As a multiparameter, high-throughput single-cell analysis technology, flow cytometry has become a core tool for biomedical research and clinical diagnosis because of its ability to detect the physical properties (such as size and complexity) and biochemical characteristics (such as surface markers and intracellular proteins) of cells. This article reviews its technical development history, application fields and future challenges. At the technical level, flow cytometry has gradually developed from early single-parameter detection to innovative technologies such as multicolor fluorescence, mass spectrometry flow and imaging flow, realizing the simultaneous analysis and dynamic visualization of over 40 parameters of single cells. In clinical applications, flow cytometry is widely used in the fields of immune diseases and blood tumors, significantly improving diagnostic accuracy and treatment monitoring efficiency. In addition, it also plays a key role in cell biology and drug development. Although flow cytometry faces technical challenges such as high cost, complex data processing and multicolor fluorescence interference, future development directions should focus on intelligence (AI-assisted analysis), portability and multiomics integration, which is expected to further promote the development of precision medicine and personalized treatment. This article systematically reviews the technical evolution and application value of flow cytometry, providing a reference for understanding its core position and future potential in modern medicine.

Self-amplifying RNA (saRNA) represents a novel RNA platform. It has the intrinsic capacity for self-replication by retaining the coding region for the viral replicase. It has this property that it can induce high level of antigen expression even in very low doses and acts as an endogenous immune adjuvant, thus reducing cost and increasing persistence of immunity. But the length of saRNA molecules is large, and they are easily degraded, which leads to problems with delivery efficiency and safety. In addition, the replicase activates an overactive innate immunity. Nanotechnology delivery systems such as lipid nanoparticles (LNPs), cationic nanoemulsions (CNEs) and polymer nanocarriers are important for improving saRNA in vivo stability, cellular uptake and endosome escape. This review focuses on the molecular characteristics of saRNA, the various nanodelivery platforms and optimization strategies employed, and the recent progress made in its therapeutic applications. We also explored the main limitations and future development directions of saRNA, providing a reference for the clinical translation of saRNA.

BACKGROUND:Cancer immunotherapy research, immune microenvironment exploration, and biomarker discovery are key application areas of immune checkpoint analysis. PD-1 and CD28 are crucial receptors expressed on the surface of T cells, playing vital roles in regulating T cell activation and immune response. Accurate detection of these immune checkpoints, such as PD-1 and CD28 on lymphocytes, is essential for understanding immune responses, particularly in clinical contexts such as cancer immunotherapy. Flow cytometry offers a precise approach to detect these markers in whole blood samples. METHODS:This study developed and optimized a flow cytometry-based detection method utilizing the CYTEK NL-CLC flow cytometer to quantitatively assess the expression of PD-1 and CD28 on lymphocytes. A detailed protocol was established and validated, focusing on key performance parameters. RESULTS:Our method showed high sensitivity and specificity, providing a powerful tool for immune monitoring and treatment decision-making. Validation results, including precision, dilution linearity, fluorescence stability, reference interval, and accuracy, all met acceptable criteria and have been reviewed and approved for clinical testing. CONCLUSIONS:The CYTEK NL-CLC flow cytometer is positioned as a reliable and effective platform for immune checkpoint analysis in both clinical and research settings, supporting its integration into cancer immunotherapy workflows and personalized medicine strategies.

