Epilepsy affects over 50 million people worldwide. Drug-resistant epilepsy (DRE) accounts for up to a third of these cases, and neuro-inflammation is thought to play a role in such cases. Despite being a long-debated issue in the field of DRE, the mechanisms underlying neuroinflammation have yet to be fully elucidated. The pro-inflammatory microenvironment within the brain tissue of people with DRE has been probed using single-cell multimodal transcriptomics. Evidence suggests that inflammatory cells and pro-inflammatory cytokines in the nervous system can lead to extensive biochemical changes, such as connexin hemichannel excitability and disruption of neurotransmitter homeostasis. The presence of inflammation may give rise to neuronal network abnormalities that suppress endogenous antiepileptic systems. We focus on the role of neuroinflammation and brain network anomalies in DRE from multiple perspectives to identify critical points for clinical application. We hope to provide an insightful overview to advance the quest for better DRE treatments.
Humans ; Drug Resistant Epilepsy/metabolism* ; Neuroinflammatory Diseases/immunology* ; Animals ; Brain/pathology* ; Nerve Net/pathology*

Malocclusion,identified by the World Health Organization(WHO)as one of three major oral diseases,profoundly impacts the dental-maxillofacial functions,facial esthetics,and long-term development of～260 million children in China.Beyond its physical manifestations,malocclusion also significantly influences the psycho-social well-being of these children.Timely intervention in malocclusion can foster an environment conducive to dental-maxillofacial development and substantially decrease the incidence of malocclusion or reduce the severity and complexity of malocclusion in the permanent dentition,by mitigating the negative impact of abnormal environmental influences on the growth.Early orthodontic treatment encompasses accurate identification and treatment of dental and maxillofacial morphological and functional abnormalities during various stages of dental-maxillofacial development,ranging from fetal stages to the early permanent dentition phase.From an economic and societal standpoint,the urgency for effective early orthodontic treatments for malocclusions in childhood cannot be overstated,underlining its profound practical and social importance.This consensus paper discusses the characteristics and the detrimental effects of malocclusion in children,emphasizing critical need for early treatment.It elaborates on corresponding core principles and fundamental approaches in early orthodontics,proposing comprehensive guidance for preventive and interceptive orthodontic treatment,serving as a reference for clinicians engaged in early orthodontic treatment.

Objective:To explore the application value of histogram and texture feature analysis based on CT images in distinguishing long bone osteosarcoma (OS) from Ewing sarcoma (ES).Methods:A retrospective collection of 25 patients with long bone osteosarcoma and 25 patients with Ewing sarcoma confirmed by surgery and pathology in National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Qilu Hospital of Shandong University and Nanjing Drum Tower Hospital, Nanjing University Medical School, from March 2018 to May 2023 was conducted. All patients were randomly divided into a training set (21 cases of OS and 19 cases of ES) and a validation set (4 cases of OS and 6 cases of ES) in an 8∶2 ratio. The region of interest (ROI) on CT images to extract texture feature parameters was manually sketched. Random forest and least absolute shrinkage and selection operator (LASSO) algorithm were used for feature screening. Logistic regression (LR), random forest (RF), support vector machine (SVM) and K-nearest neighbor (KNN) classifiers were used to establish models respectively. Receiver operating characteristic (ROC)curve was drawn and area under the curve (AUC) was calculated to evaluate the diagnostic efficiency of the four models.Results:A total of 100 texture parameters were extracted from CT images, and 8 feature parameters (maximum 3D diameter, 10th percentile, kurtosis, maximum pixel intensity value, inverse normalization, grayscale level variance, long range high grayscale emphasis, and low grayscale area emphasis) were obtained through screening. Four classifiers were used to establish models, and the AUC values of the four models (LR, RF, SVM, KNN) in the validation group were 0.92, 0.79, 0.83, and 0.73, respectively. LR and SVM classifier algorithm trains models had high diagnostic efficiency, with an accuracy of 90%, sensitivity of 83%, specificity of 100%, and AUC of 92% for the LR classifier validation set; the accuracy of SVM classifier validation set was 80%, sensitivity was 67%, specificity was 100%, and AUC was 83%.Conclusions:LR and SVM models have high value in distinguishing OS and ES.

