BACKGROUND AND OBJECTIVES: The aim of this study is to determine whether the hyperproliferative and hyperkeratotic characters of cholesteatoma are associated with differentiation of keratinocytes in cholesteatoma by examining the localization of marker proteins, such as involucrin, filaggrin, and cytokeratins. MATERIALS AND METHODS: Immunohistochemical study was carried out in 30 cholesteatoma tissues and 10 retroauricular skins to examine the expression of involucrin, filaggrin, cytokeratin 4, 10 and 16. The staining results were graded as negative, weakly positive (<10%), moderately positive (10-70%), and strongly positive (>70%). RESULTS: Involucrin was strongly expressed in upper spinous, granular, and corneal layer of cholesteatoma. Filaggrin was strongly expressed in granular and corneal layer of cholesteatoma. Cytokeratin 4 was expressed in basal layer of retroauricular skin, but occasionally expressed in suprabasal layer of cholesteatoma. Cytokeratin 10 was homogenously expressed in all suprabasal layer of retroauricular skin, whereas pattern of shift to surface layer was showed in cholesteatoma. Cytokeratin 16 was moderately expressed at suprabasal layer in cholesteatoma. CONCLUSIONS: It can be suggested that early differentiation of suprabasal layer may lead to hyperdifferentiation and hyperkeratosis. Different expression of cytokeratins possibly indicates the altered differentiation of cholesteatoma.
Cholesteatoma ; Intermediate Filament Proteins ; Keratin-16 ; Keratin-4 ; Keratinocytes ; Keratins ; Protein Precursors ; Proteins ; Skin

BACKGROUND AND OBJECTIVES: The aim of this study is to determine whether the hyperproliferative and hyperkeratotic characters of cholesteatoma are associated with differentiation of keratinocytes in cholesteatoma by examining the localization of marker proteins, such as involucrin, filaggrin, and cytokeratins. MATERIALS AND METHODS: Immunohistochemical study was carried out in 30 cholesteatoma tissues and 10 retroauricular skins to examine the expression of involucrin, filaggrin, cytokeratin 4, 10 and 16. The staining results were graded as negative, weakly positive (<10%), moderately positive (10-70%), and strongly positive (>70%). RESULTS: Involucrin was strongly expressed in upper spinous, granular, and corneal layer of cholesteatoma. Filaggrin was strongly expressed in granular and corneal layer of cholesteatoma. Cytokeratin 4 was expressed in basal layer of retroauricular skin, but occasionally expressed in suprabasal layer of cholesteatoma. Cytokeratin 10 was homogenously expressed in all suprabasal layer of retroauricular skin, whereas pattern of shift to surface layer was showed in cholesteatoma. Cytokeratin 16 was moderately expressed at suprabasal layer in cholesteatoma. CONCLUSIONS: It can be suggested that early differentiation of suprabasal layer may lead to hyperdifferentiation and hyperkeratosis. Different expression of cytokeratins possibly indicates the altered differentiation of cholesteatoma.
Cholesteatoma ; Intermediate Filament Proteins ; Keratin-16 ; Keratin-4 ; Keratinocytes ; Keratins ; Protein Precursors ; Proteins ; Skin

