BACKGROUND:Previous studies have shown that 1,8-cineole has anti-inflammatory,antioxidation,antibacterial and anti-tumor effects.It has good anti-inflammatory effects in many diseases.OBJECTIVE:To investigate the effect of 1,8-cineole on inflammatory response in a rat model of experimental periodontitis.METHODS:Thirty Sprague-Dawley rats were assigned into normal control group,periodontitis control group and 1,8-cineole group with ten rats in each group according to the completely randomized digital table method.Except for the normal control group,rats in the other groups were induced into experimental periodontitis.The periodontitis model was constructed by the orthodontic ligature wire method.Eight weeks after modeling,in the 1,8-cineole group,1,8-cineole was placed into periodontal pockets,twice per day for 4 weeks.In the normal control group and the periodontitis control group,the same amount of normal saline was placed into periodontal pockets,twice per day for 4 weeks.After administration,general observation and periodontal clinical indicators were performed.Hematoxylin-eosin staining was used for periodontal histological evaluation.The expressions of inflammatory factors in the serum and gingiva at mRNA and protein levels were detected.RESULTS AND CONCLUSION:(1)Compared with the normal control group,rats in the periodontitis control group showed increased gingival bleeding index and periodontal probing depth(P<0.05),increased serum levels of interleukin 1β,tumor necrosis factor α,and interleukin 6(P<0.05),decreased serum level of interleukin 10(P<0.05),increased mRNA and protein levels of interleukin 1β,tumor necrosis factor α,and interleukin 6 in gingival tissue(P<0.05),and decreased mRNA and protein level of interleukin 10 in gingival tissue(P<0.05).Hematoxylin-eosin staining of periodontal tissues showed that compared with the normal control group,periodontal inflammation was obvious in the periodontitis control group.(2)Compared with the periodontitis control group,rats in the 1,8-cineole group showed decreased gingival bleeding index and periodontal probing depth(P<0.05),decreased serum levels of interleukin 1β,tumor necrosis factor α,and interleukin 6(P<0.05),increased serum level of interleukin 10(P<0.05),decreased mRNA and protein levels of interleukin 1β,tumor necrosis factor α,and interleukin 6 in gingival tissue(P<0.05),and increased mRNA and protein level of interleukin 10 in gingival tissue(P<0.05).Hematoxylin-eosin staining of periodontal tissues showed that compared with the periodontitis control group,periodontal inflammation was remarkably alleviated in the 1,8-cineole group.To conclude,1,8-cineole can attenuate the inflammatory response in the rat model of experimental periodontitis.

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 investigate the possibility of 1,8-cineole in the prevention of Streptococcus mutans(S.mutans)induced dental caries in rats.Methods:40 specific pathogen-free rats were randomly divided into 5 groups(n=8):distilled water group(negative control group),chlorhexidine group(positive control group),and eucalyptin low(MIC),medium(2MIC)and high(4MIC)dose groups.The plates were streaked with saliva samples,and calculated the number of S.mutans colonies and the ratio of the total number of colonies.Keyes' method was used to evaluate the caries damage of the jaws specimens.Results:At the end of the experiment,the proportion(％)of S.mutans in the oral cavity of the drug groups decreased(P＜0.000 1),but there was no significant difference between the drug group and the positive control group(P＜0.05).At the end of the experiment,compared with the negative control group,the scores of each grade in the drug group decreased(P＜0.05),but there was no significant differ-ence between positive control group and each 1,8-cineole group(P＞0.05),and among the 3 1,8-cineole dose groups(P＞0.05).Conclusion:1,8-cineole at the MIC can inhibit the growth of S.mutans in the oral cavity of rats with dental caries and reduce the frequency and severity of dental caries.