Abstract Mucin 1 (MUC1), a prototypical transmembrane mucin, is aberrantly overexpressed and hypoglycosylated in a broad spectrum of epithelial malignancies and hematologic cancers, with its expression level closely correlated with tumor proliferation, metastatic invasion, therapeutic resistance, and poor patient prognosis. This review aims to systematically delineate the structural characteristics and biological functions of MUC1, with a particular focus on the regulatory mechanisms and pathogenic roles of its C-terminal subunit (MUC1-C), and to comprehensively summarize the latest advances in MUC1-targeted anti-tumor therapeutic strategies. We performed an integrated and in-depth synthesis of existing peer-reviewed literature regarding the molecular basis of MUC1 in tumorigenesis, its involvement in oncogenic signaling cascades, and the development of therapeutic modalities targeting this molecule. Upon autoproteolysis, MUC1 is cleaved into two non-covalently linked subunits: the extracellular N-terminal subunit (MUC1-N) that mediates mucosal barrier protection, and the transmembrane MUC1-C, which acts as a central signaling hub to modulate multiple core oncogenic pathways including EGFR, PI3K/AKT/mTOR, NF-κB, and STAT3, thereby inhibiting apoptosis, driving malignant proliferation, epigenetic reprogramming, immune evasion, and inflammation-related pathogenesis. To date, multiple MUC1-targeted therapies, including monoclonal antibodies, antibody–drug conjugates, small molecule inhibitors, CAR-T/NK cell therapies, and cancer vaccines, have advanced to clinical trials, while emerging approaches such as RNA interference, oncolytic virus therapy, and combination immunotherapies have yielded promising anti-tumor efficacy in preclinical in vitro and in vivo studies. Collectively, MUC1, especially the MUC1-C subunit, represents a pivotal and promising target for cancer therapy. Both ongoing clinical investigations and novel MUC1-targeted therapeutic modalities exhibit substantial translational potential, though further in-depth mechanistic research and rigorous clinical validation are still required to optimize their therapeutic efficacy and safety.

Cellular senescence is a distinct and definable biological state characterized by irreversible cell cycle arrest, accompanied by the activation of the DNA damage response (DDR), telomere shortening, the senescence-associated secretory phenotype (SASP), and metabolic dysfunction. While senescent cells represent only a small fraction of the total cell population in tissues, they exert a disproportionate and systemic impact on age-related cardiovascular disease (CVD) through paracrine and endocrine mechanisms. This review moves beyond a descriptive list of pathways and instead proposes a unified framework centered on how a small number of senescent cells can reprogram the cardiovascular microenvironment. We focus on the SASP as the central executor of this systemic effect, disseminating local senescence and driving chronic inflammation, fibrosis, and dysfunction across major cardiovascular cell types (cardiomyocytes, endothelial cells, fibroblasts, smooth muscle cells). We integrate key regulatory networks such as mTOR, AMPK, and Sirtuins that modulate the SASP and the senescent state. Furthermore, we discuss the translational promise of senolytics (agents that clear senescent cells) and senomorphics (agents that suppress the SASP) as novel strategies for delaying cardiovascular aging and treating age-related CVD, providing a forward-looking perspective on targeting senescence to promote cardiovascular health. Current research challenges include mechanistic complexity and limitations of animal models and in vitro systems. In the future, it is necessary to combine single-cell sequencing, metabolic intervention, and interdisciplinary technologies to analyze the heterogeneity of cellular aging, and develop early warning and precision treatment strategies based on aging biomarkers, so as to provide new ideas for delaying cardiovascular aging.

BACKGROUND:This study aims to systematically characterize the physiological trajectories of complete blood cell count (CBC) parameters and their derived inflammatory indicators with age, providing high-quality evidence for assessing immunosenescence and the health of the elderly. METHODS:This cross-sectional study initially recruited 9331 participants aged 24-97 years. After applying exclusion criteria, 1630 healthy individuals were included for analysis. Data are presented as median (2.5th-97.5th percentiles). The Kruskal-Wallis test followed by Dunn's test with Bonferroni correction was used to compare CBC parameters and their derived indicators among four age groups within each sex. The Mann-Whitney U test was used to compare differences between males and females within each age group. RESULTS:Red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), and mean corpuscular hemoglobin concentration (MCHC) decreased with age, while mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and red blood cell distribution width (RDW-CV) increased; sex-specific differences persisted in most parameters. Monocyte percentage and absolute count gradually increased with age; other white blood cell differentials showed variable patterns. Among CBC derived indicators: PLR and SII decreased with age; MLR remained low with limited variability in males; NLR exhibited a fluctuating trend across age groups. Platelet (PLT) progressively decreased with age. CONCLUSION:The comprehensive dataset generated in this study has allowed observation of dynamic profiles of CBC parameters and their derived indicators in the healthy elderly population. This provides scientific evidence and theoretical support for aging and health assessment as well as chronic disease management among older adults.

