ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract（PAE） against human-derived lung adenocarcinoma organoids（LUAD-PDOs） and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid（Nor-PDOs） models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8（CCK-8） assay. Experimental groups included the model group（LUAD-PDOs）， normal group， model administration group（LUAD-PDOs+PAE）， and normal administration group（Nor-PDOs+PAE）. Hematoxylin-eosin（HE） staining was used to observe the pathological structures of PDOs， immunohistochemistry（IHC） was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7（CK-7） and Napsin A， TUNEL staining was applied to detect cell apoptosis. RNA sequencing（RNA-Seq） was conducted to identify differentially expressed genes（DEGs）， followed by Gene Ontology（GO）， Kyoto Encyclopedia of Genes and Genomes（KEGG）， and Gene Set Enrichment Analysis（GSEA）， alongside protein-protein interaction（PPI） network analysis to screen core mechanisms. Finally， key targets were validated by integrating external database analysis with immunofluorescence（IF）. ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L^-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures， whereas LUAD-PDOs displayed dense， solid structures. CCK-8 and TUNEL assays revealed that， compared with the model group， PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells， while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention（347 up-regulated and 372 down-regulated）（P<0.05）. GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism， including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway（P<0.05）. PPI network analysis indicated that breast cancer type 1 susceptibility protein（BRCA1） and checkpoint kinase 1（CHEK1） were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention（P<0.05）. Furthermore， survival analysis based on The Cancer Genome Atlas（TCGA） database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD（P<0.05）. ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis， its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway， with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.

ObjectiveFlavonoids are clinically significant natural products, yet their oxygen-glycosylation in aqueous environments relies heavily on expensive nucleotide-activated sugar donors such as UDP-glucose. Liquid-liquid phase separation (LLPS) creates specialized, membraneless physicochemical microenvironments capable of modulating enzymatic functions and overcoming mass transfer limitations. This study aims to investigate whether the gut microbiota-derived DgpB/C complex—a multienzyme system traditionally recognized for cleaving stable C-glycosidic bonds and facilitating isomerization—can undergo functional remodeling within phase-separated condensates. Our core objective is to elucidate the role of phase separation in expanding enzymatic catalytic plasticity and to provide a non-canonical, highly cost-effective biocatalytic mechanism for the direct utilization of free sugars in the synthesis of O-glycosylated natural products. MethodsAn artificial phase-separation platform was constructed utilizing the multivalent arginine-glycine-glycine motif (RGG)-repeat domain derived from the Caenorhabditis elegans LAF-1 protein. To ensure precise spatial compartmentalization, the DgpB/C complex was specifically recruited into the RGG condensates via a high-affinity SZ1/SZ2 heterodimerization tag system. Condensate formation and substrate partitioning were visualized using light and confocal fluorescence microscopy. The chemical structures and regioselectivity of the reaction products were rigorously characterized using high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS)/MS systems. Furthermore, molecular docking and 20-ns molecular dynamics (MD) simulations were performed via the Hermite platform and Uni-GBSA pipeline to elucidate the structural and thermodynamic basis underlying the phase-transition-induced functional shift. ResultsWe observed that the condensates formed by RGG proteins selectively recruited and significantly enriched hydrophobic flavonoid substrates. Strikingly, within the LLPS microenvironment, the DgpB/C complex—which typically exhibits only degradative or isomerase activities—underwent a profound functional remodeling, transforming into an efficientO-glycosyltransferase. Diverging from canonical pathways that require high-energy donors, the sequestered DgpB/C complex directly utilized unactivated free glucose to catalyze the formation of new O-glycosidic bonds. This remodeled activity was found to be evolutionarily conserved across DgpB/C homologs from diverse gut microbiota strains, such as P581a and W974-1. LC-MS/MS analysis further revealed that the phase-separated environment reduced the regioselectivity constraints of the enzyme, enabling catalytic action on multiple hydroxyl positions of the flavonoid scaffold. MD simulations further indicated that the low-water-activity microenvironment of the condensates reconfigured the conformational dynamics of the catalytic pocket, favoring a spatial orientation highly conducive to dehydration condensation. ConclusionThis study demonstrates that LLPS drives the functional remodeling of the gut microbiota enzyme DgpB/C through the reconfiguration of the physicochemical microenvironment. These findings challenge traditional perceptions regarding the functional boundaries of metabolic enzymes and reveal a novel biocatalytic pathway that bypasses the requirement for nucleotide-activated sugars. Consequently, this provides a highly promising artificial compartmentalization strategy for the green manufacturing of complex, high-value-added natural products.

