BACKGROUND:In previous studies,the equivalent structure of three-dimensional cell reconstruction of tissue engineering oral mucosa is similar to normal oral mucosa,including epithelial-like structure,lamina propria-like structure,and vascular lumen-like structure,and has initially achieved the establishment of vascular equivalent,but its vascularization characteristics are not very clear.OBJECTIVE:Vascular-like structures of vascularized oral mucosa equivalent were obtained by targeting vascular endothelial cells specific marker expression profiles correlated with laser capture microdissection system,and their vascularization ability was evaluated to reveal their vascularization characteristics.METHODS:Human gingival epithelial cells were cultured from human gingival epithelium and human gingival fibroblasts,human gingival mesenchymal stem cells were cultured from human gingival lamina propria.Human gingival mesenchymal stem cells were induced to differentiate into vascular endothelial-like cells after monoclonal expansion culture.Human gingival epithelial cells,human gingival fibroblasts,and vascular endothelial-like cells were loaded with acellular vascular matrix-0.25%human-like collagen type Ⅰ scaffold to construct the vascularized oral mucosa equivalent.The layered structure of oral mucosa equivalent(experimental group)and the acellular vascular matrix-0.25%human-like collagen type Ⅰ scaffold(control group)were implanted subcutaneously into the back of nude mice,respectively.14 days later,the incision surface of the two groups was coated with biogel.The biogel surface of the experimental group was inoculated with human gingival epithelial cells,while the control group was not inoculated with cells.The samples were collected after 14 days of feeding.The layered structure of oral mucosa equivalent was observed by morphology.The neovascular-like structures in oral mucosa equivalents were labeled by immunohistochemistry and immunofluorescence with a more comprehensive expression profile of vascular endothelial cells,and the vascularization characteristics were analyzed.A laser capture microdissection system was used to capture the neovascularization structures in the oral mucosa equivalents specifically labeled by immunohistochemistry and analyze their vascularization characteristics.RESULTS AND CONCLUSION:(1)The morphology showed that the cell level of oral mucosa equivalent was clear,and the structure was similar to that of normal oral mucosa,that is,there were epithelioid structures,lamina-like structures,and vascular cavelike structures,and there were scattered erythrocytes in the vascular cavelike structures.(2)The results of EdU Apollo tracer seed cells in the oral mucosa equivalent group showed that human gingival epithelial cells labeled with EdU Apollo 488 showed green fluorescence expression.DAPI labeled human gingival fibroblasts showed blue fluorescence expression and formed lamina-like structures in vivo.EdU Apollo 567 labeled vascular endothelial-like cells showed red fluorescence expression and formed a vascular-like structure in vivo.(3)Vascular endothelial cell specific marker expression profile immunofluorescence labeling of vascular structure showed that compared with normal oral mucosa,the expressions of CD31,CD51,CD54,CD105,Tie-2,VWF,vascular endothelial growth factor receptor 1,and vascular endothelial growth factor receptor 2 in oral mucosa equivalents were increased(P<0.000 1).There were no significant changes in CD34 expression(P>0.05).(4)Compared with the specifically labeled oral mucosal vascular structures,the expression levels of CD51,CD54,CD105,Tie-2,VWF,vascular endothelial growth factor receptor 1,and vascular endothelial growth factor receptor 2 of the oral mucosa equivalents targeted by the laser capture microdissection system were increased(P<0.000 1).There were no significant changes in expression of CD31 and CD34(P>0.05).(5)The results showed that the oral mucosa equivalent reconstructed by three-dimensional cell stratification could achieve good vascularization,and its vascularization characteristics were consistent with the immunological function and characteristics of neovascularization.Vascularization helps three-dimensional cell layer reconstruction of oral mucosa equivalent regeneration.

