Dorsal Skin Tissue Research Articles

Pediococcus acidilactici intake decreases the clinical severity of atopic dermatitis along with increasing mucin production and improving the gut microbiome in Nc/Nga mice

Atopic dermatitis (AD) is a chronic inflammatory skin disease that is associated with intestinal microflora. Since specific probiotics may have better efficacy for AD, we determined the efficacy of Pediococcus acidilactici SRCM102024 (PA) for treating AD in HaCaT cells and NC/Nga mice and explored the mechanism of action. AD-like pathology was induced in HaCaT cells and the dorsal skin of Nc/Nga mice by local exposure to 2,4-dinitrochlorobenzene (DNCB). In AD-lesion induced mice, PA in low-, medium- and high-dosages (5 × 10E6, 5 × 10E7 and 5 × 10E8 CFU/kg bw, respectively) and dexamethasone (3 mg/kg bw, positive-control) were orally administered for 5 weeks. The clinical AD severity, serum immunoglobulin E (IgE) and TNF-α, gene expressions of interleukin (IL)-4, IL-13, and TNF-α and gut microflora were measured. PA treatment (100–300 CFU/mL) dose-dependently increased cell survival in DNCB-induced HACAT cells. PA reduced the relative mRNA expression of PAR-2, TNF-α, IL-4 and IL-13 in the cells. In dorsal skin of Nc/Nga mice applied with DNCB, PA dose-dependently attenuated erythema, hemorrhage, edema, excoriation, dryness and scratching behavior and PA-H improved the clinical symptoms similar to the positive-control. PA-M and PA-H treatment significantly prevented the disturbance of the dorsal skin tissues and decreased the inflammatory cellular infiltrate of mast cells, compared to the control. PA dose-dependently reduced serum IgE and TNF-α concentrations and the mRNA expression of TNF-α, IL-4, and IL-13 in dorsal skin. In gut microflora, relative counts of Lactobacillales, Butyricicoccus and Ruminococcus were decreased in the AD-control compared to the positive-control and the PA-M and PA-H prevented their decrease. However, the positive-control increased serum AST and ALT activities, indicating liver damage as an adverse effect. In conclusion, oral treatment of PA (human equivalent 1 × 10E9-1 × 10E10) relieved the AD symptoms by dose-dependently preventing over-activation of the immune response. Oral PA intake may be a safe and effective alternative therapy for AD.

NCM 1921, a Mixture of Several Ingredients, Including Fatty Acids and Choline, Attenuates Atopic Dermatitis in 1-Chloro-2,4-Dinitrobenzene-Treated NC/Nga Mice.

Atopic dermatitis (AD) is a chronic inflammatory skin disease in humans. In this study, we evaluated the effects of a mixture (NCM 1921) of omega-3 butter, omega-3 beef tallow oil, omega-3 lard oil, caprylic acid, lauric acid, choline, and Fe on AD in 1-chloro-2,4-dinitrobenzene (DNCB)-treated NC/Nga mice. NCM 1921 significantly ameliorated the macroscopic and microscopic signs and reduced skin thickness and mast cell incorporation in the skin lesions of mice with DNCB-induced AD. Furthermore, it reduced serum immunoglobulin E levels; reduced the number of IgE-producing B cells, peripheral blood mononuclear cells, white blood cells, and differential white blood cells; and increased the number of lymphocytes. NCM 1921 normalized the total cell number in dorsal skin tissue, the axillary lymph node, and spleen following DNCB exposure and reduced the number of CD23+/B220+ cells in the axillary lymph node and CD3+ cells in dorsal skin tissue. Moreover, it reduced the levels of interleukin (IL)-4 and IL-13 but increased the levels of interferon-γ in anti-CD3–stimulated splenocytes. Immunohistofluorescence staining showed that NCM 1921 treatment significantly increased claudin1, filaggrin, and Sirt1 protein expressions in AD skin lesions. These results suggest that NCM 1921 could be a valuable remedy for the treatment of AD.

