Spermatogenesis is a highly ordered and spatiotemporally regulated developmental process in the male reproductive system, during which spermatogonial stem cells (SSCs), supported by the seminiferous tubule microenvironment, sequentially undergo mitosis, meiosis, and spermiogenesis to ultimately generate structurally intact spermatozoa. This complex process is accompanied by extensive transcriptional reprogramming, chromatin remodeling, and finely tuned post-transcriptional regulation. Precise control of RNA fate is therefore essential for maintaining the continuity and fidelity of spermatogenesis, and its disruption represents a major molecular basis of male infertility. N⁶-methyladenosine (m⁶A), the most abundant internal RNA modification in eukaryotes, has emerged as a critical regulator of post-transcriptional gene expression. m⁶A methyltransferases (“writers”) catalyze the addition of a methyl group to the N⁶ position of adenosine, m⁶A demethylases (“erasers”) remove the modification, and m⁶A-binding proteins (“readers”) recognize m⁶A-modified transcripts. Through the coordinated actions of these factors, m⁶A regulates transcript fate at multiple levels, including RNA splicing, nuclear export, stability, translation, and decay. Emerging evidence indicates that m⁶A-mediated regulation is essential across multiple stages of spermatogenesis, including SSC self-renewal and differentiation, meiotic progression, maintenance of chromosomal stability, and sperm morphogenesis. Beyond its intrinsic functions in germ cells, m⁶A also contributes to the regulation of the testicular microenvironment. In sertoli cells, m⁶A is involved in maintaining blood-testis barrier integrity, RNA processing, and paracrine signaling, thereby providing structural and metabolic support for germ cell development. In Leydig cells, m⁶A regulates steroidogenesis, particularly testosterone synthesis, and participates in cellular stress responses and metabolic homeostasis. Through these mechanisms, m⁶A indirectly influences spermatogenesis by modulating the functional state of testicular somatic cells, highlighting an integrated regulatory mode that combines cell-intrinsic and microenvironment-mediated effects. Notably, distinct classes of m⁶A regulators exhibit pronounced stage-specific functions and coordinated division of labor, collectively forming a multilayered and dynamic regulatory network. Writers often display dosage- and temporal window-dependent effects; erasers contribute to stage-specific demethylation and functional compensation; while readers function through a “switch-buffer” dual-layer architecture, and RNA-binding proteins (RBPs) participate in substrate selection and post-transcriptional regulation. Importantly, emerging evidence suggests that some m⁶A-related proteins can function through noncanonical mechanisms independent of m⁶A recognition, such as intrinsic RNA-binding activity, helicase function, or ribonucleoprotein complex assembly, thereby expanding the functional landscape of the m⁶A regulatory system. Dysregulation of m⁶A machinery can lead to multiple spermatogenic defects, including impaired SSC self-renewal, meiotic arrest, abnormal chromatin remodeling, and defective sperm formation, ultimately resulting in male infertility. Despite substantial advances, several critical questions remain unresolved, including the distinction between m⁶A-dependent and -independent mechanisms, the spatiotemporal dynamics of m⁶A modifications at single-cell resolution, and the coordination and antagonism among different regulatory factors. In this review, we systematically summarize the dual regulation of spermatogenesis by germ cell-intrinsic mechanisms and the testicular microenvironment, and delineate the molecular mechanisms and stage-specific functions of the dynamic m⁶A regulatory network. We further discuss the current limitations in the field and propose feasible experimental strategies for future investigation. Collectively, this work aims to provide a comprehensive framework for understanding the epitranscriptomic regulation of spermatogenesis and to offer theoretical insights into the pathogenesis and clinical management of male infertility.

