In humans, skin barrier dysfunction is thought to be responsible for enhanced penetration of allergens. Similar to conditions seen in humans, canine atopic dermatitis (CAD) is characterized by derangement of corneocytes and disorganization of intercellular lipids in the stratum corenum (SC) with decreased ceramide levels. This study was designed to evaluate the effects of a moisturizer containing ceramide on dogs with CAD. Dogs (n = 20, 3~8 years old) with mild to moderate clinical signs were recruited and applied a moisturizer containing ceramide for 4 weeks. Transepidermal water loss (TEWL), skin hydration, pruritus index for canine atopic dermatitis (PICAD) scores, and canine atopic dermatitis extent and severity index (CADESI) scores of all dogs were evaluated. Skin samples from five dogs were also examined with transmission electron microscopy (TEM) using ruthenium tetroxide. TEWL, PICAD, and CADESI values decreased (p < 0.05) and skin hydration increased dramatically over time (p < 0.05). Electron micrographs showed that the skin barrier of all five dogs was partially restored (p < 0.05). In conclusion, these results demonstrated that moisturizer containing ceramide was effective for treating skin barrier dysfunction and CAD symptoms.
Animals ; Ceramides/*therapeutic use ; Cholesterol/*therapeutic use ; Dermatitis, Atopic/complications/drug therapy/physiopathology/*veterinary ; Dog Diseases/*drug therapy/etiology/physiopathology ; Dogs ; Emollients/*therapeutic use ; Epidermis/drug effects/physiopathology/ultrastructure ; Fatty Acids, Nonesterified/*therapeutic use ; Female ; Male ; Microscopy, Electron, Transmission/veterinary ; Pruritus/drug therapy/etiology/physiopathology/veterinary ; Republic of Korea ; Ruthenium Compounds/chemistry ; Water Loss, Insensible/drug effects

This study was performed to determine the ability of eliminating sodium nitrite and blocking nitrosamine synthesis by anthocyanin from the skin of Alpinia galanga. purified by macroporous resin. The test was conducted under the condition of the simulated human gastric juice (pH 3.0, 37 degrees C) with VitC as positive control. The results showed that the max capability of eliminating sodium nitrite was 87.14%, which is 1.6 times sronger than that of VitC, and the max capability of blocking nitrosamine synthesis was 97.82%, which is 8 times sronger than that of VitC.
Alpinia ; chemistry ; Anthocyanins ; isolation & purification ; pharmacology ; Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Gastric Juice ; chemistry ; Humans ; Hydrogen-Ion Concentration ; Hydrolysis ; drug effects ; Nitrosamines ; antagonists & inhibitors ; metabolism ; Plant Epidermis ; chemistry ; Sodium Nitrite ; metabolism

OBJECTIVETo study the effect of Badu Shengji San (BDSJS) and its decomposed recipes on morphological changes of injured skin in rats.

METHODSD rats with injured skin were treated with BDSJS and its different decomposed recipes for consecutively 14 days. Morphological changes in the injured skin were observed by H&E staining.

RESULTMercury and lead-containing ingredients significantly decreased epidermal thickness and caused vascular hemorrhage, hyperemia and inflammatory cell infiltration in reticular layer of dermis. The compatible herbs alleviated epidermal thickness and reduced dermal lesions.

CONCLUSIONBDSJS' mercury and lead-containing ingredients can accelerate the healing of skin wound and its compatible herbs can relieve the dermis injury induced by mercury and lead.

Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; toxicity ; Epidermis ; drug effects ; injuries ; pathology ; Hemorrhage ; chemically induced ; Hyperemia ; chemically induced ; Lead ; toxicity ; Male ; Mercury ; toxicity ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Skin ; drug effects ; injuries ; pathology ; Skin Diseases ; drug therapy ; pathology ; Wound Healing ; drug effects

OBJECTIVETo evaluate the utility of the cross-species screening strategy for investigating key molecule(s) involved in onset and progression of hepatocellular carcinoma (HCC).

