Development and reversal of endotoxemia and endotoxin-related death in obstructive jaundice

Gut-derived endotoxemia has been implicated in postoperative complications in patients with jaundice. It is thought that absence of bile in the gut predisposes to portal absorption of endotoxin and endotoxemia is reversed by oral bile salt replacement or internal biliary drainage and return of bile to the gut, but not by external drainage. We believe that the importance of gastrointestinal bile flow has been overestimated and biliary obstruction and the integrity of hepatocyte and Kupffer cell function are more important in the development and reversal of endotoxemia. In experiment 1, serum endotoxin concentrations were measured in control rats (n = 10) after choledochovesical fistula (n = 15) and bile duct ligation (n = 15) and after relief of biliary obstruction by internal drainage (choledochoduodenostomy; n = 8) and sterile external drainage (choledochovesical fistula; n = 8), with a quantitative limulus assay. In experiment 2, mortality rates were measured in similar groups 48 hours after administration of oral endotoxin (5 mg/100 gm) and intravenous lead acetate (5 mg/100 gm). Bilirubin levels were elevated in bile duct ligation (192 +/- 13 mumols/L) compared with control animals and those with choledochovesical fistula, internal drainage, and external drainage (10.6 +/- 1.5 mumols/L). In experiment 1, significant portal endotoxemia and systemic endotoxemia occurred in bile duct ligation (portal, 130.4 +/- 12.9 pg/ml; systemic, 91.8 +/- 11.0 pg/ml) but not in choledochovesical fistula (portal, 49.3 +/- 17.1 pg/ml; systemic, 27.2 +/- 11.5 pg/ml). Relief of obstruction by both internal and external drainage reversed endotoxemia. In experiment 2, significant death occurred in bile duct ligation (13 of 15) but not in choledochovesical fistula (3 of 15), and relief of obstruction by both internal and external drainage prevented death. These results confirm that biliary obstruction is a more important factor than is gastrointestinal bile flow in the development and reversal of endotoxemia.     

Surgical anatomy of the hepatic hilar area

It is important to understand the main variations of the biliary and vascular elements inside the plate system for hilar bile duct carcinoma because all variations of these elements occur in this plate system. The plate system consists of the hilar plate, cystic plate, and umbilical plate which cover the extrahepatic vascular system and are fused with the hepatoduodenal ligament. The bile duct and vascular system that penetrate the plate system form Glisson's capsule in the liver, but the caudate branch and the medial segmental branch are exceptions. The bile duct and hepatic artery accompanying the plate system can be exfoliated from the portal vein with numerous lymph ducts and nerves. The bile ducts in the right hepatic lobe are classified into 4 types, and the standard type is present in 53-72% of cases. In the left bile duct, the medial segmental bile duct is connected in the vicinity of the hilar area in 35.5% of cases, and these cases should be treated the same as the caudate lobe in hilar bile duct carcinoma. Generally, there is little main variation of the portal vein (16-26%), but more variation in the hepatic artery (31-33%). During surgery for hilar bile duct carcinoma, it is important to observe the plate system and the many variations of the bile duct and vascular system.     

Patterns of serum alkaline phosphatase activity in unilateral hepatic duct obstruction: a clinical and experimental study

The levels of serum alkaline phosphatase (ALP) were measured in eight patients with bile duct obstruction limited to one lobe of the liver. Although an initial rise of enzyme concentration was documented in every patient, unrelieved biliary obstruction was associated with a gradual return of ALP to normal values. The return to normal levels coincided with the development of atrophy of that part of the liver deprived of its bile drainage. An animal model of experimental selective biliary obstruction supported a causative association between reduction of hepatocyte mass and a decrease in ALP activity. It appears that normal serum ALP levels can be expected with advanced obstructive biliary disease. Suspected lobar or segmental duct obstruction warrants investigation--even if liver function tests are normal.     

