Cyclosporine A(CsA) is a widely used immunosuppressant for transplant patients and is also used for the treatment of a wide variety of systemic diseases with immunologic disorders. However, its use is frequently limited because of complications such as nephrotoxicity or gingival hyperplasia. Although several hypotheses have been postulated for CsA-induced gingival hyperplasia, i.e. various cytokine effects of inflammatory cells, existence of plaque or CsA itself, but its pathogenesis is still unclear. For experimental chronic CsA toxicity, salt depletion has been shown to increased susceptibility of rodents to the effects of CsA, and this maneuver facilitates production of arteriolopathy and interstitial fibrosis in kidney that mimic the changes found in human. The purpose of this study was to evaluate pathogenesis of CsA-induced gingival hyperplasia by comparing changes between CsA administration groups of normal standard diet and those of low salt diet group. Specific pathogen-free, 20 to 25 days old(120 to 150 g), male Fisher-344 rats(KIST, Korea), 120 to 150g of body weight, were assigned to four groups of six animals each after one week of adaptation period for powder food. Group 1 received olive oil($300{\mu}l/g\;of\;diet$) with normal standard diet(0.4% of sodium)(NSD). Group 2 received CsA(Cypol-N, Jonggundang, Korea; $300{\mu}g/g\;of\;diet$) with normal standard diet(NSD+CsA). Group 3 received same amount of olive oil with low salt diet(0.05 % of sodium, Teklad Premier, U.S.A.)(LSD). Group 4 received same dose of CsA with low salt diet(LSD+CsA). Rats were pair fed and were sacrificed after six weeks. Renal histologic lesions associated with CsA, consisted of cortical interstitial fibrosis, tubular atrophy and hyalinization of arterioles and the impairment of renal function including increase of serum creatinine and decrease of glomerular filtration rate was more severe in low salt diet group. These were proved as the results of activated of renin-angiotensin system in the kidney by low salt condition. Meanwhile the degree of gingival hyperplasia at incisor and molar tooth was less severe in low salt diet group compared with normal sodium diet group. Hyperplastic gingiva showed mild epithelial hyperplasia and expanded underlyng stroma which consisted of matrix increasement, capillary proliferation and dilatation. While the number and the activation of fibroblasts were increased, inflammatory cells were rare in the stroma. The immunohistochemistry for TGF-${\beta}_1$ in the kidney and gingiva revealed stronger positive in LSD+CsA in kidney but in gingiva of NSD+CsA. These results suggested followings; Gingival hyperplasia can be developed without inflammatory cells infiltration and seemed not induced by CsA by itself. The major role for gingival hyperplasia by CsA would be the secondary effect of TGF-${\beta}$, which maybe upregulated by CsA administration. Low salt diet can attenuate this hyperplasia perhaps by decreasing the activation of $TGF-{\beta}$.
To compare the morphological and cultural characteristics of Cordyceps militaris, forming conditions of artificial fruiting body production were investigated in this experiment. The stroma shapes of artificial fruiting body were confirmed to club and/or stick. However, both shapes were same color as a orange. Perithecia of stroma was semi-egg shape and was ranged from $30{\sim}90{\times}90{\sim}130\;{\mu}m$ and it's asci was narrowly cylindric. Ascospore of perithecia was belong to filiform and multiseptate. The conidia of C. militaris was centurally grown and shaped with globose, long clavate type, floccose, centrally raised during anamorph stage. When it was cultured between glass and PP bottle, production of artificial fruiting body, pinheading ratio and total yield were higher in PP bottle. The optimum illumination was 1,000 lx for the those production. The culture medium of barley, wheat and hulled rice showed higher artificial fruiting body compared to that of silkworm. Pinheading and yield of it's isolates was decreased at more than three subsequent transculture.
