A simplified model of hypoxic injury in primary cultured rat hepatocytes.

The Anaeropack system for cell culture, which was originally designed for the growth of anaerobic bacteria, was used to produce a hypoxic atmosphere for cultured hepatocytes. We measured changes in the oxygen and carbon dioxide concentrations and the atmospheric temperature in an airtight jar. We also measured changes in the pH of the medium during hypoxia to assess the accuracy of this system. Moreover, we used three durations (2, 3, and 4 h) of hypoxia and 8 h of reoxygenation in cultured rat hepatocytes, and then measured the lactate dehydrogenase (LDH), ketone body concentration (acetoacetate + beta-hydroxybutyrate), and the ketone body ratio (KBR: acetoacetate/beta-hydroxybutyrate) in the medium in order to assess the suitability of this system as a model for reperfusion following liver ischemia. The oxygen concentration dropped to 1% or less within 1 h. The concentration of carbon dioxide rose to about 5% at 30 min after the induction of the hypoxic conditions, and was maintained at this level for 5 h. No effect of the reaction heat produced by the oxygen absorbent in the jar was recognized. The extent of cell injury produced by changing the hypoxic parameters was satisfactorily reflected by the KBR, the ketone body concentration, and the LDH activity released into the medium. Because this model can duplicate the conditions of the hepatocytes during revascularization following ischemic liver, and the Anaeropack system for cell culture is easy to manipulate, it seems suitable for the experimental study of hypoxic injury and revascularization in vitro.

Preoperative assessment for laparoscopic cholecystectomy: feasibility of using spiral computed tomography.

OBJECTIVE: The authors investigated the preoperative feasibility of using spiral computed tomography (SCT) after intravenous infusion cholangiography (IVC-SCT) for laparoscopic cholecystectomy. SUMMARY BACKGROUND DATA: In laparoscopic cholecystectomy, the aberrant or unusual anatomy of the bile duct and severe inflammation or adhesions around the gallbladder sometimes require a conversion to open surgery. METHODS: Laparoscopic cholecystectomies (LC's) were attempted on 440 patients, and preoperative IVC-SCT also was attempted in all of these patients. Using this spiral scanning technique, the bile ducts, cystic duct, and gallbladder were assessed for contour abnormalities, relative position, and filling defects. Forty-seven patients were diagnosed with having stones in their common bile duct or common hepatic duct. RESULTS: Three-hundred eighty-seven patients out of the 440 patients (88.0%) who were subjected to IVC-SCT had the length and course of their cystic duct successfully determined. Anomalous unions of the cystic duct were seen in 59 (15.2%) of 387 patients with respect to the operative findings, and 48 of 440 patients (10.9%) had severe adhesions to Calot's triangle and the surrounding tissues. In these 48 patients, 45 patients (94%) had a nonvisualized cystic duct on IVC-SCT. The preoperative assessment of the feasibility (dense adhesions obscuring Calot's triangle) of using IVC-SCT demonstrated that the sensitivity, specificity, and accuracy were 93%, 98%, and 94%, respectively. Five patients had to be converted to open surgery, and the overall morbidity rates for patients undergoing laparoscopic cholecystectomy was 0.9% (4 of 440). CONCLUSIONS: The most important factor in assessing the feasibility of using laparoscopic cholecystectomy is not the nonvisualized gallbladder, but the nonvisualized cystic duct on IVC-SCT. IVC-SCT may be of benefit to those patients scheduled to undergo laparoscopic cholecystectomy.

Prolonged decreases in plasma nitrate levels at early postoperative phase after hepato-pancreato-biliary surgery.