OBJECTIVE: To evaluate the surgical methods for treating distal urethral stricture. METHODS: The clinical data of 80 patients with distal urethral stricture in Beijing Jishuitan Hospital, Captial Medical University between January 2018 and December 2022 were retrospectively collected. Including male genital lichen sclerosus (MGLS) 33 cases, iatrogenic injury 25 cases, postoperative hypospadias 12 cases, and other causes such as trauma 10 cases. Among these cases, strictures involved the urethral meatus in 54 instances, of which 38 were treated with meatotomy (MO), 7 with penile skin flap urethroplasty (PSFU), and 9 with oral mucosa graft urethroplasty (OMGU). There were also 26 instances where strictures involved both the navicular fossa and meatus; one case underwent PSFU while 25 underwent OMGU. Based on different surgical methods used for treatment purposes we divided the patients into MO group, PSFU group and OMGU group. The age of the three groups was (48.8±20.0) years, (53.3±21.8) years and (44.5±16.4) years. The mean±SD body mass index (BMI) was (28.6±3.9) kg/m², (29.6±3.2) kg/m² and (29.2±4.8) kg/m². The preoperative maximum flow rate was (5.8±2.3) mL/s, (6.8±2.4) mL/s and (5.7±3.1) mL/s. RESULTS: All the operations were successfully completed without Clavien Ⅲ or Ⅳ complications. The median length of strictures (measured intraoperatively) in the three groups were 1.1 (1.0, 1.6), 1.5 (1.1, 2.0) and 4.0 (2.5, 5.0) cm. The median operation time was 60.0 (60.0, 75.0), 85.0 (75.0, 112.5) and 180.0 (75.0, 330.0) min. The median estimated blood loss was 5.0 (2.0, 10.0), 15.0 (5.0, 42.5) and 180.0 (135.0, 216.3) mL. The median postoperative hospital stay was 3.5 (2.0, 5.0), 6.5 (3.5, 7.0) and 7.5 (7.0, 11.3) days. The median follow-up duration was 40.0 (26.3, 57.3), 55.0 (18.8, 62.8) and 52.5 (30.5, 64.0) months. The median postoperative maximum flow rate was 18.3 (15.5, 19.8), 19.2 (16.1, 20.1) and 17.2 (14.2, 19.6) mL/s. Among the 38 patients with MO, 33 cases had normal urination without reintervention, and 5 cases experienced recurrent strictures and required regular urethral dilation. Among the 8 patients with PSFU, 7 cases had normal urination without reintervention, and one case developed a urinary fistula, for which intervention was recommended but the patient opted to maintain the status quo. Among the 34 patients with OMGU, 28 cases had normal urination without reintervention. There were 6 instances of stenosis recurrence, with 5 cases requiring regular urethral dilations and one case requiring reconstructive surgery. The overall success rate of operation was 85.0%, and the reintervention rate was 15.0%. Statistical analysis revealed significant differences in etiologies among the three groups (P=0.002), as well as in stricture locations (P < 0.001), length of strictures (P < 0.001), operation time (P < 0.001), estimated blood loss (P < 0.001) and postoperative hospital stays (P < 0.001). However, no significant differences were observed in terms of age, BMI, history of previous urethral stricture surgeries, preoperative maximum flow rate, follow-up duration, postoperative maximum flow rate and reintervention rate. Univariate and multivariate Logistic regression analyses indicated that a history of previous urethral stricture surgeries was a risk factor for postoperative reintervention (P=0.026). CONCLUSION MO and PSFU are primarily suitable for treating short-segment (≤1.5 cm) distal penile urethral strictures, whereas OMGU is more appropriate for longer segment strictures. MO and OMGU can both be utilized in managing MGLS cases. PSFU and OMGU are more conducive to improving the appearance of the urethral meatus. The success rate of surgical management of distal penile urethral stricture is 85.0%, 15.0% of the patients still require surgical intervention after surgery, and having history of previous urethral stricture surgeries is a risk factor for postoperative reintervention.
Humans ; Urethral Stricture/surgery* ; Male ; Retrospective Studies ; Middle Aged ; Adult ; Urethra/surgery* ; Surgical Flaps ; Penis/surgery* ; Mouth Mucosa ; Urologic Surgical Procedures, Male/methods* ; Plastic Surgery Procedures/methods* ; Lichen Sclerosus et Atrophicus/surgery* ; Operative Time ; Skin Transplantation/methods* ; Aged

To preliminarily understand the pathogenic mechanism of Mycoplasma genitalium (Mg) GroEL protein, we used bioinformatics tools to predict the structure and function of Mg GroEL protein and then constructed the recombinant plasmid pET-28a-GroEL. The protein expression was induced by 0.2 mmol/L IPTG, and the expressed protein was purified by Ni-iminodicitic acid (IDA) column affinity. Tohoku Hospital Pediatrics-1 (THP-1) cells were exposed to 2 μg/mL Mg rGroEL. The levels of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in the cell supernatant were measured by ELISA, and that of IL-6 was measured by an automatic chemiluminescence instrument. The activation of the nuclear factor-kappa B (NF-κB) signaling pathway was visualized by immunofluorescence and Western blotting. The results showed that Mg GroEL was a stable hydrophilic protein composed of 543 amino acid residues, with the relative molecular mass of 58.44 kDa, an isoelectric point of 5.68, and a molecular formula of C₂₅₆₈H₄₃₀₀N₇₀₀O₈₂₅S₈. The secondary structure was mainly composed of α-helices and random coils. Mg GroEL contained 12 B-cell dominant epitopes and 10 T-cell dominant epitopes. It exhibited high homology with the GroEL proteins from Mycoplasma pneumoniae, M. agalactiae, M. arthritidis, M. hyopneumoniae, and M. bovis. Mg rGroEL activated the NF-κB signaling pathway and promoted the secretion of IL-1β, IL-6, and TNF-α in THP-1 cells. These results suggest that Mg GroEL exhibits substantial antigenicity and possesses the capability of triggering inflammation in host cells. This study establishes a theoretical basis for future investigations pertaining to the role and pathogenic mechanisms of Mg GroEL.
Mycoplasma genitalium/metabolism* ; Chaperonin 60/metabolism* ; Computational Biology ; Bacterial Proteins/genetics* ; Humans ; NF-kappa B/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Interleukin-1beta/genetics* ; Inflammation ; Interleukin-6/genetics* ; Recombinant Proteins/genetics* ; THP-1 Cells ; Signal Transduction ; Escherichia coli/metabolism*

