The spatio-temporal heterogeneity of breast cell subsets forms the fundamental biological basis for physiological development and pathological progression, including tumorigenesis; however, its complex regulatory mechanisms are not yet fully elucidated. With its high-resolution capabilities, single-cell RNA sequencing (scRNA-seq) technology offers a powerful tool for dissecting this cellular heterogeneity. This technology enables the construction of high-precision breast cell atlases, the accurate identification of distinct cell subsets, and the reconstruction of differentiation trajectories from stem/progenitor cells to functional epithelial cells. By resolving the transcriptional regulatory networks that govern cell fate determination, intercellular communication patterns, and dynamic microenvironmental interactions, scRNA-seq has unveiled the molecular foundations of breast development and provided new perspectives on the pathogenesis of related diseases such as breast cancer and macromastia. Furthermore, scRNA-seq demonstrates significant potential for discovering early molecular markers of disease, deciphering tumor heterogeneity, and elucidating mechanisms of therapeutic resistance. The continued application of scRNA-seq for dissecting breast cell heterogeneity, combined with its integration with multi-modal data such as spatial omics, promises to provide critical evidence and new insights for revealing the molecular mechanisms of breast development-related diseases and for formulating precision therapeutic strategies.
Humans ; Single-Cell Analysis/methods* ; Female ; Breast Neoplasms/pathology* ; Sequence Analysis, RNA/methods* ; Breast/cytology*

Poly(ADP-ribosyl)ation (PARylation) is a specific form of post-translational modification (PTM) predominantly triggered by the activation of poly-ADP-ribose polymerase 1 (PARP1). However, the role and mechanism of PARylation in the advancement of acute kidney injury (AKI) remain undetermined. Here, we demonstrated the significant upregulation of PARP1 and its associated PARylation in murine models of AKI, consistent with renal biopsy findings in patients with AKI. This elevation in PARP1 expression might be attributed to trimethylation of histone H3 lysine 4 (H3K4me3). Furthermore, a reduction in PARylation levels mitigated renal dysfunction in the AKI mouse models. Mechanistically, liquid chromatography-mass spectrometry indicated that PARylation mainly occurred in receptor for activated C kinase 1 (RACK1), thereby facilitating its subsequent phosphorylation. Moreover, the phosphorylation of RACK1 enhanced its dimerization and accelerated the ubiquitination-mediated hypoxia inducible factor-1α (HIF-1α) degradation, thereby exacerbating kidney injury. Additionally, we identified a PARP1 proteolysis-targeting chimera (PROTAC), A19, as a PARP1 degrader that demonstrated superior protective effects against renal injury compared with PJ34, a previously identified PARP1 inhibitor. Collectively, both genetic and drug-based inhibition of PARylation mitigated kidney injury, indicating that the PARylated RACK1/HIF-1α axis could be a promising therapeutic target for AKI treatment.

Extensive epigenetic reprogramming involves in memory CD8+ T-cell differentiation. The elaborate epigenetic rewiring underlying the heterogeneous functional states of CD8+ T cells remains hidden. Here, we profile single-cell chromatin accessibility and map enhancer-promoter interactomes to characterize the differentiation trajectory of memory CD8+ T cells. We reveal that under distinct epigenetic regulations, the early activated CD8+ T cells divergently originated for short-lived effector and memory precursor effector cells. We also uncover a defined epigenetic rewiring leading to the conversion from effector memory to central memory cells during memory formation. Additionally, we illustrate chromatin regulatory mechanisms underlying long-lasting versus transient transcription regulation during memory differentiation. Finally, we confirm the essential roles of Sox4 and Nrf2 in developing memory precursor effector and effector memory cells, respectively, and validate cell state-specific enhancers in regulating Il7r using CRISPR-Cas9. Our data pave the way for understanding the mechanism underlying epigenetic memory formation in CD8+ T-cell differentiation.
CD8-Positive T-Lymphocytes/metabolism* ; Cell Differentiation ; Chromatin/immunology* ; Animals ; Mice ; Immunologic Memory ; Epigenesis, Genetic ; SOXC Transcription Factors/immunology* ; NF-E2-Related Factor 2/immunology* ; Mice, Inbred C57BL ; Gene Regulatory Networks ; Enhancer Elements, Genetic