Objective To analyze the factors affecting the prognosis of patients with locally advanced or metastatic non-small cell lung cancer (NSCLC) undergoing immunotherapy and construct an individualized prognostic nomogram prediction model. Methods A retrospective analysis was conducted on the clinical data of 385 patients with driver gene-negative, locally advanced or metastatic NSCLC who received first-line immune checkpoint inhibitors. Univariate and multivariate Cox regression analyses were used to identify prognostic risk factors, and a prognostic nomogram model was established. The predictive performance of the model was evaluated using the concordance index (C-index), time-dependent receiver operating characteristic (ROC) curves and area under the curve (AUC), and calibration curves. The cutoff value of the nomogram was calculated to stratify patients by risk. Survival curves were calculated by Kaplan-Meier analysis. Results Age (HR=1.775, 95%CI: 1.265-2.490), degree of differentiation (HR=0.365, 95%CI: 0.257-0.519), low PD-L1 expression (HR=0.661, 95%CI: 0.455-0.960), high PD-L1 expression (HR=0.423, 95%CI: 0.297-0.603), SCC-Ag (HR=1.549, 95%CI: 1.109-2.163), and CONUT score (HR=2.527, 95%CI: 1.797-3.554) were independent risk factors affecting overall survival (OS) of patients with NSCLC undergoing immunotherapy. The nomogram prediction model constructed on the basis of these factors had a C-index of 0.767. Time-dependent ROC curves for survival showed that the AUCs for 1-, 2-, and 3-year OS were 0.830, 0.853, and 0.886, respectively. Calibration curves indicated that the nomogram-predicted survival rates were in good agreement with the actual outcomes. The cutoff value for the study’s nomogram prediction model was 136.60 points, and survival curves showed statistically significant differences between different risk groups (P<0.05). Conclusion The nomogram model established in this study can effectively predict the prognosis of patients with driver gene-negative locally advanced or metastatic NSCLC treated with first-line immunosuppressive therapy. It provides a new tool for assessing prognosis and aids clinicians in formulating individualized treatment plans.

Objective To compare and analyze malaria epidemic trends and characteristics of initial confirmed cases before and after malaria eliminating in Qingjiangpu District, Huai′an City, and to provide a scientific basis for improving strategies to prevent imported malaria retransmission. Methods Malaria case data from Qingjiangpu District were obtained from the National Infectious Disease Information Reporting System for 2010-2023. Case investigation and epidemiological data were summarized and analyzed for the pre-elimination period (2010-2016)and post-elimination period(2017-2023). Results A total of 185 malaria cases were reported between 2010 and 2023, with one locally acquired case in both 2010 and 2011; the remainder were imported, primarily falciparum malaria. Post-elimination cases decreased from 140 to 45, showing a statistically significant difference in case distribution ratios(χ2=9.642, P<0.05). Seasonal peaks occurred in early January and mid-July before elimination but showed no distinct pattern afterward. Malaria cases were mainly imported from Africa countries, and most patients were young adult males. Post-elimination, the proportion of migrant workers decreased, while technical and business personnel increased significantly(χ2=17.855, P<0.05). Both time-to-diagnosis and time-to-certification changed markedly after elimination, with township health centers showing improved diagnostic accuracy at initial visits. All confirmed patients were treated at designated municipal medical institutions. Conclusions After malaria elimination in Qingjiangpu District, imported malaria remains a serious concern, with cases reported each year. Therefore, it is necessary to continue strengthening the malaria monitoring system, enhance screening and evaluation of key populations, improve health education, raise diagnostic and treatment capacity across all medical institutions, and consolidate the achievements of malaria elimination.

