Records. All information concerning the material included were registered no earlier than 1986. Patient characteristics are summarized inTable 1. The median follow-up was 68 months (range 416 months). No patients have been lost to follow-up. There was no difference within the frequency of hereditary tumor, nonfunctioning tumor, radical C 87 site surgery, poorly differentiated carcinoma, or advanced stage PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19968742 tumor involving the patients included in this study and the bigger unselected material. We did, however, see a tendency toward a longer median survival (P = 0.06) within this patient cohort [10]. Ki-67 Data concerning the proliferation marker Ki-67 were retrieved from patient charts. When information had been not accessible, immunohistochemistry (IHC) for Ki-67 wasWorld J Surg (2012) 36:1411performed at the pathology division laboratory. Paraffin-embedded sections of 4 lm were used for IHC. In 14 situations, no data have been attained. For antigen retrieval, sections have been pretreated with 45 minutes of pressure boiling within a citrate buffer pH 6.0. IHC was performed applying an autostainer (DakoCytomation, Carpinteria, CA, USA). Sections were incubated with an anti-Ki-67 antibody (M7240; DakoCytomation) in antibody diluent (DakoCytomation) at space temperature for 60 min. The reaction product was revealed utilizing Dako kit 50087 (DakoCytomation). Sections were counterstained with Mayer’s hematoxylin. Initial experiments had been performed with omission in the key antibody. All sections were scored by two men and women blinded for patient outcome, as outlined by the percentage of nuclear staining inside the field with the highest percentage of staining (“hot spot”), defined after assessing the whole section (in accordance together with the system applied by Professor Lars Grimelius, Department of Genetics and Pathology, Uppsala). Survivinradiology reports. In ambiguous cases, new pathology assessments were performed. TNM staging Staging was performed in line with the recommended definitions [11]. Stage I was defined as a main tumor confined for the pancreas and \2 cm. Stage IIa was a primary tumor confined for the pancreas and two to 4 cm; stage IIb was a major tumor [4 cm or invading the duodenum or bile duct. Stage IIIa was defined as a tumor invading adjacent organs (stomach, spleen, colon, adrenal gland) or the wall of massive vessels (celiac axis, superior mesenteric artery). The presence of lymph node metastases defined stage IIIb and distant metastases stage IV. Staging reflected the tumor burden at diagnosis and was depending on information retrieved from healthcare records. All cases had been reevaluated determined by existing pathology, surgery, and radiology reports. In ambiguous cases, new pathology assessments had been performed. Statistical analysisSections had been deparaffinized and pretreated in TRS buffer, pH six.0 (S1699; DakoCytomation), inside a stress cooker (Biocare Medical, Concorde, CA, USA). The staining procedure was performed in an autostainer (Autostainer Plus; DakoCytomation). A mouse monoclonal anti-survivin antibody (sc-17779; Santa Cruz Biotechnology, Santa Cruz, CA, USA) was diluted to 1:50 in antibody diluent (DakoCytomation) and incubated with sections for 1 h at area temperature. A Dako EnVision kit (K5007; DakoCytomation) was applied in accordance with the manufacturer’s directions, plus the chromogen 3,30 -diaminobenzidine was applied to reveal the complex. Mayer’s hematoxylin was applied for counterstaining. The key antibody was omitted as the damaging manage in initial experiments; and human tonsil was includ.