PEPTIDE EXTRACTS FROM TEMPEH PROTECT AGAINST PHOTOAGING VIA DOWN-REGULATING MATRIX METALLOPROTEINASE-3 GENE EXPRESSION

Photoaging damages skin tissue via activation of matrix metalloproteinases (MMPs) and MMP-3 induces activation of MMP-1 and 9. Tempeh is made from fermented soybean in Indonesia and has been known for its high protein content. This study was aimed to investigate the effectiveness of peptide extracts from tempeh against photoaging at gene level in UVB-treated mice. Peptide was extracted from soybean and black soybean tempeh, and identified for its amino acid profiling. Photoaging model was made by gradient UVB exposure to seven groups of mice for 4 weeks. Mice were supplemented with 100 and 300 mg/kg body weight dosages of peptide extracts from soybean and black soybean tempeh. Dorsal skin tissue was collected for histology staining and quantitative polymerase chain reaction (qPCR). This present study found that both tempeh peptide extracts were rich in glutamic acid and branched-chain amino acids. Histology staining showed that tempeh protected mice skin from photoaging. At 100 and 300 mg/kg, both tempeh peptide extracts significantly attenuated MMP-3gene expression that lead to the decrease level of MMP-1 and MMP-9 gene expression. These results suggest that supplementation of both tempeh peptide extracts may possess anti-photoaging effect via altering MMP-3 gene expression in UVB-treated mice.

Effects of metformin on inflammatory status and phosphorylated Stat3 expression in lesions of a psoriasis-like mouse model

Objective To evaluate the effect of metformin on an imiquimod-induced psoriasis-like mouse model, and to explore its related mechanism. Methods A total of 50 male Balb/c mice were randomly divided into several groups: control group, imiquimod group (IM group) and imiquimod combined with metformin group (IM-Met group) . The IM-Met group were classified into 3 subgroups (100, 150 and 200 g/L IM-Met groups) according to the concentration of metformin. Mice in the control group were treated with topical vaseline cream on the back and intraperitoneal injection of 0.9% sodium chloride physiological solution of 0.3 millilitres every day. Mice in the IM group were treated with topical imiquimod cream on the back and intraperitoneal injection of 0.9% sodium chloride physiological solution of 0.3 millilitres every day. Mice in the 100, 150 or 200 g/L IM-Met groups were treated with topical imiquimod cream on the back and intraperitoneal injection of 100, 150 or 200 g/L metformin of 0.3 millilitres respectively every day. The changes of the erythema area, scales and psoriasis area and severity index (PASI) over time in each group were observed by naked eyes. These mice were sacrificed after 7 days of treatment. Western blot analysis was performed to determine the protein expression of interleukin (IL) -17 and phosphorylated signal transducer and activator of transcription 3 (p-Stat3) in the dorsal skin tissues of these mice, and enzyme-linked immunosorbent assay (ELISA) to detect serum levels of IL-17, IL-6 and tumor necrosis factor-α (TNF-α) in these mice. Comparison among these groups was carried out by using one-way analysis of variance, and means of two groups were compared by using least significant difference (LSD) -t test. Results Compared with the IM group, the 100, 150 and 200 g/L IM-Met groups showed alleviated erythema and scaling severity, and significantly decreased PASI score, protein expression of IL-17 and p-Stat3, and serum levels of inflammation factors such as IL-17, IL-6 and TNF-α (all P 0.05, =0.69, =0.76 respectively) . The serum levels of IL-17, IL-6 and TNF-α in the 150 g/L IM-Met group were 33.388 ± 1.556 ng/L, 210.741 ± 4.440 ng/L and 249.434 ± 8.594 ng/L respectively, which were significantly higher than those in the 200 g/L IM-Met group (26.720 ± 2.156 ng/L, 174.997 ± 9.501 ng/L, 193.034 ± 6.661 ng/L respectively; t=7.93, 10.79 and 16.403 respectively, all P<0.05) , but significantly lower than those in the 100 g/L IM-Met group (44.008 ± 1.722 ng/L, 260.926 ± 7.724 ng/L, 271.409 ± 3.037 ng/L respectively; t=-14.47, -17.81 and -7.62 respectively, all P<0.05) . Conclusion Metformin can alleviate psoriasis-like skin lesions and inflammation status in the imiquimod-induced psoriasis-like mouse model to a certain extent, which may be related to the decrease of p-Stat3 protein expression. Key words: Psoriasis; Metformin; Disease models, animal; Interleukins; Tumor necrosis factor-alpha; Signal transducer and activator of transcription 3

