Purpose This study aimed to explore the mucinous phenotype characteristics,key points of differenti-al diagnosis and prognosis of invasive non-mucinous adenocarcinoma(INMA)and invasive mucinous adenocarcinoma(IMA)under the WHO(2021)lung adenocarcinoma classification.Methods We retrospectively collected clinico-pathological data from 522 cases of lung adenocarcinoma,including 425 INMA(66 with mucin secretion,259 without mucin secretion)and 97 IMA.Immunohistochemical(IHC)staining using the EnVision method was performed on the mucin-secreting adenocarcinoma to assess expression of TTF-1,HNF4α,MUC1,MUC4,MUC5AC,MUC5B,and MUC6.Unsupervised clustering analysis was conducted to explore phenotypic subgroups.Results 522 patients with lung adenocarcinoma ranged from 32 to 83 years old(median:61).251 cases(48.1％)were male and 271 cases(51.9％)were female.Clustering analysis divided lung adenocarcinomas into two major groups:one characterized by TTF-1-/HNF4α+and gastric-type mucins MUC5AC+/MUC6+,predominantly IMA;the other,TTF-1+/HNF4α-/MUC4+,largely INMA.A three-marker IHC panel(TTF-1,HNF4α,MUC6)distinguished IMA from mucinous IN-MA with an area under the ROC curve(AUC)of 0.957(95％CI:0.928-0.986)and a Youden's index of 0.860.Further cluster analysis of INMA cases identified four phenotypic subgroups.Prognostic analysis demonstrated that pa-tients with advanced-stage mucin-secreting INMA had significantly shorter overall survival(OS)and progression-free survival(PFS)than those without mucin secretion(5-year OS:57.1％ vs 81.8％,P=0.004;3-year PFS:40.9％ vs 62.4％,P=0.004).No significant survival differences were noted among INMA subgroups stratified by varying mucin proportions.Multivariate analysis identified pathological stage,tumor necrosis,KRAS mutation,and TTF-1 negativity as independent adverse prognostic factors for both OS and PFS in mucinous INMA.Conclusion A three-marker im-munohistochemical panel of TTF-1,HNF4α,and MUC6 is recommended to distinguish IMA from mucinous INMA.Mucus component portends a worse prognosis in advanced INMA,with necrosis,KRAS mutations,and TTF-1 negativi-ty serving as independent adverse prognostic factors in mucinous INMA.

Objective:To elucidate the regulatory role of matrix metallopeptidase 14 (MMP14) in the migration of chondrogenic precursor cells, thereby providing data support for investigating the pathogenesis of microtia.Methods:An in vitro differentiation model was established using human induced pluripotent stem cells (iPSCs) sequentially induced into neural crest cells (iNCCs) and subsequently into chondrogenic precursor cells (iCPCs), combined with lentivirus-mediated knockdown of MMP14, to investigate the effects of MMP14 on the biological characteristics of iCPCs, including proliferation, differentiation, and migration. Collective cell migration was assessed using scratch wound healing and Transwell migration assays; directional migration was characterized via high-content live-cell imaging; single-cell adhesion force was measured using a micromanipulation system. Collagen degradation was evaluated through hydroxyproline digestion assays. Cell proliferation was analyzed using the CCK-8 assay, and the expression of osteogenic/chondrogenic-related genes (SOX5/6/9, COL1A1, COL2A1, RUNX2, TWIST1) were quantified by real-time quantitative PCR. Immunofluorescence staining was used to assess the expression of F-actin and CD44 proteins. Additionally, transcriptomic sequencing was performed on iCPCs before and after MMP14 knockdown. Results:iPSC→iNCC→iCPC differentiation model was established in vitro. The resulting iCPCs expressed osteo/chondrogenic marker genes, including SOX5, SOX6, SOX9, COL1A1, COL2A1, RUNX2, and TWIST1, and exhibited positive expression of mesenchymal stem cell markers CD90, CD105, and CD73. Upon further induction, functional cartilage spheroids were formed. Compared with normal auricular chondrocytes, auricular chondrocytes from microtia patients showed reduced expression of MMP14 at both mRNA and protein levels. Lentivirus-mediated shRNA knockdown of MMP14 in iCPCs resulted in a marked decrease in its mRNA and protein expression. MMP14 knockdown significantly impaired collective migration of iCPCs, as evidenced by reduced wound closure rates in scratch assays and decreased numbers of migrated cells in Transwell assays. High-content live-cell imaging revealed that MMP14-deficient iCPCs displayed more erratic migration trajectories and a lower straight-line migration ratio. Single-cell adhesion assays showed extracellular matrix (ECM)-dependent alterations: cell adhesion was enhanced on matrigel-coated surfaces but weakened under uncoated conditions. MMP14 knockdown also led to reduced proliferation, decreased collagen degradation, diminished F-actin expression, fewer peripheral adhesion sites, and downregulation of CD44 protein expression, without significantly affecting the expression of chondrogenic genes such as SOX6, SOX9, COL1A1, COL2A1, RUNX2, and TWIST1. Transcriptomic analysis further revealed that MMP14 knockdown significantly downregulated genes involved in extracellular matrix organization, cell adhesion, migration, and tissue development, with enrichment in pathways including ECM-receptor interaction, focal adhesion, and MAPK signaling. Conclusion:MMP14 plays a critical role in the directional migration of chondrogenic precursor cells by regulating ECM remodeling, adhesion signaling, and cytoskeletal proteins.

