Background: Cellular remnants in the bioprosthetic heart valve are known to be related to a host's immunologic response and they can form the nidus for calcification. The extracellular matrix of the decellularized valve tissue can also be used as a biological scaffold for cell attachment, endothelialization and tissue reconstitution. Thus, decellularization is the most important part in making a bioprosthetic valve and biological caffold. Many protocols and agents have been suggested for decellularization, yet there ave been few reports about the effect of a treatment with hypotonic solution prior to chemical or enzymatic treatment. This study investigated the effect of a treatment with hypotonic solution and the appropriate environments such as temperature, the treatment duration and the concentration of sodium dodecylsulfate (SDS) for achieving proper decellularization. Material and Method: Porcine aortic valves were decellularized with odium dodecylsulfate at various concentrations (0.25%, 0.5%), time durations (6, 12, 24 hours) and temperatures ($4^{\circ}C$, $20^{\circ}C$)(Group B). Same the number of porcine aortic valves (group A) was treated with hypotonic solution prior to SDS treatment at the same conditions. The duration of exposure to the hypotonic solution was 4, 7 and 14 hours and he temperature was $4^{\circ}C$ and $20^{\circ}C$, respectively. The degree of decellularization was analyzed by performing hematoxylin and eosin staining. Result: There were no differences in the degree of decellularization between the two concentrations (0.25% 0.5%) of SDS. Twenty four hours treatment with SDS revealed the best decellularization effect for both roups A and B at the temperature of $4^{\circ}C$, but there was no differences between the roups at $20^{\circ}C$. Treatment with hypotonic solution (group A) showed a better ecellularization effect at all the matched conditions. Fourteen hours treatment at $4^{\circ}C$ ith ypotonic solution prior to 80S treatment revealed the best decellularization effect. The treatment with hypotonic solution at $20^{\circ}C$ revealed a good decellularization effect, but his showed significant extracellular matrix destruction. Conclusion: The exposure of porcine heart valves to hypotonic solution prior to SDS treatment is highly effective for achieving decellularization. Osmotic treatment with hypotonic solution should be considered or achieving decellularization of porcine aortic valves. Further study should be carried out to see whether the treatment with hypotonic solution could reduce the exposure duration and concentration of chemical detergents, and also to evaluate how the structure of the extracellular matrix of the porcine valve is affected by the exposure to hypotonic solution.
Kim, Yong Hyun;Yoon, Hyoung Kyu;Kim, Chi Hong;Ahn, Joong Hyun;Kwon, Soon Seog;Kim, Young Kyoon;Kim, Kwan Hyoung;Moon, Hwa Sik;Park, Sung Hak;Song, Jeong Sup;Cho, Kyung Sook
Tuberculosis and Respiratory Diseases
/
v.58
no.6
/
pp.590-599
/
2005
Object : Cigarette smoking is a major cause of mucus hypersecretion, which is a pathophysiological feature of many inflammatory airway diseases. Mucins, which are an important part of the airway mucus, are synthesized from the Muc gene in airway epithelial cells. However, the signaling pathways for cigarette smoke-induced mucin synthesis are unknown. The aim of this study was to determine the signal pathway for smoking induced Muc5ac gene expression. Methods : A549 cells were cultured and transiently transfected with the Muc5ac promoter fragment. These cells were stimulated with 5% cigarette smoke extract (CSE) alone or with CSE after a pretreatment with various signal transduction pathway inhibitors (AG1478, PD98059 and SB203580). The Muc5ac promoter activity was examined using the luciferase reporter system, and the level of phosphorylated EGFR, ERK1/2, p38 MAPK and JNK were all examined using Western blot analysis. Muc5ac mRNA expression was also examined using reverse transcriptase polymerase chain reactions (RT-PCR). Results : 1. The peak level of luciferase activity of the Muc5ac promoter was observed at 5% concentration and after 3 hours of incubation with the CSE. The level of EGFR phosphorylation and the luciferase activity of the transfected cells caused by the CSE were significantly suppressed by AG1478 or PD98059 (P<0.01). 2. CSE phosphorylated ERK1/2 or p38 MAPK but not JNK. The Muc5ac mRNA expression level was increased by the CSE but that was suppressed by PD98059 or AG1478. 3. The CSE-induced phosphorylation of ERK1/2 was blocked by PD98059 and that of p38 MAPK was blocked by either PD98059 or SB203580. Either PD98059 or SB203580 suppressed the luciferase activity of the transfected cells (P<0.0001). Conclusion : The Muc5ac mRNA expression level was increased by the CSE. The increased CSE-induced transcriptional activity was mediated via EGF receptor activation, which led to ERK1/2 and p38 MAPK phosphorylation.