Liver transplantation for “mass-forming” sclerosing cholangitis after laparoscopic cholecystectomy

INTRODUCTIONChronic biliary obstruction consequence of a bile duct injury may require liver transplantation (LT) in case of secondary biliary cirrhosis, intractable pruritus or reiterate episodes of cholangitis. “Mass-forming” sclerosing cholangitis leading to secondary portal vein thrombosis and pre-sinusoidal portal hypertension has not been reported so far.PRESENTATION OF CASEWe present the case of a patient who underwent laparoscopic cholecystectomy for Mirizzi syndrome. The persistent bile duct obstruction due to a residual gallstone fragment was treated by a prolonged biliary stenting. Following repeated bouts of cholangitis, a fibrous centrohepatic scar developed, conglobating and obstructing the main branches of the portal vein and of the biliary tree. The patient developed secondary portal vein thrombosis and portal hypertension. After an extensive diagnostic work-up, including surgical exploration to rule out malignancy, the case was successfully managed by liver transplantation.DISCUSSIONMass-forming sclerosis of the bile duct and biliary bifurcation may develop as a consequence of chronic biliary obstruction and prolonged stenting. Secondary portal vein thrombosis and pre-sinusoidal portal hypertension represents an unusual complication, mimicking Klatskin tumor.CONCLUSIONA timely and proper management of post-cholecystectomy complications is of mainstay importance. Early referral to a specialized hepato-biliary center is strongly advised.     

Biliary bacterial infection in liver surgery

Hepatectomy for biliary tract carcinoma with obstructive jaundice is associated with a higher incidence of postoperative septic complications as compared with hepatectomy for hepatocellular carcinoma or metastatic liver cancer. Since most bacteria isolated from septic sites are identical to those found in the preoperative percutaneous transhepatic biliary drainage (PTBD) bile, bacterial colonization in bile appears to be responsible for posthepatectomy septic complications in patients with biliary tract carcinoma. Although it remains unclear how bile becomes contaminated after bile duct obstruction or why preoperative PTBD increases the incidence of biliary infection, bacterial translocation via the portal vein, resulting from loss of integrity of the intestinal mucosa and change in intestinal microflora, may in part account for the mechanisms. Moreover, impaired function of Kupffer cells and altered structure and function of hepatocyte tight junctions might also participate in the development of postoperative bacteremia in such patients. As septic complications and liver failure are profoundly associated with each other, it is important to take all measures before surgery to enhance liver function and to prevent postoperative septic complications.     

胆道梗阻肝细胞膜Na+/K+ATP酶的变化

目的动态观察在胆道梗阻和梗阻缓解过程中,肝细胞膜Na+/K+ATP酶活性变化的规律。方法对130只胆道梗阻大鼠模型的260份样本进行组织化学染色,结合免疫电镜方法,在肝细胞膜上染色定位Na+/K+ATP酶,计算机图像分析测定,半定量比较肝细胞膜Na+/K+ATP酶活性。结果Na+/K+ATP酶广泛存在于肝细胞膜上,但其活性(相对灰度比值)在肝细胞血窦面、肝细胞间面和肝细胞毛细胆管面呈不均匀分布,分别为107.60±16.64、74.40±9.37、90.30±8.04。反映了肝细胞分泌胆汁的正常极性。Na+/K+ATP酶活性在胆道梗阻和梗阻缓解过程中随着胆红素水平的升高或降低而发生变化,并且随着梗阻和缓解的时间延长而明显下降或改善。在梗阻14d时,分别降为84±10、60±7、65±15;在梗阻14 d缓解1 d时,Na+/K+ATP酶活性分别为86±14、62±6、68±19,而在缓解7 d时其活性分别恢复为96±8、65±6、85±7。结论胆道梗阻对肝细胞膜Na+/K+ATP酶活性的影响可能与胆道梗阻时肝细胞代谢胆红素障碍有关。     

肝内胆管手术入路的解剖及临床应用

目的 探讨显露肝内叶、段胆管的手术入路。方法 研究30例成人肝脏标本的肝内叶、段胆管与血管的毗邻关系。结果 左右肝管均位于肝脏脏面门静脉门静脉左右干的前上缘,左内叶、右前叶胆管位于相应门静脉的前内侧。右后叶胆管位于门静脉右面支或右前叶下段支脏面深侧者占73％（22／30）;位于门静脉右后支脏面深侧或后上缘者占80％（24／30）。左外叶胆管位于门静脉矢状部脏面深侧者占93％（28／30）。选择经肝的脏面显露肝门、左右肝管,经肝的膈面显露肝内叶、段胆管相结合的手术入路,治疗复杂性肝内胆管结石并狭窄患者38例,均获成功。结论 经肝的脏面与膈面相结合的手术入路,比较容易显露和切开肝内胆管及其狭窄段、便于取出结石。     