Objective:To explore the feasibility of transvaginal core needle biopsy for pelvic masses under finger guidance during a vagino-recto-abdominal examination.Methods:The clinicopathological data and follow-up information of 29 patients with pelvic masses who underwent transvaginal core needle biopsy under finger guidance during a vagino-recto-abdominal examination at Affiliated Hospital of Southwest Medical University from January 2020 to July 2024 were collected, and the safety and diagnostic accuracy of the procedure were retrospectively analyzed.Results:(1) A total of 29 patients with pelvic masses were enrolled in this study, with a median age of 50 years (range: 29-73 years), and a median tumor diameter of 3.9 cm (range: 2.7-13.3 cm). Among these patients, 7 were newly diagnosed, and 22 were follow-up. The pre-procedure disease types included 21 patients (72%, 21/29) cervical cancer, 6 patients (21%, 6/29) epithelial ovarian cancer, and 2 patients (7%, 2/29) other suspected gynecologic tumors. (2) Among 29 patients with pelvic masses, 8 cases (28%, 8/29) were diagnosed with benign diseases according to core needle biopsy pathological findings, and 1 case suggested possible residual cervical cancer in the parametrial region by contrast-enhanced magnetic resonance imaging after radical chemoradiotherapy 3 months, while the result of core needle biopsy for this patient was negative, with follow-up after 1 year revealed progression of the lesion in the right parametrial area. Another patient underwent fine-needle aspiration cytology, which suggested gastrointestinal stromal tumor, requiring differentiation from endometriosis, and core needle biopsy pathology confirmed endometriosis, with follow-up at 6 months revealed no evidence of malignancy in this patient. The remaining 6 patients with benign diagnoses had follow-up periods exceeding 1 year without imaging or clinical evidence of local lesion progression or malignancy. Among the 21 patients (72%, 21/29) diagnosed with malignant tumors by core needle biopsy, 14 cases were suspected cases of residual or recurrent cervical cancer, 6 cases had advanced ovarian cancer, and 1 case had rectal cancer metastasis, with all biopsy diagnoses being consistent with preoperative clinical findings and imaging results. The overall diagnostic accuracy of the core needle biopsy was 97% (28/29). Among the 7 newly diagnosed patients, the diagnostic accuracy was 7/7, while it was 95% (21/22) for the 22 follow-up patients, with no statistically significant difference observed between the two groups ( P=1.000). (3) All 29 patients with pelvic masses successfully underwent transvaginal core needle biopsy guided by vagino-recto-abdominal examination. Among them, 28 cases (97%, 28/29) reported tolerable pain during the procedure, while 1 case (3%, 1/29) experienced transient syncope at the end of the procedure due to pain, which resolved within seconds. Vaginal bleeding exceeding 50 ml occurred in 3 patients (10%, 3/29) during paracervical tissue sampling, with the maximum blood loss being 150 ml, and hemorrhage was successfully controlled using vaginal tamponade. The overall incidence of adverse events during the core needle biopsy procedure was 14% (4/29). Conclusion:Transvaginal core needle biopsy for pelvic masses guided by vagino-recto-abdominal examination is a simple, safe, and accurate diagnostic method, suitable for patients with gynecologic malignancies, non-gynecologic malignancies suspected of pelvic mass metastasis, and other benign pelvic lesions.