METHODSHCC-related molecule data from our previous studies and in the literature were collected to establish a cross-species dataset. Tissue samples of HCC, non-HCC surrounding liver (para-HCC), and normal liver that were collected from humans, tree shrews and rats. The genes reported to have the most differential expression in HCC were verified by analyzing the mRNA and protein levels by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively.

RESULTSThe cross-species dataset of HCC-related molecules included four genes: epidermal fatty acid-binding protein (E-FABP), liver (L)-FABP, tyrosine a-ketoglutarate transaminase (TKT), and cytokeratin (CK8). In humans, E-FABP mRNA expression was significantly higher (P less than 0.05) in HCC (0.87+/-0.14 vs. para-HCC: 0.64+/-0.12 and normal liver: 0.67+/-0.07; F=20.910). Similar results were obtained in tree shrew (HCC: 0.87 +/- 0.25 vs. para-HCC: 0.73 +/- 0.19 and normal liver: 0.68+/-0.19; F=3.807) and rat (HCC: 0.97+/-0.22 vs. para-HCC: 0.78+/-0.16 and normal liver: 0.80 +/- 0.13; F=4.482). The Western blotting analyses revealed a similar statistically significant trend.

CONCLUSIONThe cross-species screening strategy for tumor genes may represent a feasible and convenient process of identifying key molecule(s) for human HCC. E-FABP may be a particularly crucial molecule for hepatocarcinogenesis.

Adult ; Aged ; Animals ; Carcinoma, Hepatocellular ; metabolism ; Case-Control Studies ; Epidermis ; chemistry ; Fatty Acid-Binding Proteins ; metabolism ; Female ; Humans ; Liver ; metabolism ; Liver Neoplasms ; metabolism ; Male ; Middle Aged ; Rats ; Tupaiidae ; metabolism

BACKGROUNDKeratinocyte growth factor (KGF) significantly influences epithelial wound healing. The aim of this study was to isolate KGF phage model peptides from a phage display 7-mer peptide library to evaluate their effect on promoting epidermal cell proliferation.

METHODSA phage display 7-mer peptide library was screened using monoclonal anti-human KGF antibody as the target. Enzyme linked immunosorbent assay (ELISA) was performed to select monoclonal phages with good binding activity. DNA sequencing was done to find the similarities of model peptides. Three-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, immunofluorescence assay and quantitative real-time PCR analysis were employed to evaluate the effect of the phage model peptides on epidermal cells.

RESULTSThirty-three out of fifty-eight (56.9%) of the isolated monoclonal phages exhibited high binding activity by ELISA. Ten of fifteen obtained phage model peptides were similar to KGF or epidermal growth factor (EGF). MTT assay data showed that four (No. 1 - 4) of the ten phage model peptides could promote epidermal cell proliferation. The expression of keratinocyte growth factor receptor (KGFR) mRNA in the KGF control group and the two phage model peptide groups (No. 1 and No. 2) increased. Expression of c-Fos mRNA and c-Jun mRNA in the KGF control group increased, but did not increase in the four phage model peptide groups (No.1 - 4).

CONCLUSIONFour phage model peptides isolated from the phage display 7-mer peptide library can safely promote epidermal cell proliferation without tumorigenic effect.

Cell Proliferation ; drug effects ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Epidermis ; cytology ; Fibroblast Growth Factor 7 ; chemistry ; pharmacology ; Humans ; Peptide Library ; Peptides ; chemistry ; pharmacology ; Polymerase Chain Reaction ; Receptor, Fibroblast Growth Factor, Type 2 ; genetics

OBJECTIVETo accumulate taxonomic data of the leaf epidermal features of the medicinal species of Euonymus.

METHODTwenty-nine materials of 21 taxa (including 16 species, 4 varieties and 1 form) representing 5 sections of Euonymus are examined by using both of light microscopy and scanning electron microscopy.