Objective:To compare the physical properties of three-dimensional porous scaffolds with varying concentrations of acellular cartilage matrix (ACM), and to identify an optimal plan for the constructing tissue-engineered cartilage.Methods:(1) The cartilage particles were obtained by sampling, freezing, and crushing bovine knee cartilage. (2) ACM from was extracted from bovine articular cartilage through trypsin digestion, nuclease digestion, washing, and purification of cartilage particles. (3) Three-dimensional porous scaffolds were constructed using different concentrations of ACM: group A (100%), group B (60%), and group C (30%). (4) The scaffolds underwent gross observation, histological evaluation, DNA quantification, microstructural analysis, and measurement of physical properties.Results:Histological analysis revealed no discernible cell morphology or residual cell fragments in the ACM scaffolds across all groups. Hematoxylin and eosin (HE) staining, along with Sirius red staining, indicating the presence of collagen, suggesting retention to a certain extent. DNA quantification demonstrated a residual DNA concentration of (1.103 ± 0.174) ng/mg, confirming that the scaffolds were completely acellular. Scanning electron microscopy showed that the pore diameter of scaffolds in groups A, B, and C significantly decreased with the increase of ACM concentration, with no significant differences among the groups. Physical parameters indicated that scaffold density, yield strength, and Young’s modulus in groups A, B, and C were positively correlated with ACM concentration, while water absorption and porosity were negatively correlated. The pore diameters of scaffolds in all three groups ranged between 100 and 200 μm, with porosity exceeding 80%. Additionally, the water absorption and expansion rates were all above 450%, meeting the requirements for tissue engineering scaffolds. The Young’s modulus for scaffolds in group A and group B was (4.440±0.340) MPa and (0.963±0.320) MPa, respectively, which approached the mechanical properties of normal human articular cartilage.Conclusion:The three-dimensional porous scaffolds constructed with ACM at 100% and 60% concentrations demonstrate high water absorption and expansion rate, porosity, pore diameter, and mechanical properties comparable to those of normal cartilage tissue, making them suitable alternatives for tissue-engineered cartilage construction.

The chest deformity resulting from ear reconstruction using autologous costal cartilage significantly impacts patients’ physical development and overall quality of life. In recent years, numerous scholars have conducted extensive research on the diagnostic and therapeutic processes of chest deformities following costal cartilage surgery. The objective of this article is to comprehensively review and summarize the measurement indicators, mechanisms, influencing factors, prevention strategies, and research advancements pertaining to chest deformity following costal cartilage surgery. This will serve as a valuable reference for enhancing clinical practices.

Cartilage tissue engineering offers a novel therapeutic approach for auricular, tracheal, and joint reconstruction. Nevertheless, once implanted into the body, tissue-engineered cartilage is confronted with host immune rejection and inflammatory responses. These responses can result in cartilage deformation and fibrosis, thereby affecting the long-term maintenance of its morphology and function. Effectively modulating immune and inflammatory responses is crucial for advancing the clinical application of engineered cartilage. This article reviewed relevant literature to analyze the mechanisms of inflammatory responses occurring after the implantation of tissue-engineered cartilage, and summarized the types and surface characteristics of biomaterials, providing a reference for the design of biomaterials in cartilage tissue engineering.

