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BACKGROUND: THOC7-AS1 and FSTL1 expression are frequently upregulated in cutaneous squamous cell carcinoma (cSCC). However, their molecular biological mechanisms remain elusive and their potential as therapeutic targets needs urgent exploration. METHODS: Human tissue samples were used to evaluate clinical parameters. In vitro and in vivo experiments assessed biological functions. Quantitative PCR, western blot, immunohistochemistry, immunocytochemistry, immunoprecipitation, RNA fluorescence in situ hybridization, RNA pull-down, RNA immunoprecipitation, silver staining, chromatin immunoprecipitation, dual luciferase reporter assays etc. were utilized to explore the molecular biological mechanisms. RESULTS: We found FSTL1 is an oncogene in cSCC, with high expression in tumor tissues and cells. Its elevated expression closely associates with tumor size and local tissue infiltration. In vitro and in vivo, high FSTL1 expression promotes cSCC proliferation, migration and invasion, facilitating malignant behaviors. Mechanistically, FSTL1 interacts with ZEB1 to promote epithelial-to-mesenchymal transition (EMT) in cSCC cells. Exploring upstream regulation, we found THOC7-AS1 can interact with OCT1, which binds the FSTL1 promoter region and promotes FSTL1 expression, facilitating cSCC progression. Finally, treating tumors with THOC7-AS1 antisense oligonucleotides inhibited cSCC proliferative and migratory abilities, delaying tumor progression. CONCLUSIONS: The THOC7-AS1/OCT1/FSTL1 axis regulates EMT and promotes tumor progression in cSCC. This study provides clues and ideas for cSCC targeted therapy.
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Carcinoma de Células Escamosas , Proteínas Relacionadas con la Folistatina , Neoplasias Cutáneas , Humanos , Carcinoma de Células Escamosas/patología , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Proteínas Relacionadas con la Folistatina/genética , Proteínas Relacionadas con la Folistatina/metabolismo , Regulación Neoplásica de la Expresión Génica , Hibridación Fluorescente in Situ , ARN , ARN Largo no Codificante/genética , Neoplasias Cutáneas/genética , Neoplasias Cutáneas/patologíaRESUMEN
Cutaneous squamous cell carcinoma (cSCC) ranks as the second most prevalent skin tumour (excluding melanoma). However, the molecular mechanisms driving cSCC progression remain elusive. This study aimed to investigate GBP1 expression in cSCC and elucidate its potential molecular mechanisms underlying cSCC development. GBP1 expression was assessed across public databases, cell lines and tissue samples. Various assays, including clone formation, CCK8 and EdU were employed to evaluate cell proliferation, while wound healing and transwell assays determined cell migration and invasion. Subcutaneous tumour assays were conducted to assess in vivo tumour proliferation, and molecular mechanisms were explored through western blotting, immunofluorescence and immunoprecipitation. Results identified GBP1 as an oncogene in cSCC, with elevated expression in both tumour tissues and cells, strongly correlating with tumour stage and grade. In vitro and in vivo investigations revealed that increased GBP1 expression significantly enhanced cSCC cell proliferation, migration and invasion. Mechanistically, GBP1 interaction with SP1 promoted STAT3 activation, contributing to malignant behaviours. In conclusion, the study highlights the crucial role of the GBP1/SP1/STAT3 signalling axis in regulating tumour progression in cSCC. These findings provide valuable insights into the molecular mechanisms of cSCC development and offer potential therapeutic targets for interventions against cSCC.
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Carcinoma de Células Escamosas , Movimiento Celular , Proliferación Celular , Proteínas de Unión al GTP , Invasividad Neoplásica , Factor de Transcripción STAT3 , Neoplasias Cutáneas , Factor de Transcripción Sp1 , Factor de Transcripción STAT3/metabolismo , Humanos , Neoplasias Cutáneas/metabolismo , Neoplasias Cutáneas/patología , Neoplasias Cutáneas/genética , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patología , Carcinoma de Células Escamosas/genética , Factor de Transcripción Sp1/metabolismo , Proteínas de Unión al GTP/metabolismo , Proteínas de Unión al GTP/genética , Línea Celular Tumoral , Animales , Ratones , Transducción de Señal , Femenino , Ratones DesnudosRESUMEN
OBJECTIVE: To assess the safety and efficacy of antimicrobial peptide PL-5 (Peceleganan) spray in the treatment of wound infections. BACKGROUND: Antimicrobial peptide PL-5 spray is a novel topical antimicrobial agent. METHODS: We conducted a multicenter, open-label, randomized, controlled phase IIb clinical trial to evaluate the efficacy and safety of PL-5 spray, as compared with silver sulfadiazine, in patients with skin wound infections. The primary efficacy outcome was the clinical efficacy rate on the first day after ending the treatment (D8). The secondary efficacy outcome was the clinical efficacy rate on the fifth day posttreatment (D5), the bacteria clearance rate, and the overall efficacy rate at the mentioned 2 time points. The safety outcomes included adverse reactions and pharmacokinetic analysis posttreatment. RESULTS: A total of 220 patients from 27 hospitals in China were randomly assigned to 4 groups. On D8, the efficacy rate was 100.0%, 96.7%, 96.7% for the 1 PL-5, 2 PL-5, 4 PL-5 groups, respectively, as compared with 87.5% for the control group. The efficacy rate among the 4 groups was significantly different ( P <0.05). On D5, the efficacy rate was 100.0%, 93.4%, 98.3% for the 1 PL-5, 2 PL-5, 4 PL-5 groups, respectively, as compared with 82.5% for the control group. The efficacy rate among the 4 groups was significantly different ( P <0.05). The blood concentration of PL-5 was not detectable in pharmacokinetic analysis. No severe adverse event related to the application of PL-5 was reported. CONCLUSIONS: Antimicrobial peptide PL-5 spray is safe and effective for the treatment of skin wound infections. TRIAL REGISTRATION: ChiCTR2000033334.
