Examination of lymph and bile composition following experimental liver transplantation in rat

The lymph flow of the thoracic duct, the protein content of the lymph, bile flow and bile composition are studied during the first three hours of the recirculation phase in experimental liver transplantation. It is established that the lymph flow and the protein content of the thoracic duct decrease as a result of the interruption of hepatic lymph flow of a high protein concentration. The change in bile composition and flow is due to the reduced bile acid excretion of the transplanted hepatic tissue.     

Effect of thermal injury in the rat on transfer of IgA protein into bile.

Severe thermal injury is associated with bacterial sepsis; the intestine is considered a likely source of invasive organisms. Because IgA antibody in bile accounts for much of the specific immune defense of the upper intestinal tract in the rat, the effect of thermal injury on the quantity of IgA protein in bile was examined. Sprague-Dawley rats received a 20% to 30% body surface area burn under anesthesia. Eighteen hours later the common bile duct was cannulated and bile was collected for three hours. Total IgA protein in bile decreased 90% after thermal injury. The bile volume, the concentration of bile protein, and free secretory component did not change significantly. Although blood flow to the liver 18 hours after thermal injury was not changed, there was a significant reduction in total IgA concentration in the circulation; both monomeric (m-IgA) and polymeric IgA (p-IgA) were decreased. This finding may explain, in part, the reduced concentration of IgA protein in bile. Although not examined in this study, decreased local hepatic synthesis and/or transport of p-IgA across the hepatocyte may also contribute to the reduced IgA levels in bile.     

Hilar cholangiocarcinoma associated with sarcoid reaction in the regional lymph nodes

A 74-year-old woman was admitted to our hospital with a 2-week history of jaundice. Percutaneous transhepatic cholangioscopy revealed a nodular tumor originating in the upper part of the common hepatic duct, which was invading the confluence of the right and left hepatic ducts. Microscopic examination of biopsy specimens revealed adenocarcinoma. Abdominal ultrasonography and computed tomography demonstrated multiple enlarged lymph nodes around the extrahepatic bile duct and the common hepatic artery. Laparotomy revealed lymph node enlargement in the hepatoduodenal ligament, behind the pancreatic head, and along the common hepatic and left gastric arteries. Extended left hepatic lobectomy, caudate lobectomy, and resection of extrahepatic bile duct with extended lymph node dissection were performed. The histology of permanent specimen revealed no tumor metastasis but a sarcoid reaction in the lymph nodes. The patient is in good health 21 months after the operation, without any evidence of recurrence. This is the first successfully resected case of hilar cholangiocarcinoma associated with sarcoid reaction in the regional lymph nodes.     

Why does somatostatin cause gallstones?

Long-term administration of the somatostatin analogue, octreotide, is complicated by gallstone formation. Somatostatin is known to inhibit hepatic bile secretion and gallbladder emptying. However, the effect of octreotide on gallbladder bile composition remains unknown. Therefore, we tested the hypothesis that octretide would alter hepatic bile composition and cause gallbladder stasis, thereby increasing gallbladder bile solute concentrations. Fourteen control prairie dogs received daily saline injections, whereas 10 animals received 1 micrograms of octreotide subcutaneously three times per day for 5 days. Cholecystectomy and common bile duct cannulation were then performed. Octreotide increased hepatic bile concentrations of bilirubin monoglucuronide (p less than 0.05), total bilirubin (p less than 0.05), and total protein (p less than 0.01). Rsa, an index of gallbladder stasis, was decreased (p less than 0.01) in the octreotide group. Gallbladder bile total calcium (p less than 0.05), bilirubin monoglucuronide (p less than 0.05), total bilirubin (p less than 0.01), total protein (p less than 0.05), and total lipids (p less than 0.05) were increased in the octreotide group. Animals receiving octreotide also had decreased hepatic (p less than 0.05) and gallbladder (p less than 0.001) bile pH. No differences in cholesterol saturation index were observed. These data suggest that in the prairie dog, octreotide (1) alters hepatic bile composition, (2) causes gallbladder stasis, and (3) increases gallbladder bile calcium, bilirubin, protein, lipid, and hydrogen ion concentrations. We conclude that octreotide causes alterations in gallbladder bile composition that increase the likelihood of cholesterol and calcium bilirubinate precipitation.     

