Smoking is a well-established risk factor for periodontitis, yet the precise mechanisms by which smoking contributes to periodontal disease remain poorly understood. Recent advances in spatial transcriptomics have enabled a deeper exploration of the periodontal tissue microenvironment at single-cell resolution, offering new opportunities to investigate these mechanisms. In this study, we utilized Visium HD single-cell spatial transcriptomics to profile gingival tissues from 12 individuals, including those with periodontitis, those with smoking-associated periodontitis, and healthy controls. Our analysis revealed that smoking disrupts the epithelial barrier integrity, induces fibroblast alterations, and dysregulates fibroblast-epithelial cell communication, thereby exacerbating periodontitis. The spatial analysis showed that endothelial cells and macrophages are in close proximity and interact, which further promotes the progression of smoking-induced periodontal disease. Importantly, we found that targeting the endothelial CXCL12 signalling pathway in smoking-associated periodontitis reduced the proinflammatory macrophage phenotype, alleviated epithelial inflammation, and reduced alveolar bone resorption. These findings provide novel insights into the pathogenesis of smoking-associated periodontitis and highlight the potential of targeting the endothelial-macrophage interaction as a therapeutic strategy. Furthermore, this study establishes an essential information resource for investigating the effects of smoking on periodontitis, providing a foundation for future research and therapeutic development for this prevalent and debilitating disease.
Humans ; Gingiva/cytology* ; Smoking/adverse effects* ; Male ; Periodontitis/pathology* ; Single-Cell Analysis ; Female ; Adult ; Middle Aged ; Macrophages ; Fibroblasts ; Endothelial Cells ; Case-Control Studies ; Chemokine CXCL12/metabolism*

Objective:To investigate the regulatory effect and mechanism of interleukin-22 (IL-22) on the gingival epithelial barrier in the context of periodontal inflammation.Methods:IL-22 knockout (IL-22 KO) mice were constructed, and periodontitis mice models were established through oral gavage with polymicrobial inoculation. DNAs were extracted from the oral plaques of IL-22 KO periodontitis mice group ( n=7) and their wild-type littermates periodontitis group ( n=7) to establish a periodontitis-related oral microbiota database"PD-RiskMicroDB", determining the relationship between changes in oral microbiota and microbial function in two groups using 16S rRNA sequencing results. Gingival epithelial cells (GEC) were cultured by modified trypsinization method, and were stimulated with 100 μg/L IL-22, Porphyromonas gingivalis (Pg) (multiplicity of infection:100), separately or together for 3 and 12 hours. The experimental groups were as follows: control group (no stimulation), IL-22 group, Pg group and Pg+IL-22 group. The expression of barrier protein E-cadherin in each group at 3 h was detected by immunofluorescence, real-time fluorescence quantitative PCR (RT-qPCR) and Western blotting. Fluorescein isothiocyanate-dextran-mediated epithelial cell permeability experiment was conducted to clarify the changes in permeability of GEC in each group at 3 and 12 h. The mRNA expressions of E-cadherin in the gingival epithelium of wild-type littermates periodontitis group and IL-22 KO periodontitis group were detected by RT-qPCR. Fifteen C57BL/6 wild-type mice were randomly divided into control group ( n=5), periodontitis group ( n=5) and periodontitis+IL-22 treatment group ( n=5). RT-qPCR and immunohistochemistry (IHC) staining were used to detect the expression level of E-cadherin in the gingival epithelium of each group. Results:16S rRNA sequencing results showed that the composition of oral microbiota changed in IL-22 KO periodontitis group, of which the abundance of bacterial genera related to periodontal tissue invasion was significantly increased (linear discriminant analysis score: 2.22, P=0.009), compared with wild-type littermates periodontitis group. In vitro cell experiments showed that after Pg infection for 3 hours, the cell connections of GEC in Pg group were interrupted, and the fluorescence intensity of E-cadherin was reduced in Pg group compared with the control group. Meanwhile, the mRNA and protein expression levels of E-cadherin (mRNA: 0.69±0.12; protein: 0.60±0.12) were downregulated compared with the control group [mRNA: 1.00±0.00 ( P=0.043); protein: 1.04±0.08 ( P=0.003)], respectively. The fluorescence intensity of E-cadherin in the Pg+IL-22 group was enhanced compared with Pg group, and expression levels of E-cadherin mRNA (1.16±0.10) and protein (0.98±0.07) in Pg+IL-22 group showed a significant increase compared with Pg group [mRNA: 0.69±0.12 ( P=0.005); protein: 0.60±0.12 ( P=0.007)]. The result of epithelial permeability test showed that there was no statistical difference in epithelial permeability among control group, Pg group, IL-22 group and Pg+IL-22 group with treatment for 3 hours ( F=0.20, P=0.893). While when the treatment time turned to be 12 hours, the epithelial barrier permeability showed a significant increase in Pg group (1.39±0.15) compared with control group (1.00±0.00, P=0.027), and a decrease in Pg+IL-22 group (1.02±0.18) compared with Pg group (1.39±0.15, P=0.034). In vivo, the mRNA expression of E-cadherin in the gingival epithelium of IL-22 KO periodontitis group decreased significantly (0.32±0.21) compared with wild-type littermates periodontitis group (1.01±0.01) ( t=5.70, P=0.005). Moreover, RT-qPCR and IHC staining results showed that the mRNA expression level of E-cadherin (0.40±0.07) and absorbance value of E-cadherin positive expression (0.02±0.00) in gingival epithelial tissue of periodontitis group were both significantly down-regulated compared with control group [mRNA: 1.00±0.00 ( P=0.005); absorbance value of E-cadherin positive expression: 0.04±0.01 ( P=0.006)]. Meanwhile, the mRNA expression level of E-cadherin (1.06±0.24) and the absorbance value of E-cadherin positive expression (0.03±0.01) were both observed increase in periodontitis+IL-22 treatment group compared with periodontitis group ( P=0.003, P=0.039). Conclusions:IL-22 may exert a protective effect on the gingival epithelial barrier in an inflammatory environment by regulating the invasiveness of oral microbiota and the expression of host barrier protein.

