Current Status of Auricular Reconstruction Strategy Development.

Microtia has severe physical and psychological impacts on patients, and auricular reconstruction offers improved esthetics and function, alleviating psychological issues. Microtia is a congenital disease caused by a multifactorial interaction of environmental and genetic factors, with complex clinical manifestations. Classification assessment aids in determining treatment strategies. Auricular reconstruction is the primary treatment for severe microtia, focusing on the selection of auricular scaffold materials, the construction of auricular morphology, and skin and soft tissue scaffold coverage. Autologous rib cartilage and synthetic materials are both used as scaffold materials for auricular reconstruction, each with advantages and disadvantages. Methods for achieving skin and soft tissue scaffold coverage have been developed to include nonexpansion and expansion techniques. In recent years, the application of digital auxiliary technology such as finite element analysis has helped optimize surgical outcomes and reduce complications. Tissue-engineered cartilage scaffolds and 3-dimensional bioprinting technology have rapidly advanced in the field of ear reconstruction. This article discusses the prevalence and classification of microtia, the selection of auricular scaffolds, the evolution of surgical methods, and the current applications of digital auxiliary technology in ear reconstruction, with the aim of providing clinical physicians with a reference for individualized ear reconstruction surgery. The focus of this work is on the current applications and challenges of tissue engineering and 3-dimensional bioprinting technology in the field of ear reconstruction, as well as future prospects.

Gout-associated uric acid crystals induce tophi ulcerations and impair wound healing in a novel gouty ulcer model.

This study aimed to establish a novel gouty ulcer rat model induced by monosodium urate (MSU) deposition and preliminarily explored how MSU crystals affected wound healing. MSU crystals were subcutaneously injected into the back of rats to simulate tophi formation and ulceration. Ultrasound was used to detect the formation of gouty tophi. MSU crystal deposition and histopathological changes were analysed by haematoxylin-eosin staining. After the skin over the tophi became broken in the model group, a full-thickness tissue defect of the same area was made on the backs of the phosphate buffered saline (PBS) controls. On Days 3, 7, and 14 after wounding, the infiltration of neutrophils and macrophages and the expression of inflammatory markers, including interleukin-1ß (IL-1ß), tumour necrosis factor-α (TNF-α), and Nod-like receptor protein 3 (NLRP3), were examined by immunohistochemical staining and Western blotting, respectively. After the first subcutaneous injection in rats, local tissues showed redness and swelling, indicating inflammation on approximately Day 14. Tophi-like manifestations appeared on approximately Day 18. Tophi appeared heterogeneously hyperechoic by ultrasound. Swelling and redness in injured tissue areas increased on approximately Day 22, skin tissue necrosis was seen in a small area on approximately Day 26, and skin necrosis was enlarged and the tophi were ulcerated on approximately Day 32, accompanied by yellowish-white, chalky secretions. Haematoxylin and eosin staining showed dermal deposition of needle-like crystals with surrounding granulomatous inflammation. On Days 3, 7, and 14 after wounding, immunohistochemical staining showed the infiltration of neutrophils and macrophages, and the expression of inflammation-related proteins (IL-1ß, TNF-α, and NLRP3) were upregulated in gouty ulcers compared with those of PBS controls. The gouty ulcers were not completely healed by Day 14 compared with those in the PBS controls. In this study, a novel gouty ulcer rat model was constructed, which also revealed the existence of persistent chronic inflammation.

Foamy macrophages potentially inhibit tuberculous wound healing by inhibiting the TLRs/NF-κB signalling pathway.