Objective To evaluate the predictive value of baseline peripheral blood immune cell profiles for immune-related adverse events(irAEs)following immune checkpoint inhibitors(ICIs)therapy for non-small cell lung cancer(NSCLC)patients.Methods In total,45 NSCLC patients who received ICIs treatment between December 2023 and December 2024 were enrolled.Patients were stratified into irAE(n=19)and non-irAE(n=26)groups based on irAE occurrence during treatment.Pre-treatment peripheral blood samples were collected,and flow cytometry was used to quantify immune subset proportions and activation marker expression.Receiver operating characteristic(ROC)curves were used to evaluate the predictive value of individual cellular markers for irAE development in NSCLC patients after immunotherapy.Results The irAE group exhibited significantly reduced proportion of intermediate monocytes(CD14+CD16+),as compared with the non-irAE group(P<0.05).The expression of CD69,an early activation marker,in CD3+T lymphocytes and the expression of CD154,an early activation marker,in CD8+T lymphocytes were both significantly lower in the irAE group than in the non-irAE group(P<0.05).ROC analysis demonstrated better predictive performance of the three-marker panel(intermediate monocytes/CD69+CD3+T cells and CD154+CD8+T cells;AUC=0.778)compared with individual biomarkers.Conclusion Lower levels of intermediate monocytes,CD69+CD3+T cells,and CD154+CD8+T cells in baseline peripheral blood may serve as potential biomarkers for predicting irAE development in NSCLC patients after immunotherapy.

Objective To evaluate the predictive value of baseline peripheral blood immune cell profiles for immune-related adverse events(irAEs)following immune checkpoint inhibitors(ICIs)therapy for non-small cell lung cancer(NSCLC)patients.Methods In total,45 NSCLC patients who received ICIs treatment between December 2023 and December 2024 were enrolled.Patients were stratified into irAE(n=19)and non-irAE(n=26)groups based on irAE occurrence during treatment.Pre-treatment peripheral blood samples were collected,and flow cytometry was used to quantify immune subset proportions and activation marker expression.Receiver operating characteristic(ROC)curves were used to evaluate the predictive value of individual cellular markers for irAE development in NSCLC patients after immunotherapy.Results The irAE group exhibited significantly reduced proportion of intermediate monocytes(CD14+CD16+),as compared with the non-irAE group(P<0.05).The expression of CD69,an early activation marker,in CD3+T lymphocytes and the expression of CD154,an early activation marker,in CD8+T lymphocytes were both significantly lower in the irAE group than in the non-irAE group(P<0.05).ROC analysis demonstrated better predictive performance of the three-marker panel(intermediate monocytes/CD69+CD3+T cells and CD154+CD8+T cells;AUC=0.778)compared with individual biomarkers.Conclusion Lower levels of intermediate monocytes,CD69+CD3+T cells,and CD154+CD8+T cells in baseline peripheral blood may serve as potential biomarkers for predicting irAE development in NSCLC patients after immunotherapy.

BACKGROUND:In previous studies,the equivalent structure of three-dimensional cell reconstruction of tissue engineering oral mucosa is similar to normal oral mucosa,including epithelial-like structure,lamina propria-like structure,and vascular lumen-like structure,and has initially achieved the establishment of vascular equivalent,but its vascularization characteristics are not very clear.OBJECTIVE:Vascular-like structures of vascularized oral mucosa equivalent were obtained by targeting vascular endothelial cells specific marker expression profiles correlated with laser capture microdissection system,and their vascularization ability was evaluated to reveal their vascularization characteristics.METHODS:Human gingival epithelial cells were cultured from human gingival epithelium and human gingival fibroblasts,human gingival mesenchymal stem cells were cultured from human gingival lamina propria.Human gingival mesenchymal stem cells were induced to differentiate into vascular endothelial-like cells after monoclonal expansion culture.Human gingival epithelial cells,human gingival fibroblasts,and vascular endothelial-like cells were loaded with acellular vascular matrix-0.25%human-like collagen type Ⅰ scaffold to construct the vascularized oral mucosa equivalent.The layered structure of oral mucosa equivalent(experimental group)and the acellular vascular matrix-0.25%human-like collagen type Ⅰ scaffold(control group)were implanted subcutaneously into the back of nude mice,respectively.14 days later,the incision surface of the two groups was coated with biogel.The biogel surface of the experimental group was inoculated with human gingival epithelial cells,while the control group was not inoculated with cells.The samples were collected after 14 days of feeding.The layered structure of oral mucosa equivalent was observed by morphology.The neovascular-like structures in oral mucosa equivalents were labeled by immunohistochemistry and immunofluorescence with a more comprehensive expression profile of vascular endothelial cells,and the vascularization characteristics were analyzed.A laser capture microdissection system was used to capture the neovascularization structures in the oral mucosa equivalents specifically labeled by immunohistochemistry and analyze their vascularization characteristics.RESULTS AND CONCLUSION:(1)The morphology showed that the cell level of oral mucosa equivalent was clear,and the structure was similar to that of normal oral mucosa,that is,there were epithelioid structures,lamina-like structures,and vascular cavelike structures,and there were scattered erythrocytes in the vascular cavelike structures.(2)The results of EdU Apollo tracer seed cells in the oral mucosa equivalent group showed that human gingival epithelial cells labeled with EdU Apollo 488 showed green fluorescence expression.DAPI labeled human gingival fibroblasts showed blue fluorescence expression and formed lamina-like structures in vivo.EdU Apollo 567 labeled vascular endothelial-like cells showed red fluorescence expression and formed a vascular-like structure in vivo.(3)Vascular endothelial cell specific marker expression profile immunofluorescence labeling of vascular structure showed that compared with normal oral mucosa,the expressions of CD31,CD51,CD54,CD105,Tie-2,VWF,vascular endothelial growth factor receptor 1,and vascular endothelial growth factor receptor 2 in oral mucosa equivalents were increased(P<0.000 1).There were no significant changes in CD34 expression(P>0.05).(4)Compared with the specifically labeled oral mucosal vascular structures,the expression levels of CD51,CD54,CD105,Tie-2,VWF,vascular endothelial growth factor receptor 1,and vascular endothelial growth factor receptor 2 of the oral mucosa equivalents targeted by the laser capture microdissection system were increased(P<0.000 1).There were no significant changes in expression of CD31 and CD34(P>0.05).(5)The results showed that the oral mucosa equivalent reconstructed by three-dimensional cell stratification could achieve good vascularization,and its vascularization characteristics were consistent with the immunological function and characteristics of neovascularization.Vascularization helps three-dimensional cell layer reconstruction of oral mucosa equivalent regeneration.