Nitric oxide (.NO) is known to influence circulatory, neural, immunologic, and metabolic alterations. To evaluate the clinical significance of .NO production under surgical stress, serial measurements of plasma nitrite plus nitrate levels were performed in 45 surgical patients. Group A included 19 patients who underwent major surgery with uneventful postoperative courses. Group B included 18 patients who underwent laparoscopic cholecystectomy. Group C included 8 patients whose surgery was complicated by intra-abdominal abscesses. Eight healthy volunteers served as controls. Plasma nitrate levels were determined with a redox chemiluminescence .NO analyzer and coincided with measurements made by high-performance liquid chromatography (r = 0.868, p < 0.0001, 58 samples). During laparotomy, arterial nitrate levels correlated well with peripheral, portal, and hepatic venous nitrate levels (r = 0.966, 0.938, and 0.949, respectively; p < 0.0001). A significant decrease in nitrate from preoperative levels in groups A (postoperative day (POD) 1 and 3; p < 0.0005) and B (POD 1, p < 0.0001) was observed; nitrate levels in group C did not decrease for 14 days after surgery. Plasma nitrate levels in groups A and B were significantly different (POD 1 through 6, p < 0.05) and at POD 3 were significantly lower in group A (p < 0.005). Plasma nitrate levels measured before and after fasting or food intake were not significantly different. These results suggest that surgical stress leads to a decrease in the end product of .NO in the whole body, and that the greater the surgical stress the longer the duration of decreased .NO production.

Spiral computed tomography scanning after intravenous infusion cholangiography for biliary duct anomalies.

BACKGROUND: Iatrogenic injury of the bile duct during cholecystectomy represents a failure of surgical technique, especially for laparoscopic surgery. Knowledge of the patient's individual ductal anatomy and anomalies preoperatively would be helpful in avoiding such injuries. Therefore, we investigated the anatomy of the biliary duct and any anomalies using spiral computed tomography (SCT) scanning following intravenous infusion cholangiography (IVC-SCT). MATERIALS: Laparoscopic cholecystectomies (LC) were attempted on 437 patients at the Kansai Medical University. Preoperative IVC-SCT and laparoscopic cholangiography were attempted in all of the patients. RESULTS: An overall anomalous union of the cystic duct was seen in 71 (16.2%) out of the 437 patients subjected to IVC-SCT. The following anomalies were observed: right hepatic duct entry in 7 cases (1.6%), parallel low entry in 17 cases (3.9%), posterior spiral entry in 35 cases (8.0%), anterior spiral entry in 7 cases (1.6%), and accessory duct entry in 5 cases (1.1%). The success rate for the LC was 99.5% (435/437). Three patients were switched to open surgery owing to advanced gallbladder cancer and severe adhesions. The success rate for the laparoscopic cholangiography was 97.2% (423 of 435). Intraoperative right hepatic duct injury occurred in only 1 patient with a bile duct anomaly, and it was repaired with laparoscopic T-tube drainage. CONCLUSIONS: The preoperative examination of the biliary tract by IVC-SCT was technically simple, less invasive, and may helpful in avoiding damage to the bile duct, especially in patients with biliary duct anomalies.

Stimulation of liver regeneration and function after partial hepatectomy in cirrhotic rats by continuous infusion of recombinant human hepatocyte growth factor.

BACKGROUND/AIMS: Radical resection is accepted as one of the most curative treatments for hepatocellular carcinoma. However, most patients have coexisting cirrhosis and their liver function is usually impaired. It is therefore important to stimulate the regeneration and function of the remnant cirrhotic liver after hepatectomy. Hepatocyte growth factor is a potent mitogen that has been suggested to play a crucial role in liver regeneration. METHODS: In this study, we performed 45% hepatectomy in rats with cirrhosis induced by thioacetamide, and administered recombinant human hepatocyte growth factor (rhHGF) with dextran sulfate by continuous infusion into the jugular vein with an infusion pump. RESULTS: rhHGF stimulated an increase in the wet weight of the remnant liver compared with untreated control rats. The proliferating cell nuclear antigen labeling index showed that this increase resulted from the stimulation of DNA synthesis. Serum levels of liver enzymes increased after hepatectomy, but returned to the prehepatectomy level more rapidly in rhHGF-treated rats than in controls. rhHGF increased hepatic protein synthesis above prehepatectomy levels and also markedly increased the serum levels of hepatic lipid metabolites. CONCLUSIONS: These results demonstrate that continuous intravenous infusion of rhHGF enhances the growth and function of the remnant liver in rats with cirrhosis after partial hepatectomy. Therefore, rhHGF may be useful after hepatic resection in patients with cirrhosis.