BACKGROUND: Clear cell acanthoma usually appears as an asymptomatic nodule on the leg. It has an unusual clinical feature in that it is presented as chronic eczema on the areola. The origin of clear cell acanthoma is not yet clear, although many hypotheses have been proposed, including a benign neoplasm or an inflammatory dermatosis. OBJECTIVE: In this study, clear cell acanthoma on the areola showing clinically eczematous features were analysed by immunohistochemical techniques, using antibodies against cytokeratin 16, involucrin and PCNA and compared with psoriasis and squamous cell carcinoma. METHODS: Using the immunohistochemical method with formalin-fixed, paraffin-embedded sections, we analysed the expression of cytokeratin 16, involucrin and PCNA in biopsy specimens of 6 cases of clear cell acanthoma on the areola, 5 cases of psoriasis, and 5 cases of squamous cell carcinoma. RESULTS: The expression of cytokeratin 16 was detected in spinous and granular layers in all cases of clear cell acanthoma and psoriasis and three cases of squamous cell carcinoma. Psoriasis showed slightly higher immunoreactivity than clear cell acanthoma and squamous cell carcinoma, but this difference was not statistically significant (p=0.142). The expression of involucrin was detected in spinous and granular layers in all cases of clear cell acanthoma, psoriasis, and squamous cell carcinoma. The immunoreactivities were similar. The expression of PCNA was detected in basal and spinous layers in two cases of clear cell acanthoma, four cases of psoriasis, and five cases of squamous cell carcinoma. The expression of PCNA was higher in psoriasis and squamous cell carcinoma than in clear cell acanthoma, and this difference was statistically significant (p=0.034, p=0.004). CONCLUSION: Clear cell acanthoma on the areola may result from increased psoriasis-like inflammatory proliferation and accelerated differentiation of keratinocytes.
Acanthoma* ; Antibodies ; Biopsy ; Carcinoma, Squamous Cell ; Eczema ; Keratin-16* ; Keratinocytes ; Keratins* ; Leg ; Proliferating Cell Nuclear Antigen* ; Psoriasis ; Skin Diseases

In benign hyperproliferative epidermal diseases(eq. warts, psoriasis) and squamous carcinoma, some molecular markers of hyperproliferative keratinocyte such as cytokeratin 16 and PCNA were expressed predominantly. However, all healthy epidermis including the meatal epidermis are nonreactive to those molecular markers except some of thick skin. Recently, there are several reports which show unusal proliferative capacity around the annular region of the ear drum. Our study has concentrated on the characteristics of the differentiation in healthy deep meatal epidermis using immunohistochemistry with cytokeratins and PCNA. Our investigation has demonstrated that the deep meatal epidermis around the annular region in contrast to the other region of the meatus exhibited unusal proliferative capacity. This result suggests a pathology link such as invasion mechanism and hyperkeratinization between the cholesteatoma and deep meatal skin.
Carcinoma, Squamous Cell ; Cholesteatoma ; Ear ; Epidermis* ; Humans* ; Immunohistochemistry ; Keratin-16 ; Keratinocytes ; Keratins ; Pathology ; Proliferating Cell Nuclear Antigen ; Skin ; Warts

BACKGROUND AND OBJECTIVES: This study investigated the expression of cytokeratin 8, 18, 19 with low molecular weight, which have been classified as a group of simple epithelium-related marker for advanced squamous cell carcinoma of the larynx. MATERIALS AND METHODS: Detection of cytokeratin expression was performed by immunohistochemical study using antikeratin monoclonal antibodies (CAM5.2, RCK108). Immunohistochemical study was used further to detect the presence of p53 mutation in larynx carcinoma, and PCR was performed to detect the infection of HPV. We then tried to draw relationship among these factors with regard to advanced larynx carcinoma. RESULTS: Cytokeratin 8, 18 (CAM5.2) was detected in 17 cases among the 19 advanced larynx carcinoma, and in 3 cases among the 15 normal larynx. Cytokeratin 19(RCK108) was detected in 18 cases among the advanced larynx carcinoma, and in 11 cases among the 15 normal larynx. HPV DNA was detected in 4 of the 19 cases of larynx carcinoma. With regard to subtypes of HPV, HPV 16 was detected in 2 cases. And p53 was detected in 6 out of the 19 cases of larynx carcinoma. There was no correlation among the cytokeratin expression, the p53 expression, and the HPV infection. CONCLUSION: This results show that cytokeratin 8, 18 (CAM5.2) expression might be a meaningful parameter in malignant change of the larynx, but the prognostic role of the cytokeratin and the role of p53 and HPV in cytokeratin expression in larynx carcinoma was not confirmed.
Antibodies, Monoclonal ; Carcinoma, Squamous Cell ; DNA ; Human papillomavirus 16 ; Humans* ; Keratin-8 ; Keratins* ; Larynx ; Molecular Weight ; Papilloma* ; Polymerase Chain Reaction

