Background: Disorders of the blood-brain barrier (BBB) arising from diabetes mellitus are closely related to diabetic encephalopathy. Previous research has suggested that neuron-glia antigen 2 (NG2)-glia plays a key role in maintaining the integrity of the BBB. However, the mechanism by which NG2-glia regulates the diabetic BBB remains unclear. Methods: Type 2 diabetes mellitus (T2DM) db/db mice and db/m mice were used. Evans-Blue BBB permeability tests and transmission electron microscopy techniques were applied. Tight junction proteins were assessed by immunofluorescence and transmission electron microscopy. NG2-glia number and signaling pathways were evaluated by immunofluorescence. Detection of matrix metalloproteinase-9 (MMP-9) in serum was performed using enzyme-linked immunosorbent assay (ELISA). Results: In T2DM db/db mice, BBB permeability in the hippocampus significantly increased from 16 weeks of age, and the structure of tight junction proteins changed. The number of NG2-glia in the hippocampus of db/db mice increased around microvessels from 12 weeks of age. Concurrently, the expression of MMP-9 increased in the hippocampus with no change in serum. Sixteen- week-old db/db mice showed activation of the Wnt/β-catenin signaling in hippocampal NG2-glia. Treatment with XAV-939 improved structural and functional changes in the hippocampal BBB and reduced MMP-9 secretion by hippocampal NG2-glia in db/db mice. It was also found that the upregulation of β-catenin protein in NG2-glia in the hippocampus of 16-week-old db/db mice was significantly alleviated by treatment with XAV-939. Conclusion The results indicate that NG2-glia can lead to structural and functional disruption of the diabetic BBB by activating Wnt/β-catenin signaling, upregulating MMP-9, and degrading tight junction proteins.

Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*

Objective To investigate the population density and seasonal fluctuations of Aedes albopictus in Guangzhou City, Guangdong Province, from 2021 to 2023, so as to provide insights into A. albopictus control and management of dengue fever. Methods The surveillance of A. albopictus density was performed in all surveillance sites assigned across all streets (townships) in Guangzhou City during the period from January to December from 2021 to 2023. The surveillance frequency was twice every half month from May to September, and once every month for the rest of a year. In each surveillance period, A. albopictus mosquito larvae were captured from indoor and outdoor small water containers in residential areas, parks, medical facilities, schools, other government sectors and social organizations, construction sites, special industries and others for mosquito species identification. Adult mosquitoes were captured using electric mosquito suction apparatus for species identification and gender classification. Adult mosquitoes and mosquito eggs were collected with mosquito and egg traps at the breeding and dwelling places of Aedes mosquitoes for identification. The mosquito oviposition index (MOI), Breteau index (BI), adult mosquito density index (ADI) and standard space index (SSI) were calculated. The A. albopictus density was classified into grades 0, 1, 2 and 3 in each surveillance site, with Grade 0 density defined eligible, and the eligible rate of A. albopictus density was calculated at all surveillance sites each year from 2021 to 2023. In addition, the changing trends in MOI, SSI, BI and ADI of A. albopictus were analyzed in Guangzhou City from 2021 to 2023. Results The eligible rates of A. albopictus density were 61.69%, 68.75% and 55.15% in surveillance sites of Guangzhou City from 2021 to 2023 (χ² = 297.712, P < 0.001), and appeared a tendency towards a reduction followed by a rise each year, which gradually reduced since January, maintained at a low level during the period between May and October, and gradually increased from November to December. The MOI, SSI, BI and ADI of A. albopictus all appeared a tendency towards a rise followed by a reduction in Guangzhou City during the period between January and December from 2021 to 2023. The BI of A. albopictus peaked in the first half of June in 2021 (4.03), the first half of July in 2022 (3.89) and the last half of August in 2023 (5.02), and the SSI of A. albopictus peaked in the last half of June in 2021 (0.93), the last half of May in 2022 (0.59), and the last half of June (0.94) and the first half of September in 2023 (1.12). In addition, the MOI of A. albopictus peaked in the first half of May in 2021 (8.64), the first half of June in 2022 (8.96), and the last half of May (10.21) and the last half of June in 2023 (10.89), and the ADI of A. albopictus peaked in the first half of June in 2021 (3.41), the last half of June in 2022 (4.06), and the first half of July in 2023 (3.61). Conclusions The density of A. albopictus is high in Guangzhou City during the period from May to October, and the risk of local outbreak caused by imported dengue fever is high. Persistent intensified surveillance of the density and seasonal fluctuation of A. albopictus is recommended and timely mosquito prevention and control is required according to the fluctuation in the A. albopictus density.