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Antiinfecciosos Locales , Infección de Heridas , Humanos , Resultado del Tratamiento , Bacterias , China , Método Doble CiegoRESUMEN
Thermal injury repair is a complex process during which the maintenance of the proliferation and migration of human skin fibroblasts (HSFs) exert a crucial role. MicroRNAs have been proven to exert an essential function in repairing skin burns. This study delves into the regulatory effects of miR-24-3p on the migration and proliferation of HSFs that have sustained a thermal injury, thereby, providing deeper insight into thermal injury repair pathogenesis. The PPAR-ß protein expression level progressively increased in a time-dependent manner on the 12th, 24th and 48th hour following the thermal injury of the HSFs. The knockdown of PPAR-ß markedly suppressed the proliferation of and migration of HSF. Following thermal injury, the knockdown also promoted the inflammatory cytokine IL-6, TNF-α, PTGS-2 and P65 expression. PPAR-ß contrastingly exhibited an opposite trend. A targeted relationship between PPAR-ß and miR-24-3p was predicted and verified. miR-24-3p inhibited thermal injured HSF proliferation and migration and facilitated inflammatory cytokine expression through the regulation of PPAR-ß. p65 directly targeted the transcriptional precursor of miR-24 and promoted miR-24 expression. A negative correlation between miR-24-3p expression level and PPAR-ß expression level in rats' burnt dermal tissues was observed. Our findings reveal that miR-24-3p is conducive to rehabilitating the denatured dermis, which may be beneficial in providing effective therapy of skin burns.
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Quemaduras , MicroARNs , PPAR-beta , Animales , Quemaduras/genética , Proliferación Celular , Citocinas/metabolismo , Fibroblastos/metabolismo , Humanos , MicroARNs/genética , MicroARNs/metabolismo , FN-kappa B/metabolismo , PPAR-beta/genética , PPAR-beta/metabolismo , RatasRESUMEN
Our previous study confirmed the critical role of miR-125b and vascular endothelial growth factor (VEGF) in burn wound repair., The present study was aimed to identify the role of long noncoding RNAs (lncRNAs) related to the function of miR-125b and VEGF in burn wound repair and the underlying mechanism. First, we found that lncRNA PDK1-AS and VEGFA expression was significantly increased in heat-denatured dermal tissue samples and in human dermal microvascular endothelial cells (HDMECs) and human umbilical vein endothelial cells (HUVECs) after thermal injury. PDK1-AS knockdown significantly inhibited cell viability, cumulative tube length, cell migratory ability, and cell invasion of thermally injured HDMECs and HUVECs. PDK1-AS knockdown decreased VEGFA protein levels in HDMECs and HUVECs. While overexpression of PDK1-AS showed the opposite effects. Online tools prediction and luciferase assay confirmed that miR-125b-5p targeted PDK1-AS and VEGFA 3'-untranslated region. miR-125b-5p inhibition significantly increased VEGFA protein levels and enhanced viability, cumulative tube length, migratory ability, and invasion of HUVECs and HDMECs. Furthermore, the effects of PDK1-AS knockdown on VEGFA protein levels in the two cell lines were partially reversed by miR-125b-5p inhibition. Finally, in the tissue samples, PDK1-AS and VEGFA expression was increased, while miR-125b-5p expression was decreased in heat-denatured dermal tissues; the expression of miR-125b-5p had a negative correlation with PDK1-AS and VEGFA, respectively, and PDK1-AS and VEGFA were positively correlated with each other in tissue samples. In conclusion, PDK1-AS relieves miR-125b-5p-induced inhibition on VEGFA by acting as a endogenous RNA, therefore modulating HDMEC and HUVEC angiogenesis after thermal injury.
