• Clinical and Experimental Pediatrics
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• v.53 no.4
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• pp.525-531
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• 2010

Purpose : Expression levels of tumor necrosis factor (TNF)-${\alpha}$ expression on the mucosa of the small intestine is increased in patients with villous atrophy in food protein-induced enterocolitis syndrome (FPIES). TNF-${\alpha}$ has been reported to induce apoptotic cell death in the epithelial cells. We studied the TNF family and TNF-receptor family apoptosis on the duodenal mucosa to investigate their roles in the pathogenesis of FPIES. Methods : Fifteen infants diagnosed as having FPIES using standard oral challenge test and 5 controls were included. Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining was performed to identify the apoptotic cell death bodies. Immunohistochemical staining of TNF-${\alpha}$, Fas ligand (FasL) for TNF family and TNF-related apoptosis-including ligand (TRAIL) receptor 1 (DR4), TRAIL receptor 2 (DR5), and Fas for TNF-receptor family were performed to determine the apoptotic mechanisms. Results : $TUNEL^+$ was significantly more highly expressed in the duodenal mucosa of FPIES patients than in controls ($P$-0.043). TNF-${\alpha}$ ($P$=0.0001) and DR4 ($P$=0.003) were significantly more highly expressed in FPIES patients than in controls. Expression levels of FasL, Fas, and DR5 were low in both groups and were not significantly different between the 2 groups. Conclusion : These results suggest that FPIES pathogenesis is induced by apoptosis, and that TNF-${\alpha}$ expression and DR4 pathway may have an important role in apoptosis.

• The Journal of Korean Obstetrics and Gynecology
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• v.23 no.3
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• pp.56-77
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• 2010

Purpose: This study was performed to investigate effects of Boyanghwano-Tang on the development of experimentally-induced endometriosis in rats. Methods: Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and divided them into three groups: (1) sham-operated group(n=8), (2) surgically induced endometriosis and untreated control group (n=8), (3) surgically induced endometriosis and Boyanghwano-Tang treated group. Boyanghwano-Tang was orally administrated for 15 days after operation. Then we measured the body weight, the volume of endometriotic implants, the weight of uterus and ovary, and investigated the concentration of cytokines (MCP-1, TNF-$\alpha$, IL-$1{\beta}$, IL-6) in peritoneal fluids. Histopathology, immunohistochemistry for COX-2 and VEGF, and histochemistry for mast cell in transplanted uterine tissue were performed. Results: - The volume($mm^3$) of endometriotic implants in Boyanghwano-Tang treated group ($122.3{\pm}45.0$) was significantly decreased(p<0.05) compared with control group($222.1{\pm}109.1$). - The concentration(pg/$m{\ell}$) of MCP-1 in peritoneal fluids in Boyanghwano-Tang treated group($1026.3{\pm}196.5$) was significantly decreased(p<0.05) compared with control group($1412.5{\pm}345.7$). - The concentration(pg/$m{\ell}$) of TNF-$\alpha$ in peritoneal fluids in Boyanghwano-Tang treated group($936.5{\pm}94.3$) was significantly decreased(p<0.01) compared with control group($1126.2{\pm}139.9$). - The concentration(pg/$m{\ell}$) of IL-$1{\beta}$ in peritoneal fluids in Boyanghwano-Tang treated group($78.5{\pm}13.3$) was significantly decreased(p<0.01) compared with control group($105.3{\pm}17.6$). - The concentration(pg/$m{\ell}$) of IL-6 in peritoneal fluids in Boyanghwano-Tang treated group($107.4{\pm}15.8$) was decreased compared with control group($122.8{\pm}19.3$). - Histopathologically, proliferation of endometriotic epithelia, infiltration of inflammatory cells and angiogenesis in transplanted uterine tissue of Boyanghwano-Tang treated group were weakly observed than those of control group. - The percentage of positive epithelial layers for COX-2 in Boyanghwano-Tang treated group($51.5{\pm}14.1$) was significantly decreased(p<0.01) compared with control group($75.1{\pm}16.3$). - The VEGF expression of endometriotic epithelia, neovascular endothelia and stromal cells in transplanted uterine tissue of Boyanghwano-Tang treated group were weakly observed than those of control group. - The number of mast cells in adjacent tissue of transplanted uterine tissue in Boyanghwano-Tang treated group($75.9{\pm}13.1$) was decreased compared with control group($91.0{\pm}28.3$). Conclusion: On the basis of these results, we concluded that Boyanghwano-Tang have inhibiting effects on the development of transplanted uterine tissue. And these effects may be related with decreased production of MCP-1, TNF-$\alpha$, IL-$1{\beta}$ and decreased expression of COX-2 and VEGF by administration of Boyanghwano-Tang.

