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.

To investigate the molecular mechanisms underlying the mechanosensitive ion channel PI-EZO2 in osteoarthritis(OA),we developed a lentiviral vector for endogenous PIEZO2 overexpression and established a stable PIEZO2-high-expressing immortalized human primary chondrocyte line.By map-ping the open reading frame of the PIEZO2 locus and designing sequence-specific sgRNA,we employed the CRISPR/Cas9 synergistic activation mediator(SAM)system to precisely integrate transcriptional ac-tivation elements into the PIEZO2 promoter region.Lentiviral-mediated targeted genomic integration en-sured endogenous PIEZO2 overexpression,confirmed by mCherry fluorescence tracing coupled with flow cytometric sorting,which revealed membrane-specific localization of PIEZO2 protein(localization effi-ciency:78.49％).Quantitative PCR demonstrated a 17-fold upregulation of PIEZO2 mRNA,while Western blotting validated enhanced membrane-localized protein expression.Strikingly,PIEZO2-overex-pressing chondrocytes exhibited hallmark OA metabolic phenotypes compared to wild-type controls:typeⅡ collagen mRNA expression decreased to 50％of baseline levels,whereas matrix metalloproteinase 13(MMP13)mRNA surged by 20-fold.These alterations recapitulated the pathological matrix metabolic phenotype observed in biomechanical OA models induced by cyclic mechanical stress(10％strain,0.5 Hz,8 h/day for 2 consecutive days).Collectively,we successfully generated a human chondrocyte model with stable PIEZO2 overexpression,which faithfully mirrors mechanotransduction-driven OA progression.This engineered cellular system provides a robust platform for dissecting PIEZO2-mediated mechanosig-naling networks and advancing targeted therapeutic discovery.

Head and neck squamous cell carcinoma(HNSCC)is the most common pathological subtype of head and neck malignancies.The majority of patients are diagnosed at an advanced stage,losing the opportunity for curative surgery,and overall prognosis remains poor.With its exceptional capabilities in image analysis and powerful data pro-cessing,artificial intelligence(AI)has the potential to become a valuable tool in precision pathology,offering new op-portunities to improve the diagnosis and treatment of head and neck cancer.This article provides a systematic review of the current research status on AI-assisted pathological image analysis in the diagnosis,treatment strategy selection,and prognosis prediction of HNSCC,while also discussing future research directions.

Objective:To evaluate the effect of radiotherapy on hypothyroidism (HT) in patients with breast cancer after radical surgery, and to establish a HT prediction model.Methods:Clinical data of 296 patients who received radiotherapy after radical mastectomy and followed up regularly in Affiliated Cancer Hospital of Xinjiang Medical University from January 2015 to May 2023 were retrospectively analyzed. Baseline clinicopathological information was extracted and thyroid dose volume histograms were reviewed to obtain dosimetric parameters such as thyroid volume, thyroid D mean, D min, D max and V 15 Gy-V 50 Gy, etc. The thyroid function levels of all patients were detected before radiotherapy and at 3, 6, 9, 12, 18, 24, 36, 48 and 60 months after radiotherapy, and the incidence of HT after treatment was calculated to determine the effect of radiotherapy on thyroid function. Cox regression model was used for univariate and multivariate analyses to identify the factors associated with HT and establish the predictive model. The optimal cut-off value of influencing factors was calculated by using the receiver operating characteristic curve. The cumulative incidence curve was drawn by Kaplan-Meier method. Inter-group differences were compared by using log-rank test. Results:At a median follow-up of 36 months, HT occurred in 130 patients (43.92%). The peak of HT occurred approximately 12 (9-18) months after radiotherapy. Univariate analysis showed that neoadjuvant chemotherapy ( HR=1.736, 95% CI=1.222-2.467, P=0.002), thyroid D min ( HR=1.001, 95% CI=1.001-1.001, P<0.001) and thyroid D mean ( HR =1.001, 95% CI=1.001-1.002, P<0.001) were the risk factors for HT. Multivariate analysis showed that thyroid D mean was correlated with the occurrence of HT ( HR=1.001, 95% CI=1.001-1.001, P=0.002). The optimal cut-off value of thyroid D mean was 2771.66 cGy, and the incidence of HT was statistically different between high-risk group (D mean>2771.66 cGy) and low-risk group (D mean≤2771.66 cGy) ( P<0.001). Conclusions:Radiotherapy after radical surgery is associated with the occurrence of HT in patients with breast cancer. Minimizing the thyroid D mean below 2771.66 cGy is an effective method to protect thyroid function. It is recommended to start monitoring thyroid function no later than 9 months after radiotherapy in patients receiving neoadjuvant chemotherapy.