To characterise the distribution, classification, and quantity of foamy macrophages (FMs) in tuberculous wound tissue and the relationship between FM and delayed healing of tuberculous wounds. Morphological studies were performed to explore the distribution of FM and Mycobacterium tuberculosis (Mtb) in tuberculous wounds, with acute and chronic wounds included for comparison. Phorbol-12-myristate-13-acetate stimulation-differentiated THP-1 cells were treated with Mtb to induce their differentiation into FM with oxidised low-density lipoprotein treatment serving as a control. Relative cytokine levels were determined by quantitative PCR and Western blotting. Varied co-culture combinations of Mtb, THP-1, FM, and fibroblasts were performed, and proliferation, migration, ability to contract collagen gel, and protein levels of the chemokines in the supernatants of the fibroblasts were assessed. The differentially expressed genes in human skin fibroblasts (HSFs) after co-culture with or without FM were identified using microarray. Many FM were found in the tissues of tuberculous wounds. The FM that did not engulf Mtb (NM-FM) were mainly distributed in tissues surrounding tuberculous wounds, whereas the FM that engulfed Mtb (M-FM) were dominantly located within granulomatous tissues. Co-culture experiments showed that, with the Mtb co-culture, the portions of NM-FM in the total FM grew over time. The migration, proliferation, chemokine secretion, and the ability of fibroblasts to contract collagen gel were inhibited when co-cultured with Mtb, FM, or a combination of the two. Further investigation showed that the TLRs/NF-κB signalling pathway is involved in fibroblast function under the stimulation of FM. TLRs and NF-κB agonists could reverse the phenotypic changes in HSFs after co-culture with FM. The tuberculous wound microenvironment composed of Mtb and FM may affect wound healing by inhibiting the functions of fibroblasts. FM potentially inhibit fibroblasts' function by inhibiting the TLRs/NF-κB signalling pathway in tuberculous wounds.

Current surgical practices of robotic-assisted tissue repair and reconstruction.

This paper systematically reviewed and analyzed the recent publications of robotic-assisted surgeries in the field of tissue repair and reconstruction. Surgical robots can elevate skin flap more accurately and shorten the time of tissue harvest. In addition, robotic-assisted surgery has the advantage of minimal tissue trauma and thus forms minimal scar. The utilization of surgical robots reduces the occurrence of complications after oral radical tumor resection while achieving cosmetic sutures. Robotic-assisted radical mastectomy could radically remove invasive breast cancer lesions and achieve breast reconstruction in the first stage through the small incisions in the operation areas. Surgical robots enable precise microvascular anastomosis and reduce tissue edema in the surgical field. Robotic-assisted technology can help appropriately locate the target tissues at different angles during sinus and skull base surgeries and accurately place tissues during urethroplasty. The robotic-assisted technology provides a new platform for surgical innovation in the field of tissue repair and reconstruction. However, the uncertainty in the survival rate after tumor radical surgery, the increase of operating time, and the high costs are barriers for its clinical application in tissue repair and reconstructive surgery. Nevertheless, robotic-assisted technology has already demonstrated an impact on the field of tissue repair and reconstruction in a meaningful way.

Characterization of Mycobacterium marinum infections in zebrafish wounds and sinus tracts.

The impaired healing of Mycobacterium tuberculosis-infected wounds is a clinical challenge, and the mechanisms involved are still not clear. The zebrafish model of Mycobacterium marinum infection has provided surprising insights into the pathogenesis of tuberculosis in humans. Similarly, the major principles and phases of cutaneous wound healing are conserved among adult mammals and adult zebrafish. Here, we injected Mycobacterium marinum into the dorsal muscles of adult zebrafish and observed the development of chronic wound pathology. Deep sequencing showed that gene expression related to muscles was down-regulated, whereas expressions of the IL-1ß, TNF-α, dram1 genes and the transcript of mir1-2 gene were up-regulated in infected wounds of zebrafish compared with control zebrafish. Muscles are immune-responsive tissues. Thus, muscles may play a role in the anti-Mycobacterium tuberculosis immunologic process, which leads to apoptosis of the infected muscle cell and formation of the subcutaneous sinus tract.

Three-Dimensional Printing of Reduction Template in the Contouring of Craniofacial Fibrous Dysplasia.

BACKGROUND: Contouring is a minimal invasive procedure to remove excess lesions and restore facial appearance of patients with craniofacial fibrous dysplasia; however, it is difficult to measure the amount of excess lesions. This study is to demonstrate the use of reduction template produced by three-dimensional printer in contouring procedure. METHODS: Computed tomography data were reconstructed into a three-dimensional model by Mimics software. The unaffected side was mirrored and superimposed on the affected side. Excess fibrous dysplasia lesions were calculated by Boolean operations. Lesions to be removed were separated and saved as stereolithography file. This model was further constructed in 3-matic package of Mimics software, and several holes were designed to indicate the variation in depth. Finally, the virtual model was exported into three-dimensional printer to produce physical template. RESULTS: With the direct guidance of three-dimensional reduction template, contouring procedure was performed easily and accurately. Good facial symmetry was achieved with reduced operative time and loss of bleeding. CONCLUSIONS: The application of reduction template produced by three-dimensional printer in contouring procedure improves the efficient and safety of surgery. Good outcomes are obtained without extra requirement in craniofacial surgery.