Objective:To investigate the key technical aspects of laparoscopic anti-reflux surgery (LARS) in complex gastroesophageal reflux disease (GERD) with hiatal hernia (HH) and evaluate the feasibility and efficacy of reoperation.Methods:A retrospective analysis was conducted on 28 patients with complex GERD treated at Zhejiang Provincial People's Hospital from Feb 2020 to May 2024. Preoperative examinations were recorded, and surgical videos were reviewed to reconstruct operative time, critical intraoperative steps, complications, and management techniques. Postoperative follow-up via telephone and outpatient visits assessed symptom relief, complications, and medication use.Results:All 28 patients (4 robotic-assisted and 24 conventional laparoscopic surgeries) successfully underwent LARS, with an operative duration of (152.6±10.3) minutes and a postoperative hospital stay of (4.0±1.9) days. Large HH 9 cases, intraoperative bleeding 6 cases, pleural rupture 3 case, and esophageal perforation 1 case, preoperative diagnoses included short esophagus 2 cases and 7 redo surgeries. The overall recurrence rate was 11%. Postoperative complications occurred in 14%. The redo surgeries group achieved 71% symptom resolution. At 1-12 months of follow-up, 82% of patients were asymptomatic, and 82% discontinued proton pump inhibitor therapy.Conclusions:Complex scenarios requiring specialized techniques in LARS increase surgical difficulty and risks. Standardized management of the hernia sac, hiatal repair, neurovascular protection, identification of anatomical landmarks in reoperations, selection of biological mesh, and adhesiolysis may reduce recurrence rates and complication risks.

Acute ischemic stroke is less likely to be caused by carotid artery occlusion related to compression from the hyoid bone.This case report described a patient with acute ischemic stroke due to hyoid bone compression-induced carotid artery occlusion who presented with speech disorder and hemiplegia,without atherosclerotic factors in the past and with exercise history.Head CT angiography showed no atherosclerosis changes in regions outside the offending vessel.Considering the anatomical relationship between hyoid bone and carotid artery,the most reasonable mechanism might be owed to repetitive mechanical compression from ipsilateral greater horn of hyoid bone.It induced endothelial damage to the carotid artery,leading to occlusion and ischemic stroke consequently.It is extremely rare in patients with ischemic stroke.Accordingly,based on the literature review,this study was conducted to explore clinical and imaging manifestations,pathogenesis,diagnosis,and treatment of this special clinical manifestation.

Objective:To investigate the key technical aspects of laparoscopic anti-reflux surgery (LARS) in complex gastroesophageal reflux disease (GERD) with hiatal hernia (HH) and evaluate the feasibility and efficacy of reoperation.Methods:A retrospective analysis was conducted on 28 patients with complex GERD treated at Zhejiang Provincial People's Hospital from Feb 2020 to May 2024. Preoperative examinations were recorded, and surgical videos were reviewed to reconstruct operative time, critical intraoperative steps, complications, and management techniques. Postoperative follow-up via telephone and outpatient visits assessed symptom relief, complications, and medication use.Results:All 28 patients (4 robotic-assisted and 24 conventional laparoscopic surgeries) successfully underwent LARS, with an operative duration of (152.6±10.3) minutes and a postoperative hospital stay of (4.0±1.9) days. Large HH 9 cases, intraoperative bleeding 6 cases, pleural rupture 3 case, and esophageal perforation 1 case, preoperative diagnoses included short esophagus 2 cases and 7 redo surgeries. The overall recurrence rate was 11%. Postoperative complications occurred in 14%. The redo surgeries group achieved 71% symptom resolution. At 1-12 months of follow-up, 82% of patients were asymptomatic, and 82% discontinued proton pump inhibitor therapy.Conclusions:Complex scenarios requiring specialized techniques in LARS increase surgical difficulty and risks. Standardized management of the hernia sac, hiatal repair, neurovascular protection, identification of anatomical landmarks in reoperations, selection of biological mesh, and adhesiolysis may reduce recurrence rates and complication risks.