Strain-specific ameliorating effect of Bifidobacterium longum on atopic dermatitis in mice

Abstract Atopic dermatitis (AD) is among the most common allergic diseases worldwide. This study evaluated differences in the abilities of Bifidobacterium longum strains to alleviate 2, 4-dinitrofluorobenzene-induced AD in mice. Of the 12 tested B. longum strains, CCFM1029 significantly relieved AD symptoms. This outcome was accompanied by the inhibition of inflammatory cells infiltration in AD-like skin lesions, decreases in serum IgE, and inflammatory cytokines levels in dorsal skin tissue, increases in IFN-γ and IL-10 levels, and a reduction in the cecal propionic acid level. B. longum CCFM1029 also reduced the proportions of unclassified Bacteroidales S24-7, Adlercreutzia, and Allobaculum spp., and restored the gut microbiota structure such that it resembled the control group structure. In conclusion, the administration of B. longum CCFM1029 helped to ameliorate AD by suppressing polarized Th2 immune responses and regulating the gut microbiota and short-chain fatty acids metabolism.

Effects of dietary Arthrospira platensis supplementation on the growth, pigmentation, and antioxidation in yellow catfish (Pelteobagrus fulvidraco)

Spirulina, Arthrospira platensis, contains high levels of protein and lutein. To evaluate nutritional, pigmentation, and antioxidation effects of A. platensis, a total of 900 juvenile yellow catfish (Pelteobagrus fulvidraco) were divided into 18 tanks (3 tanks/treatment, 50 fish/tank) and fed a diet supplemented with A. platensis for 50 days. A. platensis was used in quantities of 0, 57.6, 115.1, 172.7, 230.3, and 287.9 g kg−1 for experimental diets to replace fishmeal protein at levels of 0 (AP0), 20% (AP20), 40% (AP40), 60% (AP60), 80% (AP80), and 100% (AP100). Results revealed that substituting up to 80% of fishmeal by A. platensis biomass did not have a negative effect on fish growth, feed utilization, or apparent digestibility. However, significantly lower growth rates were observed when 100% of the fishmeal was substituted (P < 0.05). A. platensis replacement groups led to an increase in skin yellowness and dose-dependent enrichments of total lutein in dorsal skin, abdominal skin, and liver tissues (P < 0.05). There were similar enhancements observed in glutathione concentrations and glutathione peroxidase activities in plasma and liver (P < 0.05). Analyses based on lutein concentration between abdominal skin and experimental diets revealed that the optimal substitution amount was 72.03% (207.4 g kg−1 of A. platensis), which could ensure growth and pigmentation in yellow catfish.

Effects and mechanism of action of Huang-Lian-Jie-Du-Tang in atopic dermatitis-like skin dysfunction in vivo and in vitro

Ethnopharmacological relevanceAtopic dermatitis (AD), a disorder prevalent during childhood and adulthood, seriously affects the patient's quality of life. Although Huang-Lian-Jie-Du-Tang (HLJDT) has shown anti-inflammatory effects in previous studies, its effects and mechanism of action underlying AD disorder are still largely unknown. ObjectiveThis study explored the anti-inflammatory and immunomodulatory effects of HLJDT on the AD-like dermal disorder, induced in vitro by lipopolysaccharide (LPS)-triggered inflammation, and in vivo by 2,4-dinitrochlorobenzene (DNCB). Materials and methodsIn vivo HLJDT effects were investigated by determining the severity of dermatitis, which consisted of observing signs of skin lesions, visually and through haematoxylin and eosin (HE) staining, in mouse ears and dorsal skin, measuring serum levels of interleukin (IL)-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, interferon (IFN)-γ, the tumour necrosis factor (TNF)-α, and determining the splenic index, number of splenic CD4+/CD8+ T-lymphocytes, as well as the phosphorylation levels of mitogen-activated protein kinases (including MAPKs-p38, ERK, and JNK), IκB-α, and nuclear factor kappa B (NF-κB) (p65) within dermal lesions. Morphological changes in LPS-induced inflammation were observed under a microscope, and ELISA and qPCR assays were used to measure IL-1α, IL-1β, IL-6, and TNF-α expression levels. The protein expression levels of P-ERK/ERK, P-p38/p38, P-JNK/JNK, P-IKβ-α, and P-p65 were measured through western blotting. Additionally, p65 expression was assessed by immunofluorescence, and LPS binding to RAW264.7 cell membrane was studied with laser confocal microscopy. ResultsHLJDT could remarkably mitigate DNCB-induced AD-like lesion symptoms, alleviating inflammatory mediator infiltration in mouse ears and dorsal skin tissue, down-regulating serum expression levels of IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IFN-γ, and TNF-α, normalising the splenic CD4+/CD8+ T-lymphocyte ratio, and inactivating MAPKs (including p38, ERK, and JNK), IκB-α, and NF-κB (p65) in dorsal skin. Furthermore, HLJDT inhibited LPS-induced differentiation of RAW264.7 cells, as evidenced by the decreased protein and mRNA expression of IL-1α, IL-1β, IL-6, and TNF-α. Additionally, it decreased ERK, p38, JNK, IKβ-α, and p65 phosphorylation levels in the MAPKs/NF-κB pathway, inhibited p65 nuclear translocation, and reduced LPS binding to the RAW264.7 cell membrane. ConclusionsHLJDT significantly improved AD-like symptoms via inhibition of the MAPKs/NF-κB pathway. Therefore, administration of HLJDT might be a potential treatment for AD in the clinical setting.

