1.Comparison of the histological morphology between normal skin and scar tissue.
Shao-wei YANG ; Zhi-jun GENG ; Kui MA ; Xiao-yan SUN ; Xiao-bing FU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2016;36(2):265-269
Skin wound healing is a complex event, and interrupted wound healing process could lead to scar formation. The aim of this study was to examine the morphological changes of scar tissue. Pathological staining (HE staining, Masson's trichrome staining, methenamine silver staining) was used to evaluate the morphological changes of regenerating epidermis in normal skin and scar tissue, and immunofluorescence staining to detect the expression of collagen IV, a component of basement membrane (BM), and the expression of integrinβ4, a receptor for BM laminins. Additionally, the expression of CK14, CK5, and CK10 was measured to evaluate the proliferation and differentiation of keratinocytes in normal skin and scar tissue. The results showed that the structure of the skin was histologically changed in scar tissue. Collagen IV, expressed under the epidermis of normal skin, was reduced distinctly in scar tissue. Integrinβ4, expressed in the basal layer of normal skin, was found absent in the basal layer of scar tissue. Additionally, it was found that keratinocytes in scarring epidermis were more proliferative than in normal skin. These results indicate that during the skin wound healing, altered formation of BM may affect the proliferation of keratinocytes, reepithelial and tissue remodeling, and then result in scar formation. Thus, remodeling BM structure during wound repair may be beneficial for improving healing in cutaneous wounds during clinical practice.
Adolescent
;
Adult
;
Cicatrix
;
metabolism
;
pathology
;
Collagen Type IV
;
metabolism
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Female
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Humans
;
Integrin beta4
;
metabolism
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Keratinocytes
;
cytology
;
metabolism
;
pathology
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Male
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Skin
;
cytology
;
metabolism
;
pathology
2.Expression Patterns of Cytokeratins in Cholesteatomas: Evidence of Increased Migration and Proliferation.
Hyung Jong KIM ; Steven P TINLING ; Richard A CHOLE
Journal of Korean Medical Science 2002;17(3):381-388
Aural cholesteatoma is characterized by invading squamous epithelia with altered growth properties. Cytokeratin (CK) expression is affected in epidermal proliferative diseases and represents the alterations of keratinocyte proliferation, differentiation, and migration. In the present study, the intensity of CK immuno-expression was determined, using densitometry at various sites in experimental cholesteatoma in order to characterize changes of keratinocytes. With cholesteatoma formation, CK4, a marker for non-keratinizing epithelia, increased in the suprabasal layers of the annular external auditory canal (EAC) and at the pars tensa indicating an altered differentiation and migration of keratinocytes. CK5/6, a marker of keratinizing squamous epithelium, increased only at the pars tensa of the tympanic membrane, indicating basal keratinocyte hyperplasia. CK1/10 increased in the suprabasal layer at the annular EAC, and at the peripheral pars tensa, indicating increased terminal differentiation of keratinocytes. CK13/16, markers of differentiation and hyperproliferation, increased in suprabasal layer of the EAC, and at the peripheral pars tensa. However, it decreased in the basal layer of the EAC, indicating hyperproliferation and migration of keratinocytes. The findings of this study support the basal cell hyperplasia hypotheses for the pathogenesis of aural cholesteatoma, with regard to hyperproliferation, migration, and an altered differentiation of keratinocytes.
Animals
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Biological Markers
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Cell Division
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Cell Movement
;
Cholesteatoma, Middle Ear/*metabolism/*pathology
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Densitometry
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Gerbillinae
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Keratinocytes/metabolism/pathology
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Keratins/*biosynthesis
3.TRPV1 and TRPA1 in cutaneous neurogenic and chronic inflammation: pro-inflammatory response induced by their activation and their sensitization.
Olivier GOUIN ; Killian L'HERONDELLE ; Nicolas LEBONVALLET ; Christelle LE GALL-IANOTTO ; Mehdi SAKKA ; Virginie BUHÉ ; Emmanuelle PLÉE-GAUTIER ; Jean-Luc CARRÉ ; Luc LEFEUVRE ; Laurent MISERY ; Raphaele LE GARREC
Protein & Cell 2017;8(9):644-661
Cutaneous neurogenic inflammation (CNI) is inflammation that is induced (or enhanced) in the skin by the release of neuropeptides from sensory nerve endings. Clinical manifestations are mainly sensory and vascular disorders such as pruritus and erythema. Transient receptor potential vanilloid 1 and ankyrin 1 (TRPV1 and TRPA1, respectively) are non-selective cation channels known to specifically participate in pain and CNI. Both TRPV1 and TRPA1 are co-expressed in a large subset of sensory nerves, where they integrate numerous noxious stimuli. It is now clear that the expression of both channels also extends far beyond the sensory nerves in the skin, occuring also in keratinocytes, mast cells, dendritic cells, and endothelial cells. In these non-neuronal cells, TRPV1 and TRPA1 also act as nociceptive sensors and potentiate the inflammatory process. This review discusses the role of TRPV1 and TRPA1 in the modulation of inflammatory genes that leads to or maintains CNI in sensory neurons and non-neuronal skin cells. In addition, this review provides a summary of current research on the intracellular sensitization pathways of both TRP channels by other endogenous inflammatory mediators that promote the self-maintenance of CNI.
