Bone morphogenetic proteins (BMPs) induce not only bone formation in vivo but also osteoblast differentiation of mesenchymal cells in vitro. Tumor necrosis factor ␣ (TNF␣) inhibits both osteoblast differentiation and bone formation induced by BMPs. However, the molecular mechanisms of these inhibitions remain unknown. In this study, we found that TNF␣ inhibited the alkaline phosphatase activity and markedly reduced BMP2-and Smad-induced reporter activity in MC3T3-E1 cells. TNF␣ had no effect on the phosphorylation of Smad1, Smad5, and Smad8 or on the nuclear translocation of the Smad1-Smad4 complex. In p65-deficient mouse embryonic fibroblasts, overexpression of p65, a subunit of NF-B, inhibited BMP2-and Smad-induced reporter activity in a dose-dependent manner. Furthermore, this p65-mediated inhibition of BMP2-and Smad-responsive promoter activity was restored after inhibition of NF-B by the overexpression of the dominant negative IB␣. Although TNF␣ failed to affect receptor-dependent formation of the Smad1-Smad4 complex, p65 associated with the complex. Chromatin immunoprecipitation and electrophoresis mobility shift assays revealed that TNF␣ suppressed the DNA binding of Smad proteins to the target gene. Importantly, the specific NF-B inhibitor, BAY11-7082, abolished these phenomena. These results suggest that TNF␣ inhibits BMP signaling by interfering with the DNA binding of Smads through the activation of NF-B.Bone morphogenetic proteins (BMPs) 2 are members of the transforming growth factor ␤ superfamily (TGF-␤) that were originally identified by their ability to induce ectopic bone formation when implanted into muscle tissue (1, 2). BMP signaling is transduced by two types of transmembrane serine-threonine kinase receptor, type I and type II (3, 4). After type II receptors phosphorylate type I receptors in a ligand-dependent fashion, activated type I receptors phosphorylate downstream molecules in the cytoplasm. After BMP type I receptors phosphorylate Smad1, Smad5, and Smad8 (Smad1,5,8), the three Smads form heteromeric complexes with Smad4 and other transcription factors. These complexes translocate into the nucleus and activate the transcription of target genes, including Id1, which encodes an inhibitor of myogenesis (5). This unique and specific ability of BMPs should be useful for the development of bone regeneration. However, BMPs cannot generate enough of a clinical response to be used in bone regeneration (6 -8). One possible reason might be that inflammatory cytokines inhibit bone formation and osteoblast differentiation induced by BMPs. For example, several lines of evidence have shown that tumor necrosis factor (TNF) ␣ inhibits osteoblast differentiation in multiple models, including fetal calvaria, bone marrow stromal cells, and MC3T3-E1 cells (9 -12).TNF␣ is a non-glycosylated protein of 17 kDa, composed of 157 amino acids, that acts as a pleiotropic pro-inflammatory cytokine (13,14). TNF␣ is produced primarily by activated macrophages but is also produced by a variety of other structural c...

Several essential oils possess pharmacological effects. Among the various constituents of essential oils, 1, 8-cineole has been shown to possess pharmacological effects such as anti-bacterial and anti-inflammatory effects. The effect of 1, 8-cineole on human colorectal cancer cells, however, has not reported previously. In this study, we have investigated the anti-proliferative effect of 1, 8-cineole on human colon cancer cell lines HCT116 and RKO by WST-8 and BrdU assays. The cytotoxicity of 1, 8-cineole was investigated by LDH activity and TUNEL staining. The mechanism of apoptosis by 1, 8-cineole was determined by western blot analyses. In in vivo study, RKO cells were injected into the SCID mice and the effect of 1, 8-cineole was investigated. Specific induction of apoptosis, not necrosis, was observed in human colon cancer cell lines HCT116 and RKO by 1, 8-cineole. The treatment with 1, 8-cineole was associated with inactivation of survivin and Akt and activation of p38. These molecules induced cleaved PARP and caspase-3, finally causing apoptosis. In xenotransplanted SCID mice, the 1, 8-cineole group showed significantly inhibited tumor progression compared to the control group. These results indicated 1, 8-cineole suppressed human colorectal cancer proliferation by inducing apoptosis. Based on these studies 1, 8-cineole would be an effective strategy to treat colorectal cancer.

Fatty acid synthase is highly expressed in many types of human cancers. Cerulenin, a natural inhibitor of fatty acid synthase, induced apoptosis in the human colon cancer cell lines HCT116 and RKO. Oxaliplatin also induced cell death in these cell lines. Cerulenin treatment was associated with reduced levels of phosphorylated Akt, activation of p38 and induced caspase-3 cleavage and finally caused apoptosis. Oxaliplatin induced activation of the p53-p21 pathway and p38. In combination with cerulenin and oxaliplatin, activation of the p53-p21 pathway and p38 occurred in a smaller concentration and finally induced caspase-3 cleavage in a smaller concentration of cerulenin and oxaliplatin. In xenotransplanted SCID mice, the cerulenin + oxaliplatin group significantly inhibited tumor progression compared to the control, cerulenin and oxaliplatin groups. Based on these studies, inhibiting fatty acid synthase would be an effective strategy to treat unresectable colorectal cancer tumors in combination with oxaliplatin. Fatty acid synthase inhibitor would be one of the best counterparts of oxaliplatin, which reduces the dose and side-effects of oxaliplatin and would make it possible to endure the chemotherapy over a longer period.

Background We applied a new concept of ''image overlay surgery'' consisting of the integration of virtual reality (VR) and augmented reality (AR) technology, in which dynamic 3D images were superimposed on the patient's actual body surface and evaluated as a reference for surgical navigation in gastrointestinal, hepatobiliary and pancreatic surgery. Methods We carried out seven surgeries, including three cholecystectomies, two gastrectomies and two colectomies. A Macintosh and a DICOM workstation OsiriX were used in the operating room for image analysis. Raw data of the preoperative patient information obtained via MDCT were reconstructed to volume rendering and projected onto the patient's body surface during the surgeries. For accurate registration, OsiriX was first set to reproduce the patient body surface, and the positional coordinates of the umbilicus, left and right nipples, and the inguinal region were fixed as physiological markers on the body surface to reduce the positional error. Results The registration process was non-invasive and markerlesss, and was completed within 5 min. Image overlay navigation was helpful for 3D anatomical understanding of the surgical target in the gastrointestinal, hepatobiliary and pancreatic anatomies. The surgeon was able to minimize movement of the gaze and could utilize the image assistance without interfering with the forceps operation, reducing the gap from the VR. Unexpected organ injury could be avoided in all procedures. In biliary surgery, the projected virtual cholangiogram on the abdominal wall could advance safely with identification of the bile duct. For early gastric and colorectal cancer, the small tumors and blood vessels, which usually could not be found on the gastric serosa by laparoscopic view, were simultaneously detected on the body surface by carbon dioxide-enhanced MDCT. This provided accurate reconstructions of the tumor and involved lymph node, directly linked with optimization of the surgical procedures. Conclusions Our non-invasive markerless registration using physiological markers on the body surface reduced logistical efforts. The image overlay technique is a useful tool when highlighting hidden structures, giving more information.

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