ObjectiveMigraine is a leading neurological disorder and the fourth most common cause of years lived with disability worldwide, affecting nearly 116 million individuals. Although pharmacological treatments are available, their efficacy is often limited by side effects and variable response rates. Repetitive transcranial magnetic stimulation (rTMS) over the dorsolateral prefrontal cortex (DLPFC) offers a safe, non-invasive alternative for migraine management. However, the neurophysiological mechanisms, particularly how rTMS modulates local cortical excitability and distributed pain-related circuits, remain poorly understood. Elucidating these mechanisms is essential for optimizing treatment protocols and improving clinical outcomes. MethodsThis study employed concurrent transcranial magnetic stimulation and electroencephalography (TMS-EEG) to investigate neuroplastic and neurocircuitry mechanisms of DLPFC-rTMS in migraine. Study 1 compared 30 migraineurs and 28 healthy controls to identify abnormalities in TMS-evoked potentials (TEPs) and significant current density (SCD) within sensory-discriminative regions including the primary somatosensory cortex (S1) and posterior insula (pINS), cognitive-affective regions including the anterior insula (aINS) and midcingulate cortex (MCC), and a descending modulatory region, the periaqueductal gray (PAG). Study 2 used a single-blind, crossover, sham-controlled design in 34 healthy participants. Each participant received both active (10 Hz, 80% RMT, 1 500 pulses) and sham DLPFC-rTMS in counterbalanced order. TMS-EEG and cold pain tolerance were assessed before and after each session. ResultsIn Study 1, migraineurs showed a significantly less negative N120 amplitude compared to healthy controls (P=0.027, Cohen’s d=0.60), indicating local intracortical disinhibition. No group differences were observed for N40, P60, or P180 components. At the source level, migraineurs exhibited significantly higher SCD in the S1, pINS, aINS, and MCC (allQ<0.05), but not in the ventroposterior thalamus (vpTHAL), mediodorsal thalamus (mdTHAL), or PAG. In Study 2, active rTMS significantly reduced SCD from pre- to post-stimulation in the S1, aINS, and MCC (all Q<0.05). Sham stimulation also reduced SCD in the S1 (Q<0.05) but not in the aINS or MCC. Although no significant group-level analgesic effect was observed between active and sham conditions (P=0.107), correlation analyses revealed that greater SCD reductions in the S1 and MCC were significantly associated with higher post-rTMS pain tolerance (R=-0.487 and -0.495, both Q<0.01) and larger improvements in pain tolerance(R=-0.487 and -0.451, both Q<0.05). No such correlations were found following sham stimulation, suggesting that the behavioural relevance of neural changes is specific to active rTMS. ConclusionThis study provides novel evidence that migraineurs exhibit both local neuroplastic abnormalities (reduced N120 amplitude) and hyperactivity in key pain-processing regions (S1, pINS, aINS, MCC). A single session of DLPFC-rTMS reduced hyperactivity in the aINS, MCC, and S1. Notably, greater reductions in the S1 and MCC were associated with improved pain tolerance. These findings identify distinct cortical circuitries, particularly within the cognitive-affective pain network, that may serve as potential biomarkers for optimizing rTMS treatment in migraine and other chronic pain conditions. Future studies should validate these results in patient populations experiencing spontaneous migraine attacks and explore multi-session or accelerated rTMS protocols.

