BACKGROUND: CpG DNA plays an important role in immune cell function. This study examined whether the temporal control of toll-like receptor (TLR)9 by CpG DNA can regulate the expression of matrix metalloproteinase-9 (MMP-9). MeETHODS AND MATERIALS: Macrophages were cultured in the presence of 10percent FBS. For the various MMP genes analysis, RT-PCR and real-time PCR were performed. In addition, zymography assay performed for the MMP activity. The phosphorylation assay did for the ERK1/2 and NFkappaB activation, and luciferase promoter assay was for the NFkappaB activity. RESULTS: CpG DNA induced the mRNA expression of MMP-2, MMP-9, and MMP-13, but not of MMP-7, MMP-8, and MMP-12, in a time-dependent manner. Especially, the mRNA expression of MMP-9 was strongly induced by CpG DNA using real-time RT-PCR. The TLR9 inhibitor, chloroquine, suppressed CpG DNA-induced MMP-9 expression and its activity. Moreover, CpG DNA induced the phosphorylation of ERK and the inhibition of ERK by U0126 suppressed CpG DNA-induced MMP-9 expression and its activity. CpG DNA stimulated IkappaB-alpha degradation and luciferase activity. In addition, pretreatment of SN-50, the inhibitor of NFkappaB, strongly blocked the CpG DNA-induced MMP-9 expression and activity. CONCLUSION: These observations suggest that CpG DNA may play important roles in the activation of macrophages by regulating the production of MMP-9 via the sequential TLR9-ERK-NFkappaB signaling pathway.
Chloroquine ; DNA ; Luciferases ; Macrophages ; Matrix Metalloproteinase 9* ; Phosphorylation ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; Toll-Like Receptors*

PURPOSE: Taxol (Paclitaxel) is a new generation of chemotherapeutic drug proven to be effective in the treatment of many cancers. In this study, to further demonstrate the differential effect of the tumor suppressor gene, p53, on the Taxol-induced apoptosis in osteogenic sarcoma cell lines, we used p53-defected SaOS2 cells and wild type p53-expressed U2OS cells. MATERIALS AND METHODS: The cell viability was measured by the XTT assay. To examine whether the differential expressions of p53, in U2OS and SaOS2 cells, were associated with Taxol-induced apoptosis, DNA fragmentation assays were performed on both cytosolic and genomic DNA. Since the cleavage of poly (ADP-ribose) polymerase (PARP) is primarily responsible for apoptosis, the cleavage of PARP, and the expression of cyclin B1, polo-like kinase, Bax, Bcl-xL, Bcl-2 in U2OS and SaOS2 cells were compared by Western blot analyses. RESULTS: The cell viability of the p53-defected SaOS2 cells was markedly decreased with Taxol treatment. Whereas, the cell viabilities due to 6-mercaptopurine and adriamycin were no different between the U2OS and SaOS2 cells. Treatment with Taxol induced a ladder- like pattern of DNA fragments, which is a biochemical hallmark of apoptosis, consisting of multiples of approximately 180-200 base pairs, in a dose-dependent manner in the SaOS2 cells, but insignificantly with the U2OS cells. When the cells were treated with Taxol, the 89 kDa cleavage product of PARP clearly appeared as a function of time in the SaOS2 cells, but not in the U2OS cells. The Taxol-induced apoptosis in p53 defected-osteogenic sarcoma cells was associated with the PARP cleavage as a result of the increased activity of caspase 3, and the high expressions of cyclin B1 and PLK. Bax, as a proapoptotic factor, was increased in the SaOS2cells, but the Bcl-xL and Bcl-2 were decreased when the cells were exposed to 10miceoM Taxol. CONCLUSION: From these results, it was concluded that p53-defected SaOS2 cells are much more sensitive to Taxol-induced apoptosis than p53-expressed U2OS cells.
6-Mercaptopurine ; Apoptosis* ; Base Pairing ; Blotting, Western ; Caspase 3 ; Cell Line ; Cell Survival ; Cyclin B1 ; Cytosol ; DNA ; DNA Fragmentation ; Doxorubicin ; Genes, Tumor Suppressor ; Osteosarcoma* ; Paclitaxel ; Phosphotransferases ; Sarcoma

BACKGROUND: Phosphatidic acid (PA), an important second messenger, is involved in inflammation. Notably, cell-cell interactions via adhesion molecules play a central role in inflammation. This thesis show that PA induces expression of intercellular adhesion molecule-1 (ICAM-1) on macrophages and describe the signaling pathways. MATERIALS AND METHODS: Macrophages were cultured in the presence of 10% FBS and assayed cell to cell adhesion using HUVEC. For the gene and protein analysis, RT-PCR, Western blot and flow cytometry were performed. In addition, overexpressed cell lines for dominant negative PKC-delta mutant established and tested their effect on the promoter activity and expression of ICAM-1 protein by PA. RESULTS: PA-activated macrophages significantly increased adhering to human umbilical vein endothelial cell and this adhesion was mediated by ICAM-1. Pretreatment with rottlerin (PKC-delta inhibitor) or expression of a dominant negative PKC-delta mutant, but not Go6976 (classical PKC-alpha inhibitor) and myristoylated PKC-zeta inhibitor, attenuated PA-induced ICAM-1 expression. The p38 mitogen-activated protein kinase (MAPK) inhibitor blocked PA-induced ICAM-1 expression in contrast, ERK upstream inhibitor didn't block ICAM-1. CONCLUSION: These data suggest that PA-induced ICAM-1 expression and cell-cell adhesion in macrophages requires PKC-delta activation and that PKC-delta activation is triggers to sequential activation of p38 MAPK.
Blotting, Western ; Cell Adhesion ; Cell Line ; Endothelial Cells ; Flow Cytometry ; Humans ; Inflammation ; Intercellular Adhesion Molecule-1* ; Macrophages ; p38 Mitogen-Activated Protein Kinases ; Phosphatidic Acids ; Protein Kinase C ; Protein Kinases ; Second Messenger Systems ; Umbilical Veins