RESULTThe form of epidermal cells in Euonymus is usually polygonal or irregular. The stomata were anomocytic in all the species examined except E. maackii and E. bungeanus var. semipersistens. Stomatal types of all species studied may be anomocytic, anisocytic, cycolocytic or the transitional types among them.

CONCLUSIONThe results show that some characteristics (including cuticular membrane, shape of guard cells, inner margin of outer stomatal rim, outer stomatal rim and stomata type) of the leaf epidermis can provide some anatomical evidence for the classification. The characteristics of leaf epidermis support following treatments: E. acanthocarpus var. longipes, E. acanthocarpus var. scandens and E. acanthocarpus var. sutchuanensis should be merged into E. acanthocarpus; E. bungeanus var. semipersistens should be merged into E. maackii; E. hamiltonianus f. lanceifolius should be merged into E. hamiltonianus.

China ; Euonymus ; chemistry ; classification ; Microscopy ; Microscopy, Electron, Scanning ; Plant Epidermis ; chemistry ; classification ; Plant Leaves ; chemistry ; classification ; Plant Stomata ; chemistry ; classification ; Plants, Medicinal ; chemistry ; classification

Ceramides are the main lipid component maintaining the lamellae structure of stratum corneum, as well as lipid second messengers for the regulation of cellular proliferation and/or apoptosis. In our previous study, psoriatic skin lesions showed marked decreased levels of ceramides and signaling molecules, specially protein kinase C-alpha (PKC-alpha) and c-jun N-terminal kinase (JNK) in proportion to the psoriasis area and severity index (PASI) scores, which suggested that the depletion of ceramide is responsible for epidermal hyperproliferation of psoriasis via downregulation of proapoptotic signal cascade such as PKC-alpha and JNK. In this study, we investigated the protein expression of serine palmitoyltransferase (SPT) and ceramidase, two major ceramide metabolizing enzymes, in both psoriatic epidermis and non-lesional epidermis. The expression of SPT, the ceramide generating enzyme in the de novo synthesis in psoriatic epidermis, was significantly less than that of the non-lesional epidermis, which was inversely correlated with PASI score. However, the expression of ceramidase, the degradative enzyme of ceramides, showed no significant difference between the lesional epidermis and the non-lesional epidermis of psoriatic patients. This might suggest that decreased expression of SPT protein is one of the important causative factors for decreased ceramide levels in psoriasis.
Adolescent ; Adult ; Amidohydrolases/*biosynthesis/metabolism ; Apoptosis ; Cell Proliferation ; Ceramidases ; Ceramides/chemistry ; Child ; Epidermis/metabolism ; Female ; Humans ; JNK Mitogen-Activated Protein Kinases/metabolism ; Male ; Models, Biological ; Protein Kinase C-alpha/metabolism ; Psoriasis/*blood/diagnosis ; Serine C-Palmitoyltransferase/*biosynthesis

OBJECTIVETo explore the in vitro methods of isolation and culture of human fetal epidermal stem cells (HFESCs) and the feasibility of the cultured cells as the target cells for gene transfection.

METHODSThe HFESCs were isolated by means of type IV collagen rapid adhering method. The culture medium for HFESCs was prepared according to that for human fetal fibroblasts. The cultured cells were identified by immunohistochemistry staining of keratin-19 and integrin-beta1, cell cycle analysis and clone forming rate determination. Then the cultured cells were gene transfected in vitro by liposome mediating method in which eukaryon expression vector pcDNA3.1/VEGF165 containing vascular endothelial growth factor 165 (VEGF165) were transfected into cultured cells, or by virus vector mediating method in which recombinant adenovirus accompanied vector (raav) containing green fluorescent protein (GFP) (raav/GFP) were transfected into the cultured cells, respectively. The results of in vitro gene transfection of HFESCs were observed by immunohistochemisty staining and fluorescence microscope.