Animals
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Chronic Disease
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Dendritic Cells
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metabolism
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pathology
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Dermatitis
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metabolism
;
pathology
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Gene Expression Regulation
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Humans
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Inflammation
;
metabolism
;
pathology
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Keratinocytes
;
metabolism
;
pathology
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Mast Cells
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metabolism
;
pathology
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Sensory Receptor Cells
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metabolism
;
pathology
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TRPA1 Cation Channel
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biosynthesis
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TRPV Cation Channels
;
biosynthesis
4.Changes of caspase-8 and caspase-9 activity during apoptosis of keratinocytes induced by trichloroethylene.
Qi-xing ZHU ; Liang-ping YE ; Li-jie WANG ; Tong SHEN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2009;27(4):193-197
OBJECTIVETo observe the change of caspase-8, caspase-9 activity and apoptosis rates in the process of trichloroethylene-induced damage in keratinocytes, and explore the tentative mechanism of apoptosis.
METHODSHuman keratinocytes were exposed to 0.125, 0.250, 0.500, 1.000 and 2.000 mmol/L trichloroethylene for 4, 8, 12 and 24 h. The inhibitive groups were pretreated with 100 micromol/L Z-LEHD-FMK (a specific inhibitor of caspase-9) for 1 h, and were stimulated with 2.000 mmol/l TCE for 12 h. MTT assay was used to detect the viability of different cells; The activity of caspase were calculated according to spectrophotometry; Change of the apoptotic rates was assessed by flow cytometer (FCM) after double-stained with Annexin V-FITC and propidium iodide (PI).
RESULTS(1) The minimum effective concentration for cell viability reduction was 0.125 mmol/L at 12 h and the shortest time required to produce a change was 4 h at a concentration of 2.000 mmol/L (compared with control group, P < 0.01). Cell viability in all the groups markedly decreased from 12 h to 24 h (P < 0.05). (2) The activity of caspase-8 in the various dosage groups at different times had no statistical difference compared with the control group, P > 0.01. (3) At 8 h, 1.000 and 2.000 mmol/L TCE groups could significantly enhance caspase-9 activity (P < 0.05). The caspase-9 activity in all the groups showed differences and was significantly higher than those of control cells when time was over 12 h (P < 0.05). (4) After exposing to different dosages of TCE for 12 h, the rate of apoptosis rose to (80.43 +/- 4.21)% with the increase of dosage, compared with the control group, (9.40 +/- 2.98)%, which showed a dose-effect relationship. (5) The cells pre-treated with caspase-9 inhibitor resulted in a decrease in the caspase-9 activity and apoptosis rates (compared with 2.000 mmol/L TCE exposed group, P < 0.01). However, there was no statistical significance in comparison with the control group (P > 0.05).
CONCLUSIONCaspase-9 may be an important mediator of apoptosis in keratinocytes induced by trichloroethylene.
Apoptosis ; drug effects ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cells, Cultured ; Humans ; Keratinocytes ; drug effects ; enzymology ; pathology ; Trichloroethylene ; toxicity
5.Advances in pathogenesis of psoriasis.
Journal of Zhejiang University. Medical sciences 2006;35(6):673-677
The pathogenesis of psoriasis recently made great advancement due to the introduction of transgenic mouse model. K14-VEGF transgenic mouse showed many of the cellular and molecular features of psoriasis, including angiogenesis in dermis, altered epidermal proliferation and differentiation. Psoriasis of early onset and severe disease showed significantly increased frequency of the +405CC genotype and the C allele. Transgenic mice with keratinocytes expressing active Stat3 (K5. Stat3C mice) developed a skin phenotype closely resembling psoriasis. Stat 3 may link activated keratinocytes and immunocytes required for development of psoriasis. More recently, a novel mouse model with epidermal specific double-knockout of the c-Jun and JunB genes showed developments of psoriasis-like skin phenotype and arthritic lesions. All these data provided more profound understanding in pathogenesis and therapy of psoriasis.
