Tsuyoshi Kurokawa

Alterations of DNA methylation and histone modifications contribute to gene silencing in hepatocellular carcinomas

Aim:  The aim of the present study was to examine DNA methylation and histone modification changes in hepatocellular carcinomas (HCC).

Hydrogen peroxide induces the production of tumor necrosis factor-α in RAW 264.7 macrophage cells via activation of p38 and stress-activated protein kinase

The effect of hydrogen peroxide (H2O2) on production of tumor necrosis factor (TNF)-α was examined in RAW 264.7 murine macrophage cells. H2O 2 led to production of TNF-α up to 24 h after the treatment, but not nitric oxide in RAW 264.7 cells. H2O2 induced TNF-α production in mouse peritoneal macrophages as well as RAW 264.7 cells. The H2O2induced TNF-α production was prevented by inhibitors of p38 and stress-activated protein kinase (SAPK/JNK), and H2O 2 induced the phosphorylation of p38 and SAPK. Further, H2O 2 significantly augmented the AP-1 activity, but not nuclear factor (NF)-κB activity in RAW 264.7 cells. A high level of intracellular reactive oxygen radicals (ROS) was detected in H2O2-exposed RAW 264.7 cells. Ebselen, a cell permeable antioxidant, prevented the H 2O2-induced TNFα production. H2O2 significantly enhanced lipopolysaccharide (LPS)-induced TNF-α production. Therefore, H 2 O2 was suggested to induce TNF-α production in macrophages via activating p38 and SAPK/JNK as oxidative stress-related signal pathways.

Hand-assisted laparoscopic anatomical left lobectomy using hemihepatic vascular control technique

AbstractWe report a new approach for laparoscopic anatomical left lobectomy. Although laparoscopic limited resection of the liver has been reported, major liver surgery with a laparoscopic approach remains uncommon. Obstacles to routine laparoscopic surgery on the liver are mainly related to difficulty in retraction with current instrumentation, difficulty in assessing safe margins of resection without the use of tactile sense, and the difficulty of safe parenchymal dissection laparoscopically. We introduce a hand-assisted method that can help in resolving the difficulties and pitfalls associated with laparoscopic liver resection, and in making this surgery safer. The hand is the best atraumatic liver retractor in laparoscopic resection and facilitates the use of laparoscopic ultrasonography. Stable hemostasis can be achieved by proper manual application of vascular clips in case of vascular injury. The hemihepatic inflow control technique used in the present case was the en masse occlusion of Glissons sheath of the left hemipedicle at the bifurcation. This technique was used exactly the same as in open surgery. Major vessels such as the left hemipedicle and left hepatic vein were dissected by endovascular cutter. The patient had an uneventful, quick postoperative recovery. This technique allows a minimally invasive anatomical major surgery for liver tumors.

Results of laparoscopic liver resection: retrospective study of 68 patients

BACKGROUND Although an increasing number of reports and publications have dealt with the laparoscopic approach to liver resection, this procedure remains uncommon, and its feasibility, safety and effectiveness are still not established. There are few reports of the advantages of this approach on postoperative recovery. METHODS From December 1997 to March 2007, laparoscopic hepatic resection were performed in 68 patients. RESULTS There were 52 malignant tumors (36 hepatocellular carcinomas, three intrahepatic cholangiocarcinomas, one cystadenocarcinoma, liver metastases from ten colorectal carcinomas and two other organs) and 16 benign lesions among our 68 patients. Fifteen patients with hepatocellular carcinoma had cirrhosis. The mean tumor size was 3.1 +/- 1.8 cm (range 1.0-14.0 cm), and the tumors were located in every liver segment except segment I. Liver resection was anatomical in 17 patients and consisted of a lobectomy in four patients and a lateral segmentectomy in 13 patients. Non-anatomical resections were performed in 51 patients. The operative time was 214 +/- 93 min. Mean blood loss was 393 +/- 564 g. A hand-assisted laparoscopic method or mini-laparotomy method was required in 35 patients (51.4%). Operative complications occurred mainly in our early cases and included three patients (4.4%) with operative bleeding, 2 of whom (2.9%) requiring a conversion to open surgery. Postoperative complications occurred in seven patients (10.0%), and two of then eventually required a re-operation. The mean hospital stay was 17 days. There were no complications in the more recent cases. CONCLUSIONS The laparoscopic approach for liver tumors is feasible, if the indication is carefully selected. The safety of this procedure depends on the surgical experience of the surgeon and team and the availability of the necessary technology.

Laparoscopic left hemihepatectomy combined with right hemicolectomy for liver tumor and hemorrhagic diverticulosis

We report a case of laparoscopic left lobectomy combined with right hemicolectomy for cystic liver tumor and hemorrhagic diverticulosis of the ascending colon. Mobilization of the right hemicolon was performed first with a complete laparoscopic method. Then the surgeons left hand was inserted into the abdominal cavity through a 75-mm incision made in the right upper quadrant, and the hand-assisted method was used for completion after liver resection. After hepatectomy, the right hemicolon was pulled through the hand port incision. The resection and anastomosis were performed extracorporeally to avoid intra abdominal infection. As a hemihepatic inflow control technique, we used the method of en masse occlusion of Glissons sheath of the left hemipedicle at the bifurcation. The hand facilitates proper traction and exposure of the cut surface, and hemostasis can be achieved by proper application of vascular clips or staplers. The patient had an uneventful, rapid postoperative recovery.