Ginsenoside rh2 inhibits cancer stem-like cells in skin squamous cell carcinoma.

BACKGROUND/AIMS: Treatments targeting cancer stem cells (CSCs) are most effective cancer therapy, whereas determination of CSCs is challenging. We have recently reported that Lgr5-positive cells are cancer stem cells (CSCs) in human skin squamous cell carcinoma (SCC). Ginsenoside Rh2 (GRh2) has been shown to significantly inhibit growth of some types of cancers, whereas its effects on the SCC have not been examined. METHODS: Here, we transduced human SCC cells with lentivirus carrying GFP reporter under Lgr5 promoter. The transduced SCC cells were treated with different doses of GRh2, and then analyzed cell viability by CCK-8 assay and MTT assay. The effects of GRh2 on Lgr5-positive CSCs were determined by fow cytometry and by tumor sphere formation. Autophagy-associated protein and ß-catenin were measured by Western blot. Expression of short hairpin small interfering RNA (shRNA) for Atg7 and ß-catenin were used to inhibit autophagy and ß-catenin signaling pathway, respectively, as loss-of-function experiments. RESULTS: We found that GRh2 dose-dependently reduced SCC viability, possibly through reduced the number of Lgr5-positive CSCs. GRh2 increased autophagy and reduced ß-catenin signaling in SCC cells. Inhibition of autophagy abolished the effects of GRh2 on ß-catenin and cell viability, while increasing ß-catenin abolished the effects of GRh2 on autophagy and cell viability. CONCLUSION: Taken together, our data suggest that GRh2 inhibited SCC growth, possibly through reduced the number of Lgr5-positive CSCs. This may be conducted through an interaction between autophagy and ß-catenin signaling.

[Effects of microRNA-155 on mice with lipopolysaccharide-induced acute lung injury].

OBJECTIVE: To study the effects of microRNA (miR)-155 on mice with acute lung injury induced by lipopolysaccharide (LPS). METHODS: According to the random number table, 30 C57 mice were divided into group LPS, group miR-155 inhibitor, group miR control, group miR-155 mimics, normal control group, with 6 mice in each group. Mice in group LPS and the other three treatment groups were given 5 µl LPS (20 µg/µl, in the dose of 4 mg/kg) to reproduce acute lung injury model. Half an hour after injury, group LPS, group miR-155 inhibitor, group miR control, group miR-155 mimics were injected with saline, miR-155 inhibitor, miR control and miR-155 mimics through trachea. Mice in normal control group were received no treatment. All mice were anesthetized after 24 h, while lung tissue samples were harvested for histomorphological observation, detection of mRNA expressions of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) with SYBR Green real-time polymerase chain reaction (PCR), detection of miR-155 in group miR-155 inhibitor, group miR control, group miR-155 mimics by Taqman PCR. The protein expression of cyclooxygenase-2 (COX-2) was determined by western blot. Data were statistically analyzed. RESULTS: The relative mRNA expression values of TNF-α were 24.49 ± 3.64, 38.92 ± 3.40, 36.00 ± 3.86, 12.56 ± 2.06, 1.04 ± 0.41 in group LPS, group miR-155 inhibitor, group miR control, group miR-155 mimics, normal control group. The relative mRNA expression values of IL-1ß were 55.96 ± 4.87, 62.90 ± 6.41, 59.99 ± 6.48, 25.33 ± 3.50, 1.11 ± 0.44 in group LPS, group miR-155 inhibitor, group miR control, group miR-155 mimics, normal control group. The differences of TNF-α and IL-1ß in group miR-155 mimics were statistically significant by compared with group LPS, group miR-155 inhibitor, group miR control (P < 0.05). The differences were not statistically significant among group LPS, group miR-155 inhibitor, group miR control. Western blot analysis showed the relative expression values of COX-2 were 0.94 ± 0.06, 0.96 ± 0.09, 0.91 ± 0.03, 0.79 ± 0.04, 0.63 ± 0.07 in group LPS, group miR-155 inhibitor, group miR control, miR-155 mimics and normal control group. The value in normal control group was significantly lower than other groups (P < 0.05). CONCLUSION: Using the miR-155 can effectively alleviate the lung inflammation in mice with acute lung injury induced by LPS.

