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Multilineage differentiating stress-enduring (Muse) cells are a subgroup of mesenchymal stem cells (MSCs) that express stage-specific embryonic antigen-3 (SSEA-3) and CD105. It is capable of generating cells representing all three germ layers from a single cell and is non-tumorigenic. Muse cells can efficiently migrate and integrate into damaged tissues that produce sphingosine-1-phosphate (S1P), while Muse cells specifically express S1P receptor 2. The factors secreted by Muse cells play anti-inflammatory, anti-fibrosis, and anti-apoptotic effects, and cooperate to complete the function and structure repair of the damaged group. This review summarizes the basic characteristics of Muse cells and describes their application in skin repair.

To investigate the characteristics of multilineage-differentiating stress-enduring (Muse) cells labeled with chloromethyl dialkylcarbocyanine (CM-Dil) in culture and in skin wounds of rats. Normal human dermal fibroblasts (NHDFs) were obtained from foreskins and were confirmed by immunocytochemistry with vimentin. Muse cells were derived from NHDFs using long-term trypsinization (LTT), were confirmed using immunocytochemistry with antibodies against stage specific embryonic antigen-3 (SSEA-3) and CD105 and were expanded in suspension cultures. The Muse cells were labeled with CM-Dil and were further evaluated with respect to their biological properties using CCK-8 assays and scratch tests. One hundred µl CM-Dil-labeled Muse cells at a concentration of 5 × 10

Human epidermal melanocytes can be induced to form melanocyte spheroids and revert to immature characteristics by long-term trypsinization (LTT). To further explore the biological characteristics of melanocytes after LTT and to study the underlying mechanism. Melanocytes were subjected to long-term (2 h) trypsinization in this study, after which their viability, proliferation and autophagy were characterized. The expression of melanocyte markers [human melanoma black45 (HMB45), tyrosinase (TYR) and Nestin] was detected and relative expression levels of mRNAs encoding TYR, Nestin, c-Kit and microphthalmia-associated transcription factor (MITF) were determined. After LTT, more short spindle-shaped melanocytes appeared and viability assays demonstrated that most melanocytes survived that treatment but had decreased proliferation rates compared to the untreated controls. There was a significant increase in autophagy of melanocytes after LTT and the expression of TYR was decreased compared with untreated control melanocytes. There were no significant differences in the expression of HMB45 or Nestin between the two groups. Compared with untreated melanocytes, levels of message ribonucleic acid (mRNAs) encoding TYR, c-Kit and MITF were decreased after LTT, while Nestin mRNA levels were increased. These results clarified that Long-term treatment with trypsin causes the dedifferentiation of mature epidermal melanocytes in vitro.

The aim of this study is to characterize the molecular properties of multilineage differentiating stress-enduring (Muse) cells compared with dermal fibroblasts (FBs) and to characterize differences in their transcriptomes and open chromatin regions that are involved in cellular plasticity. Assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) and RNA sequencing (RNA-seq) analyses was then performed on FBs and Muse cells. Subsequently, cell type-selective gene regulatory regions were identified by coalition analysis. Expression patterns of transcription factors (TFs) and signaling pathways intermediates were verified using quantitative real-time polymerase chain reaction and Western blot analyses. RNA-seq identified 2355 significantly differentially expressed genes (DEGs) that regulate the transcriptome, including 1222 upregulated and 1133 downregulated DEGs. The general panorama of RNA-seq and ATAC-seq analyses confirmed the differences in TFs and open chromatin regions between FBs and Muse cells. ATAC-seq analysis showed that Muse cells had more reproducible and meaningful peaks than FBs, and the peak signals were concentrated near promoter-transcription start site areas. In genomic regions that can be preferentially accessed in FBs and Muse cells, more than 200 TFs had binding motif sequences. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and coalition analyses identified differences in factors involved in the cell cycle and the protein kinase B (AKT) signaling pathway of FBs and Muse cells. The results of RNA-seq and ATAC-seq analyses clarified the genetic basis of the different biological properties of Muse cells and FBs. These results suggest that the cell cycle transition and the AKT signaling pathway may affect the morphology and biological characteristics of Muse cells.

Dermatologists should be aware that the clinical manifestations of syphilis are very complex and changeable. Unilaterally distributed skin lesions and painless lip ulcers may also be the clinical manifestations of secondary syphilis.