Purpose This study aimed to explore the mucinous phenotype characteristics,key points of differenti-al diagnosis and prognosis of invasive non-mucinous adenocarcinoma(INMA)and invasive mucinous adenocarcinoma(IMA)under the WHO(2021)lung adenocarcinoma classification.Methods We retrospectively collected clinico-pathological data from 522 cases of lung adenocarcinoma,including 425 INMA(66 with mucin secretion,259 without mucin secretion)and 97 IMA.Immunohistochemical(IHC)staining using the EnVision method was performed on the mucin-secreting adenocarcinoma to assess expression of TTF-1,HNF4α,MUC1,MUC4,MUC5AC,MUC5B,and MUC6.Unsupervised clustering analysis was conducted to explore phenotypic subgroups.Results 522 patients with lung adenocarcinoma ranged from 32 to 83 years old(median:61).251 cases(48.1％)were male and 271 cases(51.9％)were female.Clustering analysis divided lung adenocarcinomas into two major groups:one characterized by TTF-1-/HNF4α+and gastric-type mucins MUC5AC+/MUC6+,predominantly IMA;the other,TTF-1+/HNF4α-/MUC4+,largely INMA.A three-marker IHC panel(TTF-1,HNF4α,MUC6)distinguished IMA from mucinous IN-MA with an area under the ROC curve(AUC)of 0.957(95％CI:0.928-0.986)and a Youden's index of 0.860.Further cluster analysis of INMA cases identified four phenotypic subgroups.Prognostic analysis demonstrated that pa-tients with advanced-stage mucin-secreting INMA had significantly shorter overall survival(OS)and progression-free survival(PFS)than those without mucin secretion(5-year OS:57.1％ vs 81.8％,P=0.004;3-year PFS:40.9％ vs 62.4％,P=0.004).No significant survival differences were noted among INMA subgroups stratified by varying mucin proportions.Multivariate analysis identified pathological stage,tumor necrosis,KRAS mutation,and TTF-1 negativity as independent adverse prognostic factors for both OS and PFS in mucinous INMA.Conclusion A three-marker im-munohistochemical panel of TTF-1,HNF4α,and MUC6 is recommended to distinguish IMA from mucinous INMA.Mucus component portends a worse prognosis in advanced INMA,with necrosis,KRAS mutations,and TTF-1 negativi-ty serving as independent adverse prognostic factors in mucinous INMA.

Objective:To elucidate the regulatory role of matrix metallopeptidase 14 (MMP14) in the migration of chondrogenic precursor cells, thereby providing data support for investigating the pathogenesis of microtia.Methods:An in vitro differentiation model was established using human induced pluripotent stem cells (iPSCs) sequentially induced into neural crest cells (iNCCs) and subsequently into chondrogenic precursor cells (iCPCs), combined with lentivirus-mediated knockdown of MMP14, to investigate the effects of MMP14 on the biological characteristics of iCPCs, including proliferation, differentiation, and migration. Collective cell migration was assessed using scratch wound healing and Transwell migration assays; directional migration was characterized via high-content live-cell imaging; single-cell adhesion force was measured using a micromanipulation system. Collagen degradation was evaluated through hydroxyproline digestion assays. Cell proliferation was analyzed using the CCK-8 assay, and the expression of osteogenic/chondrogenic-related genes (SOX5/6/9, COL1A1, COL2A1, RUNX2, TWIST1) were quantified by real-time quantitative PCR. Immunofluorescence staining was used to assess the expression of F-actin and CD44 proteins. Additionally, transcriptomic sequencing was performed on iCPCs before and after MMP14 knockdown. Results:iPSC→iNCC→iCPC differentiation model was established in vitro. The resulting iCPCs expressed osteo/chondrogenic marker genes, including SOX5, SOX6, SOX9, COL1A1, COL2A1, RUNX2, and TWIST1, and exhibited positive expression of mesenchymal stem cell markers CD90, CD105, and CD73. Upon further induction, functional cartilage spheroids were formed. Compared with normal auricular chondrocytes, auricular chondrocytes from microtia patients showed reduced expression of MMP14 at both mRNA and protein levels. Lentivirus-mediated shRNA knockdown of MMP14 in iCPCs resulted in a marked decrease in its mRNA and protein expression. MMP14 knockdown significantly impaired collective migration of iCPCs, as evidenced by reduced wound closure rates in scratch assays and decreased numbers of migrated cells in Transwell assays. High-content live-cell imaging revealed that MMP14-deficient iCPCs displayed more erratic migration trajectories and a lower straight-line migration ratio. Single-cell adhesion assays showed extracellular matrix (ECM)-dependent alterations: cell adhesion was enhanced on matrigel-coated surfaces but weakened under uncoated conditions. MMP14 knockdown also led to reduced proliferation, decreased collagen degradation, diminished F-actin expression, fewer peripheral adhesion sites, and downregulation of CD44 protein expression, without significantly affecting the expression of chondrogenic genes such as SOX6, SOX9, COL1A1, COL2A1, RUNX2, and TWIST1. Transcriptomic analysis further revealed that MMP14 knockdown significantly downregulated genes involved in extracellular matrix organization, cell adhesion, migration, and tissue development, with enrichment in pathways including ECM-receptor interaction, focal adhesion, and MAPK signaling. Conclusion:MMP14 plays a critical role in the directional migration of chondrogenic precursor cells by regulating ECM remodeling, adhesion signaling, and cytoskeletal proteins.