Objective:To investigate the relationship between C3, C4, Th1/Th2 levels and the Myasthenia Gravis Daily Living Scale (MG-ADL) score in patients with myasthenia gravis (MG) and its efficacy in predicting the transition of ocular muscle type to systemic type.Methods:A retrospective study of 94 patients with ophthalmic MG admitted to Haikou People's Hospital from April 2017 to April 2020 was conducted. According to whether they had converted to systemic MG within 6 months, they were divided into transformation group ( n=35) and non-transformation group ( n=59). The levels of C3, C4 and Th1/Th2, as well as the score of MG-ADL and Quantitative Myasthenia Gravis (QMG) were compared between the two groups before and 1 and 3 months after treatment. The correlation between C3, C4 and Th1/Th2 levels and MG-ADL and OMG scores, as well as the related influencing factors of the transformation from ocular muscle type to systemic type was analyzed. The efficiency of each index in predicting the transformation from ocular muscle type to systemic type was analyzed. Results:At 1 and 3 months after treatment, the C3 and C4 in both groups were significantly higher than before treatment, and Th1/Th2 was significantly lower than before treatment; the C3 and C4 in the non-transformation group were higher than that in the transformation group, while Th1/Th2 was lower than that in the transformation group (all P<0.05). The MG-ADL and QMG scores in 2 groups at 1 and 3 months after treatment were significantly lower than those before treatment, and those in the non-transformation group were lower than those in the transformation group (all P<0.05). C3 and C4 levels were negatively correlated with MG-ADL and QMG scores (all P<0.05), while Th1/Th2 levels were positively correlated with MG-ADL and QMG scores (all P<0.05). At 1 and 3 months after treatment, C3, C4 and Th1/Th2 were the influencing factors for the transformation from ocular muscle type to systemic type (all P<0.05). The area under the curve (AUC) of C3, C4 and Th1/Th2 combined to predict the transformation from ocular muscle type to systemic type at 3 months after treatment was 0.939, and the best predictive sensitivity and specificity were 91.43% and 88.14%, respectively. Conclusions:There is a good linear relationship between C3, C4, Th1/Th2 levels and MG-ADL scores in MG patients, and it has a high efficiency in predicting the transition of ocular muscle type to systemic type.