Usefulness of measuring hepatic functional volume using Technetium-99m galactosyl serum albumin scintigraphy in bile duct carcinoma: report of two cases

We report the usefulness of measuring functional liver volume in two patients undergoing hepatectomy. Case 1 involved a 47-year-old man with hepatitis B virus infection. The indocyanine green test retention rate at 15 min (ICGR15) was 14%. Liver uptake ratio (LHL15) by technetium-99 m galactosyl human serum albumin (^99mTc-GSA) liver scintigraphy was 0.91. The patient displayed hilar bile duct carcinoma necessitating right hepatectomy. After preoperative portal vein embolization (PVE), future remnant liver volume became 54% and functional volume by ^99mTc-GSA became 79%. Although the permitted resected liver volume was lower than the liver volume, scheduled hepatectomy was performed following the results of functional liver volume. Case 2 involved a 75-year-old man with diabetes. ICGR15 was 27.4% and LHL15 was 0.87. The patient displayed bile duct carcinoma located in the upper bile duct with biliary obstruction in the right lateral sector. The right hepatectomy was scheduled. After PVE, future remnant volume became 68% and functional volume became 88%. Although ICGR15 was worse as 31%, planned hepatectomy was performed due to the results of functional volume. In the liver with biliary obstruction or portal embolization, functional liver volume is decreased more than morphological volume. Measurement of functional volume provides useful information for deciding operative indication.     

Portal hypertension after bile duct obstruction: effect of bile diversion on portal pressure in the rat.

Biliary obstruction of 14 and 28 days induced in the rat an increase of portal pressure (PP) and wedge hepatic vein pressure (WHVP); the higher these were, the longer was the obstruction. Occurrence of portal hypertension seemed related to portal and periportal fibrosis. Relief of obstruction after 14 days by bilioduodenal anastomosis brought back to normal PP and WHVP. In rats with longer obstruction periods, bilioduodenal anastomosis failed to lower PP and WHPV although biological signs of cholestasis returned to normal levels. These results suggest that portal hypertension may arise very shortly after biliary obstruction in rats and that it may persist in animals with a prolonged biliary obstruction despite an efficient bile drainage. In clinical conditions, such results would favor early treatment of lesions that usually cause prolonged bile duct obstruction, such as postoperative bile duct stenosis.     

Transplantation of hepatocytes: temporary elimination of scavenger cells prevents early loss of grafted cells

Transplants of isolated syngeneic and allogeneic hepatocytes are rapidly disintegrated, irrespective of the site of engraftment; be it spleen, liver, portal vein, peritoneum, or subcutaneous tissue. Host scavenger cells are responsible for this reaction. We designed a method overcoming early disintegration of the grafted hepatocytes. It consisted of administration of anti-asialoGM1 antiserum eliminating natural killer cells; sublethal whole body irradiation; and reconstitution with syngeneic bone marrow cells, ligation of host bile duct, intrasplenic hepatocyte transplantation, and three consecutive partial hepatectomies. Six months after transplantation a glycogen-rich, trabeculae-forming, dividing hepatocytes, situated along strands of newly-formed fibrous tissue and numerous dilated blind bile cannaliculae were observed. There was evidently more bile canaliculae in hosts with ligated bile duct than nonligated controls. This is the first study showing fibrous tissue formed at the site of hepatocyte implantation, and stellate cells are presumably involved in this process.     

Establishment of practical recellularized liver graft for blood perfusion using primary rat hepatocytes and liver sinusoidal endothelial cells