Objective:To investigate the impact of endocrine therapy(ET)on the prognosis of breast cancer patients with low estrogen re-ceptor(ER)expression and to identify the associated independent prognostic factors.Methods:We retrospectively analyzed the clinicopath-ological data of 387 female patients with breast cancer with low ER expression and at stage Ⅰ-Ⅲ TNM who were treated surgically at Xinji-ang Medical University Affiliated Tumor Hospital between January 2014 and May 2020.The patients were stratified into ET and non-ET groups based on whether they received ET administration.Propensity score matching(PSM)was used to balance intergroup differences,and survival analysis along with Cox proportional hazards models were applied to assess prognosis and the associated independent influencing factors.Results:Among the 387 patients,214(55.3%)received ET and 173(44.7%)did not.Baseline data indicated that the non-ET group exhibited higher proportions of ethnic minorities and patients with regional lymph node involvement,advanced TNM stage,and no radio-therapy administration.After PSM,baseline characteristics showed no significant differences between the ET(n=132)and non-ET groups(n=132).Pre-PSM survival analysis revealed 5-year overall survival(OS)rates of 93.4%and 72.8%and 5-year disease-free survival(DFS)rates of 74.1%and 60.9%for the ET and non-ET groups,respectively;both rates showed statistically significant differences between the groups(P<0.05).Post-PSM,the 5-year OS rates significantly differed between the ET and non-ET groups(91.6%and 80.1%,respectively;P<0.05),whereas no significant difference in 5-year DFS rates were observed(71.1%[ET group]vs.68.1%[non-ET group],P>0.05).Multivariate ana-lysis identified the following independent prognostic factors:for OS,ET administration,CK5/6 status,tumor size T3,lymph node status N2/N3,and radiotherapy administration(all P<0.05).For DFS,the independent factors included CK5/6 status,tumor size T3,lymph node status N2/N3,radiotherapy administration,and targeted therapy(all P<0.05).Conclusions:Endocrine therapy significantly improved the 5-year OS in breast cancer patients with low ER expression.CK5/6 positivity,large tumor size(T3),and regional lymph node involvement(N2/N3)were independent risk factors for both 5-year OS and DFS.Adjuvant radiotherapy and trastuzumab-based therapy significantly im-proved prognosis.

Purpose To investigate the consistency and reliability of immunohistochemistry(IHC)staining and polymerase chain reaction-capillary electrophoresis(PCR-CE)in the detection of microsatellite instability.Methods The clinical data of 735 patients with solid tumors who underwent biopsy or surgical resection were collected,and the microsatellite status of tissue samples was detected by IHC and PCR-CE.Results Among the 344 cases of colorectal cancer(CRC),IHC results showed that 290 cases(84.3％)had proficient mismatch repair(pMMR),and 54 cases(15.7％)had deficient mismatch repair(dMMR).The results of PCR-CE showed that 53 patients(15.4％)had high microsatellite instability(MSI-H),and 291 patients(84.6％)had microsatellite stable(MSS).The consistency analysis of IHC and PCR-CE results showed that the Kappa value was 0.923(P＜0.001).Among the 168 cases of en-dometrial cancer(EC),IHC results showed that 42 cases(25.0％)had dMMR and 126 cases(75.0％)had pMMR;The results of PCR-CE showed that 39 cases(23.2％)had MSI-H and 129 cases(76.8％)had MSS.The consisten-cy analysis of the two methods showed that the Kappa value was 0.854(P＜0.001).Among the 165 cases of gastric cancer,IHC results showed that 22(13.3％)had dMMR and 143(86.7％)had pMMR;The results of PCR-CE showed that 22 cases(13.3％)had MSI-H and 143 cases(86.7％)had MSS/low microsatellite instability(MSI-L).The consistency analysis of IHC and PCR-CE results showed that the Kappa value was 0.843(P＜0.001).Among the 58 cases of head and neck squamous cell carcinoma(HNSCC),IHC results showed that 3 cases(5.2％)had dMMR and 55 cases(94.8％)had pMMR;The results of PCR-CE showed that 3 cases(5.2％)had MSI-H and 55 cases(94.8％)had MSS.The consistency analysis of the two methods showed that the Kappa value was 1.000(P＜0.001).Conclusion IHC and PCR-CE have high consistency and reliability in detecting MSI status in patients with CRC,EC,gastric cancer and HNSCC,and their combined application is helpful to improve the accuracy of MSI detec-tion in patients with solid tumors.