Secretin-induced bile secretion, bile acid output and blood supply to the liver in the dog.

Bile secretion and blood flow in the portal vein and the hepatic artery were determined in cholecystectomized anesthetized dogs before and during continuous infusion of varying doses of secretin. Secretin increases bile volume without alteration of bile acid output. Hepatic arterial flow was not altered by any dose. However, high doses of secretin increased portal venous blood flow significantly. It is concluded that secretin action on liver blood flow mirrors mainly superior mesenteric arterial vasodilation and seems to be a rather pharmacologic than a physiologic response. Bile secretion is not influenced by changes of hepatic blood flow in the physiologic range.     

The effects of liver denervation on the regulation of hepatic biliary secretion.

Effects of liver denervation on bile formation were studied in eight dogs prepared with chronic biliary fistulas. The animals were studied in the basal state, after feeding, and during infusion of glucagon 50 ng/kg/min, secretin 2 U/kg/hr, or somatostatin 200 ng/kg/min. After this first set of experiments the animals underwent a total hepatic denervation that consisted of section of the hepatic ligaments and a careful dissection of the portal vein, hepatic artery, and common duct with stripping of all the surrounding connective tissue and topical application of phenol. The above experiments were then repeated. Denervation did not modify bile flow, or bile salts, cholesterol, or phospholipid concentration or output. Biliary response to glucagon and secretin was similar before and after denervation. Somatostatin had an anticholerectic effect in both intact and denervated animals, but significantly reduced bile salt output only in the intact dogs. Feeding had a choleretic effect pre- and postdenervation, and the infusion of somatostatin following feeding decreased bile flow to the same degree before and after denervation. In the intact animals the output of all three biliary lipids was reduced by somatostatin after feeding but they were unaffected by somatostatin after denervation. Moreover, cholesterol and phospholipid outputs were stable after feeding in intact animals, but significantly decreased after denervation. 14C-erythritol clearance studies indicated no change in the canalicular component of bile flow with denervation, except again during somatostatin suppression of feeding. These data indicate that basal bile flow is normal after denervation but that innervation may play an important role in the modulation of responses to somatostatin and more complex stimuli such as feeding.     

Correlation between cellular ATP level and bile excretion in the rat liver

The influence of the cellular level of adenosine triphosphate (ATP) in the liver on bile excretion was studied in rats. In ischemia, the cellular ATP level decreased rapidly--and, concomitantly, bile flow stopped within 5 min. Administration of L-ethionine i.p. to rats reduced the bile flow rate with decrease in the cellular ATP level. The correlation between the bile flow rate and the cellular ATP level was confirmed in a liver perfusion system. On anoxic perfusion, the ATP level and bile flow rate changed in the same manner as in ischemia. The recovery rates of both on reoxygenation decreased with increase in the anoxic perfusion period. During perfusion under oxygenated conditions, decrease in cellular ATP to various levels by infusion of various concentrations of potassium cyanide, an inhibitor of respiration, resulted in corresponding and concomitant suppression of bile excretion. Kinetic analysis of the bile flow rate revealed a Michaelis-Menten-type curve for the cellular ATP level. The apparent Kms for ATP of bile flow rate in L-ethionine-treated rat liver and liver perfused with potassium cyanide were 1.0 and 1.6 mM, and their Vmax values were 4.1 and 2.5 microliter/min/g liver, respectively. The concentrations of main bile components, such as phospholipids, cholesterol, and taurocholate increased, but their total outputs decreased with decrease in the ATP level, and returned to the normal range with recovery of the ATP level. Thus, it was shown experimentally that the extent of hepatic injury can be assessed simply by monitoring the bile flow rate, which reflects the cellular level of ATP.     

