Periodontal bone defects, primarily caused by periodontitis, are highly prevalent in clinical settings and manifest as bone fenestration, dehiscence, or attachment loss, presenting a significant challenge to oral health. In regenerative medicine, harnessing developmental principles for tissue repair offers promising therapeutic potential. Of particular interest is the condensation of progenitor cells, an essential event in organogenesis that has inspired clinically effective cell aggregation approaches in dental regeneration. However, the precise cellular coordination mechanisms during condensation and regeneration remain elusive. Here, taking the tooth as a model organ, we employed single-cell RNA sequencing to dissect the cellular composition and heterogeneity of human dental follicle and dental papilla, revealing a distinct Platelet-derived growth factor receptor alpha (PDGFRA) mesenchymal stem/stromal cell (MSC) population with remarkable odontogenic potential. Interestingly, a reciprocal paracrine interaction between PDGFRA⁺ dental follicle stem cells (DFSCs) and CD31⁺ Endomucin⁺ endothelial cells (ECs) was mediated by Vascular endothelial growth factor A (VEGFA) and Platelet-derived growth factor subunit BB (PDGFBB). This crosstalk not only maintains the functionality of PDGFRA⁺ DFSCs but also drives specialized angiogenesis. In vivo periodontal bone regeneration experiments further reveal that communication between PDGFRA⁺ DFSC aggregates and recipient ECs is essential for effective angiogenic-osteogenic coupling and rapid tissue repair. Collectively, our results unravel the importance of MSC-EC crosstalk mediated by the VEGFA and PDGFBB-PDGFRA reciprocal signaling in orchestrating angiogenesis and osteogenesis. These findings not only establish a framework for deciphering and promoting periodontal bone regeneration in potential clinical applications but also offer insights for future therapeutic strategies in dental or broader regenerative medicine.
Receptor, Platelet-Derived Growth Factor alpha/metabolism* ; Humans ; Neovascularization, Physiologic/physiology* ; Dental Sac/cytology* ; Single-Cell Analysis ; Transcriptome ; Mesenchymal Stem Cells/metabolism* ; Bone Regeneration ; Animals ; Dental Papilla/cytology* ; Periodontium/physiology* ; Stem Cells/metabolism* ; Regeneration ; Angiogenesis

Objective:To evaluate physicochemical and immune efficacy of filamentous hemagglutinin(FHA)and pertactin(PRN)before and after detoxification treatment in acellular pertussis vaccine.Methods:Thermal stability of FHA and PRN was analyzed by differential scanning fluorescence(DSF)to evaluate melting temperature(Tm)changes of antigen before and after detoxi-fication treatment.Immune efficacy of vaccine containing single FHA or PRN component and acellular pertussis vaccine with pertussis toxin(PT)was assessed by modified intracerebral challenge assay(MICA)and pertussis serological potency test(PSPT).ELISpot and flow cytometry were utilized to evaluate cytokines secretion and immune cells level in spleens obtained from immunized mice.Results:Under detoxification treatment Tm of FHA changed from a single value 57℃to both 58℃and 81℃,but Tm of PRN were both 84℃.Detoxification treatment had no obvious effects on immune efficacy and IL-4/IFN-γ secretion in spleens from mice.In addi-tion,there was no significant difference in splenic immune cells such as CD4+T,CD8+T and CD44+CD62L+memory cells.Conclu-sion:Detoxification treatment of FHA and PRN have no effect on immune efficacy of acellular pertussis vaccine,except for impact on Tm of FHA.

Objective To investigate the clinical and MRI imaging features of ovarian endometrioid adenofibroma and to analyze its pathological features in order to promote the diagnostic and differential diagnostic ability.Methods The clinical and MRI imaging features of 5 cases of ovarian endometrioid adenofibroma confirmed by pathology were analyzed retrospectively.Results Among the 5 patients,2 were diagnosed with postmenopausal vaginal bleeding,while the other 3 cases were incidental findings with no obvious clinical discomfort.One patient had elevated estrogen level,but the remaining patients showed no obvious abnormalities in laboratory examinations.All the lesions in 5 patients were cystic-solid masses,MRI showed mixed iso-and hypointensity on T1 WI,and slightly hyperintensity and hypointensity of solid components on T2WI.The solid components showed scattered cystic foci with smooth cyst walls and partial septa.The signal in the capsule was homogeneous.Diffusion weighted imaging(DWI)showed slightly hyperintensity and hypointensity.Contrast-enhanced scans showed gradual enhancement in the solid part of the massed with no enhancement in the cystic areas.Conclusion The clinical manifestations of ovarian endometrioid adenofibroma are non-specific,but MRI manifestations are specific to some extent.The final diagnosis depends on pathological diagnosis.

