The signaling mechanisms and developmental dynamics that govern the divergence of myocardial and epicardial lineages during human heart development remain poorly understood. Here, we developed a human pluripotent stem cell-based cardiac development model and employed time-course single-cell RNA sequencing to delineate cardiac lineage specification trajectories. We identified retinoic acid (RA) as a critical fate switch at the cardiac mesoderm stage. RA instructs epicardial lineage commitment of cardiac mesoderm through a primed-epicardium to proepicardium-like population and finally to epicardium, a process requiring precise BMP modulation. Conversely, RA absence directs cardiac mesoderm along a default myocardial pathway, yielding developing and mature cardiomyocytes. Both trajectories are governed by the hierarchical activation of key transcription factors. Our study integrates signaling and dynamics to elucidate the temporal regulatory network of the RA-BMP axis in human cardiac fate determination. These findings provide fundamental insights into human cardiogenesis and a crucial roadmap for modeling heart disease and advancing regenerative strategies.

Endometrial polyps (EPs) are common uterine lesions associated with abnormal uterine bleeding and infertility, yet their pathogenesis remains poorly defined. Here, we performed single-cell RNA sequencing of normal endometrium, para-polyp, and polyp tissues, identifying distinct cellular compositions and transcriptional programs. EPs showed enhanced estrogen signaling and increased epithelial proliferation, accompanied by decreased expression of cytokines and reduced T cell cytotoxicity. Notably, we observed epithelial subpopulations with elevated copy number variations and transcription factors associated with hyperplasia. Cell-cell communication analyzes revealed aberrant stromal-epithelial crosstalk, characterized by upregulated WNT, IGF, and VEGF signaling originating from stromal cells. Spatial transcriptomic analyzes further demonstrated enhanced WNT signaling between stromal and epithelial compartments in endometrial cancer. In vitro glandular organoid models showed that epithelial transcriptional alterations contribute to polyp formation. These findings highlight a critical role of stromal-epithelial interactions in EP development and suggest potential therapeutic targets.

Vibrio parahaemolyticus is a leading cause of seafood-associated gastroenteritis, posing increasing public health challenges globally. We conducted a large-scale molecular epidemiological analysis of 7932 clinical isolates collected from seafood-associated diarrheal patients in 69 sentinel hospitals across 11 prefecture-level cities in Zhejiang Province between 2016 and 2023. Whole-genome sequencing (WGS) was performed on 1261 representative isolates, supplemented by 11,555 publicly available genomes. We analyzed temporal serotype dynamics, antimicrobial resistance genes (ARGs), virulence factors, and pangenomic diversity, with an emphasis on the globally prevalent ST3 lineage. Notably, the O10:K4 serotype emerged as the second most common within ST3, increasing in prevalence from 40.0 % to 65.6 % over the study period. O10:K4 isolates exhibited a significantly higher frequency of tdh (96.3 % vs. 78.2 %) and complete T3SS2 effector gene clusters. Nearly all isolates harbored tet(35) and blaCARB-22, indicating widespread resistance. Comparative pangenome analysis revealed O10:K4-specific acquisitions in chemotaxis and LPS synthesis (e.g., per, mcpQ, fcl) and loss of key polyamine metabolism genes (aguA, puu operon). These findings suggest that the emergence and clonal expansion of the O10:K4 serotype represent a significant epidemiological shift characterized by enhanced virulence and multidrug resistance. Sustained genomic surveillance is imperative to guide seafood safety management and clinical interventions.

