Novel IKZF3 transcriptomic signature correlates with positive outcomes of skin cutaneous melanoma: A pan-cancer analysis.

To investigate the potential relationship between Ikaros family genes and skin cutaneous melanoma (SKCM), we undertook a pan-cancer analysis of the transcriptional signature and clinical data of melanoma through multiple databases. First, 10,327 transcriptomic samples from different cancers were included to determine the overall characteristics and clinical prognoses associated with Ikaros gene expression across cancer types. Second, differentially expressed genes analysis, prognostic evaluation, and gene set enrichment analysis were employed to investigate the role of Ikaros (IKZF) genes in SKCM. Third, we evaluated the relationship between Ikaros family genes and SKCM immune infiltrates and verified the findings using the GEO single-cell sequencing dataset. The results show that Ikaros genes were widely expressed among different cancer types with independently similar patterns as follows: 1. IKZF1 and IKZF3, and 2. IKZF2 and IKZF4-5. IKZF2 and IKZF5 were downregulated in the primary tumor, and IKZF1-3 expression decreased significantly as the T-stage or metastasis increased in SKCM. Moreover, high IKZF1-3 expression was associated with better overall survival, disease-specific survival, and progression-free interval. IKZF3 is an independent prognostic factor of SKCM. Among Ikaros genes, the expression of IKZF1 and IKZF3 positively correlated with the infiltration level of CD4+ T cells and CD8+ T cells, B cells, and Tregs in SKCM and negatively correlated with the infiltration level of M0 and M1 macrophages. Moreover, single-cell sequencing data analysis revealed that IKZF1 and IKZF3 were mainly expressed by immune cells. Correlation analysis shows the immune factors and drug responses associated with IKZF3 expression. In conclusion, the present study is the first, to our knowledge, to identify a pan-cancer genomic signature of the Ikaros gene family among different cancers. Expression of these family members, particularly high levels of IKZF3, indicate positive immunological status and beneficial clinical outcomes of SKCM. IKZF3 may therefore serve as potential targets for immunotherapy of melanoma.

Value of Contrast-enhanced Ultrasound in Evaluating Synovitis and Predicting Recurrent Joint Bleeding of Hemophilia.

OBJECTIVE: Contrast-enhanced ultrasound (CEUS) is advantageous for evaluating microcirculation, and has been applied to assess arthritis in previous studies. However, CEUS examinations have not been studied for hemophilia arthritis. Hemophilia arthritis is different from other arthritis, because it is induced by spontaneous joint bleeding. Hence, CEUS may have special value in evaluating hemophilia arthritis. The present study assessed the value of CEUS in evaluating synovial hypertrophy and predicting recurrent joint bleeding in severe hemophilia A patients. METHODS: From August 2016 to January 2017, 81 severe hemophilia A patients, who were referred to our hospital for ultrasound joint assessment with conventional ultrasound, were enrolled. Among these 81 patients, 46 patients consented for CEUS examinations on the same day. RESULTS: Compared to color Doppler flow imaging (CDFI), four more joints presented with a blood flow signal under CEUS mode. In addition, the synovial hypertrophy measured by CEUS was thicker than that measured by conventional ultrasound. The ultrasound scores (including the total grey-scale ultrasound score, joint effusion/hemarthrosis, synovial hypertrophy, CDFI semi-quantitative score, and CEUS semi-quantitative score) were significantly higher in the joint bleeding group than in the no joint bleeding group (P<0.05). Furthermore, these ultrasound scores were positively correlated with the joint bleeding frequency, and had the highest correlation with the CEUS score (r=0.620, P<0.05). CONCLUSION: CEUS can more accurately assess the degree of synovial hypertrophy and vascularization, and diagnose synovitis, when compared to conventional ultrasound. In addition, CEUS appears to be essential for evaluating the possibility of recurrent joint bleeding, and providing more reliable evidence for individualized treatment.

Autologous fat transplantation for the treatment of abdominal wall scar adhesions after cesarean section.