Spermatogenesis is a highly ordered and spatiotemporally regulated developmental process in the male reproductive system, during which spermatogonial stem cells (SSCs), supported by the seminiferous tubule microenvironment, sequentially undergo mitosis, meiosis, and spermiogenesis to ultimately generate structurally intact spermatozoa. This complex process is accompanied by extensive transcriptional reprogramming, chromatin remodeling, and finely tuned post-transcriptional regulation. Precise control of RNA fate is therefore essential for maintaining the continuity and fidelity of spermatogenesis, and its disruption represents a major molecular basis of male infertility. N⁶-methyladenosine (m⁶A), the most abundant internal RNA modification in eukaryotes, has emerged as a critical regulator of post-transcriptional gene expression. m⁶A methyltransferases (“writers”) catalyze the addition of a methyl group to the N⁶ position of adenosine, m⁶A demethylases (“erasers”) remove the modification, and m⁶A-binding proteins (“readers”) recognize m⁶A-modified transcripts. Through the coordinated actions of these factors, m⁶A regulates transcript fate at multiple levels, including RNA splicing, nuclear export, stability, translation, and decay. Emerging evidence indicates that m⁶A-mediated regulation is essential across multiple stages of spermatogenesis, including SSC self-renewal and differentiation, meiotic progression, maintenance of chromosomal stability, and sperm morphogenesis. Beyond its intrinsic functions in germ cells, m⁶A also contributes to the regulation of the testicular microenvironment. In sertoli cells, m⁶A is involved in maintaining blood-testis barrier integrity, RNA processing, and paracrine signaling, thereby providing structural and metabolic support for germ cell development. In Leydig cells, m⁶A regulates steroidogenesis, particularly testosterone synthesis, and participates in cellular stress responses and metabolic homeostasis. Through these mechanisms, m⁶A indirectly influences spermatogenesis by modulating the functional state of testicular somatic cells, highlighting an integrated regulatory mode that combines cell-intrinsic and microenvironment-mediated effects. Notably, distinct classes of m⁶A regulators exhibit pronounced stage-specific functions and coordinated division of labor, collectively forming a multilayered and dynamic regulatory network. Writers often display dosage- and temporal window-dependent effects; erasers contribute to stage-specific demethylation and functional compensation; while readers function through a “switch-buffer” dual-layer architecture, and RNA-binding proteins (RBPs) participate in substrate selection and post-transcriptional regulation. Importantly, emerging evidence suggests that some m⁶A-related proteins can function through noncanonical mechanisms independent of m⁶A recognition, such as intrinsic RNA-binding activity, helicase function, or ribonucleoprotein complex assembly, thereby expanding the functional landscape of the m⁶A regulatory system. Dysregulation of m⁶A machinery can lead to multiple spermatogenic defects, including impaired SSC self-renewal, meiotic arrest, abnormal chromatin remodeling, and defective sperm formation, ultimately resulting in male infertility. Despite substantial advances, several critical questions remain unresolved, including the distinction between m⁶A-dependent and -independent mechanisms, the spatiotemporal dynamics of m⁶A modifications at single-cell resolution, and the coordination and antagonism among different regulatory factors. In this review, we systematically summarize the dual regulation of spermatogenesis by germ cell-intrinsic mechanisms and the testicular microenvironment, and delineate the molecular mechanisms and stage-specific functions of the dynamic m⁶A regulatory network. We further discuss the current limitations in the field and propose feasible experimental strategies for future investigation. Collectively, this work aims to provide a comprehensive framework for understanding the epitranscriptomic regulation of spermatogenesis and to offer theoretical insights into the pathogenesis and clinical management of male infertility.

