Mitotic centromere-associated kinesin is a novel marker for prognosis and lymph node metastasis in colorectal cancer.

Mitotic centromere-associated kinesin (MCAK) is a microtubule depolymerase that is essential for proper kinetochore-microtubule attachment during spindle formation. Overexpression of MCAK has been correlated with aggressive forms of carcinoma, resulting in poor prognosis of colorectal cancer. The purpose of this study was to quantify MCAK expression in malignant and benign colorectal tissues and to determine if MCAK expression levels correlate with clinicopathologic factors and prognosis in colorectal cancer patients. Paired colorectal tissue samples from tumours and the corresponding normal tissues were obtained from 120 patients with colorectal cancer who underwent surgical resection. The real-time reverse transcriptase-PCR and immunohistochemistry were used to analyse mRNA and protein expression status with respect to various clinicopathological factors. MCAK expression was higher in colorectal cancer tissue (P<0.01) than in corresponding normal tissue, and this elevated expression level was markedly associated with factors such as lymph node metastasis (P=0.0023), venous invasion (P=0.019), peritoneal dissemination (P=0.021) and Dukes' classification (P=0.0023). Patients with high MCAK mRNA expression also showed a far poorer survival rate than those with low MCAK mRNA expression (P<0.01). Elevated MCAK expression was an independent predictor of overall survival and lymph node metastasis. These data suggest that MCAK expression may serve as a good marker of prognosis and lymph node metastasis in colorectal cancer.

The significance of measuring the serum total bile acids levels during orthotopic liver transplantation.

BACKGROUND/AIMS: Bile acid is confined to the enterohepatic circulation and consists of intestinal absorption and hepatic elimination. We investigated whether measuring the serum total bile acids (TBA) levels was useful for both evaluating the function of the grafted liver and predicting the outcome in porcine orthotopic liver transplantation (OLT). METHODOLOGY: Twenty-two female Yorkshire pigs undergoing OLT were divided into 2 groups as follows: Group A consisted of 11 pigs which survived over 7 days with an uneventful early post-operative course, while Group B consisted of 11 pigs which died within 5 days due to hepatic failure. The serum TBA levels were measured before and after reperfusion in the recipients. RESULTS: Between Groups A and B, no significant difference was observed in the operative backgrounds including the operation time as well as the cold and warm ischemic time. In Group A, the levels of serum TBA rapidly increased during the anhepatic phase, and thereafter promptly decreased after the reperfusion of the grafted liver. A significant difference was observed in the levels of serum TBA before and after reperfusion (p < 0.01), whereas no significant difference was seen in Group B. The delta TBA, which represents the difference in the levels of serum TBA between just prior to reperfusion and 10 min after reperfusion, was 71.8 +/- 43.5 mumol/L in Group A and 20.1 +/- 34.5 mumol/L in Group B, and demonstrated a significant difference between these 2 groups (p < 0.01). On the other hand, no significant differences were seen in the levels of serum AST, ALT, ALP and TB at each time point between Groups A and B. CONCLUSIONS: The level of serum TBA was found to be a more sensitive parameter and also reflected the developing grafted liver function earlier than the conventional parameters for liver function. Moreover, delta TBA thus appeared to be a valuable predictor for the post-operative outcome.

Intraportal administration of low-dose recombinant human hepatocyte growth factor enhances effects of hepatocellular transplantation.