• Restorative Dentistry and Endodontics
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• v.31 no.6
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• pp.437-444
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• 2006

This study was aimed to elucidate the biological function of OD314 (Apin protein), which is related to ameloblast differentiation and amelogenesis. Apin protein, calcifying epithelial odontogenic (pindborg) tumors (CEOTs)-associated amyloid, were isolated from CEOTs, and has similar nucleotide sequences to OD314. We examined expression of the OD314 mRNA using in-situ hybridization during tooth development in mice. Expression of OD314 and several enamel matrix proteins were examined in the cultured ameloblast cell line up to 28 days by reverse transcription-polymerase chain reaction (RT-PCR) amplification. After inactivation and over-expression of the OD314 gene in ameloblast cell lines using U6 vectordriven RNA interference and CMV-OD314 construct, RT-PCR were performed to evaluate the effect of the OD314 during amelogenesis. The results were as follows: 1. In in-situ hybridization, OD314 mRNAs were more strongly expressed in ameloblast than odontoblast. 2. When ameloblast cells were cultured in the diffcrentiation and mineralization medium for 28 days, the tuftelin mRNA expression was maintained from the beginning to day 14, and then gradually decreased to day 28. The expressions of amelogenin and enamelin were gradually decreased according to the ameloblast differentiation. 3. Inactivation of OD314 by U6-OD314 siRNA construct down-regulated the expression of OD314, MMP-20, and tuftelin, whereas over-expression of OD314 by CMV-OD314 construct up-regulated the expression of OD314 and MMP-20 without change in tuftelin. These results suggest that OD314 is considered as an ameloblast-enriched gene and may play the important roles in ameloblast differentiation and mineralization.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.3
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• pp.207-216
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• 2010

Objective: This study was performed to clarify the role of HomeoboxA (HOXA) and its related signaling molecules in the decidualization of primary cultured endometrial cells. Methods: Human endometrial tissues were obtained by curettage of hysterectomy specimens from patients with conditions other than endometrial diseases. Tissues were minced and digested with Trypsin-EDTA for 20 min, $37^{\circ}C$. Cells were cultured with DMEM/F12 medium in $37^{\circ}C$, 5% $CO_2$ incubator for 24 hrs. Cells were treated with HOXA10 siRNA and added transforming growth factor (TGF)-${\beta}1$ (10 ng/mL) for 48 hrs to induces decidualization in vitro. Reverse transcription polymerase chain reaction analysis was accomplished to observe the expression of HOXA10, prolactin, cyclooxygenase (COX)-2, peroxisome proliferatoractivated receptor (PPAR)-$\gamma$, and wingless-type MMTV integration site family (Wnt). Results: HOXA10 expression was increased (1.8 fold vs. non-treated control) in TGF-${\beta}1$ treated cells. Decidualization marker, prolactin, was significantly increased in TGF-${\beta}1$ treated cells compared with HOXA10 siRNA treated cells. Endometrial cell differentiation marker, COX-2 was down-regulated by HOXA10 siRNA even if cells were treated with TGF-${\beta}1$. Wnt4 was down-regulated by treated with HOXA10 siRNA, this expression patters was not changed by TGF-${\beta}1$. Expression of PPAR-$\gamma$ was down regulated by TGF-${\beta}1$ in regardless of HOXA10 siRNA treatment. Conclusion: TGF-${\beta}1$ which is induced by progesterone in endometrial epithelial cells may induces stromal cell decidualization via HOXA10 and Wnt signaling cascade.