Objective:To evaluate physicochemical and immune efficacy of filamentous hemagglutinin(FHA)and pertactin(PRN)before and after detoxification treatment in acellular pertussis vaccine.Methods:Thermal stability of FHA and PRN was analyzed by differential scanning fluorescence(DSF)to evaluate melting temperature(Tm)changes of antigen before and after detoxi-fication treatment.Immune efficacy of vaccine containing single FHA or PRN component and acellular pertussis vaccine with pertussis toxin(PT)was assessed by modified intracerebral challenge assay(MICA)and pertussis serological potency test(PSPT).ELISpot and flow cytometry were utilized to evaluate cytokines secretion and immune cells level in spleens obtained from immunized mice.Results:Under detoxification treatment Tm of FHA changed from a single value 57℃to both 58℃and 81℃,but Tm of PRN were both 84℃.Detoxification treatment had no obvious effects on immune efficacy and IL-4/IFN-γ secretion in spleens from mice.In addi-tion,there was no significant difference in splenic immune cells such as CD4+T,CD8+T and CD44+CD62L+memory cells.Conclu-sion:Detoxification treatment of FHA and PRN have no effect on immune efficacy of acellular pertussis vaccine,except for impact on Tm of FHA.

Objective To investigate the clinical and MRI imaging features of ovarian endometrioid adenofibroma and to analyze its pathological features in order to promote the diagnostic and differential diagnostic ability.Methods The clinical and MRI imaging features of 5 cases of ovarian endometrioid adenofibroma confirmed by pathology were analyzed retrospectively.Results Among the 5 patients,2 were diagnosed with postmenopausal vaginal bleeding,while the other 3 cases were incidental findings with no obvious clinical discomfort.One patient had elevated estrogen level,but the remaining patients showed no obvious abnormalities in laboratory examinations.All the lesions in 5 patients were cystic-solid masses,MRI showed mixed iso-and hypointensity on T1 WI,and slightly hyperintensity and hypointensity of solid components on T2WI.The solid components showed scattered cystic foci with smooth cyst walls and partial septa.The signal in the capsule was homogeneous.Diffusion weighted imaging(DWI)showed slightly hyperintensity and hypointensity.Contrast-enhanced scans showed gradual enhancement in the solid part of the massed with no enhancement in the cystic areas.Conclusion The clinical manifestations of ovarian endometrioid adenofibroma are non-specific,but MRI manifestations are specific to some extent.The final diagnosis depends on pathological diagnosis.

Objective To investigate the effect of asiaticoside on blood pressure and relaxation of thoracic aorta in rats and explore the underlying mechanism.Methods SD rats treated with 50 and 100 mg/kg asiaticoside by daily gavage for 2 weeks were monitored for systolic blood pressure changes,and histological changes of the thoracic aorta were evaluated using HE staining.In isolated rat endothelium-intact and endothelium-denuded thoracic aorta rings,the effects of asiaticoside on relaxation of the aortic rings were tested at baseline and following norepinephrine(NE)-and KCl-induced constriction.The vascular relaxation effect of asiaticoside was further observed in NE-stimulated endothelium-intact rat aortic rings pretreated with L-nitroarginine methyl ester,indomethacin,zinc protoporphyrin Ⅸ,tetraethyl ammonium chloride,glibenclamide,barium chloride,Iberiotoxin,4-aminopyridine,or TASK-1-IN-1.The aortic rings were treated with KCl and NE followed by increasing concentrations of CaCl2 to investigate the effect of asiaticoside on vasoconstriction induced by external calcium influx and internal calcium release.Results Asiaticoside at 50 and 100 mg/kg significantly lowered systolic blood pressure in rats without affecting the thoracic aorta histomorphology.While not obviously affecting resting aortic rings with intact endothelium,asiaticoside at 100 mg/kg induced significant relaxation of the rings constricted by KCl and NE,but its effects differed between endothelium-intact and endothelium-denuded rings.In endothelium-intact aortic rings pretreated with indomethacin,ZnPP Ⅸ,barium chloride,glyburide,TASK-1-IN-1 and 4-aminopyridine,asiaticoside did not produce significant effect on NE-induced vasoconstriction,and tetraethylammonium,Iberiotoxin and L-nitroarginine methyl ester all inhibited the relaxation effect of asiaticoside.In KCl-and NE-treated rings,asiaticoside obviously inhibited CaCl2-induced vascular contraction.Conclusion Asiaticoside induces thoracic aorta relaxation by mediating high-conductance calcium-activated potassium channel opening,promoting nitric oxide release from endothelial cells and regulating Ca2+ influx and outflow,thereby reducing systolic blood pressure in rats.