Han, Seon-Sook;Sung, Ji Hyun;Lee, Mi-Eun;Lee, Seung-Joon;Lee, Sung Joon;Kim, Woo Jin
Tuberculosis and Respiratory Diseases
/
v.63
no.3
/
pp.235-241
/
2007
Background: Airway mucus hypersecretion plays an important role in the pathogenesis of asthma, and is associated with the induction of MUC5AC expression in airway secretion. The MUC5AC gene is highly polymorphic; however, there are few studies about the association between the polymorphisms of the MUC5AC gene and asthma susceptibility or asthma phenotypes. We have investigated the association of MUC5AC promoter polymorphisms with the risk of asthma and asthma phenotypes. Methods: We determined the genotypes of the MUC5AC promoter (-1274G>A) in 78 asthma patients and in 78 age, sex-matched control individuals in the Korean population. Genomic DNAs from blood were analyzed by PCR and RFLP (restriction fragment length polymorphism) determination. We examined $FEV_1$, total eosinophil count, serum IgE level, $PC_{20}$ and the presence of atopy (by a skin test) in asthma patients. Results: The mean age of the patients was $47.7{\pm}16.1$ years and 38.5% were men, and the mean $FEV_1$ was $84.4{\pm}22.3%$ of predicted in the asthma patients. The -1274G>A polymorphism of the MUC5AC promoter in asthma patients was not significantly different as compared with normal individuals (GG 57.7%, AG 34.6% and AA 7.7% in asthma patients vs. GG 56.4%, AG 38.5% and AA 5.1% in control subject, p = 0.752, Cod). Several clinical parameters in asthma patients such as $FEV_1$, total eosinophil count, serum IgE level, $PC_{20}$ and the presence of atopy, were not associated with the -1274G>A polymorphism of the MUC5AC promoter. Conclusion: The -1274G>A single nucleotide polymorphism (SNP) frequency of the MUC5AC promoter was not associated with asthma in a Korean population.
Purpose: Autoimmune hepatitis is a chronic inflammatory liver disease with unknown cause that is characterized by liver histology, circulating autoantibodies and increased levels of immunoglobulin G. Only sporadic reports are available on autoimmune hepatitis in children. The aim of this study was to evaluate the clinical, biochemical, and histological features, and the long-term outcome of autoimmune hepatitis in Korean children. Methods: We reviewed the medical records of 14 children diagnosed as having autoimmune hepatitis at Seoul National University Children's Hospital from 1990 to 2004, and analyzed clinical, biochemical, and histological features, and clinical outcomes. Results: Mean age at diagnosis was 9 years and 11 of the 14 children were female. Six children presented with acute hepatitis-like manifestations. Jaundice and fatigue were the most common symptoms. Other autoimmune diseases accompanied in 6 children. Anti-nuclear antibody was detected in 13 patients and anti-smooth muscle antibody was positive in 8. All 14 patients were type 1 autoimmune hepatitis. The main histologic findings were interface hepatitis, rosette formation, and cirrhosis. Clinical and biochemical features were improved in six patients treated with ursodeoxycholic acid. Eight patients were treated with corticosteroid alone or in combination with azathioprine and five of them are in biochemical remission. Conclusion: Autoimmune hepatitis is an inflammatory liver disease, which has a favorable long-term outcome if it is diagnosed and treated promptly. Therefore, autoimmune hepatitis should be suspected in children with chronic hepatitis of unknown etiology, especially in female patients who show hypergammaglobulinemia or some clinical features of autoimmune disease.