Global burden of burns and its association with socio-economic development status, 1990-2019.

BACKGROUND: Burns represent important global health problems. Whereas many studies are limited by the difficulties in estimating the burden of burns and instead focus on the causes of burns, such as fire, heat, and hot substances. Therefore, a complete assessment of the burden of all injuries leading to burns is essential to developing reasonable global intervention strategies. METHODS: Data on three classes of burns, including "< 20 % total burned surface area without lower airway burns" (Moderate injury), "> =20 % total burned surface area or > = 10 % burned surface area if head/neck or hands/wrist involved w/o lower airway burns" (Major injury), "Lower airway burns" (Inhalation injury) were collected from the Global Burden of Disease 2019 database. Age-standardized incidence rates (ASR-I) and Years Lived with Disability (ASR-YLDs) for burns has been standardized by removing the influence of population size and age structure. They were extracted and stratified by cause, year, sex, age, socio-demographic index, country, and territory. RESULTS: In terms of ASR-I and ASR-YLDs, burns showed a significant decrease from 1990 to 2019, especially for moderate and major injury. In 2019, the burden of moderate injury was positively correlated with socio-demographic index while major injury was negatively correlated (P < 0.05). We found no correlation between socio-demographic index and the burden for inhalation injury (P > 0.05). Fire, heat, and hot substances were the most important cause of burns except for inhalation injury. The most common association with inhalation injury was falls, which were also a major cause of moderate and major injury. CONCLUSIONS: The Global Burden of Disease 2019 database data can be used to guide the allocation of resources to reduce ASR-I and ASR-YLDs of different burn classes.

Stem cell­mediated modulation of pyroptosis contributes to tissue repair in noninfective inflammatory­related diseases (Review).

Pyroptosis, a programmed cell death marked by lytic and inflammatory characteristics, plays a crucial role in non­infectious inflammation­related diseases but can lead to detrimental outcomes when dysregulated. Stem cells have emerged as key players in modulating pyroptosis through paracrine signaling, offering a novel avenue for tissue repair and regeneration. The present review delved into previous studies elucidating the intricate interplay between stem cells and pyroptosis, emphasizing the potential of stem cell­based therapies in regulating pyroptotic pathways. The exploration of this dynamic interaction holds promise for developing strategies to harness stem cells for effective tissue repair and regeneration in the context of inflammation­related diseases.

The ultra-early protective effect of ulinastatin on rabbit acute lung injury induced by paraquat.

OBJECTIVE: To study ultra-early pathophysiological changes of rabbit acute lung injury (ALI) caused by paraquat (PQ) and discuss the ultra-early protective effect of ulinastatin on rabbit ALI due to PQ. METHODS: 30 New Zealand white rabbits were randomly divided into a control group, a paraquat group and an ulinastatin intervention group with 10 rabbits in each group. For paraquat group and intervention group a single dose of paraquat (35 mg/kg) was injected intraperitoneally to establish rabbit models of ALI. The control group was injected an equal volume of saline. The intervention group was treated with 100 Ku/kg ulinastatin immediately after the establishment of the ALI model. The respective experimental groups underwent 320-slice CT perfusion scan of pleural at 2h, 4h and 6h time point after modeling to get CTP (CT Perfusion) images and related parameters. 2 mL blood was collected in the marginal ear vein to determine the mass concentration of the vascular endothelial growth factor (VEGF). The animals were killed by air embolism after 6h and lung tissue was taken for pathology observation. RESULTS: The reginal blood flow (rBF) and reginal blood volume (rBV) of paraquat group at 2,4,6 h time point were significantly (P <0.05) lower than those of control group. The intervention group rBF and rBV at 2, 4 and 6 h time points were significantly higher (P <0.05) compared to paraquat group. The permeability surface (rPS) and VEGF mass concentration of paraquat group at 2,4,6 h time point were significantly higher than the control group (P <0.05), and the intervention group rPS and VEGF mass concentrations at 2,4,6h time point were significantly lower (P <0.05) than those of paraquat group. Pathological detection indicators of paraquat group (congestive capillary percentage, the number of red blood cells outside of capillaries, percentage of capillaries with basement membrane damage) were significantly higher (P <0.05) at 6h time point compared with the control group, while significantly lower (P <0.05) in intervention group than in paraquat groups. Pathological observation under light microscope showed in paraquat group obvious inflammatory cell infiltration, alveolar epithelial cell hyperplasia, widened alveolar septum, visible focal hemorrhage, visible acute and chronic inflammatory cell infiltration in bronchioles cavity; under electron microscopy alveolar epithelial cell degeneration and necrosis, vascular welling of the endothelial cells, basement membrane rupture, a lot of exudates in alveolar space. In the intervention group, the above the symptoms were mitigated. CONCLUSION: In the ultra-early stage of rabbit ALI induced by PQ, pulmonary vascular endothelial cell is damaged and serum VEGF mass concentration and pulmonary vascular permeability increase. Early ulinastatin intervention can reduce serum VEGF level and PQ-induced vascular permeability amplitude, indicating that ulinastatin has a protective effect on pulmonary vascular endothelial cells.