Animals
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Disease Models, Animal
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Humans
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Keratinocytes
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metabolism
;
pathology
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Mice
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Mice, Knockout
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Mice, Transgenic
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Psoriasis
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etiology
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genetics
;
pathology
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STAT3 Transcription Factor
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genetics
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metabolism
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Vascular Endothelial Growth Factors
;
genetics
;
metabolism
6.Inflammatory and Immunomodulatory Effects of Tripterygium wilfordii Multiglycoside in Mouse Models of Psoriasis Keratinocytes.
Shuo ZHANG ; Hong-Jin LI ; Chun-Mei YANG ; Liu LIU ; Xiao-Ying SUN ; Jiao WANG ; Si-Ting CHEN ; Yi LU ; Man-Qi HU ; Ge YAN ; Ya-Qiong ZHOU ; Xiao MIAO ; Xin LI ; Bin LI
Chinese journal of integrative medicine 2024;30(3):222-229
OBJECTIVE:
To determine the role of Tripterygium wilfordii multiglycoside (TGW) in the treatment of psoriatic dermatitis from a cellular immunological perspective.
METHODS:
Mouse models of psoriatic dermatitis were established by imiquimod (IMQ). Twelve male BALB/c mice were assigned to IMQ or IMQ+TGW groups according to a random number table. Histopathological changes in vivo were assessed by hematoxylin and eosin staining. Ratios of immune cells and cytokines in mice, as well as PAM212 cell proliferation in vitro were assessed by flow cytometry. Pro-inflammatory cytokine expression was determined using reverse transcription quantitative polymerase chain reaction.
RESULTS:
TGW significantly ameliorated the severity of IMQ-induced psoriasis-like mouse skin lesions and restrained the activation of CD45+ cells, neutrophils and T lymphocytes (all P<0.01). Moreover, TGW significantly attenuated keratinocytes (KCs) proliferation and downregulated the mRNA levels of inflammatory cytokines including interleukin (IL)-17A, IL-23, tumor necrosis factor α, and chemokine (C-X-C motif) ligand 1 (P<0.01 or P<0.05). Furthermore, it reduced the number of γ δ T17 cells in skin lesion of mice and draining lymph nodes (P<0.01).
CONCLUSIONS
TGW improved psoriasis-like inflammation by inhibiting KCs proliferation, as well as the associated immune cells and cytokine expression. It inhibited IL-17 secretion from γ δ T cells, which improved the immune-inflammatory microenvironment of psoriasis.
Male
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Animals
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Mice
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Tripterygium
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Psoriasis/drug therapy*
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Keratinocytes
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Skin Diseases/metabolism*
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Cytokines/metabolism*
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Imiquimod/metabolism*
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Dermatitis/pathology*
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Disease Models, Animal
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Mice, Inbred BALB C
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Skin/metabolism*
7.Effects of conditioned medium derived from different keratinocytes on proliferation and collagen synthesis of hypertrophic scar fibroblasts.
Shuzhong GUO ; Linxi ZHANG ; Zhen WANG ; Jianbo LIU
Chinese Journal of Plastic Surgery 2002;18(2):83-85
OBJECTIVETo observe the effects of supernatants of normal skin keratinocytes(NK) and scar keratinocytes(SK) on proliferation and collagen synthesis of hypertrophic scar fibroblasts(HSFB).
METHODSThe supernatant, collected from cultured NK and SK, was added to the cultivated HSFB. The MTT-method, 3H-proline incorporation and radioimmunoassay were employed to measure the cell proliferation, collagen synthesis and secretion.
RESULTSNK supernatant could inhibit HSFB proliferation and increase the collagen synthesis, but inhibit collagen secretion, as compared with the control group. On the contrary, SK supernatant could increase collagen synthesis and secretion, which had little effects on HSFB proliferation.
CONCLUSIONKeratinocytes derived from normal skin and hypertrophic scar show different effects on hypertrophic scar fibroblasts.
Cell Division ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Collagen ; biosynthesis ; Culture Media, Conditioned ; Fibroblasts ; physiology ; Humans ; Keratinocytes ; physiology
8.Effect of arecoline on the differentiation of myofibroblasts of oral mucosa.
Xia LI ; Tian-You LING ; Yi-Jun GAO
Chinese Journal of Stomatology 2007;42(7):423-427
OBJECTIVETo investigate the origin of myofibroblasts in oral submucous fibrosis.
METHODSThe oral keratinocytes and fibroblasts were isolated and cultured. The expression of the alpha-smooth muscle actin in the fibroblasts was examined by immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR).
RESULTSNo difference was found in expression of alpha-smooth muscle actin between the fibroblasts that were directly stimulated by arecoline and the control. The expression of alpha-smooth muscle actin in the keratinocyte and fibroblast-cocultured group was higher than in the control group, and higher in fibroblasts cocultured with keratinocytes preprocessed by arecoline than in fibroblasts cocultured with keratinocytes without preprocessed by arecoline.
