Tatsuya Kin

Ki-ras mutations and p53 protein expressions in intrahepatic cholangiocarcinomas : relation to gross tumor morphology

BACKGROUND & AIMSnWe previously reported that intrahepatic cholangiocarcinomas (ICCs) can be divided into three categories according to their gross appearance with possible links to biological behavior. Ki-ras and p53 gene alterations are thought to be involved in early and late phases of carcinogenesis, respectively. This study was performed to investigate the relationship between the gross appearance and genetic alterations of ICC.nnnMETHODSnWe examined 21 patients with ICC. Ki-ras point mutations were assessed by polymerase chain reaction/single-strand conformation polymorphism methods followed by direct DNA sequencing. Expressions of p53 protein were immunohistochemically assessed.nnnRESULTSnKi-ras point mutations were found in 10 patients (48%), and expressions of p53 protein were detected in 4 (19%). Applying the gross classification that we previously proposed, Ki-ras mutations were prominent in the periductal extension type (4 of 6; 67%) and the spicula-forming type (6 of 10; 60%). On the other hand, none of the five mass-forming-type tumors harbored Ki-ras mutations. Expressions of p53 protein did not show any clear association with gross appearance.nnnCONCLUSIONSnKi-ras gene alterations may be involved in the cholangiocarcinogenesis of periductal extension and spicula-forming but not mass-forming types, suggesting that the underlying processes of development are different.

Repeat Hepatectomy for Recurrent Colorectal Metastases

Abstract. Recurrence rates after hepatic resection in patients with colorectal metastases are reported to range from 47% to 80%. Hepatic recurrence is seen in 35% to 50% of patients. Aggressive surgical resection appears to be a worthwhile treatment in patients with recurrent hepatic metastases to promote longer patient survival because surgical resection remains the only curative therapy available. This is a retrospective review of our experience with 15 patients undergoing repeat hepatic resection culled from 67 patients undergoing initial hepatectomy for metastatic colorectal cancer. Of 67 patients who underwent hepatectomy for colorectal hepatic metastases, 33 developed hepatic recurrence at a median interval of 23 months (range 1–176 months) after the first hepatectomy. The second hepatectomy was performed in 15 patients 5 to 29 months after the first hepatectomy, with no mortality. The mean operating time and blood loss at the second hepatectomy were similar to those at the first hepatectomy. The mean hospital stay at the second hepatectomy was significantly shorter than that at the first hepatectomy. The cumulative survival rate for the 15 patients was 42.4% at 3 years and 21.2% at 5 years, respectively, which compared favorably with the survival rate of the 67 patients who underwent initial hepatectomy. Patients who underwent the second hepatectomy had significantly higher survival rates from the first hepatectomy than the 18 patients with unresectable hepatic recurrence. Repeat hepatectomy can be performed safely and provides long-term survival rates similar to those of first hepatectomies. In appropriately selected patients, repeat hepatectomy for colorectal metastases is a worthwhile treatment.

Transthoracic transdiaphragmatic approach for hepatectomy of Couinaud's segments VII and VIII.

Abstract. For hepatectomy of Couinaud’s segment VII or VIII, severe compression and mobilization of the liver is required to establish the operative field via the usual transabdominal approach. Compression of the cirrhotic liver impairs hepatic and systemic blood circulation, which may cause liver dysfunction. We adopted a transthoracic transdiaphragmatic approach for hepatectomy of segment VII or VIII in cirrhotic patients to establish a good operative field without compressing the liver. The aim of this study was to evaluate the benefits of this approach. Forty-four patients with hepatocellular carcinoma (HCC) complicating liver cirrhosis who underwent limited hepatectomy of Couinaud’s segment VII or VIII were studied. The patients were randomized to two groups preoperatively: group I (n = 22), transabdominal approach; group II (n = 22), transthoracic transdiaphragmatic approach. There were no differences in preoperative liver function tests, hepatic functional reserve, or extent of tumor between the two groups. The operative time in group II was significantly shorter than that in group I (243 ± 50 versus 313 ± 80 minutes;p < 0.01). Operative blood loss in group II was also significantly smaller than that in group I (1190 ± 1098 versus 2679 ± 2267 g;p < 0.01). Serum lactate dehydrogenase levels on postoperative day 1 in group II were significantly lower than those in group I (587 ± 154 versus 791 ± 383 IU/L;p < 0.05). Major postoperative complications were significantly fewer in group II. It was concluded that the transthoracic transdiaphragmatic approach is a useful method for hepatectomy of segments VII and VIII in cirrhotic patients.