[Application of expansion bilateral delto-pectoral thin skin flap in the repair of extensive scar in faciocervical region].

OBJECTIVE: To contrive an effective method of repairing the scar in bilateral faciocervical region. METHODS: Between April 2009 and February 2012, 9 patients with large scars on face and neck due to burn and scald were treated. There were 5 cases with face scars and 4 cervical scars. Their average age was 33 years (range: 23-48 years). The disease duration was 6 months to 20 years (mean: 6.5 years). The scar area was 12 cm × 7 cm to 22 cm × 26 cm. The soft tissue expanders (600-800 ml in volume) were implanted in delto-pectoral zone in one-stage operation. In two-stage operation, after the resection of cervical scars, the defects were repaired with delto-pectoral perforator flaps. In 5 facial scar cases, skin flap pedicle division was performed at Week 3. After the resection of scars, all wounds were repaired by expansion flap. The donor sites were sutured directly. The area of removed scar and the status of flap blood supply were observed. And the texture of flaps and patient satisfaction score were followed up for 6-30 months. RESULTS: Mild congestion of flap occurred postoperatively 1 case. The other flaps survived successfully. The flaps of 2 cases appeared bulky after transposition and flap repair was performed at Month 6. The appearance, texture, and color of flaps were similar to those at the donor sites. And there was an excellent match of flaps and recipient place.The patient satisfaction score was 7.6 ± 2.3. All achieved satisfactory functional and aesthetic outcomes. CONCLUSION: The method has many advantages and its clinical application is both safe and effective.

Transcriptomic analysis of mRNA expression in giant congenital melanocytic nevi.

BACKGROUND AND OBJECTIVE: GCMN is a sporadic disease with an incidence ranging from 1/20,000 to 1/500000. So far, several studies have found that GCMN is related to somatic mutations, but most of them have focused on known pathogenic genes, and transcriptome sequencing based on large datasets is relatively uncommon. At present, the use of next-generation sequencing technologies and bioinformatics platforms makes genomic information study more comprehensive and efficient. In this study, the transcriptome differences between GCMN lesions and surrounding normal skin tissues were investigated using high-throughput transcriptome sequencing, and hub genes and pathways related to pathogenesis were identified, providing a theoretical foundation for further research into the pathogenesis of GCMN. METHODS: Pathological skin tissue and surrounding normal skin tissue from GCMN patients, namely the pathological group (PG) and the control group (CG), were obtained. 1. All specimens were stained with HE to ensure that the samples met the experimental requirements. 2. Ten pairs of specimens were selected for high-throughput transcriptome sequencing, and the differentially expressed genes (DEGs) between the PG and the CG were obtained. The DEGs were analyzed by clusterProfiler R software for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The function of the subnetwork was analyzed and the hub genes were identified by the STRING database and Cytoscape software. 3. The expression differences of hub genes PTGS2, EGF, and SOX10 in pathological skin tissues and normal skin tissues were verified by qRT-PCR and immunofluorescence staining. RESULTS: 1. HE staining revealed a lot of melanocytes in the dermis and subcutaneous tissues. They were found around the hair follicles, sweat glands, sebaceous glands, and blood vessel walls, or in a specific pattern. 2. The screening threshold was set at p < 0.01 and |log2fc|<1, and a total of 1163 DEGs were discovered between the PG and CG, with 519 genes up-regulated and 644 genes down-regulated in the pathological tissues. According to the GO functional analysis, 29 biological processes, 18 cell compositions, and 17 molecular functions were significantly enriched, with the majority of them being related to keratinocytes and the extracellular matrix. There were 779 nodes and 2359 interactions in the protein interaction network. Using the MCODE plug-in, the network was divided into 25 functional clusters. According to ClueGO results, Cluster5 was involved in melanin biosynthesis and melanocyte proliferation. Using 11 operation methods in the Cytohubba plug-in, PTGS2, EGF, and SOX10 in Cluster5 were chosen as hub genes. 3. qRT-PCR and immunofluorescent staining revealed that compared to normal skin tissue, the expression of SOX10 was significantly up-regulated, and the expression of PTGS2 and EGF was significantly down-regulated in pathological skin tissue(P < 0.001). CONCLUSIONS: In GCMN, keratinocytes and extracellular matrix may directly and indirectly affect melanocyte activity. PTGS2, EGF, and SOX10 are important genes and significantly differentially expressed in pathological and normal skin tissues. These findings may serve as a springboard for future research.

