Kazuyoshi Takeshita

Hepatocellular transplantation for metabolic support in experimental acute ischemic liver failure in rats.

The function of transplanted hepatocytes in acute ischemic liver failure was studied in an experimental model using rats. Ischemic liver failure was induced by occlusion of the proximal portal vein and hepatic artery immediately following extracorporeal portofemoral venous bypass. All rats were bred in a closed colony. Rats in Group 1 were untreated and served as controls (n = 10). Rats in Group 2 received an intrasplenic transplant of hepatocytes (1 × 107 cells) 48 h before liver ischemia (n = 10). Serum ammonia and blood glucose were measured before, and 1 h after, ischemia. Serum ammonia was significantly higher than normal (Group 1, 930 μg/dL vs < 110 μg/dL) after liver ischemia. On the other hand, serum ammonia in group 2 also was elevated (355 μg/dL), but significantly less so than in Group 1 (p < 0.001). Blood glucose was lower in both groups after liver ischemia, compared to normal (50-100 mg/dL), but it was higher in Group 2 (30 mg/dL) than in Group 1 (14 mg/dL, p < 0.01). Liver histology 1 h after ischemia showed similar degrees of necrosis in the two groups. Transplanted hepatocytes were viable, and clearly identified in the splenic parenchyma after ischemia. Intrasplenic transplanted hepatocytes provide temporary metabolic support of acute ischemic liver failure in rats, as reflected by enhanced ammonia removal and gluconeogenesis.

Bile salt tauroursodeoxycholic acid modulation of Bax translocation to mitochondria protects the liver from warm ischemia-reperfusion injury in the rat.

BACKGROUND Tauroursodeoxycholic acid (TUDC) is a hydrophilic bile acid that has a cytoprotective effect in primary biliary cirrhosis and primary sclerosing cholangitis. TUDC also protects hepatocytes from hydrophobic bile acid-induced apoptosis. The aim of this study was to determine whether TUDC ameliorates hepatocyte apoptosis during ischemia-reperfusion injury. METHODS We used a rat model of hepatic warm ischemia-reperfusion injury to assess the effects of TUDC. Male Sprague-Dawley rats were subjected to 1 or 2 hr of normothermic ischemia followed by 3 or 6 hr of reperfusion. The treatment group received TUDC (50 mg/kg) by bolus intravenous injection 30 min before initiation of ischemia, whereas the control group received saline only. Blood samples for biochemical analysis were obtained after 6 hr of reperfusion. Liver biopsies for histological assessment were obtained 3 and 6 hr after reperfusion. Hepatocyte apoptosis was determined by terminal dUTP nick-end labeling. The pro-apoptotic protein Bax was quantified at the mRNA and protein level. RESULTS Treatment with TUDC significantly reduced serum transaminase levels. This was associated with a significant amelioration in the levels of hepatocyte apoptosis in the TUDC-treated group compared with control. Furthermore, Western blot analysis of Bax expression in liver tissue indicated that TUDC inhibited the translocation of Bax from the cytosol to the mitochondria. CONCLUSIONS TUDC significantly reduced hepatic injury in this model. The beneficial effects of TUDC upon hepatocyte apoptosis were related to the modulation of Bax protein translocation.

Hypoxic conditions restore lost sensitivity to the growth inhibitory effect of Transforming Growth Factor beta-1 (TGF beta-1) in proliferating rat hepatocytes in vitro

TGF beta-1 is known to be a growth inhibitor of regenerating liver, and an inducer of hepatocyte apoptosis in primary culture. However, hepatocytes can proliferate after partial hepatectomy even at high serum TGF beta-1 concentrations. In this study we used the primary cultures of rat hepatocytes for 10 days to investigate how TGF beta-1 affects proliferating hepatocytes. DNA synthesis peaked on day 8 of culture, and TGF beta-1-induced apoptosis was significantly suppressed on day 8 compared to days 2, 5, and 10. Flow-cytometric analysis revealed that hepatocytes that had incorporated BrdU were resistant to the apoptotic effect of TGF beta-1, and Northern blot analysis showed that TGF beta receptor mRNA was down-regulated on day 8. Hypoxic conditions restores TGF beta receptor mRNA expression and the lost sensitivity of proliferating hepatocyte to TGF beta-1.