• The Journal of Korean Obstetrics and Gynecology
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• v.21 no.3
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• pp.34-59
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• 2008

Purpose: This study was performed to investigate the inhibiting effects of Gungguijohyeoleum on the development of experimentally-induced endometriosis in rats. Methods: Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and we divided them into three groups: (1) sham-operated group(n=8), (2) surgically induced endometriosis and untreated control group(n=8), (3) surgically induced endometriosis and Gungguijohyeoleum treated group(n=8), Gungguijohyeoleum was orally administrated for 15 days after operation. Then we measured the body weight, the volumes of endometriotic implants, the weight of uterus and ovary, and investigated the concentrations of cytokines(MCP-1, $TNF-{\alpha}$, $IL-1{\beta}$, IL-6) in peritoneal fluids. Histopathology, immunohistochemistry for COX-2 and VEGF, and histochemistry for mast cell in transplanted uterine tissue were performed. Results: - The volume ($mm^3$) of endometriotic implants in Gungguijohyeoleum treated group ($76.4{\pm}24.5$) was significantly decreased(p<0.05) compared with control group($222.1{\pm}109.1$). - The concentration (pg/ml) of MCP-1 in peritoneal fluids in Gungguijohyeoleum treated group ($1060.8{\pm}280.8$) was significantly decreased(p<0.05) compared with control group($1412.5{\pm}345.7$). - The concentration (pg/ml) of $TNF-{\alpha}$ in peritoneal fluids in Gungguijohyeoleum treated group ($827.5{\pm}219.9$) was significantly decreased(p<0.01) compared with control group($1126.2{\pm}139.9$). - The concentration (pg/ml) of $IL-1{\beta}$ in peritoneal fluids in Gungguijohyeoleum treated group($83.4{\pm}9.0$) was significantly decreased(p<0.01) compared with control group($105.3{\pm}17.6$). - The concentration (pg/ml) of IL-6 in peritoneal fluids in Gungguijohyeoleum treated group($108.9{\pm}15.7$) was decreased compared with control group($122.8{\pm}19.3$). - Histopathologically, proliferation of endometriotic epithelia, infiltration of inflammatory cells and angiogenesis in transplanted uterine tissue of Gungguijohyeoleum treated group were weakly observed than those of control group. - The percentage(%) of positive epithelial layers for COX-2 in Gungguijohyeoleum treated group($56.8{\pm}12.8$) was significantly decreased(p<0.01) compared with control group($75.1{\pm}16.3$). -The VEGF expression of endometriotic epithelia, neovascular endothelia and stromal cells in transplanted uterine tissue of Gungguijohyeoleum treated group was weakly observed than that of control group. - The numbers of mast cells in transplanted uterine tissue in Gungguijohyeoleum treated group($35.4{\pm}18.4$) were significantly decreased(p<0.01) compared with control group ($91.0{\pm}28.3$). Conclusion: On the basis of these results, we concluded that Gungguijohyeoleum has inhibiting effects on the development of transplanted uterine tissue. And these effects may be related with decreased production of MCP-1, $TNF-{\alpha}$, $IL-1{\beta}$, COX-2, VEGF and mast cells by administration of Gungguijohyeoleum.

• Journal of Sericultural and Entomological Science
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• v.25 no.2
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• pp.1-31
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• 1984

Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.

• Tuberculosis and Respiratory Diseases
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• v.45 no.2
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• pp.290-300
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• 1998