Acute ischemic stroke is less likely to be caused by carotid artery occlusion related to compression from the hyoid bone.This case report described a patient with acute ischemic stroke due to hyoid bone compression-induced carotid artery occlusion who presented with speech disorder and hemiplegia,without atherosclerotic factors in the past and with exercise history.Head CT angiography showed no atherosclerosis changes in regions outside the offending vessel.Considering the anatomical relationship between hyoid bone and carotid artery,the most reasonable mechanism might be owed to repetitive mechanical compression from ipsilateral greater horn of hyoid bone.It induced endothelial damage to the carotid artery,leading to occlusion and ischemic stroke consequently.It is extremely rare in patients with ischemic stroke.Accordingly,based on the literature review,this study was conducted to explore clinical and imaging manifestations,pathogenesis,diagnosis,and treatment of this special clinical manifestation.

Objective:To analyze the epidemiological and etiological characteristics of a death case of meningococcal meningitis in Hengyang city, Hunan Province in 2024.Methods:Epidemiological investigation of the death case was performed, and samples from the patient and close contacts were collected. Following cultivation and isolation, Neisseria meningitidis ( Nm) strains were analyzed by antimicrobial susceptibility testing, pulsed field gel electrophoresis (PFGE), and whole-genome sequencing for analyzing epidemiological and etiological characteristics. Phylogenetic analysis was carried out using core genomic multilocus sequence typing (cgMLST). Results:The case was a 16-year-old high school boarding student with fulminant meningococcal meningitis. He had shock symptoms, and died within 24 h of the onset of symptoms. Six Nm strains were isolated from the patient and his roommates, belonging to two distinct clades. Isolate 144569 from the patient was highly homologous to isolate 144572 from a close contact, both belonging to the highly pathogenic sublineage L44.1 of CC4821. The typical molecular features was C: P1.7-2, 14: F5-101: ST4821 (CC4821). The two strains carried the antimicrobial resistance genes of gyrA-71 and penA-552, indicating reduced susceptibility to quinolone and penicillin, which was with their resistance phenotype. The isolates from four close contacts clustered within the same clade, characterized by the molecular features of B: P1.18-25, 9-18: ST5829 (UA). Conclusions:The death case is caused by Nm serogroup C from highly pathogenic sublineage L44.1 of CC4821. The spread of this isolate has the potential risk of outbreaks of invasive meningococcal disease. It is necessary to enhanced the molecular epidemiological surveillance, particularly focusing on the transmission of multiple serogroups of Nm among adolescents and the increasing exposure risk.

Periodontitis is a critical risk factor for the occurrence and development of diabetes.Porphyromonas gingivalis may participate in insulin resistance(IR)caused by periodontal inflammation,but the functional role and specific mechanisms of P.gingivalis in IR remain unclear.In the present study,clinical samples were analysed to determine the statistical correlation between P.gingivalis and IR occurrence.Through culturing of hepatocytes,myocytes,and adipocytes,and feeding mice P.gingivalis orally,the functional correlation between P.gingivalis and IR occurrence was further studied both in vitro and in vivo.Clinical data suggested that the amount of P.gingivalis isolated was correlated with the Homeostatic Model Assessment for IR score.In vitro studies suggested that coculture with P.gingivalis decreased glucose uptake and insulin receptor(INSR)protein expression in hepatocytes,myocytes,and adipocytes.Mice fed P.gingivalis tended to undergo IR.P.gingivalis was detectable in the liver,skeletal muscle,and adipose tissue of experimental mice.The distribution sites of gingipain coincided with the downregulation of INSR.Gingipain proteolysed the functional insulin-binding region of INSR.Coculture with P.gingivalis significantly decreased the INSR-insulin binding ability.Knocking out gingipain from P.gingivalis alleviated the negative effects of P.gingivalis on IR in vivo.Taken together,these findings indicate that distantly migrated P.gingivalis may directly proteolytically degrade INSR through gingipain,thereby leading to IR.The results provide a new strategy for preventing diabetes by targeting periodontal pathogens and provide new ideas for exploring novel mechanisms by which periodontal inflammation affects the systemic metabolic state.