BACKGROUND: Seborrheic keratosis(SK) is a benign epidermal tumor histopathologically composed of basaloid cells and squamous cells. OBJECTIVE: We investigated the mode of expression of various epidermal differentiation markers in SK. METHODS: Twenty two cases of pathologically confirmed SK were collected from the pathologic files. The histological types included acanthotic type (19 cases) and reticulated type (3 cases). Immunohistochemical staining to various cytokeratins(CK), involucrin, loricrin and filaggrin was performed. RESULTS: Two expression patterns of CK 1 were observed. There was diffuse homogeneous expression of CK 1 in squamous cells in 14 cases and in 8 cases heterogeneous expression with immunoreactive cells scattered among nonimmunoreactive cells to CK 1. Immunoreactivities to CK 14 were observed in basaloid cells. CK 6 was expressed in entire epidermis including basal layer in 19 cases. But stronger expression of CK 6 was expressed in squamous cells than in basaloid cells. In 3 cases immunoreactive cells to CK6 were squamous cells in spinous and granular layer. CAM 5.2 was focally expressed in basaloid cells only in one case. Expression of involucrin was observed in squamous cells in upper spinous and granular layer. Loricrin and filaggrin were expressed linearly in only uppermost granular layer or with horny layer. CONCLUSION: CK expressions in SK suggest that differentiation process from basaloid cells to squamous cells is the same as that of from basal cells to sqamous cells in the suprabasal epidermis. Squamous cells of SK are activated keratinocytes which proceed normal keratinization with normal involucrin expression although terminal differentiation is presumptive to be slightly retarded because of decreased expression of loricrin and filaggrin.
Antigens, Differentiation* ; Epidermis ; Keratinocytes ; Keratosis, Seborrheic*

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

To evaluate the maturation and differentiation state of cultured keratinocytes, the author investigated expression of differentiation markers in cultured keratinocytes. The specimens were divided into three experimental groups, 3rd passage keratinocytes cultured in serum free media (3rd SFM group), 6th passage keratinocytes cultured in serum free media (6th SFM group) and 3rd passage keratinocytes cultured in DMEM (DMEM group). CK14, marker of basal layer, expressed in all groups. The expression was localized and condensed in the SFM groups but spreade in the DMEM group. Most of the cells in both SFM groups were positive but a few cells in DMEM group were also positive. CK10, marker of initiation of differentiation, expressed weakly in DMEM group but there was no expression in both SFM groups. Involucrin, marker of terminal differentiation, expressed weakly in DMEM group but there was no expression in both SFM groups. CK16 and 17, markers of fast turnover of keratinocytes, were not expressed in SFM groups. Weak positive reactions were observed in DMEM group. With these results the authors concluded that the keratinocytes from 3rd passage to 6th passage, cultured in serum free media with calcium less than 0.1 mM, had highly homogeneous basal cell characteristics.
Antigens, Differentiation ; Calcium ; Culture Media, Serum-Free ; Humans* ; Keratinocytes* ; Keratins*

BACKGROUND: S100A8 is differentially expressed in various cell types and is associated with a number of malignant disorders. S100A8 may affect tumor biology. However, its role in cutaneous squamous cell carcinoma (SCC) is not well established. OBJECTIVE: This study aims to investigate the relationship between S100A8 and cutaneous SCC development. METHODS: We performed immunohistochemical staining to detect S100A8 expression in facial skin specimens of premalignant actinic keratosis (AK), malignant SCC, and normal tissues. In addition, we utilized postconfluence and high calcium-induced differentiation in a culture system model. Furthermore, we constructed a recombinant adenovirus expressing GFP-tagged S100A8 to investigate the role of S100A8 in SCC cell differentiation. RESULTS: S100A8 was significantly overexpressed in human cutaneous SCC compared to that in normal and AK tissues. S100A8 was gradually upregulated in SCC cells in a post-confluence-induced differentiation model. Overexpression of S100A8 in SCC cells induced by adenoviral transduction led to increased expression levels of differentiation markers, such as loricrin, involucrin, and filaggrin. S100A8 overexpression also increased loricrin and involucrin luciferase activity. CONCLUSION: S100A8 regulates cutaneous SCC differentiation and induces well-differentiated SCC formation in skin.
Adenoviridae ; Antigens, Differentiation ; Biology ; Carcinoma, Squamous Cell* ; Cell Differentiation ; Humans ; Keratosis, Actinic ; Luciferases ; Skin

CD28 family consists of CD28, ICOS, CTLA-4 and PD-1 molecules. The former two are activation receptors and the later two are inhibition receptors. They produce co-stimulatory signals combining with the relevant molecules in B7 family, which plays important role in T cell activation and homeostasis among T subsets. Although the mechanism of signaling by CD28 and CTLA-4 has been well studied, many questions still remain to be answered. Further investigations are required for substantiating the dual-signaling model.
Animals ; Antigens, CD ; Antigens, Differentiation ; immunology ; CD28 Antigens ; immunology ; CTLA-4 Antigen ; Humans ; Immunoglobulin Fc Fragments ; immunology ; Signal Transduction

