cross-perfusion, in which plasma is exchanged between the blood circuit of the patient and that of a hepatic function unit, that is, whole liver or a bioreactor through which immunologically free whole human blood is perfused. From the aspects of efficacy and safety, the best system of bioartificial liver support for clinical use is considered to be ECLP in cross plasma perfusion. However, a social objection about zoonosis has consistently been raised, with controversy surrounding the use of xenogeneic organs for human treatment, and this might be a final obstacle to the development of system efficacy. The combination therapy of hemodiafiltration with the administration of human serum albumin and anticoagulant factors can minimize the economic and medical resource costs through the development of transgenic livestock that secrete human pharmaceuticals systemically. It is possible that this therapy will become the most practical treatment for patients with severe hepatic failure.     

Study of severe hepatitis treated with a hybrid artificial liver support system

Artificial liver support system (ALSS) has been used to treat hepatic failure and has significantly decreased the mortality. TECA hybrid artificial liver support system (TECA-HALSS), which combines the hollow fiber bioreactor with a plasma exchange circuit, was used to assess the efficacy, safety and feasibility in treating severe hepatitis patients. The hybrid artificial liver support system (HALSS) consists of a bioreactor containing more than 5 x10(9) porcine hepatocytes and plasma exchange device. Fifteen patients with severe hepatitis were treated with this hybrid system. All patients experienced a reduction in symptoms such as fatigue, abdominal distention or ascites. After each treatment serum total bilirubin decreased markedly while prothrombin activity increased. There were ten patients whose progress of hepatocyte necrosis was stopped after HALSS treatment, and finally they recovered completely. One patient received liver transplantation after HALSS therapy and survived. No serious adverse events were noted in the fifteen patients.     

Early experiences with a porcine hepatocyte-based bioartificial liver in acute hepatic failure patients

Orthotopic liver transplantation (OLT) is the only effective therapeutic modality in severe acute hepatic failure (AHF). The scarcity of organs for transplantation leads to an urgent necessity for temporary liver support treatments in AHF patients. A hepatocyte-based bioartificial liver (BAL) is under investigation with the main purpose to serve as bridging treatment until a liver becomes available for OLT, or to promote spontaneous liver regeneration. We developed a novel radial-flow bioreactor (RFB) for three-dimensional, high-density hepatocyte culture and an integrated pumping apparatus in which, after plasmapheresis, the patient's plasma is recirculated through the hepatocyte-filled RFB. Two hundred thirty grams of freshly isolated porcine hepatocytes were loaded into the RFB for clinical liver support treatment. The BAL system was used 8 times in supporting 7 AHF patients in grade III-IV coma, all waiting for an urgent OLT Three patients with no history of previous liver diseases were affected by fulminant hepatic failure (FHF) due to hepatitis B virus, 3 by primary non-function (PNF) of the transplanted liver, and one by AHF due to previous abdominal trauma and liver surgery. Six out of 7 patients underwent OLT following BAL treatment(s), which lasted 6-24 hours. All patients tolerated the procedures well, as shown by an improvement in the level of encephalopathy, a decrease in serum ammonia, transaminases and an amelioration of the prothrombin time, with full neurological recovery after OLT Our initial clinical experience confirms the safety of this BAL configuration and suggests its clinical efficacy as a temporary liver support system in AHF patients.     

Function of a new internal bioartificial liver: an in vitro study

The goal of this study was to determine whether a new internal bioartificial liver utilizing porcine hepatocytes can perform detoxification and other metabolic functions. Such a system might aid in treating patients with moderate to severe liver failure and prolong patient survival until a matching organ is found for transplantation. Porcine hepatocytes were attached to a microcarrier and an internal artificial liver was constructed by perfusing the hepatocytes into a polysulfon hollow fiber. The 4 experimental groups were: (a) control group, (b) microcarrier group, (c) hollow fiber group, and (d) internal bioartificial liver group. Viability of hepatocytes, alanine aminotransferase (ALT) and lactate dehydrogenase (LD) activities in the medium, urea production, diazepam transformation, protein synthesis, and glucose-6-phosphatase activity of cells were monitored during a 7-day culture period. Viability of porcine hepatocytes in the internal bioartificial liver group was maintained at >80% during the culture period, and alanine aminotransferase and lactate dehydrogenase activities did not fluctuate significantly. These enzyme activities were significantly lower in the internal bioartificial liver group than in the control or microcarrier groups. Urea production, diazepam transformation, [3H]-leucine incorporation, and glucose-6-phosphatase activity were significantly higher in the internal bioartificial liver group than in the control and hollow fiber groups. These results show that the new internal bioartificial liver produces small amounts of ALT and LD and exhibits detoxification and protein synthetic functions.     

Assay to detect inhibitory substances in serum of patients with acute liver failure

Patients with acute liver failure accumulate toxic substances in the circulation which may impair recovery of hepatic function. The aim of this study was to test an in vitro assay to detect inhibitory substances in the serum of patients with acute liver failure. Human liver-derived HepG2 cells were incubated for 24h in 96 well plates (30,000 cells/well) with sera (10%) from 24 patients with acute liver failure due to paracetamol overdose or NANB hepatitis and 11 normal controls. DNA synthesis was determined from the incorporation of 3H-thymidine and cell viability by the metabolism of the tetrazolium dye MTS. HepG2 cells exposed to acute liver failure sera incorporated significantly less 3H-thymidine (median 30% of control, range 0.2-169%) than normal sera (100%, 76-133%, p=0.002). Cell viability was also reduced (75%, 33-112% vs 100%, 96-105%, p<0.001). There was no correlation between these values and patient outcome or levels of plasma TNF-alpha or serum interferon-gamma. The assay detected inhibitory substances in sera of patients with acute liver failure and could be used to monitor the use of liver support systems.