Objective To study how lipid bilayer fluidity modulates the interaction between β1 integrin and CD40L,as well as the formation of CD40L-mediated tumor cell contact interfaces.Methods Supported lipid bilayers(SLB)with different fluidities were prepared through adjusting the 1,2-dioleoyl-sn-glycero-3-[N-(5-amino-l-carboxypentyl)iminodiacetic acid]succinyl nickel salt(DGS-NTA)content.The functionalization of lipid bilayers was achieved by anchoring fluorescently labeled CD40L molecules onto the membrane surface.The contact interface formation of PC9 cells on the functionalized lipid bilayers was observed through confocal fluorescence imaging and fluorescence recovery after photobleaching(FRAP)experiments,and data of two dimensional(2D)reaction kinetics of β1 integrin and CD40L were extracted from Zhu-Golan plots.Results The diffusion coefficient of molecules in lipid bilayer was negatively correlated with DGS-NTA content.High fluidity of lipid bilayer promoted CD40L accumulation at cell contact interface and expanded the cell contact area.The 2D dissociation constants(2D Kd)of β1 integrin-CD40L complexes were approximately 13,31 and 65 molecules/μm2 for the three lipid bilayers with high,moderate and low fluidities,respectively.Conclusions High fluidity of lipid bilayers significantly facilitates diffusion and aggregation of CD40L to the cell contact interface,thus enhancing β1 integrin-CD40L interaction and the stability of cell contact interfaces.

Objective To study how lipid bilayer fluidity modulates the interaction between β1 integrin and CD40L,as well as the formation of CD40L-mediated tumor cell contact interfaces.Methods Supported lipid bilayers(SLB)with different fluidities were prepared through adjusting the 1,2-dioleoyl-sn-glycero-3-[N-(5-amino-l-carboxypentyl)iminodiacetic acid]succinyl nickel salt(DGS-NTA)content.The functionalization of lipid bilayers was achieved by anchoring fluorescently labeled CD40L molecules onto the membrane surface.The contact interface formation of PC9 cells on the functionalized lipid bilayers was observed through confocal fluorescence imaging and fluorescence recovery after photobleaching(FRAP)experiments,and data of two dimensional(2D)reaction kinetics of β1 integrin and CD40L were extracted from Zhu-Golan plots.Results The diffusion coefficient of molecules in lipid bilayer was negatively correlated with DGS-NTA content.High fluidity of lipid bilayer promoted CD40L accumulation at cell contact interface and expanded the cell contact area.The 2D dissociation constants(2D Kd)of β1 integrin-CD40L complexes were approximately 13,31 and 65 molecules/μm2 for the three lipid bilayers with high,moderate and low fluidities,respectively.Conclusions High fluidity of lipid bilayers significantly facilitates diffusion and aggregation of CD40L to the cell contact interface,thus enhancing β1 integrin-CD40L interaction and the stability of cell contact interfaces.

Objective:To investigate the possibility of 1,8-cineole in the prevention of Streptococcus mutans(S.mutans)induced dental caries in rats.Methods:40 specific pathogen-free rats were randomly divided into 5 groups(n=8):distilled water group(negative control group),chlorhexidine group(positive control group),and eucalyptin low(MIC),medium(2MIC)and high(4MIC)dose groups.The plates were streaked with saliva samples,and calculated the number of S.mutans colonies and the ratio of the total number of colonies.Keyes' method was used to evaluate the caries damage of the jaws specimens.Results:At the end of the experiment,the proportion(％)of S.mutans in the oral cavity of the drug groups decreased(P＜0.000 1),but there was no significant difference between the drug group and the positive control group(P＜0.05).At the end of the experiment,compared with the negative control group,the scores of each grade in the drug group decreased(P＜0.05),but there was no significant differ-ence between positive control group and each 1,8-cineole group(P＞0.05),and among the 3 1,8-cineole dose groups(P＞0.05).Conclusion:1,8-cineole at the MIC can inhibit the growth of S.mutans in the oral cavity of rats with dental caries and reduce the frequency and severity of dental caries.

