Histopathological Differential Diagnosis of Psoriasis and Seborrheic Dermatitis of the Scalp

BACKGROUND: The differential diagnosis of psoriasis and seborrheic dermatitis can be difficult when both conditions are localized to the scalp without the involvement of other skin sites. OBJECTIVE: We aimed to evaluate the histopathological differences between psoriasis and seborrheic dermatitis on the scalp and identify favorable criteria for their differential diagnosis. METHODS: We evaluated 15 cases of psoriasis and 20 cases of seborrheic dermatitis of the scalp that had been clinicopathologically diagnosed. Skin biopsy sections stained with H&E were examined. Additional immunohistochemistry was performed, including Ki-67, keratin 10, caspase-5, and GLUT-1. RESULTS: On histopathological examination, mounds of parakeratosis with neutrophils, spongiform micropustules of Kogoj, and clubbed and evenly elongated rete ridges were significantly more frequently observed in psoriasis. Follicular plugging, shoulder parakeratosis and prominent lymphocytic exocytosis were significantly more common in seborrheic dermatitis. Moreover, significantly higher mitotic figures were observed in psoriatic lesions than in seborrheic dermatitis. Immunohistochemistry did not show any difference between psoriasis and seborrheic dermatitis. CONCLUSION: Histopathological features favoring psoriasis include mounds of parakeratosis with neutrophils, spongiform micropustules of Kogoj, clubbed and evenly elongated rete ridges, and increased mitotic figures (≥6/high-powered field). Features indicating seborrheic dermatitis are follicular plugging, shoulder parakeratosis and prominent lymphocytic exocytosis. Immunohistochemistry was not helpful in differentiating psoriasis from seborrheic dermatitis.

Expression and significance of C/EBPα and CK10 in nasal inverted papilloma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#The expression of C/EBPα, CK10 in nasal inverted papilloma (NIP) were detected in the study. Further discussed their significance in genesia, development and recurrence of NIP.@*METHOD@#Three groups including nasal cavity mucosae (NM 10 cases), nasal polyp (NP 20 cases) and NIP (30 cases) were selected in the study. Expretion of C/EBPα, CK10 were detected by immunohistochemisty PV-6000 method.@*RESULT@#(1) The different expression of C/EBPα and CK10 in the group of NM, NP and NIP was statistically significant (P < 0.05). (2) The different expression of C/EBPα, CK10 in the group of benign NIP and NIP with atypical hyperplasia was statistically significant (P < 0.05). (3) The different expression of C/EBPα and CK10 in the group of NIP with recurrence and NIP with no recurrence was statistically significant, P < 0.05, respectively. (4) Our result indicate that the relationship of C/EBPα and CK10 (r = 0.578, P < 0.01) was direct correlation. The difference was statistically significant.@*CONCLUSION@#In conclusion, the present results describe C/EBPα, CK10 expression in NIP and their possible implication in the regulation of tumor growth and differentiation. C/EBPα and CK10 production may prove useful in terms of a prognostic marker for the recurrence in nasal inverted papilloma.

Expression of microRNA-203 and P63 in human epidermal stem cells and keratinocytes / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the changes in expression of microRNA-203 and P63 in human epidermal stem cells and KCs, and to investigate their effects and significance in the epidermal proliferation and differentiation.</p><p><b>METHODS</b>(1) Five normal foreskin tissue specimens were collected from 5 patients by circumcision in Department of Urinary Surgery of the First Affiliated Hospital of Nanchang University from March to June in 2013. Then single cell suspension was obtained by separating epidermis with trypsin digestion method. The cells were divided into quick adherent cells and non-quick adherent cells by type IV collagen differential adherent method. The biological characteristics of cells were observed by inverted phase contrast microscope immediately after isolation and on post culture day (PCD) 3. The expression of CD29, keratin 19, keratin 1, and keratin 10 was identified by immunocytochemical staining. The expression of microRNA-203 and mRNA of P63 was determined by real-time fluorescent quantitative RT-PCR. The protein expression of P63 was determined by Western blotting. Data were processed with t test and Pearson correlation analysis.</p><p><b>RESULTS</b>(1) Immediately after isolation, quick adherent cells were small, round, and dispersed uniformly. On PCD 3, the cells adhered firmly, and they grew in clones. Immediately after isolation, non-quick adherent cells appeared in different shapes and sizes, and dispersed unevenly. On PCD 3, the cells adhered precariously and did not show clonal growth. Quick adherent cells showed positive expression of CD29 and keratin 19, while non-quick adherent cells showed positive expression of keratin 1 and keratin 10. Quick adherent cells were identified as epidermal stem cells, and non-quick adherent cells were identified as KCs. (2)The expression level of microRNA-203 in epidermal stem cells (0.74 ± 0.20) was lower than that in KCs (3.66 ± 0.34, t =16.582, P <0.001). The mRNA expression level of P63 in epidermal stem cells (4. 16 ± 0.28) was higher than that in KCs (2.90 ± 0.39, t =5. 850, P =0.001). The protein expression level of P63 in epidermal stem cells (1.42 ± 0.05) was higher than that in KCs (0.73 ± 0.03, t =26.460, P <0. 001). (3) The expression level of microRNA-203 was in significantly negative correlation with the expression levels of mRNA and protein of P63 (with r values respectively - 0. 94 and -0.98 , P values below 0.05).</p><p><b>CONCLUSIONS</b>The expression levels of microRNA-203 and P63 in human epidermal stem cells and KCs were significantly different, which might be related to the different characteristics of proliferation and differentiation of the cells.</p>

