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

Adult ; Female ; Humans ; Keratin-1 ; genetics ; Keratin-9 ; genetics ; Keratoderma, Palmoplantar ; genetics ; Male ; Middle Aged ; Pedigree

BACKGROUND: Calcipotriol(MC903), a new vitamin D(3) analogue, has been reported to be effective in the treatment of patients with psoriasis. OBJECTIVE: The purpose of this study is to examine the effects of calcipotriol on proliferation and differentiation of the keratinocytes in monolayer cultures and three-dimensional cultures. METHODS: Using moriolayer cultures, we examined morphological changes of keratinocytes and performed [(3)H]thymidine incorporation after calcipotriol was added into the medium. Using three dimensional cultures, we performed two experiments: one with cultures treated with calcipotriol immediately after the keratinocytes had been exposed to the air and another set of cultures treated with calcipotriol after three dimensional morphogenesis of the keratinocytes. We examined morphological changes of keraitinocytes and performed a immunohistochemical study for proliferation differentiation markers RESULTS: In monolayer cultures, at calcipotriol concentrations of 10(-9)M-10(-6)M, keratinocytes became larger, more irregular, and flattened in a dose-dependent manner. At 10(-9)M-10(-6)M, [3Hl thymidine incorporatiorn was decreased dose-dependently as compared to the control culture. In the first experiment using three-dimensional cultures, at 10(-9)M-10(-6)M, total epidermal layers were thinned. This was associated with thinnings of nucleated and horny layers in a dose dependent manner. In the seconcd experiment using three-dimensional cultures, at 10(-8)M-10(-6)M, nucleated layers were thinned in a dose dependent manner, but the horny layer was slightly thickened, as compared to the control culture. Immunohistochemical studies showed a reduction of differentiation markers such as keratin 1, involucrin, filaggrin, loricrin consistent with a thinning of nucleated layers in the epidermal architecture in both experiments. In the basal layer, at 10(-9)M-10(-6), PCNA-positive cells were and BrdU-positive cells were decreased dose-dependently as compared to the control culture. CONCLUSIONS: In this study, we demonstrated that at 10(-9)M-10(-6) calcipotriol inhibited keratinocytes proliferation and stimulated keratinocytes differentiation in a dose-dependent manner.
Antigens, Differentiation ; Humans* ; Keratin-1 ; Keratinocytes* ; Morphogenesis ; Psoriasis ; Thymidine ; Vitamins

Roughly 1% of mature cystic teratomas undergo malignant transformation. In particular, cutaneous-type adnexal neoplasms may occur in mature cystic teratomas. Sebaceous carcinomas, which arise from mature cystic teratomas, have rarely been observed, with only seven cases previously reported. Here, we present a case of a 69-year-old female who had pelvic pain for two weeks and who subsequently underwent bilateral salpingo-oophorectomy and hysterectomy. Her left ovary showed a unilocular cyst, measuring 22.0 cm in diameter, filled with sebaceous material and a few hairs. A luminally-protruding solid mass measuring 4.0 cm in diameter was also noted. Microscopic findings revealed lobular or diffusely arranged basophilic, atypical sebaceous cells connected to a typical mature cystic teratoma. Tumor cells demonstrated positive immunoreactivity for high molecular weight cytokeratin, cytokeratin 7, cytokeratin 19, epithelial membrane antigen, and carcinoembryonic antigen. Here, we present a case of sebaceous carcinoma arising from a mature cystic teratoma along with a review of previously published reports.
Basophils ; Carcinoembryonic Antigen ; Female ; Hair ; Humans ; Hysterectomy ; Keratin-19 ; Keratin-7 ; Keratins ; Molecular Weight ; Mucin-1 ; Ovary ; Pelvic Pain ; Teratoma

BACKGROUND: Due to advancements in treatment of metastatic and advanced renal cell carcinoma (RCC), it has become increasingly important to diagnose metastatic RCC and the specific subtype. In this study, we investigated the diverse histologic features of metastatic clear cell renal cell carcinoma (CCRCC) cases in comparison with corresponding primary lesions. METHODS: We identified 119 metastatic CCRCC cases from 81 corresponding primary lesions diagnosed between 1995 and 2010 and evaluated the diverse histologic and immunohistochemical features of these lesions. RESULTS: A total of 44 primary lesions (54.3%) had a non-clear cell component in addition to a typical clear cell component. Of the 119 metastatic lesions, 63 lesions (52.9%) contained a non-clear cell component, and 29 metastatic lesions were composed of a non-clear cell component only. Rhabdoid features were the most frequent non-clear cell histology among the metastatic lesions. Metastatic CCRCCs mainly showed positive CD10 and epithelial membrane antigen staining and negative cytokeratin 7 staining. CONCLUSIONS: Metastatic CCRCC commonly showed a variety of histologic features. If there is a difficulty to diagnose metastatic CCRCC due to a variety of histologic features or small biopsy specimen, histologic review of the primary lesion and immunohistochemical analysis can help determine the correct diagnosis.
Biopsy ; Carcinoma, Renal Cell* ; Cellular Structures ; Immunohistochemistry ; Keratin-7 ; Mucin-1 ; Neoplasm Metastasis

OBJECTIVETo map and identify the disease gene for the epidermolytic palmoplantar keratoderma (EPPK) in a Uighur family of China.