OBJECTIVE To analyze the cost-effectiveness of abemaciclib， palbociclib and ribociclib combined with aromatase inhibitors （AI） in first-line treatment of hormone receptor-positive （HR+） advanced breast cancer from the perspective of Chinese medical system. METHODS The 20-year disease course of the patients was simulated by the partitioned survival model， and the simulation period was determined to be 4 weeks， the model output was the total cost and quality-adjusted life year （QALY）， the cost and effect were discounted at a discount rate of 5%. A network meta-analysis was constructed by systematically searching relevant clinical trials to obtain the efficacy parameters of abemaciclib， palbociclib and ribociclib combined with AI. Survival fitting and extrapolation were performed based on the survival curve of the placebo group in MONALEESA-2 trial. Cost-effectiveness was assessed by incremental cost-effectiveness ratio （ICER） and incremental net monetary benefit （INMB）， with a willingness-to-pay threshold of 3 times China’s per capita gross domestic product （GDP） in 2023； one-way sensitivity analysis and probability sensitivity analysis were used to detect the influence of parameters on the results and the robustness of the incremental analysis results. RESULTS In the 20-year simulation， compared with palbociclib+AI scheme， the ICER of ribociclib+AI scheme was 58 558.38 yuan/QALY and the INMB was 62 988.20 yuan. Compared with ribociclib+AI scheme， the ICER of abemaciclib+AI scheme was 264 928.34 yuan/QALY and the INMB was 344.84 yuan. One-way sensitivity analysis showed that the incremental analysis results of abemaciclib+AI scheme compared to ribociclib+AI scheme were not robust. Probabilistic sensitivity analysis confirmed that the probability of ribociclib+AI scheme becoming the most economical was the highest when the threshold was 1-3 times China’s per capita GDP in 2023. CONCLUSIONS Ribociclib+AI scheme is more likely to be the most economical first-line treatment than abemaciclib+AI scheme and palbociclib+AI scheme in Chinese patients with HR+ advanced breast cancer when threshold is 1-3 times China’s per capita GDP in 2023.

Objective To investigate the effect of asiaticoside on blood pressure and relaxation of thoracic aorta in rats and explore the underlying mechanism.Methods SD rats treated with 50 and 100 mg/kg asiaticoside by daily gavage for 2 weeks were monitored for systolic blood pressure changes,and histological changes of the thoracic aorta were evaluated using HE staining.In isolated rat endothelium-intact and endothelium-denuded thoracic aorta rings,the effects of asiaticoside on relaxation of the aortic rings were tested at baseline and following norepinephrine(NE)-and KCl-induced constriction.The vascular relaxation effect of asiaticoside was further observed in NE-stimulated endothelium-intact rat aortic rings pretreated with L-nitroarginine methyl ester,indomethacin,zinc protoporphyrin Ⅸ,tetraethyl ammonium chloride,glibenclamide,barium chloride,Iberiotoxin,4-aminopyridine,or TASK-1-IN-1.The aortic rings were treated with KCl and NE followed by increasing concentrations of CaCl2 to investigate the effect of asiaticoside on vasoconstriction induced by external calcium influx and internal calcium release.Results Asiaticoside at 50 and 100 mg/kg significantly lowered systolic blood pressure in rats without affecting the thoracic aorta histomorphology.While not obviously affecting resting aortic rings with intact endothelium,asiaticoside at 100 mg/kg induced significant relaxation of the rings constricted by KCl and NE,but its effects differed between endothelium-intact and endothelium-denuded rings.In endothelium-intact aortic rings pretreated with indomethacin,ZnPP Ⅸ,barium chloride,glyburide,TASK-1-IN-1 and 4-aminopyridine,asiaticoside did not produce significant effect on NE-induced vasoconstriction,and tetraethylammonium,Iberiotoxin and L-nitroarginine methyl ester all inhibited the relaxation effect of asiaticoside.In KCl-and NE-treated rings,asiaticoside obviously inhibited CaCl2-induced vascular contraction.Conclusion Asiaticoside induces thoracic aorta relaxation by mediating high-conductance calcium-activated potassium channel opening,promoting nitric oxide release from endothelial cells and regulating Ca2+ influx and outflow,thereby reducing systolic blood pressure in rats.