Bullous ichthyosiform erythroderma: case presentation

Bullous Ichthyosiform Erythroderma [BIE] is a rare disorder of keratinization [mutation in either keratin 1 or 10]. It typically presents with fragile skin, which gives way to gradual evolution of hyperkeratosis. Flaccid blisters, peeling and superficial erosions at sites of minor trauma or friction are apparent within the first few hours of life. Yellow-brown, waxy, ridged or corrugated scale builds up in skin creases, sometimes forming spiny [Hystrix] outgrowths. Cobblestone-like keratoses occur at other sites such as the dorsal hands and feet and over the trunk. We report an 11-year-old boy with a generalized hyperkeratosis on the neck, trunk, extremities and scalp

Screening of differential proteins binding to Nox1 promoter in A549 cell model of inflammation and oxidative stress / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen the regulatory proteins involved in Nox1 promoter activation in a cell model of inflammation and oxidative stress.</p><p><b>METHODS</b>A cell model of inflammation and oxidative stress was established by stimulating A549 cells with tumor necrosis factor-α (TNF-α). The differential proteins binding to Nox1 promoter were screened by DNA pull-down and the binding proteins were separated by 2D electrophoresis and selected according to the their differential expression levels (with over 1.5-fold changes relative to the control level). The screened proteins were finally identified by MALDI-TOF/TOF-MS.</p><p><b>RESULTS</b>Seven differentially expressed protein spots (all upregulated in the cell model) were obtained, among which GLE1, DDX19A, KRT1 and KRT10 were identified by mass spectrometry.</p><p><b>CONCLUSION</b>GLE1, DDX19A, KRT1 and KRT10 participate in the activation of Nox1 promoter in TNF-α-induced A549 cells, and this result provides new insights into the biological roles of the regulatory proteins of Nox1 promoter in inflammation and oxidative stress.</p>

Histologic morphology and involucrin, filaggrin, and keratin expression in normal canine skin from dogs of different breeds and coat types

The purpose of this study was to measure the thickness of canine epidermis at various anatomical sites according to localization of cornified envelopes (involucrin and filaggrin), keratins (keratin 10, 5), and their mRNA expression. This was done in the skin of five breeds of dogs including seven poodles, six golden retrievers, six Shih Tzus, four pugs, and four Labrador retrievers. Epidermal thickness of the stratum corneum and nucleated epidermal layer was significantly different. The greatest thickness was observed in the digital web area and the thinnest epidermis was in the axilla. Epidermal thickness was also significantly different between the breeds (p < 0.05). Immunohistochemical staining scores revealed significant decreases of involucrin, filaggrin, and keratin 10 in the ventral and weight-bearing sites, and a relative increase of keratin 5 (p < 0.05). q-PCR analysis showed that their the levels of mRNA were positively correlated with expression of the corresponding proteins in skin samples (p < 0.05). The present study is the first to report the relationship between epidermal gene expression and histologic morphology of the skin in normal dogs. Further studies will be essential to fully understand the pathogenesis of skin barrier dysfunctions in canines.