Hydrangea serrata (Thunb.) Ser. Extract Attenuate UVB-Induced Photoaging through MAPK/AP-1 Inactivation in Human Skin Fibroblasts and Hairless Mice.

Skin photoaging is mainly caused by exposure to ultraviolet (UV) light, which increases expressions of matrix metalloproteinases (MMPs) and destroys collagen fibers, consequently inducing wrinkle formation. Nutritional factors have received scientific attention for use as agents for normal skin functions. The aim of this study was to investigate the effect of hot water extracts from the leaves of Hydrangea serrata (Thunb.) Ser. (WHS) against ultraviolet B (UVB)-induced skin photoaging and to elucidate the underlying molecular mechanisms in human foreskin fibroblasts (Hs68) and HR-1 hairless mice. WHS recovered UVB-reduced cell viability and ameliorated oxidative stress by inhibiting intracellular reactive oxygen species (ROS) generation in Hs68 cells. WHS rescued UVB-induced collagen degradation by suppressing MMP expression, and reduced the mRNA levels of inflammatory cytokines. These anti-photoaging activities of WHS were associated with inhibition of the activator protein 1 (AP-1), signal transduction and activation of transcription 1 (STAT1), and mitogen-activated protein kinase (MAPK) signaling pathways. Oral administration of WHS effectively alleviated dorsal skin from wrinkle formation, epidermal thickening, collagen degradation, and skin dehydration in HR-1 hairless mice exposed to UVB. Notably, WHS suppressed UVB activation of the AP-1 and MAPK signaling pathways in dorsal mouse skin tissues. Taken together, our data indicate that WHS prevents UVB-induced skin damage due to collagen degradation and MMP activation via inactivation of MAPK/AP-1 signaling pathway.

Evaluation of dynamic viscoelastic properties of UV-irradiated dorsal skin tissue of hairless mice

Evaluation of dynamic viscoelastic properties of UV-irradiated dorsal skin tissue of hairless mice

The development of simple flow injection analysis tandem mass spectrometric methods for the cutaneous determination of peptide-modified cationic gemini surfactants used as gene delivery vectors

Diquaternary ammonium gemini surfactants are a class of non-viral gene delivery vectors, primarily studied for their dermal applications. However, their biological fate has rarely been investigated. In this work, we developed simple flow injection analysis tandem mass spectrometric methods, (FIA)-MS/MS, to understand the fate and biodistribution of topically applied gemini surfactant-based therapeutics in an ex-vivo skin model.Three peptide-modified gemini surfactants with varied structures and transfection efficiencies were evaluated. For each compound, two methods were developed to quantify their presence in skin tissue and in phosphate buffered saline (PBS). The methods were developed using single-point calibration mode. Skin penetration was assessed on CD1 mice dorsal skin tissue mounted in a Franz diffusion cell after extraction. Amongst the five evaluated liquid-liquid extraction protocols, the Folch method provides the highest extraction efficiency for all compounds. Weak cationic exchange solid phase extraction was also used to further isolate gemini surfactants from endogenous skin lipids. FIA–MS/MS analysis of the skin revealed that all compounds were detected in the skin with minimal partition into the PBS compartment, which represents circulation. Interestingly, the detected amounts of gemini lipids in the skin were correlated with their transfection efficiencies.

