In recent years， repetitive transcranial magnetic stimulation （rTMS） has garnered significant attention as a therapeutic approach for sleep disorders in the elderly. However， the prevailing rTMS protocols are predominantly developed based on normative neurophysiological data derived from young adults and fail to incorporate individualized parameters tailored to the brain characteristics of the elderly. To address this gap， the consensus development group synthesized the latest evidence from 2010 to 2025 and established a standardized rTMS protocol specifically for elderly patients with sleep disorders. Adhering to the Appraisal of Guidelines for Research and Evaluation II （AGREE II） framework， systematically screened randomized controlled trials （RCTs） and systematic reviews regarding rTMS in the treatment of sleep disorders across various conditions. Meanwhile， the Grading of Recommendations Assessment， Development and Evaluation （GRADE） system was employed to rigorously grade the quality of evidence and the strength of recommendations. This consensus guideline delineates precise rTMS protocols for the management of sleep disorders in the elderly， highlights the adjustment of stimulation intensity according to scalp-cortex distance recommends either MRI‑guided neuronavigation or the Beam F3/F4 heuristic approach for accurate target localization， thereby providing precise rTMS intervention protocol for sleep disorders in the elderly， aiming to enhance clinical efficacy while ensuring treatment safety. ［Funded by National Key Research and Development Program （number， 2023YFC3603200）； General Program of Shenzhen Science and Technology Innovation Commission （number， JCYJ20240813112859008， JCYJ20240813112900002）； Youth Program of Shenzhen Kangning Hospital （number， KN2023A004）； www.guidelines-registry.cn number， PREPARE-2026CN530］

Bladder tumor is a common tumor in the urinary system, posing a serious threat to patients’ health and quality of life. The standard surgical method for non-muscle-invasive bladder cancer is the transurethral resection of bladder tumor. As a new type of minimally invasive treatment, transurethral holmium laser has shown unique advantages in the treatment of bladder tumors. This work introduces the epidemiological characteristics of bladder tumors and the limitations of traditional treatment methods and elaborates on the technical principles, development history of holmium laser and its application status in the treatment of bladder tumors. By analyzing clinical research data, it compares therapeutic effects between holmium laser and other treatment methods. It also presents the key points of surgical techniques, perioperative management strategies, and measures for the prevention and treatment of complications. This research discusses the current technical challenges and future development directions in holmium laser treatment, providing references for clinical practice and research.

Background: and Purpose Bilirubin has potent antioxidant, anti-inflammatory, and neuroprotective effects. Herein, we investigated whether total bilirubin (TBIL) modifies the association between hypertension and cerebral small vessel disease (CSVD). Methods: Data were obtained from the PolyvasculaR Evaluation for Cognitive Impairment and vaScular Events study. TBIL and direct bilirubin (DBIL) levels were assayed using fasting venous blood samples. Indirect bilirubin (IBIL) was calculated by subtracting DBIL from TBIL. TBIL was stratified as ≤17 μmol/L and >17 μmol/L based on the biological relevance of Gilbert’s syndrome. Hypertension was defined as blood pressure ≥140/90 mm Hg, self-reported hypertension history, or current use of antihypertensive agents. White matter hyperintensity, lacunes, cerebral microbleeds, and enlarged perivascular spaces were evaluated using magnetic resonance imaging and used to rate CSVD burden according to the criteria proposed by Wardlaw et al. and Rothwell et al. Results: This study included 3,061 participants, with a mean age of 61.2±6.7 years and 46.5% males. After adjusting for confounders, hypertension was associated with increased odds of presence of CSVD (Wardlaw: odds ratio [OR]=1.86, 95% confidence interval [CI] 1.41–2.44, P<0.001; Rothwell: OR=1.84, 95% CI 1.43–2.38, P<0.001) and higher modified total CSVD burden (common OR: 1.85, 95% CI 1.45–2.36, P<0.001) in participants with TBIL ≤17 μmol/L but not in TBIL >17 μmol/L (P for interaction <0.05). Johnson–Neyman analyses showed cut-off concentrations of 22.3–22.4 μmol/L for effect modification by TBIL. IBIL contributed to effect modification, whereas DBIL did not. Conclusions Mildly elevated TBIL may modify the association between hypertension and CSVD.

