Complement C3 plays a critical role in periodontitis. However, its source, role and underlying mechanisms remain unclear. In our study, by analyzing single-cell sequencing data from mouse model of periodontitis, we identified that C3 is primarily derived from periodontal fibroblasts. Subsequently, we demonstrated that C3a has a detrimental effect in ligature-induced periodontitis. C3ar^-/- mice exhibited significantly less destruction of periodontal support tissues compared to wild-type mice, characterized by mild gingival tissue damage and reduced alveolar bone loss. This reduction was associated with decreased production of pro-inflammatory mediators and reduced osteoclast infiltration in the periodontal tissues. Mechanistic studies suggested that C3a could promote macrophage polarization and osteoclast differentiation. Finally, by analyzing single-cell sequencing data from the periodontal tissues of patients with periodontitis, we found that the results observed in mice were consistent with human data. Therefore, our findings clearly demonstrate the destructive role of fibroblast-derived C3 in ligature-induced periodontitis, driven by macrophage M1 polarization and osteoclast differentiation. These data strongly support the feasibility of C3a-targeted interventions for the treatment of human periodontitis.
Animals ; Osteoclasts/cytology* ; Periodontitis/metabolism* ; Cell Differentiation ; Mice ; Fibroblasts/metabolism* ; Macrophages ; Disease Models, Animal ; Complement C3/metabolism* ; Humans ; Disease Progression ; Mice, Inbred C57BL ; Male ; Mice, Knockout

Objective·To observe the effects of gingipain extract on the biological characteristics of oral squamous cell carcinoma cell HN6.Methods·The HN6 cell line was selected,cultivated,and divided into different groups based on the protein concentration of gingipain extract from Porphyromonas gingivalis:control group,3.125 μg/mL group,6.25 μg/mL group,12.5 μg/mL group,25 μg/mL group,50 μg/mL group,and 100 μg/mL group.After 24 and 48 h of cultivation,CCK-8 assay was used to detect the effects of gingipain extract on HN6 cell proliferation activity.Subsequent experiments were divided into control group,25 μg/mL group and 50 μg/mL group.Flow cytometry was used to examine the effects of gingipain extract on cell cycle.Scratch assay and Transwell assay were performed to evaluate cell migration and invasion ability.Real-time PCR(RT-PCR)and Western blotting were used to measure the expression of E-cadherin and N-cadherin proteins and genes in cells.Results·Stimulated with gingipain extract for 24 h,the HN6 cells showed significantly increased proliferation activity in the 25 μg/mL(P=0.025),50 μg/mL(P=0.000),and 100 μg/mL(P=0.049)groups compared to the control group.After 48 h,proliferation activity was significantly higher in the 6.25 μg/mL(P=0.024),12.5 μg/mL(P=0.006),25 μg/mL(P=0.000),50 μg/mL(P=0.000),and 100 μg/mL(P=0.000)groups compared to the control group.Cell cycle analysis revealed that,after 24 h of gingipain stimulation,the proportion of HN6 cells in the G1 phase decreased,while the proportion in the S+G2 phase significantly increased compared to the control group(25 μg/mL group:P=0.024;50 μg/mL group:P=0.001).Compared to the control group,the scratch assay demonstrated a significant increase in the percentage of scratch closure as the concentration of gingipain extract increased(P=0.001).Compared to the control group,the Transwell invasion assay showed a significant increase in the number of cells passing through the bottom of the chamber as the concentration of gingipain extract increased.RT-PCR and Western blotting results indicated that as the concentration of gingipain extract increased,the expression levels of N-cadherin mRNA and protein in HN6 cells significantly increased,while the expression levels of E-cadherin mRNA and protein significantly decreased compared to the control group.Conclusion·Gingipain extract could promote proliferation,migration,and invasion of oral squamous cell carcinoma HN6 cells.

Pulmonary hypertension has a high mortality rate,and although targeted therapy is available,it is still incurable,and the long-term prognosis for patients is poor.As a tyrosine kinase inhibitor,imatinib was approved for marketing in China in 2002 for the treatment of chronic myelogenous leukemia and other tumor diseases.In addition to the antitumor effects,imatinib was found to improve hemodynamics and exercise tolerance in patients with severe pulmonary arterial hypertension,but the safety was suboptimal.With the emergence of new formulations of imatinib targeted at the lungs,it is expected to become a new targeted drug for pulmonary arterial hypertension.