Mutation analysis of KRT10 gene in a patient with bullous congenital ichthyosiform erythroderma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the gene mutation in one sporadic case of bullous congenital ichthyosiform erythroderma (BCIE), and to explore the relationship between the genotype and phenotype.</p><p><b>METHODS</b>DNA was extracted from the blood samples of the patient with BCIE, unaffected members of the pedigree, and 50 unrelated healthy controls. PCR was used to amplify the hot spot fragment of keratin 1 (KRT1) and keratin 10 (KRT10) gene. The PCR products were directly sequenced to detect the mutations.</p><p><b>RESULTS</b>A heterozygous 467G>A mutation was found in the patient, resulting in the substitution of arginine (R) by histidine (H) in codon 156 (R156H) in the 1A domain of the KRT10 protein but not in the healthy individuals from the family and the 50 unrelated individuals.</p><p><b>CONCLUSION</b>The mutation of 467G>A in exon 1 of KRT10 gene identified may play a major role in the pathogenic mechanism of this case of BCIE.</p>

Effects of Calcium on the Epidermis in a Skin Organ Culture / 대한피부과학회지

BACKGROUND: Calcium plays a role in the proliferation and differentiation of keratinocytes. In a normal situation, the calcium concentration forms a gradient across the epidermal layers. Calcium is sparse in the basal layer and spinous layer. Skin organ culture is a useful model for conducting research on various aspects of skin biology. Skin organ culture systems are used for defining factors that affect homeostasis when elucidating the modulatory effects of biologic response modifiers, drugs and physical agents on the skin and also when studying complex aspects of cutaneous biology in normal and diseased skin. OBJECTIVE: In this study, we investigated the effects of extracellular calcium on the epidermis in a skin organ culture. METHODS: We compared the skin organ culture patterns under various culture conditions (calcium 0.1, 0.7, 1.4 and 2.0 mM). RESULTS: H&E staining showed different phenotypes according to the calcium concentration and IHC also showed different phenotyes compared to that of keratin 10, involucrin, filaggrin, loricrin and PCNA. CONCLUSION: As a result, we concluded that the calcium gradient is also an important factor in skin organ culture to maintain the vivo-like environment and the appropriate calcium concentration is 1.4 mM.

The changing pattern of stem cell markers of sweat gland in deep partial-thickness burn wound / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the rules of proliferation of epithelial cells of sweat glands in deep partial-thickness burn wound and its transdifferentiation towards epidermal cells during healing process to explore its mechanisms.</p><p><b>METHODS</b>Twenty-eight patients with limbs and trunk burn hospitalized in the Fourth People's Hospital of Taizhou City of Jiangsu Province and the Second Hospital of Shandong University from January 2004 to December 2007 were enrolled in the study. Tissue samples of deep partial-thickness burn wound (DPBW, n = 37), superficial partial-thickness burn wound (SPBW, n = 21), and normal skin (NS, n = 10) were harvested. Expressions of cytokeratin 10 (CK10), bcl-2, P63, CK14 and CK19 of epithelial cells in glandular secretory portion (GSP) in DPBW, SPBW and NS were detected with immunohistochemical double staining method.</p><p><b>RESULTS</b>In NS, CK19, CK14 and CK10 expressed in medium intensity in GSP epithelial cells, P63 and CK14 weakly expressed in basal myoepithelial cells, while no expression of bcl-2 or P63 was observed in all CK10 positive terminally differentiated cells. In SPBW, no change of the construction of GSP and above-mentioned proteins during healing process was observed. In DPBW, as examined on 7(th) post burn day (PBD), expression of P63 and bcl-2 in GSP epithelial cells was enhanced. In DPBW on 8 - 10 PBD, bcl-2, P63, CK19 and CK14 strongly positive solid island-like epithelial structure was formed by proliferation, migration and squamous epithelization of basal cells. Such structure, along with granulation tissue, migrated towards the superficial layer of wounds. The hyperplasia of squamous epithelium resulted in complete reepithelialization. In DPBW, bcl-2, CK14, CK19 and P63 still strongly expressed in hyper-proliferative epidermal basal and suprabasal layers on 13 - 30 day after healing.</p><p><b>CONCLUSIONS</b>During the natural healing process of DPBW, monolayer epithelium (CK19 and CK10 positive) of GSP slowly develops into stratified squamous epithelium (bcl-2, P63, CK19, and CK14 positive), suggesting that the epithelial-epidermal transdifferentiation of GSP undergoes slow retrodifferentiation process of stem cells and transient amplifying cells, resulting in the imbalance between lagged growth of epithelium and the hyperplasia of granulation tissue, constituting one of the important mechanisms of disturbance in DPBW repair.</p>