BACKGROUND: Mesenchymal stem cells (MSCs) have strong self-renewal ability and multiple differentiation potential. Some studies confirmed that spreading shape and area of single MSCs influence cell differentiation, but few studies focused on the effect of the circularity of cell shape on the osteogenic differentiation of MSCs with a confined area during osteogenic process.METHODS: In the present study, MSCs were seeded on a micropatterned island with a spreading area lower than that of a freely spreading area. The patterns had circularities of 1.0 or 0.4, respectively, and areas of 314, 628, or 1256 µm² . After the cells were grown on a micropatterned surface for 1 or 3 days, cell apoptosis and F-actin were stained and analyzed. In addition, the expression of β-catenin and three osteogenic differentiation markers were immunofluorescently stained and analyzed, respectively.RESULTS: Of these MSCs, the ones with star-like shapes and large areas promoted the expression of osteogenic differentiation markers and the survival of cells. The expression of F-actin and its cytosolic distribution or orientation also correlated with the spreading shape and area. When actin polymerization was inhibited by cytochalasin D, the shape-regulated differentiation and apoptosis of MSCs with the confined spreading area were abolished.CONCLUSION: This study demonstrated that a spreading shape of low circularity and a larger spreading area are beneficial to the survival and osteogenic differentiation of individual MSCs, which may be regulated through the cytosolic expression and distribution of F-actin.
Actins ; Antigens, Differentiation ; Apoptosis ; Cell Differentiation ; Cell Shape ; Cytochalasin D ; Cytosol ; Mesenchymal Stromal Cells ; Osteogenesis ; Polymerization ; Polymers

The expression of inducible co-stimulator (ICOS) in peripheral blood T lymphocytes from the patients with systemic lupus erythematosus (SLE) and the role in the pathogenesis of SLE was investigated. By using two-color immunofluorescent staining and flow cytometric assay, the expression levels of ICOS in peripheal blood T lymphocytes from 33 patients with SLE and 16 healthy volunteers were detected. SLE diseases activity index (SLEDAI) of the patients with SLE was used to evaluate the disease activity. The correlation between the ICOS expression and SLEDAI was analyzed among the groups. The results showed that the expression levels of ICOS in T lymphocytes in active SLE group was markedly higher than those in the control and inactive SLE groups (both P< 0.01). There was no significant difference in the expression levels of ICOS between the inactive SLE and the control groups (P>0.05). In active SLE and inactive SLE groups, positive linear correlation was found between the levels of the ICOS expression in T lymphocytes and SLEDAI (r=0. 711, P=0.001; r=0.561, P=0.03). It was suggested that the expression of ICOS in peripheral blood T lymphocytes from the patients with active SLE was up-regulated and and ICOS might be related to the pathogenesis of SLE.
Antigens, Differentiation, T-Lymphocyte/*biosynthesis ; Antigens, Differentiation, T-Lymphocyte/genetics ; Lupus Erythematosus, Systemic/blood ; Lupus Erythematosus, Systemic/etiology ; Lupus Erythematosus, Systemic/*immunology ; T-Lymphocytes/*immunology

OBJECTIVETo investigate the association of cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) gene A/G polymorphism with susceptibility to diabetes mellitus in Han Chinese.

METHODSAn A/G transition at position 49 of exon 1 was analyzed in 31 patients with type 1 diabetes, 31 patients with type 2 diabetes, and 36 controls were analyzed by polymerase chain reaction-restriction fragment length polymorphism analysis.

RESULTSA highly significant increase in the frequency of the G allele was seen in patients with type 1 diabetes compared with controls (66.1 % vs. 34.7%, respectively; P < 0.0005; OR = 3.670) . This reflected an increase in the GG genotype in patients (48.4% vs. 22.2%, respectively; P =0.025; OR =3.281) and a significant decrease in the AA genotype (16.1 % vs. 52.8%, respectively; P = 0.002). The allele frequencies of A and G in patients with type 2 diabetes were not significantly different from controls(A/G, 50.0/50.0% vs. 65.3/34.7%; P = not significant) . The distribution of genotype, however, differed significantly. This difference reflected an increase in the AG genotype in patients (54.8% vs.25.0%, respectively; P=0.012; OR=3.643) and a decrease in the AA genotype (22.6% vs. 52.8%, respectively; P=0.011).

CONCLUSIONSCTLA-4 49 AA is protective from diabetes mellitus, whereas, CTLA-4 49 G allele (both as homozygotes and as heterozygotes ) confers an increased risk of diabetes mellitus.

Abatacept ; Antigens, CD ; Antigens, Differentiation ; genetics ; CTLA-4 Antigen ; China ; ethnology ; Diabetes Mellitus ; genetics ; Humans ; Immunoconjugates ; Polymorphism, Genetic

OBJECTIVETo search a method for showing dust granules in phagocytes.

METHODSThe autopsy samples of 10 cases who died of non-pulmonary diseases(group A), 6 cases with silicosis(group B) and 5 cases of dead fetus(group C) were collected. The samples of lung, spleen and lymphnodes were stained by methods of (1)HE staining; (2)Warthin-Starry staining(W-S staining) and modified W-S staining; (3) CD68 (as a first antibody) immunohistochemical staining. The ultrastructure and chemical component of the dust granules in the pulmonary phagocytes(dust cells) of group A(3 cases) and group B(3 cases) were observed and analyzed by transmission electron microscope(TEM) and analytic electron microscope(AEM).

RESULTSIn group A and B smaller or minute dust granules which could not be shown by HE staining method were clearly shown by W-S staining and modified W-S staining methods. There were no positive granules found in group C. The CD68 marker of the dust cells was positive. The size, shape, density and chemical elements of the dust granules of group A were different from those of group B under TEM and AEM, excluding bacteria and other intracellular contents.

CONCLUSIONThe dust granules in the phagocytes could be shown by W-S staining method, its staining effect for minute dust granules is superior to general HE staining. However, the determination of chemical elements of dusts must depend on AEM.

Antigens, CD ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Dust ; Humans ; Microscopy, Electron ; Phagocytes ; ultrastructure ; Silicosis ; pathology ; Staining and Labeling ; methods

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