Identifying miRNAs Associated with the Progression of Keloid through mRNA-miRNA Network Analysis and Validating the Targets of miR-29a-3p in Keloid Fibroblasts.

Background: Keloid has brought great trouble to people and currently has no uniformly successful treatment. It is urgent to find new targets to effectively prevent the progress of keloid. The current research mainly identifies the differentially expressed genes (DEGs) in keloid through high-throughput sequencing technology and bioinformatics analysis technology, to screen new therapeutic targets and potential biomarkers. However, due to the different samples, different control groups, and small sample sizes, the sequencing results obtained from different studies are quite different and lack reliability. It is necessary to analyze the existing datasets in a reasonable way. Methods: Datasets about keloid were filtered in Gene Expression Omnibus (GEO) and ArrayExpress databases according to the inclusion and exclusion criteria. The discovery datasets were used for summarizing significant DEGs, and the validation datasets were to validate the mRNA and miRNA expression levels. The Encyclopedia of RNA Interactomes (ENCORI) online platform was used to predict the interactions between miRNAs and their target mRNAs. Protein-protein interaction network (PPI network) analysis and functional enrichment analysis were conducted. miRNA-mRNA network was established by Cytoscape software and verified in keloid tissue (n = 8) by RT-qPCR. miR-29a-3p mimic and inhibitor were transfected into keloid fibroblasts (KFs) to preliminary verify its targets, the prognostic value of which was estimated by the receiver operating characteristic (ROC) curve. Results: A total of 6 datasets involving 20 patients were included. 15 miRNAs and 12 target mRNAs were identified as potential biomarkers for keloid patients. The RT-qPCR results showed that miR-29a-3p, miR-92a-3p, and miR-143-3p were downregulated, and all their target mRNAs were upregulated in keloid tissue (P < 0.05). The expression of COL1A1, COL1A2, COL3A1, COL5A1, and COL5A2 decreased when miR-29a-3p was overexpressed but increased when miR-29a-3p was knocked down (P < 0.05). And these genes had a good performance in the diagnosis of keloid, especially when using keloid nonlesional skin or normal scar tissues as controls. Conclusion: The miRNA-mRNA network, especially miR-29a-3p and its targets, may provide insights into the underlying pathogenesis of keloid and serve as potential biomarkers for keloid treatment.

Identification of Important Modules and Biomarkers That Are Related to Immune Infiltration Cells in Severe Burns Based on Weighted Gene Co-Expression Network Analysis.

Background: Immunosuppression is an important trigger for infection and a significant cause of death in patients with severe burns. Nevertheless, the prognostic value of immune-related genes remains unclear. This study aimed to identify the biomarkers related to immunosuppression in severe burns. Methods: The gene expression profile and clinical data of 185 burn and 75 healthy samples were obtained from the GEO database. Immune infiltration analysis and gene set variation analysis were utilized to identify the disorder of circulating immune cells. A weighted gene co-expression network analysis (WGCNA) was carried out to select immune-related gene modules. Enrichment analysis and protein-protein interaction (PPI) network were performed to select hub genes. Next, LASSO and logistic regression were utilized to construct the hazard regression model with a survival state. Finally, we investigated the correlation between high- and low-risk patients in total burn surface area (TBSA), age, and inhalation injury. Results: Gene set variation analysis (GSVA) and immune infiltration analysis showed that neutrophils increased and T cells decreased in severe burns. In WGCNA, four modular differently expressed in burns and controls were related to immune cells. Based on PPI and enrichment analysis, 210 immune-related genes were identified, mainly involved in T-cell inhibition and neutrophil activation. In LASSO and logistic regression, we screened out key genes, including LCK, SKAP1 and GZMB, and LY9. In the ROC analysis, the area under the curve (AUC) of key genes was 0.945, indicating that the key genes had excellent diagnostic value. Finally, we discovered that the key genes were related to T cells, and the regression model performed well when accompanied by TBSA and age. Conclusion: We identified LCK, SKAP1, GZMB, and LY9 as good prognostic biomarkers that may play a role in post-burn immunosuppression against T-cell dysfunction and as potential immunotherapeutic targets for transformed T-cell dysfunction.