BACKGROUND: Although many therapeutic agents have been developed, only a few drugs are known to target multiple pathogenic factors in the treatment of acne. OBJECTIVE: The purpose of this study was to identify a new drug candidate, platycodin D, which is a substance extracted from the root of Platycodon grandiflorum. METHODS: Using western blotting and Cell Counting Kit-8 assay, we studied the effects of platycodin D on SEB-1 sebocytes, fibroblasts, and keratinocytes. We investigated its effects in view of lipogenesis, collagen production, anti-inflammatory activity, and dyskeratinization. RESULTS: In SEB-1 sebocytes, platycodin D showed a sebosuppressive effect by downregulating ERK and insulin- like growth factor-1R/PI3K/Akt/sterol-regulatory element binding protein-1 signaling pathways. In addition, adiponectin, one of the adipokines responsible for sebum production, was decreased in platycodin D-treated SEB-1 sebocytes. In fibroblasts, platycodin D increased collagen production and reduced inflammation by inhibiting nuclear factor kappa B and matrix metalloproteinases. Platycodin D also showed anti-inflammatory effects on keratinocytes. It also suppressed keratin 16 expression induced by lipopolysaccharide. Furthermore, platycodin D showed no cytotoxicity on both SEB-1 sebocytes and fibroblasts. CONCLUSION: Our data demonstrate the clinical feasibility of platycodin D for acne treatment and the prevention of acne scarring by sebosuppressive and anti-inflammatory effects, as well as through an increase in collagen levels.
Acne Vulgaris* ; Adipokines ; Adiponectin ; Blotting, Western ; Cell Count ; Cicatrix* ; Collagen* ; Fibroblasts ; Inflammation* ; Keratin-16 ; Keratinocytes ; Lipogenesis ; Matrix Metalloproteinases ; NF-kappa B ; Platycodon ; Sebum

This study was to investigate the effect of subcutaneous injection of the adipose stem cells (ASCs) with conditioned media (CM) in the treatment of acne vulgaris scar. We used Adult male New Zealand white rabbit ears as an animal model and induced acne formation by Kignman method. Adipose tissue was isolated and harvested from the scapula of rabbits, and ASCs were cultured and expanded until passage 1. There have four groups in our experiment, include phosphate buffered saline (PBS), ASCs with PBS (ASC + PBS), CM, and ASCs with CM (ASC + CM) group. This solution of 0.6 ml injected to subcutaneous in each group. ASC + PBS and ASC + CM groups were containing ASCs of 5.0 × 106 cells/ml. We analyzed the treatment of 4 groups to scar tissue after 2 and 4 weeks by hematoxylin and eosin stain, immunohistochemistry, and RNA expression level of tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), and matrix metalloproteinase-2 (MMP-2). Also, the expression of keratin 16 (K16) was detected by western blot analysis. H&E stain showed that infiltration of inflammation cells was significantly reduced at 2 and 4 weeks, as well as re-epithelialization was improved in the ASC + CM group. The ASC + CM gourp was reduced both expression levels of TNF-α, IL-1α, and MMP-2 and K16 protein level. In conclusion, the ASCs with CM has a significant curative effect on acne vulgaris scar, more to the point, the CM has a key role on treatment. It could be applied to a therapeutic approach to regenerate to treat acne vulgaris scar.
Acne Vulgaris ; Adipose Tissue ; Adult ; Blotting, Western ; Cicatrix ; Culture Media, Conditioned ; Ear ; Eosine Yellowish-(YS) ; Hematoxylin ; Humans ; Immunohistochemistry ; Inflammation ; Injections, Subcutaneous ; Keratin-16 ; Male ; Matrix Metalloproteinase 2 ; Methods ; Models, Animal ; Necrosis ; New Zealand ; Rabbits ; Re-Epithelialization ; RNA ; Scapula ; Stem Cells