Cesarean section results in scarring, which usually leads to adhesion between the subcutaneous fat and the abdominal wall muscle. The present study aimed to evaluate the therapeutic effect of autologous fat grafting on scar adhesion to the abdominal wall after cesarean section. Thirty-six patients with scar adhesion to the abdominal wall after cesarean section were recruited and treated between October 2013 and December 2015. The adhesion between the subcutaneous fat and the abdominal wall muscle was carefully separated through a small incision in the original scar to form multiple subcutaneous tunnels. Aspirated fat was injected into the scar lesion and subcutaneous tunnels, and the wound was then sutured. The clinical outcome was evaluated by comparing the pretreatment and 1-year posttreatment photographs and Patient and Observer Scar Assessment Scale (POSAS) scores. All patients had a marked improvement in the appearance, texture, and depression of the scar during 12 months of follow-up. The 1-year posttreatment POSAS scores for the color, pain, pruritus, hardness, fullness, mobility, and appearance of the scar were significantly decreased compared with the pretreatment scores. Hematoxylin-eosin staining revealed adipocyte-like cells in treated scar tissue specimens obtained 1 year after treatment. None of the patients reported severe adverse reactions. Autologous fat grafting combined with adhesion release may be a good treatment option for abdominal wall scarring after cesarean section. This method is minimally invasive and effective in achieving good functional and esthetic outcomes.

Antiaging Properties of Exosomes from Adipose-Derived Mesenchymal Stem Cells in Photoaged Rat Skin.

Adipose-derived stem cells (ADSCs) have been documented as possible candidates for skin rejuvenation. However, the effects of ADSC-derived exosomes on photoaged skin remain to be fully elucidated. This study was aimed at determining the antiaging effects of ADSC-derived exosomes on photoaged skin. Human ADSCs were isolated from the adipose tissue of healthy women and cultured in vitro. Then, exosomes were extracted from the cultured ADSCs, purified by ultracentrifugation, and verified by examination of cell morphology using transmission electron microscopy and the identification of specific biomarkers. Meanwhile, the optimal exosome concentration and treatment time were selected. The photoaged skin model was created by subjecting Sprague-Dawley rats to ultraviolet B radiation. Exosomes were injected into the photoaged skin in a single therapeutic dose. The thickness of the epidermis and dermis was observed by HE staining. The relative mRNA expression of type I collagen, type III collagen, and matrix metalloproteinases (MMP-1 and MMP-3) was determined by real-time PCR. In the rat model of photoaged skin, the injected exosomes markedly decreased the epidermal thickness and increased the dermal thickness of the photoaged skin 7 days after treatment. Moreover, the proportion of the stratum corneum of the epidermis was decreased. Furthermore, real-time RT-PCR showed that the mRNA expression of type I collagen was increased and that of type III collagen, MMP-1, and MMP-3 was decreased. Our results demonstrate that ADSC-derived exosome treatment could significantly improve skin photodamage and that ADSC-derived exosomes may be a potential agent for photoaged skin treatment.

Allogeneic Platelet-Rich Plasma Therapy as an Effective and Safe Adjuvant Method for Chronic Wounds.

BACKGROUND: Platelet-rich plasma (PRP) improves the healing of refractory wounds, and its application is receiving more attention in the field of wound repair. However, when a patient's condition is very poor, it may be difficult to provide whole blood to harvest autologous PRP. METHODS: We evaluated the efficacy and safety of allogeneic PRP in the field of chronic refractory wound repair. Sixty patients (39 males and 21 females, 57 ± 10 y old) with chronic wounds were enrolled in this prospective, randomized, single-center study during January 2014 to January 2018. Their wounds were treated by standard care. The patients with chronic refractory wounds were divided into allogeneic PRP treatment and control groups on the basis of the presence or absence of allogeneic PRP in wounds after debridement, respectively. Allogeneic PRP was prepared by collecting whole blood from healthy individuals and two-step centrifugation. Clinical effects were evaluated by visually observing wound conditions and objectively assessing wound surfaces. RESULTS: After 30 d of treatment, the allogeneic PRP-treated group showed bright red granulation that bled easily with reduced inflammatory exudation. No rejection reactions were observed. The rate of chronic wound healing was much faster in the allogeneic PRP-treated group than that in the control group. CONCLUSIONS: The present study shows that combined treatment of chronic wounds by standard care and allogeneic PRP significantly shortens healing time, suggesting that allogeneic PRP is an effective, safe adjuvant treatment for chronic wounds.

