Abstract: Objective：from the discovery of a new blood supply of gallbladder carcinoma--- vasculogenic mimicry (VM), insight into diagnosis and treatment of gallbladder cancer. Methods：74 cases of primary gallbladder carcinoma diagnosed at Tongji Hospital from 1994.1 to 2000.12 and their clinical pathological and demographic data were used; VM of gallbladder carcinoma was detected by HE staining and CD31-PAS double staining (Envision method), single- or multiple- factor clinical parameters, Cox model, Kaplan-Meier survival analysis and log-rank test for gallbladder carcinoma with VM (+)/(-) were done. Two or three-dimensional cultures of GBC-SD and SGC-996 cells, invasion, migration, VM formation assay and hemodynamic of gallbladder cancer in vitro and in vivo were carried out. VM-related signal pathways were studied by immunohistochemistry, immunofluorescence, Western blotting and QRT-PCR. Results: VM in 13.5% (10/74) of gallbladder cancers was found, namely VM of gallbladder cancer is the pipeline structures surrounded by gallbladder cancer cells and non vascular endothelial cells, with the PAS positive, red blood cells inside under the microscope; VM (+) gallbladder cancers being mainly adenocarcinoma with highly blood metastasis; VM is an independent prognostic factor for gallbladder cancer patients with VM, and prognosis of gallbladder cancer patients with VM is poor. VM was found in GBC-SD cell 3-D matrices and xenografts, and the formation of VM through the activation of the PI3K/MMPs/Ln-5γ2 signaling pathway. Conclusions: there is a special biological characteristic of VM in highly aggressive gallbladder carcinoma; because VM cannot be detected by conventional pathology, CD31-PAS double staining for specimens were routinely done in the gallbladder cancer patient with highly malignant, easy to hematogenous metastasis and poor prognosis; the presence of VM should be considered for post-operational recurrence after gallbladder cancer radical resection, poor results of adjuvant chemotherapy and anti-angiogenic targeted therapy, "anti vasculogenic mimicry" adjuvant therapy was adopted.