Biliary indocyanine green excretion as a predictor of hepatic adenosine triphosphate levels in patients with obstructive jaundice

BACKGROUND: Correlation of the hepatic adenosine triphosphate (ATP) level with indocyanine green (ICG) excretion into bile was examined in patients with obstructive jaundice after the relief of hyperbilirubinemia by preoperative percutaneous transhepatic biliary drainage (PTBD). METHODS: Patients with complete bile duct obstruction, the mean serum total bilirubin concentration being 13.6 +/- 8.5 (SD) mg/dL, underwent PTBD prior to surgery. Within a few days before surgery when the mean serum total bilirubin level decreased to 1.2 mg/dL, ICG (0.5 mg/kg) was intravenously injected, and the whole bile was collected at 1-hour intervals for 5 hours. The ICG concentration in bile, bile flow rate, amount of ICG excreted in bile, and biliary ICG excretion rate as percentage of the injected dose were determined. At the time of surgery, a small liver tissue sample was obtained immediately after laparotomy without any ischemic procedures, and ATP concentrations were determined. Results of hepatic ATP levels were correlated with laboratory and clinical determinations. RESULTS: The bile flow rate was essentially constant during the 5-hour period, the mean value being 21 mL/hour. The ICG concentrations in bile gradually increased, reached the maximal level in 3 hour, and declined thereafter. The biliary ICG excretion rate for 5 hours was 40% +/- 18% of its injected dose. The biliary ICG excretion rate and amount of ICG excreted in bile for 5 hours significantly (P <0.05) correlated with the hepatic ATP level. The decline index of serum bilirubin during PTBD was also correlated with the hepatic ATP level. The serum ICG retention rate, bile flow rate, maximal ICG concentration in bile, and other liver function tests including serum albumin and cholinesterase levels did not correlate with the hepatic ATP level. CONCLUSIONS: Both the amount of and excretion rate of ICG in bile reflect the hepatic ATP level. Determination of biliary ICG excretion contributes to precise evaluation of hepatic energy status before surgery in patients with obstructive jaundice.     

超声对移植肝肝门部淋巴结反应性增生的诊断和随访价值

目的探讨超声对移植肝肝门部淋巴结反应性增生的诊断和随访价值。 方法回顾性分析2006年2月至2016年2月在浙江大学医学院附属第一医院肝移植中心行肝移植术并接受超声常规随访1年以上1 019例受者的超声资料,分析和总结87例符合移植后肝门部淋巴结反应性增生受者的临床资料和超声影像特征。 结果肝移植术后1年,肝门部淋巴结反应性增生发生率为8.54% (87/1 019),超声发现淋巴结肿大时间中位数为移植术后55 d (7～270 d);肝门部淋巴结增大单发57例,多发30例;淋巴结平均长径(24.6±6.2) mm,横径(17.4±2.9) mm;内部回声均匀,彩色血流信号不丰富。伴发门静脉受压2例,轻、中度狭窄;包绕肝门血管生长并伴有肝内胆管轻度扩张3例。肝门部肿大淋巴结平均消失或缩小时间为(82±57) d。 结论对肝门部淋巴结反应性增生超声特征的认识可以减少一部分移植肝穿刺活检。超声可以作为肝移植术后辅助诊断和监测肝门部淋巴结反应性增生的有效影像手段。     