Background/Aims: Cholangiocarcinoma (CCA) is a primary malignant neoplasm with an extremely poor prognosis. While combined chemoradiotherapy has been demonstrated to delay CCA progression to a certain extent, the absence of specific molecular biomarkers or targets significantly hinders the diagnosis and treatment of CCA. Methods: Through cross-analysis of proteomics and ADMA modificationomics, we identified DDX1 overexpressed in CCA with elevated R602-ADMA modifications. HPLC-MS/MS identified PRMT1 as the methyltransferase and USP10 as the deubiquitinating enzyme for DDX1. Immunofluorescence and nuclear-cytoplasmic partitioning experiments confirmed DDX1’s nuclear localization. GO and KEGG analyses clarify the biological functions of DDX1 in response to hypoxia. RNA-seq transcriptomics analyzed key pathways influenced by DDX1. A hydrodynamic in situ CCA mouse model was established to validate the chemopreventive effects of the PRMT1-specific inhibitor GSK715 on CCA development. Results: DDX1 promotes CCA progression both in vivo and in vitro and can be inhibited by GSK715. Mechanistically, PRMT1 mediates ADMA modification at position R602 of DDX1. This modification promotes DDX1 nuclear localization by recruiting USP10 to deubiquitinate DDX1, while simultaneously inhibiting PRMT1 degradation. DDX1 promotes the transcription of PRMT1 and USP10 by binding to the mRNA 3’UTR region, establishing a positive feedback regulatory pathway. This mechanism promotes the occurrence and development of CCA and can serve as a target for the inhibitor GSK715 to suppress CCA progression. Conclusions Our study identified DDX1-R602-ADMA modification as a novel ADMA modification in CCA. It further confirmed its pivotal role in CCA progression. Targeting the USP10-PRMT1-DDX1 axis may represent a significant therapeutic approach for CCA.

ObjectiveTo summarize the application experience of the "Three-Separation" model in the pre-service training of laboratory animal practitioners and scientifically evaluate its training effectiveness. MethodsA retrospective analysis was conducted on 8 009 practitioners who participated in pre-service training at the Fujian Provincial Laboratory Animal Training Base from January 2010 to December 2025. Among them, 1 250 trainees from January 2010 to December 2017 were included in the control group (traditional training model), and 6 759 trainees from January 2018 to December 2025 were included in the evaluation group ("Three-Separation" model, based on separating job requirements, differentiating exam content, and categorizing exam methods). Meanwhile, a questionnaire survey was conducted to evaluate training effectiveness, yielding 1 624 valid questionnaires (585 from the control group and 1 039 from the evaluation group). The "Three-Separation" model simultaneously optimized fee structures to improve affordability and inclusiveness. The comprehensive competencies and training satisfaction of the two groups after training were compared and analyzed using the independent sample t-test, χ² test, and Wilcoxon rank-sum test. ResultsThe overall satisfaction rate of the evaluation group was 92.49% (961/1 039), significantly higher than that of the control group (77.78%, 455/585), with a highly significant difference (χ²=72.50, P<0.001). The evaluation group showed a more favorable distribution of positive, neutral, and negative ratings in dimensions such as course pertinence, time flexibility, assessment rationality, and reasonableness of fees, and the positive rating rates were highly significantly higher than those in the control group (P<0.001). The comprehensive pass rate of the evaluation group (95.53%) was significantly higher than that of the control group (86.24%), with a highly significant difference (P<0.001). ConclusionThe scientific application of the "Three-Separation" model can accurately match the competency requirements of different positions, optimize the training and assessment system, reduce participation barriers by making training fees more affordable and accessible, effectively improve the comprehensive quality and training satisfaction of laboratory animal practitioners, achieve better teaching outcomes, and provide replicable and promotable practical references for the standardized upgrading of the training system in the laboratory animal industry and other industries.