Purpose To summarize the clinical and imaging features of neonatal herpes simplex encephalitis(NHSE).Materials and Methods Clinical and imaging data of 5 NHSE from January 2016 to June 2023 in Beijing Children's Hospital,Capital Medical University were retrospectively collected.All five children underwent MRI examinations,with three of them undergoing enhanced scanning simultaneously.Two children had previously undergone CT scans.The location,density/signal,enhancement characteristics as well as follow-up imaging changes of the lesions were reviewed.Results The main clinical manifestations of NHSE were fever(5 cases)and seizure(4 cases),sometimes accompanied by herpes(2 cases).Imaging examinations in NHSE typically presented with symmetric(1 case)or diffuse/multifocal(4 cases)lesions in bilateral cerebral hemispheres,along with involvement of the bilateral thalamus(5 cases).Early CT scans showed either no abnormalities(1 case)or extensive areas of low density(1 case).MRI examinations usually demonstrated restricted diffusion of acute phase lesions(3 cases)and significant leptomeningeal enhancement in affected areas(3 cases).Intracranial lesions often led to the diffuse atrophy of brain parenchyma and polycystic encephalomalacia(3 cases),indicating a poor prognosis.Conclusion The clinical manifestations of NHSE are nonspecific.Early MRI examinations are of great value for accurate diagnosis and disease evaluation.

Objective:To analyze high-risk factors for poor neurological prognosis in full-term neonatal purulent meningitis based on clinical and brain MRI features.Methods:This study was a case-control study. The clinical and brain MRI data of 79 neonates with purulent meningitis were retrospectively collected at Beijing Children′s Hospital, Capital Medical University from January 2016 to January 2022. Follow-up assessments including growth and development, as well as neurological sequelae, were conducted over a minimum follow-up period of 6 months. The patients were divided into two groups with good ( n=49) and poor prognosis ( n=30) according to follow-up results. Chi-square tests were used to compare clinical and brain MRI features between the two groups, and a multivariate logistic regression analysis was performed to explore the high-risk factors for poor neurologic prognosis in full-term neonates with purulent meningitis. Results:There were statistically differences between two groups regarding the incidence of seizures, early-onset manifestations, positive cerebrospinal fluid (CSF) culture, CSF white cell counts, and CSF protein concentration ( P<0.05). Statistically differences were also found in the occurrence rates of ependymitis, obvious ventricular dilatation/hydrocephalus, spotty and patchy brain injury/hemorrhage, and destructive lesions within the brain parenchyma ( P<0.05). The results of multivariate logistic regression analysis indicated that seizures ( OR=5.722, 95% CI 1.126-29.072, P=0.035), early-onset neonatal purulent meningitis ( OR=3.657, 95% CI 1.073-12.459, P=0.038), ependymitis ( OR=8.851, 95% CI 1.169-67.017, P=0.035), obvious ventricular dilatation/hydrocephalus ( OR=12.675, 95% CI 1.085-148.110, P=0.043), and destructive lesions within the brain parenchyma ( OR=16.370, 95% CI 1.575-170.175, P=0.019) were independent risk factors for poor prognosis. Conclusions:The occurrence of seizures, early-onset manifestations as well as ependymitis, obvious ventricular dilatation/hydrocephalus, and destructive lesions within the brain parenchyma on MRI are high-risk factors for poor prognosis in the full-term neonate with purulent meningitis.

Objective Genotypes(G)1,3,and 5 of the Japanese encephalitis virus(JEV)have been isolated in China,but the dominant genotype circulating in Chinese coastal areas remains unknown.We searched for G5 JEV-infected cases and attempted to elucidate which JEV genotype was most closely related to human Japanese encephalitis(JE)in the coastal provinces of China. Methods In this study,we collected serum specimens from patients with JE in three coastal provinces of China(Guangdong,Zhejiang,and Shandong)from 2018 to 2020 and conducted JEV cross-neutralization tests against G1,G3,and G5. Results Acute serum specimens from clinically reported JE cases were obtained for laboratory confirmation from hospitals in Shandong(92 patients),Zhejiang(192 patients),and Guangdong(77 patients),China,from 2018 to 2020.Seventy of the 361 serum specimens were laboratory-confirmed to be infected with JEV.Two cases were confirmed to be infected with G1 JEV,32 with G3 JEV,and two with G5 JEV. Conclusion G3 was the primary infection genotype among JE cases with a definite infection genotype,and the infection caused by G5 JEV was confirmed serologically in China.