Kim, Yeo-Yae;Lee, Dong-Nyung;Seo, Il-Bock;Kim, Hyung-Jun
The Journal of Korean Obstetrics and Gynecology
/
v.19
no.2
/
pp.162-185
/
2006
Purpose : This study was carried out to investigate the effects of Gamigwichulpajing-Tang(GGT) on the development of experimentally-induced endometriosis in rats. Materials and Methods : Endometriosis was induced in rats by auotransplanting uterine tissue to the peritoneum and devided 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 GGT treated group. GGT(700mg/head) was orally administrated for 15days after operation. Then we measured the body weight, the volumes of endometriotic implants, the weight of uterus and ovary, and investigated the content of cytokines(MCP-1, $TNF-{\alpha}$, $IL-l{\beta}$) in serum and ascites. Histopathology, immunohistochemistry for COX-2, and histochemistry for mast cell in transplanted uterine tissue were performed. Results :- The $volume(mm^3)$ of endometriotic implants in GGT-treated group$(53.50\;{\pm}18.63)$ was significantly decreased(p<<0.01) compared with control group$(404.50{\pm}317.68)$. - The content(pg/ml) of MCP-1 in ascites in GGT-treated group$(4265{\pm}108)$ was significantly decreased(p<<0.001) compared with control group$(8632{\pm}1245)$. - The content(pg/ml) of $TNF-{\alpha}$ in serum in GGT-treated group$(64.5{\pm}21.6)$ was significantly decreased(p<<0.05) compared with control group$(147.1{\pm}78.2)$. - The content(pg/ml)- of $TNF-{\alpha}$ in ascites in GGT-treated group$(738.3{\pm}502.4)$ was significantly decreased(p<<0.05) compared with control group$(1245.2{\pm}362.2)$. - The percentage(%) of positive epithelial layers for COX-2 in GGT-treated group$(22.9{\pm}9.3)$ was significantly decreased(p<<0.001) compared with control group$(50.2{\pm}8.2)$. - The number of mast cells in adjacent tissue of transplanted uterine tissue in GGT-treated group$(61.4{\pm}13.9)$ was significantly decreased(p<<0.001) compared with control group$(109.3{\pm}30.2)$. - The number of mast cells in stroma of transplanted uterine tissue in GGT-treated group$(9.4{\pm}2.7)$ was significantly decreased(p<<0.001) compared with control group$(26.0{\pm}7.7)$. - Histopathologically, proliferation of endomeuiotic epithelia and stroma, and infiltration of inflammatory cells in transplanted uterine tissue of GGT-treated group were weakly observed than those of control group. Conclusion :On the basis of these results, we concluded that Gamigwichulpajing-Tang have inhibiting effects on the development of transplanted uterine tissue. And these effects may be related with decreased Production of MCP-1 and $TNF-{\alpha}$, decreased expression of COX-2, and decreased infiltration of mast cells by administration of Gamigwichulpajing-Tang.
Kim, Chul-Soo;Lee, Dong-Nyung;Seo, Il-Bock;Kim, Hyung-Jun
The Journal of Korean Obstetrics and Gynecology
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v.20
no.1
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pp.61-83
/
2007
Purpose: This study was performed to investigate the effects of Keukhachukeo-Tang (KCT) on the development of experimentally-induced endometriosis in rats. Methods : Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and devided 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 KCT treated group. KCT (1,200 mg/head) was orally administrated for 15 days after operation. Then we measured the body weight, the volumes of endometriotic implants. The weight (body, left uterus and ovaries) and concentrations of cytokines (MCP-1, TNF-${\alpha}$, IL-l${\beta}$) in serum and peritoneal fluid were also measured. Histopathology, immunohistochemistry for COX-2, and histochemistry for mast cells in transplanted uterine tissue were performed. Results : - The volumes(mm$^3$) of endometriotic implants in KCT-treated group (107${\pm}$66) were significantly decreased (p<0.05) compared with control group (405${\pm}$318). - The contents(pg/ml) of MCP-1 in peritoneal fluid in KCT-treated group (6,940${\pm}$893) were significantly decreased (p<0.01) compared with control group (8,632${\pm}$1,245). - The contents(pg/ml) of TNF-${\alpha}$ in peritoneal fluid in KCT-treated group (847${\pm}$330) were significantly decreased (p<0.05) compared with control group (1,245${\pm}$362). - The percentages(%) of positive epithelial layers for COX-2 in KCT-treated group (31${\pm}$10) were significantly decreased (p<0.01) compared with control group (50${\pm}$8). - The numbers of mast cells in adjacent tissue of transplanted uterine tissue in KCT-treated group (69${\pm}$18) were significantly decreased (p<0.01) compared with control group (109${\pm}$30). - The numbers of mast cells in stroma of transplanted uterine tissue in KCT-treated group(16${\pm}$5) were significantly decreased (p<0.01) compared with control group (26${\pm}$8). - Histopathologically, proliferation of endometriotic epithelia and stroma, and infiltration of inflammatory cells in transplanted uterine tissue of KCT-treated group were weakly observed than those of control group. Conclusion : From the above results, Keukhachukeo-Tang (KCT) have inhibiting effects on the development of transplanted uterine tissue. And these effects are related to the decreased concentration of MCP-1 and TNF-${\alpha}$, decreased expression of COX-2, and decreased infiltration of mast cells by administration of Keukhachukeo-Tang.