To characterize the tolerance of different types of human epidermal cells to trypsinization in vitro and develop a new method to separate and purify melanocytes according to their tolerance to trypsinization. Epidermal cells were obtained by separating the epidermis from human foreskins. Some of those cells were used for routine culture, and then were subjected to differential trypsin digestion. The remaining epidermal cells were resuspended in a 0.25% trypsin solution and then were neutralized by the addition of bovine serum at different time points. Immunofluorescence staining of HMB45, K15 and vimentin was used to identify melanocytes, keratinocytes and fibroblasts, respectively. We found that Keratinocytes, melanocytes and fibroblasts are primary cells obtained from conventional cultures of human skin. Purified keratinocytes and melanocytes can be obtained by conventional differential trypsin digestion, but fibroblasts in the melanocyte population quickly gain a survival advantage after passage. With longer trypsin digestion times, the number of adherent cells decreased, the time required for cell attachment increased, and the proportion of melanocytes increased. There were no obvious keratinocytes in cell populations obtained after 12 h of trypsinization of epidermal cells, and more short spindle-shaped melanocytes appeared, all of which were HMB45-positive. In conclusion, the tolerance of human epidermal melanocytes to trypsinization in vitro was better than epidermal keratinocytes, and that property can be used to purify melanocytes and was better than traditional differential trypsin digestion. The morphology of cells that survived the long-term trypsin digestion changed and they had good proliferative activity, but seemed to be more immature.

This study was designed to characterize the location, morphology and ultrastructure of telocytes (TCs) in human scalp tissue. After obtaining approval for this study and informed consent from the patient, a scalp specimen was obtained. The distribution and morphology of TCs in human scalp tissue was assessed by immunohistochemical staining of CD34 and CD117/c-KIT, and the ultrastructure of TCs was investigated using transmission electron microscopy (TEM). Immunohistochemical staining of CD34 revealed that TCs were located in the connective tissue of human scalp, and were concentrated around hair follicles (HFs), blood vessels, sweat glands, sebaceous glands and adipose lobules. Immunohistochemical staining of CD117 revealed that TCs were mainly located in the dermis of human scalp, surrounding the HFs and sweat glands. Under TEM, TCs were seen and confirmed by their special morphological features. These cells were spindle-shaped, had small cell bodies and long thin processes, and surrounded stem cell clusters in the bulge region of HFs. These results demonstrate that TCs in human scalp were positive for CD34 and CD117, and their strategic positioning surrounding stem cells suggests their possible involvement in local regeneration, remodeling and homeostasis of the skin.

There have been many studies about the formation, storage, transport and degradation of melanosomes in epidermal melanocytes but studies of melanocytes and melanosomes in fetal hair follicles (HFs) have been limited and ambiguous. The goal of this study was to investigate the distribution of melanocytes and the degradation of melanosomes in fetal HFs. After obtaining approval and informed consent for the study, a scalp specimen from a 5 month gestational age fetus was obtained and was divided into two parts. One part was subjected to immunohistochemical staining with the melanocyte-specific marker HMB-45 and was then observed by light microscopy to detect the distribution of melanocytes in HFs. The other part underwent conventional processing for transmission electron microcopy (TEM). Subsequently, the morphology of melanosomes in HF melanocytes and their degradation in cortical keratinocytes were observed. Immunohistochemically, scattered round melanocytes lacking dendrites were mainly observed along the outer root sheath of the lower part of the HF. A few fusiform or tri-dendritic melanocytes were located at the bottom of the hair bulbs. Significantly melanized melanocytes with multiple dendrites were concentrated in the pigmented area in the center of the hair bulbs, only above the dermal papilla. Analysis by TEM revealed melanocytes containing melanosomes at all stages of development. Autophagosomes containing stage mature IV melanosomes were observed in some melanocytes. Many phagolysosomes containing numerous melanosomes were observed in the cortical keratinocytes. Some phagolysosomes were concentrically surrounded by 3-5 layers of endoplasmic reticulum. Melanosomes that had been degraded or were being degraded in phagolysosomes in keratinocytes had lost their integrity and had become an ill-defined melanosomal dust that were arranged irregularly. Partial melanin particles were released into the cytosol. Melanocytes in different regions of fetal HFs had different morphologies and were at various stages of differentiation. Fetal HF melanocytes contained not only melanosomes at different developmental stages, but autophagosomes were seen occasionally. Melanosomes were degraded into irregular pigment particles in the phagolysosomes of cortical keratinocytes. These results provide important clues to elucidate the mechanism of melanosome biodegradation.