Caffeine prevents cholesterol gallstone formation

Methylxanthines are known to inhibit in vitro gallbladder absorption. Increased gallbladder absorption has been observed during formation of cholesterol gallstones. Therefore we tested the hypothesis that caffeine would inhibit in vivo gallbladder absorption and thus prevent formation of cholesterol gallstones. Sixteen adult male prairie dogs received a control nonlithogenic diet, and 16 were fed a diet containing 1.2% cholesterol. Half of the animals in each group received caffeine in their drinking water. Gallbladder and hepatic bile were examined microscopically and analyzed for biliary lipids and electrolytes. The gallbladder/hepatic bile ratios of bile acids and sodium were calculated as indices of gallbladder absorption. All eight animals receiving the 1.2% cholesterol diet formed cholesterol gallstones, whereas none of the eight animals fed the cholesterol diet plus caffeine formed gallstones. The cholesterol saturation index was similar, however, in both groups. In animals fed a control diet, the administration of caffeine significantly increased hepatic bile flow and decreased the gallbladder/hepatic bile ratio for both bile acids (5.4 +/- 0.9 vs 3.6 +/- 0.3; p less than 0.05) and sodium (1.26 +/- 0.03 vs 1.12 +/- 0.03; p less than 0.01). In animals fed the high-cholesterol diet, caffeine significantly decreased the ratios for both bile acids (9.0 +/- 1.6 vs 5.3 +/- 0.6; p less than 0.05) and sodium (1.37 +/- 0.06 vs 1.21 +/- 0.01; p less than 0.05), lowered gallbladder bile protein levels, normalized gallbladder stasis, and lowered serum cholesterol levels. In summary, caffeine prevented formation of cholesterol gallstones in this experimental model. The effect of caffeine may be the result of alterations in multiple biliary parameters including the inhibition of gallbladder absorption.     

A validated technique for the analysis of biliary bile acid secretion in donor livers prior to transplantation

Many parameters currently used for the pre-transplant assessment of liver allografts, are not reliable enough in predicting the likelihood of early graft dysfunction or non-function. It is generally accepted that bile secretion is a sign of hepatic function post-transplant and that bile flow shows a close linear relationship to the secretion of bile acids (“apparent choleretic activity”). We have studied bile flow, biliary bile acid concentrations and composition and measured apparent choleretic activity from hepatic bile collected with a new technique under controlled conditions at the time of retrieval from 18 donor livers. More than three samples were collected from each of 13 donors and a total of 65 samples of hepatic bile were analysed. Of these, ten showed typical apparent choleretic activity with a positive slope in the regression line analysis (correlation coefficient of 0.9), validating our collection technique. Received: 1 September 1997 Received after revision: 15 December 1997 Accepted: 9 January 1998     

Usefulness of intraoperative fluorescence imaging to evaluate local anatomy in hepatobiliary surgery

BACKGROUND/PURPOSE: One of the major complications encountered in hepatobiliary surgery is the incidence of bile duct and blood vessel injuries. It is sometimes difficult during surgery to evaluate the local anatomy corresponding to hepatic arteries and bile ducts. We investigated the potential utility of an infrared camera system as a tool for evaluating local anatomy during hepatobiliary surgery. METHODS: An infrared camera system was used to detect indocyanine green fluorescence in vitro. We also employed this system for the intraoperative fluorescence imaging of the arteries and biliary system in a pig. Further, we evaluated blood flow in the hepatic artery, portal vein, and liver parenchyma during a human liver transplant and we investigated local anatomy in patients undergoing cholecystectomy. RESULTS: Fluorescence confirmed that indocyanine green was distributed in serum and bile. In the pig study, we confirmed the fluorescence of the biliary system for more than 1 h. In the liver transplant recipient, blood flow in the hepatic artery and portal vein was confirmed around the anastomosis. In most of the patients undergoing cholecystectomy, fluorescence was observed in the gallbladder, cystic and common bile ducts, and hepatic and cystic arteries. CONCLUSIONS: Intraoperative fluorescence imaging in hepatobiliary surgery facilitates better understanding of the anatomy of arteries, the portal vein, and bile ducts.     

Changes of hepatic blood flow and intrahepatic microvascular bed in rats with chronic biliary obstruction]

The effective hepatic blood flow (EHBF) and intrahepatic microvascular bed (IMB) of rats were studied with hydrogen clearance and India ink perfusion 1, 2, 3 weeks after common bile duct ligation (CBDL). The EHBF decreased significantly in all 3 CBDL groups, compared to control (P < 0.0001). There was no significant difference among CBDL groups (P > 0.05). In a given group no significant difference was noted between the blood flow of the left lateral lobe and that of the middle lobe. The IMB was destroyed severely in all CBDL groups. It was concluded that when chronic biliary obstruction developed the EHBF decreased significantly because of shrinkage of IMB resulting from extensive fibrosis of the liver, and necrosis of liver cells.     