Ischemic acute kidney injury may accelerate the progression to end-stage renal disease. Megalin has been shown to shuttle stanniocalcin (STC)-1 (which promotes mitochondrial antioxidant defenses) to the mitochondria through the retrograde-early endosomes-to-Golgi pathway; knockout of megalin in cultured cells has been reported to impair glycolysis and mitochondrial respiration. This study sought to determine kidney phenotype after ischemia/reperfusion (I/R) kidney injury in mice with tubular epithelium-specific deletion of megalin. Mice (on C57BL/6 background) with conditional tubular epithelium-specific knockout of megalin (tLrp2KO) and mice with combined conditional tubular epithelium-specific knockout of megalin and overexpression of STC1 (tLrp2KO;tSTC1O) were subjected to ischemia (clamping of renal pedicles), followed by reperfusion for 1, 3, 10, 45, and 90 days. Serum creatinine was measured and kidneys were harvested for analysis. After I/R and compared with control mice, tLrp2KO mice displayed worse acute kidney injury, severe and persistent inflammation, diminished tubular epithelial cell proliferation, up-regulation of TGFβ1 signaling, fibrosis, and accelerated progression to chronic kidney disease. Kidney injury was not rescued in tLrp2KO;tSTC1O mice, consistent with megalin-dependent renal protection by STC1. Freshly isolated proximal tubule fragments from tLrp2KO mice or cultured proximal tubule epithelial cells with megalin knockout displayed activation of TGFβ1 signaling, consistent with modulation of TGFβ1 signaling by megalin. In conclusion, tubular epithelium-specific deletion of megalin aggravates I/R kidney injury, up-regulates TGFβ1 signaling, and accelerates chronic kidney disease progression.

Recent advances in human embryo culture and revised guidelines from the International Society for Stem Cell Research (ISSCR) give grounds for extending the 14-day limit. To prepare for any possible future change in China’s regulation, we propose a path toward cautious extension with strict oversight, balancing scientific progress with ethical responsibility.

Migratory birds play a pivotal role in the global dissemination of antimicrobial resistance genes (ARGs), with shorebirds relying on coastal wetlands during their long-distance migrations, environments often contaminated and conducive to ARG transmission. However, systematic investigations into antimicrobial resistance (AMR) in shorebirds remain scarce. During spring and autumn of 2023, we collected 893 throat and cloacal swabs from 480 shorebirds, representing 28 species, at Chongming Dongtan, a critical stopover along the East Asian-Australasian Flyway. Our analysis identified six strains, including four extra-intestinal pathogenic E. coli (ExPEC) and two K. pneumoniae, that exhibited resistance to third-generation cephalosporins, with three ExPEC strains exhibiting significant virulence in Galleria mellonella infection assays. We identified two conjugative plasmids: E042113F_p1, carrying the blaCMY-2 gene in E. coli, and M50_p2, carrying the blaKPC-2 gene in a hypervirulent K. pneumoniae with a virulence plasmid harboring the aerobactin system. Bioinformatic and experimental analyses confirmed that these plasmids could transfer without any fitness cost, remaining stable for at least 30 passages. Surprisingly, genomic tracing revealed that among the plasmids similar to E042113F_p1 (blaCMY-2), the earliest was identified in a Chinese swallow in 2015, with subsequent detections in wild birds from Mongolia (2017), Russia (2018), and Australia (2019). Notably, these E04-CMY-like/M50-KPC-like plasmids predominantly originated from human sources, underscoring the pivotal role of human activity in the cross-species transmission of AMR. This human-mediated transmission of resistance elements into wildlife posed a substantial risk for amplifying and disseminating AMR through long-range migratory bird movements, highlighting the urgent need for international collaboration under a One Health framework. Integrated surveillance, environmental management, and stringent antibiotic stewardship are critical to mitigating the risks posed by migratory birds in amplifying and spreading AMR across ecosystems.

IntroductionClostridioides difficile Infection (CDI) is more prevalent in people with hematologic diseases. However, epidemiological characteristics are poorly understood.MethodsFrom July 2016 to November 2021, we studied the epidemiology of CDI in patients with hematological diseases at a tertiary teaching hospital in Shanghai, China.ResultsIn hematological patients, the prevalence of CDI was 21.6%, with 89.8% hospital-acquired infections. C. difficile ST81, which is a multidrug-resistant strain carrying only the toxin B, is the most common strain (38.1%), followed by ST3 (16.7%) and ST2 (9.5%). Clindamycin and moxifloxacin resistance rates of all C. difficile species were 64.3% and 31%, respectively, and no isolate was resistant to vancomycin, linezolid, metronidazole, teicoplanin, or daptomycin. DiscussionThis study provides a comprehensive characterization of CDI in hematological patients, highlighting the urgent need for enhanced surveillance and preventive strategies against this emerging nosocomial threat.