AIM:To establish a nomogram model to predict the effect of serum ferritin on diabetic retinopathy and evaluate the model.METHODS:A total of 21 variables, including ferritin, were screened by univariate and multivariate regression analysis to determine the risk factors of diabetic retinopathy. A nomogram prediction model was established for evaluation and calibration.RESULTS:Ferritin, duration of diabetes, hemoglobin, urine microalbumin, regularity of medication and body mass index were included in the nomogram model. The consistency index of the prediction model with serum ferritin was 0.813(95%CI: 0.748-0.879). The calibration curves of internal and external verification showed good performance, and the probability of the threshold suggested by the decision curve was in the range 10% to 90%. The model had a high net profit value.CONCLUSIONS:Serum ferritin is an important risk factor for diabetic retinopathy. A new nomogram model, which includes body mass index, duration of diabetes, ferritin, hemoglobin, urine microalbumin and regularity of medication, has a high predictive accuracy and could provide early prediction for clinicians.

AIM: To observe the clinical efficacy of 8-0 polypropylene scleral-sutured fixed intraocular lens(IOL)suspension implantation with the double knots technique in aphakic eyes.METHODS: Retrospective case series study. The data of 30 aphakic cases(31 eyes, 22 males)that underwent IOL suspension in our hospital from January 2021 to November 2022 were collected. The suspension of IOL(AcrySof IQ or Tecnis ZCB00)was performed by 8-0 polypropylene scleral-sutured with the double knots technique. The visual acuity, intraocular pressure(IOP), IOL position and complications with at least 6 mo of follow-up were observed.RESULTS: The mean preoperative uncorrected visual acuity(UCVA, LogMAR)and best-corrected visual acuity(BCVA, LogMAR)were 2.53±0.78 and 0.35±0.26, respectively, which were 0.58±0.26 and 0.36±0.27 at 6 mo postoperatively, respectively. And the differences in UCVA were statistically significant(t=15.408, P<0.01), whereas the difference in BCVA was not(t=-1.677, P=0.104). There were no intraoperative complications, with IOL position all centered, but 3 eyes had IOL tilt, 2 eyes had intraocular hypertension, 5 eyes had corneal edema, and 1 eye had suture exposure postoperatively. There were no complications such as hyphema, vitreous hemorrhage, macular edema, corneal endothelial decompensation, hypotony, choroidal detachment, retinal detachment, fulminant superior choroidal hemorrhage, endophthalmitis, or others.CONCLUSION: The 8-0 polypropylene scleral-sutured fixed intraocular lens suspension implantation with the double knots technique can improve the postoperative visual acuity of aphakic patients, and fewer complications, which is an option for the treatment of aphakia, dislocation of the lens and ligament abnormalities.