LARP6 Regulates Keloid Fibroblast Proliferation, Invasion, and Ability to Synthesize Collagen.

Keloid is a skin fibroproliferative disease currently having no uniformly successful treatment. The lesion is composed of actively proliferating and collagen-overproducing fibroblasts. LARP6 is an RNA-binding protein able to regulate collagen synthesis in fibroblasts and to promote proliferation and invasion of tumor cells. To explore LARP6's likely functions in keloid pathogenesis, we performed immunohistochemistry staining on human keloid tissues and discovered markedly upregulated LARP6 expression in lesion fibroblasts compared with that of normal skin and hypertrophic scar tissues. In addition, the keloid tissue‒derived fibroblasts showed constitutive upregulation of LARP6 expression as well as significantly upregulated mRNA and protein expressions of type I collagen and enhanced cell proliferation and invasive behavior in cell culture system. Intriguingly, LARP6 knockdown by targeting with small interfering RNAs significantly inhibited type I collagen expression, proliferation, and invasion capability of keloid tissue‒derived fibroblasts relative to that of normal skin‒ and hypertrophic scar‒derived fibroblasts and control keloid tissue‒derived fibroblasts that were transfected with a scrambled small interfering RNA. In conclusion, the abnormally upregulated expression of LARP6 in fibroblasts may play an important role in the growth and invasive behavior of keloid lesions.

Identifying changes in immune cells and constructing prognostic models using immune-related genes in post-burn immunosuppression.

BACKGROUND: Burn patients are prone to infection as well as immunosuppression, which is a significant cause of death. Currently, there is a lack of prognostic biomarkers for immunosuppression in burn patients. This study was conducted to identify immune-related genes that are prognosis biomarkers in post-burn immunosuppression and potential targets for immunotherapy. METHODS: We downloaded the gene expression profiles and clinical data of 213 burn patients and 79 healthy samples from the Gene Expression Omnibus (GEO) database. Immune infiltration analysis was used to identify the proportion of circulating immune cells. Functional enrichment analyses were carried out to identify immune-related genes that were used to build miRNA-mRNA networks to screen key genes. Next, we carried out correlation analysis between immune cells and key genes that were then used to construct logistic regression models in GSE77791 and were validated in GSE19743. Finally, we determined the expression of key genes in burn patients using quantitative reverse transcription polymerase chain reaction (qRT-PCR). RESULTS: A total of 745 differently expressed genes were screened out: 299 were up-regulated and 446 were down-regulated. The number of Th-cells (CD4+) decreased while neutrophils increased in burn patients. The enrichment analysis showed that down-regulated genes were enriched in the T-cell activation pathway, while up-regulated genes were enriched in neutrophil activation response in burn patients. We screened out key genes (NFATC2, RORA, and CAMK4) that could be regulated by miRNA. The expression of key genes was related to the proportion of Th-cells (CD4+) and survival, and was an excellent predictor of prognosis in burns with an area under the curve (AUC) value of 0.945. Finally, we determined that NFATC2, RORA, and CAMK4 were down-regulated in burn patients. CONCLUSION: We found that NFATC2, RORA, and CAMK4 were likely prognostic biomarkers in post-burn immunosuppression and potential immunotherapeutic targets to convert Th-cell dysfunction.

Comparison of three different surgical approaches for the treatment of large/giant congenital melanocytic nevus.

