人工肝治疗重型肝炎肝衰竭对近期预后的影响

目的:探讨人工肝治疗重型肝炎肝衰竭对近期预后的影响及其相关机制。方法:将120例重型肝衰竭患者根据治疗方法的不同分为治疗组与对照组各60例,两组均给予内科基础治疗,治疗组在此基础上加用人工肝治疗。结果:治疗组有效率90.0%,对照组有效率66.7%,两组比较差异有统计学意义。两组治疗前肠源性内毒素值、TBil及PTA值对比差异无统计学意义,治疗后Tbil值有明显下降,而肠源性内毒素与PTA值明显上升,组间对比差异有统计学意义。两组治疗前腹痛、腹胀、食欲不振评分对比差异无统计学意义,治疗后症状评分都明显下降,同时组间对比差异有统计学意义。结论:人工肝治疗重型肝炎肝衰竭能提高近期预后效果,改善肝功能与缓解临床症状,其机制的发挥可能与降低肠源性内毒素有关。

Artificial and bioartificial liver support:A review of perfusion treatment for hepatic failure patients

Liver transplantation and blood purification therapy,including plasmapheresis,hemodiafiltration,and bioartificial liver support,are the available treatments for patients with severe hepatic failure.Bioartificial liver support,in which living liver tissue is used to support hepatic function,has been anticipated as an effective treatment for hepatic failure.The two mainstream systems developed for bioartificial liver support are extracorporeal whole liver perfusion(ECLP)and bioreactor systems.Comparing various types of bioartificial liver in view of function,safety,and operability,we concluded that the best efficacy can be provided by the ECLP system.Moreover,in our subsequent experiments comparing ECLP and apheresis therapy,ECLP offers more ammonia metabolism than HD and HF.In addition,ECLP can compensate amino acid imbalance and can secret bile.A controversial point with ECLP is the procedure is labor intensive,resulting in high costs.However,ECLP has the potential to reduce elevated serum ammonia levels of hepatic coma patients in a short duration.When these problems are solved,bioartificial liver support,especially ECLP,can be adopted as an option in ordinary clinical therapy to treat patients with hepatic failure.

Treatment of hyperbilirubinemia with blood purification in China

The incidence of hyperbilirubinemia is high clinically, which is difficult to cure by medication, surgery or interventional therapies. Non-bioartificial liver is the main alternative in the blood purification for hyperbilirubinemia, which includes plasma exchange, hemoperfusion, hemodialysis, molecular adsorbent recycling system and so on. The research results and clinical experiences in China show that these methods are effective in lowering high levels of bilirubin with fewer side effects. The hyperbilirubinemias of different causes, with different complications or accompanying different diseases can be treated by different methods. Bioartificial liver, hybrid artificial liver support system and adsorbent membrane material have also been studied and their development in reducing hyperbilirubinemias has been achieved. This article gives a brief overview on the actuality and research improvement in blood purification for hyperbilirubinemia in China.

Preoperative risk factor analysis in orthotopic liver transplantation with pretransplant artificial liver support therapy

AIM： To assess the value of pre-transplant artificial liver support in reducing the pre-operative risk factors relating to early mortality after orthotopic liver transplantation （OLT）. METHODS： Fifty adult patients with various stages and various etiologies undergoing OLT procedures were treated with molecular adsorbent recycling system （MARS） as preoperative liver support therapy. The study included two parts, the first one is to evaluate the medical effectiveness of single MARS treatment with some clinical and laboratory parameters, which were supposed to be the therapeutical pre-transplant risk factors, the second part is to study the patients undergoing OLT using the regression analysis on preoperative risk factors relating to early mortality （30 d） after OLT. RESULTS： In the 50 patients, the statistically significant improvement in the biochemical parameters was observed （pre-treatment and post-treatment）. Eight patients avoided the scheduled Ltx due to significant relief of clinical condition or recovery of failing liver function, 8 patients died, 34 patients were successfully bridged to Ltx, the immediate outcome of this 34 patients within 30d observation was： 28 kept alive and 6 patients died. CONCLUSION： Pre-operative SOFA, level of creatinine, INR, TNF-α, IL-10 are the main preoperative risk factors that cause early death after operation, MARS treatment before transplantion can relieve these factors significantly.