ObjectiveTo conduct molecular prevalence and genetic polymorphism analysis of 24 Swine Farm associated C. difficile ST11 strains, in addition to other representative sequenced ST strains.MethodsThe collected C. difficile strains underwent whole genome sequencing and bioinformatic analysis using the illumina NovaSeq platform, SPAdes, Prokka, MOB-suite, and FastTree. Virulence and antibiotic resistance genes were identified through NCBI Pathogen Database. Cytotoxicity tests were conducted on HT-29 cells and Vero cells to verify the function of toxin A and toxin B.ResultsThe most prevalent resistance genes in ST11 were found to be against β-lactamases, aminoglycosides, and tetracycline. A C. difficile isolate (strain 27) with tcdA deletion and high antibiotic resistance genes was far apart from other swine farm associated ST11 isolates in the phylogenetic branch. The remarkable genetic similarity between animal and human C. difficile strains suggests potential transmission of ST11 strains between animals and humans. The plasmid replicon sequences repUS43 were identified in all ST11 strains except one variant (strain 27), and 91.67% (22/24) of these were assessed by MOB-typer as having mobilizable plasmids.ConclusionSwine farm associated C. difficile ST11 carried fewer virulence genes than ST11 strains collected from NCBI database. It is critical to monitor the evolution of C. difficile strains to understand their changing characteristics, host-switching, and develop effective control and prevention strategies.

IntroductionAutism spectrum disorder (ASD) is a multifaceted developmental condition that commonly appears during early childhood. The etiology of ASD remains multifactorial and not yet fully understood. The identification of biomarkers may provide insights into the underlying mechanisms and pathophysiology of the disorder. The present study aimed to explore the causes of ASD by investigating the key biomedical markers, trace elements, and microbiota factors between children with autism spectrum disorder (ASD) and control subjects.MethodsMedline, PubMed, ProQuest, EMBASE, Cochrane Library, PsycINFO, Web of Science, and EMBSCO databases have been searched for publications from 2012 to 2023 with no language restrictions using the population, intervention, control, and outcome (PICO) approach. Keywords including “autism spectrum disorder,” “oxytocin,” “GABA,” “Serotonin,” “CRP,” “IL-6,” “Fe,” “Zn,” “Cu,” and “gut microbiota” were used for the search. The Joanna Briggs Institute (JBI) critical appraisal checklist was used to assess the article quality, and a random model was used to assess the mean difference and standardized difference between ASD and the control group in all biomedical markers, trace elements, and microbiota factors.ResultsFrom 76,217 records, 43 studies met the inclusion and exclusion criteria and were included in this meta-analysis. The pooled analyses showed that children with ASD had significantly lower levels of oxytocin (mean differences, MD = −45.691, 95% confidence interval, CI: −61.667, −29.717), iron (MD = −3.203, 95% CI: −4.891, −1.514), and zinc (MD = −6.707, 95% CI: −12.691, −0.722), lower relative abundance of Bifidobacterium (MD = −1.321, 95% CI: −2.403, −0.238) and Parabacteroides (MD = −0.081, 95% CI: −0.148, −0.013), higher levels of c-reactive protein, CRP (MD = 0.401, 95% CI: 0.036, 0.772), and GABA (MD = 0.115, 95% CI: 0.045, 0.186), and higher relative abundance of Bacteroides (MD = 1.386, 95% CI: 0.717, 2.055) and Clostridium (MD = 0.281, 95% CI: 0.035, 0.526) when compared with controls. The results of the overall analyses were stable after performing the sensitivity analyses. Additionally, no substantial publication bias was observed among the studies.InterpretationChildren with ASD have significantly higher levels of CRP and GABA, lower levels of oxytocin, iron, and zinc, lower relative abundance of Bifidobacterium and Parabacteroides, and higher relative abundance of Faecalibacterium, Bacteroides, and Clostridium when compared with controls. These results suggest that these indicators may be a potential biomarker panel for the diagnosis or determining therapeutic targets of ASD. Furthermore, large, sample-based, and randomized controlled trials are needed to confirm these results.