Prophylactic laparoscopic ovarian ablation for premenopausal breast cancer: medical and economic efficacy.

Worldwide analysis showed a highly significant reduction in the annual rates both of recurrence and of death produced by ovarian ablation. We examined the safety, efficacy, and cost-effectiveness of prophylactic laparoscopic oophorectomy for premenopausal breast cancer. Prophylactic laparoscopic oophorectomy was attempted in 15 selected premenopausal patients with breast cancer. After mastectomy, these patients had laparoscopic ovarian ablation as treatment for breast carcinoma with positive estrogen receptors. To evaluate the economic impact of laparoscopic oophorectomy, comparisons were made between patients treated laparoscopically and those treated with conventional surgery, chemotherapy, and hormonal therapy. The average time required for laparoscopic bilateral oophorectomy was 44 min. Oral intake was resumed the next morning, and the patients were discharged 3 days after surgery. No limitation on physical activity and no complications were required during postoperative days. There was no reduction in the overall costs of laparoscopic surgery and conventional surgery in Japan. The cost of one year's supply of tamoxifen was equivalent to the overall cost of surgical oophorectomy. From medical and economic viewpoints, we conclude that laparoscopic ovarian ablation should be considered an alternative to adjuvant chemotherapy in premenopausal women.

Preoperative determination of the surgical procedure for hepatectomy using technetium-99m-galactosyl human serum albumin (99mTc-GSA) liver scintigraphy.

Technetium-99m-diethylenetriaminepentaacetic acidgalactosyl human serum albumin (Tc-GSA) is a new liver scintigraphy agent which binds to the asialoglycoprotein receptors. We evaluated the preoperative assessment for hepatectomy using Tc-GSA liver scintigraphy. Ninety patients with hepatocellular carcinoma were admitted for elective hepatectomy. Tc-GSA scintigraphy was conducted after the intravenous injection of Tc-GSA, and maximal removal rate of Tc-GSA (GSA-Rmax) was calculated using a radiopharmacokinetic model. Measurement of GSA-Rmax, conventional liver function, and 15-minute retention rate of indocyanine green (ICGR15) was carried out preoperatively. The relationships between liver functions, histological activity index (HAI), ICGR15, and GSA-Rmax values were estimated. A significant correlation was obtained between GSA-Rmax and ICGR15 (r = .534, P < .0001). Preoperative discrepancies between GSA-Rmax and ICGR15 values were seen in 15 patients. In these cases, the GSA-Rmax values correlated well with the total HAI scores (r = .595, P < .02), but no significant correlation was seen between the ICGR15 and HAI scores. Two patients died of postoperative liver failure within 2 months of the operation. These two patients were found to have severe discrepancies between their preoperative GSA-Rmax and ICGR15 values. We concluded that GSA-Rmax might be useful for selecting candidates for hepatectomy and that extended hepatectomies (di- and tri-segmentectomy) are high-risk surgical procedures in the case of low GSA-Rmax scores (below 0.35).

Clinical evaluation of closed suction drainage following hepatectomy.

A prospective randomized study on 186 patients was conducted to determine the influence of closed suction drainage (n = 102) versus open drainage (n = 84) on the incidence of postoperative complications after elective hepatic resection. The patients were randomly allocated between the two groups. A total of 60 complications occurred in 31 of the 84 patients (36.9%) given open drainage, while 24 complications occurred in 15 of the 102 patients (14.7%) given closed suction drainage. The incidence of pleural effusion, postoperative ascites, and infected subphrenic collections was significantly lower in the closed suction drainage group than in the open drainage group, at 31% vs 16% (P < 0.05), 19% vs 3% (P < 0.01), and 17% vs 5% (P < 0.05) respectively. However, both groups showed similar rates of subphrenic hematoma and bile collection. These findings indicate that closed suction drainage significantly reduces the incidence of postoperative complications after elective hepatic resection.