Objective:To investigate the correlation between serum neutrophil to lymphocyte ratio (NLR), thrombospondin 1 (TSP-1), miR-210 and systemic lupus erythematosus disease activity index (SLEDAI) and the prognostic value of their combination.Methods:The medical records of 126 patients with systemic lupus erythematosus (SLE) in Haikou People′s Hospital (Haikou Hospital Affiliated to Xiangya Medical College of Central South University) from February 2018 to February 2020 were retrospectively analyzed. According to the recurrence of prognosis 6 months after treatment, they were divided into recurrence group ( n=23) and non recurrence group ( n=103). The general data, serum NLR, TSP-1, miR-210 levels and SLEDAI score before and after treatment of the two groups were compared. The relationship between the levels of serum indicators before and after treatment, SLEDAI score, prognosis and recurrence of SLE patients were analyzed, and the efficacy of single and combined serum indicators in predicting prognosis was explored. Results:The levels of serum NLR, TSP-1, miR-210 and SLEDAI score in the recurrence group were higher than those in the non recurrence group before and after treatment (all P<0.05); After treatment, the levels of serum NLR, TSP-1, miR-210 and SLEDAI score in the two groups were lower than those before treatment (all P<0.05). Pearson correlation analysis showed that the levels of serum NLR, TSP-1 and miR-210 in SLE patients were positively correlated with SLEDAI scores (all P<0.05); Cox regression analysis showed that after adjusting other factors such as complement C3, complement C4 levels and SLEDAI scores before and after treatment, serum NLR, TSP-1 and miR-210 before and after treatment were still significantly correlated with the risk of recurrence in SLE patients (all P<0.05); The receiver operating characteristic (ROC) curve showed that the area under curve (AUC) of serum NLR, TSP-1 and miR-210 combined to predict recurrence was 0.907 (95% CI: 0.842-0.951), the sensitivity was 86.96%, and the specificity was 83.50%, which was significantly higher than that of each index alone. Conclusions:Serum NLR, TSP-1, miR-210 levels in SLE patients are positively correlated with SLEDAI scores, and the combined detection of these indicators has a high predictive value for prognosis and recurrence, which can provide references for the diagnosis and treatment of SLE.

Objective To study the diagnostic value of non-contrast T1mapping in left ventricular hypertrophy(LVH).Methods Forty LVH patients(LVH group)including 11 cardiac amyloidosis(CA),19 hypertrophic cardiomyopathy (HCM) and 10 hypertensive heart disease (HHD) patients, and 14 healthy volunteers (control group) were enrolled in this retrospective study between November 2015 and October 2016.All subjects underwent cardiac magnetic resonance(CMR)on a 3 T scanner.The CMR scan protocol included cine sequences, first-pass perfusion, late Gadolinium enhancement (LGE) and non-contrast T1 mapping(MOLLI)prototype sequences.The cardiac morphology was assessed by cine,first-pass perfusion as well as LGE.Left-ventricular end-diastolic wall thickness(EDTH)was assessed for 16 segments,native T1 values were measured in hypertrophic segments. The differences in EDTH and native T1values between LVH group and control group were evaluated using t test. The ANOVA and LSD were used in the comparison of differences among four sub-groups.Sensitivity,specificity,cut-off values and area under the curve (AUC) were derived using receiver-operating characteristics curve (ROC) analysis. Results The EDTH and native T1values in LVH group were significantly higher than those of control group[(16.5±5.2)mm vs.(6.3±1.8)mm,(1 388.6±119.8)ms vs.(1 248.4±58.1)ms,t=28.8 16.4,both P<0.01].Moreover,CA showed significantly higher T1value [(1 495.5 ± 100.9)ms] than that of HCM [(1 342.0 ± 69.2)ms] and HDD [(1 290.7±45.5)ms](F=300.5,P<0.01),and T1values in HCM were also higher than HDD(P<0.01).HCM showed significantly higher EDTH than that of CA and HDD (P<0.01), and EDTH in CA was also higher than HDD (P<0.01). The native T1showed good diagnostic performance between CA and HCM with AUC 0.914,sensitivity 90.1%%,and specificity 84.3%,and cutoff value 1 382.8 ms,between CA and HHD with AUC 0.989,sensitivity 97.0%,specificity 93.5% and cutoff value 1 359.5 ms.Conclusion The elevated native T1values were useful for quantitatively differential diagnosis of LVH.