Carbapenem-resistant Escherichia coli (CREC) poses a severe global public health risk. This study reveals the worldwide geographic spreading patterns and spatiotemporal distribution characteristics of resistance genes in 7918 CREC isolates belonging to 497 sequence types (ST) and originating from 75 countries. In the last decade, there has been a transition in the prevailing STs from highly virulent ST131 and ST38 to higher antibiotic-resistant ST410 and ST167. The rise of multi-drug resistant strains of CREC carrying plasmids with extended-spectrum beta-lactamase (ESBL) resistance genes could be attributed to three important instances of host-switching events. The spread of CREC was associated with the changing trends in blaNDM-5, blaKPC-2, and blaOXA-48, as well as the plasmids IncFI, IncFII, and IncI. There were intercontinental geographic transfers of major CREC strains. Various crucial transmission hubs and patterns have been identified for ST131 in the United Kingdom, Italy, the United States, and China, ST167 in India, France, Egypt, and the United States, and ST410 in Thailand, Israel, the United Kingdom, France, and the United States. This work is valuable in managing CREC infections and preventing CREC occurrence and transmission inside healthcare settings and among diverse hosts.

Staphylococcus aureus (S. aureus) pneumonia has become an increasingly important public health problem. Recent evidence suggests that epigenetic modifications are critical in the host immune defence against pathogen infection. In this study, we found that S. aureus infection induces the expression of histone deacetylase 6 (HDAC6) in a dose-dependent manner. Furthermore, by using a S. aureus pneumonia mouse model, we showed that the HDAC6 inhibitor, tubastatin A, demonstrates a protective effect in S. aureus pneumonia, decreasing the mortality and destruction of lung architecture, reducing the bacterial burden in the lungs and inhibiting inflammatory responses. Mechanistic studies in primary bone marrow-derived macrophages demonstrated that the HDAC6 inhibitors, tubastatin A and tubacin, reduced the intracellular bacterial load by promoting bacterial clearance rather than regulating phagocytosis. Finally, N-acetyl-L- cysteine, a widely used reactive oxygen species (ROS) scavenger, antagonized ROS production and significantly inhibited tubastatin A-induced S. aureus clearance. These findings demonstrate that HDAC6 inhibitors promote the bactericidal activity of macrophages by inducing ROS, an important host factor for S. aureus clearance and production. Our study identified HDAC6 as a suitable epigenetic modification target for preventing S. aureus infection, and tubastatin A as a useful compound in treating S. aureus pneumonia.

While accumulated publications support the existence of neurogenesis in the adult human hippocampus, the homeostasis and developmental potentials of neural stem cells (NSCs) under different contexts remain unclear. Based on our generated single-nucleus atlas of the human hippocampus across neonatal, adult, aging, and injury, we dissected the molecular heterogeneity and transcriptional dynamics of human hippocampal NSCs under different contexts. We further identified new specific neurogenic lineage markers that overcome the lack of specificity found in some well-known markers. Based on developmental trajectory and molecular signatures, we found that a subset of NSCs exhibit quiescent properties after birth, and most NSCs become deep quiescence during aging. Furthermore, certain deep quiescent NSCs are reactivated following stroke injury. Together, our findings provide valuable insights into the development, aging, and reactivation of the human hippocampal NSCs, and help to explain why adult hippocampal neurogenesis is infrequently observed in humans.