Probiotic Lactobacillus sakei proBio-65 Extract Ameliorates the Severity of Imiquimod Induced Psoriasis-Like Skin Inflammation in a Mouse Model.

This study was designed to evaluate the protective effect of ethanol extract (SEL001) isolated from a potent probiotic strain Lactobacillus sakei proBio-65 on imiquimod (IMQ)-induced psoriasis-like skin inflammation in a mouse model. Histopathological and histomorphometrical changes in the ear and dorsal skin tissues were observed under hematoxylin and eosin stain for general histopathological architectures or Masson’s trichrome stain for collagen fibers. The expression profile of psoriasis-associated specific genes was determined using Real-Time PCR analysis. As a result, topical application of IMQ resulted in a significant increase of mean total and epithelial (epidermis) thicknesses, the number of inflammatory cells infiltrated in the dermis, and the decrease of dermis collagen fiber occupied regions in the ear tissues of IMQ and IMQ plus vaseline treated groups when compared to the intact control group. A significant increase of epithelial thickness and number of inflammatory cells infiltrated in the dermis of dorsal skin tissues were also noticed in IMQ and IMQ plus vaseline treated groups as compared to the intact control group, suggesting classic IMQ-induced hypersensitive psoriasis. IMQ-induced hypersensitive psoriasis related histopathological changes to the ear and dorsal skin tissues were significantly inhibited by the treatment of a standard drug clobetasol and SEL001. Further, mRNA expression analysis indicated a significant increase in gene expression levels of pro-inflammatory cytokines, including IL-19, IL-17A, and IL-23 in IMQ and IMQ plus vaseline treated groups than that of the control. Clobetasol and SEL001 treated groups resulted in a lower gene expression level of IL-19, IL-17A, and IL-23 as compared to IMQ and IMQ plus vaseline treated groups. These results enforce that SEL001 could be a novel treatment for psoriasis and an alternative to other drugs that pose a number of side effects on the skin.

Using Terminal Transferase-mediated dUTP Nick End-labelling (TUNEL) and Caspase 3/7 Assays to Measure Epidermal Cell Death in Frogs with Chytridiomycosis.

Amphibians are experiencing a great loss in biodiversity globally and one of the major causes is the infectious disease chytridiomycosis. This disease is caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd), which infects and disrupts frog epidermis; however, pathological changes have not been explicitly characterized. Apoptosis (programmed cell death) can be used by pathogens to damage host tissue, but can also be a host mechanism of disease resistance for pathogen removal. In this study, we quantify epidermal cell death of infected and uninfected animals using two different assays: terminal transferase-mediated dUTP nick end-labelling (TUNEL), and caspase 3/7. Using ventral, dorsal, and thigh skin tissue in the TUNEL assay, we observe cell death in the epidermal cells in situ of clinically infected animals and compare cell death with uninfected animals using fluorescent microscopy. In order to determine how apoptosis levels in the epidermis change over the course of infection we remove toe-tip samples fortnightly over an 8-week period, and use a caspase 3/7 assay with extracted proteins to quantify activity within the samples. We then correlate caspase 3/7 activity with infection load. The TUNEL assay is useful for localization of cell death in situ, but is expensive and time intensive per sample. The caspase 3/7 assay is efficient for large sample sizes and time course experiments. However, because frog toe tip biopsies are small there is limited extract available for sample standardization via protein quantification methods, such as the Bradford assay. Therefore, we suggest estimating skin surface area through photographic analysis of toe biopsies to avoid consuming extracts during sample standardization.

Histological and Morphological Characterization of Developing Dermal Lymphatic Vessels.