RESULTSHFESCs grew well and formed large clones with higher cloning efficiency and higher ratio of G1 cells than keratinocytes. The cultured cells were strongly positive with immunohistochemistry staining of keratin-19 and integrin-beta1. After being gene-transfected by pcDNA3.1/VEGF165, the VEGF165 of HFESCs showed positive immunohistochemistry staining property, while the HFESCs transfected by raav/GFP exhibited strong fluorescence.

CONCLUSIONHFESCs could be isolated and cultured in vitro by means of rapid adherence to type IV collagen. It seemed feasible that HFESCs were gene transfected with liposome or adeno-associated virus as the vector.

Cell Adhesion ; Cell Cycle ; physiology ; Cells, Cultured ; Endothelial Growth Factors ; genetics ; metabolism ; Epidermis ; Fetus ; G1 Phase ; Green Fluorescent Proteins ; Humans ; Immunohistochemistry ; Integrin beta1 ; analysis ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Keratinocytes ; cytology ; Keratins ; analysis ; Luminescent Proteins ; genetics ; metabolism ; Lymphokines ; genetics ; metabolism ; Microscopy, Fluorescence ; Plasmids ; genetics ; Stem Cells ; chemistry ; cytology ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVETo investigate the different location and the expression characteristics of epidermal stem cells in normal adult skin and scar tissue.

METHODSSkin 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.

RESULTSThe 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.

CONCLUSIONThe 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.

Adult ; Burns ; complications ; metabolism ; pathology ; Cicatrix ; etiology ; metabolism ; pathology ; Epidermis ; chemistry ; cytology ; Humans ; Immunohistochemistry ; Integrin beta1 ; analysis ; Keratin-10 ; Keratin-14 ; Keratins ; analysis ; Middle Aged ; Skin ; chemistry ; cytology ; Stem Cells ; chemistry ; cytology

OBJECTIVETo isolate and culture the murine fetal epidermal stem cells (ESCs) and folliculus pili cells (FPCs) in vitro, and to observe the regeneration of hair follicle and epidermis after cografting of ESCs and FPCs.

METHODSThe ESCs were isolated by adhering to murine type IV collagen and were cultured in conditional medium. The expression level of beta1-integrin and keratin 15 in ESCs was detected. At the same time, the cell cycle and clony forming eficiency (CFE) in ESCs were also determined. The FPCs were isolated and cultured and inoculated in fibrin-gel to form FPCs-gel. A full skin equivalent was prepared by combining ESCs with FPCs-gel and was grafted onto total skin loss wounds on the back of BALB/C nude mice. The histological changes of the wounds and the hair follicles were observed at 8 - 10 weeks after the grafting.

RESULTSThere were high level expressions of beta1-integrin and keratin 15 in murine fetal ESCs. It was indicated by cell cycle analysis that cells in G1 stage accounted for 94.9% of the cells, while that in S stage, 3.5%, suggesting slow cell cycle. Nevertheless, the keratinocytes in G1 stage accounted for 74.1% and that in S stage, 17.5% of cells in control group. The CFE of ESCs was 15.3%, and it was much higher than that in control group (6.7%). The newly formed hair follicles could be found in the grafted rats but not in the control group 8 - 10 weeks after the wound healing in nude rats.

CONCLUSIONThe ESCs could be successfully isolated and cultured in vitro and might participate in the formation of hair follicle structure under the induction of FPCs.

Animals ; Cells, Cultured ; Dermatologic Surgical Procedures ; Epidermis ; Fetus ; Hair Follicle ; cytology ; physiology ; Immunohistochemistry ; Integrin beta1 ; analysis ; Keratin-15 ; Keratins ; analysis ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Regeneration ; Skin ; injuries ; Skin Transplantation ; methods ; Skin, Artificial ; Stem Cells ; chemistry ; cytology ; Wound Healing