Objective: To investigate follow-up participation situation, negative conversion, and tuberculosis incidence among boarding junior and senior high school freshmen with latent tuberculosis infection(LTBI) in Zhuhai, so as to provide a reference for better LTBI management in school settings. Methods: A prospective cohort study enrolled 413 LTBI and 27 184 non LTBI boarding junior and senior high school freshmen in Zhuhai from September to October 2023. The LTBI group underwent follow up retesting at >3-6, >6-12, >12-24, >24 months after screening. The followup content included mycobacterium tuberculosis infection testing,chest X ray examination,and questionaire survey(basic demographic information, lifestyle, history of close contact with tuberculosis, and suspected symptoms of pulmonary tuberculosis). Follow-up participation, negative conversion, and tuberculosis incidence were analyzed. Rate was compared by using the Chi-square test, and factors associated with negative conversion were assessed by multivariate Logistic regression analysis. Results: Follow-up participation rates increased initially and then declined, with rates of 25.67% within >3-6 months, 37.05% within >6-12 months, 82.02% within >12-24 months, and 52.21% after 24 months. The overall negative conversion rate was 61.03%. The differences in the conversion rate to negative of mycobacterium tuberculosis infection detection among junior and senior high school freshmen in the LTBI group by different testing methods, school types, smoking history, sleep status, and academic stress were all statistically significant( χ 2=124.96, 22.70, 4.71, 6.31, 6.08, all P <0.05). Multivariate Logistic regression analysis identified testing methods, smoking history, and sleep status as factors associated with negative conversion ( OR =1.90-21.67, all P <0.05). The cumulative incidence of tuberculosis was 726.39 per 100 000 in the LTBI group versus 14.71 per 100 000 in the non-LTBI group ( RR =49.37, 95% CI =11.08-219.87, P <0.01). Conclusions Boarding junior and senior high school freshmen with LTBI in Zhuhai have a higher risk of tuberculosis than concurrent non-LTBI students; they therefore represent a priority group for tuberculosis prevention and control. Centralized follow up is recommended to improve participation, and high specificity methods should be used in follow up retesting and maintain consistency across assessments.

Purpose To investigate the expression of Versican(VCAN)and thrombospondin 2(THBS2)and their relationship with cancer-associated fibroblast(CAFs)and clinicopathological significance in papillary thyroid car-cinoma(PTC).Methods Bioinformatics analyses were performed using PTC single-cell sequencing data from the GEO and TCGA database.Weighted correlation network analysis identified CAFs-related genes,and enrichment analy-sis highlighted pivotal genes associated with CAFs;the expression of VCAN,THBS2,and α-SMA in 130 PTC tissue samples were detected by immunohistochemistry EnVision method.Masson staining evaluated tumor stromal fibrosis.Relationships between these markers,clinicopathological parameters,and CAF proliferation were analyzed.Results Bioinformatics analysis identified VCAN and THBS2 as core genes significantly associated with CAFs,and extracellular matrix-related pathways.The proliferation rate of CAFs in PTC was 83.1％(108/130),with positivity rate of 96.9％(126/130)for VCAN and 75.4％(98/130)for THBS2.The median mesenchymal fibrosis index was 32.4(inter-quartile range:22.7-50.0).High CAF proliferation correlated positively with lymph node metastasis(P＜0.001),higher TNM stage(P＜0.05),and specific histologic subtypes of PTC.Similarly,VCAN expression,THBS2 expres-sion,and the degree of PTC stromal fibrosis were positively correlated with lymph node metastasis(P＜0.001,P＜0.05,and P＜0.001,respectively).Both THBS2 expression and the degree of PTC stromal fibrosis correlated with the histologic subtype of PTC.The percentage of tumor mesenchymal α-SMA-positive cells strongly correlated with the im-mune response score(IRS)of VCAN and THBS2(rs=0.713,P＜0.001;rs=0.646,P＜0.001).Additionally,the percentage of Masson-stained area was positively correlated with the percentage of tumor mesenchymal α-SMA-posi-tive cells,and the IRS of VCAN and THBS2(rs=0.892,P＜0.001;rs=0.729,P＜0.001;rs=0.616,P＜0.001).Conclusion VCAN and THBS2 serve as potential markers for assessing invasiveness and lymph node metas-tasis of PTC.Their strong association with CAFs provides a basis for further investigation of the malignant biological be-havior of PTC.