Influence of Associated Viral Hepatitis Status on Recurrence of Hepatocellular Carcinoma after Hepatectomy

Abstract. The purpose of this study was to investigate the relation between the recurrence of hepatocellular carcinoma (HCC) and the histologic status of underlying chronic liver disease from a viewpoint of multicentric hepatocarcinogenesis. Sixty-eight patients who underwent curative resection of HCC and have been followed for more than 2 years are reported. Based on the microscopic findings of the noncancerous part of the liver, the patients were divided into normal liver (N,n = 2), chronic persistent hepatitis (CPH,n = 6), chronic aggressive hepatitis (CAH,n = 31), and liver cirrhosis (LC,n = 29) according to a classification by the European Association for the Study of the Liver. Background data for the groups showed no significant differences. Recurrence was observed in none of the patients in the N and CPH groups, 26 (83.9%) of the patients in the CAH group, and 12 (41.4%) of the patients in the LC group. The cumulative disease-free survival rate of the CAH group was significantly lower than that of the CPH group (p < 0.05) and LC group (p < 0.01). This study revealed that the histologic status of the underlying chronic liver disease influenced the recurrence rate in patients with HCC. CAH was considered to be a risk factor for recurrence after resection of HCC.

Humoral Human Xenoreactivity Against Isolated Pig Pancreatic Islets

Abstract It is widely believed that the hyperacute rejection of vascularized xenografts in the pig-to-human combination is triggered by the binding of human preformed natural antibodies (PNAbs) to the Galα.(1,3)Gal epitope in pig endothelium and the subsequent activation of complement. However, it remains poorly defined whether xenogeneic pig pancreatic islets are damaged by antibody and complement-mediated mechanisms. We examined the expression of Galα(1,3)Gal on isolated adult pig islets and the presence of PNAbs in normal human sera directed against islets, using immunofluorescence staining and confocal laser scanning microscopy. The pig islets were not stained with Galα(1,3)Gal-specific lectin GSIB4; however, the exocrine cells reacted strongly with GSIB4, indicating that the Galα(1,3)Gal epitope was highly expressed on exocrine cells, but not on islets. Human sera showed weak reactivity of IgM and IgG class PNAbs to the islets, but strong reactivity to the exocrine cells. Furthermore, we investigated the cytotoxic effect of human serum on pig islets using an in vitro model of pig-to-human islet transplantation. The incubation of pig islets with normal human sera for 45 min resulted in less than 10% specific lysis despite the binding of PNAbs, whereas exposure of porcine aortic endothelial cells to the same human sera caused 56% complement-mediated lysis, determined using a MTT cytotoxic assay. These results support the view that pig islets might not undergo early antibody and complement-mediated rejection in humans.

Effect of complement activation in human serum on isolated porcine islets

We investigated the effect of the activation of complement in human serum on isolated adult porcine islets using an in vitro model of pig-to-human islet transplantation. Pancreata were obtained from slaughterhouse pigs (6-8 mo old). Islets were prepared by intraductal collagenase digestion followed by purification on Ficoll gradients. The purified islets were incubated with xenogeneic human serum with or without heat inactivation for 45 min. As control, islets were incubated with autologous porcine serum. After the incubation, the islets responsiveness to an acute glucose stimulus (5.5 mM, static incubation) was evaluated by measurement of the insulin content of the medium. Islets exposed to human serum showed significantly lower insulin secretory response than the control (1.76 +/- 1.17 microU/islet/120 min, without heat inactivation; 1.74 +/- 1.36 microU/islet/120 min, with heat inactivation; 3.39 +/- 0.92 microU/islet/120 min, control). No difference in insulin secretory response, however, was observed between islets exposed to human serum with heat inactivation and without. Furthermore, we evaluated the cytotoxic activity of human serum on porcine islets by a complement-dependent cytotoxicity using the MTT colorimetric assay, and found that the human serum had no complement-dependent cytotoxic activity against the islets. We concluded that the insulin secretion dysfunction of porcine islets exposed to human serum was not due to the activation of complement and there was no evidence of hyperacute rejection mediated by complement activation in the in vitro model of pig-to-human islet transplantation.