Although rare, renal arteriovenous fistulas have been discovered with increasing frequency since they were first described by Varela in 1923. Arteriovenous fistulas are classified as either congenital or acquired. The diagnosis of this condition is usually made by use of renal angiograms. In case of hematuria and/or hypertension either intra-arterial embolization or surgical excision and clipping of the arterial branch can be done. Intra-arterial superselective embolization seems to cause less functional loss of the renal parenchyme, whereas excision often leads to heminephrectomy or even total nephrectomy. Herein, we describe a case treated successfully in recurred renal A-V fistula after transcatheter embolization by Gianturco steel coil and reviewed the literature of renal A-V fistulas.
Arteriovenous Fistula ; Diagnosis ; Fistula* ; Hematuria ; Hypertension ; Kidney ; Nephrectomy ; Retreatment* ; Steel*

PURPOSE: The nitric oxide (NO) release by inducible nitric oxide synthase (iNOS) is the key events in macrophage response to lipopolysaccharide (LPS) which is suggested to be a crucial mediator for inflammatory and innate immune responses. NO is an important mediator involved in many host defense action and may also lead to a harmful host response to bacterial infection. However, given the importance of iNOS in a variety of pathophysiological conditions, control of its expression and signaling events in response to LPS has been the subject of considerable investigation. MATERIALS AND METHODS: The Raw264.7 macrophage cell line was used to observe LPS-stimulated iNOS expression. The expression of iNOS is observed by Western blot analysis and real-time RT-PCR. Protein kinase C (PKC)-alpha overexpressing Raw264.7 cells are established to determine the involvement of PKC-alpha in LPS-mediated iNOS expression. NF-kappaB activity is measured by IkappaBalpha degradation and NF-kappaB luciferase activity assay. RESULTS: We found that various PKC isozymes regulate LPS-induced iNOS expression at the transcriptional and translational levels. The involvement of PKC-alpha in LPS-mediated iNOS induction was further confirmed by increased iNOS expression in PKC-alpha overexpressing cells. NF-kappaB dependent transactivation by LPS was observed and PKC-alpha specific inhibitory peptide abolished this activation, indicating that NF-kappaB activation is dependent on PKC-alpha. CONCLUSION: Our data suggests that PKC-alpha is involved in LPS-mediated iNOS expression and that its downstream target is NF-kappaB. Although PKC-alpha is a crucial mediator in the iNOS regulation, other PKC isozymes may contribute LPS-stimulated iNOS expression. This finding is needed to be elucidated in further study.
Bacterial Infections ; Blotting, Western ; Cell Line ; I-kappa B Proteins ; Immunity, Innate ; Isoenzymes ; Luciferases ; Macrophages ; NF-kappa B ; Nitric Oxide ; Nitric Oxide Synthase Type II ; Protein Kinase C ; Protein Kinase C-alpha ; Protein Kinases ; Toll-Like Receptors ; Transcriptional Activation

Endomyocardial biopsy (EMB) is a valuable diagnostic procedure for the surveillance of cardiac allograft rejection. Interpretation of individual cases is still problematic due to variations of findings for grading of rejection and other associated lesions. We reevaluated an experience on endomyocardial biopsies to develop better diagnostic criteria for rejection and other complications. Immunohistochemical studies against cytokines were performed to assess the usefulness of the method for the diagnosis or researches. A total of 249 EMBs taken from 33 cardiac allograft recipients were reviewed. There were 25 males and 8 females. Dilated cardiomyopathy was present (24 cases) and valvular heart disease (4 cases), restrictive cardiomyopathy (3 cases) were also common conditions. We applied the grading system of the International Society for Heart Transplantation (ISHT) for the assessment of acute cellular rejection. Grades of 0, 1A, 1B, 2, 3A and 3B were 39.0%, 28.1%, 11.2%, 11.5%, 12.4% and 1.6% respectively, but 3.2% were inadequate. Thirty five episodes of grade 3A or 3B were present in 17 patients. The response to therapy was assessed using a next follow up biopsy, which revealed resolving or resolved rejection in 85% of patients. The intensity of immunohistochemical stains for IL-6 and TNF-alpha was increased in proportion to the histologic grade but Quilty lesion and cardiomyopathy also showed a positive reaction. The other pathologic findings were ischemic change, previous biopsy site, interstitial edema and fibrosis, and Quilty lesion. These findings showed usefulness of endomyocardial biopsy not only for the evaluation of cardiac allograft rejection but also for the diagnosis of associated cardiac lesions. Immunohistochemical study of the cytokines was related to the degree of inflammation rather than degree of rejection.
Allografts ; Biopsy* ; Cardiomyopathies ; Cardiomyopathy, Dilated ; Cardiomyopathy, Restrictive ; Coloring Agents ; Cytokines ; Diagnosis ; Edema ; Female ; Fibrosis ; Follow-Up Studies ; Heart Transplantation* ; Heart Valve Diseases ; Heart* ; Humans ; Inflammation ; Interleukin-6 ; Male ; Tumor Necrosis Factor-alpha