The pathomorphology of the liver after unilateral hepatic duct obstruction with laboratory-chemical observations of the course in the dog

In an experimental study the morphological and functional changes of the liver with unilateral hepatic duct obstruction were investigated over a period of 13 months. In 4 series different parts of the liver were excluded of the bile drainage by hepatic duct ligation after cholecystectomy (group I = 25%, group II = 50%, group III 75%, group IV = 100% of the liver, series V = control group was cholecystectomy only. The clinical outcome, biochemical parameters, liver biopsy were examined regularly. Bacteriologic investigation of the bile and hepatic flow measurement were performed at the beginning and at the end of the study. RESULTS: The clinical symptoms were discrete and the biochemical parameters showed a typical course. After 6 weeks, atrophy of the excluded liver with contralateral compensatoric hypertrophy was found. The microscopic correlation was the secondary sclerosing cholangitis (SSC). After 6 weeks, a concentric periductal fibrosis was to be observed in the periportal area. After 12 weeks, bile duct vanishing with persistence of the arteries and veins was found. After 36-48 weeks, biliary cirrhosis and total destruction of the liver parenchyma was found respectively. Simultaneously a chronic disturbance of the hepatic perfusion was seen. It was caused by a perivenous fibrosis of the terminal vein with obliteration of the lumen by endangiitic proliferations and cavernous transformation. The genesis of SSC seemed not be be influenced by bile contamination. The ligated as well as the unligated bile ducts were infected in 20-50% only. There was no difference in the liver specimens with sterile or contaminated bile. The hepatic flow measurement showed a reduction of the portal blood flow and a rise of the arterial flow depending on the amount of the excluded liver. A better understanding of the pathophysiological sequelae of the unilateral hepatic duct obstruction suggests that the drainage by surgical or radiological methods may not invariably be necessary.     

肝外胆管癌中血管内皮生长因子C(VEGF-C)的表达及意义

目的 研究肝外胆管癌中VEGF-C的表达情况及其临床意义。方法 应用免疫组化Envision二步法测定32例肝外胆管癌和7例正常胆管组织VEGF-C的表达。结果 VEGF-C在肝外胆管癌和7例正常胆管组织中阳性表达率分别为53．13％，0％。胆管癌组织与正常胆管组织VEGF-C表达水平之间有显著性差异，且VEGF-C表达与胆管癌的分化程度、肿瘤大小和淋巴结转移有关。结论 VEGF-C在胆管癌的表达可促进胆管癌淋巴结的转移，对判断胆管癌淋巴结转移、评估预后有一定意义。     

Uptake of indocyanine green by hepatocytes under inflow occlusion of the liver

BACKGROUND: It is not clear that hepatic venous backflow actually contributes to hepatic tissue oxygenation under inflow occlusion of the liver. In order to prove that substances delivered via the hepatic vein can be utilized and/or metabolized in hepatocytes during inflow occlusion, hepatic uptake in bile and excretion of indocyanine green (ICG) were investigated in pigs. MATERIALS AND METHODS: Animals were divided into two groups: an inflow occlusion (IO) group (N = 6) and a total hepatic vascular exclusion (THVE) group (N = 3) using a bypass. One milligram of ICG per kilogram body weight was administered at the beginning of blood flow occlusion, the retention rate in the blood (ICG R) measured, and the ICG in the hepatic tissue measured by near-infrared (NIR) spectroscopy. Furthermore, the ICG concentration was measured in bile excreted by intermittent perfusion of the liver. RESULTS: ICG R declined with time in both groups; however, ICG R in the IO group decreased much faster than in the THVE group. There were significant differences between the two groups after 30 min of occlusion (P < 0.05). ICG in the hepatic tissue could be detected as a peak at 805 nm 10 min after ICG injection, and the peak became steeper with time. On the other hand, ICG was not detected at all in the hepatic tissue after 180 min in the THVE group. ICG was excreted in the bile after 60 min under IO and increased with time. On the contrary, ICG was not excreted in the bile at all under THVE. There were significant differences between the two groups after 90 min (P < 0.05). CONCLUSION: These results indicate that ICG can be extracted in hepatocytes and excreted in bile under IO of the liver. Consequently, substances such as oxygen and drugs, which are delivered via the hepatic vein, can be utilized and/or metabolized in hepatocytes under IO.     