The capacity to visualize the lymphatic vasculature in three-dimensions has revolutionized our understanding of the morphogenetic mechanisms important for constructing the lymphatic vascular network during development. Two complementary approaches are commonly employed to assess the function of genes and signaling pathways important for development of the dermal lymphatic vasculature in the mouse embryo. The first of these is whole-mount immunostaining of embryonic skin to analyze dermal lymphatic vessel network patterning and morphology in two and three dimensions. The second is immunostaining of thin tissue sections to examine lymphatic vessel identity, lumen formation and protein localization within discrete lymphatic endothelial cells in a two-dimensional setting. Here we present detailed protocols for multicolor immunofluorescent immunostaining of embryonic dorsal skin and thin tissue cryosections. Each of these methods generates high-resolution images of the dermal lymphatic vasculature, yielding information integral to in-depth characterization of lymphatic vessel phenotypes in the developing mouse embryo.

The origin of Turkish native chickens, Denizli and Gerze

The Wingless-types/beta-catenin (Wnt/β-catenin) signaling pathway plays an important role in embryonic development and affects the physiological development processes of feather follicles. To investigate the role of Wnt/β-catenin pathway in regulating feather follicles morphogenesis, in ovo injection of CHIR-99021, an activator of the Wnt/β-catenin signaling pathway, was conducted in chick embryo model. Initially, a total of 40 embryos were used to assess feather follicles morphogenesis and the expression of β-catenin (E9–E17). The histological results showed that feather follicle morphogenesis was mainly completed from E9 to E17. β-catenin was involved in the processing of the appearance of dermal cell condensation (E9) and the completion of the feather follicles morphogenesis (E17). Next, a total of 160 fertilized eggs were randomly divided into 8 groups for in ovo injection at E9, including a Normal Saline injected group (CON) and the 500, 1,000, 2,000, 5,000, 10,000, 50,000, and 100,000 ng CHIR-99021 groups. Dorsal skin tissue samples were collected at E17 for investigating feather follicles morphology and expressions of β-catenin and lymphoid enhancerbinding factor-1 (LEF1) at gene and protein levels. The results showed that feather follicle diameter in the injected groups were significantly (P < 0.05) increased with limit dose-independence compared to the CON group. CHIR-99021 significantly (P < 0.05) influenced the mRNA expressions of catenin beta-1 (CTNNB1) and downstream target LEF1. In ovo injection of CHIR-99021 caused that β-catenin and LEF1 were significantly (P < 0.05) increased followed the increased doses as determined by western blotting. The immunochemical results showed that β-catenin was detected in the dermal papilla of feather follicles. Given these results, this study suggests to developmental biology that in ovo injection of CHIR-99021 promoted feather follicles morphogenesis and development via activating Wnt/β-catenin signaling pathway and upregulating downstream target LEF1 during embryonic period in chick embryo model. Moreover, CHIR-99021 may be a strong candidate to promote the animal feather/hair industry, especially as a reference for bird feather production.

Bathing effects of east saline groundwater concentrates on allergic (atopic) dermatitis-like skin lesions induced by 2,4-dinitrochlorobenzene in hairless mice

In the present study, it was evaluated whether east saline groundwater concentration solution (ESGWc) exerted a favorable inhibitory effect on 2,4-dinitrochlorobenzene (DNCB)-induced allergic/atopic-like dermatitis (AD). AD was induced and boosted by sensitization with DNCB via topical application on the dorsal back skins. Mice with DNCB-induced AD were bathed in 100-, 200- and 400-fold diluted ESGWc. After 6 weeks bathing, changes to body weight, clinical skin severity scores, scratching behavior, serum total immunoglobulin (Ig)E levels, submandibular lymph node and spleen weights, splenic cytokine levels, skin cytokine mRNA expressions, antioxidant defense systems and superoxide anion productions were recorded to determine the effects of bathing on the histopathology of dorsal back skin tissues. All DNCB-induced mice demonstrated that the induction of AD through IgE-mediated hypersensitivities, oxidative stresses, activation of MMP and apoptosis of keratinocytes resulted in no significant differences in body weight between the different groups at each time point following initial sensitization. However, markers of DNCB-induced AD were significantly inhibited (P<0.05) in a concentration-dependent manner following bathing in all concentrations of ESGWc. The results obtained in the present study suggest that bathing in ESGWc may have favorable protective effects against DNCB-induced AD due to favorable systemic and local immunomodulatory effects, active cytoprotective anti-apoptotic effects, inhibitory effects of matrix metalloproteinase activity, and anti-inflammatory and antioxidative effects.