Two groups of esophagus graft were done in canine esophagus in 34 adult mongrel dogs. For the first group segmental replacement graft was done with fresh autologous pericardium tube, and for the second, patch graft was done utilizing fresh autologous pericardium, fresh homologous pericardium,and dacron piece. All eight dogs in the first segmental replacement graft group died 2 to 5 days after operation with severe empyema caused by anastomosis disruption. Among 26 patch graft dogs 2 died during operation and 7 died 13 to 18 days after operation. For the 17 long-term patch grafted survivors esophagography and postoperative weight check were done. Postoperative stool was collected and examined for dacron patch excretion. One, two, three, and four months postoperative long-term survivors were sacrificed to obtain specimens in each group respectively and the following observations were made.I. Survival; Autologous pericardium patch group showed no mortality but in homologous pericardium and dacron patch group only two thirds were long-term survivors. II. Postoperative swallowing; There was no case which demonstrated postoperative dysphagia. About half of the cases showed postoperative weight increase and in only 3 cases weight decrease followed operation. III. Dacron patch was excreted in the stool 8 to 23 days after operation. Animals which excreted dacron patch up to 9 days after operation all died of empyema due to anastomosis disruption.IV. Postoperative esophagogram; All esophagograms in each group showed no leakage of barium, no passage disturbances and no remarkable stenotic signs.V. Morphological findings; [A] Macroscopical findings; In one month group specimens of each group dense adhesion with surrounding structures was noted and luminal surface was smooth with contraction of the patched area. In two month groups anastomosis sutures were still exposed but patched area showed lesser abnormality. In three to four months groups sutures were covered completely and patched area showed only very slight signs of contraction. [B] Microscopic findings; In one month group luminal surface of the replaced tissue [transplanted tissue] showed almost complete epithelial covering that is composed of several layers of squamous cells with no evidence of keratinization. Basement membrane was also well distinct throughout. Slight to minimal inflammatory cells comprising of large mononuclears, lymphocytes and plasma cells were observed in the subepithelial fibrous stroma consisted entirely of loose fibrous tissue containing many newly formed capillaries and fibroblastic proliferation. Scattered suture granulomas were found, few of which became acutely inflamed. In two months group repairing process progressed with lesser degree of inflammatory cell infiltration and young capillary proliferation. Fibrous tissue was more matured showing even focal collagenization.Suture granuloma persisted but with lesser reactive changes. Epithelial covering was that of a mature non-keratinizing stratified squamous epithelium. In three and four months groups the replaced area showed essentially similar histological findings. However, subepithelial stroma still consisted entirely of connective tissue without evidence of smooth muscle regeneration. In this group, inflammatory cell infiltration was minimal or negligible. Among these patch materials autologous pericardium group showed the most satisfactory repairing process.The above mentioned results may signify the feasibility of autogenous pericardium patch graft in clinical esophageal surgery.
Two groups of esophagus graft were done in canine esophagus in 34 adult mongrel dogs. For the first group segmental replacement graft was done with fresh autologous pericardium tube, and for the second, patch graft was done utilizing fresh autologous pericardium, fresh homologous pericardium,and dacron piece. All eight dogs in the first segmental replacement graft group died 2 to 5 days after operation with severe empyema caused by anastomosis disruption. Among 26 patch graft dogs 2 died during operation and 7 died 13 to 18 days after operation. For the 17 long-term patch grafted survivors esophagography and postoperative weight check were done. Postoperative stool was collected and examined for dacron patch excretion. One, two, three, and four months postoperative long-term survivors were sacrificed to obtain specimens in each group respectively and the following observations were made. I. Survival; Autologous pericardium patch group showed no mortality but in homologous pericardium and dacron patch group only two thirds were long-term survivors. II. Postoperative swallowing; There was no case which demonstrated postoperative dysphagia. About half of the cases showed postoperative weight increase and in only 3 cases weight decrease followed operation. III. Dacron patch was excreted in the stool 8 to 23 days after operation. Animals which excreted dacron patch up to 9 days after operation all died of empyema due to anastomosis disruption. IV. Postoperative esophagogram; All esophagograms in each group showed no leakage of barium, no passage disturbances and no remarkable stenotic signs. V. Morphological findings; [A] Macroscopical findings; In one month group specimens of each group dense adhesion with surrounding structures was noted and luminal surface was smooth with contraction of the patched area. In two month groups anastomosis sutures were still exposed but patched area showed lesser abnormality. In three to four months groups sutures were covered completely and patched area showed only very slight signs of contraction. [B] Microscopic findings; In one month group luminal surface of the replaced tissue [transplanted tissue] showed almost complete epithelial covering that is composed of several layers of squamous cells with no evidence of keratinization. Basement membrane was also well distinct throughout. Slight to minimal inflammatory cells comprising of large mononuclears, lymphocytes and plasma cells were observed in the subepithelial fibrous stroma consisted entirely of loose fibrous tissue containing many newly formed capillaries and fibroblastic proliferation. Scattered suture granulomas were found, few of which became acutely inflamed. In two months group repairing process progressed with lesser degree of inflammatory cell infiltration and young capillary proliferation. Fibrous tissue was more matured showing even focal collagenization. Suture granuloma persisted but with lesser reactive changes. Epithelial covering was that of a mature non-keratinizing stratified squamous epithelium. In three and four months groups the replaced area showed essentially similar histological findings. However, subepithelial stroma still consisted entirely of connective tissue without evidence of smooth muscle regeneration. In this group, inflammatory cell infiltration was minimal or negligible. Among these patch materials autologous pericardium group showed the most satisfactory repairing process. The above mentioned results may signify the feasibility of autogenous pericardium patch graft in clinical esophageal surgery.
