ObjectiveCell adhesion is a critical process that regulates cellular physiological functions. Quantitative characterization of adhesion dynamics is essential for elucidating the intrinsic mechanical mechanisms underlying cellular activities. Although atomic force microscopy-based single-cell force spectroscopy is widely used for single-cell adhesion measurements, it requires complex chemical modifications for preparation of live-cell probes, leading to limitations such as cumbersome operation, low throughput, and potential impacts on cell viability. Fluidic force microscopy, which combines atomic force microscopy with microfluidic probes, is a technique allowing the operation of force-controlled nanopipettes in aqueous environments. By applying negative or positive pressure via a pressure controller, a single living cell can be captured onto or released from the cantilever under physiological conditions. This procedure offers a simple workflow and high assay throughput for single-cell adhesion measurements without the need for chemical functionalization. In this study, fluidic force microscopy-based single-cell force spectroscopy was adopted to achieve long-term quantitative characterization of single-cell adhesion dynamics in a simpler and more efficient manner, comparing the dynamic differences in adhesion establishment between two cell lines with different differentiation levels. MethodsHEK 293T and hTERT RPE-1 cells were non-invasively captured on the cantilever of a fluidic force microscope via its integrated microfluidic system during 40 h of adhesion culture. Cell-substrate detachment assays were performed, and force-distance curves were recorded to extract key mechanical adhesion parameters, including adhesion force, adhesion energy, and maximum detachment distance. These measurements were combined with real-time monitoring of cell spreading area to systematically characterize the dynamic evolution of single-cell adhesion. ResultshTERT RPE-1 cells rapidly entered a stable adhesion phase within 1 h after seeding, with both area-normalized adhesion force and area-normalized adhesion energy reaching peak values. In contrast, HEK 293T cells required 4 h to achieve stable adhesion. Subsequently, the adhesion force, adhesion energy and maximum detachment distance of hTERT RPE-1 and HEK 293T cells stabilized at approximately 240 nN vs. 30 nN, 2.2 pJ vs. 0.12 pJ and 6 μm vs. 4 μm, respectively. hTERT RPE-1 cells reached the peak of area-normalized adhesion parameters earlier than HEK 293T cells, with their peak area-normalized adhesion force and area-normalized adhesion energy being substantially elevated relative to HEK 293T cells. HEK 293T cells presented stronger linear correlations among adhesion energy, maximum detachment distance and adhesion force compared with hTERT RPE-1 cells. For both cell lines, cell spreading area exhibited a weak correlation with adhesion force. Whereas the area-normalized adhesion parameters of HEK 293T cells remained relatively constant throughout the adhesion process, hTERT RPE-1 cells exhibited elevated values in the early phase, followed by a gradual decline. These results indicated distinct dynamic adhesion patterns between the two cell types, with hTERT RPE-1 cells exhibiting stronger adhesion strength and higher adhesion efficiency. ConclusionIn this study, fluidic force microscopy-based single-cell force spectroscopy was successfully applied to perform long-term in situ quantitative measurement of the adhesion dynamics in single adherent cells. The approach revealed divergent adhesion patterns between HEK 293T and hTERT RPE-1 cells, suggesting a close association between cell differentiation and adhesion behaviors. These findings provide quantitative mechanical evidence for further understanding the underlying mechanisms of cell adhesion.