Pifithrin-α, a pharmacological inhibitor of p53, downregulates lipopolysaccharide-induced nitric oxide production via impairment of the MyD88-independent pathway.

The effect of pifithrin (PFT)-α, a pharmacological inhibitor of p53, on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 macrophage-like cells was examined. PFT-α inhibited the production of NO but not tumor necrosis factor (TNF)-α in response to LPS. PFT-α inhibited LPS-induced NO production via reduced expression of an inducible NO synthase (iNOS). Moreover, PFT-α inhibited LPS-induced iNOS expression in p53-silenced cells. PFT-α inhibited the production of interferon (IFN)-β, characteristic of the MyD88-independent pathway of LPS signaling, whereas it did not affect the activation of nuclear factor (NF)-κB and mitogen-activated protein kinases in the MyD88-dependent pathway. PFT-α inhibited poly I:C-induced NO production whereas it did not inhibit IFN-β-induced NO production. Further, PFT-α reduced the expression of IFN regulatory factor 3 that leads to the IFN-β production in the MyD88-independent pathway. The most upstream event impaired by PFT-α was the reduced expression of TNF receptor-associated factor (TRAF) 3 in the MyD88-independent pathway. PFT-α also reduced the in vivo expression of iNOS in the livers of mice injected with LPS. Taken together, PFT-α was suggested to inhibit LPS-induced NO production via impairment of the MyD88-independent pathway and attenuated LPS-mediated inflammatory response.

Roles of naofen, a novel WD-repeat-2 protein, in the CCl4-treated livers--a possible relationship to cell proliferation.

Naofen (GenBank accession no. EF613262), a newly found intracellular protein in the WD-repeat-2 protein family, has been cloned as an anti-verotoxin II antibody immunoreactive substance, and the nucleotide- and amino acid-sequences have been clarified. The present study was undertaken to evaluate the roles of naofen especially in carbon tetrachloride (CCl4-induced cirrhosis model of rats, also in partial hepatectomy. Naofen mRNA expressions were observed from the early phases of cirrhosis development and during regenerative phases after partial hepatectomy, more remarkable in the former. Naofen immunoreactive fragments located in the vascular endothelial cells and peri-vascular spaces in normal livers especially in Glissons areas, being strongly stained in the connective tissues 8 weeks after starting CCl4-injections, besides in the cytoplasm of hepatocytes in pseudo-lobules. In contrast, partial hepatectomy caused a small increase of naofen expressions in the whole hepatocytes, and significantly in the endothelial cells of portal veins and hepatic arterioles. Furthermore, in parallel to the degree of naofen mRNA and protein expressions, the rates of double-nuclei cells to total hepatocytes in the Glissons areas increased in both cirrhosis and partial hepatectomy, suggesting a relationship between naofen expression and mitosis. In in-vitro studies with cell lines, vascular endothelial growth factor, a cell proliferation stimulant, increased the naofen mRNA expressions in HepG(2) cell lines, whereas paclitaxel, a cytotoxic anti-cancer drug, diminished them in NRK52E, both concentration-dependently. These results indicated that naofen immunoreactive fragments play an important role in the cell proliferation, relevant for analyzing the regenerative phases during cirrhosis developments and after partial hepatectomy.

Case of cholangiocellular carcinoma in a patient with glycogen storage disease type Ia

Glycogen storage disease (GSD) type Ia is caused by a deficiency in glucose‐6‐phosphatase. Long‐term complications, including renal disease, gout, osteoporosis and pulmonary hypertension, develop in patients with GSD type Ia. In the second or third decade, 22–75% of GSD type Ia patients develop hepatocellular adenoma (HCA). In some of these patients, the HCA evolves into hepatocellular carcinoma. However, little is known about GSD type Ia patients with HCA who develop cholangiocellular carcinoma (CCC). Here, we report for the first time, a patient with GSD type Ia with HCA, in whom intrahepatic CCC was developed.

Necrosis of Large Hepatocellular Carcinoma Induced by Preoperative Portal Vein Embolization: A Case Report

A 71-year-old man consulted our hospital because of a huge liver tumor. He had consumed a considerable amount of alcohol over 40 years. He received medications for hypertension and diabetic mellitus. In the laboratory data, hepatitis virus was negative. Tumor markers AFP and PIVKA-II were elevated to 143,360 ng/ml and 244,000 mAU/ml, respectively. In contrast-enhanced CT, the tumor recognized in the right lobe of liver was heterogeneously enhanced in the early phase and had low density in the late phase (Fig. 1). The preoperative diagnosis was HCC, and right liver lobectomy was planned. The FLR was 265.8 ml and its ratio was 26.6 % according to CT volumetry. Percutaneous transhepatic portal embolization for the right portal vein was performed to increase the FLR. Hepatic arteriography and portgraphy were also performed, and tumor stain was recognized in arteriography (Fig. 2). The left lobe was hypertrophied 3 weeks after PVE, and the