Critical role of the endogenous renin-angiotensin system in maintaining self-renewal and regeneration potential of epidermal stem cells.

A tightly controlled activity of renin-angiotensin system (RAS) including renin, angiotensin-converting enzymes (ACEs), and angiotensin II (Ang II) receptors is critical not only for maintaining systemic hemodynamics and blood volume but also for controlling cell proliferation, differentiation, and tissue remodeling in target organs. ACE inhibitors or Ang II receptor type 1 (AT1R) blockers are widely used as first line drugs for the treatment of cardiovascular diseases that are caused by chronical activation of RAS. However, about 15% of patients using ACE inhibitors develop side effects in the skin and the underlying mechanisms have been poorly understood or even neglected. Herein we show an endogenous RAS in maintaining self-renewal and regeneration potential of epidermal stem cells (ESCs) thereby contributing to wound healing. Firstly, we found that ESCs may express ACE, and its members in wound edges were positively associated with wound healing in Captopril-treated rats. Secondly, we demonstrated that human ESCs had a functional RAS including ACE1, ACE2, Ang II, AT1R, and AT2R. ACE-Ang II axis maintains human ESC function via activation of both AT1R and AT2R, which are negatively regulated by each other. Ang II-induced activation of extracellular signal-regulated kinase (ERK) and signal transducers and activators of transcription (STAT)1 and STAT3 was mediated by the negative cross-talk between AT1R and AT2R in human ESCs. These results suggest that Ang II is a critical regulator of ESC function and ESC-mediated epidermal regeneration. Inappropriate interruption of Ang II-operated signaling may prejudice ESC function leading to impaired skin wound healing or even disease.

Experimental Study on Effects of Adipose-Derived Stem Cell-Seeded Silk Fibroin Chitosan Film on Wound Healing of a Diabetic Rat Model.

BACKGROUND: Wound healing is a complex process that relies on growth factors and stimulation of angiogenesis. Tissue engineering materials composed of adipose-derived stem cells (ADSCs) and silk fibroin (SF)/chitosan (CS) may be able to solve this problem. The aim of this study was to investigate the wound-healing potential of ADSC-seeded SF/CS in streptozotocin-induced diabetic rats. MATERIALS AND METHODS: Thirty-six male Sprague-Dawley rats were purchased and randomly assigned into 3 groups: a control group (no graft), a group treated with SF/CS film graft, and a group treated with ADSC-seeded SF/CS graft. The number of animals in each group was 12. Diabetes was induced by an intraperitoneal injection of streptozotocin. A cutaneous wound was incised at the dorsal region of all the experimental animals. The ADSCs were labeled with CM-Dil fluorescent staining. Wound healing was assessed for all animal groups by observing the rate of wound closure and hematoxylin and eosin staining. The expression of epidermal growth factor, transforming growth factor-ß, and vascular endothelial growth factor at the wound sites was studied by enzyme-linked immunosorbent assay to evaluate the effect of growth factors secreted by ADSCs. The differentiation of ADSCs was analyzed by immunofluorescence staining. RESULTS: The ADSC-seeded SF/CS film treatment significantly increased the rates of wound closure in treated animals, and hence wound healing was drastically enhanced for ADSC-SF/CS treatment groups compared with control groups and SF/CS film treatment group. Histological observations showed the condition of wound healing. Enzyme-linked immunosorbent assay and immunofluorescence staining observations showed the secretion and differentiation of ADSCs, respectively. CONCLUSIONS: Our analyses clearly suggested that it is feasible and effective to enhance wound healing in a diabetic rat model with ADSC-seeded SF/CS film.