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Dermis/irrigación sanguínea , Células Endoteliales de la Vena Umbilical Humana/metabolismo , MicroARNs/metabolismo , Microvasos/patología , Neovascularización Fisiológica , ARN Largo no Codificante/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Regiones no Traducidas 3'/genética , Secuencia de Bases , Quemaduras/genética , Quemaduras/patología , Regulación de la Expresión Génica , Humanos , MicroARNs/genética , Neovascularización Fisiológica/genética , ARN Largo no Codificante/genéticaRESUMEN
BACKGROUND: Slow lymphangiogenesis is one crucial reason for the impaired wound healing process in diabetes. Accumulative evidence showed that long noncoding RNA-antisense noncoding RNA in the INK4 locus (ANRIL) could influence lymphangiogenesis. Besides, miR-181a has been reported to regulate Prox1 that is essential for lymphangiogenesis. However, the relationship between ANRIL and miR-181a as well as the definitive function of ANRIL in lymphangiogenesis is not clear. METHODS: The diabetic mouse model was set up to assess the wound healing rate in vivo. Quantitative real-time polymerase chain reaction was performed to measure the expressions of ANRIL, miR-181a, and Prox1. Western blot analysis was used to assess the expressions of vascular endothelial growth factor receptor-3, lymphatic vessel hyaluronan receptor-1, Prox1, and epithelial-mesenchymal transition (EMT)-related proteins. Flow cytometry was used to assess the cell apoptosis. Wound healing assay was used to determine the effect of ANRIL on cell migration. Tube-formation assay and immunofluorescence staining were performed to determine tube-formation capacity of human dermal lymphatic endothelial cells (LECs). RESULTS: ANRIL and Prox1 were downregulated, whereas miR-181a was upregulated in the diabetic wound healing mouse model and high glucose (HG)-induced LECs. The wound healing rate and EMT were inhibited during the diabetic wound healing process. Dual-luciferase assay proved that miR-181a could bind Prox1 to repress its expression, whereas ANRIL could sponge miR-181a to recover Prox1 expression. Overexpression of ANRIL or inhibition of miR-181a rescued the impairments of survival, migration, EMT formation, and tube formation of LECs caused by HG. CONCLUSION: ANRIL could promote lymphangiogenesis during the diabetic wound healing process via sponging miR-181a to enhance Prox1 expression, which might help design new therapy to improve the wound healing efficacy for diabetes.
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Complicaciones de la Diabetes/metabolismo , Diabetes Mellitus Experimental/metabolismo , Proteínas de Homeodominio/metabolismo , Linfangiogénesis , Vasos Linfáticos/metabolismo , MicroARNs/metabolismo , ARN Largo no Codificante/metabolismo , Úlcera Cutánea/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Cicatrización de Heridas , Animales , Glucemia/metabolismo , Movimiento Celular , Células Cultivadas , Complicaciones de la Diabetes/genética , Complicaciones de la Diabetes/patología , Complicaciones de la Diabetes/fisiopatología , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patología , Diabetes Mellitus Experimental/fisiopatología , Transición Epitelial-Mesenquimal , Regulación de la Expresión Génica , Proteínas de Homeodominio/genética , Humanos , Vasos Linfáticos/patología , Vasos Linfáticos/fisiopatología , Ratones Endogámicos C57BL , MicroARNs/genética , ARN Largo no Codificante/genética , Transducción de Señal , Úlcera Cutánea/genética , Úlcera Cutánea/patología , Factores de Tiempo , Proteínas Supresoras de Tumor/genéticaRESUMEN
OBJECTIVE: To summarize the clinical features of Marjolin's ulcers in lower limbs and the diagnosis and treatment methods for it.â© Methods: The clinical data of 89 patients with lower limbs Marjolin's ulcers, who were treated in Xiangya Hospital, Central South University from Jan 1998 to Dec 2017, were retrospectively analyzed, including demographics, injury factors, length of cancer incubation period, lesion location, ulcer area, pathological type, bone invasion, lymph node metastasis, surgical methods, repair methods and prognosis.â© Results: There were 70 males and 19 females among 89 patients with lower limbs Marjolin's ulcers. The most common injuries were flame burn (42 cases), trauma (19 cases), and burns (12 cases). The lesions were most common in the lower leg (31 cases), followed by the thigh (11 cases) and the heel (11 cases). The ulcer area was 1.5-600.0 cm2. There were 80 cases of squamous cell carcinoma, 8 cases of verrucous carcinoma, and 1 case of sarcoma. Before operation, 78 cases of inguinal lymphadenectasis were found, 49 cases of inguinal lymph node dissection, 29 cases of simple lymph node biopsy and resection, and 9 cases of lymph node metastasis and 8 cases of bone invasion were observed; 24 cases of amputation, 53 cases of extended resection and skin grafts, and 12 patients of extensive resection and flap transplantation were performed. Sixty-five cases were followed up, and 8 cases recurred, including 2 cases of amputation patients and 6 cases of extended resection patients. There was no relationship between recurrence of tumors and surgical methods (P>0.05).â© Conclusion: The recurrence and metastasis rate of Marjolin's ulcers in lower limbs is high, requiring early detection, early diagnosis, early surgical treatment and regular follow-up. Lnguinal lymphadenectasis is more common and requires lymph node biopsy and lymphadenectomy, or lymph node dissection. Extended local resection, skin graft or flap repair is the main treatment methods. However, amputation can be considered if the cancer is big, the invasion is deep, and the lower extremity scar is extensive and combined with severe deformity.