Majocchi's granuloma is an uncommon fungal infection of the dermis and subcutaneous tissue. The most frequently identified cause of Majocchi’s granuloma is anthropophilic Trichophyton rubrum, and it is most commonly located on the anterior aspect of the lower limbs in women. Here, we report a case of Majocchi’s granuloma on the forearm, a site that is rarely involved, in a 62-year-old woman who had been bitten by a dog. Histological examination revealed a dense dermal infiltrate composed of lymphoplasmacytic cells and neutrophils, with hyphae in the dermis. The presence of the fungus, Trichophyton tonsurans, was confirmed by mycological examination and molecular methods. Therefore, histological and mycological examination confirmed the diagnosis of Majocchi’s granuloma. The patient was treated with local moxibustion and itraconazole, 200 mg/day, for 60 days, which facilitated a complete resolution of the lesions.

Objective: In order to generate induced Pluripotent Stem Cells (iPSCs) more efficiently, it is crucial to identify somatic cells that are easily accessible and possibly require fewer factors for conversion into iPSCs. Methods: Human epidermal melanocytes were transduced with lentiviral vectors carrying 3 transcription factors (OCT-4, KLF-4 and c-MYC, 3F) or 4 transcription factors (OCT-4, KLF-4, c-MYC and SOX-2, 4F). Once the clones had formed, assays related to stem cell pluripotency, including alkaline phosphatase staining, DNA methylation levels, expression of stem cell markers and ultrastructure analysis were carried out. The iPSCs obtained were then induced to differentiate into the cells representing the three embryonic layers in vitro. Results: Seven days after the transduction of epidermal melanocytes with 3F or 4F, clones were formed that were positive for alkaline phosphatase staining. Fluorescent staining with antibodies against OCT-4 and SOX-2 was strongly positive, and the cells showed a high nucleus-cytoplasm ratio and active karyokinesis. No melanosomes were found in the cytoplasm by ultrastructural analysis. There were obvious differences in DNA methylation levels between the cloned cells and their parental cells. However, there was not a significant difference between 3F or 4F transfected clonal cells. Meanwhile, the iPSCs successfully differentiated into the three germ layer cells in vitro. Conclusion: Human epidermal melanocytes do not require ectopic SOX-2 expression for conversion into iPSCs, and may serve as an alternative source for deriving patient-specific iPSCs with fewer genetic elements.

Objective To evaluate the clinical efficacy of hematoporphyrin monomethyl ether-based photodynamic (HMME-PDT) therapy for the treatment of port-wine stain (PWS) and its sonographic changes. Methods A total of 45 patients with confirmed PWS were enrolled from the Department of Dermatology, Third Affiliated Hospital of Soochow University from March 2017 to June 2018, including 5 with pink PWS, 39 with purplish red PWS and 1 with thickened PWS. All the patients received 3 sessions of HMME-PDT therapy. The skin thickness and density were compared before and after the treatment by using high-frequency ultrasound. Ranked data were analyzed by using nonparametric test. Measurement data were expressed as mean ± standard deviation, and analyzed using ony-way analysis of variance. Multiple comparisons were performed using Student-Newman-Keuls-q (SNK-q) test. The results were considered to be statistically significant if P < 0.05. Results Among the 45 patients with PWS who completed the treatment and follow-up, 10 were cured, 21 received marked improvement, 12 received improvement, and 1 showed no response. The total response rate was 97.78%, and the response rate in the patients with pink PWS was higher than that in the patients with purplish red PWS (U = 12.50, P < 0.001) . The difference value of the skin thickness or skin density before and after the treatment significantly differed among the cured patients, patients receiving marked improvement and those receiving improvement (skin thickness:0.65 ± 0.21, 0.56 ± 0.88, 0.37 ± 0.12 mm respectively; skin density: -8.65 ± 2.19, -6.86 ± 2.79, -4.92 ± 2.91 g/cm3 respectively; F = 14.528, 5.428 respectively, both P < 0.001) , and the difference values of the skin thickness and density were significantly higher in the cured patients than in those receiving improvement (q = 5.82, 4.63, both P < 0.05) . Erythematous swelling to different extents occurred at the laser-exposed sites in the zygomatic and cheek region in 23 patients with PWS and in the frontal-zygomatic region in 6 with PWS after the HMME-PDT therapy, but gradually regressed about 1 week later. Pale brown crusts were observed at the laser-exposed sites in 35 patients, and shed spontaneously about 3 weeks later. Post-inflammatory hyperpigmentation at the laser-exposed sites was observed in 4 patients, and gradually regressed after 2-month follow-up. Conclusions HMME-PDT therapy is effective for the treatment of PWS, with high safety and few adverse reactions. High-frequency ultrasound can be used for objectively evaluating the clinical efficacy of HMME-PDT therapy. Key words: Port-wine stain; Hematoporphyrin photoradiation; Ultrasonography

