Zhi Chen

Mycoplasma pneumoniae (MP) is an important pathogen associated with respiratory tract infections. Macrolide-resistant MP infections (which mainly contain the A2063G mutation in their 23S rRNA) contribute to severe cases, complications, and sequelae development. Therefore, the development of a sensitive, rapid, and accurate method for MP detection and its A2063G mutation is critical for early differential diagnosis and for guiding clinical medication. Herein, we establish a one-step, highly efficient, and specific fluorescence-based detection system for the MP nucleic acid using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a combined with recombinase polymerase amplification (RPA) isothermal technology. A detection limit of 5 copies/μL within 40 min was achieved using this system. When applied to 57 clinical specimens, this method exhibited a sensitivity of 96.3% and a specificity of 100% compared with quantitative real-time polymerase chain reaction (PCR). We also developed a two-step detection system for macrolide-resistant mutations with a sensitivity of 10 copies/μL. The RPA-CRISPR/Cas13a system was further applied to detect 27 clinical specimens, and the results were consistent with the sequencing data. Additionally, the RPA-CRISPR/Cas13a detection system was integrated with lateral flow assay (LFA) technology to develop an LFA-RPA-CRISPR/Cas13a detection system. In conclusion, our RPA-CRISPR/Cas13a detection system enabled rapid, sensitive, and highly efficient detection of MP and its mutation. This new method provides an efficient tool for clinical and point-of-care testing (POCT) to facilitate early treatment and infection control. IMPORTANCE: A rapid, sensitive, and highly efficient nucleic acid detection system for Mycoplasma pneumoniae (MP) was developed in this study by integrating clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a with recombinase polymerase amplification (RPA) technology. A two-step assay based on this technology was then further developed to identify the A2063G mutation in the 23S rRNA gene of MP that was associated with macrolide resistance. The proposed system was then validated using clinical samples, and it demonstrated consistency with conventional diagnostic and sequencing results. Therefore, the RPA-CRISPR/Cas13a system enabled rapid, sensitive, and cost-effective detection of MP and its drug resistance mutation. This method is a robust tool for clinical and point-of-care testing (POCT) applications. These systems will facilitate early treatment and infection control of MP.

Background and Aim: CD8 ⁺ T cell immunosenescence drives hepatocellular carcinoma (HCC) progression and impedes therapeutic efficacy. We hypothesized that combining interleukin‐15 (IL‐15), which rescues senescent CD8 ⁺ T cells peripherally, with thymosin alpha 1 (Tα1), which replenishes the T cell pool via thymic rejuvenation, may synergistically overcome immunosenescence and enhance antitumor immunity in HCC. Methods: An orthotopic HCC model was established in aged C57BL/6 mice (22–26 months), randomly assigned to receive saline, IL‐15, Tα1, or combined therapy. Tumor progression was monitored by bioluminescence imaging, survival analysis, and histopathology. Hepatic CD8 ⁺ T cell phenotype and function were evaluated by multicolor flow cytometry, immunofluorescence, transcriptomic sequencing, and Western blotting. In vitro validation used primary human CD8 ⁺ T cells co‐cultured with Huh7 hepatoma cells. Results: The combination therapy significantly suppressed tumor growth and prolonged survival. It reduced the proportion of senescent CD8 ⁺ T cells while expanding activated effector populations, enhanced proliferative capacity, and upregulated cytotoxic mediators including granzyme B, perforin, and interferon‐gamma. Transcriptomic and protein‐level analyses revealed that the combination attenuated chronically overactivated phosphatidylinositol 3‐kinase/protein kinase B signaling in hepatic CD8 ⁺ T cells. A protein kinase B agonist, SC79, abrogated these therapeutic effects in vitro, confirming pathway suppression as the key mechanism. Conclusions: Combined IL‐15 and Tα1 therapy reverses CD8 ⁺ T cell senescence and enhances antitumor immunity in HCC through suppression of the phosphatidylinositol 3‐kinase/protein kinase B signaling pathway.

