Ethyl acetate fraction from Cudrania tricuspidata inhibits IL-1beta-stimulated osteoclast differentiation through downregulation of MAPKs, c-Fos and NFATc1.

BACKGROUND/AIMS: The present study was performed to determine the effects of the ethyl acetate extract of Cudrania tricuspidata (EACT) on interleukin (IL)-1beta-stimulated receptor activator of NF-kappaB ligand (RANKL)-mediated osteoclast differentiation. METHODS: Bone marrow cells were harvested from 6-week-old male imprinting control region mice, and the differentiation of osteoclasts from these cells was evaluated by tartrate-resistant acid phosphatase and resorption pit formation assay. Phosphorylated extracellular signal regulated kinase (p-ERK), phosphorylated p38, phosphorylated c-Jun amino-terminal kinase, NF-kappaB (p65), IkappaBalpha, c-Fos, and nuclear factor of activated T-cells c1 (NFATc1) expression was examined by immunoblotting and quantitative reverse transcription-polymerase chain reaction. RESULTS: EACT inhibits IL-1beta-stimulated RANKL-mediated osteoclast differentiation. EACT also inhibits IL-1beta-stimulated RANKL-mediated phosphorylation of ERK 1/2, p38 mitogen activated protein kinase, and expression of c-Fos and NFATc1. CONCLUSIONS: These results suggest that EACT may be involved in the inhibition of bone loss by preventing osteoclast formation and may be used to manage bone destruction in inflammatory diseases, such as rheumatoid arthritis.

Adrenomedullin inhibits MAPK pathway-dependent rheumatoid synovial fibroblast-mediated osteoclastogenesis by IL-1 and TNF-alpha.

The objective of this study is to determine the effects of adrenomedullin (AM) on IL-1- and TNF-alpha-induced rheumatoid synovial fibroblasts (RASFs)-mediated osteoclastogenesis. The formation of osteoclasts in co-cultures of RASFs and peripheral blood mononuclear cells was evaluated by tartrate-resistant acid phosphatase and resorption pit formation assay. The expression of RANKL, OPG, p-ERK, p-p38, and p-JNK was examined by immunoblotting and quantitative reverse transcription-polymerase chain reaction. AM (1-52) inhibits IL-1- and TNF-alpha-induced RASFs-mediated osteoclastogenesis. AM affected IL-1-, TNF-alpha-induced RANKL and OPG expression in RASFs. AM also inhibits IL-1 and TNF-alpha-induced phosphorylation of ERK-1/2, p38 MAPK, and JNK. Inhibitor of AM (AM 22-52) inhibits the effects of AM on the osteoclastogenesis. These results suggest that AM might be involved in the inflammatory cytokines-mediated osteoclastogenesis and thus bone damage, and indicate that it can be a new therapeutic strategy against joint destruction in RA.

Molecular scintigraphic imaging using 99mTc-transferrin is useful for early detection of synovial inflammation of collagen-induced arthritis mouse.

Transferrin receptor (TfR) is highly expressed on rapidly dividing inflammatory cells, but not in nonproliferating cells. We investigated whether scintigraphic imaging using 99mTechnetium-radiolabeled transferrin (99mTc-Tf) is useful for early detection of synovial inflammation of collagen-induced arthritis (CIA) mouse. 99mTc-Tf conjugate was synthesized to target the TfR in inflamed synovium. 99mTc-Tf scintigraphic images were obtained in nonarthritic normal mouse and advanced phase of CIA mouse. 99mTc-Tf and 99mTc-methylenediphosphonate (99mTc-MDP) bone scintigraphic images were obtained in same early phase of CIA mouse. Western blot analysis, hematoxylin & eosin (H&E), and immunohistochemical staining were performed to determine development of arthritis and expression of TfR. Image analyses revealed that the uptake of 99mTc-Tf in inflamed joints of advanced phase of CIA mouse was markedly higher than those by normal nonarthritic mouse. 99mTc-Tf scintigraphy also showed higher uptake in knee joint prior to significant joint swelling in early phase of CIA mouse but 99mTc-MDP bone scintigraphy does not. These scintigraphic findings are well correlated with the results of Western blot, H&E and immunohistochemical analysis. In conclusion, TfR can be used as a specific target for molecular imaging in CIA mouse, and 99mTc-Tf scintigraphy detects synovial inflammation prior to significant clinical findings in CIA mouse.

Epigallocatechin-3-gallate suppresses TNF-alpha -induced production of MMP-1 and -3 in rheumatoid arthritis synovial fibroblasts.