OBJECTIVE: Treating large/giant congenital melanocytic nevus (L/GCMN) is challenging for surgeons. Operative approaches commonly used to remove L/GCMN include serial excision, tissue expansion, and skin grafting. Thus, we retrospectively compared these three operations' applications and therapeutic effects. METHODS: The clinical data of 97 L/GCMN patients from June 1, 2015, to June 1, 2019, were collected and divided into three groups according to the operations used: serial excision group (SE group, n = 18), tissue expansion group (TE group, n = 23), and skin grafting group (SG group, n = 56). The location and size of the lesion, the number of operations, duration of each operation, preoperative preparation time, postoperative hospital stay, complications, and clinical outcomes of all patients were collected and assessed. RESULTS: The SE group had the most times of operation (3.9 and 6.0 for LCMN and GCMN, respectively), the shortest surgery length (56.3 min), and the shortest postoperative hospital stay (10.0d). The SE and SG groups required much less time to prepare for surgery and had a lower rate of complications than the TE group. During the 11.9-month median follow-up period, the SE and TE groups had better postoperative outcomes than the SG group. CONCLUSION: Each of the three operations has different advantages and disadvantages, and the specific surgical strategy should be decided based on the patient's unique circumstances.

An improved explants culture method: Sustainable isolation of keloid fibroblasts with primary characteristics.

BACKGROUND: Keloid (KD) is a complex fibroproliferative disease, but the exact mechanisms underlying keloid pathogenesis remain to be elucidated. The primary keloid fibroblasts (KFs) culture in vitro has always been a fundamental measure to study the pathogenesis of keloid. However, the traditional primary culture methods have some limitations, such as a long culture cycle, low specimen utilization rate and so on. AIMS: Improve the keloid explants culture method sts. MATERIALS & METHODS: We proposed an improved new "explants multiple culture method"-reusing keloid explants for primary culture and harvesting the primary KFs in specific culture times. Meanwhile, the purity, proliferation, apoptosis, migration, invasion, extracellular matrix synthesis, and some fibrosis and inflammation-related proteins of KFs obtained from the first, fifth, and tenth explants cultures were detected. RESULTS: The results showed that the culture cycle of this new method (Cell Isolation: 2.67 ± 0.86 days, Explants removal: 8.83 ± 0.79 days, Cell Passage: 15.17 ± 1.39 days) was significantly shorter than that of the traditional method (Cell Isolation: 8.67 ± 1.84 days, Explants removal: 17.67 ± 2.17 days, Cell Passage: 22.67 ± 1.84 days). No significant difference was observed between the phenotypes of the fibroblasts obtained from the first explants culture and cultures less than 10 times (p > 0.05). DISSCUSSION: Taken together, this study provides an effective method for the primary culture of KFs with a higher specimen utilization rate and shorter culture cycle. CONCLUSION: This method breaks through the limitation of traditional explants culture requiring a large number of keloid specimens and provides a rich source of KFs for the study of keloid.

Satisfactory Wound Reconstruction with a Local Rotation Flap After Removal of Large Penile Divided Nevi: Original Technique, Early and Mid-Term Results.

Objective: To investigate the application of local rotation flaps for reconstruction of divided nevi of the penises in young male patients. Methods: A group of 8 patients of divided nevi of the penises who underwent wound reconstruction with local rotation flaps after surgical lesion removal was enrolled in a retrospective clinical study. Postoperative complication, sexual function and psychological traits were evaluated during the follow-up. Results: All patients, with ages ranged from 16 to 32 years (mean 23.25 years), were followed up for 6 to 48 months (mean 19.86 months). The patient's average length of hospital stay was 7.85 day (7 to 15 days). The average dimension of the lesions was (2.31±0.44) × (1.46±0.48) cm2 on the glans and (1.38±0.40) × (1.01±0.46) cm2 on the inner prepuce plate. All patients had no postoperative infection and were satisfied with the postoperative outcome upon discharge. Five cases of benign intradermal nevi and 3 cases of compound nevi without malignant transformation were confirmed by pathological evaluation on the removed samples. The sexual function of all patients was unaffected postoperatively by male sexual function scale (BMSFI and IIEF-5) evaluation. The psychological status of depression, anxiety and stress was all improved after the surgical reconstruction confirmed by the psychological traits scale (DASS) evaluation. Conclusion: Reconstruction with the local rotation flap is a simple, safe and appropriate surgical procedure, achieves satisfactory cosmetic outcome, and maintains intact male sexual function when used for the repair of defect after removal of divided nevi of the penises.