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Quemaduras , Carcinoma de Células Escamosas , Neoplasias Cutáneas , Úlcera Cutánea , Femenino , Humanos , Extremidad Inferior , Masculino , Recurrencia Local de Neoplasia , Estudios Retrospectivos , ÚlceraRESUMEN
Denatured dermis is a part of the dermis in deep burn wound and has the ability to restore normal morphology and function. In our previous study, we revealed that miR-29a downregulation in denatured dermis may help burn wound healing in the later phase, and further enhance type I collagen synthesis. LIN28A, a highly-conserved RNA binding protein expressed during embryogenesis, plays roles in development, pluripotency, metabolism, as well as tissue repair in adults. In the present study, we investigated the functional roles of LIN28A in human skin fibroblasts (HSFs) and extracellular matrix (ECM), and the interaction between miR-29a and LIN28A. In recent years, long non-coding RNAs have been reported to play a key role in normal development and physiology, as well as in disease development. By using online tools, we screened out several candidate lncRNAs of miR-29a, among which XIST was inversely regulated by miR-29a. XIST, one of the first found cancer-associated lncRNAs, has been frequently reported to play major role in several biological processes. Further, we evaluated the roles and mechanism of XIST in HSF proliferation, migration, and ECM synthesis. Through regulation of miR-29a/LIN28A, XIST knockdown suppressed HSF proliferation, migration, and ECM synthesis. In denatured dermis tissues, XIST, and LIN28A expression was upregulated, miR-29a expression was downregulated. Taken together, promoting XIST expression in denatured dermis, thus to inhibit miR-29a and promote LIN28A expression, further promote HSF proliferation, migration, and ECM synthesis presents a promising strategy for denatured dermis repair.
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Quemaduras/genética , MicroARNs/genética , ARN Largo no Codificante/genética , Proteínas de Unión al ARN/genética , Movimiento Celular , Proliferación Celular , Células Cultivadas , Dermis/citología , Fibroblastos/citología , Células HEK293 , Calor , HumanosRESUMEN
BACKGROUND Marjolin ulcer (MU) is an aggressive cutaneous malignancy. Typically, MU occurs over a period of time in post-burn and/or post-traumatic lesions and scars. However, the pathogenesis of scar carcinogenesis and MU development remains to be elucidated. The present study aimed to investigate the long noncoding RNA (lncRNA) and messenger RNA (mRNA) expression profiling in MU, which could provide new information on the potential molecular mechanisms of MU development. MATERIAL AND METHODS The lncRNA microarray analysis was conducted in normal skin, scar, and MU tissue, and quantitative real-time PCR experiment was carried out to validate the reliability of the microarray data. Furthermore, a series of integrative bioinformatic approaches were applied to decipher the function of differentially expressed lncRNAs. RESULTS A total of 7130 lncRNAs and 9867 mRNAs were differentially expressed among normal skin, scar, and MU tissues. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis demonstrated that these aberrantly expressed transcripts were mainly involved in cell cycle, immune response, and the p53 signaling pathway. Series Test of Cluster analysis indicated certain dysregulated lncRNAs were expressed with a gradually increasing or decreasing trend and might participated in malignant transformation of scar tissue postburn. Co-expression analysis showed 5 selected lncRNAs might regulate cell proliferation through the p53 signaling pathway. Finally, the competing endogenous RNA (ceRNA) network indicated that lncRNA uc001oou.3 might be implicated in ceRNA mechanism during MU development. CONCLUSIONS Taken together, our study implied the aberrant expression of lncRNAs may play an important role in the pathogenesis and development of MU, and the exact mechanism warrants further investigation.
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Carcinoma de Células Escamosas/genética , ARN Largo no Codificante/genética , ARN Mensajero/genética , Adulto , Análisis por Conglomerados , Biología Computacional , Femenino , Perfilación de la Expresión Génica/métodos , Ontología de Genes , Redes Reguladoras de Genes/genética , Humanos , Masculino , MicroARNs/genética , Persona de Mediana Edad , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN Largo no Codificante/metabolismo , ARN Mensajero/metabolismo , Análisis de Secuencia de ARN , Transcriptoma/genética , Úlcera/genética , Úlcera/metabolismoRESUMEN
Decreased Th1/Th2 ratio is one of the major characteristics of immunosuppression in sepsis. Both membrane adhesive protein Annexin-A1 (ANXA1) and transcription factor GATA-3 have been reported to play important roles in T cell differentiation. However, the relationship between ANXA1 and GATA-3 in Th1/Th2 shift is unknown. Our study investigated the interaction effects of ANXA1 and GATA-3 to influence T cell differentiation in CD4+ T cells. We found that GATA-3 and ANXA1 were coexpressed on Th0/Th1/Th2 cytoplasm and nuclear. Overexpressed ANXA1 significantly increased the expression of IFNγ and reduced IL-4 expression in T cells, while ANXA1-silenced T cells exhibited decreased production of IFNγ and increased production of IL-4. Knockdown of ANXA1 promoted higher expression level of GATA-3 and low level of T-box transcription factor (T-bet/Tbx21). Further study demonstrated that ANXA1 regulated GATA-3 expression through the formyl peptide receptor like-1 (FPRL-1) downstream signaling pathways ERK and PKB/Akt. These results suggested that ANXA1 modulates GATA-3/T-bet expression induced Th0/Th1 differentiation. Moreover, we found that GATA-3 inhibited ANXA1 expression by binding to its promoter for the first time. It is proposed that the interactions between ANXA1 and GATA-3 may provide clues to understand the immunosuppression and have potential as new therapeutic targets in immunotherapy after sepsis.