Purpose: To better understand the corneal regeneration after alkali burn regarding the initial clinical progression and the therapy, we investigated the changes of the multi factors following chemical injury in cornea. Methods: This study was performed to observation on the healing process of alkali burned cornea in aspect of immunohistochemistry by immunofluorescence or H-E staining and TUNEL assay. Results: The results showed that although a healing process occurred after alkali burn, apoptosis of epithelial, stromal and endothelial cells in the cornea was continuously observed. Neovascularization and expression of ${\alpha}$-smooth muscle actin (${\alpha}$-SMA) from limbus and from injured cornea, respectively, were observed after 3 days of alkali burn. Formation of collagen III in corneal stroma and increased expression of chondroitin sulfate are coincident with expression of ${\alpha}$-SMA and transforming growth factor-${\beta}$ (TGF-${\beta}$). Conclusions: These data suggest that medical treatment within 3 days of alkali burn will be effective to inhibit neovascularization and formation of collagen III and chondroitin sulfate. This study extends our immumohistochemical understanding of healing process in alkali burned cornea, and the results get in this study will be cornerstones in the development of therapeutic agent for accelerating renewal of chemical damaged cornea.
Protease activities were measured in vegetables used as cooking material or a side dish. Proteases of green pepper (Kwari and Chungyang), perilla leaves, soybean sprout and mungbean sprout were showed high activities. Especially, protease in soybean sprout was the highest activity among them. After dialysis, remaining activities in mungbean sprout, green pepper (Kwari and Chungyang) and perilla leaves were 12, 23, 45% and 37%. In the results of thawing after freezing the proteases, remaining activities in sprout of mungbean and soybean were 100% and 65%. Protease in soybean sprout was showed higher activity and stability than others. Proteolytic effects of soybean sprout on myofibrillar and sarcoplasmic protein were showed higher than stroma protein. In SDS-PAGE, myosin heavy chain, actin and tropomyosin were hydrolyzed with increasing time. Protease activity was rapidly increased with increasing meat concentration in a early proteolysis reaction, but was slightly increased in later.
The reaction center Dl protein of photosystem II is the target of photodamage by excess illumination. The Dl protein is damaged by reactive oxygen species generated by photochemical reactions and then degraded by specific proteolytic enzymes. We found that the Dl protein also cross-links with the surrounding polypeptides, such as D2 and CP43 in isolated thylakoids or photosystem II-enriched membranes from spinach under the illumination with strong visible light. The cross-linking was observed in spinach leaf discs as well when they were illuminated at higher temperature (40°C). It was also shown that the cross-linked products are digested efficiently by a protease(s) in the stroma. Thus the cross-linking/digestion processes of the Dl protein seem to comprise a new pathway in the turnover of the photodamaged Dl protein. It should be noted, however, that the cross-linked products of the Dl protein and CP43 induced by endogenous cationic radicals in the donor-side photoinhibition are resistant to proteolytic digestion. Accumulation of these cross-linked products in the thylakoids may lead to the decay of the function of chloroplasts and finally to the death of plant cells. Thus, we suggest that the quality control of photosystem II, especially removal of the cross-linked products of the Dl protein, is crucial for the survival of chloroplasts under the light stress.
This study was designed in order to investigate the effect of copper sulfate to the ultrastructural changes of the hepatocytes in Sprague Dawley rats. The animals were administered with copper sulfate (10mg/kg B.W.), which was dissolved in normal saline. The solution was injected into abdominal cavity every day. The animals were sacrificed at the 6th, 12th, and 24th day from the beginning of administration. The specimens obtained from the liver were observed with electron microscope and significant changes were as follows. 1. A prominent dilatation and disruption of the cisternae of rough endoplasmic reticulum were recognized. Also, the detachment of membrane bound ribosomes was shown. 2. The proliferation of smooth endoplasmic reticulum and the depletion of glycogen particles were noted. 3. The increase of primary lysosomes and autophagic vacuoles was obserbed. 4. The dilatation of mitochondrial cristae was obserbed. And it was irregulary scattered in the stroma of mitochondria. 5. The atrophy of microvilli in the bile canaliculi and space of Disse was prominent. 6. Membrane of hepatocytes was damaged and significant hydrophic degeneration was obserbed in the perisinusoidal regions. 7. The damage of Fat-storing cells was more significant than that of hepatocytes.
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