Successful management of portal hypertension following artificial arterioportal shunting: Report of a case

A 63-year-old woman diagnosed as having hepatic hilar cancer underwent an extended left lobectomy of the liver with excision of the right hepatic artery which was involved by the tumor. Because the hepatic artery could not be reconstructed by direct anastomosis, an artificial arterioportal (A–P) shunt was constructed between the common hepatic artery and the portal vein. However, 4 weeks after the operation, portal hypertension with severe esophageal varices developed. Under the diagnosis of portal hypertension caused by excessive blood flow from the A–P shunt, coil embolization of the common hepatic artery was performed using an angiographic technique, following which the esophageal varices completely disappeared. This case demonstrates that portal hypertension after A–P shunting can be effectively treated with coil embolization.

Hyperthermo-Chemo-Hypoxic Isolated Liver Perfusion for Hepatic Metastases: A Possible Adjuvant Approach

As a possible intraoperative adjuvant approach to treating hepatic metastases we developed a method of hyperthermo-chemo-hypoxic isolated liver perfusion in combination with hepatic resection. This method was applied to 11 patients with colorectal hepatic metastases between 1992 and 1995. One patient died on postoperative day 14 of hepatic failure (9% mortality), the cause of which was live temperature that reached 42.9 degree C, which seems to be the maximum limit for thermal toxic effect on the human liver. The other 10 patients tolerated the perfusion well, with mild hepatic and non systemic toxicity after minor or even major hepatic resection; the serum aminotransferase and total bilirubin levels returned to normal levels by postoperative day 14. Only one of eight patients (13%) for whom cytotoxic drugs were added to the perfusate (mitomycin C 10 micrograms/ml or cisplatin 2 micrograms/ml) had hepatic recurrence by 19 months after the perfusion (mean follow-up 25.8 months; median 23 months; range 8-57 months). Two patients were alive with no evidence of disease at 13 and 57 months, respectively after the perfusion; the other five patients had postperfusion extrahepatic recurrences (median: 19 months; range 7-20 months). In contrast, hepatic metastases recurred 7 and 20 months after the perfusion, respectively, in the two patients not given a cytotoxic drug.

Comparison of hemodynamic changes in two veno-venous bypass techniques modified at the portal cannulation site

Veno-venous bypass under total vascular exclusion is a useful technique to permit safer resection of hepatic malignancy. We describe here a retrospective study of two modified venous bypass techniques as alternatives to the conventional end-on portal cannulation technique. Portal decompression via inferior mesenteric vein access was performed in eight patients (group A), and portal decompression via a passive shunt between a branch of the mesenteric vein and the right saphenous vein was performed in a second group (group B; n = 8). Both techniques were used in hepatic resection for malignancy under total vascular exclusion. To assess the efficacy of these bypass techniques, we compared the hemodynamic changes in the two groups. There were no differences in the bypass flow between the two groups. Neither group showed any significant changes in hemodynamic parameters (including mean arterial pressure, cardiac index, systemic vascular resistance index, and pulmonary artery pressure) between the pre-bypass and bypass phases. The heart rate in the bypass phase was significantly increased compared to that in the pre-bypass phase in both groups. All hemodynamic parameters in each phase were similar in the two groups. We conclude that both techniques maintained adequate venous return and stabilized the hemodynamic changes during hepatic resection under total vascular exclusion, and that either technique can be selected according to the intraoperative situation.

Predominant role of local immunosuppressive effect in enhanced efficacy of liposomal FK506 in organ transplantation

Liposomal FK506 is a new formulation of FK506 that increases FK506 levels in the liver and decreases them in the kidney in comparison to conventional IV formulation. In the present study, the efficacy of liposomal FK506 was evaluated in canine kidney and liver transplantation models. Liposomal FK506 increased the immunosuppressive efficacy of FK506 in the liver transplantation model, but decreased it in the kidney transplantation model. These results suggest that local immunosuppressive effects with increased intragraft FK506 level would play an important role in enhancing the immunosuppressive efficacy of liposomal FK506 in liver transplantation.