Background: CYFRA 21-1 is a tumor marker which measures a fragment of cytokeratin 19 expressed by epithelial cells in bronchus. It is known that cytokeratin 19 is abundant in squamous epithelial cell cancer of the lung. However, if the incidence of elevated serum CYFRA 21-1 level in patients with benign lung diseases or pulmonary tuberculosis with severe parenchymal damage is high the specificity of CYFRA 21-1 could be decreased. The purpose of this study is to investigate the changes of serum CYFRA 21-1 according to the degree of parenchymal damage and the usefulness of CYFRA 21-1 for diagnosing possibly combined lung cancer in patients with pulmonary tuberculosis. Method: We studied the changes of serum CYFRA 21-1 according to the sputum AFB stain, radiologic manifestation and history of treatment in 81 patients with pulmonary tuberculosis, and 20 healthy persons, 25 patients with lung cancer, as a control group. CYFRA 21-1 concentration in serum was quantified by the immunoradiometry assay(Centocor$^{(R)}$). Result: The results were as follow; Serum CYFRA 21-1 level was significantly lower in patients with pulmonary tuberculosis($1.54{\pm}1.19ng/mL$, p<0.01) as compared to patients with lung cancer($12.25{\pm}15.97ng/mL$), and was slightly higher than the level in heathy persons($0.90{\pm}0.49ng/mL$) but there was no significant difference. Serum CYFRA 21-1 level was below the cut-off value of 3.3ng/mL in 95 percent of patients with pulmonary tuberculosis but it was above the cut-off value in 64 percent of patients with lung cancer. Serum CYFRA 21-1 level was significantly higher in the initial treatment group($1.91{\pm}1.55ng/mL$, p<0.05) as compared to the treatment. failure group ($0.92{\pm}0.30ng/mL$). According to the sputum AFB smear, serum CYFRA 21-1 level in patients with negative result was slightly higher than the level in patients with positive result but there was no significant difference. According to the radiologic manifestation, serum CYFRA 21-1 level was significantly higher in patients with infiltrative lesion ($2.15{\pm}1.63ng/mL$, p<0.01) as compared to patients with destructive lesion ($l.04{\pm}0.54ng/mL$). As the size of cavity or destructive lesion was larger, the level was significantly lower(p<0.05). Conclusion: As serum CYFRA 21-1 level was significantly higher in the initial treatment group and patients with infiltrative lesion, it suppose to be closely related with the degree of parenchymal damage of the lung of the pulmonary tuberculosis. However CYFRA 21-1 could be useful method for diagnosing lung cancer even in patients with pulmonary tuberculosis combined with lung cancer because of the fact that it was below the cutoff value of 3.3ng/mL in 95 percent of patients with pulmonary tuberculosis.

• Applied Microscopy
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• v.23 no.2
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• pp.27-40
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• 1993

This study was taken to examine the light microscopic and ultrastructural changes of gill and kidney of female tilapia{Oreochromis niloticus) adapted in 0%o, 10%o, 20%o, and 30%o salt concentrations, respectively, by light, scanning and transmission electron microscope. The results obtained in these experiments were summarized as follows: Gill chloride cell hyperplasia, gill lamellar epithelial separation, kidney glomerular shrinkage, blood congestion in kidneys and deposition of hyalin droplets in kidney glomeruli, tubules were the histological alterations in Oreochromis niloticus. Incidence and severity of gill chloride cell hyperplasia rapidly increased together with increase of salinity, and the number of chloride cells in gill lamellae rapidly increased in response to high external NaCl concentrations. The ultrastructure by scanning electron microscope(SEM) indicated that the gill secondary lamella of tilapia(Oreochromis niloticus) exposed to seawater, were characterized by rough convoluted surfaces during the adaptation. Transmission electron microscopy(TEM) indicated that mitochondria in chloride cells exposed to seawater, were both large and elongate and contained well-developed cristae. TEM also showed the increased chloride cells exposed to seawater. The presence of two mitochondria-rich cell types is discussed with regard to their possible role in the hypoosmoregulatory changes which occur during seawater-adaptation. Most Oreochromis niloticus adapted in seawater had an occasional glomerulus completely filling Bowman's capsule in kidney, and glomerular shrinkage was occurred higher in kidney tissues of individuals living in 10%o, 20%o, 30%o of seawater than in those living in 0%o of freshwater, and blood congestion was occurred severer in kidney tissues of individuals living 20%o, 30%o of seawater than in those living in 10%o of seawater. There were decreases in the glomerular area and the nuclear area in the main segments of the nephron, and that the nuclear areas of the nephron cells in seawater-adapted tilapia were of smaller size than those from freshwater-adapted fish. Our findings demonstrated that Oreochromis niloticus tolerated moderately saline environment and the increased body weight living in 30%o was relatively higher than that living in 10%o in spite of histopathological changes.

