Involvement of Protein Kinase C delta in Iron Chelator-Induced IL-8 Production in Human Intestinal Epithelial Cells / 대한해부학회지

Our previous study demonstrated that a bacterial siderophore, deferoxamine (DFO), could trigger inflammatory signals in human intestinal epithelial cells as a single stimulus, leading to IL-8 production via ERK1/2 and p38 phosphorylation and NF-kappa B-independent mechanism. In the present study, we proved that a novel protein kinase C (PKC)isoform, PKCdelta, is necessary for DFO-induced IL-8 production. Pretreatment of HT-29 cells with rottlerin showed remarkable inhibition of DFO-induced IL-8 production. In contrast, a conventional PKC inhibitor Go6976 did not show significant inhibition of DFO-induced IL-8 production. DFO induced strong phosphorylation of PKCdelta in the epithelial cells. Overexpression of PKCdelta resulted in enhanced PKCdelta phosphorylation, while transfection with dominant-negative PKCdelta vector failed DFO-induced phosphorylation. In addition, transfection of HT-29 cells with siRNA targeting endogenous PKCdelta, which suppressed PKCdelta expression, attenuated DFO-induced IL-8 production. These results demonstrate that PKCdelta plays an important role in regulating iron chelator-induced IL-8 production in human intestinal epithelial cells.

Ex Vivo Sentinel Node Mapping in Colorectal Cancer

PURPOSE: Lymph node analysis is essential for staging colorectal cancer. Intraoperative lymphatic mapping and sentinel lymphadenectomy remain to be investigated for most gastrointestinal neoplasms. Previous attempts to identify the sentinel node (SN) in solid tumors have used intraoperative techniques. This study describes a novel approach to identify the SN in colorectal cancer using ex vivo lymphatic mapping. METHODS: Eighty-two colorectal cancer patients underwent ex vivo lymphatic mapping and a sentinel lymph node biopsy using isosulfan blue dye following a standard surgical resection between March 2002 and September 2003. Within 5 minutes of resection, colorectal specimens were submucosally injected with isosulfan blue dye in four quadrants. Blue lymphatic channels were identified in the mesentery, and followed to the blue-stained SN(s), which were har vested. The specimens were fixed in formalin and subsequently analyzed in the usual fashion. In patients with T1 or T2 tumors, which were blue-stained nodes, but negative to hematoxylin and eosin staining, were further analyzed by serial section and immunohistochemical staining (IHC). RESULTS: At least one SN was identified in 79 patients of the 82 patients (96.3%). The average number of SNs identified per patient and nodes in each colorectal cancer specimen were 3 (range, 1~7) and 17.1 (range: 11~47). Thirty five patients had lymph nodes containing a metastatic disease. Thirteen patients had metastases in both sentinel and nonsentinel nodes. There were 7 sentinel lymph nodes as the only site of metastatic disease. In 15 patients the sentinel nodes were negative for disease, whereas the nonsentinel lymph nodes contained a metastatic disease (false negative rate = 42.9%). The false negative rates of SN(s) metastasis in the 26 patients with T1 or T2 tumors were 16.7 and 7.7% by H&E and by serial section and IHC. CONCLUSION: Ex vivo mapping of the colon is technically feasible, and may provide a useful approach to evaluate lymph node metastasis in patient with T1 or T2 colorectal cancers.

Effects of Sulphasalazine and Glucocorticoid on the Regulation of CCL20 Gene Expression in the Peripheral Blood Cells of Korean Patients with Ulcerative Colitis / 대한해부학회지

Discovery of Nod2 has brought to light the significance of mononuclear cells as well as epithelial cells in inflammatory bowel disease (IBD) pathogenesis. Similarly, CCL20 is expressed in both mononuclear cells and epithelial cells and is likely to link innate and acquired immunity. We therefore asked whether CCL20 expression is altered in the peripheral blood mononuclear cells (PBMCs) from patients with ulcerative colitis (UC), a major type of IBD in Korea, and is correlated with the disease activity. The expression levels of CCL20 mRNA were significantly high in the PBMCs from the patients with UC. CCL20 protein expression was also up-regulated in the mucosal epithelium in UC but not in normal controls. Interestingly, however, disease activity index (DAI) revealed that untreated UC groups express higher expression levels of CCL20 mRNA than treated UC groups, implying that CCL20 may be a potential target for the anti-inflammatory treatments. In an agreement with this, three months follow up study revealed that the UC patients who were treated with 5-amino salicylic acid (5-ASA) and glucocorticoid showed dramatic decrease in their CCL20 mRNA levels as compared to untreated ones. Moreover, TNF-alpha-or IL-1beta-induced CCL20 secretion in human epithelial HT-29 cells was significantly diminished by the treatment with 5-ASA and/or dexamethasone, suggesting that CCL20 may be one of the central targets of the anti-inflammatory drugs. Collectively, these results suggest that CCL20 expression in UC may be associated with altered immune and inflammatory responses in the blood as well as the intestinal mucosa and further implied a potential for CCL20 as an important diagnostic marker for UC.