Epilepsy is a chronic neurological disorder affecting ~65 million individuals worldwide. Abnormal synaptic plasticity is one of the most important pathological features of this condition. We investigated how ubiquitin-specific peptidase 47 (USP47) influences synaptic plasticity and its link to epilepsy. We found that USP47 enhanced excitatory postsynaptic transmission and increased the density of total dendritic spines and the proportion of mature dendritic spines. Furthermore, USP47 inhibited the degradation of the ubiquitinated α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) subunit glutamate receptor 1 (GluR1), which is associated with synaptic plasticity. In addition, elevated levels of USP47 were found in epileptic mice, and USP47 knockdown reduced the frequency and duration of seizure-like events and alleviated epileptic seizures. To summarize, we present a new mechanism whereby USP47 regulates excitatory postsynaptic plasticity through the inhibition of ubiquitinated GluR1 degradation. Modulating USP47 may offer a potential approach for controlling seizures and modifying disease progression in future therapeutic strategies.
Animals ; Receptors, AMPA/metabolism* ; Neuronal Plasticity/physiology* ; Seizures/physiopathology* ; Disease Models, Animal ; Mice, Inbred C57BL ; Mice ; Ubiquitin Thiolesterase/genetics* ; Male ; Excitatory Postsynaptic Potentials/physiology* ; Ubiquitination ; Dendritic Spines/metabolism* ; Hippocampus/metabolism*

OBJECTIVES: To study the impact of different induction remission therapies on the nutritional status of children with Crohn's disease (CD). METHODS: A retrospective analysis was conducted on the medical data of 188 children with CD who received induction remission therapy at the Children's Hospital, Zhejiang University School of Medicine from January 2013 to May 2023. The effects of three induction remission therapies-exclusive enteral nutrition (EEN), corticosteroids, and infliximab (IFX)-on height, weight, hemoglobin, and trace elements were compared. RESULTS: After induction remission therapy, the weight-for-age Z scores and serum levels of hemoglobin and albumin in all three groups significantly increased (P<0.05), while the height-for-age Z scores showed no change (P>0.05). The levels of folic acid, 25-hydroxy vitamin D, serum iron, and total calcium increased, and serum copper levels decreased in the EEN and IFX groups (P<0.05). Vitamin B₁₂ levels improved in the EEN group (P<0.05). CONCLUSIONS EEN, corticosteroids, and IFX may effectively improve the weight-for-age Z scores and serum levels of hemoglobin and albumin in children with CD. EEN and IFX seem to be superior to corticosteroid therapy in improving 25-hydroxy vitamin D, folic acid, serum iron, and total calcium levels.
Humans ; Crohn Disease/blood* ; Child ; Male ; Female ; Retrospective Studies ; Adolescent ; Nutritional Status ; Infliximab/therapeutic use* ; Child, Preschool ; Enteral Nutrition ; Adrenal Cortex Hormones/therapeutic use*

OBJECTIVES: To evaluate the effectiveness of single-balloon and double-balloon enteroscopy in diagnosing pediatric small bowel diseases and assess the diagnostic efficacy of computed tomography enterography (CTE) for small bowel diseases using enteroscopy as the reference standard. METHODS: Clinical data from 576 children who underwent enteroscopy at Hunan Children's Hospital between January 2017 and December 2023 were retrospectively collected. The children were categorized based on enteroscopy type into the single-balloon enteroscopy (SBE) group (n=457) and double-balloon enteroscopy (DBE) group (n=119), and the clinical data were compared between the two groups. The sensitivity and specificity of CTE for diagnosing small bowel diseases were evaluated using enteroscopy results as the standard. RESULTS: Among the 576 children, small bowel lesions were detected by enteroscopy in 274 children (47.6%).There was no significant difference in lesion detection rates or complication rates between the SBE and DBE groups (P>0.05), but the DBE group had deeper insertion, longer procedure time, and higher complete small bowel examination rate (P<0.05). The complication rate during enteroscopy was 4.3% (25/576), with 18 cases (3.1%) of mild complications and 7 cases (1.2%) of severe complications, which improved with symptomatic treatment, surgical, or endoscopic intervention. Among the 412 children who underwent CTE, the sensitivity and specificity for diagnosing small bowel diseases were 44.4% and 71.3%, respectively. CONCLUSIONS SBE and DBE have similar diagnostic efficacy for pediatric small bowel diseases, but DBE is preferred for suspected deep small bowel lesions and comprehensive small bowel examination. Enteroscopy in children demonstrates relatively good overall safety. CTE demonstrates relatively low sensitivity but comparatively high specificity for diagnosing small bowel diseases.
Retrospective Studies ; Treatment Outcome ; Double-Balloon Enteroscopy/statistics & numerical data* ; Single-Balloon Enteroscopy/statistics & numerical data* ; Humans ; Male ; Female ; Child ; Operative Time ; Tomography, X-Ray Computed/statistics & numerical data* ; Sensitivity and Specificity ; Intestine, Small/surgery* ; Intestinal Diseases/surgery*