Preconditioning With Low-Level Laser Irradiation Enhances the Therapeutic Potential of Human Adipose-derived Stem Cells in a Mouse Model of Photoaged Skin.

This study was conducted to explore the therapeutic potential of human adipose-derived stem cells (ADSCs) irradiated with a low-level laser (LLL). Cultured ADSCs were treated with 650-nm GaAlAs laser irradiation at 2, 4 and 8 J cm-2 . Cell proliferation was quantified by MTT assays, cytokine secretion was determined by enzyme-linked immunosorbent assays, and adipogenic differentiation was examined by oil red O staining. Additionally, the expression profiles of putative ADSC surface markers were analyzed by quantitative real-time PCR. In addition, a mouse photoaged skin model was established by UVB irradiation. Effects of GaAlAs laser-treated ADSCs on the thicknesses of the epidermis and dermis were analyzed by hematoxylin and eosin staining. The results showed that GaAlAs laser treatment of cells at a radiant exposure of 4 J cm-2 enhanced ADSC proliferation and adipogenic differentiation and increased secretion of growth factors. Furthermore, GaAlAs laser irradiation upregulated the expression of putative ADSC surface markers. In the mouse model of photoaged skin, ADSCs treated with GaAlAs laser irradiation had markedly decreased the epidermal thickness and increased the dermal thickness of photoaged mouse skin. Our data indicate that LLL irradiation is an effective biostimulator of ADSCs and might enhance the therapeutic potential of ADSCs for clinical use.

Improved fat transplantation survival by using the conditioned medium of vascular endothelial growth factor transfected human adipose-derived stem cells.

Autologous fat transplantation has been applied widely in clinic. However, the low survival rate is still a problem to be solved. Studies shows that the human adipose-derived stem cells (ADSCs) transfected by vascular endothelial growth factor (VEGF) can improve the survival rate of autologous fat transplantation. Our study is to evaluate the effects of the conditioned medium of VEGF-transfected human adipose-derived stem cells (VEGF-ADSCs-CM) on fat transplantation. ADSCs were isolated and transfected with MOI = 40. The study was divided into three groups, VEGF-ADSCs-CM group, normal-ADSCs-CM group and control group. The conditioned media for VEGF-ADSCs-CM group and normal-ADSCs-CM group were collected, and then mixed with fat, with the mixtures being injected into the back of nude mice. On 4, 7, 15, 30, 60 days after transplantation, the grafts were evaluated on the wet weight, histology, ELISA and western blot. As the results revealed, the survival rate of VEGF-ADSCs-CM group was highest with the best fat cell morphology, and the VEGF secretion of VEGF-ADSCs-CM group was also highest. Therefore, our study demonstrates that VEGF-ADSCs-CM can improve the survival rate of fat transplantation effectively, and VEGF-ADSCs-CM can be regarded as an effective assisted method for fat transplantation.

Facial intramuscular lipoma occurrence following topical cosmetic injection with a mixture of basic fibroblast growth factor: A report of two cases.

Growth factors and cytokines control cell growth, proliferation and differentiation via a network of inter- and intracellular signalling pathways, and are involved in skin self-renewing and wound healing. In recent years, topical and injectable growth factors and cytokines have emerged as an intriguing therapeutic modality that can be harnessed for aesthetic purposes. However, very little data are available on their long-term safety and tolerability. In this report, we describe two cases of patients, who developed intramuscular lipoma of the chin following topical injection with a mixture of basic fibroblast growth factor as the main ingredients for chin augmentation. Biopsies in the two cases were performed at our department, and revealed intramuscular lipoma. Our report indicates that the topical injection of growth factors can lead to tumorigenesis, so health care providers need to be aware of its potential consequences.