Objective:To investigate the risk factors associated with post-prematurity respiratory disease (PPRD) in very preterm infants.Methods:A prospective cohort study was conducted, enrolling 369 very preterm infants who were admitted to the neonatal intensive care unit of Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, within one week of birth from January 2019 to June 2023. Data on maternal and infant clinical characteristics, neonatal morbidities, and treatments during hospitalization were collected. The very preterm infants were divided into 2 groups based on whether they developed PPRD. Continuous variables were compared using Mann-Whitney U test, while categorical variables were compared using χ2 tests or continuity correction χ2 test. Multivariate Logistic regression analysis was used to identify the independent risk factors for PPRD in very preterm infants. Results:Among the 369 very preterm infants, 217 cases(58.8%) were male, with a gestational age of 30 (28, 31) weeks at birth and a birth weight of 1 320 (1 085, 1 590) g. Of these, 116 cases (31.4%) developed PPRD, while 253 cases (68.6%) did not. The very preterm infants in the PPRD group had a lower gestational age and lower birth weight (both, P<0.001). The PPRD group also had a higher proportion of males, lower Apgar scores at the 1 th minute after birth and the 5 th minutes after birth, a higher rate of born via cesarean delivery, and a higher incidence of bronchopulmonary dysplasia, more pulmonary surfactant treatment, longer durations of mechanical ventilation, longer total oxygen therapy, and lower Z-score for weight at discharge (all P<0.05). Multivariate Logistic regression analysis showed that gestational age ( OR=0.85, 95% CI 0.73-0.99, P=0.037), born via cesarean delivery ( OR=2.23, 95% CI 1.21-4.10, P=0.010), a duration of mechanical ventilation ≥7 days ( OR=2.51, 95% CI 1.43-4.39, P=0.001), and a Z-score for weight at discharge ( OR=0.82, 95% CI 0.67-0.99, P=0.040) were all independent risk factors for PPRD in very preterm infants. Conclusion:Very preterm infants with a small gestational age, born via cesarean section, mechanical ventilation ≥7 days, and a low Z-score for weight at discharge should be closely monitored for PPRD, and provided with standardized respiratory management after discharge.

Objective To investigate the effects and mechanisms of high-fat diet(HFD)-induced obesity on male ze-brafish reproductive function.Methods Adult male zebrafish were divided into normal diet(ND)group and HFD group.Growth and metabolic conditions were evaluated by measuring body weight,body length,BMI,organ index,and glucose/lipid lev-els.Reproductive capacity was assessed via sperm concentration,motility,fertilization rate,and testosterone levels.Testicular tissues from both of groups were subjected to transeriptomic sequencing(RNA-seq).And qRT-PCR was used to validate the expression of genes.Results Compared to male zebrafish in ND group,the ones in HFD group exhibited hepatic steatosis and glucose/lipid meta-bolic disorders(P＜0.05).Testicular structural disorganization,along with reduced testosterone levels,decreased gonadosomatic in-dex,and impaired sperm concentration and motility occurred in HFD group(P＜0.05).GO analysis revealed that differentially ex-pressed genes were enriched in spermatogenesis and ciliary system,while KEGG analysis highlighted metabolic related pathways(pu-rine metabolism,thyroid hormone synthesis,mTOR signaling)and cell adhesion molecules.Twenty key differentially expressed genes were validated by qRT-PCR,which confirmed the reliability of RNA-Seq results.Conclusion Impairment of reproductive function induced by HFD in zebrafish may be associated with three regulatory mechanisms including ciliary system,metabolic dysregulation,and aberrant cell adhesion molecule signaling.This study provides mechanistic insights and identifies potential therapeutic targets for clinical management of diet-associated infertility.

Objective To explore the efficacy of precise surface positioning method in percutaneous screw fixation for Jones fracture.Methods Data of 46 patients with Jones fractures admitted to our hospital from September 2021 to December 2023 were retrospectively analyzed.According to the different treatment methods,the patients were divided into screw groups(24 patients fixed by percutaneous screws)and tension band groups(22 patients fixed by incision and reset tension band).The general conditions,follow-up time,surgical time,hospitalization time,fracture reduction quality,postoperative complete weight-bearing time,fracture healing time,and complications in the two groups were recorded and compared.The American Association of Foot and Ankle Surgery(AOFAS)system was used to assess the recovery of foot function.Results There was no statistically significant difference in the general data before the operation between the two groups of patients(P＞0.05).The follow-up time of the two groups was 6 to 24 months.Compared with the tension band group,the screw group had shorter operation time[(32.96±5.35)min vs.(47.27±9.42)min],shorter hospitalization time[(5.75±1.18)d vs.(7.68±2.05)d],earlier postoperative complete weight-bearing time[(6.47±0.73)w vs.(7.29±1.66)w],faster fracture healing time[(3.31±0.68)months vs.(4.29±1.29)months],higher AOFAS score[(89.13±2.62)vs.(79.59±4.49)](all P＜0.05).The tension band group of fracture reduction quality was better than the screw group(P＜0.05).The quality of fracture reduction during the operation in the tension band group was better than that in the screw group(P＜0.05).In terms of complications:no complications occurred in the screw group,one patient in the tension band group developed infection,and one patient had a loose Kirchner needle.Conclusion Precise surface positioning method in percutaneous screw fixation for Jones fracture is a safe and effective method with precise positioning,simple operation,small trauma,and rapid rehabilitation,which is worthy of clinical promotion,but the indications need to be strictly controlled according to the type of fracture.