Objective:To compare the physical properties of three-dimensional porous scaffolds with varying concentrations of acellular cartilage matrix (ACM), and to identify an optimal plan for the constructing tissue-engineered cartilage.Methods:(1) The cartilage particles were obtained by sampling, freezing, and crushing bovine knee cartilage. (2) ACM from was extracted from bovine articular cartilage through trypsin digestion, nuclease digestion, washing, and purification of cartilage particles. (3) Three-dimensional porous scaffolds were constructed using different concentrations of ACM: group A (100%), group B (60%), and group C (30%). (4) The scaffolds underwent gross observation, histological evaluation, DNA quantification, microstructural analysis, and measurement of physical properties.Results:Histological analysis revealed no discernible cell morphology or residual cell fragments in the ACM scaffolds across all groups. Hematoxylin and eosin (HE) staining, along with Sirius red staining, indicating the presence of collagen, suggesting retention to a certain extent. DNA quantification demonstrated a residual DNA concentration of (1.103 ± 0.174) ng/mg, confirming that the scaffolds were completely acellular. Scanning electron microscopy showed that the pore diameter of scaffolds in groups A, B, and C significantly decreased with the increase of ACM concentration, with no significant differences among the groups. Physical parameters indicated that scaffold density, yield strength, and Young’s modulus in groups A, B, and C were positively correlated with ACM concentration, while water absorption and porosity were negatively correlated. The pore diameters of scaffolds in all three groups ranged between 100 and 200 μm, with porosity exceeding 80%. Additionally, the water absorption and expansion rates were all above 450%, meeting the requirements for tissue engineering scaffolds. The Young’s modulus for scaffolds in group A and group B was (4.440±0.340) MPa and (0.963±0.320) MPa, respectively, which approached the mechanical properties of normal human articular cartilage.Conclusion:The three-dimensional porous scaffolds constructed with ACM at 100% and 60% concentrations demonstrate high water absorption and expansion rate, porosity, pore diameter, and mechanical properties comparable to those of normal cartilage tissue, making them suitable alternatives for tissue-engineered cartilage construction.

Based on the principle of selective photothermodynamics, specific laser can be selectively absorbed by melanin particles in the hair follicles, resulting in a photothermal effect, through which hairball and hair follicle stem cells are completely destroyed, rendering permanent hair removal. Laser and light technology have the characteristics of convenience, efficiency, safety and permanent effect. Since its application in the reconstruction of the outer ear, it has helped solve the problem of residual hair and improve the shape of the auricle and the satisfaction of patients. The authors reviewed the laser types, treatment parameters, timing and interval, as well as related complications and corresponding treatment measures.

Cartilage tissue engineering offers a promising therapeutic approach for auricular, tracheal, and joint reconstruction. However, implanted engineered cartilage currently faces challenges such as host immune rejection and inflammatory reactions, which can lead to deformities and fibrosis, compromising long-term maintenance of shape and function. Effectively modulating immune and inflammatory responses is crucial for advancing the clinical application of engineered cartilage. This article reviewed relevant literature to analyze the mechanisms of inflammatory responses occurring after the implantation of tissue-engineered cartilage, and summarized the types and surface characteristics of biomaterials, providing a reference for the design of biomaterials in cartilage tissue engineering.