Aim-To assess the effect of insulin-like growth factor II (IGF-II) on proliferation of hepatocellular carcinoma (HCC) cells. Methods-Expression of IGF-II mRNA and protein was detected in 10 archival HCC specimens using in situ hybridisation and inumunohistochemistry, respectively. Expression of the Ki-67 antigen, a proliferation marker, was determined immunohistochemically on the same sec- MethodsTen archival HCCs complicating liver cirrhosis were studied. All 10 patients had died of their disease. Seven tumours were associated with hepatitis C virus (HCV) and three with hepatitis B virus (HBV) infection. All liver samples had been fixed in 10% buffered formalin and embedded in paraffin wax. Serial, 5 gm thick, sections were cut and mounted on 2% 3-aminopropyltriethoxysilane coated slides. Sections from all tumours were stained with haematoxylin and eosin for histopathological examination. IN SITU HYBRIDISATIONThe in situ IGF-II mRNA hybridisation protocols used were as described by Yun et al.3 Briefly, the hybridisation solution contained 50% formamide, 2 x SSC, 0.15 M NaCl, 0.2 mg/ml Escherichia coli tRNA, 1 mg/ml degraded herring sperm DNA, 0.1 mg/ml bovine serum albumin, 10% polyethylene glycol, 0.2 M dithiothreitol, and [35S] CTP labelled antisense or sense RNA probe (40 000 cpm/pl) and was kept at 50°C overnight. The slides were washed prior to RNase treatment. For autoradiography, slides were dipped in nuclear emulsion and exposed for two weeks at 4°C. After development, the slides were counterstained with haematoxylin and eosin. IMMUNOHISTOCHEMISTRYFor IGF-II, dewaxed sections were rehydrated through an alcohol series and overlaid with a 10% non-fat milk solution to block nonspecific protein binding. Mouse monoclonal antibody directed against rat IGF-II (Amano Pharmaceutical Co, Osaka, Japan) was used at a final dilution of 1 in 400 and incubated with the sections overnight at 4°C. Sections were incubated with biotinylated horse anti-mouse IgG (Vector Laboratories, Burlingame, California, USA) and alkaline phosphatase avidin D (Vector Laboratories). Sections were then treated with biotinylated goat anti-avidin D (Vector Laboratories) and again with alkaline phosphatase avidin D. Nitroblue tetrazolium with 5-bromo-4-chloro-3-indolylphosphate (NBT/BCIP: BluGENE kit, BRL, Gaithersburg, Maryland, USA) was used as the immunoreactive alkaline phophatase substrate.Cellular proliferation was assessed in serial sections using the MIB 1 antibody (Immunotech S.A., Marseille, France) directed against the Ki-67 antigen by means of the streptavidin-biotin complex method. Initially, dewaxed sections were autoclaved for 10 minutes at 120°C in a 0.01 M citrate buffer (pH 6.0) to improve antigen retrieval. MIBI was used at a final dilution of 1 in 100. Naphthol-
IntroductionLaparoscopic surgery is a minimally invasive approach with good treatment outcomes and is currently the standard surgery for colorectal cancer in Japan. Mesenteric closure is considered unnecessary in laparoscopic colorectal surgery because it can damage the bowel and blood vessels. However, an internal hernia may develop if the mesentery is not repaired.Case presentationWe report a case of internal hernia in a 61-year-old male of Japanese ethnicity. The patient had advanced sigmoid colon cancer, early-stage transverse colon cancer, and multiple adenomatous polyposis, and underwent laparoscopically-assisted subtotal colectomy. Bowel obstruction developed six days postoperatively and did not improve with conservative treatment. Abdominal computed tomography detected an internal hernia, prompting emergency surgery in which the ileum protruding into the mesenteric defect and an anastomotic stricture were detected. Reanastomosis, mesentery closure, and ileostomy were performed after hernia repair.ConclusionIn this case, open surgery was necessary due to bowel obstruction after laparoscopic colectomy. This outcome indicated that mesenteric closure should have been performed. Thus, the benefits of mesenteric closure require assessment in future cases.
Intersphincteric resection (ISR) has recently been performed for very low rectal cancer, whereas abdominoperineal resection (APR) is typically reserved for cancers extremely close to the anal verge and/or when the depth of tumor invasion is suspected to involve the intersphincteric space. This is because impairment of anal function is considered unavoidable if the external sphincter (ES) is excised. We describe our technique of ISR with ES resection and discuss its outcomes. This surgical technique may offer major clinical advantages to selected patients and should be considered as an alternative to APR, although careful consideration of anal function is required.
Preoperative malnutrition by 48 hours starvation reduced inflammatory cytokine response and cellular immunity, resulting in an increase in hematogenous liver metastasis.
The aim of this study was to elucidate whether fecoflowmetry (FFM) could evaluate more detailed evacuative function than anorectal manometry by comparing between FFM or anorectal manometric findings and the clinical questionnaires and the types of surgical procedure in the patients who received anal-preserving surgery. Fifty-three patients who underwent anal-preserving surgery for low rectal cancer were enrolled. The relationships between FFM or the manometric findings and the clinical questionnaires and the types of procedure of anal-preserving surgery were evaluated. There were significant differences between FFM markers and the clinical questionnaire and the types of the surgical procedure, whereas no significant relationship was observed between the manometric findings and the clinical questionnaire and the types of the surgical procedure. FFM might be feasible and useful for the objective assessment of evacuative function and may be superior to manometry for patients undergoing anal-preserving surgery.
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