Objective:To analyze the growth characteristics of different genotypes of Japanese encephalitis virus (JEV) in different cell lines, and to provide scientific basis for the selection of cell lines in the study of JEV.Methods:BHK-21, Vero, C6/36, PK-15, DF-1, N2a, SH-sy5y and MDCK cell lines were selected. The proliferation ability of genotype 1 (NX1889 strain), genotype 3 (P3 strain) and genotype 5 (XZ0934 strain) JEV in these cell lines was evaluated by plaque assay and RT-qPCR.Results:Significant cytopathogenic effects (CPE) were observed in BHK-21, Vero, C6/36, DF-1, N2a and PK-15 cell lines across all three JEV genotypes. However, no significant differences in CPE characteristics were observed within the same cell line. SH-sy5y and MDCK cell lines did not show significant CPE, but virus proliferation was detected in SH-sy5y cell line, while MDCK cell line were found to be insensitive to JEV. No significant difference was observed in the proliferation curves of G1, G3 and G5 JEV in BHK-21, Vero and SH-sy5y cell lines. In C6/36 and PK-15 cell lines, the titer of G1 JEV was higher than that of G3 and G5. In DF-1 cell line, G5 demonstrated a higher titer than the other two genotypes, whereas in N2a cell line, G5 showed a lower titer than the other two.Conclusions:There are differences in the proliferation of three different genotypes of JEV in different cell lines, which can provide reference for the study of JEV in different directions.

Objective To explore the efficacy and safety of different antiplatelet drugs combined with rivaroxaban in patients with lower extremity arteriosclerosis obliterans.Methods The clinical data of patients with lower extremity arteriosclerosis obliterans who were symptomatic and underwent surgical treatment at the Vascular Surgery Department of Nanjing Drum Tower Hospital from January 2018 to December 2021 were retrospectively analyzed.According to the different antiplatelet medications taken by patients,patients were categorized into aspirin group,clopidogrel group and cilostazol group.Baseline data of patients were collected,and patients were followed up and the incidence of major adverse cardiovascular events,major adverse limb events,major bleeding and clinically related non-major bleeding events were compared in different groups.Results A total of 632 patients were included in the study.There was no significant difference in the incidence of major adverse cardiovascular events,major adverse limb events,major bleeding and clinically related non-major bleeding events after the baseline data was balanced by inverse probability of treatment weighting.The results of subgroup analysis were generally consistent with those of the overall study.Conclusion The combination of clopidogrel or cilostazol with rivaroxaban may serve as a novel option for dual antithrombotic therapy in patients diagnosed with lower extremity arteriosclerotic obliterans.

Objective To systematically evaluate brain parenchymal and vascular changes after neonatal arterial ischaemic stroke(AIS)using nonenhanced brain neurovascular MR sequences.Methods MR imaging data of 57 children with AIS(＜28 days old at the time of diagnosis)were analyzed retrospectively,including[susceptibility weighted imaging(SWI),magnetic resonance angiography(MRA),magnetic resonance venography(MRV)]sequences and conventional brain sequences[T1 WI,T2 WI,diffusion weighted imaging(DWI)].The imaging features and alteration trend of different sequences in different periods were summarized and studied.Results Vessels increased or decreased in number and enlarged or reduced in diameter within the infarction area,which presented a mixed-pattern,while the periphery showed increased only by using the nonenhanced brain vascular sequence.During the first 9 days after the onset of symptoms,the signals of T1 WI and T2 WI within the infarction area gradually stabilized,and most signals of DWI returned to normal.MRA,MRV,and SWI sequences showed a minimal change of diameter and number of vessels within the infarction area and its surroundings,which directed that neonatal AIS may had transient and dramatic brain injury and blood flow alteration.Conclusion The nonenhanced neurovascular sequence of brain MR can comprehensively display the alteration of brain parenchymal signals and vascular trends after neonatal AIS,providing the possibility for early clinical rapid and effective evaluation.