Comparison between bioartificial and artificial liver for the treatment of acute liver failure in pigs

AIM： To characterize and evaluate the therapeutic efficacy of bioartificial liver （BAL） as compared to that of continuous hemodiafiltration （CHDF） with plasma exchange （PE）, which is the current standard therapy for fulminant hepatic failure （FHF） in Japan. METHODS： Pigs with hepatic devascularization were divided into three groups： （1） a non-treatment group （NT; n = 4）; （2） a BAL treatment group （BAL; n = 4）, （3） a PE ＋ CHDF treatment group using 1.5 L of normal porcine plasma with CHDF （PE ＋ CHDF, n -- 4）. Our BAL system consisted of a hollow fiber module with 0.2 i^m pores and 1 × 10^10 of microcarrier-attached hepatocytes inoculated into the extra-fiber space. Each treatment was initiated 4 h after hepatic devascularization. RESULTS： The pigs in the BAL and the PE ＋ CHDF groups survived longer than those in the NT group. The elimination capacity of blood ammonia by both BAL and PE ＋ CHDF was significantly higher than that in NT. Aromatic amino acids （AAA） were selectively eliminated by BAL, whereas both AAA and branched chain amino acids, which are beneficial for life, were eliminated by PE ＋ CHDF. Electrolytes maintenance and acid-base balance were better in the CPE ＋ CHDF group than that in the BAL group. CONCLUSION： Our results suggest that PE ＋ CHDF eliminate all factors regardless of benefits, whereas BAL selectively metabolizes toxic factors such as AAA. However since PE ＋ CHDF maintain electrolytes and acid-base balance, a combination therapy of BAL plus CPE ＋ CHDF might be more effective for FHF.

Artificial liver support in pigs with acetaminophen-induced acute liver failure

AIM To establish a reversible porcine model of acute liver failure(ALF) and treat it with an artificial liver system. METHODS Sixteen pigs weighing 30-35 kg were chosen and administered with acetaminophen(APAP) to induce ALF. ALF pigs were then randomly assigned to either an experimental group(n = 11), in which a treatment procedure was performed, or a control group(n = 5). Treatment was started 20 h after APAP administration and continued for 8 h. Clinical manifestations of all animals, including liver and kidney functions, serum biochemical parameters and survival times were analyzed. RESULTS Twenty hours after APAP administration, the levels of serum aspartate aminotransferase, total bilirubin, creatinine and ammonia were significantly increased, while albumin levels were decreased(P < 0.05). Prothrombin time was found to be extended with progression of ALF. After continuous treatment for 8 h(at 28 h), aspartate aminotransferase, total bilirubin, creatinine, and ammonia showed a decrease in comparison with the control group(P < 0.05). A cross-section of livers revealed signs of vacuolar degeneration, nuclear fragmentation and dissolution.Concerning survival, porcine models in the treatment group survived for longer times with artificial liver system treatment(P < 0.05). CONCLUSION This model is reproducible and allows for quantitative evaluation of new liver systems, such as a bioartificial liver. The artificial liver system(ZHj-3) is safe and effective for the APAP-induced porcine ALF model.

Evaluation of a novel hybrid bioartificial liver based on a multi-layer flat-plate bioreactor

AIM: To evaluate the efficacy and safety of a hybrid bioartificial liver (HBAL) system in the treatment of acute liver failure. METHODS: Canine models with acute liver failure were introduced with intravenous administration of D-galactosamine. The animals were divided into: the HBAL treatment group (n = 8), in which the canines received a 3-h treatment of HBAL; the bioartificial liver (BAL) treatment group (n = 8), in which the canines received a 3-h treatment of BAL; the non-bioartificial liver (NBAL) treatment group (n = 8), in which the canines received a 3-h treatment of NBAL; the control group (n = 8), in which the canines received no additional treatment. Biochemical parameters and survival time were determined. Levels of xenoantibodies, RNA of porcine endogenous retrovirus (PERV) and reverse transcriptase (RT) activity in the plasma were detected. RESULTS: Biochemical parameters were significantly decreased in all treatment groups. The TBIL level in the HBAL group was lower than that in other groups (2.19 ± 0.55 mmol/L vs 24.2 ± 6.45 mmol/L, 12.47 ± 3.62 mmol/L, 3.77 ± 1.83 mmol/L, P < 0.05). The prothrombin time (PT) in the BAL and HBAL groups was significantly shorter than the NBAL and control groups (18.47 ± 4.41 s, 15.5 ± 1.56 s vs 28.67 ± 5.71 s, 21.71 ± 3.4 s, P < 0.05), and the PT in the HBAL group was shortest of all the groups. The albumin in the BAL and HBAL groups significantly increased and a significantly higher level was observed in the HBAL group compared with the BAL group (27.7 ± 1.7 g/L vs 25.24 ± 1.93 g/L). In the HBAL group, the ammonia levels significantly decreased from 54.37 ± 6.86 to 37.75 ± 6.09 after treatment (P < 0.05); there were significant difference in ammonia levels between other the groups (P < 0.05). The levels of antibodies were similar before and after treatment. The PERV RNA and the RT activity in the canine plasma were all negative. CONCLUSION: The HBAL showed great efficiency and safety in the treatment of acute liver failure.