Objective:To explore the feasibility of transvaginal core needle biopsy for pelvic masses under finger guidance during a vagino-recto-abdominal examination.Methods:The clinicopathological data and follow-up information of 29 patients with pelvic masses who underwent transvaginal core needle biopsy under finger guidance during a vagino-recto-abdominal examination at Affiliated Hospital of Southwest Medical University from January 2020 to July 2024 were collected, and the safety and diagnostic accuracy of the procedure were retrospectively analyzed.Results:(1) A total of 29 patients with pelvic masses were enrolled in this study, with a median age of 50 years (range: 29-73 years), and a median tumor diameter of 3.9 cm (range: 2.7-13.3 cm). Among these patients, 7 were newly diagnosed, and 22 were follow-up. The pre-procedure disease types included 21 patients (72%, 21/29) cervical cancer, 6 patients (21%, 6/29) epithelial ovarian cancer, and 2 patients (7%, 2/29) other suspected gynecologic tumors. (2) Among 29 patients with pelvic masses, 8 cases (28%, 8/29) were diagnosed with benign diseases according to core needle biopsy pathological findings, and 1 case suggested possible residual cervical cancer in the parametrial region by contrast-enhanced magnetic resonance imaging after radical chemoradiotherapy 3 months, while the result of core needle biopsy for this patient was negative, with follow-up after 1 year revealed progression of the lesion in the right parametrial area. Another patient underwent fine-needle aspiration cytology, which suggested gastrointestinal stromal tumor, requiring differentiation from endometriosis, and core needle biopsy pathology confirmed endometriosis, with follow-up at 6 months revealed no evidence of malignancy in this patient. The remaining 6 patients with benign diagnoses had follow-up periods exceeding 1 year without imaging or clinical evidence of local lesion progression or malignancy. Among the 21 patients (72%, 21/29) diagnosed with malignant tumors by core needle biopsy, 14 cases were suspected cases of residual or recurrent cervical cancer, 6 cases had advanced ovarian cancer, and 1 case had rectal cancer metastasis, with all biopsy diagnoses being consistent with preoperative clinical findings and imaging results. The overall diagnostic accuracy of the core needle biopsy was 97% (28/29). Among the 7 newly diagnosed patients, the diagnostic accuracy was 7/7, while it was 95% (21/22) for the 22 follow-up patients, with no statistically significant difference observed between the two groups ( P=1.000). (3) All 29 patients with pelvic masses successfully underwent transvaginal core needle biopsy guided by vagino-recto-abdominal examination. Among them, 28 cases (97%, 28/29) reported tolerable pain during the procedure, while 1 case (3%, 1/29) experienced transient syncope at the end of the procedure due to pain, which resolved within seconds. Vaginal bleeding exceeding 50 ml occurred in 3 patients (10%, 3/29) during paracervical tissue sampling, with the maximum blood loss being 150 ml, and hemorrhage was successfully controlled using vaginal tamponade. The overall incidence of adverse events during the core needle biopsy procedure was 14% (4/29). Conclusion:Transvaginal core needle biopsy for pelvic masses guided by vagino-recto-abdominal examination is a simple, safe, and accurate diagnostic method, suitable for patients with gynecologic malignancies, non-gynecologic malignancies suspected of pelvic mass metastasis, and other benign pelvic lesions.

Objective:To investigate the current status of standardized residency training (SRT) teaching activities at Zhongshan Hospital affiliated to Fudan University, and to explore optimization strategies.Methods:We collected behavioral data from the SRT course selection platform and resident questionnaire survey data throughout 2024. Descriptive analysis, chi-square test, Mann-Whitney U test, and multivariable logistic regression analysis were performed. Results:A total of 170 teaching sessions were conducted in 2024, with interactive practice-based sessions accounting for 42.35% and lecture-based sessions accounting for 47.06%. According to 536 questionnaires, residents' overall satisfaction with teaching activities scored 87.83 points (high satisfaction, ≥85 points; moderate satisfaction, <85 points). The proportion of high satisfaction with interactive practice-based sessions was significantly higher than that with lecture-based sessions (82.81% vs. 75.46%, P=0.034). The enrollment for weekday evening courses filled up significantly faster than that for weekday daytime courses [4 (2, 6) seconds vs. 12 (8, 15) seconds, P<0.001]. Interactive practice-based sessions [odds ratio ( OR)=1.6, 95% CI=1.1-2.3, P=0.018] and weekday evening sessions ( OR=1.4, 95% CI=1.0-2.0, P=0.048) significantly improved resident satisfaction. Conclusions:Optimizing course formats and scheduling can enhance the quality of SRT teaching activities.

Depression and circadian rhythms exhibit bidirectional interactions,suggesting a close association with the biological clock system.The biological clock in the suprachiasmatic nucleus of the hypothalamus is regulated by circadian genes.Recent clinical and basic research has revealed multifaceted associations between depression and circadian genes.For instance,significant phase abnormalities in BMAL1,PER2,and PER3 were detected in brain tissue from depressed patients,while plasma levels of CRY1,ARNTL,and PER1 proteins showed marked reduction,demonstrating good diagnostic value.Mice with CLOCK and BMAL1 knockouts exhibited depression-like behaviors.Single nucleotide polymorphisms(SNPs)in genes such as PER1 and PER3 directly influence depression susceptibility.Methylation levels of BMAL1,PER3,and CLOCK genes correlate closely with depressive symptoms.Antidepressant mechanisms like ketamine exert their effects by downregulating PER2 and other genes.This review summarizes the differential expression patterns of circadian clock genes in depression and associated therapeutic approaches,aiming to provide new theoretical foundations for precision diagnosis and personalized treatment strategies for depression.

