BACKGROUND: The immortalized human keratinocyte line, HaCaT cells have been widely used as substitutes for normal epidermal keratinocytes. Recently, reconstruction of a skin equivalent using HaCaT cells showed a multilayered epithelium,but somewhat different tissue architecture as compared with normal epidermis. OBJECTIVE: In this study, using HaCaT cells we tried to reconstruct an epidermis resembling more closely to normal epidermis than the previous results. MATERIALS AND METHODS: HaCaT cells were cultured in air-liquid interface on a recently developed dermal substrated in our laboratory, de-epidermized dermis (DED) raised on fibroblast-populated collagen matrix and the result was compared with those on DED or fibroblast-populated collagen matrix alone. RESULTS: HaCaT cells on the new dermal substrate formed a multilayered epithelium with rete ridges, showing rather orderly cellular organization compared with those on fibroblast-populated collagen matrix. However, horny and granular layers were not observed contrary to normal epidermis. Immunohistochemical studies revealed that differentiation markers such as keratin 1, keratin 6 and involucrin showed the similar pattern to those in HaCaT cells cultured on fibroblast-populated collagen matrix. Markers of terminal differentiation, loricrin and filaggrin were not expressed contrary to normal epidermis. CONCLUSION: These results suggest that organotypic culture HaCaT cells on the dermal substrate combines DED with fivroblast-populated collagen matrix results in incomplete differentiation of HaCaT cells contrary to normal keratinocytes.
Antigens, Differentiation ; Collagen* ; Dermis* ; Epidermis ; Epithelium ; Humans ; Keratin-1 ; Keratin-6 ; Keratinocytes ; Skin

BACKGROUND: Melanocytes grown in pure monolayer culure lack many of the cellular interactions that exist in vivo. This can be partially overcome by growing melanocytes together with other epidermal cells in skin equivalent models. OBJECTIVE: The objective of the present study was to grow human melanocytes in human epidermis reconstructed on dermal substrates in vitro and to examine their response to UV radiation. METHODS: The skin equivalents were prepared by seeding cultured human keratinocytes together with cultured human melanocytes(in a ratio of 5%) onto de-epidermized dermis. After 7 days of culture, they were exposed to UVB irradiation(total 150m J/cm over 5days). On day 12 of air exposure the sections of the skin equivalents were prepared for histology. The structure of the skin equivalents was studied following staining with hematoxylin and eosin. Melanocytes were characterized by DOPA staining and by immunohistochemistry. RESULTS: Melanocytes were localized singly within the basal layer of the reconstructs. Melanin was also visible both in the melanocytes and in neighboring keratinocytes. There was an increase in melanocyte size and dendricity following UV irradiation. Melanocytes became positive to staining with HMB-45 antibody following UV irradiation. CONCLUSION: Our results indicate that melanocytes grown in reconstructed human epidermis are functional and capable of responding to UV irradiation.
Dermis ; Dihydroxyphenylalanine ; Eosine Yellowish-(YS) ; Epidermis* ; Hematoxylin ; Humans* ; Immunohistochemistry ; Keratinocytes ; Melanins ; Melanocytes* ; Skin

BACKGROUND: A range of well characterized materials are needed for validating the performance of hepatitis B surface antigen (HBsAg) immunoassays. These materials are purchased currently from overseas manufacturers at a high cost and with limited quantity. This study was conducted to establish an HBsAg low titer performance panel for use as a national standard for validation of HBsAg immunoassays in Korea. METHODS: 476 plasma units reactive on blood donor screening were collected HBsAg was tested using 3 enzyme immunoassays (EIA) and 1 chemiluminescence immunoassay (CIA). Units reactive on the CIA assay or on 2 or more immunoassays were subjected to hepatitis B virus (HBV) DNA quantification, HBV genotyping and subtyping. Units reactive on HBV DNA quantification were confirmed for HBsAg by neutralization. Candidates for the panel were subjected to a collaborative study performed at 7 laboratories using 7 immunoassays. RESULTS: Eleven HBsAg positive units were selected for the low titer performance panel based on HBsAg immunoassay, HBV DNA quantification, HBV genotyping and subtyping results. The range of the HBsAg concentration of the panel members was 0.05~1.28 IU/mL. Two HBsAg negative units were also included as negative controls. CONCLUSION: As a result of this study, a low titer performance panel [KFDA standard (08/028); HBsAg low titer performance panel (BTRL HBV/LP)] for validation of HBsAg immunoassays has been established as a Korean national standard. Use of this panel will improve performance assessment of HBsAg immunoassays. Because the performance of immunoassays cannot be assessed properly with a limited number of panels, continuous efforts are needed to develop a range of performance panels.
Blood Donors ; DNA ; Hepatitis ; Hepatitis B ; Hepatitis B Surface Antigens ; Hepatitis B virus ; Humans ; Immunoassay ; Immunoenzyme Techniques ; Luminescence ; Mass Screening ; Plasma