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Anexina A1/metabolismo , Linfocitos T CD4-Positivos/inmunología , Factor de Transcripción GATA3/metabolismo , Animales , Anexina A1/genética , Células Cultivadas , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente , Factor de Transcripción GATA3/genética , Terapia de Inmunosupresión , Masculino , Ratones , Unión Proteica , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores de Formil Péptido/genética , Receptores de Formil Péptido/metabolismo , Transducción de Señal/genética , Transducción de Señal/fisiología , TransfecciónRESUMEN
Denatured dermis, a part of dermis in burned skin, has the ability to restore its normal morphology and functions after their surrounding microenvironment is improved. However, the cellular and molecular mechanisms by which the denatured dermis could improve wound healing are still unclear. This study aimed to investigate the role of nucleolin during the recovery of heat-denatured human dermal fibroblasts. Nucleolin mRNA and protein expression were significantly increased time-dependently during the recovery of heat-denatured human dermal fibroblasts (52 °C, 30 seconds). Heat-denaturation promoted a time-dependent cell proliferation, migration, chemotaxis, and scratched wound healing during the recovery of human dermal fibroblasts. These effects were prevented by knockdown of nucleolin expression with small interference RNA (siRNA), whereas overexpression of nucleolin enhanced cell proliferation, migration, and chemotaxis of human dermal fibroblasts with heat-denaturation. In addition, the expression of transforming growth factor-beta 1(TGF-ß1) was significantly increased during the recovery of heat-denatured dermis and human dermal fibroblasts. TGF-ß1 expression was up-regulated by nucleolin in human dermal fibroblasts. The results suggest that nucleolin expression is up-regulated, and play an important role in promoting cell proliferation, migration, and chemotaxis of human dermal fibroblasts during the recovery of heat-denatured dermis with a mechanism probably related to TGF-ß1.
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Quemaduras/fisiopatología , Quimiotaxis/efectos de los fármacos , Dermis/fisiopatología , Fibroblastos/metabolismo , Fosfoproteínas/farmacología , Proteínas de Unión al ARN/farmacología , Factor de Crecimiento Transformador beta1/metabolismo , Cicatrización de Heridas , Western Blotting , Movimiento Celular , Proliferación Celular , Células Cultivadas , Dermis/lesiones , Dermis/metabolismo , Fibroblastos/citología , Regulación de la Expresión Génica , Calor , Humanos , ARN Mensajero , Regulación hacia Arriba , NucleolinaRESUMEN
BACKGROUND: The present study aims to examine the expression patterns and roles of nucleolin during the recovery of heat-denatured human umbilical vein endothelial cells (HUVECs). METHODS: Deep partial thickness burn model in Sprague-Dawley rats and the heat denatured cell model (52°C, 35s) were used. The expression of nucleolin was measured using Western blot analysis and real-time PCR. Angiogenesis was assessed using in vitro parameters including endothelial cell proliferation, transwell migration assay, and scratched wound healing. Gene transfection and RNA interference approaches were employed to investigate the roles of nucleolin. RESULTS: Nucleolin mRNA and protein expression showed a time-dependent increase during the recovery of heat-denatured dermis and HUVECs. Heat-denaturation time-dependently promoted cell growth, adhesion, migration, scratched wound healing and formation of tube-like structures in HUVECs. These effects of heat denaturation on endothelial wound healing and formation of tube-like structures were prevented by knockdown of nucleolin, whereas over-expression of nucleolin increased cell growth, migration, and formation of tube-like structures in cultured HUVEC endothelial cells. In addition, we found that the expression of vascular endothelial growth factor (VEGF) increased during the recovery of heat-denatured dermis and HUVECs, and nucleolin up-regulated VEGF in HUVECs. CONCLUSIONS: The present study reveals that the expression of nucleolin is up-regulated, and plays a pro-angiogenic role during the recovery of heat-denatured dermis and its mechanism is probably dependent on production of VEGF. GENERAL SIGNIFICANCE: We find a novel and important pro-angiogenic role of nucleolin during the recovery of heat-denatured dermis.