ObjectiveTo investigate the effects of metformin on the immune functions of immature dendritic cells （imDCs） and the underlying mechanisms. MethodsMouse bone marrow-derived imDCs were treated with different concentrations of metformin. The working concentration and treatment time of metformin in this study were determined based on the results of cell apoptosis and cell viability assays. The effects of metformin on the phagocytic capacity of imDCs was evaluated using an antigen endocytosis assay. The expression of cluster of differentiation 205 （CD205）， the polymerization of filamentous actin （F-actin）， and the underlying regulatory mechanisms were investigated through flow cytometry， laser confocal fluorescence microscopy， and Western blot. ResultsThe working concentrations of metformin were 1， 2， 4 mmol/L for 24 h determined by the apoptosis and cell viability assays.Metformin significantly suppressed the phagocytic capacity of imDCs， down-regulated the expression of the mannose receptor CD205 on the cell surface， which was closely associated with phagocytic function； metformin inhibited the RhoA-ROCK1-LIMK1-Cofilin signaling pathway， which inhibited the polymerization of F-actin and disturbed its dynamic remodeling of imDCs. ConclusionMetformin can inhibit the expression of CD205 and disrupt the remodeling of F-actin， thereby suppressing the antigen-capturing capacity of imDCs.

BackgroundDepression in mid-to-late pregnancy has significant negative effects on pregnant women and their offspring. Previous studies have shown that pregnant women with depression in mid-to-late pregnancy exhibit lower frontal lobe activation levels， but the activation phase and its impact on depression require further investigation. ObjectiveTo analyze the functional near-infrared spectroscopy （fNIRS） features of depressed pregnant women in mid-to-late pregnancy， and to provide references for assisting in the clinical screening of depression in them. MethodsA total of 40 pregnant women in mid to late pregnancy with Edinburgh Postnatal Depression Scale （EPDS） score above 11 who underwent prenatal examination at the department of obstetrics of the Third Hospital of Mianyang from September 2023 to July 2024 were included as the study group. During the same period， 40 pregnant women in mid-to-late pregnancy with EPDS score below 11 who were matched with the age， family history， and past history of the study group were recruited as the control group. A self-designed questionnaire was used to collect basic information of the pregnant women. The fNIRS technique was used to measure the prefrontal and temporal lobe integral values and center of gravity values during the verbal fluency task （VFT） pregnant women in mid-to-late pregnancy. Spearman correlation analysis was conducted to examine the correlation between EPDS scores and fNIRS imaging features， as well as demographic characteristics， in mid-to-late pregnant women with depression. Multiple linear regression was used to analyze the influencing factors of depression symptoms in this population. ResultsStatistically significant differences were observed between the study group and the control group in terms of educational level and body mass index （BMI） （χ²/t=4.528， 2.292， P<0.05）. The study group demonstrated a lower prefrontal integral value and a higher prefrontal center of gravity value compared with the control group （t=-7.601， 5.641， P<0.05）. EPDS scores of depressed pregnant women in mid-to-late pregnancy were negatively correlated with prefrontal lobe integral value （r=-0.680， P<0.01）， while positively correlated with prefrontal lobe center of gravity value and BMI （r=0.737， 0.604， P<0.01）. Multiple linear regression analysis showed that age （β=-0.214， P<0.01）， BMI （β=0.340， P<0.01）， prefrontal integral value （β=-0.351， P<0.01）， and prefrontal center of gravity value （β=0.347， P<0.01） were influencing factors of depressive symptoms in pregnant women in mid-to-late pregnancy. ConclusionThe prefrontal lobe activation degree of depressed pregnant women in mid-to-late pregnancy is lower than that of non-depressed pregnant women， and the activation phase is delayed. Age， prefrontal lobe integral value， prefrontal center of gravity value and BMI may be the influencing factors of depressive symptoms in pregnant women in the middle and late pregnancy.