Spinal cord transection in children may occur following severe trauma such as a motor vehicle accident, and often without evidence of underlying skeletal injury. We report one case which showed cervical cord transection, where no evidence of underlying skeletal injury was seen on MR imaging, four weeks after trauma. When a neurologic deficit is present despite normal routine plain radiographs, further MR imaging is warranted to exclude a cord transection, as demonstrated in our patient.
Child ; Humans ; Magnetic Resonance Imaging* ; Motor Vehicles ; Neurologic Manifestations ; Spinal Cord Injuries

OBJECTIVE: To compare clinical results of laparoscopic assisted vaginal hysterectomy for uteri weighing 500 g or more with less than 500 g. And we compared clinical results between laparoscopic coagulation of uterine vessel (LH) and conventional LAVH. METHODS: We reviewed medical records of 296 patient who underwent LAVH from February 2004 to May 2006. They were divided into two groups, uteri weighing greater than 500 g and less than 500 g. And each group was divided into two groups, LH and conventional LAVH. Each groups were compared by operative time, hemoglobin change, complication, transfusion and hospital days. RESULTS: Operation time ,hemoglobin change on the 1st postoperative day and transfusion were significant greater in the uteri > or =500 g group than in the <500 g. However, there was no significant difference in hospital days, hemoglobin change on the 4th postoperative day and complication. In the <500 g group, LH group was lower than conventional LAVH group in operative time, hemoglobin change on the 1st and 4th postoperative day. In the > or =500 g group, there was no signicant difference in hospital days, operative time, hemoglobin change on the 1st postoperative day. However, hemoglobin change was smaller in the LH group than conventional LAVH group on the 4th postoperative day. CONCLUSION: This study demonstrates that despite the increased operating time and blood loss, LAVH can be safely performed for large uterus. However, surgeons need to be aware of high risk of blood transfusion. Modification of surgical method can decrease operating time and blood loss in LAVH.
Blood Transfusion ; Female ; Humans ; Hysterectomy ; Hysterectomy, Vaginal* ; Medical Records ; Operative Time ; Uterus*

Resveratrol is a polyphenolic compound in red wine that has anti-oxidant and cardioprotective effects in animal models. Reactive oxygen species (ROS) and monocyte chemotactic protein-1 (MCP-1) play key roles in foam cell formation and atherosclerosis. We studied LPS-mediated foam cell formation and the effect of resveratrol. Resveratrol pretreatment strongly suppressed LPS-induced foam cell formation. To determine if resveratrol affected the expression of genes that control ROS generation in macrophages, NADPH oxidase 1 (Nox1) was measured. Resveratrol treatment of macrophages inhibited LPS-induced Nox1 expression as well as ROS generation, and also suppressed LPS-induced MCP-1 mRNA and protein expression. We investigated the upstream targets of Nox1 and MCP-1 expression and found that Akt-forkhead transcription factors of the O class (FoxO3a) is an important signaling pathway that regulates both genes. These inhibitory effects of resveratrol on Nox1 expression and MCP-1 production may target to the Akt and FoxO3a signaling pathways.
Antioxidants/*pharmacology ; Cells, Cultured ; Chemokine CCL2/genetics/*metabolism ; Enzyme Activation/drug effects ; Foam Cells/*drug effects/physiology ; Forkhead Transcription Factors/metabolism ; Humans ; Lipopolysaccharides/pharmacology ; NADH, NADPH Oxidoreductases/genetics/*metabolism ; Proto-Oncogene Proteins c-akt/metabolism ; RNA, Messenger/metabolism ; Reactive Oxygen Species/*metabolism ; Signal Transduction ; Stilbenes/*pharmacology

The Jarcho-Levin syndrome (spondylothoracic dysplasia) is a rare autosomal recessive disorder characterized by a short neck, short trunk and a constricted thorax due to multiple rib and vertebral defects. The small size of thorax in newborns frequently leads to respiratory insufficiency and death in neonates or infancy. We reports the prenatal diagnosis using ultrasonography of a fetus affected with spondylothoracic dysplasia, or Jarcho-Levin syndrome, in patient without a positive family history for this condition.
Fetus ; Humans ; Infant, Newborn ; Neck ; Prenatal Diagnosis* ; Respiratory Insufficiency ; Ribs ; Thorax ; Ultrasonography*