Hepatitis B virus infection and replication in primarily cultured human fetal hepatocytes

AIM: To investigate the infection and replication of hepatitis B virus (HBV) in primarily cultured human fetal hepatocytes (HFHs). METHODS: The human fetal hepatocytes were cultured in serum-free medium, HBV-positive serum was added into the medium to study the susceptibility of hepatocytes to HBV infection. The supernatant was collected for ELISA assay of HBsAg and HBeAg, and quantitative fluorescence PCR for HBV-DNA assay daily. Albumin and HBcAg, CK8 and CK18 expressions were detected by immunohistochemistry in cultured hepatocytes. Content of lactate dehydrogenate (LDH) was measured to find out the integrity of the cell membrane. RESULTS: A stable hepatocyte culture system was established. HBV could infect the hepatocytes and replicate, and HBcAg expression could be detected by immunohistochemistry in hepatocyte-like cells. HBV- DNA in the supernatant could be detected from d 2 to d 18 and HBsAg and HBeAg were positive on d 3-d 18 after HBV infection. HBV in medium increased from d 0 to d 6 and subsequently decreased as the cells were progressively loosing their hepatocyte phenotypes. CONCLUSION: HBV could infect human fetal hepato- cytes and replicate. This in vitro model allowed a detailed study on early events associated with human HBV entry into cells and subsequent replication.

Update on autoimmune hepatitis

Autoimmune hepatitis (AIH) is a necroinflammatory liver disease of unknown etiology that occurs in children and adults of all ages. Characteristics are its autoimmune features,hyperglobulinemia (IgG),and the presence of circulating autoantibodies,as well as a response to immunosuppressant drugs. Current treatment consists of prednisone and azathioprine and in most patients this disease has become very treatable. Over the past 2 years,a couple of new insights into the genetic aspects,clinical course and treatment of AIH have been reported,which will be the focus of this review. In particular,we concentrate on genome-wide microsatellite analysis,a novel mouse model of AIH,the evaluation of a large AIH cohort for overlap syndromes,suggested novel criteria for the diagnosis of AIH,and the latest studies on treatment of AIH with budenoside and mycophenolate mofetil.

Variable expression of cystatin C in cultured trans-differentiating rat hepatic stellate cells

AIM. To study the expression of cystatin C （CysC）, its regulation by transforming growth factor-β1 （TGF-β1） and platelet-derived growth factor （PDGF） and the potential interference of CysC with TGF-β1 signaling in this special cell type. METHODS： We evaluated the CysC expression in cultured, profibrogenic hepatic stellate cells and transdifferentiated myofibroblasts by Northern and Western blotting and confocal laser scanning microscopy. RESULTS： CysC was increased significantly in the course of trans-differentiation. Both TGF-β1 and PDGFBB suppressed CysC expression. Furthermore, CysC secretion was induced by the treatment with TGF-β1. Although CysC induced an increased binding affinity of TGF-β receptor type Ⅲ （beta-glycan） as assessed by chemical cross-linking with [^125I]-TGF-β1, it did not modulate TGF-β1 signal transduction as shown by evaluating the Smad2/3 phosphorylation status and [CAGA]-MLP-luciferase reporter gene assay. Interestingly, the shedding of type Ⅲ TGF-β receptor beta-glycan was reduced in CysC-treated cells. Our data indicated that CysC expression was upregulated during transdifferentiation. CONCLUSION： Increased CysC levels in the serum of patients suffering from liver diseases are at least partially due to a higher expression in activated hepatic stellate cells. Furthermore, TGF-β1 influences the secretion of CysC, highlighting a potentially important role of cysteine proteases in the progression of hepatic fibrogenesis.

Artificial neural network in studying factors of hepatic cancer recurrence after hepatectomy

Objective: To explore the affecting factors of liver cancer recurrence after hepatectomy. Methods:The BP artificial neural network - Cox regression was introduced to analyze the factors of recurrence in1 457 patients. Results: The affecting factors statistically significant to liver cancer prognosis was selected.There were 18 factors to be selected by uni-factor analysis, and 9 factors to be selected by multi-factor analysis. Conclusion: The 9 factors selected can be used as important indexes to evaluate the recurrence of liver cancer after hepatectomy. The artificial neural network is a better method to analyze the clinical data, which provides scientific and objective data for evaluating prognosis of liver cancer.