OBJECTIVE: This study aimed to evaluate the association between susceptibility genes and non-alcoholic fatty liver disease (NAFLD) in children with obesity. METHODS: We conducted a two-step case-control study. Ninety-three participants were subjected to whole-exome sequencing (exploratory set). Differential genes identified in the small sample were validated in 1,022 participants using multiplex polymerase chain reaction and high-throughput sequencing (validation set). RESULTS: In the exploratory set, 14 genes from the NAFLD-associated pathways were identified. In the validation set, after adjusting for sex, age, and body mass index, ECI2 rs2326408 (dominant model: OR = 1.33, 95% CI: 1.02-1.72; additive model: OR = 1.22, 95% CI: 1.01-1.47), C6orf201 rs659305 (dominant model: OR = 1.30, 95% CI: 1.01-1.69; additive model: OR = 1.21, 95% CI: 1.00-1.45), CALML5 rs10904516 (pre-ad dominant model: OR = 1.36, 95% CI: 1.01-1.83; adjusted dominant model: OR = 1.40, 95% CI: 1.03-1.91; and pre-ad additive model: OR = 1.26, 95% CI: 1.04-1.66) polymorphisms were significantly associated with NAFLD in children with obesity ( P < 0.05). Interaction analysis revealed that the gene-gene interaction model of CALML5 rs10904516, COX11 rs17209882, and SCD5 rs3733228 was optional ( P < 0.05), demonstrating a negative interaction between the three genes. CONCLUSION In the Chinese population, the CALML5 rs10904516, C6orf201 rs659305, and ECI2 rs2326408 variants could be genetic markers for NAFLD susceptibility.
Humans ; Non-alcoholic Fatty Liver Disease/genetics* ; Child ; Male ; Female ; Genetic Predisposition to Disease ; Case-Control Studies ; Exome Sequencing ; Adolescent ; Polymorphism, Single Nucleotide ; Obesity/complications* ; Pediatric Obesity/complications* ; China

Sugarcane (Saccharum spp.) is an important sugar crop. Biotic and abiotic stresses such as diseases, cold, and drought are major factors limiting sugarcane production. β-1,3-glucanase (EC 3.2.1.39), a member of the pathogenesis-related protein family, plays an essential role not only in the plant defenses against pathogens but also in plant growth, development, and abiotic stress responses. To systematically investigate the sugarcane β-1,3-glucanase gene family, 132 glycoside hydrolase (GH) 17 family members were identified in the genomes of the sugarcane wild species Saccharum spontaneum 'Np-X', the tropical species S. officinarum 'LA-Purple', and the Saccharum spp. hybrid cultivar 'R570'. The results of the phylogenetic analysis categorized them into four subfamilies, of which subfamily Ⅳ had the largest proportion of members (102). The members of the sugarcane GH17 gene family contained five conserved motifs and 0-16 introns. The majority of the GH17 genes exhibited a genome-wide replication pattern, with 89.50% originating from S. spontaneum 'Np-X' and S. officinarum 'LA-Purple', while 58.10% of them in the Saccharum spp. hybrid cultivar 'R570' belonged to the discrete replication type. Four major classes of cis-acting elements were identified in the promoters, including the elements related to plant growth, development, and tissue-specific expression (14.21%), light-responsive elements (38.24%), biotic or abiotic stress-responsive elements (9.18%), and hormone-responsive elements (38.37%), which suggested that this gene family was involved in plant growth, development, hormone responses, and stress responses. Transcriptome and quantitative real-time PCR (RT-qPCR) analyses showed that the sugarcane GH17 genes exhibited tissue-specific expression and were differentially expressed under low temperature, drought, and hormone treatments, as well as during the interactions between different sugarcane genotypes and Sporisorium scitamineum, suggesting their potential roles in plant defenses. In addition, some SsGlu genes (SsGlu5, SsGlu20, SsGlu21, SsGlu25, SsGlu28, and SsGlu39) were expected to serve as candidate stress-related genes. This study lays a foundation for further revealing the molecular mechanisms of the stress resistance of sugarcane via β-1,3-glucanase genes.
Saccharum/physiology* ; Stress, Physiological/genetics* ; Glucan 1,3-beta-Glucosidase/metabolism* ; Multigene Family ; Phylogeny ; Gene Expression Regulation, Plant ; Plant Proteins/genetics*