Objective:To investigate the impact of different etiologies on the spatial distribution pattern of infarcts and the mapping pattern of focal symptoms in acute ischemic stroke (AIS) using a population-based standardized spatial analysis of MRI.Methods:This was a cross-sectional study. Clinical [age, sex distribution, admission National Institutes of Health Stroke Scale (NIHSS) score and 90-day modified Rankin Scale (mRS) score at discharge, etc.] and imaging data of 2 610 patients with AIS attending 9 Medical Centers from January 2015 to December 2021 were retrospectively analyzed. All patients were categorized into 1 718 cases of large artery atherosclerosis (LAA) type, 335 cases of cardioembolism (CE) type, and 557 cases of small artery occlusion (SAO) type according to TOAST typing. All patients underwent diffusion-weighted imaging, and the detected infarct lesions were segmented and aligned to the standardized space using artificial intelligence-assisted methods, and the spatial distribution frequency heatmaps of lesion locations in patients with different TOAST subtypes were plotted and compared with each other by χ2 test. Lesion-symptom image brain maps with different clinical symptoms were further plotted, and differences of lesion-symptom image relationships among different TOAST subtypes were observed and compared with each other by interaction effect. Results:In all patients, the favored sites of infarct lesions were the bilateral middle cerebral artery region in the anterior circulation and the occipital and brainstem regions in the posterior circulation. Compared with the LAA type, the CE type lesions were more likely to occur in the anterior cerebral artery region, the occipital lobe, and the cerebellum posterior, while the SAO type lesions were more likely to occur in the perforator artery supply area. The lesion-symptom mapping results showed that AIS patients with infarct lesions in the frontoparieto-temporal region in the presence of a left middle cerebral artery supply had higher admission NIHSS scores and higher discharge 90-day mRS scores for the LAA type than for the CE type( P<0.05); AIS patients with infarcted lesions in the brainstem region and some cerebellar regions in the presence of vertebrobasilar artery supply had higher admission NIHSS scores and higher discharge 90-day mRS scores for the CE type than for the LAA type( P<0.05). Conclusion:At the population level, brain mapping reveals specific infarct distribution patterns and differences in lesion-symptom mapping patterns of different etiologies AIS patients, providing imaging evidence for the understanding of AIS pathogenetic mechanisms and clinical management.

With the rapid development of generative artificial intelligence(GAI)technologies,their widespread application in academic research and writing is continuously expanding the boundaries of sci-entific inquiry.However,this trend has also raised a series of ethical and regulatory challenges,inclu-ding issues related to authorship,content authenticity,citation accuracy,and accountability.In light of the growing involvement of AI in generating academic content,establishing an open,controllable,and trustworthy ethical governance framework has become a key task for safeguarding research integrity and maintaining trust within the academic community.This expert consensus outlines ethical requirements across key stages of AI-assisted academic writing-including topic selection,data management,citation practices,and authorship attribution.It aims to clarify the boundaries and ethical obligations surrounding AI use in academic writing,ensuring that technological tools enhance efficiency without compromising in-tegrity.The goal is to provide guidance and institutional support for building a responsible and sustainable research ecosystem.