Objective To investigate the changes of serum microRNA (miR)-138-5p and microRNA(miR)-574-5p expression levels and their clinical significance in newborns acute respiratory distress syndrome (nARDS). Methods A total of 112 infants with nARDS from September 2018 to August 2023 in the Neonatal Department of Xiaogan Hospital Affiliated to Wuhan University of Science and Technology were collected as the observation group. They were divided into good prognosis group (n=98) and the poor prognosis group (n=14). A total of 104 healthy full-term newborns born at the same time were selected as the control group. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-138-5p and miR-574-5p in serum. ROC curve was used to analyze the diagnostic value of serum miR-574-5p and miR-138-5p levels for poor prognosis of infants with nARDS. Multivariate Logistic regression analysis of factors affecting the poor prognosis of infants with nARDS. Results Compared with the control group,the expression level of serum miR-574-5p in the observation group increased (1.72±0.23 vs 1.04±0.17),with the expression level of miR-138-5p decreased (0.55±0.08 vs 1.02±0.16),with statistically significant differences (t=24.557,25.597,all P<0.05). Compared with the good prognosis group,the serum miR-138-5p expression level in the poor prognosis group decreased (0.41±0.06 vs 0.57±0.08),with the miR-574-5p expression level increased (2.07±0.25 vs 1.67±0.19),with statistically significant differences (t=7.188,7.069,all P<0.05). The age,intrauterine distress,and proportion of abnormal amniotic fluid in the poor prognosis group were higher than those in the good prognosis group (t=5.359,4.257,6.577,all P<0.05). MiR-574-5p was an independent risk factor for poor prognosis with nARDS,and miR-138-5p was a protective factor (all P<0.05).The AUC (95％ CI) of serum miR-138-5p and miR-574-5p alone and combined diagnosis of nADRS were 0.793 (0.706～0.864),0.898 (0.826～0.947) and 0.931 (0.867～0.970),respectively. The combined prediction of miR-138-5p and miR-574-5p were better than that of miR-138-5p alone and miR-574-5p alone (Z=2.151,1.982,all P<0.05). Conclusion The expression levels of miR-138-5p in serum of infant with nARDS were lower and the expression levels of miR-574-5p were higher,both of which have certain diagnostic value for poor prognosis of infant with nARDS .

Glycoengineering was carried out in the mammalian cell line CHO for the production of pro-tein-based drugs.Firstly,the genome sequence of the Rosa26 locus of CHO cells was determined,the gRNA sequences were designed,and the landing pad was integrated into the Rosa26 locus of CHO cells by CRISPR/Cas9 technology.Three targeting vectors co-expressed by glycosyltransferases,which are β-1,4 galactosyltransferase(B4GALT1),α-2,6-sialyltransferase 1(ST6GAL1)and N-acetaminoglycosyl-transferase Ⅲ(GnT Ⅲ),were constructed by overlapping PCR and seamless ligation technology,and the three glycosyltransferase genes were integrated into the CHO Rosa26 locus by Cre enzyme-mediated cassette exchange technology.PCR confirmed that three glycosyltransferases had been successfully site-directed integrated into the Rosa26 site.The mRNA expression levels of the three glycosyltransferases were more than 50 000-fold by qRT-PCR,and the protein expression levels of the three glycosyltrans-ferases were more than 4-fold via western blotting(P＜0.001).A CHO-engineered cell line with three glycosyltransferases integrated into Rosa26 site was successfully constructed.