BACKGROUND:Previous studies have shown that 1,8-cineole has anti-inflammatory,antioxidation,antibacterial and anti-tumor effects.It has good anti-inflammatory effects in many diseases.OBJECTIVE:To investigate the effect of 1,8-cineole on inflammatory response in a rat model of experimental periodontitis.METHODS:Thirty Sprague-Dawley rats were assigned into normal control group,periodontitis control group and 1,8-cineole group with ten rats in each group according to the completely randomized digital table method.Except for the normal control group,rats in the other groups were induced into experimental periodontitis.The periodontitis model was constructed by the orthodontic ligature wire method.Eight weeks after modeling,in the 1,8-cineole group,1,8-cineole was placed into periodontal pockets,twice per day for 4 weeks.In the normal control group and the periodontitis control group,the same amount of normal saline was placed into periodontal pockets,twice per day for 4 weeks.After administration,general observation and periodontal clinical indicators were performed.Hematoxylin-eosin staining was used for periodontal histological evaluation.The expressions of inflammatory factors in the serum and gingiva at mRNA and protein levels were detected.RESULTS AND CONCLUSION:(1)Compared with the normal control group,rats in the periodontitis control group showed increased gingival bleeding index and periodontal probing depth(P<0.05),increased serum levels of interleukin 1β,tumor necrosis factor α,and interleukin 6(P<0.05),decreased serum level of interleukin 10(P<0.05),increased mRNA and protein levels of interleukin 1β,tumor necrosis factor α,and interleukin 6 in gingival tissue(P<0.05),and decreased mRNA and protein level of interleukin 10 in gingival tissue(P<0.05).Hematoxylin-eosin staining of periodontal tissues showed that compared with the normal control group,periodontal inflammation was obvious in the periodontitis control group.(2)Compared with the periodontitis control group,rats in the 1,8-cineole group showed decreased gingival bleeding index and periodontal probing depth(P<0.05),decreased serum levels of interleukin 1β,tumor necrosis factor α,and interleukin 6(P<0.05),increased serum level of interleukin 10(P<0.05),decreased mRNA and protein levels of interleukin 1β,tumor necrosis factor α,and interleukin 6 in gingival tissue(P<0.05),and increased mRNA and protein level of interleukin 10 in gingival tissue(P<0.05).Hematoxylin-eosin staining of periodontal tissues showed that compared with the periodontitis control group,periodontal inflammation was remarkably alleviated in the 1,8-cineole group.To conclude,1,8-cineole can attenuate the inflammatory response in the rat model of experimental periodontitis.

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 severe pneumonia mouse model induced by bacterial infection.METHODS:A total of 102 male SPF C57BL/6J mice were randomly divided into a control group and a model group.Klebsiella pneu-moniae was administered via tracheal instillation at a concentration of 5×109 CFU.Mice were euthanized on days 1,2,4,8,and 14 post-infection to assess general condition,body weight,mortality,white blood cell and neutrophil counts,in-flammatory markers,and pathological changes in lung,heart,liver,spleen,kidney,and intestinal tissues.RESULTS:Mice in the model group exhibited symptoms such as dyspnea and huddling from 6 hours to 4 days post-infection,which progressively worsened,accompanied by continuous weight loss(P<0.01).These symptoms gradually resolved between days 5 and 14.Arterial oxygen saturation in the model group dropped to 80.7%from days 1 to 8(P<0.01)but returned to normal from days 9 to 14.A total of 23 model mice died between days 1 and 9,with no deaths thereafter,resulting in a mortality rate of 31.9%(P<0.01).Pathological examination revealed inflammatory cell infiltration,congestion,and ede-ma in lung tissue from days 1 to 2,with continued inflammatory cell infiltration,alveolar structural disorganization from days 4 to 8,and alveolar rupture and fusion by day 14(P<0.05 or P<0.01).Additionally,model mice showed significant increases in neutrophil count,white blood cell count,protein content in bronchoalveolar lavage fluid,total cell count,neutrophil ratio,and levels of inflammatory factors tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-6 in peripheral blood from days 1 to 8(P<0.05 or P<0.01).No significant pathological changes were observed in heart and liver tissues,while spleen,kidney,and intestinal tissues exhibited notable pathological changes:indistinct boundaries be-tween red and white pulp in the spleen,significant congestion and edema around renal glomeruli,renal tubules,and col-lecting ducts,and extensive inflammatory cell infiltration in the colonic mucosa.CONCLUSION:Tracheal instillation of 5×109 CFU Klebsiella pneumoniae induces severe pathological changes in the lungs of mice,offering a robust model for studying the pathogenesis and treatment of severe pneumonia.

The continuous strengthening of specialized capacity in hospitals has been gradually progressing in terms of discipline goal planning,talent development,technical cultivation,research and teaching collaboration,innovative management,and social services.Significant achievements have been made in this regard.Firstly,a preliminary talent development system has been established,forming a pyramid-shaped talent structure.Secondly,there has been a leap in the development of innovative technologies,with three medical new technologies recommended as provincial top ten medical new technologies in 2023.Thirdly,breakthroughs have been made in key specialized fields,with the hospital being approved for two national clinical key specialized construction projects and 39 provincial clinical key specialized projects.Fourthly,various indicators for high-quality development have steadily improved,with the hospital ranking 147th in the performance assessment of tertiary public hospitals in 2022 and be-ing the top-ranked municipal hospital in Hunan Province,maintaining an A-level grade for five consecutive years.