Background: Natural killer (NK) cells serve as pivotal effector cells within the innate immune system, playing an indispensable role against infections and tumors. Individuals with diminished NK cell activity or NK cell deficiency are at a higher risk of developing cancers and experiencing severe viral infections. With global demographic shifts toward aging populations, elucidating the mechanisms of immunosenescence becomes increasingly critical for developing targeted therapeutic interventions against age-related disorders. Main body of the This review provides a comprehensive summary of the phenotypic characteristics, functional changes, and mechanisms of NK cells in aging and diseases. By synthesizing current research, it highlights key pathways contributing to NK cell immunosenescence in the elderly and explores potential strategies to preserve or restore their cytotoxic and immunoregulatory functions. Short conclusion: The review provides novel insights into NK cell immunosenescence and proposes innovative approaches to enhance NK cell activity in aging individuals, offering potential therapeutic avenues for mitigating age-related immune decline.

Nuclear factor erythroid 2-related factor 2 (Nrf2) is an intracellular transcription factor that helps protect against oxidative stress in different types

In recent years, global public health security has encountered significant challenges, with infectious diseases accounting for approximately 25% of global mortality annually. The worldwide pandemic instigated by the novel coronavirus, alongside the persistent threats posed by Ebola, influenza, and multidrug-resistant bacteria, has severely compromised human health, economic development, and social stability. Within this context, the development of rapid and precise pathogen detection technologies has emerged as a critical frontline defense for epidemic prevention and control, serving as a pivotal component in the implementation of the “early detection, early isolation, and early treatment” strategy. The Argonaute (Ago) protein, recognized as a programmable and target-specific activated nuclease, has demonstrated substantial potential in the realm of nucleic acid detection due to its distinctive biological properties, garnering considerable attention. In this study, we delineate the structural characteristics of Ago proteins and elucidate the mechanism underlying their nuclease activity. Furthermore, we review the principles of nucleic acid detection based on Argonaute and provide a comprehensive analysis of recent advancements in related detection systems. Additionally, we compare the advantages of detection based on Argonaute with other detection methodologies. Through a comprehensive analysis, we aim to provide a robust theoretical foundation and an advanced technical reference for the development of new-generation nucleic acid detection platforms with high sensitivity and high specificity.

Hepatocellular carcinoma (HCC) is prevalent globally, and the discovery of new targets is vital for the treatment of HCC. Mitochondrial pyruvate carrier 1 (MPC1) has been found to exhibit reduced expression and promote tumour progression in some cancers, although its role in HCC needs to be explored. Using GEO datasets and Kaplan‒Meier plotter, the clinicopathological features and patient prognosis were analysed by assessing the expression levels of MPC1 in HCC tissues. After MPC1-knockdown and MPC1-overexpressing cell lines were constructed, the effects of modulating MPC1 expression on the malignant phenotype of HCC cells were tested via CCK8, colony formation, and scratch assays and flow cytometry. The effects of MPC1 on HCC cells and mitochondria were examined via MitoTracker Red CMXRos staining, cytoskeleton staining and cellular energy metabolism assays. MPC1 expression was found to be reduced in HCC patients and correlated with prognosis and clinicopathological features. It was found that low expression of MPC1 promotes the malignant phenotype of HCC cells and affects the mitochondrial energy metabolism of HCC cells to support the tumour microenvironment, and that MPC1 may act through signal transducer and activator of transcription 3 (STAT3). MPC1 might play a tumor-suppressing role in HCC through its interaction with STAT3, and this discovery could offer novel perspectives for HCC treatment.

Glutathione peroxidase 4 (GPx4; also known as phospholipid hydroperoxide glutathione peroxidase) inhibits cell death, including apoptosis and ferroptosis, by reducing lipid peroxidation. In addition, western blot assays showed that GPx4 protein levels were elevated in hepatocellular carcinoma (HCC) cells following triptolide (TPL) treatment. Therefore, it was hypothesized that HCC cells might develop partial resistance to TPL‑induced cytotoxicity through upregulation of the GPx4 protein. To enhance anti‑proliferative efficacy, the present study co‑treated HCC cells with a combination of TPL and RAS‑selective lethal 3 (RSL3), a well‑characterized GPx4 activity inhibitor. Subsequent experimental data produced from Cell Counting Kit‑8 and flow cytometric analyses demonstrated that co‑administration of TPL and RSL3 promoted HCC cell apoptosis, elevated intracellular reactive oxygen species levels and induced ferroptosis. These collective findings suggested that co‑treatment with TPL and RSL3 may induce both apoptotic and ferroptotic pathways in HCC cells.