Extrarenal Wilms' tumor is extremely rare and occurs predominantly in children. There are less than 50 well-documented cases of extrarenal Wilms' tumor in the literature. Herein, a case of extrarenal Wilms' tumor, whose mass was found in the inguinal region during an operation for the cryptorchidism, is reported in a 2 year old child with a brief review of literature. This case report illustrates the importance of pathologic examination of similar soft masses, found incidentally during operations.
Child ; Child, Preschool ; Cryptorchidism ; Humans ; Male ; Wilms Tumor*

The effect of alloxan-diabetic rat fed with normal, high fat, low protein and high protein diets on the rate of urea production and the activities of enzymes associated with the urea cycle (ornithine transcarbamoylase, E.C. 2.1.3.3, OTC; arginase, E.C. 3.5.5.1) have been studied in intact and isolated perfused liver. The amount of urea excretion was the highest in the high protein diet group. When each diet group was treated with alloxan, total urea excretion showed little differences between each diet group and its corresponding control group with the exception being in the normal diet group. However, the enzyme activity of OTC was increased significantly by alloxan treatment in low and high protein diet groups as compared to corresponding control groups. Similar results were obtained in arginase activity, although the magnitude of the change was less marked. In liver perfusion experiments on rats treated with alloxan, the amount of urea production and changes in OTC and arginase activity were very similar with those in the intact liver. These results suggest that alloxan treatment in normal diet group causes an increase in urea excretion both in intact and perfused liver regardless of changes in enzyme activities and total urea excretion, and enzyme activities are affected by changes in dietary components but the changes of enzyme activities may not correlate with total urea excretion.
Alloxan ; Animal ; Diabetes Mellitus, Experimental/metabolism* ; Dietary Fats/pharmacology* ; Dietary Proteins/pharmacology* ; In Vitro ; Liver/metabolism* ; Male ; Perfusion ; Rats ; Urea/metabolism* ; Urea/urine

Since the 1970s, the average life expectancy of Koreans has been increasing. Our concerns are not simply about longevity, but also about disability-free life. lately, Sasang constitutional medicine has attracted public attention, especially with respect to the elderly. The purpose of this study was to investigate the food habits, nutrients intake, and disease distribution among the elderly, according to their Sasang constitutions. The results are as follow : The distribution of constitutions of the subjects were Tae-eum 48.8%, So-yang 27.4%, So-eum 22.6%, and So-yang + So-eum 1.2%. In both elderly men and women, body weight, percent ideal body weight, and diastolic pressure of the Tae-eum were significantly higher than those of other constitutions. In both men and women, the level of hemoglobin in the Tae-eum was significantly higher than those of other constitutions. The level of LDL-cholesterol was higher, and the level of HDL-cholesterol was lower in the So-eum elderly women. Food habits of the So-eum were the most irregular, and nutrients intake of the So-eum was lower than that of other constitutions in both elderly men and women. In elderly men, the incidence of hypertension was the highest in the Tae-eum, and the incidence of gastritis was the highest in both So-yang and So-eum. In elderly women, the incidence of fatty liver was the highest in the Tae-eum and So-yang, and gastritis and hypertension were the highest in the So-eum. In the elderly, obese and anemic persons were significantly more in the Tae-eum and the So-eum, respectively. The results of this study showed that there were significant differences of physical, characeristics, food, habits, and distributions of disease for the subjects according to each Sasang constitutions. Intake of nutrient was poor, regardless of Sasang constitutions, in both men and women.
Aged* ; Blood Pressure ; Body Weight ; Constitution and Bylaws* ; Fatty Liver ; Female ; Food Habits* ; Gastritis ; Humans ; Hypertension ; Ideal Body Weight ; Incidence ; Life Expectancy ; Longevity ; Male

Linear psoriasis is a rare form of psoriasis and is characterized by the linear distribution of the psoriatic lesions. It should be differentiated clinically and histopathologically from inflammatory linear verrucous epidermal nevus(ILVEN) or psoriasis occuring as part of the Koebner phenomenon. A 41-year-old female is described, who had developed linearly arranged psoriatic lesions in the right lower extremity over the last 10 years. Not only pathological and immunohistochemical studies but also the clinical response to therapies reveal that this is the first case of linear psoriasis in Korea.
Adult ; Female ; Humans ; Korea ; Lower Extremity ; Psoriasis*