Assessing the eligibility for liver transplantation of patients with fulminant hepatitis.

The spleen to liver volume (S/L) ratio of 13 patients with fulminant hepatitis was determined using X-ray computed tomography (CT), and the correlation between S/L ratio and prognosis was evaluated. The S/L ratio of 6 patients who died was greater than 0.16, whereas that of the remaining 7 patients who survived was less than 0.15. These findings suggest that determining the S/L ratio using CT may be useful for assessing the eligibility of patients with fulminant hepatitis for liver transplantation.

Gallbladder cancer with a low junction of the cystic duct or an anomalous pancreaticobiliary junction.

OBJECTIVE: To evaluate conditions similar to those of carcinogenesis of the gallbladder between the gallbladder with a low junction of the cystic duct (LJCD) and an anomalous pancreaticobiliary junction (APBJ). DESIGN: Retrospective and clinicopathological analysis of patients with gallbladder carcinoma. SETTING: First Department of Surgery, Kansai Medical University. PATIENTS: Examination of 47 patients (7 men and 40 women; average age: 67.8 years) with gallbladder carcinoma revealed 7 patients (14.9%; 1 man and 6 women; average age: 67.8 years) with LJCD and 6 patients (12.8%; 6 women; average age: 60.3 years) with APBJ. METHODS: Clinical findings in both groups were compared with those of the 34 patients who remained after exclusion of the data of the above 7 patients with LJCD and 6 patients with APBJ. The data of the three groups were examined by the chi 2 test at the 5% level of significance. RESULTS: Most of the gallbladder cancer patients with LJCD or APBJ had gallstones. The biliary amylase levels determined in the gallbladder of patients with LJCD or APBJ were remarkably high. CONCLUSION: The results indicate that patients with LJCD or APBJ are more likely to develop carcinoma of the gallbladder. The factors responsible for carcinogenesis may be alteration of the bile content due to reflux of pancreatic enzymes through the LJCD or APBJ, and mechanical irritation due to gallstones. Therefore, these pathological conditions in patients with LJCD are similar to those experienced in patients with APBJ.

[Selective thermocoagulation of unresectable malignant tumors using radiofrequency].

Based on our experimental findings on porcine liver, we have been conducting a clinical trial of selective hyperthermia by radiofrequency (RF) capacitive heating with laparotomy for patients with unresectable malignant tumors. In 10 patients with malignant tumors (8 carcinoma of the pancreas, 2 carcinoma of the gallbladder), laparotomy and RF heating were performed after informed consent. The local heat coagulation was produced by heating equipment using 13.56 MHz radiofrequency produced by Omron Corporation, Japan. Four 2-cm electrode needles were placed in the tumor in a square array at intervals of 2.0 cm. Hyperthermia was given for 30 min with a controlled temperature of 50 degrees C in the RF field (2 x 2 x 2 cm3). That of the surrounding area was maintained at less than 40 degrees C. The calculated volume treated by RF ranged between (2 x 2 x 2 cm3) x 1 and (2 x 2 x 2 cm3) x 6. We followed all patients by computed tomographic (CT) scan 2 weeks after coagulation. Tumor markers in the blood were assayed before and 14 days after heating. Follow-up CT scans demonstrated that after the tumor mass had been heterogeneously enhanced, it changed to a homogeneous low-density area in 6 of 10 patients. The levels of tumor markers decreased to lower than the pre-treatment values in 9 of 10 patients. In all patients, the changes in CT scans and/or decrease in the markers were confirmed. Complications such as bleeding or abscess formation were not observed. It was suggested that the selective hyperthermia was safely produced by this equipment. The encouraging results in these patients justify further clinical trials.

Hepatocyte volume as an indicator of hepatic functional reserve in cirrhotic patients with liver tumours.