Objective:To investigate the differences in prosthesis implantation accuracy, biomechanical indicators, and 2-year postoperative clinical outcomes between robotic-assisted traditional posterior approach (RA-PLA) and robotic-assisted direct anterior approach (RA-DAA) in total hip arthroplasty (THA).Methods:This study is a prospective randomized controlled trial. Patients with unilateral femoral head ischemic necrosis or primary hip osteoarthritis who meet the inclusion and exclusion criteria and were admitted to the Department of Orthopedics,Xinqiao Hospital, Army Medical University from May to September 2022. Divided into RA-PLA group and RA-DAA group through central randomization. Use cumulative and fitting methods to analyze the learning curve of robotic surgery and eliminate cases before the inflection point of the learning curve. Compare the abduction angle and anteversion angle of acetabular cup implantation between two groups of patients, as well as the rate of falling into the safe zone, the difference in length between the two lower limbs, hip joint eccentricity, rotation center height, the complete accuracy of prosthesis planning (the number of cases in which acetabular cup prosthesis, femoral stem prosthesis specifications, and neck shaft angle were completely consistent with surgical planning during surgery/total cases×100%), patient reported outcome indicators (including Harris hip score (HHS), Western Ontario and McMaster Universities Osteoarthritis Index(WOMAC), and 12 item Short Form Health Survey (SF-12) score) and clinical outcomes. Data comparison was conducted using independent sample t-test, Wilcoxon rank sum test, chi square test, Fisher′s exact probability method, or repeated measures analysis of variance. Results:A total of 98 patients were included in this study, with 48 in the RA-PLA group and 50 in the RA-DAA group. After excluding cases before the inflection point of the learning curve, 30 patients were included in each group. There was no statistically significant difference in baseline data between the two groups before surgery (all P>0.05). There was no significant difference in the values of the anterior tilt angle and abduction angle between the two groups of acetabular cups compared to preoperative planning (all P>0.05). The proportion of patients who fell into the Lewinnek and Callanan safe zones was 90.0% (27/30) and 96.7% (29/30), respectively ( χ2=0.268, P=0.605). There was no significant difference in postoperative lower limb length, hip joint eccentricity, and rotation center height deviation and grading between the two groups (all P>0.05). The complete accuracy of prosthesis planning in the RA-DAA group was higher than that in the RA-PLA group (86.7% (26/30) compared to 63.3% (19/30), χ2=4.356, P=0.037).All patients were followed up for more than 2 years. In terms of postoperative HHS, WOMAC, and SF-12 score, there was no statistically significant difference in the inter group effect comparison (all P>0.05), but there was a significant statistical significance in the time effect (all P<0.05). There was no significant difference in the incidence of perioperative complications and adverse events between the RA-DAA group and the RA-PLA group (20.0% (6/30) vs. 13.3% (4/30), χ2=0.480, P=0.488). Conclusions:RA-DAA and RA-PLA techniques can achieve similar clinical efficacy after two years of surgery, and both can achieve similar reconstruction accuracy in terms of acetabular cup implantation angle, lower limb length, hip joint eccentricity, and rotation center height. The accuracy of prosthesis planning in RA-DAA is higher.

Abnormal signaling in the androgen receptor(AR)signaling pathway is critical for prostate cancer development and progression,so inhibition of AR activity through androgen deprivation therapy(ADT)is an important means to control the development of prostate cancer in the early stage.However,most patients relapse and develop castrate-resistant prostate cancer(CRPC)within 6～20 months.Sur-gery and radiotherapy are still the major treatments for CRPC,but there are adverse effects such as urina-ry symptoms and sexual dysfunction.The first and second generatiosn of novel AR inhibitors can effec-tively treat CRPC.However,resistance to these chemicals is inevitable,and thus many patients may ex-perience recurrence.Resistance to AR inhibitors mainly consists of AR mutations,splice variant forma-tion and amplification,which have been shown to play an important role in CRPC.Also,aberrant activa-tion of cyclin dependent kinase(CDKs)and epigenetic alterations(e.g.histone modifications and DNA methylation)have been reported to be associated with prostate cancer progression.Proteolysis targeting chimeras(PROTACs)have unique advantages in CRPC therapy by virtue of their unique mechanism of action,ability to target non-druggable proteins,and specific binding to targets.In this review,we sum-marize the development of PROTAC technology for the treatment of CRPC by targeting different structural domains of AR,CDKs and epigenetic markers,and discuss the future prospects and challenges of PRO-TACs in the therapeutic field.