BACKGROUND: Neurofibiomat,osis type 1(NF-1) is a multisystemic disorder of genetic ori gin, affecting one in every 3000 to 4000 people. It is clinically important in the aspect of dermatology, pediatrics, orthopedic surgery, neurology, neurosurgery and ophthalmology. OBJECTIVE: The purpore of this study was to elucidate the clinical characteristics of NF-1 in Korean people. METHODS: We carried out a retrospective study on 112 patients which were compatible to the diagnostic criteria of Riccardi and Neurofibromatosis Conference Statement. The results were compared with other western studies. RESULTS: The age of onset, sex ratio, family history of neurofibromatosis, and clinica features of cafe-au-lait spot, neurofibroma, and axillary freckinings did not differed from western countries. However, some characterist,ics of NF 1(e.g. Lisch nodule) were not as sessed in the most of the cases and incomplete evaluations of the systemic diseases wen found. CONCLUSION: In this study t.he clinial features of NF-1 did not differ from western coun tries in many aspects. A more intensive evaluation of patient,s status is needed to manag; NF-1 patients appropritely.
Age of Onset ; Cafe-au-Lait Spots ; Dermatology ; Humans ; Neurofibroma ; Neurofibromatoses* ; Neurofibromatosis 1* ; Neurology ; Neurosurgery ; Ophthalmology ; Orthopedics ; Pediatrics ; Retrospective Studies ; Sex Ratio

BACKGROUND: A number of in vitro skin models have been developed for the purpose of the screening of cosmetics, pharmaceuticals, and environmental chemicals. To mimic the skin in vivo, a model should resemble morphologically and biochemically the parent, tissue. OBJECTIVE: The purpos of this study is to study the differentiation and organization of the artificial epidermis in comparsion with epidermis in vivo based on the expression of epidermal protein antigens and basement membrane components. METHODS: Human keratinocytes were cultured on deepidermidized dermis (RE-DED) or on fibroblast-populated collag-,n matrix (LSE). After 10 days culture, the sections of RE-DED and LSE were stained with hematoxylin and eosin. An immunohistochemical study was also performed with the sections of RE-DED and LSE using antibodies recognizing proliferating cell nuclear antigens (PCNA), epidermal growth factor receptor (EGFR), keratin 1, involucrin, filaggrin, loricrin, keratin 13, type IV collagen, and laminin. RESULTS: In both culture systems(RE-DED and LSE) a multilayered epidermis with a horny layer was observed. In the human epidermis reconstructed by both culture systems, differentiation markers appeared but with a topography slightly different from that of epidermis in vivo, and components of the basement membrane was also expressed. CONCLUSION: Our findings suggest the epidermis obtained in both culture systems(RE-DED and LSE) resembled in vivo epidermis morphologically and biochemically, although it was not the same.
Antibodies ; Antigens, Differentiation ; Basement Membrane* ; Collagen Type IV ; Dermis ; Eosine Yellowish-(YS) ; Epidermis* ; Hematoxylin ; Humans* ; Keratin-1 ; Keratin-13 ; Keratinocytes ; Laminin ; Mass Screening ; Parents ; Proliferating Cell Nuclear Antigen ; Receptor, Epidermal Growth Factor ; Skin

BACKGROUND: A number of in vitro skin models have been developed for the purpose of the screening of cosmetics, pharmaceuticals, and environmental chemicals. To mimic the skin in vivo, a model should resemble morphologically and biochemically the parent, tissue. OBJECTIVE: The purpos of this study is to study the differentiation and organization of the artificial epidermis in comparsion with epidermis in vivo based on the expression of epidermal protein antigens and basement membrane components. METHODS: Human keratinocytes were cultured on deepidermidized dermis (RE-DED) or on fibroblast-populated collag-,n matrix (LSE). After 10 days culture, the sections of RE-DED and LSE were stained with hematoxylin and eosin. An immunohistochemical study was also performed with the sections of RE-DED and LSE using antibodies recognizing proliferating cell nuclear antigens (PCNA), epidermal growth factor receptor (EGFR), keratin 1, involucrin, filaggrin, loricrin, keratin 13, type IV collagen, and laminin. RESULTS: In both culture systems(RE-DED and LSE) a multilayered epidermis with a horny layer was observed. In the human epidermis reconstructed by both culture systems, differentiation markers appeared but with a topography slightly different from that of epidermis in vivo, and components of the basement membrane was also expressed. CONCLUSION: Our findings suggest the epidermis obtained in both culture systems(RE-DED and LSE) resembled in vivo epidermis morphologically and biochemically, although it was not the same.
Antibodies ; Antigens, Differentiation ; Basement Membrane* ; Collagen Type IV ; Dermis ; Eosine Yellowish-(YS) ; Epidermis* ; Hematoxylin ; Humans* ; Keratin-1 ; Keratin-13 ; Keratinocytes ; Laminin ; Mass Screening ; Parents ; Proliferating Cell Nuclear Antigen ; Receptor, Epidermal Growth Factor ; Skin