Objective To investigate the changes of serum microRNA (miR)-138-5p and microRNA(miR)-574-5p expression levels and their clinical significance in newborns acute respiratory distress syndrome (nARDS). Methods A total of 112 infants with nARDS from September 2018 to August 2023 in the Neonatal Department of Xiaogan Hospital Affiliated to Wuhan University of Science and Technology were collected as the observation group. They were divided into good prognosis group (n=98) and the poor prognosis group (n=14). A total of 104 healthy full-term newborns born at the same time were selected as the control group. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-138-5p and miR-574-5p in serum. ROC curve was used to analyze the diagnostic value of serum miR-574-5p and miR-138-5p levels for poor prognosis of infants with nARDS. Multivariate Logistic regression analysis of factors affecting the poor prognosis of infants with nARDS. Results Compared with the control group,the expression level of serum miR-574-5p in the observation group increased (1.72±0.23 vs 1.04±0.17),with the expression level of miR-138-5p decreased (0.55±0.08 vs 1.02±0.16),with statistically significant differences (t=24.557,25.597,all P<0.05). Compared with the good prognosis group,the serum miR-138-5p expression level in the poor prognosis group decreased (0.41±0.06 vs 0.57±0.08),with the miR-574-5p expression level increased (2.07±0.25 vs 1.67±0.19),with statistically significant differences (t=7.188,7.069,all P<0.05). The age,intrauterine distress,and proportion of abnormal amniotic fluid in the poor prognosis group were higher than those in the good prognosis group (t=5.359,4.257,6.577,all P<0.05). MiR-574-5p was an independent risk factor for poor prognosis with nARDS,and miR-138-5p was a protective factor (all P<0.05).The AUC (95％ CI) of serum miR-138-5p and miR-574-5p alone and combined diagnosis of nADRS were 0.793 (0.706～0.864),0.898 (0.826～0.947) and 0.931 (0.867～0.970),respectively. The combined prediction of miR-138-5p and miR-574-5p were better than that of miR-138-5p alone and miR-574-5p alone (Z=2.151,1.982,all P<0.05). Conclusion The expression levels of miR-138-5p in serum of infant with nARDS were lower and the expression levels of miR-574-5p were higher,both of which have certain diagnostic value for poor prognosis of infant with nARDS .

Objective To develop an automated system that can accurately determine the relationship between the mandibular third molar and the mandibular nerve canal from panoramic images.Methods A dataset consisting of 600 panoramic images of the oral cavi-ty was selected,and the positions of the mandibular third molar and the mandibular nerve canal were accurately labeled.We compared the research designed TI-YOLOv5 with PANet,Faster R-CNN,Mask R-CNN,ResNeSt-101,and the original YOLOv5 in image seg-mentation tasks,with evaluation metrics of AP and AP50.Results TI-YOLOv5 achieved AP(average precision)54.0％and AP5094.9％,an increase of 4.9 and 6.7 percentage points respectively compared to the original YOLOv5(AP 49.1％,AP50 88.2％),and surpassed other SOTA methods such as Mask R-CNN(AP 45.1％,AP50 84.2％).Conclusion TI-YOLOv5 is significantly superior to mainstream networks in automatic positioning and relationship classification of mandibular wisdom teeth and neural tubes,with high de-tection accuracy and discrimination accuracy,and can provide reliable technical support for preoperative risk assessment of mandibular wisdom tooth extraction.

AIM To investigate the stability of salvianolic acid B.METHODS HPLC was adopted in the content determination of salvianolic acid B,after which the chemical stability in different pH of buffer solutions,oxidation stability in different concentrations of H2O2,and biological stability in artificial gastric fluid,artificial intestinal fluid and biological matrices were analyzed,and its degradation kinetics was fitted.RESULTS Salvianolic acid B was stable in acidic and weakly acidic buffer solutions and artificial gastric fluid,which demonstrated poor stability in neutral and alkaline buffer solutions,artificial intestinal fluid,H2O2 and biological matrices.The degradation process of this constituent accorded with the first-order kinetic model in ileum homogenate,and the second-order kinetic model in pH 7.4 buffer solution,artificial intestinal fluid,H2O2 and stomach,duodenum,jejunum,colon homogenates.CONCLUSION Biological matrices,oxidants and alkaline environment can affect the stability of salvianolic acid B.This experimental exhibits important significance for the development and application of salvianolic acid B-related products.