Tc-NGA imaging in liver transplantation: preclinical studies

Tc-99m galactosyl-neoglycoalbumin (Tc-NGA) is a new liver-imaging agent which binds to hepatic binding protein (an hepatocyte-specific membrane receptor). This study evaluated the sensitivity of Tc-NGA kinetics and imaging anatomy to pathologic states that are encountered after liver transplantation. Studies were performed in adolescent pigs under control conditions (18 studies), and after orthotopic liver transplantation (nine studies), common bile duct ligation (three studies), hepatic artery ligation (one study), and hepatic resection (two studies). Anatomic and kinetic data were analyzed. Excellent liver images and minimal kinetic changes were noted after common bile duct ligation. Marked imaging defects and major kinetic alterations were observed after hepatic artery ligation and in the presence of preservation injury. Marked depression in hepatic Tc-NGA uptake was observed during acute rejection. Minor alterations in Tc-NGA kinetics were noted after a 25% hepatectomy. These studies indicate that minimal changes in Tc-NGA uptake occur after common bile duct ligation; Tc-NGA uptake is markedly sensitive to hepatic ischemia; decreased Tc-NGA uptake occurs during acute rejection; and hepatic infarcts are demonstrated promptly after preservation injury. Thus Tc-NGA imaging provides a novel means of evaluating hepatic ischemia, hepatic preservation, and hepatic allograft rejection. Tc-NGA imaging may also provide a means of evaluating hepatic regeneration and hepatocyte retrodifferentiation during regeneration.     

Stimulation of bile output by gastrointestinal hormones following portoenterostomy for biliary atresia

For biliary atresia portoenterostomy with externally draining conduit provides a model for quantitation of hepatic excretory function and for assessment of the physiologic response of the intrahepatic biliary system to gastrointestinal hormones in a human hepatopathologic condition. Four patients with biliary atresia were serially evaluated from 2 weeks to 43 months following total bile diverting portoenterostomy. A fifth patient with no bile flow provided a control for these studies. The pattern of Rose Bengal excretion for three patients with a satisfactory clinical course was different from that of a fourth patient with highly variable flow and persistent cholestasis. Marked volume and bicarbonate concentration increases in bile were noted 30 to 45 minutes after secretin infusion but only in the four patients with bile flow. The volume response to glucagon was more diffuse. Bilirubin and bile acid concentrations decreased in the stimulated bile flow periods and hourly outputs of these cholephils were not increased above basal. During two intervals of low bile output, secretin markedly increased bile flow in the patient with persistent cholestasis establishing the patency of the hepatoenteric anastomosis (functional obstruction) in contrast to the lack of secretin response in the control (structural obstruction).     

Cholestatic effect of cyclosporine in the rat. An inhibition of bile acid secretion

Cyclosporine administration in patients with organ transplants may cause cholestasis. In the rat, intraperitoneal administration of cyclosporine, 10 mg/kg, for three weeks did not cause liver function test abnormalities or hepatic histological lesions. However a significant reduction of bile flow and bile acid secretion rates was observed. The fact that reduction of bile flow was related to a decrease of the bile acid-independent flow suggests that cyclosporine-induced cholestasis results from an inhibition of bile acid secretion. Whether this inhibition is caused by the parental molecule or by cyclosporine metabolites needs to be clarified.