Evaluation of 2 Purification Methods for Isolation of Human Adipose-Derived Stem Cells Based on Red Blood Cell Lysis With Ammonium Chloride and Hypotonic Sodium Chloride Solution.

BACKGROUND: The present study was conducted to compare 2 purification methods for isolation of human adipose-derived stromal vascular fraction or stem cells (ADSCs) based on red blood cell (RBC) lysis with 155 mM ammonium chloride (NH4Cl) and hypotonic sodium chloride (NaCl) solution, and try to develop a safe, convenient, and cost-effective purification method for clinical applications. METHODS: Adipose-derived stem cells and RBC were harvested from the fatty and fluid portions of liposuction aspirates, respectively. The suitable concentration of hypotonic NaCl solution on RBC lysis for purification of ADSCs was developed by RBC osmotic fragility test and flow cytometry analysis. The effects of 155 mM NH4Cl or 0.3% NaCl solution on ADSCs proliferation and RBC lysis efficiency were examined by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide assay and lysis efficiency test, respectively. In addition, the adipogenic and osteogenic capabilities, phenotype and genetic stability of ADSCs were evaluated by oil red staining, alkaline phosphatase activity measurement, flow cytometry, and karyotype analysis, respectively. RESULTS: Sodium chloride solution in 0.3% concentration effectively removed RBCs and did not influence the survival of ADSCs in the 10-minute incubation time. The lysis efficiency did not differ significantly between 0.3% NaCl and 155 mM NH4Cl. Moreover, the adipogenic and osteogenic capabilities, surface marker expression and karyotype of the ADSCs were not affected by lysis solutions or by lysis per se. However, the proliferation capacity in the 0.3% NaCl group was superior to that in 155 mM NH4Cl group. CONCLUSIONS: Our data suggest that 0.3% NaCl solution is useful for isolating ADSCs from liposuction aspirate for clinical applications with safety, convenience, and cost-effect.

Effect of suction pressures on cell yield and functionality of the adipose-derived stromal vascular fraction.

OBJECTIVE: The present study was performed to explore the effect of suction pressures on the cell yield of adipose-derived stromal vascular fraction (SVF) and the functionality of adipose-derived stem cells (ADSCs), situated in the SVF, and to develop optimal parameters of harvesting SVF for clinical use. METHODS: Adipose tissue was harvested from the lower abdomen of 10 patients by suction-assisted lipoplasty. Suction pressure was either -30 ± 5 kPa or -55 ± 5 kPa. The aspirated samples were subjected to macroscopic observation to verify the adipose particle size and cytological analysis to detect the cell yield and functionality of the SVF harvested. RESULTS: Adipose tissue harvested at -30 ± 5 kPa appeared to have smaller particle sizes and less blood red cells than that harvested at -55 ± 5 kPa. Cell counts revealed that the cell number of the SVF obtained at -30 ± 5 kPa was more than 2-fold higher than that obtained at -55 ± 5 kPa. Cell growth at passages 1 and 2 was faster at -30 ± 5 kPa than that at -55 ± 5 kPa. The secretion of basic fibroblast growth factor and vascular endothelial growth factor as well as the capacity for adipogenic differentiation of the cultured cells at passages 1-3 were higher at -30 ± 5 kPa than those at -55 ± 5 kPa. There was no difference in the expression of the phenotypic markers between the two groups. CONCLUSION: Our data indicate that the pressure for harvesting adipose tissue affects the yield and viability of the SVF. A lower suction pressure is beneficial to harvesting the SVF for clinical use.

Combined alloplastic implant and autologous dermis graft for nasal augmentation rhinoplasty in Asians.