METHODSBlood samples were collected and genomic DNA was extracted from 48 members of the Xinjiang Uighur family. Six microsatellite repeat sequences on chromosome region 17q12-q21 and 12q13 were selected based on the two known candidate genes KRT9 and KRT1. Two-point linkage analysis and haplotype analysis were performed. Exons and their flanking intronic sequence of the KRT9 gene were amplified by polymerase chain reaction (PCR) and sequenced.

RESULTSData from the marker D17S1787 suggested linkage and yielded a Lod score of 8.65 at theta=0 by using MLINK software. Genotypes and haplotypes were acquired. The disease gene of the EPPK family is located between markers 17/TG/36620115 and D17S846. Chromosome 12q13 region was excluded with the negative Lod score obtained in marker D12S96 (Lod=-infinity at theta=0). No pathogenic mutation was detected in the KRT9 gene.

CONCLUSIONThe disease gene of the EPPK family is located on chromosome region 17q21.2. The keratin 9 gene might not be the disease gene.

China ; Chromosomes, Human, Pair 17 ; genetics ; Female ; Humans ; Keratin-1 ; genetics ; Keratin-9 ; genetics ; Keratoderma, Palmoplantar, Epidermolytic ; ethnology ; genetics ; Male ; Microsatellite Repeats ; Mutation ; Pedigree

Cutaneous metastasis from gastric adenocarcinoma is uncommon, and the eyelid is a rare metastatic site. Three patterns of clinical presentation of eyelid metastasis have been described: nodular, infiltrative, and ulcerated. The infiltrative pattern, also known as an inflammatory diffuse pattern or mask-like metastasis, can be easily misdiagnosed as cellulitis or contact dermatitis. Here, we report a case of gastric adenocarcinoma in a 75-year-old man who presented with a localized erythematous plaque on his eyelid that developed four months earlier. The patient had been treated with an antimicrobial agent owing to suspicion of preseptal cellulitis. Gastric adenocarcinoma metastasis was diagnosed on the basis of histopathological examination and immunophenotyping (i.e., cytoplasmic epithelial membrane antigen, cytokeratin- 7, cytokeratin-20, and carcinoembryonic antigen). For patients with malignant neoplasms, persistent skin lesions similar to cellulitis or contact dermatitis should be suspected of metastasis derived from an internal malignancy, even for very rare sites of metastasis.
Adenocarcinoma* ; Aged ; Cellulitis* ; Cytoplasm ; Dermatitis, Contact ; Eyelids ; Humans ; Immunophenotyping ; Keratin-20 ; Mucin-1 ; Neoplasm Metastasis ; Orbital Cellulitis ; Skin ; Ulcer

Ovarian endometrioid adenocarcinoma (EAC) with a yolk sac tumor (YST) component is extremely rare. Only twelve cases have been reported in the English literature. We report here two additional cases of this rare tumor. The YST component showed classic microscopic features, and immunohistochemically stained positive for alpha-fetoprotein (AFP), but negative for cytokeratin 7 (CK7), epithelial membrane antigen (EMA), estrogen receptor (ER) and progesterone receptor (PR). The EAC appeared to blend into the YST in several areas and immunohistochemically stained positive for CK7, EMA, ER, and PR, but negative for AFP.
alpha-Fetoproteins ; Aminocaproic Acids ; Carcinoma, Endometrioid ; Endodermal Sinus Tumor ; Estrogens ; Female ; Keratin-7 ; Mucin-1 ; Ovary ; Receptors, Progesterone ; Yolk Sac

Whereas extramammary Paget's disease commonly occurs in the apocrine gland rich skin areas, ectopic extramammary Paget's disease develops in the skin areas that are devoid of apocrine glands. We experienced the case of a 34 year-old female patient who had a skin lesion in the upper outer quadrant of the right breast for 5 years and that lesion was diagnosed as Paget's disease according to the punch biopsy. There was no other underlying malignancy, and so wide excision was performed. The final pathologic diagnosis was Paget's disease confined to the epidermis and the size of the tumor was 3.0x1.1 cm. Positive staining for cytokeratin 7, epithelial membrane antigen and negative staining for S-100 protein and HMB-45 was observed on the immunohistochemical tests. We report here on an extremely unusual case of ectopic extramammary Paget's disease of the breast skin, and we include a review of the relevant literature.
Apocrine Glands ; Biopsy ; Breast ; Epidermis ; Female ; Humans ; Keratin-7 ; Mucin-1 ; Negative Staining ; Paget Disease, Extramammary ; S100 Proteins ; Skin

Combined hepatocellular and cholangiocarcinoma (cHCC-CC) is an uncommon form of primary liver cancer having features of both hepatocellular and billiary epithelial differentiation. We reptitis B virus. A 36-year-old woman was diagnosed by ultrasonography with an asymptomatic tumor inort a case of cHCC-CC that separately developed in a patient who was serologically positive for hepa the left lobe of the liver. Based on radiologic and serologic findings of elevated serum alpha-fetoprotein (AFP) level, a preoperative diagnosis of hepatocellular carcinoma was made. A left lobectomy of the liver was done. On histological examination, the resected tumor was shown to consist of two separated masses. One was hepatocellular carcinoma that showed immunoreactivity for AFP. The other was cholangio- carcinoma that showed immunoreactivity for cytokeratin-19 (CK-19) in a separate form.
Adult ; alpha-Fetoproteins ; Carcinoma, Hepatocellular ; Cholangiocarcinoma* ; Diagnosis ; Female ; Herpesvirus 1, Cercopithecine ; Humans ; Keratin-19 ; Liver ; Liver Neoplasms ; Ultrasonography