Background : So far, there have been numerous reports on organ damage due to cryptococcosis, however, cases of lung localization have been infrequently reported. Recently pulmonary cryptococcosis has been reported more frequently than before due to enhanced diagnostic techniques and increased underlying diseases. Method : The author, therefore, analyzed the clinical manifestations of 5 cryptococcosis cases that we experienced at Hanyang University Hospital from 1985 to 1996 and 9 cases reported in Korea from 1984 and 1996 retrospectively. The following results were obtained. Results : Cryptococcosis occurred frequently over sixth decade and the male to female ratio was 3.6 : 1. Underlying diseases included acute rejection after kidney transplantation, rheumatoid arthritis, autoimmune hepatitis, diabetes mellitus and state of bilateral adrenalectomy. Remaining 8 cases had no evidence of an underlying disease. Because the symptoms were subacute & nonspecific, and not improved by conventional antibiotics, 6 patients of 14 pulmonary cryptococcosis patients were treated as pulmonary tuberculosis before correct diagnosis was made. There were three asymptomatic cases. According to the results of CXR, solitary alveolar consolidation was the most common finding(8 cases) followed by diffuse infiltration(5 cases). It also showed pleural effusion, hilar lymphadenopathy and cavity formation that was rarely reported in world literature. The diagnasis was made through fine needle aspiration biopsy in 10 cases, open thoracotomy in 2 cases, transbronchial lung biopsy in 1 case. and thoracentesis with pleural biopsy in 1 case. Only one case showed positive result in sputum stain and culture, serum latex agglutination test for cryptococcus neoformans. Treatment modalities were various such as fluconazole, amphotericin B, flucytosine, ketoconazole, surgery and it's combination. After 1990 year, there was a trend that fluconazole or ketoconazole are more used than other therapeutic modalities. Conclusion : Because the symptoms are subacute & nonspecific and not improved by conventional antibiotics, pulmonary cryptococcosis is likely to misdiagnosis as pulmonary tuberculosis in Korea. Because the diagnosic yield of sputum stain, culture and serologic test for pulmonary cryptococcosis is low, histologic diagnosis is need in most pulmonary cryptococcosis.
Kim, Yeon Sao;Kim, Seong Min;Kim, Jin Ho;Lee, Kyung Sang;Yang, Suck Chul;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo;Lee, Jung Hee;Choi, Yo Won;Jean, Seok Chol;Kim, Young Tae
Tuberculosis and Respiratory Diseases
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v.43
no.4
/
pp.571-578
/
1996
Background : Long abscess is an accumulation of pus within a destroyed portion of the lung. Antibiotic therapy and postural drainge has proven to be an effective method of treatment for the majority of patients with pyogenic lung abscess. When medical therapy fails, thoracotomy and pulmonary resection are the current therapies. empyema is pus in the pleural space, and this term is deserved for effusions on which the Gram stain of the pleural fluid is positive. Initially, such collection may be drained via chest tribe. Recently, in patients who are judged to be unsuitable for surgery are in poor condition, percutaneous drainage using pig-tail catheter has been performed. We report out experience with 10 cases of lung abscess and 23 cases of empyema who were treated by percutaneous pit-tail catheter drainage. Subjects and Methods : Our study included 10 patients with lung abscess and 23 patients with empyema who were treated by percutaneous pig-tail catheter drainage, from January, 1990, to May, 1996, at Hanyang University then a pig-tail catheter was inserted into the abscess or the site of empyema under fluoroscopic and ultrasonograpic guidance. Following aspiration, the catheter was sutured into the skin, and connected to the suction tip. Catheter drainage was discontinued when the abscess of empyema was resolved in radiologically and clinically. Results : There were 2 cases of lung abscess caused by Staphylococcus aureus and Klebsiella pneumoniae and 14 cases of empyema caused by M. tuberculosis. The others were unknown. The duration of drainage was 1-2 weeks in 7 cases of lung abscess and 14 cases of empyema. In the 29 of 33 patients, percutaneous drainage were carried out successfully 20 of the 29 Gases rapidly improved. Conclusion : Percutaneous drainge is effective and relatively saute for management of lung abscess or empyema refractory to medical therapy or poor candidates for surgical treatment.
Cancer is a complex disease heterogeneously composed of various types of cells including cancer stem-like cells responsible for relapse and chemoresistance in the tumor microenvironment. The conventional two-dimensional cell culture-based platform has critical limitations for representing the heterogeneity of cancer cells in the three-dimensional tumor niche in vivo. To overcome this insufficiency, three-dimensional cell culture methods in a scaffold-dependent or -free physical environment have been developed. In this study, we improved and simplified the HCT-8 colon cancer cell-based spheroid culture protocol and evaluated the relationship between cancer stemness and responses of chemosensitivity to 5- Fluorouracil (5-FU), a representative anticancer agent against colon cancer. Supplementation with defined growth factors in the medium and the culture dish of the regular surface with low attachment were required for the formation of constant-sized spheroids containing $CD44^+$ and $CD133^+$ colon cancer stem cells. The chemo-sensitivities of $CD44^+$ cancer stem cells in the spheroids were much lower than those of $CD44^-$ non-stem-like cancer cells, indicating that the chemoresistance to 5-FU is due to the stemness of colon cancer cells. Taken together, the inflammation and oncogenic gut environment-sensitive HCT-8 cell-based colon cancer spheroid culture and comparative evaluation using the simplified model would be an efficient and applicable way to estimate colon cancer stemness and pharmaceutical response to anticancer drugs in the realistic tumor niche.