Background: Autologous melanocyte transplantation plays an important role in the treatment of vitiligo. Objective: Previous studies have indicated that, compared with melanocytes growing in monolayers, melanocyte spheroids have a better survival in growth factor- and serum-deprived conditions. Methods: Melanocyte spheroids were obtained from human epidermis by repetitive long-term trypsinization and maintained an aggregated morphology for a short period in certain conditions. Results: Melanocyte spheroids were capable of growing into normal dendritic melanocytes in monolayer when they were harvested and reinoculated in 24-well plates. Immunohistochemical analysis of the melanocyte spheroids revealed that they were positive for HMB45, a melanosome-specific marker. No melanomas occurred when melanocyte spheroids were transplanted into mice. Conclusion: Our study provides a promising approach for melanocyte transplantation to treat vitiligo.

Induced pluripotent stem cells (iPSCs) play an important role in cell replacement therapy. Several studies have shown that keratinocytes are promising reprogrammed cells. We easily and efficiently enriched epidermal stem cells by attaching them for a limited time in culture dishes. Individual epidermal cells enriched in stem cells, which showed strong immunostaining for K15, were obtained and generated iPSCs within 10 days after transfection with lentiviruses encoding 4 transcription factors (OCT4, SOX2, KLF4, and NANOG). Immunofluorescent staining showed that those iPSCs expressed SOX2, OCT4, NANOG, and SSEA3 (a specific marker of embryonic stem cells). The embryoid bodies generated from those iPSCs stained positively for OCT4 and NANOG and also with the CDy1 dye that is specific for stem cells. When the iPSCs were subcutaneously injected into 4-week-old BALB/c nude mice, teratoma developed at the inoculation site. The iPSCs also demonstrated reduced DNA methylation compared with the original cells and could be induced to differentiate into adipocytes (mesodermal), hepatocytes (endodermal), and neural cells (ectodermal) in vitro. Our research provides an easy and efficient method for producing iPSCs from keratinocytes, which has important applications in cell replacement therapy.

Background Acrylate/acrylamide copolymers have excellent optical properties and biocompatibility and are ideal biomaterials that have been widely used in tissue engineering. Multilineage-differentiating stress-enduring cells (Muse cells) are a specific subset of mesenchymal stem cells that have an excellent potential for the regenerative medicine. Objective This study was designed to investigate the effects of acrylate/acrylamide copolymers on the adhesion, proliferation and pluripotent-like properties of Muse cells, which were derived from normal human dermal fibroblasts by long-term trypsin incubation. Methods In an initial experiment, Muse cells were seeded on primary microarrays containing micro-spots of 275 different mixtures of acrylate/acrylamide. Each mixture was composed of two of 11 different monomers in various proportions, and was replicated in four micro-spots each. According to the adhesion and growth characteristics of Muse cells on those substrates, specific polymer candidates for Muse cells were selected and secondary microarrays were prepared. We then observed the effects of those specific polymer candidates on the adherence, proliferation and differentiation of Muse cells and suitable candidates for their optimal culture were identified. Results According to the adhesion and growth patterns of Muse cells on the primary microarrays, ten suitable mixtures of acrylate/acrylamide copolymers were identified. Muse cells grew well on six of those combinations and around the four other combinations of those polymer mixtures. Muse cells cultured on three of those combinations proliferated and differentiated into long spindle-shaped cells that looked like fibroblasts, while Muse cells cultured on one combination formed clusters that were ring-shaped. Muse cells cultured on some of those combinations of acrylate/acrylamide proliferated and formed clusters that appeared to be very healthy, whereas Muse cells cultured on other combinations formed clusters that expanded outwards. Conclusions These results identified a polymer combination that was optimum for the adhesion, proliferation and maintenance of Muse cells in an undifferentiated state.