Objective: To screen the differentially expressed genes (DEGs) in diabetic foot ulcers (DFUs), and to perform functional analysis and clinical validation of them, intending to lay a theoretical foundation for epigenetic therapy of chronic refractory wounds. Methods: An observational study was conducted. The gene expression profile dataset GSE80178 of DFU patients in Gene Expression Omnibus (GEO) was selected, and the DEG between three normal skin tissue samples and six DFU tissue samples in the dataset was analyzed and screened using the GEO2R tool. For the screened DEG, ClusterProfiler, org.Hs.eg.db, GOplot, and ggplot2 in the R language packages were used for Gene Ontology (GO) enrichment analysis of biological processes, molecular functions, and cellular components, and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, respectively. Protein-protein interaction (PPI) analysis was performed using STRING database to screen key genes in the DEG, and GO enrichment analysis of key genes was performed using Cytohubba plug-in in Cytoscape 3.9.1 software. DFU tissue and normal skin tissue discarded after surgery were collected respectively from 15 DFU patients (7 males and 8 females, aged 55-87 years) and 15 acute wound patients (6 males and 9 females, aged 8-52 years) who were admitted to Xiang'an Hospital of Xiamen University from September 2018 to March 2021. The mRNA and protein expressions of small proline-rich repeat protein 1A (SPRR1A) and late cornified envelope protein 3C (LCE3C) were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction and immunohistochemistry, respectively. Data were statistically analyzed with independent sample t test. Results: Compared with normal skin tissue, 492 statistically differentially expressed DEGs were screened from DFU tissue of DFU patients (corrected P<0.05 or corrected P<0.01), including 363 up-regulated DEGs and 129 down-regulated DEGs. GO terminology analysis showed that DEGs were significantly enriched in the aspects of skin development, keratinocyte (KC) differentiation, keratinization, epidermal development, and epidermal cell differentiation, etc. (corrected P values all <0.01). KEGG pathway analysis showed that DEGs were significantly enriched in the aspects of tumor-associated microRNA, Ras related protein 1 signaling pathway, and pluripotent stem cell regulatory signaling pathway, etc. (corrected P values all <0.01). PPI analysis showed that endophial protein, SPRR1A, SPRR1B, SPRR2B, SPRR2E, SPRR2F, LCE3C, LCE3E, keratin 16 (all down-regulated DEGs), and filoprotein (up-regulated DEG) were key genes of DEGs screened from DFU tissue of DFU patients, which were significantly enriched in GO terms of keratinization, KC differentiation, epidermal cell differentiation, skin development, epidermis development, and peptide cross-linking, etc. (corrected P values all <0.01). The mRNA expressions of SPRR1A and LCE3C in DFU tissue of DFU patients were 0.588±0.082 and 0.659±0.098, respectively, and the protein expressions were 0.22±0.05 and 0.24±0.04, respectively, which were significantly lower than 1.069±0.025 and 1.053±0.044 (with t values of 20.91 and 13.66, respectively, P values all <0.01) and 0.38±0.04 and 0.45±0.05 (with t values of 9.69 and 12.46, respectively, P values all <0.01) in normal skin tissue of acute wound patients. Conclusions: Compared with normal skin tissue, there is DEG profile in DFU tissue of DFU patients, with DEGs being significantly enriched in the aspects of KC differentiation and keratin function. Key DEGs are related to the biological function of KC, and their low expressions in DFU tissue of DFU patients may impede ulcer healing.
Female ; Humans ; Male ; Computational Biology ; Diabetes Mellitus/genetics* ; Diabetic Foot/genetics* ; Gene Expression Profiling ; Keratin-16 ; MicroRNAs/genetics* ; Proline ; RNA, Messenger ; Middle Aged ; Aged ; Aged, 80 and over ; Child ; Adolescent ; Young Adult ; Adult ; Wound Healing/genetics*