Using computed tomography (CT), measurements of whole liver volume have been used for the assessment of pre-operative functional reserve in cirrhotics. However, measurements of hepatocyte volume, which exclude stromal fibrous tissue, are considered to more directly reflect hepatic functional reserve. We investigated the relationship between total hepatocyte volume and each of the parameters of conventional liver function. Indocyanine green (ICG) tests and blood analyses for the assessment of liver function were performed prior to surgery in cirrhotic patients with liver tumours. Pre-operative liver volume was determined by integrating images of each liver area obtained by CT. Liver area was measured by an image processing program that traced the profile of the liver image while excluding the tumorous area. Sections of normal tissue stained by the haematoxylin-eosin method, were obtained from the resected liver. Using these sections, a hepatocyte area: whole tissue area ratio was calculated using the image processing program, by tracing the profiles of the hepatocyte nodules. The total volume of hepatocytes was then calculated by multiplying the liver volume by this ratio. The hepatocyte volume per unit bodyweight was significantly correlated with ICG tests and with many other parameters of normal liver function. However, the liver volume per unit bodyweight was correlated only with the plasma ICG disappearance rate and with the blood platelet count. These observations suggest that the functional reserve of the cirrhotic liver is assessed more precisely by hepatocyte volume than by liver volume.

Inflammatory pseudotumor of the liver: report of a case and review of the literature.

Only 56 cases of inflammatory pseudotumor of the liver have been reported in the world literature since its first documentation in 1953. We report herein the case of a 68-year-old man incidentally found to have a lesion in the right lobe of the liver which closely resembled a neoplasm on imaging studies. Thus, partial hepatic resection was performed and histological examination of the resected specimen revealed a diagnosis of inflammatory pseudotumor. Surgical resection is the preferred treatment for inflammatory pseudotumor of the liver, especially in patients for whom a definite histologic diagnosis cannot be made preoperatively or by intraoperative frozen sections. In fact, most of the patients reported in the literature recovered uneventfully after local resection without any postoperative complications, as did our patient.

[Computed tomography with continuous transport and continuous scanner rotation for laparoscopic cholecystectomy].

We investigated the 58 cases of cholecystolithiasis including 8 cases of choledocholithiasis treated with laparoscopic cholecystectomy. All patients received spiral CT scanning with drip infusion cholangiography (DIC-SCT), and 1.6 patients received endoscopic retrograde cholangiography (ERC), and the detection rates of the gallbladder, the bile duct and the cystic duct were compared. The gallbladder could be seen in 86.2% of cases with DIC-SCT and in 63.8% of cases with conventional DIC. The junction between the cystic duct and the common bile duct could be seen in 18 of 58 cases with DIC and in 49 of 58 cases with DIC-SCT. The DIC-SCT showed significantly superior anatomical details compared with images with conventional DIC. A comparison of DIC, ERC and DIC-SCT revealed that the junction between the cystic duct and the common bile duct could be identified in 14 of 16 cases undergoing ERC, in 13 of 16 cases receiving DIC-SCT and in 4 of 16 cases receiving DIC. Significant differences were noted among DIC-SCT, ERC and conventional DIC. We concluded that DIC-SCT is easy, non-invasive and useful for the preoperative assessment of laparoscopic cholecystectomy and also helpful for avoiding damage to the bile duct.

Use of technetium 99m diethylenetriamine-pentaacetic acid-galactosyl-human serum albumin liver scintigraphy in the evaluation of preoperative and postoperative hepatic functional reserve for hepatectomy.

BACKGROUND: Technetium 99m diethylenetriaminepentaacetic acid-galactosyl-human serum albumin (99mTc-GSA) is a new liver scintigraphy agent that binds to the asialoglycoprotein receptors. We evaluated the clinical use of 99mTc-GSA for the perioperative assessment of hepatectomy. METHODS: Thirty-six patients with hepatocellular carcinoma were admitted for elective hepatectomy. 99mTc-GSA scintigraphy was obtained after the intravenous injection of 99mTc-GSA, and a modified receptor index (MRI) was calculated. 99mTc-GSA scintigraphy, conventional liver function, the plasma disappearance rate, and the 15-minute retention rate of indocyanine green (ICGR15) were carried out before operation and every 1 to 3 months after operation. The relationships between several systemic hemodynamic parameters, histologic activity index (HAI), plasma disappearance rate, and ICGR15 or MRI values were estimated. RESULTS: A significant correlation was obtained between the MRI and ICGR15 (r = 0.6231, p < 0.001). Plasma disappearance rates correlated well with systolic volume and left cardiac work, whereas MRI values did not correlate with these systemic hemodynamics. Preoperative discrepancies between the values of MRI and ICGR15 were seen in eight cases. In these cases the MRI values correlated well with HAI scores (p < 0.05) but there was no significant correlation between ICGR15 values and the HAI scores. CONCLUSIONS: These results suggested the use of 99mTc-GSA scintigraphy as a easy and reliable method for determining liver functional reserve.