Extravasation of toxic chemotherapeutic agents cause severe skin ulceration and necrosis which often need secondary surgical intervention. Still, there were not established antidote agent in case of extravasation with mitomycin-c. Dimethyl sulfoxide is known as an effective chemical scavenger of toxic hydroxyl free radical and sodium thiosulfate also was demonstrated significant protector from mitomycin-c induced ulceration by a few experimental studies. Author investigated necrotic area of mitomycin-c injected site and compare to the effectiveness of topical treatment with dimethyl sulfoxide and intradermal injection of sodium thiosulfate according to starting times, forty five mice were divided into 3 groups. Control group(n=5) had no treatment after subcutaneous injection of mitomycin-c. Experimental group I and II were 20 mice treated dimethyl sulfoxide and sodium thiosulfate, respectively. Depending on the starting time of treatment, group I and II were subdivided into 1, 2, 3 and 4 as immediate, 6 hours, 12 hours and 24 hours after mitomycin-c injection. Histologic studies of the necrotic area and survival area after treatment were performed using hematoxylin-eosin staining. The mean necrotic area of group I was significantly decreased depending on the starting time of treatment compared with control group(p<0.01). The results means there was no necrosis area which was treated with topical sodium thiosulfate within 6 hours, and it showed also significant decrease of necrosis area within 24 hours. There was also no necrosis area in group II-1 and significant decrease of necrosis area II-2 and III-3. But, effctiveness of intradermal injection of sodium thiosulfate was not found in group II-4 which was started after 24 hours. Hisotolgic findings showed a bland coagulative necrosis without inflammatory changes and no granulation tissue. The significant difference that cytoplasmic loss of subcutaneous fat and decrease number of hair follicles between two groups resulted from the methods of treatment by topical application and intradermal injection. In conclusion, immediate treatments with topical dimethyl sulfoxide or intradermal injection of sodium thiosulfate signifcantly prevents necrosis by extravasation of mitomycin-c.
Purpose : Bacterial meningitis is a serious disease, especially in the neonatal period, and it carries a significant degree of mortality and morbidity. Group B streptococcus(GBS) is a common cause of neonatal bacterial meningitis. The purpose of this study was to evaluate the clinical manifestations, treatment results and complications of GBS meningitis. Methods : We analyzed 29 cases retrospectively who had been admitted to the pediatric ward or NICU in Asan Medical Center from May 1990 to January 2002. They had proven GBS in culture or latex agglutination test in CSF. Results : The male to female ratio was 1 : 1.9. There were two cases of early onset type and 27 cases of late onset type. All cases had normal birth weight with full term at delivery. The perinatal predisposing factors were premature rupture of membrane(two cases), and maternal colonization(two cases). The most common presenting symptoms were fever and irritability. Associated diseases were GBS sepsis(21 cases). There was relatively high sensitivity to penicillin derivatives. There were abnormal brain CT or MRI findings in 16 cases(64%), such as infarction, encephalomalatic change, effusion, hydrocephalus, hemorrhage and abscess. The intensive care unit admission rate and the incidence of DIC were higher in the group with complications. Two cases were discharged against advice. Conclusion : We recommend early detection and active treatment in Group B streptococcal meningitis to improve the prognosis.
Yoon Sei Chul;Oho Yoon Kyung;Gil Hak Jun;Chung Su Mi;Shinn Kyung Sub;Bahk Yong Whee
Radiation Oncology Journal
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v.5
no.1
/
pp.31-36
/
1987
Radiobiological and clinical evidences indicate that hyperthermia combined with ionizing radiation produces a significant improvement in therapeutic effect of cancer. In general, malignant cells are more sensitive to heat than normal cells in the heat range of $41\~45^{\circ}C$. We report the experiences obtained from 42 patients with advanced malignant neoplasms managed with 2,450MHz microwave-induced local hyperthermia and ionizing radiation at the Department of Radiology, Kangnam St. Mary's Hospital, Catholic University Medical College. A clinical analysis of 42 thermoirradiated patients showed result of 11(26\%),\;15(36\%),\;11(26\%)\;and\;5(12\%)$ patients with complete response (CR), partial response (PR), minor response (MR) and no response (NR), respectively. Histologically there were $17(40.2\%)$ squamous cell carcinomas, $12(28.6\%)$ adenocarcinomas and $6(14.3\%)$ miscellaneous cancers. Eleven patients with CR consisted of five squamous cell carcinomas, five adenocarcinomas, and one chloroma. Among 15 patients with PR were five squamous cell carcinomas, five adenocarcinomas, three unknown primary tumors, and one poorly differentiated, and miscellaneous tumor each.
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