Tissue decellularization produces a three‐dimensional scaffold that can be used to fabricate functional liver grafts following recellularization. Inappropriate cell distribution and clotting during blood perfusion hinder the practical use of recellularized livers. Here we aimed to establish a seeding method for the optimal distribution of parenchymal and endothelial cells, and to evaluate the effect of liver sinusoidal endothelial cells (LSECs) in the decellularized liver. Primary rat hepatocytes and LSECs were seeded into decellularized whole‐liver scaffolds via the biliary duct and portal vein, respectively. Biliary duct seeding provided appropriate hepatocyte distribution into the parenchymal space, and portal vein–seeded LSECs simultaneously lined the portal lumen, thereby maintaining function and morphology. Hepatocytes co‐seeded with LSECs retained their function compared with those seeded alone. Platelet deposition was significantly decreased and hepatocyte viability was maintained in the co‐seeded group after extracorporeal blood perfusion. In conclusion, our seeding method provided optimal cell distribution into the parenchyma and vasculature according to the three‐dimensional structure of the decellularized liver. LSECs maintained hepatic function, and supported hepatocyte viability under blood perfusion in the engineered liver graft owing to their antithrombogenicity. This recellularization procedure could help produce practical liver grafts with blood perfusion.     

Intrahepatic Segmental Portal Vein Thrombosis After Living-Related Donor Liver Transplantation

Background

Intrahepatic segmental portal vein thrombosis after living-related liver transplantation (LRLT) is uncommon. The cause remains unclear.

Methods

After providing written informed consent, 25 recipients receiving LRLT at our institution from January 2011 to September 2013 were enrolled in this study. We performed triphase computerized tomographic (CT) study of the liver graft of each recipient 1 month after LRLT. The patencies of hepatic artery, portal vein, and hepatic vein were evaluated in detail. The triphase CT scans of the liver of each donor before transplantation also were reviewed. Thrombosis of the intrahepatic segmental portal vein was defined as the occlusion site of the portal vein being intrahepatic. Extrahepatic portal vein thrombosis was excluded in this study.

Results

Among the 25 patients, 2 (8%) developed thrombosis of intrahepatic segmental portal vein. One 47-year-old man received LRLT for hepatitis B viral infection–related liver cirrhosis (Child-Pugh class C) with 3 hepatocellular carcinomas (total tumor volume <8 cm). Another 53-year-old man received LRLT for alcoholic liver cirrhosis (Child-Pugh class C). Both had developed progressive jaundice and cholangitis 1 month after surgery. Intrahepatic biliary stricture was found on the follow-up magnetic resonance images. However, liver triphase CT study demonstrated occlusion of intrahepatic portal vein of segment 8 in each patient. Radiologic interventions and balloon dilatation therapy via percutaneous transhepatic biliary drainage route improved the symptoms and signs of cholangitis and obstructive jaundice for both.

Conclusions

Thrombosis of intrahepatic segmental portal vein is not common but is usually associated with complications of intrahepatic bile duct. Early detection is important, and follow-up CT study of liver is suggested.     

Anatomic correlates of bacterial cholangiovenous reflux

The purpose of these studies was to define the pathways by which bacteria pass from bile duct to bloodstream during acute bacterial cholangitis in the rat. The respective roles of biliary obstruction and intrabiliary pressure during the reflux of biliary bacteria were defined by the infusion of bacteria via the bile duct into rats with or without prior bile duct obstruction. As determined by quantitative blood culture analysis, bacterial reflux from bile to blood was enhanced by increased intrabiliary pressure regardless of presence or absence of biliary obstruction. Light microscopic examination of rat liver 48 hours after bile duct obstruction revealed bile ductular proliferation and bile canalicular dilatation. Light microscopic autoradiographs showed aggregates of tritiated thymidine-labeled Escherichia coli outside of interlobular bile ducts in the portal tracts. Transmission electron microscopic examination of rat liver perfused with a bacterial suspension via the common bile duct showed disruption of liver cells and formation of intracellular vacuoles. Bacteria appeared to enter the sinusoidal spaces via these intracellular vacuoles. We conclude that during retrograde biliary infusion (1) increased intrabiliary pressure is the main determinant of increased bacterial reflux into blood; (2) bacteria enter the bloodstream by predominantly intracellular pathways; and (3) prior biliary obstruction is not a significant factor in bacterial reflux from bile to bloodstream.     