UNLABELLED: Alloplastic implants may be used in augmentation rhinoplasty but are associated with thinning of the skin over the implant as well as extrusion and translucency of the implant. To minimize these complications, this report describes a combined alloplastic implant and autologous dermal graft for dorsal and tip augmentation rhinoplasty. Of 37 Chinese patients, 35 (94.6 %) were satisfied with the outcome of this procedure during a follow-up period of up to 24 months, and no implant extrusions occurred. The preliminary findings indicate that a combined alloplastic implant and autologous dermis graft is appropriate for nasal augmentation, especially for patients with thin tip skin. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Helium-neon laser irradiation promotes the proliferation and migration of human epidermal stem cells in vitro: proposed mechanism for enhanced wound re-epithelialization.

OBJECTIVE: The present study was conducted to investigate the effects of helium-neon (He-Ne) laser irradiation on the proliferation, migration, and differentiation of cultured human epidermal stem cells (ESCs). BACKGROUND DATA: A He-Ne laser with a wavelength of 632.8 nm is known to have photobiological effects, and is widely used for accelerating wound healing; however, the cellular mechanisms involved have not been completely understood. METHODS: The ESCs were prepared from human foreskin, and irradiated by using He-Ne laser at 632.8 nm with 2 J/cm(2). The ESC proliferation, migration, and differentiation were examined by using XTT assay, scratch assay, and flow cytometry technology, respectively. The phosphorylation of extracellular signal-regulated kinases (ERK) was analyzed by using Western blotting. RESULTS: He-Ne laser irradiation markedly promoted cell proliferation and migration accompanied by an increase in the phosphorylation of ERK, but did not significantly influence cell differentiation. CONCLUSION: Our data indicated that photostimulation with a He-Ne laser resulted in a significant increase in human ESC proliferation and migration in vitro, which might contribute, at least partially, to accelerated wound re-epithelialization by low-level laser therapy.

[The change of angiotensin II production and its receptor expression during wound healing: possible role of angiotensin II in wound healing].

OBJECTIVE: This study was undertaken to observe the change in the local level of angiotensin II (Ang II) and the expression of its corresponding receptors AT1 and AT2 during wound healing, and explore the possible role of Ang II in wound healing . METHODS: A model of full-thickness cutaneous wound was developed on the back of C57/BL6 mice. Specimens were taken from the wound of each mouse on the day 0, 1, 3, 5, 7, 9, 11, 13 and 15 after wounding. The change in the generation of Ang II in wounded tissue during the healing process was detected with ELISA. The proliferation and the apoptosis of cells were detected by bromodeoxyuridine (Brdu) and terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling (TUNEL) method in wounded skin during the healing process, respectively. The cellular localization and the mRNA level change of Ang II receptors in wounded tissue during healing were detected with immunostaining and RT-PCR. RESULTS: Ang II produced in wounded skin was increased in the first 7 days to reach the peak, and then gradually decreased during wound healing. BrdU labeling index was increased gradually in the first 7 days to reach the peak, and then gradually decreased during wound healing. The number of TUNEL-positive cells was increased slowly in the first 7 days after wounding. The increase in the number of TUNEL-positive cells was more markedly after epithelization of the wound. In normal mice, AT1 and AT2 receptor were found positively expressed in the whole epidermal layer, while positive expression was only found in the endothelial cells of the capillary vessels within the dermal layer, and positive expression was also found in appendages of the skin, i. e. hair follicle, sweat gland and sebaceous gland respectively. Positive staining signal of both AT1 and AT2 receptors were increased in the first 7 days to reach the peak, then gradually decreased. Expression of AT2R was increased again following the epithelization of wound. The result of RT-PCR showed that the expression of both AT1 and AT2 receptors was detectable, and AT1 receptor was increased in the first 7 days to the peak, and then gradually decreased during wound healing, while AT2 receptor expression reached its peak value on day 7, then gradually decreased, and increased again following the epithelization of wound. CONCLUSIONS: These results indicate that Ang II participate in wound repair and related to remolding in the late stage of wound healing through the change in production of angiotensin II and expression of AT1 and AT2 receptors. AT1 receptor might be closely associated with cell proliferation, while AT2 receptor might play a role in cell apoptosis and remolding during wound healing.