Chimeric antigen receptor-modified T cell therapy, originally employed in hematological malignancies treatment, has made significant strides in addressing solid tumors in recent years. Presently, second-generation CAR-T therapy has reached clinical implementation, while fifth-generation CAR-T therapy is in active development. However, initial clinical trials in solid tumors have shown limited success, primarily due to the immunosuppressive microenvironment of the tumor and the scarcity of tumor-specific antigens. Nevertheless, emerging strategies, including combination therapies and gene editing technologies, exhibit promising potential in enhancing CAR-T cell effectiveness. In this review, we comprehensively examine the current state of research on CAR-T cell therapy for solid tumors, with a particular focus on the various antigenic targets that have been explored for solid tumors. We critically review established and novel targets, providing insights into their therapeutic potential and limitations. Additionally, we highlight the key challenges that currently hinder the success of CAR-T therapy in solid tumors, including the choice of target antigens, physical barriers, antigen escape, immunosuppressive microenvironment, and adverse reactions. Despite the formidable challenges confronting CAR-T cell therapy in solid tumors, ongoing research is forging a path towards its integration into the mainstream cancer treatment paradigm, offering renewed optimism for enhanced outcomes in patients afflicted with solid malignancies.

BACKGROUND:Depression and cognitive decline are both major health problems worldwide. This study aims to investigate the effects of depressive symptoms and its joint function with lifestyles on cognitive function among the Chinese populations. METHODS:This research included two cross-sectional studies from Hubei (n = 7756) and Shanghai (n = 4751), China. The depressive symptoms were surveyed by the 15-item Geriatric Depression Scale (GDS-15), and cognitive function was assessed by the Mini-mental State Examination (MMSE). RESULTS:We observed significant association between depression and decreased MMSE score [β (95%CI) = -0.70 (-0.97, -0.42) and -3.15 (-3.59, -2.70) in Hubei and Shanghai, respectively] and elevated risk of MCI [OR (95%CI) = 1.59 (1.20, 2.08) and 2.75 (2.06, 3.66), respectively] in both populations. Additionally, individuals who reported physically active, kept an optimal sleep duration, and higher healthy lifestyle score (HLS) exhibited a better cognition in both populations. More importantly, the adverse associations between depressive symptoms and cognition were diminished among subjects adopting five types of healthy lifestyles (HLS = 5) [Hubei: β (95%CI) = -0.20 (-0.82, 0.41) and OR (95%CI) =0.82 (0.24, 2.12), P-int = 0.047 and 0.046; Shanghai: β (95%CI) = -0.80 (-1.70, 0.10) and OR (95%CI) =1.85 (0.53, 5.06), P-int < 0.001 and 0.151]. Furthermore, we observed joint effects of non-depressed status with healthy lifestyles on cognition. CONCLUSION:Depressive symptoms are associated with impaired cognitive function and this association could be attenuated by healthy lifestyles.

BACKGROUND:KL1, KL2 CR-hvKP and KL64, KL47 hv-CRKP all exhibit overlapping multidrug resistance and hypervirulence phenotypes, but the differences in epidemiological, phenotypic and genotypic characteristics between them remains unclear. METHODS:In this study, we collected non-repeated hv-CRKP/CR-hvKP isolates in a tertiary hospital in Shanghai, China from January 2019 to December 2022. Furthermore, we selected four typical hypervirulent and carbapenem-resistant Klebsiella pneumoniae, including ST23-KL1/ST86-KL2 CR-hvKP (WYKP3 and WYKP194) and ST11-KL64/ST11-KL47 hv-CRKP (WYKP589 and WYKP188), and tried to clarify and compare their differences in virulence and drug resistance characteristics and plasmid distribution. RESULTS:Our study found that ST23-KL1 and ST86-KL2 CR-hvKP exhibited less plasmid diversity than that of ST11-KL64 and ST11-KL47 hv-CRKP. Compared with ST11-KL64 and ST11-KL47 hv-CRKP, ST23-KL1/ST86-KL2 CR-hvKP harbored significantly fewer antimicrobial resistance genes but more virulence genes, which contributed to the higher virulence of these strains and exhibited resistance to fewer antibiotics. ST11-KL64 hv-CRKP has emerged as the most prevalent hypervirulent and carbapenem-resistant Klebsiella pneumoniae probably due to its clonal transmission within hospitals as well as the transmission of virulence plasmids with the help of conjugative resistance plasmids. CONCLUSIONS:Due to the different evolutionary mechanisms of hypervirulent and carbapenem-resistant Klebsiella pneumoniae with different capsular serotypes, the epidemiological, phenotypic and genotypic characteristics of KL1, KL2 CR-hvKP and KL64, KL47 hv-CRKP are different.