Depression and circadian rhythms exhibit bidirectional interactions,suggesting a close association with the biological clock system.The biological clock in the suprachiasmatic nucleus of the hypothalamus is regulated by circadian genes.Recent clinical and basic research has revealed multifaceted associations between depression and circadian genes.For instance,significant phase abnormalities in BMAL1,PER2,and PER3 were detected in brain tissue from depressed patients,while plasma levels of CRY1,ARNTL,and PER1 proteins showed marked reduction,demonstrating good diagnostic value.Mice with CLOCK and BMAL1 knockouts exhibited depression-like behaviors.Single nucleotide polymorphisms(SNPs)in genes such as PER1 and PER3 directly influence depression susceptibility.Methylation levels of BMAL1,PER3,and CLOCK genes correlate closely with depressive symptoms.Antidepressant mechanisms like ketamine exert their effects by downregulating PER2 and other genes.This review summarizes the differential expression patterns of circadian clock genes in depression and associated therapeutic approaches,aiming to provide new theoretical foundations for precision diagnosis and personalized treatment strategies for depression.

Objective:To investigate the current status of standardized residency training (SRT) teaching activities at Zhongshan Hospital affiliated to Fudan University, and to explore optimization strategies.Methods:We collected behavioral data from the SRT course selection platform and resident questionnaire survey data throughout 2024. Descriptive analysis, chi-square test, Mann-Whitney U test, and multivariable logistic regression analysis were performed. Results:A total of 170 teaching sessions were conducted in 2024, with interactive practice-based sessions accounting for 42.35% and lecture-based sessions accounting for 47.06%. According to 536 questionnaires, residents' overall satisfaction with teaching activities scored 87.83 points (high satisfaction, ≥85 points; moderate satisfaction, <85 points). The proportion of high satisfaction with interactive practice-based sessions was significantly higher than that with lecture-based sessions (82.81% vs. 75.46%, P=0.034). The enrollment for weekday evening courses filled up significantly faster than that for weekday daytime courses [4 (2, 6) seconds vs. 12 (8, 15) seconds, P<0.001]. Interactive practice-based sessions [odds ratio ( OR)=1.6, 95% CI=1.1-2.3, P=0.018] and weekday evening sessions ( OR=1.4, 95% CI=1.0-2.0, P=0.048) significantly improved resident satisfaction. Conclusions:Optimizing course formats and scheduling can enhance the quality of SRT teaching activities.

Objective:To explore the genetic etiology of two patients with Gitelman syndrome (GS).Methods:Two patients who had presented at the Linyi People′s Hospital in January and June 2022 respectively were selected as the study subjects. Peripheral blood samples of them were collected and subjected to whole exome sequencing (WES). Electrolyte levels in their serum and urine were detected. Candidate variants were verified by Sanger sequencing. PyMOL software was used to predict the impact of the variants on the protein structure.Results:Patient 1 was a 27-year-old female with decreased serum levels of sodium, potassium, chloride and magnesium, along with decreased urine chloride and calcium. WES revealed that she has harbored compound heterozygous variants of the SLC12A3 gene, namely c. 1456G>A (p.D486N) and c. 179C>T (p.T60M). The former was inherited from her mother and known to be pathogenic. Patient 2 was a 4-year-old male with lower serum sodium, chloride and magnesium levels, and his serum potassium level was found to be critically low. He was found to harbor compound heterozygous variants of c. 602-16G>A and c. 805_806insTTGGCGTGGTCTCGGTCA (p.V268_T269insIGVVSV) of the SLC12A3 gene, which were inherited from his mother and father, respectively. Based on the guidelines from the American College of Medical Genetics and Genomics, both variants were predicted to be pathogenic (PVS1+ PM2_Supporting+ PP3; PVS1+ PM2_Supporting+ PM4). Conclusion:The above heterozygous variants of the SLC12A3 gene probably underlay the GS in these patients.