Three-dimensional reconstruction of the biliary tract using spiral computed tomography.

Spiral computed tomography (SCT) is a recently introduced technique which enables scanning of one large volume with no interscan intervals. Three-dimensional reconstruction using this technique is emerging as an effective means of identifying complex anatomical relationships. Ninety-two patients with cholecystolithiasis including ten with choledocholithiasis were investigated. All underwent SCT after oral and intravenous infusion cholangiography (IVC-SCT) and 22 were also examined by endoscopic retrograde cholangiography (ERC). IVC-SCT showed significantly more anatomical detail than conventional intravenous infusion cholangiography (IVC). The junction between the cystic duct and the common bile duct could be identified before surgery in 19 of 22 patients undergoing ERC, in 18 of 22 receiving IVC-SCT and in five of 22 undergoing IVC. Significant differences were noted between IVC-SCT, ERC and conventional IVC. Three-dimensional surface reconstruction of the biliary tract is useful in the evaluation of anatomical relationships between the cystic duct and the common bile duct (clearly seen in 86 per cent of cases). Three-dimensional reconstruction using SCT scanning is useful for anatomical assessment before laparoscopic cholecystectomy.

Effect of hepatocyte volume on energy status in the cirrhotic rat liver.

To evaluate the effect of reduced hepatocyte volume on liver energy status, the relationship between the morphologically quantified hepatocyte volume and biochemical parameters, and the difference in nuclear density between the parenchyma and stroma were assessed in rat livers with thioacetamide-induced cirrhosis. The ratio of hepatocytes to whole liver tissue, defined as the 'hepatocyte area ratio', and the nuclear densities of the parenchyma and stroma were calculated microscopically with an image analysing system. Adenine nucleotide, protein and DNA contents, and the ornithine carbamoyltransferase activity in the liver were assayed. In the cirrhotic group, adenine nucleotide content, protein content and ornithine carbamoyltransferase activity were positively correlated with the hepatocyte area ratio, whereas DNA content was negatively correlated with this ratio. The adenylate energy charge of the cirrhotic liver was maintained at a constant level regardless of the ratio. Adenine nucleotide content, protein content and ornithine carbamoyltransferase activity per fractional 'hepatocyte area ratio' in cirrhotic livers were significantly lower than in control livers. The nuclear density of the stroma was significantly greater than that of the parenchyma. These results suggest that the lowered energy status in the cirrhotic liver is not caused by reduced hepatocyte volume but rather by impaired hepatocyte metabolism. In addition, the morphological measurement with an image analysing system was found to be useful for evaluating the effects of decreased hepatocyte volume on biochemical derangements in cirrhotic tissue.

Gallbladder cancer with double pancreatic ducts--a case report.

Various forms of anomalous pancreatic ducts may be encountered. We treated a patient with very rare courses of pancreatic ducts associated with gallbladder cancer. The patient, a 65-year-old female, attended our hospital with epigastric pain and general malaise. Abdominal sonography and a CT scan led to the diagnosis of gallbladder cancer. Endoscopic retrograde cholangiopancreatography revealed two main pancreatic ducts that followed a parallel course and joined the end of the common bile duct to form a common duct. The common duct opened at the ampulla of Vater. Thus, duplicated ducts of Wirsung served as the main pancreatic ducts, with no duct of Santorini. The embryonal mechanism and the clinical significance of double pancreatic ducts remains unknown, and our case is very rare.