BackgroundCurrent diagnostic methods have limitations in early prediction of dementia.ObjectiveDevelop an early screening tool to identify persons at high risk of dementia for early intervention.MethodsWe examined amyloid-β protein precursor (AβPP) and its fragments in urine from cognitively normal controls (CNs) and patients with mild cognitive impairment (MCI) or Alzheimer's disease (AD)-dementia using western blotting with different antibodies and developed a colloidal gold lateral flow-based qualitative strip.ResultsCompared with CNs, the amounts of various AβPP fragments with molecular weights of approximately 14, 28, 56, and 68 kDa in patients were greater. We therefore used the strip to detect urine Aβ-containing AβPP fragments and evaluated its potential in multiple aged cohorts from 11 cities in Jiangsu Province, China (n = 4418); Sichuan Provincial People's Hospital (Chengdu, China; n = 408); and the Australian Imaging, Biomarker and Lifestyle Study (AIBL; n = 367). Unitizing Aβ-binding phagocytosis-promoting peptides, the strips showed increasing positivity (9.5-16.9%) with ageing in the Jiangsu cohort and good clinical performance in the Chengdu cohort (κ = 0.704). Significant differences between CNs and patients were found in the AIBL cohort with negative Aβ-PET, those with the slope of Aβ-PET change <1 centiloid per year, those under 75 years of age, or those with a body mass index of 25-30.ConclusionsOur data indicate the high potential of urine AβPP fragments as biomarkers for MCI and dementia at an early stage and warrant further longitudinal studies.

Cancer remains a significant global health challenge, with China being particularly affected because of its large population. Regulated cell death (RCD) mechanisms, including autophagy, apoptosis, necroptosis, pyroptosis, and ferroptosis, play complex roles in cancer development and progression. This review explores the dual roles of autophagy and apoptosis in cancer, highlighting their tumor-suppressive and tumor-promoting functions. Autophagy can maintain genomic stability, induce apoptosis, and suppress protumor inflammation, but it may also support tumor cell survival and drug resistance. Apoptosis, while primarily tumor-suppressive, can paradoxically promote cancer progression in certain contexts. Other RCD mechanisms, such as necroptosis, pyroptosis, and ferroptosis, also exhibit dual roles in cancer, influencing tumor growth, metastasis, and immune responses. Understanding these mechanisms is crucial for developing targeted cancer therapies. This review provides insights into the intricate interplay between RCD mechanisms and cancer, emphasizing the need for context-dependent therapeutic strategies.

Objectives:Accurate and reliable measurement of soluble markers, such as sCD25, sCD40L, sCD130, and sTREM-1, is crucial for understanding their roles in immune responses and inflammatory conditions. This study presents the validation of a multiplex flow cytometry assay using the BD FACSLyric flow cytometer, designed to simultaneously quantify these four soluble markers in serum samples. Methods:The assay utilizes a bead-based immunoassay kit that is compatible with BD FACSLyric to enable precise detection of low concentrations of these markers. The methodological approach includes detailed sample preparation, bead conjugation, and assay optimization. Data acquisition was performed on the BD FACSLyric, with a minimum of 10 000 events recorded per sample to ensure robust data collection. Results:The validation results demonstrated that the assay achieved low limits of detection (LOD) for sCD25, sCD40L, sCD130, and sTREM-1. The LOD values were respectively 9.77 pg/ml, 43.95 pg/ml, 219.73 pg/ml, and 12.21 pg/ml, indicating the assay's high sensitivity. Reproducibility was confirmed through intra-assay and inter-assay variability assessments. Studies have shown that the levels of sCD25/sCD40L/sTREM-1 in the body are associated with lung cancer. Therefore, this study also used this detection method to test both lung cancer patients and healthy individuals, finding significant differences in the results, which can be used to assist in clinical diagnosis and treatment. Conclusions:This assay's ability to detect multiple analytes in a single run, combined with its high sensitivity and reproducibility, makes it a valuable asset for both research and clinical applications. Future work may expand this assay to include additional markers or adapt it for other sample types, further enhancing its utility in diverse biomedical contexts.