The feasibility of in vivo resection of the left lobe of the liver and its use for transplantation

The anatomical possibility of resecting the left lobe of the liver (segments II and III) in living subjects and using it for transplantation was evaluated. A group of 60 cadaveric livers were dissected at autopsy. The vascular and biliary elements of the left lobe were isolated and the lobe was resected and evaluated for possible grafting. The left lobe was 12-28% (mean 19.4%) of the liver mass. An extrahepatic segment of the left hepatic vein was isolated in 95% of specimens. Arterial blood supply to the left lobe consisted of a single artery (92%) or two arteries (8%). A single portal vein segment to the left lobe (type I) was found in 35% livers. Portal vein branches originated from a common orifice (type II, 35%) or separately (type III, 30%) from the left portal vein, and in these instances, preparation of a portal segment necessitated partial section of the left portal vein wall. Biliary drainage was extrahepatic in 56 livers and consisted of a single duct (type I, 78%), or two ducts (type II, 15%). The resected left lobe was evaluated as satisfactory (single hepatic vein and artery, types I or II portal vein, type I bile duct) in 48% of cases, while a less-satisfactory lobe (type III portal vein or type II bile duct) was obtained in 33%. It was found anatomically difficult or impossible to resect the left lobe for possible transplantation in 11 (19%) liver specimens.     

Medical treatment of primary sclerosing cholangitis

Until 1970, primary sclerosing cholangitis (PSC) was considered to be a medical curiosity. With the development of endoscopic cholangiography, PSC is now recognized more frequently and is a common indication for liver transplantation. PSC is usually progressive, leading to cirrhosis, portal hypertension, and liver failure. The manifestations of disease may be clinically similar to those of other causes of bile duct obstruction and must be distinguished from gallstone disease, bile duct carcinoma, primary biliary cirrhosis, and secondary biliary cirrhosis due to bile duct stricture. Medical management of PSC must take into account the likelihood that destroyed bile ducts do not regenerate as hepatocytes do. Hence, PSC should be treated early in its course. The goal of therapy is to prevent further damage and destruction of bile ducts. In this article, we will present relevant data concerning the medical management of primary sclerosing cholangitis. Received for publication on March 8, 1999; accepted on April 5, 1999     

Research review: DNA polymerases as molecular markers of the regenerating capacity of hepatocytes

Hepatocyte regeneration has been widely investigated, with the mitotic index and the incorporation of [³H]thymidine being used as regeneration markers. We focused on the induction of DNA replication enzymes, particularly DNA polymerases (pol) α, δ, and ε. Using rat models, we have shown that the activity of pol α in crude liver extract well represents the regenerating capacity of hepatocytes. Using pol α as an indicator, we analyzed liver regeneration in rat models under various conditions: obstructive jaundice, external or internal biliary drainage, and the obstruction of portal vein branches. It has been revealed that the ligation of the common bile duct alone induces a certain amount of hepatocyte proliferation. It was striking that external biliary drainage suppressed regeneration capacity in cholestatic rat liver after partial hepatectomy. The strong regeneration in nonligated lobes induced by portal branch ligation was similar to the liver regeneration seen after partial hepatectomy with respect to the induction of DNA polymerases. Taken together, the aspects of DNA replication, particularly the induction of DNA polymerases, may contribute to shedding new light on the regeneration of human liver. This work was supported in part by a Grant-in-Aid for General Scientific Research and for Cancer Research from the Ministry of Education, Science and Culture, Japan, and by grants from the Uehara Memorial Foundation     

Variation in the course of vessels in the vicinity of the hepatic port in biliary atresia

Among 114 patients with biliary atresia, the type of obstruction of the extrahepatic bile duct and the course of the hepatic artery, cystic artery, and portal vein were determined and analyzed from the view point of their mutual relationships and in comparison with postoperative bile excretion. As a result, the following findings characteristic of biliary atresia were obtained: (1) The course and location of the extrahepatic bile duct did not deviate greatly from normal conditions, excluding some exceptional cases, but the site of the origin of the cystic artery differed from that in normal cases. (2) The pattern of the course of the vessels varied among different types of obstruction of the extrahepatic bile duct. (3) There was a difference in postoperative bile excretion in relation to the type of obstruction of the extrahepatic bile duct, the course of the vessels, and their combinations.     