CONCLUSIONSThe differentiation of myofibroblasts from fibroblasts in oral submucous fibrosis might be induced by the interaction of arecoline and keratinocyte.
Actins ; metabolism ; Arecoline ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Coculture Techniques ; Fibroblasts ; cytology ; metabolism ; Humans ; Keratinocytes ; cytology ; Mouth Mucosa ; cytology ; Oral Submucous Fibrosis ; metabolism ; pathology
9.Increased Expression of Cathelicidin by Direct Activation of Protease-Activated Receptor 2: Possible Implications on the Pathogenesis of Rosacea.
Ji Young KIM ; Yoon Jee KIM ; Beom Jin LIM ; Hyo Jung SOHN ; Dongyun SHIN ; Sang Ho OH
Yonsei Medical Journal 2014;55(6):1648-1655
PURPOSE: Recent findings of increased cathelicidin protein and its proteolytic fragments in rosacea suggest a pathogenic role for cathelicidin in this disease. The relationship between cathelicidin and protease-activated receptor 2 (PAR-2) is therefore of interest, as PAR-2, expressed principally in keratinocytes, regulates pro-inflammatory cytokine expression in the skin. The purpose of this study was to determine the relationship between expression of PAR-2 and cathelicidin in rosacea and to test the effect of direct PAR-2 activation on cathelicidin expression in keratinocytes. MATERIALS AND METHODS: Samples from 40 patients with clinicopathologic diagnosis of rosacea and facial skin tissue samples from 20 patients with no specific findings or milium without inflammation were retrieved. Intensities of immunohistochemical staining for PAR-2 and cathelicidin were compared between normal and rosacea-affected skin tissues. Additionally, correlations between PAR-2 and cathelicidin staining intensities within rosacea patients were analyzed. In cultured keratinocytes, changes in PAR-2, cathelicidin, and vascular endothelial growth factor (VEGF) mRNA and protein were analyzed after treatment with PAR-2 activating peptide (AP). RESULTS: Cathelicidin expression was significantly higher in rosacea skin tissues than in normal tissues (p<0.001), while PAR-2 expression was not significantly higher in rosacea tissues than in normal skin tissues. A positive correlation between PAR-2 and cathelicidin within rosacea samples was observed (R=0.330, p=0.037). After treatment of PAR-2 AP, both mRNA and protein levels for PAR-2, cathelicidin, and VEGF significantly increased in cultured keratinocytes, compared with PAR-2 control peptide treatment. CONCLUSION: PAR-2 may participate in the pathogenesis of rosacea through activation of cathelicidin LL-37, a mediator of innate immune responses in the skin.
Adult
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Aged
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Antimicrobial Cationic Peptides/*metabolism
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Cytokines/metabolism
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Female
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Humans
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Immunity, Innate
;
Inflammation/metabolism
;
Keratinocytes/*metabolism
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Male
;
Middle Aged
;
Receptor, PAR-2/*metabolism
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Rosacea/*pathology
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Skin/pathology
;
Vascular Endothelial Growth Factor A/*metabolism
10.Apoptosis of psoriatic keratinocytes and the severity of patients' condition.
Jing CHEN ; Jian-yun LU ; Zhao-hui TANG ; Cheng-xin ZUO ; Jin-hua HUANG
Journal of Central South University(Medical Sciences) 2006;31(6):936-939
OBJECTIVE:
To investigate the correlation between the apoptosis of psoriatic keratinocytes (KCs) in the psoriatic lesions and patient's condition.
METHODS:
Thirty patients with psoriasis were included. Activated caspase-3 was examined by colorimetric method in skin biopsy specimens. The apoptotic KCs were detected with terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay in skin biopsy specimens. The severities of 26 patients with psoriasis vulgaris (PV) were assessed using the PASI scoring system and the severities of psoriatic lesions.
RESULTS:
Apoptotic indexes (AI) were higher in the pustular psoriatic lesions than those in the vulgaris psoriatic lesions (P<0.05). In 26 patients with PV, AI and activated caspase-3 in the lesions were significantly higher in the progressive stage than those in the static and the regressive stages (P<0.05). There were no correlations between PASI and AI or the expression of activated caspase-3 in PV lesions (P>0.05). The severities of psoriatic lesions were positively correlated to AI or the expression of activated caspase-3 in PV lesions (P<0.05).
CONCLUSION
The amount of apoptotic KCs may be related to varied types, stages of psoriasis, and severities of psoriatic lesions.
Adolescent
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Adult
;
Aged
;
Apoptosis
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Caspase 3
;
metabolism
;
Child
;
Female
;
Humans
;
Keratinocytes
;
pathology
;
Male
;
Middle Aged
;
Psoriasis
;
diagnosis
;
pathology
;
Severity of Illness Index
;
Young Adult