Objective The aim was to evaluate the effects of efforts directed toward controlling Oncomelania hupensis snails on small environmental modification projects in Anqing City to provide a reference for comprehensively controlling O. hupensis in hilly areas where schistosomiasis is endemic. Methods Field and retrospective surveys were conducted to collect data using questionnaires. The data from 35 questionnaires on small-scale environmental modification projects in Anqing City and the effects of snail obtained from monitoring forms were sorted. The data were descriptively analyzed using basic information on environmental modifications and investment costs. The Wilcoxon signed-rank test was used to compare snail status before and after environmental interventions. The Friedman test was applied to evaluate the changes in snail populations in the years after environmental intervention. Non-parametric independent sample tests were adopted to compare differences in the effects of different environmental interventions, environmental characteristics, and between project and control areas. Results A total of 35 environments altered from 2017 to 2023, including the creation of 29 ditches,1 dry land, and 5 pond-weirs. The main environmental interventions included hardening of ditches and pond-weirs as well as tilling with intercropping. The average investment costs of mortared rubble masonry hardening and tilling with intercropping were the highest(147.10 yuan/m2)and lowest(4.62 yuan/m2), respectively. The pre-intervention snail-infested area was 78 900 m2, which decreased to 18 100 m2, a decrease of 77.06%. After environmental intervention, the density and rate of living snails significantly decreased on frames with snails(P<0.001). In ditch environments, the smoothness of the ditch walls and the presence of snails in the upper reaches did not significantly decrease the living snail density; however, the presence of cracks on the inner side of the ditches significantly impacted the decline in the rate(P<0.001). No significant difference was found in the rates at which the living snail density or snail frame occurrence decreased among different environmental modification method. The effects of brick masonry, concrete, and mortared rubble masonry on snail control were significantly stronger than those of chemical snail control(P<0.01), whereas no significant difference was found in the decline in these two rates between project areas using other modification method and control areas using chemical snail control. Conclusions Schistosomiasis-prevention-oriented environmental interventions effectively eliminate snails from hilly snail-infested areas. Cost-effective modification method should be selected based on the environmental type, and post-modification monitoring and maintenance must be strengthened to further increase snail control effectiveness.

Objective To analyze the epidemiological characteristics of poisonous mushroom-related foodborne disease incidents in Chongqing from 2010 to 2024, and to provide a scientific basis for effective prevention and control of mushroom-related foodborne diseases. Methods Data of poisonous mushroom related foodborne disease incidents reported by the CDCs of 39 districts and counties in Chongqing from 2010 to 2024 were collected, and their epidemic characteristics were analyzed. Results A total of 264 poisonous mushroom poisoning outbreaks were reported in Chongqing from 2010 to 2024, resulting in 1 046 cases of illness, 543 hospitalizations, and 15 deaths, with a mortality rate of 1.43%. The peak of poisoning incidents was from June to October, accounting for 89.02% of the total number of incidents. The top three high-incidence areas were Pengshui Miao Tujia Autonomous County, Dianjiang County, and Nanchuan District. Poisonous mushroom poisoning incidents occurring in household settings, accidental picking and ingestion, and cases involving unidentified mushroom species accounted for 89.02%, 83.33%, and 90.15% of all foodborne disease outbreak incidents, respectively. Conclusion Foodborne disease incidents related to poisonous mushrooms in Chongqing City show an upward trend, and the effectiveness of prevention and control needs to be improved. Based on their seasonal, regional and household clustering characteristics, efforts should be made before the peak season of mushroom poisoning in high-risk areas, targeting key populations with health education, monitoring and early warning, food safety supervision, and medical treatment.