Clinical application of dorsal perforator flap of the thumb carrying sensory nerves

Objective To investigate the therapeutic effect of including sensory nerves in the dorsal perforator flap of the thumb to cover skin defects. Methods From September 2010 to June 2014, 20 digits in 20 patients with skin defect of the thumb were treated. The defect area was 1.0 cm×1.8 cm to 2.0 cm×2.2 cm in size. Five cases with a defect less than 1.5 cm in the longitudinal dimension were treated with dorsal sensory nerve perforator V-Y advancement flap. Reverse perforator island flap including the thumb dorsal sensory nerve was transferred to cover distal volar or dorsal skin and soft tissue defect of the thumb in 11 cases. Donor sites were directly closed. For 4 cases of distal volar thumb skin defects the donor site of the dorsal sensory nerve reverse perforator island flap could not be directly sutured. A V-Y perforator flap from the dorsum of the thumb was advanced to cover the donor site wounds. The nerve included in the flap was sutured to the dorsal cutaneous nerve or one of the digital nerves to restore flap sensation. Results All 20 flaps survived completely. The patients were follow-up for 6 to 12 months. The color, texture and contour of the flaps were good. Two-points discrimination was 7 to 10 mm on the flap. The function was assessed as excellent in 16 fingers, good in 3 fingers and fair in 1 finger. No complication occurred in the donor site. Conclusion This flap spares the main digital artery and nerve. It is an improvement to the conventional fascia flap and presents as a new method for the treatment of thumb skin defect. Key words: Thumb; Surgical flaps; Perforator

EFFECTIVENESS OF PART LONG THUMB EXTENSOR TENDON DORSAL ULNAR ARTERY CHIMERIC FLAP FOR REPAIR OF Doyle TYPE Ⅲ MALLET FINGER OF THUMB

To investigate the effectiveness of part long thumb extensor tendon dorsal ulnar artery chimeric flap for repair of Doyle type Ⅲ mallet finger of thumb. Between June 2013 and April 2015, 9 cases of Doyle type Ⅲ mallet finger of thumb were treated, which were caused by planer injury. There were 6 males and 3 females, aged from 15 to 65 years (mean, 36 years). The time from injury to operation was 3-8 hours (mean, 5 hours). All cases had interphalangeal joint dorsal skin and soft tissue defects of the thumb; the skin defects ranged from 2.0 cm×1.5 cm to 2.3 cm×2.3 cm; the extensor tendon defect ranged from 0.5 to 1.5 cm in length (mean, 1.0 cm). The part long thumb extensor tendon dorsal ulnar artery chimeric flap of 3.0 cm×2.5 cm to 3.5 cm×3.0 cm in size was used to reconstruct extensor tendon and wound. The donor site was repaired with nasopharyngeal fossa perforating branches pedicled V-Y relay flap. All flaps survived completely and incisions healed by first intention. All patients were followed up 4-12 months (mean, 6 months). The flaps had good color, texture, and contour. At 6 months after operation, the two-point discrimination of chimeric flap was 10-12 mm (mean, 11 mm), and two-point discrimination of relay flap was 12-14 mm (mean, 13 mm). The interphalangeal joint flexion of thumb was 0-40°, and the thumb opposition function was normal. Part long thumb extensor tendon dorsal ulnar artery chimeric flap can repair the Doyle type Ⅲ mallet finger of thumb, which has no injury to the artery and nerve. At the same time the relay flap can achieve linear healing, so good appearance and function of the thumb can be obtained.

Hominis Placenta facilitates hair re-growth by upregulating cellular proliferation and expression of fibroblast growth factor-7.

BackgroundHominis Placenta (HP) known as a restorative medicine in Traditional Chinese Medicine (TCM), has been widely applied in the clinics of Korea and China as an anti-aging agent to enhance the regeneration of tissue. This study was conducted to investigate whether topical treatment of HP promotes hair regrowth in the animal model.MethodsThe dorsal hairs of 8-week-old C57BL/6 mice were depilated to synchronize hair follicles to the anagen phase. HP was applied topically once a day for 15 days. Hair growth was evaluated visually and microscopically. The incorporation of bromodeoxyuridine (BrdU) and expression of proliferating cell nuclear antigen (PCNA), fibroblast growth factor-7 (FGF-7) in dorsal skin tissue was examined by immunohistochemical analysis. Reverse transcription polymerase chain reaction (RT-PCR) was used to measure the mRNA expression of FGF-7.ResultsHP exhibited potent hair growth-promoting activity in C57BL/6 mice. Gross examination indicated that HP markedly increased hair regrowth as well as hair density and diameter. Histologic analysis showed that HP treatment enhanced the anagen induction of hair follicles.Immunohistochemical analysis revealed that BrdU incorporation and the expressions of PCNA were increased by treatment of HP. HP treatment significantly increased the expression of FGF-7, which plays pivotal roles to maintain anagen phase both protein and mRNA levels.ConclusionsTaken together, our results indicate that HP has a potent hair growth-promoting activity; therefore, it may be a good candidate for the treatment of alopecia.