• Korean Journal of Veterinary Research
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• v.21 no.2
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• pp.105-112
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• 1981

The morphological and structural studios of Anisakinae larva has been carried out since Sept. of 1980. The larva were collected from naturally infested eleven swine of 1,531 examined at Kwang-Ju abattoir and from marine fishes, Somber japonicus, bought at Kwang-Ju fish market. The results observed were as follow : 1. Anisakis larva found in the stomach wall and on the surface of the mucosa were more or less degenerated. According to the progress of degeneration, the cross sections showed varied structures (Fig. 6, 7). 2. Size of the larva both from swine and fishes were measured respectively in average(mm); 18.0 and 18.7 in body length, 0.30 and 0.41 in body width, 1.64 and 1.68 in esophagus(muscular-part), 0.56 and 0.67 in ventriculus (glandular part), and 0.13 and 0.12 in tail. It was notable that body length of the larva in this present data, 18.0mm and 1.87mm, were shorter than those in previous dada, 24.3mm from human cases and 28.4mm from, however, the present data were almost similar to the data, 1.75mm, from swine case. 3. The Boring tooth, Mucron, long ventriculus and short round tail were observed in the larva of this present study. These structures were differentiated from Anisakis type II larvae which was provided with short ventriculus, and conical and tapering tall without mucron. 4. The ventricular appendix and intestnal caecum were not present in the larva. These might be differentiated from other Anisakidae larva such as Terranova larvae, Contracaecum larvae, Raphidascaris larvae and Thynnascaris larvae. 5. The findings through the histological observation were a pair of Y-shaped or butterfly-shaped lateal chords, ventral and dorsal chords, excretory(Renette) cell, high columnar epithelial cells of digestive tract and muscle cells. These morphological characteristics revealed varied features in the structures in the degenerative degree of the larva in the stomach wall. 6. The above-mentioned characteristics of the larva observed could be indentified as Anisakis type I larvae. 7. The reports on natural infestation of domestic animal with Anisakis type I larvae were two swine cases in Korea and Japan respectively, On the other hand two human cases of the larva were reported in Korea and more than one thousand cases in Japan. In Twiwan no reports of human and domestic animal cases could be found.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.27 no.3
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• pp.226-237
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• 2005

Purpose : Extensive defect of oral and maxillofacial area is usually reconstructed with composite flap including skin paddle. However, if the defects are lined with only skin components, the mucosa's role in mastication and texture are not restored. Furthermore, stiffness and hair-growing prevent denture rehabilitation and good oral hygiene. This study was performed to overcome the disadvantages of composite soft tissue flaps including the skin and to make a model for myo-mucosal flaps. Materials and methods : Buccal mucosa sized $0.5\times1.0\;cm^2$ from New Zealand rabbit (around 1.5kg) was harvested and cultivated by the modification of Rheinwald and Green's keratinocyte culture method. Cultured mucosa was grafted on the fascia of latismus dorsi as form of mucosal sheet. After 7, 10, 14 days, the myomucosal flap was excised and evaluated under light microscope with H & E and immunohistochemical staining. As control group, harvested buccal mucosa from rabbit was transplanted to gracilis muscle(n=6). Results : From 7 days after prelamination, the basal layer of the grafted mucosa resembled that of normal mucosa. As control group, transplanted mucosa had original shape but there's slight inflammatory reaction. Prelaminated mucosa has 19.8$\pm$4.59 cell layers and some samples have more than 20 layers. The expression rate of PCNA was relatively strong (42.9%$\pm$14.1) at the basal layer of grafted mucosa and the laminin was found at the basal layer. On the contrary, prelaminated mucosa at 10 days showed moderate expression rate of PCNA(32.4%$\pm$4.62). We found the mucosal layer was somehow disappeared and there is strong inflammatory reaction. After 14 days prelamination, the grafted oral keratinocytes were almost disappeared and expression of PCNA was not observed. Conclusion : We can make 75 fold large mucosal($3850mm^2$) sheet from small samples of mucosa $(50mm^2)$. Epithelial sheet that grafted on the fascia of muscle underwent differentiation and proliferation. But after 10, 14 days, there was strong inflammatory reaction and the grafted mucosa was destroyed from surface layer. In rabbit model, transfer of fascio-mucosal flap should be done from 7 to 10 days after prelamination.