BackgroundActive screening for fecal colonization by carbapenem-resistant Enterobacterales (CRE-FC) and interventions in intensive care unit (ICU) wards have become important measures to prevent carbapenem-resistant Enterobacterales (CRE) infection. However, limited data are available on the molecular epidemiology and homology analysis of CRE-FC. This study tried to investigate the molecular epidemiology characteristics and homology of CRE-FC in ICU wards to provide evidence for CRE transmission.MethodsFrom March 1, 2022 to February 28, 2023, fecal swabs from 435 ICU patients were analyzed using resistant bacteria chromogenic plates. Duplicate strains from the same patient were excluded. Infection prevention and control (IPC) measures were implemented for patients with positive CRE screening results. Bacterial identification, antimicrobial susceptibility testing, multilocus sequence typing (MLST), capsule serotypes, whole-genome sequencing (WGS), and core-genome MLST (cgMLST) were conducted to analyze the molecular epidemiological features and homology of these strains.ResultsThe prevalence of CRE-FC in ICU wards was 12.6% (55/435). The predominant CRE-FC was Klebsiella pneumoniae (83.6%, 46/55) and Escherichia coli (9.1%, 5/55). Active screening and IPC interventions in 2022 reduced the CRE infection rate decreased from 11.4 to 7.1%. MLST analysis of 46 carbapenem-resistant K. pneumoniae (CRKP-FC) strains showed that ST11 was the dominant sequence type (71.7%, 33/46), followed by ST15 (26.1%, 12/46) and ST290 (2.2%, 1/46). All ST11 and ST15 strains carried blaKPC-2, 10 of the ST15 strains additionally carried both blaKPC-2 and blaOXA-1. Phylogenetic analysis revealed two major clades: ST11 and ST15.ConclusionCRKP-FC was predominant in CRE-FC. Furthermore, phylogenetic analysis suggested that CRKP-FC had clonal spread in ICU wards, with ST11-KL64 as the main clone. Active CRE-FC screening and IPC interventions can effectively reduced CRE infection rates.

Background It has been reported that the cognitive responses to physical activity (PA) in postmenopausal women vary by parity status, and women with higher parity show a significant association between PA and cognitive function. However, the potential pathways mediating the relationship between PA and cognitive function in women with higher parity remain unclear. The objective of this study was to examine this association in Chinese cohort and further investigate the mediating pathways. Methods A total of 2296 postmenopausal women were enrolled from the Baoshan District, from April to December 2020. All participant details were collected by interviewer-administered questionnaires, including personal information, medical history, lifestyle body mass index (BMI), cognitive function, nutritional status, and. In this cross-sectional study, generalized linear regression models and the chain-mediation analysis were used to examine the relationship between PA and cognitive function and the mediating pathways. Results There was a significant relationship between PA and cognitive function in the high-parity group (≥ three births). In the fully adjusted generalized linear regression model, PA was significantly associated with cognitive function [β: 0.795, 95% confidence interval (CI): 0.251–1.340, P < 0.05]. The chain-mediation analysis showed that depression and nutritional status were two significant mediators, contributing 37.96% of the indirect effect of the overall effect. Conclusions Our findings suggest that PA is beneficial for women at a higher risk of cognitive decline, especially those with more than two births, and these benefits are mediated by depression and nutritional status.