Arrhythmogenic cardiomyopathy (ACM), a fatal heart disease characterized by fibroadipocytic replacement of cardiac myocytes, accounts for 20% of sudden cardiac death and lacks effective treatment. It is often caused by mutations in desmosome proteins, with Desmoglein-2 (DSG2) mutations as a common etiology. However, the mechanism underlying the accumulation of fibrofatty in ACM remains unknown, which impedes the development of curative treatment. Here we investigated the fat accumulation and the underlying mechanism in a mouse model of ACM induced by cardiac-specific knockout of Dsg2 (CS-Dsg2 ^-/-). Heart failure and cardiac lipid accumulation were observed in CS-Dsg2 ^-/- mice. We demonstrated that these phenotypes were caused by decline of fatty acid (FA) β-oxidation resulted from impaired mammalian target of rapamycin (mTOR) signaling. Rapamycin worsened while overexpression of mTOR and 4EBP1 rescued the FA β-oxidation pathway in CS-Dsg2 ^-/- mice. Reactivation of PPARα by fenofibrate or AAV9-Pparα significantly alleviated the lipid accumulation and restored cardiac function. Our results suggest that impaired mTOR-4EBP1-PPARα-dependent FA β-oxidation contributes to myocardial lipid accumulation in ACM and PPARα may be a potential target for curative treatment of ACM.

BACKGROUND: In recent years, more and more scaffold materials have been used to repair soft tissue defects, and the clinical repair effect of soft tissue defects is strongly associated with the source and performance of materials. OBJECTIVE: To summarize the research progress of preparation and application of different biological scaffolds in the field of oral and maxillofacial soft tissue defect repair. METHODS: PubMed and Medline were searched for articles published from 1966 to 2019 with the English key words of “materials, scaffold, biological scaffold, soft tissue, coloboma, tissue engineering, review”. Chinese journal full-text database and Chinese science citation database were retrieved for articles published from 2003 to 2019 with key words of “material, scaffold, biological scaffold, soft tissue, defect, tissue engineering, review”. RESULTS AND CONCLUSION: Natural bio-scaffold materials are directly derived from organisms with pretty biocompatibility. Natural bio-scaffold materials can release cytoactive factors, promote cell adhesion, proliferation and differentiation, and can be combined with synthetic polymer materials with controllable properties to form composite scaffolds, which is an ideal scaffold material for repairing soft tissue defects of oral and maxillofacial regions. Nanomaterials have higher biological activity than other scaffold materials and can promote the adhesion and proliferation of seed cells, providing ideal three-dimensional space for cell growth, but their applications are currently mainly reflected in bone tissue repair, and the applications in soft tissue repair are very few. At present, there are many researches on natural biological scaffold materials in oral and maxillofacial soft tissue repair, mainly including small intestinal submucosa, acellular dermal matrix and acellular vascular scaffolds. The combination of natural biological scaffold materials and synthetic polymer materials will be a major research trend in materials for repairing oral and maxillofacial soft tissue defects.

Interleukin-27 (IL-27), a heterodimeric cytokine, plays a protective role in diabetes. Ghrelin, a gastric hormone, provides a hunger signal to the central nervous system to stimulate food intake. The relationship between IL-27 and ghrelin is still unexplored. Here we investigated that signal transducer and activator of transcription 3 (STAT3)-mechanistic target of rapamycin (mTOR) signaling mediates the suppression of ghrelin induced by IL-27. Co-localization of interleukin 27 receptor subunit alpha (WSX-1) and ghrelin was observed in mouse and human gastric mucosa. Intracerebroventricular injection of IL-27 markedly suppressed ghrelin synthesis and secretion while stimulating STAT3-mTOR signaling in both C57BL/6J mice and high-fat diet-induced-obese mice. IL-27 inhibited the production of ghrelin in mHypoE-N42 cells. Inhibition of mTOR activity induced by siRNA or rapamycin blocked the suppression of ghrelin production induced by IL-27 in mHypoE-N42 cells. siRNA also abolished the inhibitory effect of IL-27 on ghrelin. IL-27 increased the interaction between STAT3 and mTOR in mHypoE-N42 cells. In conclusion, IL-27 suppresses ghrelin production through the STAT3-mTOR dependent mechanism.