OBJECTIVE: To investigate effectiveness of simplified all-arthroscopic Broström technique in treatment of chronic lateral ankle instability in adolescents. METHODS: A clinical data of 21 adolescent patients with chronic lateral ankle instability, who met the selection criteria and were admitted between June 2023 and May 2024, was retrospectively analyzed. There were 18 males and 3 females with an average age of 16.0 years (range, 13-18 years). There were 9 cases of left ankle joint injury and 12 cases of right ankle joint injury. Anterior talofibular ligament (ATFL) injury was diagnosed by arthroscopy in all patients. There were 11 cases of cartilage injury, 5 cases of avulsion fractures, and 6 cases of ankle impingement syndrome. The time from first sprain to operation ranged from 3-60 months (mean, 12.0 months). The ATFL was repaired and the ankle joint stability was restored by simplified all-arthroscopic Broström technique. Visual analogue scale (VAS) score, Tegner score, American Orthopaedic Foot and Ankle Society (AOFAS) score, Karlsson ankle function scale (KAFS) score, Foot and Ankle Outcome Score (FAOS) were used to evaluate ankle pain and function. MRI was used to evaluate the ligament healing. RESULTS: All patients were followed up 8-15 months (mean, 12.6 months). After operation, 1 patient suffered from superficial peroneal nerve injury, 1 patient developed anterior scar impingement on the ankle, 2 patients had superficial wound infection, and 1 patient suffered from sprain again. The VAS score, Tenger score, AOFAS score, KAFS score, and FAOS score significantly improved when compared with the preoperative scores ( P<0.05). MRI examination showed the ligament healing and good tension. CONCLUSION For adolescent patients with chronic lateral ankle instability, using simplified all-arthroscopic Broström technique to repair ATFL can effectively alleviate ankle pain, improve stability, and achieve good effectiveness.
Humans ; Joint Instability/surgery* ; Male ; Adolescent ; Female ; Arthroscopy/methods* ; Ankle Joint/physiopathology* ; Retrospective Studies ; Ankle Injuries/surgery* ; Lateral Ligament, Ankle/injuries* ; Treatment Outcome ; Chronic Disease ; Range of Motion, Articular