Objective To understand the current situation of healthcare-associated infection(HAI)in China,pro-vide data support and decision-making basis for formulating scientific and effective strategies for HAI prevention and control.Methods A nationwide cross-sectional survey on HAI was conducted among various types and levels of medical institutions in China according to a unified protocol of bedside surveys and case investigations.Results In 2024,a total of 5 736 medical institutions and 2 751 765 patients were surveyed.Among them,34 889 HAI cases were identified,with a prevalence rate of 1.27％.The number of HAI episodes was 38 032,and case prevalence rate was 1.38％.The prevalence rate of HAI in medical institutions in different regions of China ranged from 0.66％to 2.35％.Among medical institutions of different scales,those with a bed capacity of ≥900 had the high-est incidence of HAI,reaching 1.65％.The most common infection site was the lower respiratory tract(44.66％),followed by the urinary tract(12.94％),surgical site(9.32％),upper respiratory tract(7.02％),and bloodstream infection(5.78％).The top 3 departments with the highest HAI rates were the general intensive care unit(10.02％),department of neurosurgery(5.51％),and department(group)of hematology(5.34％).A total of 23 238 strains of HAI pathogens were detected,with 10 714 strains(46.10％)from lower respiratory tract speci-mens.The top 5 detected strains were Klebsiella pneumoniae(14.76％),Pseudomonas aeruginosa(13.33％),Escherichia coli(12.79％),Acinetobacter baumannii(9.23％),and Staphylococcus aureus(7.88％).231 944 pa-tients underwent class Ⅰ incision surgery were monitored,with 1 647 cases experienced surgical site infection,and the prevalence rate of surgical site infection was 0.71％.The number of patients who should undergo pathogen de-tection(patients receiving therapeutic and therapeutic combined prophylactic antimicrobial agents)was 715 179,while the actual number was 480 492,with a pathogen detection rate of 67.18％.425 225 patients received patho-genic detection before treatment,with a detection rate of 59.46％.Conclusion The overall HAI prevalence in Chi-na is lower,showing disparities among medical institutions of different regions and scales.Therefore,precise imple-mentation of measures is necessary for HAI prevention and control,with a focus on high-risk institutions and high-risk departments,key areas,and critical procedures.All levels of medical institutions should continuously reduce the incidence of HAI by strengthening monitoring,standardizing the use of antimicrobial agents,and reinforcing basic HAI prevention and control measures.

Objective:To investigate the impact of endocrine therapy(ET)on the prognosis of breast cancer patients with low estrogen re-ceptor(ER)expression and to identify the associated independent prognostic factors.Methods:We retrospectively analyzed the clinicopath-ological data of 387 female patients with breast cancer with low ER expression and at stage Ⅰ-Ⅲ TNM who were treated surgically at Xinji-ang Medical University Affiliated Tumor Hospital between January 2014 and May 2020.The patients were stratified into ET and non-ET groups based on whether they received ET administration.Propensity score matching(PSM)was used to balance intergroup differences,and survival analysis along with Cox proportional hazards models were applied to assess prognosis and the associated independent influencing factors.Results:Among the 387 patients,214(55.3%)received ET and 173(44.7%)did not.Baseline data indicated that the non-ET group exhibited higher proportions of ethnic minorities and patients with regional lymph node involvement,advanced TNM stage,and no radio-therapy administration.After PSM,baseline characteristics showed no significant differences between the ET(n=132)and non-ET groups(n=132).Pre-PSM survival analysis revealed 5-year overall survival(OS)rates of 93.4%and 72.8%and 5-year disease-free survival(DFS)rates of 74.1%and 60.9%for the ET and non-ET groups,respectively;both rates showed statistically significant differences between the groups(P<0.05).Post-PSM,the 5-year OS rates significantly differed between the ET and non-ET groups(91.6%and 80.1%,respectively;P<0.05),whereas no significant difference in 5-year DFS rates were observed(71.1%[ET group]vs.68.1%[non-ET group],P>0.05).Multivariate ana-lysis identified the following independent prognostic factors:for OS,ET administration,CK5/6 status,tumor size T3,lymph node status N2/N3,and radiotherapy administration(all P<0.05).For DFS,the independent factors included CK5/6 status,tumor size T3,lymph node status N2/N3,radiotherapy administration,and targeted therapy(all P<0.05).Conclusions:Endocrine therapy significantly improved the 5-year OS in breast cancer patients with low ER expression.CK5/6 positivity,large tumor size(T3),and regional lymph node involvement(N2/N3)were independent risk factors for both 5-year OS and DFS.Adjuvant radiotherapy and trastuzumab-based therapy significantly im-proved prognosis.

Objective To evaluate the efficacy of Compound Fipronil Spot-on Solution in repelling canine ticks.Methods A total of 140 dogs infested with ticks were randomly selected from regions in southern and northern China and assigned to four groups:southern test drug group,southern control drug group,northern test drug group,and northern control drug group.Each group comprised 35 dogs.Each dog was administered the prescribed dose.The number of ticks was counted on days 1,7,14,21,and 28 following the administration.The negative conversion and average reduction rates of the tick population were then subjected to statistical analyse.Results The mean efficacy of the test drug was 100%in both the southern and northern cohorts,28 days post-treatment.The control drug showed comparable efficacy,reaching a mean reduction of 100%in both regions by the same time point.No additional clinical manifestations or adverse events were observed across all treated dogs.Conclusions Compound Fipronil Spot-on Solutions effectively treats and prevents ticks in dogs in different regions of China.A single dose remains effective for up to 28 days,thus providing a convenient,effective solution.