BACKGROUND/AIMS: Although recombinant human hepatocyte growth factor (rhHGF) is a potent mitogen, the dose used for patients is still not clear and must be low to avoid untoward effects. Firstly, the optimal strategy of the dose and route of rhHGF was investigated. Secondly, low-dose rhHGF, which would induce proliferation of transplanted hepatocytes, was explored using Nagase analbuminemic rats. METHODOLOGY: 1) Concentrations of rhHGF in the portal vein were measured after continuous administration of titrated rhHGF through the jugular vein or portal vein. 2) F344 rat hepatocytes (2 x 10(7) cells) were transplanted in the liver of Nagase analbuminemic rats. On the 7th day, the rats were subjected to a low-dose rhHGF treatment. RESULTS: When the rats were given rhHGF in a dose of 50 micrograms/kg/day, the mean concentration in the portal vein (0.8 +/- 0.1 ng/mL) was almost similar to the minimum concentration which stimulated hepatocyte proliferation in vitro. When low-dose rhHGF (50 micrograms/kg/day) was administered directly into the portal vein following hepatocyte transplantation in Nagase analbuminemic rats, the serum levels of albumin were significantly higher than in other groups. It was found that the concentration of rhHGF in the portal vein were 3.1 +/- 0.5 ng/mL with continuous intraportal infusion and 0.8 +/- 0.1 ng/mL with continuous systemic infusion. CONCLUSIONS: It was found that the minimal dose of rhHGF needed to stimulate hepatocyte proliferation was 50 micrograms/kg/day. With rhHGF (50 micrograms/kg/day), continuous intraportal infusion afforded a more favorable outcome in case of proliferation of hepatocytes.

Influence of serum from rats with fulminant hepatic failure on hepatocytes in a bioartificial liver system.

Fulminant hepatic failure (FHF) is a life-threatening condition marked by many excessively increased unmetabolized toxins and growth factors. Recently developed bioartificial liver (BAL) systems containing hepatocytes can be used to treat patients with FHF However, the behavior of these hepatocytes on exposure to FHF serum in vitro remains unclear. In the present study, we used FHF rat models and the sera from these rats (i.e., FHF serum) contained elevated inflammatory cytokines (TNF-alpha, IL-1beta, and IL-6), HGF, and TGF-beta1. In addition, 1x10(8) hepatocytes were harvested from the livers of inbred rats and incubated with microcarrier beads. Four hours later, the hepatocyte-coated beads were inoculated into a hollow-fiber module (=BAL system). FHF serum or normal control serum circulated for 6 hours through the BAL system. Expressions of mRNA for albumin, GST A1, CYP 1A2, OTC and c-fos were investigated by RT-PCR, and PCNA staining was performed before and after perfusion. The expressions of albumin, GST A1, and CYP 1A2 mRNAs were markedly decreased, whereas those of OTC and c-fos were modestly decreased. PCNA positive cells were low and showed no difference between FHF and normal serum-exposed hepatocytes. In conclusion, the exposure of hepatocytes to hypercytokinemia, including inflammatory cytokines and positive and negative growth factors, caused a loss in liver specific functions. This environment also failed to facilitate hepatocyte regeneration.

Efficacy of Nafamostat Mesilate for improving the performance of a bioartificial liver using porcine hepatocytes.

Our bioartificial liver (BAL) consists of porcine hepatocytes attached to beads and plasma perfused through the system. The function of our BAL lasts for approximately 7 hours. The objective of the present study was to investigate the efficacy of Nafamostat Mesilate (NM), a protease inhibitor and potent complement inhibitor, for improving the performance of the BAL. The experimental groups were divided as follows; the NM group (n=7) where the BAL had porcine hepatocytes with 3.8x10(-4) M, of NM, and the control group where the BAL had no NM. Plasma obtained from patients suffering from hepatic failure was perfused through the BAL for 10 hours. The viability of the porcine hepatocytes and the levels of alanine aminotransferase (ALT) in the human plasma were measured during perfusion. After the 10-hour perfusion, another human hepatic failure plasma was perfused for an additional 1 hour and then the function of the BAL was evaluated. After the 10-hour perfusion, the viability of the hepatocytes in the NM group was 51 +/- 7%, whereas that in the control group was rapidly reduced by 35 +/- 5%. Although the levels of ALT in the human plasma in both groups increased with the perfusion time, those in the NM group were significantly lower than those in the control group (p < 0.05). These results suggest that NM prevented damage to the porcine hepatocytes in human hepatic failure plasma as compared to the control group. In the human hepatic failure plasma before perfusion, the partial thrombin time (PT) and the plasma ammonia (NH3) levels were 19.8 +/- 12% and 288 +/- 102 microg/dl, respectively. Fischer's ratios were 0.98 +/- 0.39. Even after the 10-hour perfusion, the BAL in the NM group significantly improved the levels of PT (38 +/- 10%; p < 0.05), NH3 (214 +/- 34 microg/dl; p < 0.05) and Fischer's ratios (1.4 +/- 0.3; p < 0.05). On the other hand, the BAL in the control group did not show any improvement in those parameters. In conclusion, NM was found to help in maintaining the viability of porcine hepatocytes in human hepatic failure plasma, thereby allowing the porcine hepatocyte-based BAL to function much better.