Rheumatoid arthritis (RA) synovial fibroblasts produce matrix metaloproteinases (MMPs), which destroy cartilage and bone in RA joint. Tumor necrosis factor-alpha (TNF-alpha) is one of the most important mediator leading to MMP production in RA synovial fibroblasts. Here we show that epigallocatechin-3-Gallate (EGCG) suppresses TNF-alpha-induced production of MMP-1 and MMP-3 in RA synovial fibroblasts, which was accompanied by inhibition of mitogen activated protein kinase (MAPK) and activator protein-1 (AP-1) pathways. EGCG treatment resulted in dose-dependent inhibition of TNF-alpha-induced production of MMP-1 and MMP-3 at the protein and mRNA levels in RA synovial fibroblast. EGCG treatment also inhibited TNF-alpha-induced phosphorylation of MAPKs, such as ERK1/2, p38, JNK. Electrophoretic mobility shift assay revealed that EGCG inhibits binding of AP-1 proteins to its response elements in synovial fibroblast treated. Thus, EGCG may play a role in regulating inflammation and bone destruction in RA patients.

Absence of antibody to cyclic citrullinated peptide in sera of non-arthritic patients with chronic hepatitis B virus infection.

The objective of this study was to investigate if antibody to cyclic citrullinated peptide (anti-CCP) is detected in sera of patients with chronic hepatitis B virus (HBV) infection. Serum anti-CCP and IgA, IgG, and IgM rheumatoid factor (RF) isotypes were measured by enzyme-linked immunosorbent assay on 176 non-arthritic patients with HBV infection. IgA RF, IgG RF, and IgM RF were detectable in 29.5, 21, and 18.8% of the tested sera, respectively, with a total seropositivity rate of 42.7%. Marginally elevated anti-CCP was detected in one patient (0.6%). By regression analysis, there was no statistically significant association between the serum levels of anti-CCP and serum IgA, IgG, or IgM RF (R (2) = 0.033, with respective p values of 0.224, 0.297, and 0.334). In conclusion, anti-CCP was rarely detected in non-arthritic patients with HBV infection in contrast to RF. Thus, testing for anti-CCP may be a useful tool for the diagnosis of rheumatoid arthritis in this population.

CD40 ligation of rheumatoid synovial fibroblasts regulates RANKL-mediated osteoclastogenesis: evidence of NF-kappaB-dependent, CD40-mediated bone destruction in rheumatoid arthritis.

OBJECTIVE: To determine whether CD40 ligation of rheumatoid arthritis synovial fibroblasts (RASFs) is able to induce RANKL expression and osteoclastogenesis in RASFs, and to identify its mechanism of action in patients with RA. METHODS: CD40 of RASFs was ligated with CD40 ligand (CD40L)-transfected L cells or activated T cells. The formation of osteoclasts in cocultures of CD40-ligated RASFs and T lymphocyte-depleted peripheral blood mononuclear cells was evaluated by tartrate-resistant acid phosphatase staining, detection of calcitonin receptor, and resorption pit formation assay. The expression of NF-kappaB, IkappaB alpha, ERK-1/2, phospho-ERK-1/2, p38, phospho-p38, and RANKL was examined by immunoblotting and/or semiquantitative reverse transcription-polymerase chain reaction. RESULTS: CD40 ligation of RASFs by CD40L-transfected L cells or activated T cells induced RANKL expression and enhanced osteoclastogenesis. CD40 ligation of RASFs also induced activation of ERK-1/2, p38 MAPK, and NF-kappaB and up-regulation of CD40 ligation-induced RANKL expression, whereas osteoclastogenesis was reduced in RASFs transfected with a dominant-negative mutant of IkappaB alpha or by an NF-kappaB inhibitor. However, specific inhibitors of MAPK/ERK-1/2 and p38 MAPK partially blocked the induction of RANKL expression and osteoclastogenesis. Monoclonal antibodies against interleukin-1 and tumor necrosis factor alpha partially inhibited CD40 ligation-mediated osteoclastogenesis. CONCLUSION: These results indicate that CD40 ligation of RASFs induces RANKL expression mainly via NF-kappaB activation and also results in enhanced osteoclast formation, both of which might play important roles in bone and cartilage destruction in RA. Inhibition of the CD40-CD40L interaction is a potential strategy for the prevention of bone damage in RA.

Cytologic Findings of Parathyroid Carcinoma: Report of Two Cases / 대한세포병리학회지

Parathyroid carcinoma is a rare disorder accounting for 0.5% to 5% of parathyroid neoplasia. Diagnosis of parathyroid carcinoma in fine needle aspiration cytology(FNAC) is difficult because all characteristic features of parathyroid carcinoma can be recognized in parathyroid adenoma or hyperplasia. Cellular atypism cannot be used for the diagnostic criteria of parathyroid carcinoma as malignancies of most other organs. We experienced two cases of cytologic features of parathyroid carcinoma confirmed by histologic examination. The majority of tumor cells formed large cohesive clusters, although individual tumor cells were also present. The tumor cells displayed rather pleomorphic round to oval nuclei, occasional prominent nucleoli, and distinct cytoplasmic margin. Occasionally karyolysis, anuclear cells, and nonepithelial cell clusters were noted. The histologic findings showed a partially lobulated architecture, with admixture of sheets of chief cells, oxyphil cells, and occasional water clear cells. The tumor infiltrated into the thyroid parenchyme and perithyroidal soft tissue. The electron microscopic study of case 1 disclosed typical findings of parathyroid neoplasm; clusters of secretory chief cells with centrally located round to ovoid nuclei, moderately clumped heterochromatins and one or two nucleoli. The tumor cells showed conspicous interdigitation of contiguous cell membrane and intercellular microvilli.