Objective:To observe the clinical efficacy and effects on emotional state, anorectal physiological function, serum inflammation factors and neurotransmitters of repetitive transcranial magnetic stimulation (rTMS) on functional anorectal pain (FAP) patients, and to explore the potential therapeutic mechanisms.Methods:From September 1, 2022 to December 31, 2023, a total of 50 FAP patients who were admitted to Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine were enrolled in this study. The patients were randomly divided into the treatment group (20 cases) and the control group (20 cases) according to a random number table and relevant exclusion criteria. The treatment group received rTMS treatment and the control group received sham rTMS treatment. The Hamilton anxiety scale (HAMA) score, Hamilton depression scale (HAMD) score, visual analogue scale (VAS) score, high-resolution anorectal manometry data (anal resting pressure, anal squeeze pressure, initial sensation shreshold, defecation shreshold, defecation urgency shreshold, and tolerance shreshold), and the levels of serum inflammatory factors (interleukin(IL)-4, IL-8, tumor necrosis factor-α) and 5-hydroxytryptamin(5-HT) were recorded before and after treatment. Independent sample t-test, paired t-test, Mann-Whitney U test and Wilcoxon signed-rank test were used for statistical analysis. Results:The VAS, HAMA, and HAMD scores of the treatment group after treatment were lower than those before treatment (3.85±2.23 vs. 6.85±1.98, 4.40±3.39 vs. 8.75±6.60, and 7.10±6.56 vs. 12.85±7.20), and were also lower than those of the control group after treatment(6.50±1.76, 8.20±6.65, 12.10±6.80), and the differences were statistically significant ( t=5.68, 4.72, 6.06; -4.17, -2.27, -2.37; P<0.001, <0.001, <0.001; <0.001, =0.028, and =0.023). The initial sensation shreshold, defecation shreshold, defecation urgency shreshold, and tolerance shreshold of the treatment group after treatment were higher than those before treatment(30.00(30.00, 46.00) mmHg (1 mmHg=0.133 kPa) vs. 23.00(18.50, 29.00) mmHg, 50.00(44.50, 60.00) mmHg vs. 37.00(30.75, 51.50) mmHg, (74.30±16.02) mmHg vs. (63.70±22.21) mmHg, 119.00(100.00, 148.00) mmHg vs. 98.00 (69.50, 153.00) mmHg), and the tolerance shreshold of the treatment group after treatment was higher than that of the control group after treatment(119.00 (100.00, 148.00) mmHg vs. 102.00(84.50, 111.50) mmHg), and the differences were statistically significant ( Z=–3.14 and –2.86, t=-4.02, Z=-2.84 and -2.11; P=0.002, 0.004, 0.001, 0.004, and 0.035). Additionally, the 5-HT level of the treatment group after treatment was higher than that before treatment (1 549.41 (1 320.21, 1 640.03) μg/L vs. 1 081.52(874.36, 1 626.79) μg/L), and the difference was statistically significant ( Z=-2.88, P=0.004). Conclusion:The rTMS treatment can effectively relieve the pain, anxiety and depression, improve visceral sensitivity, and influence the neurotransmitter level of brain-gut axis in FAP patients.

This study was aimed at preliminarily investigating the genetic and antimicrobial resistance characteristics of Salmonella Alachua isolates through whole-genome analyses.Five Salmonella Alachua isolates from various sources(both hu-man and non-human)were collected and identified.Phenotype and serotype verification,antimicrobial susceptibility testing,and whole-genome sequencing were performed.Virulence genes,antimicrobial resistance genes,and plasmid replicons were predicted according to globally available Salmonella Alachua genomic data.A phylogenetic tree was constructed to explore the genetic background.The first report of Salmonella Alachua in China emerged in Shanghai in 2015,and patients presented pri-marily with diarrhea.The isolates have been found predominantly in the eastern and southern coastal regions.Among the five i-solates analyzed,four belonged to sequence type(ST)2061,and one belonged to ST1298.All isolates were susceptible to most commonly used clinical antibiotics.Whole-genome analyses revealed that two ST2061 strains carried the blaKPC-2 gene,and one ST1298 strain carried the fosA7 gene.Phylogenetic analysis of global Salmonella Alachua populations indicated that the ST2061 clone belonged to the C1 clade,which was closely related to strains from the UK,whereas the ST1298 clone was found in the C4 clade,a globally disseminated fosA 7-positive lineage.This study provides initial insights into the genetic and antimi-crobial resistance characteristics of Salmonella Alachua in China,highlighting the presence of strains carrying blaKPC-2 and fo-sA7 genes.These findings may provide a reference for future large-scale molecular epidemiological surveillance and source-trac-ing efforts,and they underscore the importance of enhanced resistance monitoring for Salmonella Alachua.