To improve the understanding of antimicrobial resistance (AMR) in migratory birds derived-Enterobacter cloacae (E. cloacae) complex and its spread at the human-bird-environment interface, we isolated 11 strains of third-generation cephalosporin-resistant E. cloacae from 1003 specimens from 29 migratory bird species over two years in Chongming, Shanghai, China. The comprehensive analysis incorporated second- and third-generation sequencing techniques and extensive bioinformatic analysis. Four human-associated E. cloacae sequence types (STs), including ST432, ST412, ST1, and ST639, were found in migratory birds. We confirmed that the blaCMH-4 and blaCMH-6 genes were the major genotypes of the β-lactamase resistance genes in E. cloacae found in migratory birds. In addition, a thorough genomic analysis was performed on a global collection of 398 E. cloacae isolates carrying the blaCMH gene from 46 different countries. China had the highest proportion with 19.10 % (76/398), followed by Singapore with 18.34 % (73/398), Nigeria with 15.83 % (63/398), and the USA with 14.07 % (56/398). The first transmission of E. cloacae carrying blaCMH-4 and blaCMH-6 was defined around 1894 and 1549, respectively. Time-based phylogenetic analysis revealed that host jumps among humans, birds, and the environment led to the emergence of modern strains with ESBL- and carbapenem-resistant genes from about 2004 to 2016. The detection rate of insertion sequences (IS) of E. cloacae carrying blaCMH from human sources is higher than that from migratory bird sources, which is related to the different genetic environments caused by antibiotic selective pressure. It is crucial to have a comprehensive understanding of the characteristics exhibited by blaCMH producing E. cloacae in different ecological environments. Our results contribute to the effective monitoring and implementation of proactive strategies to reduce the spread of multidrug-resistant E. cloacae.

Hormone-activated proteolysis is a recurring theme of plant hormone signaling mechanisms. In strigolactone signaling, the enzyme receptor DWARF14 (D14) and an F-box protein, MORE AXILLARY GROWTH2 (MAX2), mark SUPPRESSOR OF MAX2 1-LIKE (SMXL) family proteins SMXL6, SMXL7, and SMXL8 for rapid degradation. Removal of these transcriptional corepressors initiates downstream growth responses. The homologous proteins SMXL3, SMXL4, and SMXL5, however, are resistant to MAX2-mediated degradation. We discovered that the smxl4 smxl5 mutant has enhanced responses to strigolactone. SMXL5 attenuates strigolactone signaling by interfering with AtD14-SMXL7 interactions. SMXL5 interacts with AtD14 and SMXL7, providing two possible ways to inhibit SMXL7 degradation. SMXL5 function is partially dependent on an ethylene-responsive-element binding-factor-associated amphiphilic repression (EAR) motif, which typically mediates interactions with the TOPLESS family of transcriptional corepressors. However, we found that loss of the EAR motif reduces SMXL5-SMXL7 interactions and the attenuation of strigolactone signaling by SMXL5. We hypothesize that integration of SMXL5 into heteromeric SMXL complexes reduces the susceptibility of SMXL6/7/8 proteins to strigolactone-activated degradation and that the EAR motif promotes the formation or stability of these complexes. This mechanism may provide a way to spatially or temporally fine-tune strigolactone signaling through the regulation of SMXL5 expression or translation.

BACKGROUND:Based on its objective characteristics, laboratory markers have always been the research direction of clinical diagnosis and assessment of mental disorders including Alzheimer's disease.METHODS:MTT Colorimetric Assay, ELISA, and quantitative PCR were used to investigate the responsiveness of peripheral blood mononuclear cells (PBMCs) to mitogen Lipopolysaccharides (LPS) and Phytohemagglutinin (PHA), PBMCs genomic methylation and hydroxymethylation levels, nuclear DNA and mitochondrial DNA damage, respiratory chain enzyme activities, and circulating cell-free mitochondrial DNA levels were detected in 90 patients with Alzheimer's disease.RESULTS:In the Alzheimer's disease group, LPS stimulated PBMCs viability, TNF-α secretion, PHA stimulated IL-10 secretion, genomic DNA methylation levels, circulating cell-free mitochondrial DNA copies, citrate synthase activity were reduced compared to the control; while the LPS stimulated PBMCs IL-1α secretion, PHA stimulated IL-1α and IFN-γ secretion, plasma IL-6 and TNF-α, mitochondrial DNA damages were increased compared to the control.CONCLUSIONS:The reactivity of peripheral blood mononuclear cells to mitogens, mitochondrial DNA integrity characteristics, and cell-free mitochondrial DNA copies may be used as candidate laboratory biomarkers to help clinical management of Alzheimer's disease.