BACKGROUND:In vitro reconstruction of hair follicles depends on the self-assembly behavior of dermal papilla cells and keratinocytes during three dimensional co-culture,which is very similar to the development of hair follicle embryos.Therefore,hair follicle organoids can be used as a good model in vitro for basic research of hair follicle regeneration.Currently,wholly humanized follicle organoids cultured by biomimetic system have been successfully transplanted into animals.Research on hair follicle organoids makes it possible to regenerate hair follicles,but there is no uniform conclusion on the construction method,and there are still many challenges.OBJECTIVE:To review the current research progress of hair follicle organoids,summarize their culture methods and characterization,and expound their application prospects,so as to provide reliable basis for subsequent research,in order to promote its clinical transformation in tissue engineering and alopecia treatment.METHODS:CNKI and PubMed databases were searched by a computer,using"hair follicle regeneration,hair regeneration,organoid"as Chinese search terms and"hair growth,hair loss,hair regeneration,hair follicle,dermal papilla cell,organoid"as English search terms.The domestic and foreign literature on hair follicle organoids was reviewed and a total of 67 articles were included for follow-up analysis.RESULTS AND CONCLUSION:(1)Dermal papilla cells and keratinocytes from different sources can be used to construct hair follicle organoids in vitro to obtain mouse-derived,human-mouse chimeric and human-derived hair follicle organoids,which have been successfully used in rodent transplantation models.(2)A variety of programming methods,such as electrical stimulation,addition of recombinant protein,and dissociation of cell gene expression,can be used to promote hair follicle organoid such ashair follicle germ formation and hair shaft elongation.Simultaneously,the application of various 3D printing technologies has achieved controllable hair follicle organoid to the size of the quantity production,which greatly improves the production efficiency.(3)When melanocytes are added to the dissociated cell co-culture system,the problem of pigment deficiency in new hair follicles may be solved.When human umbilical vein endothelial cells are added,the new blood vessels in the cell aggregates can improve their nutrient supply and reduce necrosis in the central area.(4)As the technology of engineered hair follicle organoid gradually maturates,hair follicle organoids constructed from patient's autologous cells may be used for individualized treatment of severe alopecia.

BACKGROUND:The blood-brain barrier is an important structure that protects the central nervous system,and the study of the effects of high glucose on the blood-brain barrier is important for the prevention of high glucose-induced damage to the central nervous system.OBJECTIVE:To investigate the potential effect of high glucose on the blood-brain barrier function of hCMEC/D3 human brain microvascular endothelial cells.METHODS:hCMEC/D3 cells were cultured in regular sugar medium(glucose concentration of 25 mmol/L)and high-sugar medium(glucose concentration of 55 mmol/L).The morphology of cells in each group was observed by light microscopy.CCK-8 assay was used to detect changes in cell viability.A monolayer blood-brain barrier model was established using hCMEC/D3 cell line with Transwell chamber device.Changes in cell transmembrane resistance were monitored daily.The permeability of cell monolayers was detected by phenol red permeability.Flow cytometry was used to detect the apoptosis rate of the cells.Western blot assay was used to detect the expression of Bcl-2,Bax,Caspase-3,ZO-1,Occludin,Claudin-1,and histone deacetylase 4.The levels of histone deacetylase in cell supernatant were detected by ELISA.The expression of histone deacetylase 4 in cells was detected by immunofluorescence.RESULTS AND CONCLUSION:(1)The cell viability of high sugar group was significantly lower than that of control group(P＜0.000 1).(2)The cells of the control group were in a good growth state,interwoven into a dense mesh,with interconnections between synapses.The cell growth of high glucose group was suppressed,and the connection of inter-cellular synapses was reduced.(3)Compared with the control group,the transmembrane resistance value of the high glucose group was reduced(P＜0.05);phenol-red permeability of the monolayer cell membrane was increased(P＜0.05);cell apoptosis rate was increased(P＜0.01);the expression of Bax protein was increased(P＜0.000 1);the expression of Caspase-3 protein had no significant change(P＞0.05);the expression of Bcl-2,ZO-1,Occludin,Claudin-1,and histone deacetylase 4 proteins was decreased(P＜0.01,P＜0.001,P＜0.01,P＜0.000 1,P＜0.01);the fluorescence expression of histone deacetylase 4 was decreased(P＜0.001)in the high glucose group.(4)The level of histone deacetylase 4 in the cell supernatant of the high glucose group was lower than that of the control group(P＜0.05).The results show that high glucose induces the increased apoptosis and enhances permeability of hCEMCE/D3 cells,and its mechanism may be related to the decreased expression level of histone deacetylase 4.