Dysregulated RNA splicing is a well-recognized characteristic of colorectal cancer (CRC); however, its intricacies remain obscure, partly due to challenges in profiling full-length transcript variants at the single-cell level. Here, we employ high-depth long-read scRNA-seq to define the full-length transcriptome of colorectal epithelial cells in 12 CRC patients, revealing extensive isoform diversities and splicing alterations. Cancer cells exhibited increased transcript complexity, with widespread 3'-UTR shortening and reduced intron retention. Distinct splicing regulation patterns were observed between intrinsic-consensus molecular subtypes (iCMS), with iCMS3 displaying even higher splicing factor activities and more pronounced 3'-UTR shortening. Furthermore, we revealed substantial shifts in isoform usage that result in alterations of protein sequences from the same gene with distinct carcinogenic effects during tumorigenesis of CRC. Allele-specific expression analysis revealed dominant mutant allele expression in key oncogenes and tumor suppressors. Moreover, mutated PPIG was linked to widespread splicing dysregulation, and functional validation experiments confirmed its critical role in modulating RNA splicing and tumor-associated processes. Our findings highlight the transcriptomic plasticity in CRC and suggest novel candidate targets for splicing-based therapeutic strategies.
Humans ; Colorectal Neoplasms/metabolism* ; RNA Isoforms/metabolism* ; Single-Cell Analysis ; RNA Splicing ; Gene Expression Regulation, Neoplastic ; RNA, Neoplasm/metabolism* ; Transcriptome

Kidney fibrosis is the final common pathway of virtually all chronic kidney disease (CKD). However, despite great progress in recent years, no targeted antifibrotic therapies have been approved. Epidemiologic, clinical, and molecular evidence suggest that aging is a major contributor to the increasing incidence of CKD. Senescent renal tubular cells, fibroblasts, endothelial cells, and podocytes have been detected in the kidneys of patients with CKD and animal models. Nonetheless, although accumulated evidence supports the essential role of cellular senescence in CKD, the mechanisms that promote cell senescence and how senescent cells contribute to CKD remain largely unknown. In this review, we summarize the features of the cellular senescence of the kidney and discuss the possible functions of senescent cells in the pathogenesis of kidney fibrosis. We also address whether pharmacological approaches targeting senescent cells can be used to retard the the progression of kidney fibrosis.
Humans ; Cellular Senescence/physiology* ; Fibrosis ; Renal Insufficiency, Chronic/pathology* ; Kidney/pathology* ; Animals

OBJECTIVE: To explore and quantify the association of hot night exposure during the sperm development period (0-90 lag days) with semen quality. METHODS: A total of 6,640 male sperm donors from 6 human sperm banks in China during 2014-2020 were recruited in this multicenter study. Two indices (i.e., hot night excess [HNE] and hot night duration [HND]) were used to estimate the heat intensity and duration during nighttime. Linear mixed models were used to examine the association between hot nights and semen quality parameters. RESULTS: The exposure-response relationship revealed that HNE and HND during 0-90 days before semen collection had a significantly inverse association with sperm motility. Specifically, a 1 °C increase in HNE was associated with decreased sperm progressive motility of 0.0090 (95% confidence interval [ CI]: -0.0147, -0.0033) and decreased total motility of 0.0094 (95% CI: -0.0160, -0.0029). HND was significantly associated with reduced sperm progressive motility and total motility of 0.0021 (95% CI: -0.0040, -0.0003) and 0.0023 (95% CI: -0.0043, -0.0002), respectively. Consistent results were observed at different temperature thresholds on hot nights. CONCLUSION Our findings highlight the need to mitigate nocturnal heat exposure during spermatogenesis to maintain optimal semen quality.
Humans ; Male ; Semen Analysis ; Adult ; Sperm Motility ; Hot Temperature/adverse effects* ; China ; Middle Aged ; Spermatozoa/physiology* ; Young Adult