Objective:To investigate the expression of CD133(also known as PROM1)in pancreatic cancer tissues and its association with clinicopathological features and prognosis of pancreatic cancer patients.Methods:The GEPIA website was used to analyze the expression of CD133 in pancreatic cancer patients from the TCGA database.The correlation between PROM1 mRNA expression and cancer stem cell family genes in human pancreatic cancer tissues was analyzed based on the TCGA database.The expression of CD133 in pancreatic cancer tissues and its clinical significance were studied by immunohistochemistry(IHC)staining.Wilcoxon rank sum test was used to analyze the difference in CD133 expression between pancreatic cancer tissues and adjacent tissues.Chi-square test was used to analyze the relationship between CD133 expression and clinicopathological characteristics in pancreatic cancer tissues.Kaplan-Meier method and Log-rank test were used to analyze the survival difference based on different levels of CD133 expression in pancreatic cancer tissues.The Cox model was used to evaluate the prognostic value of different indicators.Hazard ratio(HR)and 95%confidence interval(95%CI)were used to assess the strength of the association between CD133 expression and mortality risk in pancreatic cancer patients.Results:TCGA database analysis showed that the expression of CD133 was significantly up-regulated in pancreatic cancer tissues compared with adjacent tissues(P<0.05).The mRNA expression level of PROM1 in pancreatic cancer tissues was correlated with tumor stem cell family genes,including EPCAM,POU5F1,CD24,CD44 and CXCR4.The expression level of CD133 in pancreatic cancer tissues was significantly associated with tumor differentiation,TNM stage,and lymph node metastasis(all P<0.05).Univariate Cox model analysis showed that overall survival(OS)was significantly associated with age(HR=0.544,95%CI[0.299,0.990],P<0.05),depth of invasion(HR=0.496,95%CI[0.292,0.842],P<0.05),TNM stage(HR=2.148,95%CI[1.352,3.412],P<0.05),and CD133 expression(HR=1.935,95%CI[1.090,3.433],P<0.05).Multivariate Cox model analysis showed that TNM stage(HR=0.116,95%CI[0.025,0.551],P<0.05),lymph node metastasis(HR=0.392,95%CI[0.160,0.960],P<0.05)and CD133 expression(HR=2.080,95%CI[1.053,4.106],P<0.05)were independent prognostic risk factors.Conclusion:CD133 is highly expressed in pancreatic cancer tissues,and its expression is significantly associated with the prognosis of pancreatic cancer patients.CD133 may serve as a potential new target for immunotherapy in pancreatic cancer.

ObjectiveTo investigate the intervention effect of Buzhong Yiqitang （BZYQT） on pulmonary inflammation in mice induced by chronic intermittent hypoxia （CIH） and preliminarily elucidate its mechanism. MethodForty healthy male C57BL/6 mice aged 6-8 weeks were randomly divided into the following groups： normoxia group， model group （exposed to CIH）， and low-， medium-， and high-dose BZYQT groups. The normoxia group was exposed to a normoxic environment， while the model group and the low-， medium-， and high-dose BZYQT groups were exposed to intermittent hypoxia. In the BZYQT groups， the BZYQT （8.1， 16.2， 32.4 g·kg^-1·d^-1） was administered orally 30 min before placing the mice in the hypoxic chamber， while the model group and the normoxia group received an equivalent volume of normal saline. After five weeks of modeling， pulmonary function of the mice was measured using an EMKA animal lung function analyzer， and lung tissue samples were collected after the pulmonary function tests. Hematoxylin-eosin （HE） staining was performed to observe the histopathological changes in the lung tissue of each group. Enzyme-linked immunosorbent assay （ELISA） was used to measure the levels of interleukin-6 （IL-6）， interleukin-8 （IL-8）， tumor necrosis factor-α （TNF-α） in the serum， as well as angiotensin Ⅱ （Ang Ⅱ） and angiotensin-（1-7） ［Ang（1-7）］ in lung tissue. Western blot and immunohistochemistry were used to detect the protein expression of IL-6， IL-8， TNF-α， angiotensin-converting enzyme 2 （ACE2）， and mitochondrial assembly receptor （Mas）. ResultCompared with the normoxia group， the model group showed significant abnormalities in lung function （P<0.05， P<0.01）， lung tissue changes， such as thickening of alveolar walls and inflammatory cell infiltration， increased levels of IL-6， IL-8， TNF-α in the serum and Ang Ⅱ in lung tissue （P<0.01）， decreased level of Ang（1-7） （P<0.01）， increased protein expression of IL-6， IL-8， and TNF-α， and decreased protein expression of ACE2 and Mas （P<0.05， P<0.01）. Compared with the model group， the BZYQT groups showed improvement in lung function （P<0.05， P<0.01）， and HE staining of lung tissue showed approximately normal alveolar wall thickness and reduced inflammatory cell infiltration. Immunohistochemistry and Western blot analysis showed a significant decrease in the expression of inflammatory-related proteins （P<0.05， P<0.01）， and a significant increase in ACE2 and Mas protein expression （P<0.05， P<0.01）. ConclusionBZYQT can improve lung injury in mice exposed to CIH by regulating the ACE2-Ang（1-7）-Mas axis to inhibit inflammatory responses.