Mutations in the cyclin-dependent kinase-like 5 gene (CDKL5) cause a severe neurodevelopmental disorder, yet the impact of truncating mutations remains unclear. Here, we introduce the Cdkl5^492stop mouse model, mimicking C-terminal truncating mutations in patients. 492stop/Y mice exhibit altered dendritic spine morphology and spontaneous seizure-like behaviors, alongside other behavioral deficits. After creating cell lines with various Cdkl5 truncating mutations, we found that these mutations are regulated by the nonsense-mediated RNA decay pathway. Most truncating mutations result in CDKL5 protein loss, leading to multiple disease phenotypes, and offering new insights into the pathogenesis of CDKL5 disorder.
Animals ; Disease Models, Animal ; Mice ; Protein Serine-Threonine Kinases/deficiency* ; Mutation/genetics* ; Epileptic Syndromes/genetics* ; Humans ; Dendritic Spines/pathology* ; Spasms, Infantile/genetics* ; Male ; Seizures/genetics* ; Mice, Inbred C57BL

This study employed the "disease-medicine-dose" pattern to mine the medication rules of traditional Chinese medicine(TCM) prescriptions containing Astragali Radix in ancient Chinese medical books, aiming to provide a scientific basis for the clinical application of Astragali Radix and the development of new medicines. The TCM prescriptions containing Astragali Radix were retrieved from databases such as Chinese Medical Dictionary and imported into Excel 2020 to construct the prescription library. Statical analysis were performed for the prescriptions regarding the indications, syndromes, medicine use frequency, herb effects, nature and taste, meridian tropism, dosage forms, and dose. SPSS statistics 26.0 and IBM SPSS Modeler 18.0 were used for association rules analysis and cluster analysis. A total of 2 297 prescriptions containing Astragali Radix were collected, involving 233 indications, among which sore and ulcer, consumptive disease, sweating disorder, and apoplexy had high frequency(>25), and their syndromes were mainly Qi and blood deficiency, Qi and blood deficiency, Yin and Yang deficiency, and Qi deficiency and collateral obstruction, respectively. In the prescriptions, 98 medicines were used with the frequency >25 and they mainly included Qi-tonifying medicines and blood-tonifying medicines. Glycyrrhizae Radix et Rhizoma, Angelicae Sinensis Radix, Ginseng Radix et Rhizoma, Atractylodis Macrocephalae Rhizoma, and Citri Reticulatae Pericarpium were frequently used. The medicines with high frequency mainly have warm or cold nature, and sweet, pungent, or bitter taste, with tropism to spleen, lung, heart, liver, and kidney meridians. In the treatment of sore and ulcer, Astragali Radix was mainly used with the dose of 3.73 g and combined with Glycyrrhizae Radix et Rhizoma to promote granulation and heal up sores. In the treatment of consumptive disease, Astragali Radix was mainly used with the dose of 37.30 g and combined with Ginseng Radix et Rhizoma to tonify deficiency and replenish Qi. In the treatment of sweating disorder, Astragali Radix was mainly used with the dose of 3.73 g and combined with Glycyrrhizae Radix et Rhizoma to consolidate exterior and stop sweating. In the treatment of apoplexy, Astragali Radix was mainly used with the dose of 7.46 g and combined with Glycyrrhizae Radix et Rhizoma to dispell wind and stop convulsions. Astragali Radix can be used in the treatment of multiple system diseases, with the effects of tonifying Qi and ascending Yang, consolidating exterior and stopping sweating, and expressing toxin and promoting granulation. According to the manifestations of different diseases, when combined with other medicines, Astragali Radix was endowed with the effects of promoting granulation and healing up sores, tonifying deficiency and Qi, consolidating exterior and stopping sweating, and dispelling wind and replenishing Qi. The findings provide a theoretical reference and a scientific basis for the clinical application of Astragali Radix and the development of new medicines.
Drugs, Chinese Herbal/history* ; Humans ; Medicine, Chinese Traditional/history* ; History, Ancient ; Astragalus Plant/chemistry* ; China ; Astragalus propinquus