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Calor , Fosfoproteínas/fisiología , Proteínas de Unión al ARN/fisiología , Animales , Proliferación Celular , Células Cultivadas , Humanos , Neovascularización Fisiológica/genética , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Proteínas de Unión al ARN/genética , Proteínas de Unión al ARN/metabolismo , Ratas , Ratas Sprague-Dawley , Reacción en Cadena en Tiempo Real de la Polimerasa , Factor A de Crecimiento Endotelial Vascular/metabolismo , NucleolinaRESUMEN
BACKGROUND: Multiple studies have confirmed the mutual enhancement of percutaneous permeation of benzophenone-3 (BP-3) and N,N-diethyl-m-toluamide (DEET), which are effective ingredients in sunscreen products and insect repellents, respectively. However, the association between percutaneous absorption of BP-3 and DEET in a large general adult population remains to be elucidated. METHODS: This cross-sectional study included US adults who had available data on urinary BP-3 and two DEET metabolites, 3-(diethylcarbamoyl) benzoic acid (DCBA) and 3-(ethylcarbamoyl) benzoic acid (ECBA), from the National Health and Nutrition Examination Survey (NHANES) conducted in 2015-2016. We conducted three weighted multivariable linear regression models to investigate the potential correlation between percutaneous absorption of BP-3 and DEET, along with trend tests, smooth curve fitting, and subgroup analysis to assess the robustness of the findings. RESULTS: Weighted multivariable linear logistic regression revealed a positive correlation between log10 BP-3 and log10 DCBA (ß = 0.1678, 95 % CI: 0.0970 to 0.2386) as well as log10 ECBA (ß = 0.1416, 95 % CI: 0.0707 to 0.2125), after adjusting for all covariates. After converting log10 BP-3 from a continuous variable to a categorical variable (quartiles), the trend tests showed that the results were stable (all P for trend < 0.05). Smoothed curve fitting demonstrated a linear positive correlation between log10 BP-3 and both log10 DCBA and log10 ECBA. In subgroup analyses, the positive correlation between BP-3 and DEET metabolites was more pronounced in participants who were male, middle-aged, non-Hispanic white, had a moderate PIR level and reported always or most of the time using sunscreen. CONCLUSIONS: Our findings revealed a statistically significant linear and positive correlation between the percutaneous absorption of BP-3 and DEET in the general adult population.
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Burn injuries, especially severe ones, result in direct and indirect thermal damage to skin tissues, with a complex and slow wound healing process. Improper treatment can induce sustained inflammatory responses, causing systemic damage. Lin28A, a highly conserved RNA binding protein, was found to exert a significant effect on cell proliferation and wound repair. Lin28A exerts the functions through inhibiting the maturation of the let-7 family miRNAs. Herein, the roles of Lin28A and let-7b in thermal injury repair were investigated using a mouse thermal injury model and a human skin fibroblast (HSF) model for thermal injuries. Lin28A could inhibit the maturation of let-7b, thus participating in skin repair after burns. In the animal model, Lin28A was highly expressed after thermal injury. In the HSF model for thermal injuries, downregulation of Lin28A inhibited the proliferation, migration, and extracellular matrix (ECM) generation of fibroblasts. When let-7b was knocked down in HSFs, the impacts on fibroblast functions caused by downregulation of Lin28A were partially reversed. Moreover, let-7b overexpression might significantly attenuate the promotive effects of Lin28A upon thermal injury repair. Finally, AKT2 and IGF1R were the let-7b target genes within cells. These findings reveal that Lin28A might promote thermal injury repair in burn-injured skin by inhibiting the maturation of let-7b and improving HSF viability and functions, thus illustrating the critical effect of let-7b on burn wound healing and providing new therapeutic targets and strategies for burn treatment.
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Quemaduras , Proliferación Celular , Fibroblastos , MicroARNs , Proteínas de Unión al ARN , Piel , Cicatrización de Heridas , Quemaduras/patología , Quemaduras/metabolismo , Quemaduras/genética , Animales , MicroARNs/metabolismo , MicroARNs/genética , Proteínas de Unión al ARN/metabolismo , Proteínas de Unión al ARN/genética , Humanos , Ratones , Fibroblastos/metabolismo , Piel/patología , Piel/metabolismo , Piel/lesiones , Masculino , Movimiento Celular , Ratones Endogámicos C57BL , Modelos Animales de EnfermedadRESUMEN
Importance: Peceleganan spray is a novel topical antimicrobial agent targeted for the treatment of skin wound infections. However, its efficacy and safety remain unclear. Objective: To assess the safety and efficacy of peceleganan spray for the treatment of wound infections. Design, Setting, and Participants: This multicenter, open-label, phase 3 randomized clinical trial recruited and followed up 570 adult patients diagnosed with secondary open wound infections from 37 hospitals in China from August 23, 2021, to July 16, 2022. Interventions: Patients were randomized to 2 groups with a 2:1 allocation. One group received treatment with 2% peceleganan spray (n = 381) and the other with 1% silver sulfadiazine (SSD) cream (n = 189). Main Outcomes and Measures: The primary efficacy outcome was the clinical efficacy rate (the number of patients fulfilling the criteria for efficacy of the number of patients receiving the treatment) on the first day following the end of treatment (day 8). The secondary outcomes included the clinical efficacy rate on day 5 and the bacterial clearance rate (cases achieving negative bacteria cultures after treatment of all cases with positive bacteria cultures before treatment) on days 5 and 8. The safety outcomes included patients' vital signs, physical examination results, electrocardiographic findings, blood test results, and adverse reactions. Results: Among the 570 patients randomized to 1 of the 2 groups, 375 (98.4%) in the 2% peceleganan treatment group and 183 (96.8%) in the 1% SSD control group completed the trial (n = 558). Of these, 361 (64.7%) were men, and the mean (SD) age was 48.6 (15.3) years. The demographic characteristics were similar between groups. On day 8, clinical efficacy was achieved by 339 patients (90.4%) in the treatment group and 144 (78.7%) in the control group (P < .001). On day 5, clinical efficacy was achieved by 222 patients (59.2%) in the treatment group and 90 (49.2%) in the control group (P = .03). On day 8, bacterial clearance was achieved by 80 of 334 patients (24.0%) in the treatment group and in 75 of 163 (46.0%) in the control group (P < .001). On day 5, bacterial clearance was achieved by 55 of 334 patients (16.5%) in the treatment group and 50 of 163 (30.7%) in the control group (P < .001). The adverse events related to the application of peceleganan spray and SSD cream were similar. Conclusions and Relevance: This randomized clinical trial found that peceleganan spray is a safe topical antimicrobial agent with a satisfactory clinical efficacy rate for the treatment of skin wound infections, while the effectiveness of bacterial clearance remains uncertain. Trial Registration: Chinese Clinical Trial Registry Identifier: ChiCTR2100047202.