PANoptosis has the features of pyroptosis, apoptosis, and necroptosis. Numerous studies have confirmed the diverse roles of various types of cell death in

Objective: Few comparisons have been implemented between different prenatal care utilization indices and their effects on adverse outcomes. This study investigated the appropriateness of Chinese antenatal care (ANC) regulations and compared Chinese and American adequacy of prenatal care utilization (APNCU) scores.Methods: From 2010 to 2022, the medical records of 60,114 pregnant women were collected from the electronic medical record system (EMRS) in Zhoushan, China. ANC utilization was measured using the APNCU score and five times antenatal care (ANC5). Birth weight outcomes, including small for gestational age (SGA) and large for gestational age (LGA), low birth weight (LBW), macrosomia, birth weight, and preterm birth (PTB), were utilized as outcomes. Multinomial, linear, and logistic regression were used to analyze the association of ANC5 and APNCU with outcomes, respectively. Crossover analysis was implemented to compare the interaction between ANC5 and APNCU on the outcomes.Results: Women who received inadequate prenatal care had increased odds for PTB (ANC5: odds ratio (OR) = 1.12, 95% confidence interval (95%CI) = 1.03–1.21; APNCU: OR = 1.18, 95%CI: 1.07–1.29), delivering SGA infants (ANC5: OR = 1.13, 95%CI = 1.07–1.21; APNCU: OR = 1.11, 95%CI = 1.03–1.20). Crossover analysis revealed that inadequate prenatal care in APNCU only was significantly associated with an increased risk of PTB (OR = 1.48, 95%CI: 1.26–1.73).Conclusion: Women with inadequate prenatal care in ANC5 or APNCU were more likely to suffer from adverse birth outcomes, including PTB, birth weight loss, SGA, and LBW. It indicated that adequate prenatal care is necessary for pregnant women. However, there were interactions between ANC5 and APNCU on PTB, with inadequate prenatal care use by APNCU showing the highest risk of PTB. This indicates that APNCU would be a better tool for evaluating prenatal care use.

Background: To investigate the potential utility of quantitative parameters obtained by ¹⁸F-fibroblast activation protein inhibitor positron emission tomography/computed tomography ([¹⁸F]AlF-NOTA-FAPI-04 PET/CT) in the assessment of organ involvement and disease activity in IgG4-related disease (IgG4-RD). Methods: This study enrolled patients who underwent [¹⁸F]AlF-NOTA-FAPI-04 PET/CT scans at the Department of Rheumatology, The First Affiliated Hospital, Zhejiang University School of Medicine from August 2021 to August 2022. The PET/CT images of the included patients were re-evaluated by PET center technicians, and the maximal standardized uptake value (SUV_max), metabolic lesion volume (MLV), and total lesion FAPI (TL-FAPI) were used to evaluate the involved organs and tissues that abnormally accumulated [¹⁸F]AlF-NOTA-FAPI-04. The clinical and laboratory data of patients are also systematically collected and analyzed. Results: Among the patients included in this study, 12 patients met the IgG4-RD classification criteria established by the American College of Rheumatology in 2019. Among them, 8 were males and 4 were females, with an average age of 59.3 ± 11.5 years. 50% of IgG4-RD patients were found with more organ involvement on PET/CT than physical examination, ultrasonography, and computed tomography. IgG4 levels (Rho = 0.594, p = 0.042) and IgG4-RI (Rho = 0.647, p = 0.023) were significantly positively correlated with TL-FAPI. After linear regression analysis, only TL-FAPI showed a predictive value of RI (R² = 0.356, B = 0.008, p = 0.041). Conclusions: [¹⁸F]AlF-NOTA-FAPI-04 PET/CT is a useful tool for identifying asymptomatic organ involvement and assessing disease activity. The TL-FAPI as an indicator was positively correlated with IgG4-RD disease activity.