抗微生物药物耐药性(antimicrobial resistance,AMR)是一个跨地区、跨领域和跨学科问题,涉及临床医学、动物医学、环境健康学及政策研究等,全球各地均面临着严峻的AMR局面.随着生物技术的发展,宏基因组二代测序技术(metagenomic next generation sequencing,mNGS)的出现为抗微生物药物耐药基因(an-tibiotic resistance genes,ARGs)的研究带来了巨大突破和转机.本文阐述了研究AMR的重要性和mNGS技术的发展现状,分别总结了 mNGS技术在人、动物和环境领域ARGs研究的应用现状,并对基于宏基因组测序的ARGs在人—动物—环境界面的多生境和多层面研究进行了展望.

在检验医学课程的开展中,以医学院教师的理论教学为依托,引入校企联合培养模式,在专业课程的开展中加入企业教师进课堂和专业学生入企业,结合免疫读书报告会、实验操作技能大赛、大学生创新计划和毕业论文课题等由企业技术教师参与作为技术指导、评审监督、奖学金支持的项目,追踪近5年各项目的实践成绩,评估校企联合培养模式对于培养新时代卓越临床检验工作者的价值.校企联合培养模式能使学生的专业水平、科学素养和就业竞争力全面提升,为培养卓越的医学创新技术人才做贡献.

With increasing attention paid to the concept of the microbiota–gut–brain axis, mounting evidence reveals that the gut microbiota is involved in a variety of neurological and psychiatric diseases. However, gut microbiota changes in major depressive disorder (MDD) patients and their association with disease mechanisms remain undefined. Fifty MDD patients and sixty healthy controls were recruited from the Shanghai Healthy Mental Center, China. Fecal samples were collected, and the compositional characteristics of the intestinal flora were determined in MDD patients by MiSeq sequencing. Venous blood was collected for the detection of plasma indoleamine-2,3-dioxygenase (Ido), kynurenine (Kyn) and tryptophan (Trp) levels. Stool samples of bacterial 16S sequencing was carried out. A total of 2,705,809 optimized sequences were obtained, with an average of 54,116 per sample. More unique OTUs were observed at the family, genus and species levels in the control group compared with the MDD cases. Further analysis showed significant changes in the α- and β-diversities and relative abundance levels of gut microbial entities in MDD patients, as well as elevated amounts of Ido and Kyn indicating Kyn pathway activation, KEGG bacterial 16S function prediction analysis shows a variety of amino acids and metabolic (including Ido, Trp and Kyn) changes in the body of patients with MDD. These may result in increased neurotoxic metabolites and reduced generation of serotonin in the disease process. These changed factors may potentially be utilized as biomarkers for MDD in the future, playing more important roles in the disease course.

目的 对疫情期间临床医学生实验诊断学线上课程进行教学成效的调查与分析,为实验诊断学在线课程的优化和实现线上教学与线下教学同质等效目标提供理论依据.方法 选择上海交通大学医学院 2020级临床医学专业学生 79 名作为研究对象,其中临床医学五年制(儿科班)学生 54 名,4+4(硕博班)学生 25 名.另选择接受线下教学的 2019 级临床医学五年制(儿科班)学生 56 名和 4+4(硕博班)学生 25 名作为对照.通过客观成绩对比分析、主观感受调查问卷两方面对学情、学习质量和教学满意度进行评估和反馈.结果 2020级儿科班学生实验诊断学成绩高于 2019 级,但差异无统计学意义(P>0.05);2020 级硕博班学生实验诊断学成绩较 2019 级有所下降,差异有统计学意义(P<0.05).44.4％的学生对实验诊断学线上教学表示非常满意,30.2％的学生表示比较满意.与线下教学比较,线上教学最重要的优势是能反复回放观看,有利于巩固知识点,但线上课程最大的缺点为缺少学习氛围,学生的学习积极性不高.结论 疫情期间实验诊断学线上课程取得了较好的效果.如何优化课程体系、提高学生线上学习积极性、促进线上线下教学的深度融合、提升线上教学质量是今后努力的方向.