Objective: To understand the prevalence and related factors of depressive symptoms among primary and secondary school students in the in depth monitoring counties of China s Rural Compulsory Education Nutrition Improvement Program, so as to provide a basis for prevention and psychological intervention of depressive symptoms among children and adolescents in rural areas. Methods: In November 2022, a stratified random sampling method was adopted to collect height and weight data, basic personal and family information of 7 949 primary and secondary school students from grade three to grade nine through physical measurements and questionnaires in 56 key monitoring schools implementing the Student Nutrition Improvement Program in 7 in depth monitoring counties (Jalaid Banner in Inner Mongolia, Jinzhai County in Anhui, Mao Xian in Sichuan, Tiandeng County in Guangxi, Mian County in Shaanxi, Zhaozhou County in Heilongjiang and Youxi County in Fujian), and to obtain the information related to their depressive symptoms through the self assessment questionnaire on depression. Multivariate Logistic regression analysis was conducted to analyze the prevalence of depressive symptoms among primary and secondary school students, as well as their related factors. Results: The detection rate of depressive symptoms among primary and secondary school students in the in depth monitored counties was 23.5%. Logistic regression analysis showed that the probability of detecting depressive symptoms was higher among female students, middle school students, students whose video screen duration per day was >2 h, and students whose parents marital status was divorced or widowed ( OR =1.40, 1.64, 1.60, 1.24), and students whose sleep duration reached the recommended standard, whose parents usually accompanied them daily for time was 60-<120 min and ≥120 min, and students whose mothers literacy level was middle school graduation had lower probability of detecting depressive symptoms ( OR =0.85, 0.84, 0.71, 0.76) ( P < 0.05 ). Conclusion The detection rate of depressive symptoms among students in the in depth monitoring area is high, and targeted interventions need to be developed for students to reduce the risk of mental health problems.

Objective:This study aims to explore the clinical characteristics of patients with multiple myeloma concurrently presenting with a second primary malignancy.By analyzing the causes based on literature reports,this study aims to deepen the understanding of multiple primary cancers in multiple myeloma patients,and provide some assistance for auxiliary examinations and risk assessments of multiple myeloma patients.Methods:A retrospective analysis was conducted on the clinical data of four patients with multiple myeloma who also had a second primary malignancy admitted to The Second Affiliated Hospital,Chongqing Medical University.Each patient was diagnosed with multiple myeloma through bone marrow aspiration and had a second primary malignancy confirmed through pathological biopsy.Results:The onset age of the 4 patients ranged from 42 to 81 years old;three were males and one was female.One patient had smoldering myeloma with a second primary hematological malignancy(follicular lymphoma),while the other 3 patients had concurrent solid malignancies,including bladder cancer,esophageal cancer,and breast cancer,respectively.Conclusion:Multiple myeloma accompanied by a second primary malignancy is rare,and early pathological biopsy is necessary for diagnosis to avoid missed diagnosis.

Objective:To evaluate the effectiveness and safety of belumosudil for the treatment of chronic graft-versus-host disease (cGVHD) .Methods:We retrospectively collected data on patients with cGVHD who received belumosudil at Ruijin Hospital Affiliated to Shanghai Jiaotong University School of Medicine from May 2023 to March 2024. The study endpoints were overall response rate (ORR), organ-specific response rates, time to response (TTR), changes in Lee Symptom Scale (LSS) scores, tapering or discontinuation of corticosteroid treatment, failure-free survival (FFS), and adverse events.Results:The study included 20 patients with cGVHD who received belumosudil, of whom 15 were men and 5 women. The median age was 34.5 (12-67) years, and three patients were under 18 years old. The median follow-up duration was 5.0 (1.4 - 9.8) months. All patients had severe cGVHD, and 18 (90.0%) showed involvement of at least four organs. The median number of prior treatment lines was 4, and 15 patients (75%) had previously received ruxolitinib. All patients received 200 mg of belumosudil once daily in combination with other cGVHD systemic therapies. The ORR was 90.0% (95% CI: 68.3%-98.8%), and all responses were partial responses. The median TTR was 1.6 (0.9 - 8.4) months. The LSS scores improved in a clinically meaningful way in 80.0% (16/20) of the patients within 3 months. The corticosteroid dose was reduced in 42.6% (6/14) of the patients. The 3-month FFS was 79.6% (95% CI: 61.4%-100.0%). Most adverse events were grade 1 or grade 2, and two patients (10.0%) experienced grade 3 or higher-grade adverse events. Conclusions:In the real-world setting, belumosudil demonstrated good effectiveness and safety in patients with cGVHD with a history of severe disease and multiorgan involvement.