[Evaluation of expression and intensity of cytiokeratin-10 and 19 in odontogenic keratocyst [OKC] with immunohistochemical staining and comparing it with dentigerous cyst and dental follicle]

Odontogenic keratocyst [OKC] and dentigerous cyst are two most common developmental odontogenic cysts involving jaws. OKC has a high growth potential in contrast to its minor clinical manifestation. It has also a high degree of recurrence rate. The purpose of this study was to diagnose the OKC and dentigerous cysts and also differentiate them based on their immunohistochemical criteria. To provide the materials for this research, we preliminarely reviewed the records of the patients referred to pathology lab of shahid Beheshti Dental College between March 1999 and March 2003 and 60 paraffin-embedded biopsies [20 OKC, 20 dentigerous cysts and 20 dental follicles] were selected. Then immunhistochemical staining was applied to identify cytokeratin-10 [CK-10] and cytokeratin-19 [CK-19] and their conditions, as well. The expression of CK-10, CK-19 are seen in OKC and dentigerous cyst. CK-10 was found more in OKC, along with CK-19 in dentigerous cyst. The expression of these cytokeratins was more in upper half of the epithelial lining of these cysts. One of the dental follicles expressed CK-19, but not CK-10. CK-10 is more specific for OKC as well as CK-19 for dentigerous cyst. These cytokeratins are usually seen in the upper layers of the cyst lining. If an expression of cytokeratin-19 is found in a dental follicle, it may correspond the cystic transformation of the latter one

Three-dimensional Culture Model of the Conjunctival Epithelium

PURPOSE: To reconstruct a cultured conjunctival equivalent that closely resembles normal conjunctival epithelium in three-dimensional culture systems. METHODS: Human conjunctival epithelial cells were cultured on dead de-epidermized dermis in the air-exposed state. After 2 weeks of culture, the sections were stained with hematoxylin and eosin. Immunohistochemical and electron microscopic studies were performed. The results were compared with those of normal conjunctiva and cultured eyelid skin equivalent. RESULTS: In the cultured conjunctival equivalent, nonkeratinizing stratified epithelium was formed similarly to normal conjunctival epithelium. Keratin 13 was expressed, but not keratin 10, in the cultured conjunctival equivalent, similarly to normal conjunctival epithelium. However, in the cultured eyelid skin equivalent, keratinizing stratified epithelium was formed. In addition, keratin 10 was expressed, but not keratin 13, contrary to those of the cultured conjunctival equivalent. In the cultured conjunctival equivalent, ultrastructurally, keratin intermediate filaments and desmosomes were found. In addition, microvilli were seen in the uppermost epithelial cells. CONCLUSIONS: These findings demonstrate that the cultured conjunctival equivalent resembles normal conjunctival epithelium morphologically, biochemically and ultrastructurally, thereby suggesting that the cultured conjunctival equivalent may have a great potential in the study of conjunctival epithelium.

A preliminary study on the identification and distribution of epidermal stem cells in different degrees of burn wounds in scalded rats / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the distribution of epidermal stem cells (ESCs) in different degrees of burn wounds in scalded rats.</p><p><b>METHODS</b>Thirty-two Sprague-Dawley (SD) rats were employed in the study. First degree (I), shallow (shallow II) and deep partial thickness (deep II) and full thickness burn wounds (III) were created on the rat skin. Burn wound samples were harvested at 24 postburn hours (PBHs) from all the wounds and were processed to tissue slices. The tissue slices were stained by immunohistochemistry technique. The expression and distribution of ESCs in different degrees of burn wounds were observed with integrins alpha 2 beta 1 and keratin 10 (K10) as first antibodies.</p><p><b>RESULTS</b>K10 positive cells were found to distribute in the strata spinosum, granulosum and lucidum in the first degree burn wound (I) with large amounts of integrins alpha 2 beta 1 positive cells in the residual basal layer and skin appendages (hair follicles) in shallow partial thickness burn wound (shallow II degree), and there were less integrins alpha 2 beta 1 positive cells in the remaining skin appendages in deep dermis in deep partial thickness burn wound (deep II degree). Finally, integrins alpha 2 beta 1 positive cells were sparsely found in the III degree burn wound.</p><p><b>CONCLUSION</b>The distribution of ESCs in burn wounds was closely related to the depth of burn wound. The residual ESCs might be the origin of burn wound regeneration and reepithelization.</p>