Purpose:To establish a stage-specific diagnostic model for chronic obstructive pulmonary disease (COPD) high-risk individuals by characterizing immune dysregulation through integrated cytokine and lymphocyte profiling. Methods:In this cross-sectional study, 116 participants (34 healthy controls, 56 high-risk individuals, and 26 stable COPD patients) underwent comprehensive immunological evaluation. Peripheral blood cytokine levels (IL-6, IL-8, TNF-α) and lymphocyte subsets-including programmed cell death protein 1-positive (PD-1+) CD4+ T cells and effector memory regulatory T cells (emTregs)-were quantified via flow cytometry and multiplex immunoassays. A multivariable logistic regression model was developed to identify predictors of high-risk COPD, incorporating variables selected through hierarchical likelihood ratio testing and variance inflation factor (VIF)-based multicollinearity adjustment. Statistical validation included receiver operating characteristic (ROC) curve analysis, sensitivity-specificity assessment, and effect size calculations (Cohen's f). Results:Threshold-driven immunological alterations were identified in high-risk individuals, marked by a 7.8-fold elevation in PD-1+CD4+ T cells (p < 0.001) and increased IL-6 levels (median difference: 1.064 pg/mL, p < 0.001). Effector memory Tregs exhibited progressive depletion from healthy to stable COPD stages (p < 0.001). The final regression model-incorporating PD-1+CD4+ T cells, age, and emTregs-demonstrated robust diagnostic accuracy (AUC = 0.912; 95% CI: 0.848-0.975), with 80.9% sensitivity and 79.3% specificity. PD-1+CD4+ T cells and age independently predicted high-risk status (adjusted odds ratio = 1.17, 95% CI: 1.05-1.30, p = 0.005; adjusted odds ratio = 1.11, 95% CI: 1.03-1.20, p = 0.007). Conclusion:This study delineates a threshold-triggered immune signature preceding clinical COPD, providing a validated diagnostic framework for early detection. By integrating lymphocyte exhaustion markers and cytokine dynamics, the model bridges a critical gap in identifying subclinical immune dysfunction, enabling targeted interventions prior to irreversible lung damage.

BACKGROUND:Several studies have reported the association between blood neurofilament light chain (NfL) levels and all-cause mortality. However, the generalizability of these findings in general adults remains unclear. The study aimed to examine the association between serum NfL and all-cause mortality in a nationally representative population. METHODS:Longitudinal data were obtained from 2 071 participants aged 20-75 years in the National Health and Nutrition Examination Survey 2013-14 cycle. Serum NfL levels were measured by using a novel, high-throughput acridinium-ester immunoassay. Kaplan-Meier curves, multivariate Cox regression analysis, and restricted cubic spline regression were employed to investigate the association between serum NfL and all-cause mortality. RESULTS:Over a median follow-up of 73 (interquartile range = 12) months, 85 (3.50%) participants died. After adjustment for sociodemographic characteristics, lifestyle variables, comorbidity, body mass index, and estimated glomerular filtration rate, elevated serum NfL levels were still significantly associated with a higher risk of all-cause mortality (hazard ratio = 2.45, 95% confidence interval = 1.89-3.18 for per lnNfL increase) in a linear manner. CONCLUSIONS:Our findings suggest that circulating levels of NfL may serve as a biomarker of mortality risk in a nationally representative population.

Background:Hypervirulent carbapenem-resistant Klebsiella pneumoniae (hv-CRKP) has recently aroused an extremely severe health challenge and public concern. However, the underlying mechanisms of fitness costs that accompany antibiotic resistance acquisition remain largely unexplored. Here, we report a hv-CRKP-associated fatal infection and reveal a reduction in virulence due to the acquisition of aminoglycoside resistance.Methods:The bacterial identification, antimicrobial susceptibility, hypermucoviscosity, virulence factors, MLST and serotypes were profiled.The clonal homology and plasmid acquisition among hv-CRKP strains were detected by XbaI and S1-PFGE. The virulence potential of the strains was evaluated using Galleria mellonella larvae infection model, serum resistance assay, capsular polysaccharide quantification, and biofilm formation assay. Genomic variations were identified using whole-genome sequencing (WGS).Results:Four K. pneumoniae carbapenemase (KPC)-producing CRKP strains were consecutively isolated from an 86-year-old patient with severe pneumonia. Whole-genome sequencing (WGS) showed that all four hv-CRKP strains belonged to the ST11-KL64 clone. PFGE analysis revealed that the four ST11-KL64 hv-CRKP strains could be grouped into the same PFGE type. Under the pressure of antibiotics, the antimicrobial resistance of the strains increased and the virulence potential decreased. Further sequencing, using the Nanopore platform, was performed on three representative isolates (WYKP586, WYKP589, and WYKP594). Genomic analysis showed that the plasmids of these three strains underwent a large number of breaks and recombination events under antibiotic pressure. We found that as aminoglycoside resistance emerged via acquisition of the rmtB gene, the hypermucoviscosity and virulence of the strains decreased because of internal mutations in the rmpA and rmpA2 genes.Conclusion:This study shows that ST11-KL64 hv-CRKP can further evolve to acquire aminoglycoside resistance accompanied by decreased virulence to adapt to antibiotic pressure in the host.