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Infección de Heridas , Humanos , Masculino , Femenino , Persona de Mediana Edad , Adulto , Infección de Heridas/tratamiento farmacológico , Antiinfecciosos Locales/uso terapéutico , Antiinfecciosos Locales/administración & dosificación , China , Sulfadiazina de Plata/uso terapéutico , Sulfadiazina de Plata/administración & dosificación , Resultado del Tratamiento , Anciano , Antibacterianos/uso terapéutico , Antibacterianos/administración & dosificaciónRESUMEN
Fibrosarcoma is a highly malignant type of soft tissue sarcoma that currently lacks effective treatment options. Polypeptide N-acetylgalactosaminyltransferase 12 (GALNT12) belongs to the uridine diphosphate N-acetylgalactosamine gene family, which is involved in numerous biological processes of diseases, such as tumor progression. Its upregulated expression is closely associated with the development of colorectal cancer. However, research on the role of GALNT12 in fibrosarcoma is currently limited. The present study aimed to assess the expression and biological function of GALNT12 in fibrosarcoma. Patient data and tissue samples were collected and public datasets were obtained from the Gene Expression Omnibus (GSE24369 and GSE21124). Immunofluorescence assays were performed to observe the cellular localization of GALNT12. GALNT12 expression was measured using reverse transcription-quantitative PCR, western blotting and immunohistochemistry. Small interfering RNAs were constructed to knock down GALNT12 expression in HT-1080 cells. Cell Counting Kit-8 and EdU assays were used to assess fibrosarcoma cell proliferation. Wound healing and Transwell assays were used to detect migration. Gene set enrichment analysis was performed to identify key pathways. Paired and unpaired Student's t-test, Fisher's exact test and one-way ANOVA (followed by Tukey's Honest Significant Difference test) were used to analyze the data. It was demonstrated that GALNT12 expression was upregulated in both fibrosarcoma cell lines and tissue samples and predicted poor patient prognosis. In vitro experiments demonstrated that high GALNT12 expression levels significantly increased HT-1080 cell proliferation and migration. Furthermore, it was demonstrated that high GALNT12 expression levels were closely associated with the yes1 associated transcriptional regulator (YAP1) signaling pathway. Knockdown of GALNT12 inhibited YAP1 nuclear translocation, which affected activation of key downstream genes including AMOTL2, BIRC5 and CYR61. Therefore, the present study demonstrated that GALNT12 promoted fibrosarcoma progression. GALNT12 could be a potential biomarker for this disease and may potentially provide new ideas for targeted therapy of fibrosarcoma in the future.
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N6-methyladenosine (m6A) modulates RNA metabolism and functions in cell differentiation, tissue development, and immune response. After acute burns, skin wounds are highly susceptible to infection and poor healing. However, our understanding of the effect of burn injuries on m6A methylation and their potential mechanism is still limited. Human m6A-mRNA&lncRNA Epitranscriptomic microarray was used to obtain comprehensive mRNA and lncRNA transcriptome m6A profiling and gene expression patterns after burn injuries in human skin tissue. Bioinformatic and functional analyses were conducted to find molecular functions. Microarray profiling showed that 65 mRNAs and 39 lncRNAs were significantly hypermethylated; 5492 mRNAs and 754 lncRNAs were significantly hypomethylated. Notably, 3989 hypomethylated mRNAs were down-expressed and inhibited many wound healing biological processes and pathways including in the protein catabolic process and supramolecular fiber organization pathway; 39 hypermethylated mRNAs were up-expressed and influenced the cell surface receptor signaling pathway and inflammatory response. Moreover, we validated that m6A regulators (METTL14, METTL16, ALKBH5, FMR1, and HNRNPC) were significantly downregulated after burn injury which may be responsible for the alteration of m6A modification and gene expression. In summary, we found that homeostasis in the skin was disrupted and m6A modification may be a potential mechanism affecting trauma infection and wound healing.