[Treatment of liver cancer: current status and future prospectives].

The most common liver cancers are hepatocellular carcinoma (HCC), cholangiocellular carcinoma (CCC), and metastatic colorectal cancer. In HCC patients, the extent of the surgical resection is limited due to the functional status of the underlying cirrhotic liver. Limited resection, transarterial catheter embolization, ethanol injection, and microwave coagulation have been applied to treat the patients with liver hypofunction; however, the intrahepatic recurrence rate was relatively high in those patients. Therefore, liver transplantation is the only radical treatment to remove HCC and cirrhotic livers with viral infections. Recent advances in anti-viral agents promise to improve the outcome after liver transplantation in patients with HCC. On the other hand, CCC is outside the indications for liver transplantation because of the broad extension of lymph node and nerve plexus. In liver metastasis from colorectal cancer, overall survival is not greatly improved, although arterial chemotherapy reduces mortality related to liver metastasis. Surgical resection including repeated hepatectomy indicates better survival in patients with liver metastases. In the future, both CCC and metastatic liver cancer could be candidates for gene therapy. For the 21 century, a new therapeutic strategy incorporating clinical evidence, molecular biology, and organ replacement needs to be established for the treatment of liver cancer.

Artificial liver: review and Cedars-Sinai experience.

During the past decade, whole organ transplantation has become the only clinically effective method of treating fulminant hepatic failure and chronic liver failure due to specific genetic, hepatocellular, and anatomic defects of liver function. However, wider application of liver transplantation is restricted by shortage of organ donors, high cost, relatively high morbidity, and need for life-long immunosuppression. As a result, investigators have attempted to develop alternative methods to treat liver insufficiency. These ranged from use of plasma exchange to utilization of detoxification columns and extracorporeal devices loaded with various liver tissue preparations. Recently, advances in hepatocyte isolation and culture techniques, improved understanding of hepatocyte-matrix interactions, availability of new biomaterials, improved hollow-fiber technology, and better understanding of flow and mass transport across semipermeable membranes have resulted in the development of a new generation of liver assist devices. Some of these devices, including the one developed by the authors, are currently being tested in the clinical setting. In this paper, the past experience with liver support systems is reviewed, the present status of the field is critically examined, and the results of a phase I clinical trial with the bioartificial liver, utilizing primary porcine hepatocytes, are summarized.

Spontaneous perforated pyometra presenting as pneumoperitoneum.

BACKGROUND: Spontaneous perforated pyometra presenting as pneumoperitoneum is extremely rare. CASE REPORT: An 80-year-old Japanese female with spontaneous perforating pyometra presenting as pneumoperitoneum is reported. The patient came to our institute with severe abdominal pain. Routine abdominal examination showed muscular defense, and plain chest roentgenograms revealed infradiaphragmatic free gas. Subsequent computed tomography also demonstrated pneumoperitoneum. Laparotomy was performed on the basis of a tentative diagnosis of perforation of the gastrointestinal tract but revealed a perforated pyometra. A simple hysterectomy was performed. The histological diagnosis of the surgical specimen was acute endometritis without neoplasm. The present report is the third case of spontaneous perforated pyometra with pneumoperitoneum to date. CONCLUSION: Although uterine disease presenting as pneumoperitoneum is rare in elderly patients with an acute abdomen, the possibility of a perforated pyometra should be considered in the differential diagnosis.