Aims: Previous studies suggested a significant relationship between four surrogate indexes of insulin resistance and subsequent type 2 diabetes mellitus (T2DM). But the association of longitudinal changes (denoted as -D) in CVAI (Chinese visceral adiposity index), LAP (lipid accumulation product), TyG (triglyceride-glucose), and TG/HDL-C (triglyceride/ high-density lipoprotein cholesterol) indexes with the risk of T2DM remained uncertain. We aimed to compare the changes in those four surrogate indexes for predicting T2DM in middle-aged and elderly Chinese.Methods: We extracted data from the China Health and Retirement Longitudinal Study (CHARLS). Multivariate logistic regression models were used to estimate odds ratio (OR) with 95% confidence interval (CI) of incident T2DM with four surrogate indexes. The restricted cubic spline analysis was used to examine potential non-linear correlation and visualize the dose-response relationship between four indexes and T2DM. The receiver operator characteristic curve was used to compare the performance of the four indexes to predict T2DM.Results: We enrolled 4,596 participants in total, including 504 (10.97%) with T2DM. Analysis results showed that four surrogate indexes were associated with T2DM, and the multivariate-adjusted ORs (95% CIs) of T2DM were 1.08 (1.00–1.16), 1.47 (1.32-1.63), 1.12 (1.00–1.25), and 2.45 (2.12–2.83) for each IQR (interquartile range) increment in CVAI-D, LAP-D, TG/HDLC-D, and TyG-D, respectively. Restricted cubic spline regression showed a non-linear correlation between four surrogate indexes and the risk of T2DM (p for non-linear < 0.001). From the ROC (receiver operating characteristic) curve, TyG-D had the highest AUC (area under curve), and its AUC values were significantly different from other three indexes both in male and female (all P < 0.001).Conclusion: Compared with other indexes, TyG-D was a better predictor in the clinical setting for identifying middle-aged and elderly Chinese with T2DM. Monitoring long-term changes in TyG might help in the early identification of individuals at high risk of T2DM.

Mutations at RNA splicing sites or regulatory elements can alter splicing efficiency or patterns, affecting RNA functionality and tissue-specific expression. Here, we present a protocol to study the impact of mutations near splicing sites on precursor mRNA (pre-mRNA) splicing. We describe steps for constructing plasmids by cloning the target gene into the pEGFP-N1 vector, performing site-directed mutagenesis, and transiently transfecting HEK293 cells. We then detail procedures for conducting mini-gene splicing assays to analyze splicing patterns influenced by mutations. For complete details on the use and execution of this protocol, please refer to Peng et al.¹.

Rheumatoid arthritis (RA) is a common autoimmune and inflammatory disease. Activated macrophages in arthritic joints play a prominent role in the initiation and persistence of RA. Despite great progress in the clinical treatment of RA, poor response and high discontinuation due to systemic toxicity remain unsolved issues, especially the well-known methotrexate (MTX). Therefore, active targeted delivery of therapeutic drugs to pathogenic cells in arthritic joints is essential to increase in situ activity and decrease systemic toxicity. Here, we developed an MTX-loaded macrophage-targeted nano-emulsion (NE) based on the over-expression of folate receptor (FR) on activated macrophages, the inherent high affinity of FR for folate (FA), as well as the property of MTX and phospholipids to form complexes (MTX@PC). Intravenous injection of DID-labeled MTX@PC-FA NEs into adjuvant-induced arthritis (AIA) mice, in vivo images and flow cytometry results revealed that the NEs were highly targeted to inflamed joints and macrophages, respectively. Therapeutic studies suggested that this strategy was conducive to achieve high efficacy and low toxicity of MTX in the treatment of RA. Our research highlights MTX@PC-FA NEs as a potential treatment option for RA targeting the FR-expressed activated macrophages.

Introduction: Antimicrobial Stewardship programs are crucial for reducing overall antibiotic consumption, but in practice, there are often issues with unclear responsibilities and ambiguous tasks. Pharmacists play a critical role in AMS due to their combined management functions and professional expertise.Objective: To investigate the impact of a Pharmacist-Driven Antimicrobial Stewardship program on the consumption of antibiotics in a hospital.Methods: Under the support of a hospital top-level design, we implemented a Pharmacist-Driven Antimicrobial Stewardship program led by pharmacists and involving multiple disciplines. The program focused on revising the antibiotic formulary and optimizing key points of antibiotic management, using the inpatient Antibiotic Use Density as the core control indicator. This was conducted through three phases: program initiation, implementation, monitoring and control. Clinical pharmacists ensured the long-term operational quality of the program. We evaluated the impact of the program on relevant indicators of antimicrobial consumption in inpatient.Results: Compared to the pre-implementation year of 2020, the annual Antibiotic Use Density for inpatients across the hospital decreased by 22.28% in 2024, reaching 36.26 defined daily doses/100 patient days. Additionally, the monthly inpatient Antibiotic Use Density in 2024 was significantly reduced (p < 0.001), along with the antibiotic usage rate (p < 0.05), expenditure on antibiotics per inpatient (p < 0.001), and the proportion of antibiotic expenditure relative to total for inpatients (p < 0.001). The rational use of antimicrobial agents in inpatient wards has been enhanced. Through targeted management, some antibiotics showed trends of increased or decreased usage. The detection rates of Methicillin-Resistant Staphylococcus aureus and Extended-spectrum β-lactamase-producing Escherichia coli did not show a significant decrease.Conclusion: The Pharmacist-Driven Antimicrobial Stewardship program effectively leveraged the managerial roles and professional skills of pharmacists in rational drug use management, resulting in a significant reduction in hospital antibiotic consumption. However, to further validate its effectiveness in reversing bacterial resistance, the program requires longer-term operation and could be considered for regional expansion.

Neurodegeneration with brain iron accumulation (NBIA) is a clinically and genetically heterogeneous disease characterized by increased iron deposition in the basal ganglia and progressive degeneration of the nervous system in adulthood. However, in early childhood, there were no characteristic features to perform early diagnosis. In our study, a female child exhibited global developmental delay, intellectual disability, and febrile seizure without other distinct clinical phenotypes. Through whole exome sequencing (WES), a de novo nonsense mutation (c.726C > G, p. Tyr242Ter) of WDR45 gene was identified in this child. She was finally diagnosed as β‐propeller protein-associated neurodegeneration (BPAN), one of the recently identified subtypes of NBIA. This mutation could act as a premature stop codon (PSC) which rendered the mutated transcripts to be degraded by nonsense-mediated mRNA decay (NMD), leading to decreased levels of PSC-containing mRNAs. Additionally, through mini-gene splicing assays, this mutation could result in an unprecedented novel transcript with the exon 9 of WDR45 excluded by nonsense-associated splicing alteration (NASA). Transcriptome sequencing (RNA-seq) on total RNAs from PBMCs of the trio revealed three types of alternative splicing events in the patient. Further research implied that downregulation of iron transport genes (TFRC, TFR2, SCARA5) might be the underlying mechanism for the iron accumulation in patients with deficient WDR45. This is the first report about NASA happening in WDR45. It implies that nonsense mutations approximal to splicing sites could affect the disease pathogenesis through more than one molecular mechanism and should be taken into consideration when conducting genetic counseling.