Mechanisms of the Formation of Peritoneal Surface Malignancy on Omental Milky Spots from Low Grade Appendiceal Mucinous Carcinoma

Mechanisms of the Formation of Peritoneal Surface Malignancy on Omental Milky Spots from Low Grade Appendiceal Mucinous Carcinoma Purpose: Omental milky spot (OMS) is considered to have an important role in the formation of peritoneal surface malignancy (PSM). However, human OMS and cancer metastasis has not been fully clarified. The present study demonstrates the mechanisms of the formation of metastasis on the omental milky spots (OMS) from the low grade AMC. Methods and materials: To clarify the mechanism of the formation of peritoneal metastasis in low grade appendiceal mucinous carcinoma (AMC), 195 low grade AMC showing peritoneal cancer index (PCI) of ≤28 were studied for the distribution of peritoneal metastasis. Peritoneum was resected from 10 patients, and was prepared as whole-mount extending specimen. The specimens were studied by 5’-nucleotidase and alkaline phosphatase double stain and immunohistologic staining by D2-40, anti-CD31 and anti- Ki-67 monoclonal antibody. Furthermore, peritoneal parts were observed by scanning electron microscopy. Results: Pelvic and subdiaphragmatic peritoneum were involved in 164 (84%) and 143 (73%) patients. Greater omentum was involved in 135 (69%) patients. Under the SEM observation, no typical milky spots were observed on the peritoneum except for the greater omentum. Surface of OMS was covered with cuboidal mesothelial cells. Between the cuboidal mesothelial cells, many stomata were found. After digestion of OMS by 6N KOH, disk-like peritoneal pouch was detected at OMS. Small holes were found on the collagen plate covered on the bottom of the pouch. Below the pouch, initial lymphatic vessels closed to the pouch with stomata. Agglomerated blood capillaries distributed around the initial lymphatics. Metastasis foci were found around the stomata on OMS, but were not detected on the flat mesothelial cells on the greater omentum.. Conclusion: Peritoneal free cancer cells from low grade AMC may be adsorbed at stomata and adhere on the OMS. Then, they proliferate on OMS.

A New Comprehensive Treatment for Peritoneal Metastases Using Cytoreductive Surgery Combined with Hyperthermic Intraperitoneal Chemoperfusion

A game-changing therapy for peritoneal metastasis (PM), now called “comprehensive treatment” was first established in the late 1990s. The treatment consists of aggressive cytoreductive surgery (CRS) combined with perioperative intraperitoneal/systemic chemotherapy. PM is considered as local disease, and the rationale behind the treatment is to remove macroscopic tumors and eradicate residual micrometastasis using perioperative chemotherapy (POC). Comprehensive treatment consists of laparoscopic evaluation of the tumor load, POC, and CRS. POC includes six procedures including laparoscopic hyperthermic intraperitoneal chemotherapy (LHIPEC), neoadjuvant intraperitoneal/systemic chemotherapy (NIPS), hyperthermic intraoperative intraperitoneal chemotherapy (HIPEC), extensive intraoperative peritoneal lavage (EIPL), early postoperative intraperitoneal chemotherapy (EPIC), and late postoperative systemic chemotherapy. In a study of gastric cancer patients with P0/Cy1 status that employed comprehensive treatment, POC was confirmed to eradicate intraperitoneal micrometastasis. For clinical standardization of HIPEC, the concept of the thermal dose should be introduced. One thermal dose is equivalent to 30-min treatment at 43 °C. In gastric cancer, one thermal dose of HIPEC has been shown to reduce the peritoneal cancer index of 3.5, and changed the peritoneal cytology from positive to negative in 70 % of patients.

Ripstein operation for rectal prolapse.

われわれは最近6年間に9例の完全直腸脱に対し, Ripstein手術を施行し, 良好な結果を得ている. 症例は男性3例, 女性6例, 年齢は41歳から79歳, 平均61歳. 直腸脱出を訴えて来院した症例が多く, 病悩期間は2ヵ月～20年と種々で, 脱出直腸の長さは17cmにおよぶものも認めた. その他には便通異常6例, 出血4例, 疼痛3例であり, またほぼ全例に肛門括約筋緊張低下を認めた. Ripstein手術は, 経腹腔的に腹膜翻転部を切離後, 直腸を授動し, メッシュにて仙骨に挙上固定する術式である. 手術成績は, 術後6年を経過した症例もあるが, 再発は1例もなく, 2例に軽度の便通異常を認めるが, 全例に高い満足が得られている.