The aim of the present study was to explore the effect of the topical application of calcipotriol on the expression levels of zinc finger protein A20 and nuclear factor-κB (NF-κB) in the skin lesions of patients with psoriasis vulgaris. The calcipotriol ointment was topically applied twice a day for 6 weeks by 26 patients with psoriasis vulgaris. At the end of weeks 2, 4 and 6 after the first application of calcipotriol ointment, the clinical efficacy and Psoriasis Area and Severity Index (PASI) score were compared with those prior to treatment. The expression of zinc finger protein A20 and NF-κB in the skin lesions prior to and following treatment with calcipotriol was measured by immunohistochemical staining and western blotting. At the end of week 6, the clinical effectiveness rate of calcipotriol was higher compared with that at the end of weeks 2 and 4 (χ2=8.12 and 9.06, respectively; P

Melkersson-Rosenthal syndrome, plasma cell cheilitis, and nonvenereal sclerosing lymphangitis of the penis are elaborately chosen in this chapter.

Non-inherited syndromes of dermatological interest are classified together in this chapter: Ascher syndrome, Baboon syndrome, blue rubber bleb nevus syndrome, Cronkhite-Canada syndrome, Favre-Racouchot syndrome associated with eyelid papilloma, Gorham syndrome, vulvovaginal-gingival lichen planus, Laugier-Hunziker syndrome, Parry-Romberg syndrome with subclinical cerebral involvement, and Marshall-white syndrome.

Metabolic and nutritional skin diseases presented in this chapter comprise tuberous xanthoma associated with aortic stenosis, xanthoma disseminatum, lipoid proteinosis, necrobiosis lipoidica, juvenile xanthoma, necrobiotic xanthogranuloma, adult xanthogranuloma, congenital erythropoietic porphyria, poikiloderma-like cutaneous amyloidosis, nodular amyloidosis, lichen myxedematosus, scleromyxedema, follicular mucinosis, reticular erythematous mucinosis, cutaneous focal mucinosis, acral persistent papular mucinosis, pellagra, carotenemia, glycogen storage disease type I, and alkaptonuric ochronosis.

Atrophic subcutaneous fat is the chief complaint in cytophagic histiocytic panniculitis, sclerosing panniculitis, eosinophilic panniculitis, generalized lipodystrophy, and lipodystrophia centrifugalis abdominalis infantilis, as illustrated in this chapter.

The skin is especially vulnerable to many physical triggering factors, causing many distinctive clinical features as shown in this chapter: erythema ab igne, actinic lichen planus, phytophotodermatitis, colloid milium, and dermatitis artefacta.

The authors present the reader with several venereal diseases: multiple penile chancres, nodular secondary syphilis, syphilitic alopecia, and viral flat condyloma.

This chapter consists of pretibial myxedema, calcinosis cutis, idiopathic scrotal calcinosis, unilateral nevoid acanthosis nigricans, malignant acanthosis nigricans complicated by gastric adenocarcinoma, hyperandrogenism, insulin resistance and acanthosis nigricans syndrome, and necrolytic migratory erythema.

Skin tumor is the most intriguing topic for the clinicians. This chapter covers a wide range of issues and provides with informative clues and tips associated with clinical features and pathological presentations and keeps pace with the latest development in molecular biology and immunohistochemistry to assist the readers in the establishment of accurate diagnosis. These following disorders are elaborately selected and arranged from a histopathological perspective.

Defects of dermal elastic fibers are relatively uncommon. This chapter touches upon disseminated elastosis perforans serpiginosa, papular elastorrhexis, congenital cutis laxa, cutis laxa, blepharochalasis, pseudoxanthoma elasticum, cutis hyperelastica complicated by the abnormity in immunoassay, giant connective tissue nevus, eruptive collagenoma, white fibrous papulosis of the neck, and pseudoainhum.

Various cutaneous vascular diseases are presented in this chapter: eruptive recurrent pigmented telangiectasia, telangiectasia macularis eruptiva perstans, unilateral nevoid telangiectasia superimposed on the Bier spots, Mondor’s disease, arteriosclerosis obliterans, and eruptive pseudoangiomatosis.

This chapter presents with granulosis rubra nasi, Fox-Fordyce disease, ophiasis and sisaipho hair regrowth pattern, pseudopelade of Brocq, alopecia areata sparing white hairs within lesion, coexistence of acquired hypertrichosis and scalp alopecia, woolly hair, monilethrix, pili annulati, trichonodosis, hair cast, trichostasis spinulosa, cutaneous pili migrans, red hair, permanent poliosis after plucking, congenital leukonychia totalis, hyperpigmentation of the nail from lead deposition, longitudinal erythronychia, chromonychias, subungual splinter hemorrhage, sebaceous gland hyperplasia, as well as sebaceous hyperplasia within epidermis after scald.