Expression of Cyclin B1 and cdc2 in Nodal Non-Hodgkin's Lymphoma and its Prognostic Implications

To investigate the role of cyclin B1 and cdc2 in the pathogenesis and progression of malignant lymphoma, 68 cases of nodal non-Hodgkin's lymphoma were examined about the expression of cyclin B1 and cdc2 along with p53 and Ki-67 by immunohistochemical method. The correlation of their expression with various clinicopathologic findings was also analyzed. Cyclin B1 and cdc2 were diffusely expressed in 39 cases (57.4%) and 54 cases (79.4%) out of 68 cases studied, respectively. The mean labeling indices of cyclin B1 and cdc2 in malignant lymphoma were 31.9% and 68.0%, respectively. In normal lymphoid tissues, cyclin B1 and cdc2 were expressed predominantly in the germinal center with mean labeling indices of 13.9% and 28.3%, respectively. The correlation between the expression of cyclin B1 and cdc2 was noted (p=0.013). The expression of Ki-67 was correlated with that of cyclin B1 (p=0.023) and marginally correlated with that of cdc2 (p=0.056). The expression of cdc2 and p53 in complete remission group to chemotherapy was lower than that of progressive disease group (p=0.047, p=0.049). In multivariate analysis, the clinical stage alone showed significance on overall survival (p=0.049). In conclusion, cyclin B1 and cdc2 appeared to be involved in the genesis or progression of malignant lymphoma and cdc2 can be a useful marker for response to chemotherapy.

Cytologic diagnosis of a chordoma without physaliferous cells: A case report / 대한세포병리학회지

Cytologic features of conventional chordoma have been described and most reports emphasize the presence of large cells with numerous well defined cytoplasmic vacuoles or physaliferous cells. We report fine needle aspiration cytologic (FNAC) findings of a case of chordoma without physaliferous cells. The smear was cellular and composed of large cohesive clusters or individually scattered cells in mucinous background. The round or cuboidal cells had centrally located nuclei with fine granular chromatin, inconspicuous nucleoli, and occasional vacuolated cytoplasm. Mild to moderate pleomorphism was noted. Physaliferous cells are extremely helpful when present in cytologic material, but they are not necessary for diagnosis. Thus clinical history, roentgenographic appearance, and exact location of the lesion are required for the successful interpretation of presacral aspirates together with cytologic findings.

Histopathologic Analysis of Malignant Lymphoma Involving the Skin and Its Relationship with the Epstein-Barr Virus

The author classified 38 cases of malignant lymphoma involving the skin primarily or secondarily by the new WHO classification with minor modifications and carried out RNA in situ hybridization and/or polymerase chain reaction (PCR) to investigate the role of Epstein-Barr virus (EBV). A case was follicular lymphoma of B cell origin and 37 cases were malignant lymphomas of T cell origin, including 15 cases of Mycosis fungoides/Sezary syndrome, five cases of subcutaneous panniculitis-like T cell lymphomas, a case of anaplastic large cell lymphoma, and four cases of primary cutaneous CD30 T cell lymphoproliferative disorders. There were eight cases of unspecified peripheral T cell lymphomas, in which four cases were composed of medium-sized cells, three cases of large cells, and a case of lymphoepithelioid cells. Four cases of nasal and nasal type NK/T cell lymphomas and three cases of unspecified peripheral T cell lymphomas showed EBV genome. The nasal and nasal type NK/T cell lymphomas, especially those involving the nasal cavity, showed close association with the EBV infection.

Diagnostic criteria for malignancy in bile cytology and its usefulness

Fifty three bile specimens from 42 patients were reviewed to assess the diagnostic role of the bile cytology and to define more reliable cytologic indicators of malignancy. Forty three bile specimens came from 34 patients with malignant biliary strictures and 10 bile specimens were from eight patients with benign conditions. There were no false positives. The diagnostic specificity of bile cytology was 100% while diagnostic sensitivity was 55.8%. Overall diagnostic accuracy was 64.2%. We identified four key criteria as cytologic indicators of malignancy among 20 variables by using multiple regression analysis: loss of honeycomb arrangement, hyperchromatism, increased N/C ratio, and coarse chromatin. When bile specimens with three or more of these four criteria are thought to represent malignancy, the sensitivity of diagnosis of malignancy was 65.2%, specificity was 90% and diagnostic accuracy was 69.8%.