Objective To investigate the influence of long non-coding RNA HOTAIR in proliferation and apoptosis of human tongue squamous cell carcinoma in vitro and in vivo. Methods siHOTAIR was used to inhibit the HOTAIR expression in Tb3.1 human tongue squamous cell carcinoma cell line. The experiments were divided into siHOTAIR group, nonsense sequence group and blank control group. Real-time PCR was used to detect the HOTAIR expression. MTT assay was employed to determine the cell survival. The expression levels of Bcl2, BAX, caspase-3, cleaved caspase-3 were examined by Western blot assay. Tb3.1 xenograft tumor model was established in BALB/c nude mice, and the tumor model was divided into control group, negative group, and siHOTAIR treated group. The tumor tissues were measured by immunohistochemistry stain (IHC) and TUNEL assay. Results The detection of real-time PCR showed that HOTAIR expression was reduced after treated with siHOTAIR. Western blots assay showed that Bcl-2 protein was suppressed while cleaved caspase-3 and BAX protein were up-regulated after treated with siHOTAIR. MTT assay indicated that the cell survival rate was significantly reduced in siHOTAIR treated group. Flow cytometry detected that apoptosis levels were increased in siHOTAIR group. The level of cell senescence was higher in the siHOTAIR group than that of control group. Results of IHC indicated that Ki-67 and Bcl-2 protein of tumor tissue were inhibited, while BAX and cleaved caspase-3protein expressions were elevated simultaneously in the siHOTAIR group. TUNEL assay suggested that more apoptosis was observed in siHOTAIR group. Conclusion HOTAIR can affect proliferation and apoptosis of tongue squamous cancer cells. HOTAIR may be one of the new candidate targets for human tongue cancer therapy.

Currently, a growing number of community-based organizations are providing rapid HIV testing service in various forms, some people with specific needs also purchase HIV rapid test papers through online sales channels, those imply that the demand of HIV self-test is in increasing year by year.In this paper, aims to understand the current situation of HIV rapid test led by CBOs and the approach, strategies and results of social marketing by means of expert interviews and site visits. Hope to illustrate the current situation, and make recommendations for future work.

Currently, a growing number of community-based organizations are providing rapid HIV testing service in various forms, some people with specific needs also purchase HIV rapid test papers through online sales channels, those imply that the demand of HIV self-test is in increasing year by year.In this paper, aims to understand the current situation of HIV rapid test led by CBOs and the approach, strategies and results of social marketing by means of expert interviews and site visits. Hope to illustrate the current situation, and make recommendations for future work.

Objective To explore the expressions of Cyclin-dependent kinase 5 (CDK5) and Epithelial-Mesenchymal Transition (EMT) related proteins including N-cadherin, Vimentin and E-cadherin in head and neck squamous cell carcino? ma (HNSCC), and to determine the relationship between the expression of CDK5 and prognosis. Methods The expression levels of CDK5 and EMT related proteins were evaluated by immunohistochemistry in 55 patients who were diagnosed as HN?SCC. They were also analyzed in different clinical pathological factors. The correlation of CDK5 and EMT related proteins as well as the relationship between the expression of CDK5 and prognosis were also analyzed. Results The expression level of CDK5 was significantly higher in patients with lymph node metastasis than that in patients with non-lymph node metastasis (91.67%vs 30.23%, P<0.05). It’s also higher in T3-T4 stages than that in T1-T2 stages (85%vs 20%, P<0.05). The ex?pression levels of N-cadherin and Vimentin were significantly higher in patients with lymph node metastasis than those in patients with non-lymph node metastasis (75.00%vs 6.98%;91.67%vs 27.91%, all P<0.05). However, the expression level of E-cadherin was significantly lower in patients with lymph node metastasis (8.33%vs 86.05%, P<0.05) compared to that in patients without. CDK5 was positively correlated with N-cadherin and Vimentin, but negatively correlated with E-cad?herin (rs=0.512, 0.443,-0.363, all P<0.01). The 3-year survival rates were significantly lower in patients with high expres?sion of CDK5 (37.5%) than that in patients with low expression of CDK5 (87%, Log-rankχ2=12.678, P<0.01). Conclusion CDK5 and EMT related proteins were activated abnormally in HNSCC with lymph node metastasis. CDK5 may be a new bio?logical marker for prognosis of HNSCC.

To sum up the reform of internal performance appraisal system and income allocation system of Shanghai municipal hospitals.The internal performance appraisal index system,evaluation methods and corresponding income allocation system,featuring two breaks,one change and eight elements.The reforms highlight public welfare nature of public hospitals,which is expected to create profound impacts on hospital operation and medical staff behavior.