Cornu cervi pantotrichum Pharmacopuncture Solution Facilitate Hair Growth in C57BL/6 Mice.

Objectives:Cornu cervi pantotrichum (CCP) has been widely used in Korean and China, as an anti-fatigue, anti-aging, and tonic agent to enhance the functions of the reproductive and the immune systems. Because CCP has various growth factors that play important roles in the development of hair follicles, we examined whether CCP pharmacopuncture solution (CCPPS) was capable of promoting hair growth in an animal model.Methods:One day after hair depilation, CCPPS were topically applied to the dorsal skin of C57BL/6 mice once a day for 15 days. Hair growth activity was evaluated by using macro- and microscopic observations. Dorsal skin tissues were stained with hematoxylin and eosin. Expressions of bromodeoxyuridine (BrdU), proliferating cell nuclear antigen (PCNA), and fibroblast growth factor (FGF)-7 were examined by using immunohistochemical staining. A reverse transcription polymerase chain reaction (RT-PCR) analysis was also conducted to measure the messenger RNA (mRNA) expression of FGF-7.Results:CCPPS induced more active hair growth than normal saline. Histologic analysis showed enlargement of the dermal papilla, elongation of the hair shaft, and expansion of hair thickness in CCPPS treated mice, indicating that CCPPS effectively induced the development of anagen. CCPPS treatment markedly increased the expressions of BrdU and PCNA in the hair follicles of C57BL/6 mice. In addition, CCPPS up regulated the expression of FGF-7, which plays an important role in the development of hair follicles.Conclusion:These results reveal that CCPPS facilitates hair re-growth by proliferation of hair follicular cells and up-regulation of FGF-7 and suggest that CCPPS can potentially be applied as an alternative treatment for patients with alopecia.

The arresting phase determines the total healing time of a locally irradiated skin wound in swine

ObjectiveRadiation is an important cause of delayed wound healing, and there still exist many questions regarding the patterns and mechanisms of wound healing. This study investigated the characteristics of wound healing after varying doses of local radiation and explored possible causes of the delay in healing caused by radiation. Methods:A full-thickness dorsal longitudinal skin tissue, 2cm in diameter, was excised after local irradiation on one side of the back of swine, and the other side was wounded as a control. The size of the wound area was recorded every two days after injury. Pathological changes, proliferating cell nuclear antigen (PCNA, immunohisto-chemistry) and apoptosis levels (TUNEL assay) were measured at different time points after wounding. ResultsThe course of wound healing can be divided into four phases, namely: the arresting phase, the healing priming phase, the fast healing phase, and the healed phase. Although the total wound healing time was closely correlated to the dose of irradiation (R2= 0.9758), it was more dependent on the length of the arresting phase (R2=0.9903) because once the arresting phase ended, the wound healed at a similar speed regardless of radiation doses. Pathological analysis showed that compared with the control side there were more necrotic tissues, slower epithelial crawling, as well as fewer blood vessels and cellular components in the irradiated side at the arresting phase, while other phases revealed no significant difference concerning these measurements. Immunohistochemistry showed that the irradiated wounds had significantly less PCNA-positive and more TUNEL-positive labeling of cells in the arresting phase than in other phases. Moreover, the changes were positively related to the radiation doses, but there was no obvious difference in cell proliferation or apoptosis among the healing priming phase, fast healing phase or healed phase, whether on the control side or on the irradiated side. ConclusionsAfter local irradiation, the length of the arresting phase determines the wound healing time. Increased apoptosis and decreased cell proliferation might be an important reason for the formation of the arresting phase.