Perioperative insulin therapy using a closed-loop artificial endocrine pancreas after hepatic resection

Postoperative hyperglycemia is common in critically ill patients, even in those without a prior history of diabetes mellitus. It is well known that hyperglycemia induced by surgical stress often results in dysregulation of liver metabolism and immune function, impairing postoperative recovery. Current evidence suggests that maintaining normoglycemia postoperatively improves surgical outcome and reduces the mortality and morbidity of critically ill patients. On the basis of these observations, several large randomized controlled studies were designed to evaluate the benefit of postoperative tight glycemic control with intensive insulin therapy. However, intensive insulin therapy carries the risk of hypoglycemia, which is linked to serious neurological events. Recently, we demonstrated that perioperative tight glycemic control in surgical patients could be achieved safely using a closed-loop glycemic control system and that this decreased both the incidence of infection at the site of the surgical incision, without the appearance of hypoglycemia, and actual hospital costs. Here, we review the benefits and requirements of perioperative intensive insulin therapy using a dosed-loop artificial endocrine pancreas system in hepatectomized patients. This novel intensive insulin therapy is safe and effectively improves surgical outcome after hepatic resection.

Treatment of hepatic failure with artificial liver support system

Objective To assess the effectiveness of artificial liver support system (ALSS) treatment in patients with hepatic failure. Methods 235 cases of hepatic failure were treated with ALSS in our hospital. All data were analyzed by SPSS. The effectiveness of ALSS treatment was compared according to different stages (i.e., early, middle and end stages). Results 108 patients survived after therapy of ALSS. After each ALSS treatment, the liver function of these patients was greatly improved, the serum endotoxin and HBV-DNA concentrations were significantly decreased, and the serum concentration of aromatic amino acids (AAA) such as methionine decreased while the ratio of branched chain amino acids and aromatic amino acids (BCAA/AAA ratio) increased; patients treated with ALSS in the early or middle stages of disease had much higher survival rates than patients in the end stage of disease.Conclusion ALSS is a reliable therapy for advanced liver diseases and treatment at early or middle stages is appropriate.

Chimera formation of platelet GPⅡb Bak a/b by intrauterine transplantation of fetal liver stem cells

Abstract:Objective To investigate whether artificial heterozygous chimeras of platelets can be established by intrauterine transplantation of fetal liver stem cells and evaluate its potential use for the treatment of Glanzmann thrombasthenia.Methods Platelet glycoprotein (GP) Ⅱb Bak a/b (or GPⅡb Ⅰle843Ser) was used as a genetic marker. A homozygous 16-week-old Bak a/a fetus (as donor) and a homozygous 16.5-week-old Bak b/b fetus (as recipient) were screened from 42 pregnant women hospitalized for abortion. PCR with allele specific primers and FOK Ⅰ digestion based on PCR products were used. Aborted donor fetal liver cell suspensions were prepared and intrauterine transplantation was carried out by infusion of 4?ml fetal liver cells (22×105) into the recipient umbilical vein under ultrasonic visualization.Results At gestation termination (abortion), 21 days after transplantation, chimera GPⅡb Bak a/b of the recipient were detected by FOK 1 digestion based on PCR from DNA and RT-PCR from platelet RNA. Conclusion Intrauterine transplantation of fetal liver cell may provide an effective way for curing GT or other inherited diseases.

Isolation and cultivation of porcine hepatocytes for extracorporeal artificial liver support system

Objective To develop procedures for the successful harvesting of large quantities of viable and functional pig liver cells from abattoir organs.Methods The procedure included partial liver lobe retrograde perfusion and mechanical/enzymatic digestion of the liver tissue, followed by separation of the hepatocytes, based on size and density, from contaminating cell types.Results Digestion of the partial liver lobe resulted in an average yield of 1.39×109 cells (9.9×106 cells/g liver) with an average viability of 92.5%. The yield and viability of cells were improved by dispase/collagenase resultant digestion. The emergence of blebby cells was blocked by supplying oxygen to the cell isolation buffers. Isolated hepatocytes seeded onto polystyrene surfaces remained viable and functional at a level comparable to that of rat hepatocytes, although their function decreased over time.Conclusions Adult pig hepatocytes can be harvested with high yields and retain viability and differentiated function using this method. Abattoir pig livers can be an excellent source of hepatocytes for use as the biological component of artificial liver assist devices.