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BACKGROUND: Diabetes is usually the leading cause of chronic non-healing wounds. LncRNA-GAS5 has been verified to be involved in the regulation of diabetes or high glucose (HG)-stimulated cells. However, its regulatory roles in diabetic wound healing need further investigation. METHOD: GAS5, miR-217 and Prox1 were identified by qRT-PCR. MTT, flow cytometry assay, wound-healing assay and tube formation were used to analyze cell viability, apoptosis, migration and tube formation capacity. Western blotting was carried out to detect the protein expression of c-Myc, CyclinD1, CDK4, Bcl-2, Prox1, VEGFR-3 and LYVE-1. Bioinformatics and luciferase assay were performed to predict and validate the binding sites of miR-217 on GAS5 and Prox1. Immunofluorescence staining detected the expression and distribution of Prox1. The wound healing rate was also assessed by setting up the diabetic mouse model. H&E staining assessed the distribution of inflammatory cells and fibroblasts in the wound tissues. RESULTS: GAS5 was significantly down-regulated whereas miR-217 was obviously up-regulated in diabetic skin, HG-induced lymphatic endothelial cells (LECs) and diabetic mouse model. GAS5 sponged miR-217 to up-regulate Prox1. GAS5 overexpression or miR-217 inhibition rescued the impairments of cell viability, migration and lymphatic vessel formation and the facilitation of apoptosis of LECs caused by HG. Similar impacts were observed on the protein level of VEGFR-3, LYVE-1, and Prox1. GAS5 promoted wound healing and lymphangiogenesis in the diabetic mouse model. CONCLUSION: GAS5 sponged miR-217 to up-regulate Prox1 and promote lymphangiogenesis and diabetic wound healing. This might provide novel therapeutic strategy to improve the efficacy of diabetic wound healing.
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Diabetes Mellitus/metabolismo , Proteínas de Homeodominio/metabolismo , Linfangiogénesis , MicroARNs/metabolismo , ARN Largo no Codificante/metabolismo , Transducción de Señal , Proteínas Supresoras de Tumor/metabolismo , Cicatrización de Heridas , Animales , Línea Celular , Diabetes Mellitus/genética , Proteínas de Homeodominio/genética , Humanos , Ratones , MicroARNs/genética , ARN Largo no Codificante/genética , Proteínas Supresoras de Tumor/genéticaRESUMEN
Background: To reveal the alterations of tRNA-derived small RNA (tsRNA) expression profiles induced by hyperbaric oxygen (HBO) treatment in diabetic foot ulcers (DFUs) and investigate new therapeutic targets. Materials & methods: tsRNA sequencing was employed in normal skin tissue, in DFUs, and after HBO treatment groups. A quantitative real-time PCR was used to validate tsRNA sequencing results and their targets levels. Bioinformatics analysis was performed to reveal their therapeutic functions in DFUs. Results: A total of 22 tsRNAs were differentially expressed in the three groups. Three selected tsRNAs were validated by quantitative real-time PCR for further analysis, which were all significantly overexpressed in DFU while being normally expressed after HBO treatment. Bioinformatics analysis disclosed that these tsRNAs may play therapeutic roles through the regulation of the Wnt signaling pathway. Conclusion: tsRNAs may be novel useful targets for HBO to treat DFUs.
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Diabetes Mellitus , Pie Diabético , Oxigenoterapia Hiperbárica , Pie Diabético/genética , Pie Diabético/metabolismo , Pie Diabético/terapia , Humanos , Oxígeno , ARN de Transferencia/genética , Transducción de SeñalRESUMEN
BACKGROUND: The aim of this study is to identify the differentially expressed proteins in circulating polymorphonuclear neutrophils (PMN) from scalded bacteremia rabbits infected with Staphylococcus aureus to provide a basis to reveal the pathogenesis of burns and sepsis. METHODS: Rabbits were subjected to sham burn (A), A + bacterial challenge (B), 30% scald injury (C), or C + bacterial challenge (D). Bacterial challenge was inflicted as an injection of 2.0 x 10(8) cfu S. aureus 18 h after burn procedure. Animals were sacrificed 24 h after burn. PMN were isolated, and the differential proteins in the PMN from these animals were identified by two-dimensional electrophoresis coupled with MALDI-TOF-MS; two proteins were confirmed by Western blotting. RESULTS: Twenty-one differential protein spots were found, and seven differential proteins were identified. Among the identified proteins, the expression levels of protein disulfide-isomerase and thiol-specific antioxidant protein were down-regulated in groups C and D, and two protein spots of annexin I were identified, one of which was down-regulated and another up-regulated in groups C and D. CONCLUSIONS: Preliminary proteome changes in PMN from rabbits experiencing scald injury and S. aureus sepsis were revealed, which possibly play an important role in the inflammation and pathogenesis of sepsis after scald injury.