门静脉分支结扎后门静脉压力变化与肝再生的关系

目的 探讨90%门静脉分支结扎后大鼠门静脉压力变化与肝再生的关系.方法 45只雄性SD大鼠行90%门静脉分支结扎术,其中5只进行假手术作为对照.观察不同时相点门静脉压力和非结扎侧肝脏质量变化,光学显微镜下观察非结扎侧肝细胞的形态学变化,免疫组织化学方法检测未结扎侧肝细胞的增殖细胞核抗原(PCNA),TUNEL法检测未结扎侧肝细胞的凋亡情况,并进行定最分析.采用Pearson相关分析和t检验分析数据.结果 95%(38/40)的大鼠存活.结扎侧肝叶进行性萎缩,非结扎侧肝叶占全肝质量的比例随时问推移而增加,12 h内增加较缓慢,仅为10.75%;而1～5 d则增加速度明显加快,达到27.57%;7～28 d达到平台期,缓慢增加到32.37%.术前门静脉压力为(9.1±1.8)cm H_2O(1 cm H_2O=0.098 kPa);结扎后立即升高,12 h达到高峰(15.8±2.7)cm H_2O,与术前比较差异有统计学意义(t=6.847,P<0.05);1～28 d由(13.6±2.3)cm H_2O逐渐下降为(9.3±2.0)cm H_2O.术前大鼠PCNA阳性细胞计数为7%±3%,术后12 h至3 d由14%±5%上升至21%±6%,第5天达到高峰为26%±7%,与术前比较差异有统计学意义(t=9.129,P<0.05),随后逐渐恢复正常.TUNEL法检测结果显示,术前大鼠肝脏和术后各时相点大鼠未结扎侧肝脏仅见极少量凋亡细胞.大鼠门静脉压力与非结扎侧肝叶肝细胞PCNA的表达在术后1、3、5 d呈正相关(r=0.913,0.896,0.908,P<0.05),在术后14 d时相点呈负相关(r=-0.926,P<0.05).结论 大鼠90%门静脉分支结扎术后,引起未结扎侧肝细胞的活跃再生,再生后的肝脏可恢复原来的质量;肝再生以肝细胞增殖加速为主,而非肝细胞凋亡减少;门静脉压力变化在肝再生过程中可能发挥重要作用.     

Changes in liver blood flow with development of biliary obstruction in the rat.

As well as causing retention of biliary secretory products, bile duct obstruction is associated with other hepatic and systemic effects which are poorly understood. These latter changes contribute to the increased morbidity and mortality associated with surgery for biliary obstruction. This study examines the changes in liver blood flow occurring after common bile duct (CBD) ligation in the belief that relative hepatic ischaemia may contribute to dysfunction. A new method for measuring liver blood flow (LBF) based on the clearance of 133Xe from the liver following injection into the portal vein is described. With this new development, serial measurements of LBF can be performed in the conscious, unfasted rat. With the development of jaundice, a highly significant reduction in LBF is seen which is first evident 36 hours after CBD ligation. By the third day after ligation, LBF is only 54% of the control value. No fall in LBF is seen in rats subjected to sham ligation. The mechanism for the reduction in LBF is uncertain, but the delayed onset makes any relfex autonomic reaction unlikely.     

彩色多普勒超声在检测正常移植肝脏及其并发症中的应用价值

目的 探讨彩色多普勒超声在检测正常移植肝脏及并发症中的应用价值。方法 用彩色多普勒超声观察41例肝移植患者肝脏动、静脉血流变化及胆管、肝实质声像图改变。主要观察最大血流速度、时间平均血流速度、阻力指数及胆管的内径。结果 41例中17例（41．5％）发生移植物排斥反应，其彩色多普勒显像（CDFI）显示13例（76．4％）门静脉平均血流速度减低；15例（88．25％）肝静脉波幅降低，呈单相锯齿波或负二相波；9例（52．9％）肝动脉阻力指数升高；5例（29．4％）胆管轻度扩张。41例中5例（12．2％）肝动脉血栓形成，主要表现为门静脉周围检测肝动脉血流信号消失。检测中要注意操作技术及一些假阳性和假阴性影响因素。肝移植术后3例（17．3％）早期轻度胆管扩张，2周内恢复。20例（48．8％）胆管扩张明显并逐渐加重，原于胆管狭窄（2例）、胆管结石（15例）及其他（3例）。结论 彩色多普勒显像技术对正常移植肝脏的确定及并发症的检出具有重要价值；对提高术后存活率有重要意义。