Also, we tried to assess should VEGF be considered in a routine diagnostic workup of children with neuroblastoma.

Maybe these results could help in the planning further follow-up strategies and antiangiogenic therapy trials. Materials and methods Patients and tumour samples Neuroblastoma tissue samples (n = 56) included in this study were retrieved from the archives of the Institute of Pathology Medical School University of Zagreb, Croatia. They were obtained from patients treated at the Children’s Clinical Hospital Zagreb between 1995 and 2008 at the beginning of disease (first biopsy). Clinical staging was classified according to The International Epigenetics Compound Library supplier Neuroblastoma Staging System (INSS) [1, 25]. Histopathological grading was classified according to Shimada System

and Shimada Age-based Pathologic Classification [26, Autophagy Compound Library 27]. All the histological samples underwent a revaluation and new grading (SS). Patients with stage 1, 2 and stage 4s disease (19 patients) were treated with surgery alone, or surgery and moderate-dose chemotherapy. Patients with stage 3 and 4 (37 patients) were treated with surgery combined with intensive, multiagent chemotherapy either with or without radiotherapy and/or metaiodobenzylguanidine (MIBG) therapy. Fourteen patients with advanced disease, and 3 patients with localized disease with N-myc amplification tumour received megatherapy (myeloablative chemotherapy) followed by autologous or allogeneic hematopoietic stem cell transplantation. As hematopoietic stem cell transplantation for our high-risk patients was started in 1999, there were 2 groups of high risk patients, either treated with or without stem cell transplantation (Table 1). Table 1 Patient characteristics Characteristics No. patients Total number 56 Gender      Male 35    Female 21 Age      Median 35.5 months      Range 2 months – 12 years      >18 months

old 36    ≤ 18 months old 20 Histologic subtype      Stroma-rich   Well differentiated 3 Intermixed 10 Focal nodular 3    Stroma-poor   Undifferentiated Cobimetinib clinical trial 30 Differentiating 10 Histology      Favourable 23    Unfavourable 33 Stage      1 3    2 15    3 20    4 17    4s * 1 Treatment      S 3    S/CTH 32    S/CTH/MIBGT 2    S/CTH/RT 2    S/CTH/BMT 14    S/CTH/MIBGT/BMT 2    S/CTH/BMT/RT 1 Survival      Alive 35    Dead 21 Abbreviations: 4s * in infants with small primary tumours and metastatic disease involving the skin, liver, limited infiltration of the bone marrow, and can spontaneously regress; S, surgery; CTH, chemotherapy; MIBGT, metaiodobenzylguanidine therapy; BMT, bone marrow transplant; RT, radiotherapy Immunohistochemistry Immunohistochemical analysis was performed on formalin-fixed paraffin-embedded tumour sections.