Study on the location and the expression characteristics of epidermal stem cells in normal adult skin and scar tissue / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the different location and the expression characteristics of epidermal stem cells in normal adult skin and scar tissue.</p><p><b>METHODS</b>Skin tissue specimens were harvested from the corresponding sites from 6 healthy volunteers and from scar tissue of 6 patients 1 year after major deep burn. beta1 integrin and keratin 19 (K19) were employed as the biochemical markers for stem cells and transit amplifying cells identification and keratin 14 (K14) and keratin 10 (K10) as markers for post-mitotic cells and terminally differentiated cells respectively. Integrin and keratin were determined by Elivision two-step immunohistochemistry.</p><p><b>RESULTS</b>The expression of beta1 integrin and the K19 positive cell count in the epithelial basal layers of scar tissue were evidently decreased and weakened than those in normal adult healthy skin. Furthermore, the positive cells expressing K14 in epidermis of scar tissue were only located in 2 - 3 layers of basal epidermis, and their number was much less than that in normal adult skin. Whereas the cells positively expressing K10 were distributed wider in area than that in normal healthy skin. The epidermal stem cells and transit amplifying cells in scar epidermis were much less in number than that in normal skin. The differentiation process of scar epidermal stem cells was different from that of normal skin. And the proportions of post-mitotic cells and terminally differentiated cells were abnormal.</p><p><b>CONCLUSION</b>The results indicated that the self-renewal ability of the scar epidermis was decreased, and the differentiation process of it was in disorder, which may be a reason for the abnormality of structure and function of the epidermis in scar, and a reason for the decreased ability of wound healing of scar tissue.</p>

Effect of 12-O-tetradecanoylphorbol 13-acetate (TPA) on the Expression of Keratin in HaCaT Cells / 대한해부학회지

In human skin, specific keratin markers reflect on normal differentiation and pathologic conditions. This experiment focused on the expressional pattern of keratin 10 (K10: normal differentiation marker), and keratin 8 & 13 (K8 & K13: pathologic differentiation marker) together with their cellular localization after treating HaCaT cells with 12-Otetradecanoylphorbol 13-acetate (TPA). The cells were treated with TPA at 0, 0.1, 1 microgram/ml for 2 hours or 6 hours. Morphologic studies revealed that TPA treatment changed the shape of cells into the fibroblast-like cells with highly folded nuclear membrane and reduced number of the desmosome. The results of indirect immunofluorescent staining and Northern blotting analysis showed that TPA considerably down-regulated the expression of K10, while markedly up-regulating the expression of K8 and K13 both at protein and mRNA levels. Furthermore, by simultaneous staining for keratins and DNA content in flow cytometry, it was found that TPA increased the expression of K8 and K13 dramatically at the S-G2-M phase of the cells. In conclusion, these changes induced by TPA in HaCaT cells may indicate a close relationship between the morphologic change and the altered expression of keratin subfamilies. It also suggests that TPA known as a tumor promotor may directly induce the potentially malignant cells even without the support of tumor initiator.

The Measurement of Double Staining for Senescence Associated-beta-Galactosidase Activity and Keratin 10 or Involucrin in Monolayer and Organotypic Cultured Keratinocytes / 대한해부학회지

This experiment developed the methodology of double staining for senescence associated-beta-galactosidase (SA-beta-gal) activity and keratin 10 (K10) or involucrin. To prove the usefulness of the double staining, the author investigated the relationship between senescence and differentiation in monolayer and organotypic cultured keratinocytes. The results were as follows: K10 and involcrin together with SA-beta-gal were doubly stained in most of monolayer cultured keratinocyte. This fact indicated that the senescence and differentiation had simultaneously occurred in the same keratinocyte. In spite of the advantages to preserving structures, the paraffin specimen was not suitable for double staining because of the limitation of SA-beta-gal reactivity. Although the cryosectioned specimen did not have the morphology as good as the paraffin specimen, it was suitable for double staining due to the goodness of SA-beta-gal reactivity. Double staining well reflected the disturbances of senescence and differentiation which could be caused by deranged organizations of the organotypic cultured skin. The organotypic cultured skin which showed deranged organizations such as stratified basal layer, no typical cell features in each epdermal layer, and wide intercellular spaces had SA-beta-gal activity in epidermis and K10 or involucrin reaction in basal cell. But the skin which showed well arranged organizations resembling in vivo skin had no SA-beta-gal activity and no K10 or involucrin reaction in basal cells. In conclusion, it might be suggested that the double staining for SA-beta-gal activity and K10 or involucrin could be used for detecting the extent of senescence and differentiation in the same cell.