Iron Chelator Induces MIP-3alpha/CCL20 in Human Intestinal Epithelial Cells: Implication for Triggering Mucosal Adaptive Immunity / 체질인류학회지

A previous report by this laboratory demonstrated that bacterial iron chelator (siderophore) triggers inflammatory signals, including the production of CXC chemokie IL-8, in human intestinal epithelial cells(IECs). Microarray-based gene expression profiling revealed that iron chelator also induces CC chemokie MIP-3alpha/CCL20. As CCL20 is chemotactic for the cells involved in host adaptive immunity, this suggests that iron chelator may stimulate IECs to have the capacity to link mucosal innate and adaptive immunity. The basal medium from iron chelator deferoxamine (DFO)-treated HT-29 monolayers was as chemotactic as rhCCL20 at equivalent concentrations to attract CCR6+cells. The increase of CCL20 protein secretion appeared to correspond to that of CCL20 mRNA levels, as determined by real-time quantitative RT-PCR. The efficacy of DFO at inducing CCL20 mRNA was also observed in human Peripheral Blood Mononuclear Cells (PBMCs) and in THP-1 cells, but not in Human Umbillical Vein Endothelial Cells (HUVECs). Interestingly, unlike other proinflammatory cytokines, such as TNF-alpha and IL-1beta, a time-dependent experiment revealed that DFO slowly induces CCL20, suggesting a novel mechanism of action. A pharmacologic study also revealed that multiple signaling pathways are differentially involved in CCL20 production by DFO, while some of those pathways are not involved in TNF-alpha-induced CCL20 production. Collectively, these results demonstrate that, in addition to some bacterial products known to induce host adaptive immune responses, direct chelation of host iron by infected bacteria may also contribute to the initiation of host adaptive immunity in the intestinal mucosa.

Pathologic Findings in the Liver of Hepatitis B Virus X Transgenic Mice

BACKGROUND: The aim of this study was to investigate the hepatic pathology of HBx transgenic mice. METHODS: The gross and histological examinations were done in 125 HBx transgenic mice and 34 non-transgenic littermates. RESULTS: The incidence of a hepatic tumor was in-creased in the HBx transgenic mice older than 7 months and the overall incidence of a hepatic tumor was 62.2% (51/82) in the 13-18 months group of the HBx transgenic mice. The size of the hepatic tumor was 2.06+/-.92 mm in the 7-12 months group and 4.94+/-.05 mm in the 13-18 months group of HBx transgenic mice. All hepatic tumors were hepatocellular carcinomas and the histological patterns of hepatocellular carcinoma were either solid (84.2%, 48/57) or trabecular (15.8%, 9/57). Dysplastic changes in the hepatocytes were evident in 59.2% (74/125) of the HBx transgenic mice. There was lymphocyte infiltration, necrosis, fatty metamorphosis in both the dysplastic and tumor areas of the HBx transgenic mice. Vascular ectasia was identified in the tumor area of the HBx transgenic mice. CONCLUSIONS: The pathological findings of the HBx transgenic mice were dysplastic changes in the hepatocytes and development of a hepatocellular carcinoma associated with lymphocyte infiltration, necrosis, fatty metamorphosis in the dysplastic area and tumor area of the HBx transgenic mice.

Fine Needle Aspiration Cytology of Proliferative Fasciitis: A Case Report / 대한세포병리학회지

Fine needle aspiration cytology (FNAC) is an easy convenient non-invasive method in the diagnosis of superficial palpable masses. The cytologic findings by FNAC of reactive and neoplastic lesions in various organs including breast, lymph node, thyroid, salivary gland, etc., have been described, but, those of soft tissue lesions including proliferative fasciitis are relatively rare to find. We recently experienced a case of FNAC of proliferative fasciitis in the left back of a 72-year-old male. The FNAC smears were scant in cellularity and contained large cells with abundant basophilic cytoplasm, one to two nuclei lying at the periphery, and prominent nucleoli that resemble ganglion cells.

A Study on the Expression of Proliferating Cell Nuclear Antigen and Apoptosis of the Hepatocellular Carcinoma in Human and Hepatitis B Virus X Transgenic Mice

BACKGROUND: This experiment was designed to study the cell kinetics of hepatocellular carcinoma (HCC) in both hepatitis B virus X (HBx) transgenic mice and humans. METHODS: The immunohistochemical stain of proliferating cell nuclear antigen (PCNA) and TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay of apoptosis were used on formalin fixed-paraffin embedded tissues. RESULTS: PCNA labeling indices (PCNA-LI) in the liver of HBx transgenic mice were markedly increased in HCC (11.3%) compare to the dysplastic areas (1.3%) and in the liver of non-transgenic littermates (0.1%). There was no significant difference of PCNA-LI in the dysplastic areas between HCC developed mice and non-HCC developed mice. Apoptosis labeling indices (Apoptosis-LI) in both the dysplastic areas and HCC of HBx transgenic mice were similar to those of non-transgenic littermates. PCNA-LI was markedly increased in human HCC (28.9%) compare to the background of HCC (2.9%) and the control liver (2.9%). Apoptosis-LI was decreased in human HCC (0.3%) compare to the background of HCC (0.4%) and the control liver (1.0%). Conclusion : There is a marked increase of cell proliferating activity in human HCC and in HCC of HBx transgenic mice, and there is a decrease of apoptosis in human HCC, but not in HCC of HBx transgenic mice.

Primary Transitional Cell carcinoma of the Ovary / 대한부인종양콜포스코피학회잡지

Primary transitional cell carcinoma of the ovary is recently recognized, as one of the main types of ovarian carcinoma. Histologically, it is distinguished from malignant Brenner tumor only in the abscence of benign or proliferative Brenner tumor component. primary transitional cell carcinomas are more aggressive than malignant Brenner tumors. However, Primary transitional cell carcinomas have a better response to chemotherapy than other types of ovarian carcinomas. We report a case of primary transitional cell carcinoma presenting as both ovarian masses that developed in a 60-year-old woman with a brief review of literature.