Objective:To investigate the appropriate pretreatment methods for single cell RNA sequencing of airway aspirate cells.Methods:Four fresh airway aspirate specimens were collected from four patients with acute respiratory tract infections. These specimens were digested with airway aspirate digester and prepared into single cell suspension. The cells were used for library construction directly (DE), or fixed with 10×Genomics Chromium Next GEM Single Cell Fixed RNA Sample Preparation Kit and then mixed to construct the library (DF), or cryopreserved, thawed, fixed (FF) before mixed to construct the library. All three methods were treated with oil emulsion using 10 4 cells and subjected to single-cell sequencing using the 10×Genomics platform. The number of obtained cells, data quality, annotated cell types and expression of marker genes were analyzed. Differences in the expression of highly variable genes (HVGs) of the same cell subsets obtained by the three pretreatment methods were compared using Pearson correlation. Expression of the differentially expressed genes in the same cell subpopulation obtained by different pretreatment methods was also compared. The correlation of the expression of differentially expressed genes between the same cell subsets obtained by the three pretreatment methods was analyzed by Pearson correlation. Results:The median numbers of single cells obtained using DE, FF and DF methods were 2 733, 1 140 and 5 897 ( P>0.05). The unique molecular identifiers were higher than 500. The median numbers of genes obtained using the three methods were 801, 887 and 1 259 ( P>0.05). The cells with novelty score over 0.8 accounted for 99%, 87% and 93%, respectively. There were nine cell subsets obtained by the three methods, including squamous cells, secretory cells, ciliated cells, T cells, B cells, macrophages, plasma cells and neutrophils. DF and FF methods could obtain more basal cells with specific high expression of keratin 5 than DE method. The differentially expressed and highly variable genes in the same cell subsets obtained by the three pretreatment methods showed high consistency in their expression with a significant correlation ( P<0.001). Conclusions:Under the same sequencing data volume, the quality of data obtained from fixed airway aspirate single-cell suspensions using the method of probe hybridization and transcriptome sequencing was comparable to that obtained directly from fresh cells. This method was more suitable for the pretreatment of clinical samples used for single-cell RNA sequencing.

Articular cartilage (AC) injuries often lead to cartilage degeneration and may ultimately result in osteoarthritis (OA) due to the limited self-repair ability. To date, numerous intra-articular delivery systems carrying various therapeutic agents have been developed to improve therapeutic localization and retention, optimize controlled drug release profiles and target different pathological processes. Due to the complex and multifactorial characteristics of cartilage injury pathology and heterogeneity of the cartilage structure deposited within a dense matrix, delivery systems loaded with a single therapeutic agent are hindered from reaching multiple targets in a spatiotemporal matched manner and thus fail to mimic the natural processes of biosynthesis, compromising the goal of full cartilage regeneration. Emerging evidence highlights the importance of sequential delivery strategies targeting multiple pathological processes. In this review, we first summarize the current status and progress achieved in single-drug delivery strategies for the treatment of AC diseases. Subsequently, we focus mainly on advances in multiple drug delivery applications, including sequential release formulations targeting various pathological processes, synergistic targeting of the same pathological process, the spatial distribution in multiple tissues, and heterogeneous regeneration. We hope that this review will inspire the rational design of intra-articular drug delivery systems (DDSs) in the future.

Objective:To explore the innovation mode of independent transformation-oriented science and technology research program approval by medical new research & development (R&D) institution.Methods:Through analyzing the program layout, funds, review experts, undertaking units, chief experts and interdisciplinarity to summarize the experiences of the independent transformation-oriented municipal program approval by Haihe laboratory of Cell Ecosystem in 2022.Results:As a new medical R&D institution, which vigorously constructed by Tianjin, Haihe laboratory of Cell Ecosystem has carried out the practice of the independent transformation-oriented municipal program through the measures of layout of full-chain transformation, conducting transformation-oriented review, gathering high-level research talents, and emphasizing interdisciplinarity.Conclusions:The experiences of Haihe laboratory of Cell Ecosystem make significance for medical new R&D institutions to explore and cultivate scientific research program with transformation potential and to promote the transformation of scientific and technological achievements, which are powerful factors for new R&D institutions to play a role of pilot and provide important support to scientific and technological innovation and transformation.