IntroductionThe chronological age (CA) cannot precisely reflect the health status. Our study aimed to establish a model of kidney biological age to evaluate kidney function more elaborately.MethodsThe modeling group was used to establish the model, consisting of 1,303 respondents of the China Health and Retirement Longitudinal Study (CHARLS). The biological age of the kidney (BA) was constructed by principal component analysis (PCA) and Klemera and Doubal’s method (KDM) with the 1,303 health respondents.ResultsPCA was chosen as the best method for our research step by step. The test group was used to apply the model. (a) BA of the kidney can distinguish respondents with from without kidney disease. (b) BA of the kidney was significantly different in various levels of kidney function. The BA of the eGFR <60 group and 60 ≤ eGFR <90 group were older than GFR ≥90 group. (c) The group with younger BA of kidney at baseline had a lower risk of kidney function decreased. (d) The risk of decreased kidney function caused by increasing BA every additional year is higher than CA.DiscussionThe BA of the kidney is a parameter negatively correlated with decreased kidney function and fills the blank of evaluation among people in the middle of heathy and kidney diseases.

It has been shown that Aß is a normal component in urine (Ghiso et al, FEBS Lett, 1997) and may change with disease status (Takata et al, Neurosci Lett, 2008), however, research on amyloid in urine has been neglected for long time. We examined β amyloid proteins in urine from cognitive normal and dementia patients including Alzheimer’s disease (AD) and frontotemporal dementia (FTD) using Western blotting. Several protein fragments were identified including the 12KD and 14KD bands recognized by the anti-APP C-termini antibody (Ab369), 14KD and 28KD bands by W0-2 (Figure 1), and 28KD, 56KD and 68KD bands by Aducanumab (Figure 2) and anti-APP N-termini antibody (22C11). Further analysis with immunoprecipitation and mass spectrometry confirmed the presence of various APP fragments and Aß in urine. One type of peptide was found to have high affinity to Aß and APP fragments using microscale thermophoresis and circular dichroism spectroscopy. With the assistance from AIBL, we further invented a lateral flow immunoassay device with colloidal gold nanoparticles coated with anti-APP/Aß antibodies. In the early clinical trials with aged people and children, 95% of dementia patients, 90% of MCI, 85% of SMC showed positivity while 90% of healthy children were negative. The second quality examination was performed in Oriental (Dongfeng) Hospital (Pudong, Shanghai) in Mar-June 2022. The examination of detection limit, critical threshold, repeatability, tolerance to interferences (creatinine, protein, glucose, nitrites, pH) was performed and all passed. Furthermore, 43 of 49 cases showed positive (87.7%) while only 1 out of 30 controls was positive (3.3%). Since then, the device has been tested in a dozen of large hospitals and pathology laboratories in China, all provided positive feedback. The preliminary results from 175 AIBL participants showed overall 83.8% positivity to clinically diagnosed MCI (n = 37), while for those Aβ PET scan negative MCI (n = 17), positivity was 94.1% (Figure 3). Our results indicates the high accuracy of this device, which has high potential to be used as an early screening tool for MCI and dementia. The longitudinal studies are currently undergoing in China (Xiangya Hospital) and in Australia (AIBL).