Objectives This study investigates the molecular epidemiology and BKPyV-specific immune responses in a healthy population to elucidate the infection dynamics of this ubiquitous virus. Methods A cross-sectional study was conducted on 2,543 healthy individuals (aged 1 day to 94 years). Urinary BKPyV DNA was quantified by qPCR, and genotypes were determined by VP1 gene sequencing. BKPyV genotype 1-b1 VP1-specific antibodies levels in urine and serum were measured by ELISA. Results Urinary BKPyV DNA positivity exhibited a bimodal age distribution: 10.1-10.4% in children (5-14 years) and 34.1% in individuals over 70. Serum and urine anti-BKPyV-IgA levels showed a similar age distribution. In contrast, IgG and IgM antibody levels showed a different age distribution. ROC analysis indicated that serum and urine anti-BKPyV-IgA demonstrated superior performance in predicting urinary BKPyV DNA positivity compared to anti-BKPyV-IgG and anti-BKPyV-IgM. Genotypes I (subtypes I-b1 and I-c) and IV were identified, with genotype I being the most prevalent. BKPyV-specific antibody levels in urine and serum were positively correlated. Anti-BKPyV-IgG2, -IgG3, and -IgA1 subclasses were predominant in both compartments. Conclusions This study characterizes aspects of the systemic and mucosal humoral immune response to BKPyV infection, offering preliminary insights into immunity in the general population.

Abstract Sepsis is a life-threatening organ dysfunction caused by a dysregulated host immune response to infection, and there is currently a lack of early rapid identification and effective treatment methods. During the pathogenesis of sepsis, immune cells such as monocytes exhibit abnormal activation of aerobic glycolysis. However, the mechanism of glycolysis in immune cells during sepsis remains to be elucidated. Here, we investigated the role of glycolysis-related regulatory genes in the development of sepsis. Through analysis of the GEO database, we found that HK3 is significantly elevated in the peripheral blood of sepsis patients. Receiver operating characteristic (ROC) curve analysis demonstrated that HK3, as a novel metabolic checkpoint, serves as an excellent diagnostic biomarker for sepsis. Immune cell infiltration analysis revealed a significant increase in monocyte infiltration in the peripheral blood of sepsis patients. Single-cell RNA sequencing analysis demonstrated a significant increase in HK3 expression in monocytes from the sepsis group compared to the control group. Using an LPS-induced monocyte sepsis model, we found that HK3 boosts glycolytic activity and lactate accumulation. Mechanistically, this enhances inflammatory cytokine secretion through H3K18 lactylation-dependent activation of IL-6 and TNF-α genes. Notably, targeted HK3 knockdown effectively suppressed this pro-inflammatory cascade, highlighting its critical role in sepsis pathogenesis. Our findings not only establish HK3 as a key metabolic regulator in sepsis but also elucidate its molecular mechanism in driving excessive monocyte-mediated inflammation. Moreover, we identify HK3 as a promising therapeutic target for mitigating hyperinflammatory responses in sepsis.

MicroRNA-21 (miR-21) is a promising non-invasive biomarker for tumor diagnosis, but its short length and low abundance make amplification-free detection challenging. Here, we developed a dual-function DNA probe (dfDNA)-enabled split crRNA-Cas12a (SCas12a) system for miR-21 detection. This design separates crRNA into a constant scaffold fragment (sc-crRNA) and a variable spacer fragment (sp-crRNA), and miR-21 functionally replaces the spacer fragment to activate Cas12a in the presence of dfDNA. The dfDNA acts as both a target-recognition probe and a fluorescent reporter: it hybridizes with miR-21, activates Cas12a, and presents a single-stranded fluorescent overhang that can be cleaved after Cas12a activation. Because target recognition and signal reporting are integrated within a single probe, dfDNA positions the activated Cas12a complex close to the cleavable overhang, improving signal conversion compared with the conventional single-stranded DNA reporter and fully complementary double-stranded DNA probe format. Under optimized conditions, the limit of detection (LOD) with the homogeneous ssDNA reporter-based assay was 347.9 pM for miR-21, whereas the homogeneous dfDNA-based assay reduced the LOD to 19.8 pM. When transferred to a magnetic bead (MB)-assisted format, the dfDNA-based system achieved an LOD of 1.1 pM, supporting that the localized reaction accelerates the trans-cleavage kinetics of Cas12a. The MB-assisted dfDNA assay achieves comparable or even superior sensitivity for miR-21 detection compared with previously reported split crRNA-based fluorescence methods. Further optimization, including target enrichment or signal amplification strategies, is needed before this method can be reliably used for low-abundance miRNAs in clinical serum samples.

Interpretability in object detection provides crucial confidence support for clinical auxiliary diagnosis. However, in tiny bacteria detection, traditional explanation methods often suffer from blurred foreground boundaries and diffuse feature attribution due to the extreme sparsity of target morphological features and severe interference from complex backgrounds. Such limitations hinder the provision of logically coherent morphological evidence. To bridge this gap, we propose a novel eXplainable AI (XAI) framework, SAM-Sode. The framework innovatively transforms initial feature attribution maps into geometry-aware prompts, leveraging the prior knowledge of the foundation model (SAM3) to achieve spatial refinement and morphological reconstruction of the explanatory mappings. Furthermore, we introduce a dual-constraint mechanism based on physical significance and geometric alignment to perform instance-level denoising, generating coherent explanations that better align with human expert intuition. Experimental results on our self-constructed bacteria dataset with complex circuit backgrounds (containing 2,524 images) and other public datasets demonstrate that the proposed method effectively suppresses background redundancy and significantly enhances the decision-making transparency of tiny object detection.

Gastric cancer (GC) remains one of the leading causes of cancer-related mortality worldwide due to its aggressive nature and high metastatic potential. Circular RNAs (circRNAs), characterized by covalently closed loop structures and high molecular stability, have emerged as important regulatory molecules in cancer biology. However, the biological functions and molecular mechanisms of most circRNAs in GC remain largely unexplored. We conducted circRNA microarray profiling on three pairs of primary GC tissues and matched adjacent normal tissues to identify differentially expressed circRNAs. The expression of circ-ANKRD36C was validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in additional clinical specimens. Functional assays were performed to assess the effects of circ-ANKRD36C knockdown on GC cell proliferation, migration, and invasion. Dual-luciferase reporter assays provided evidence of direct interaction between circ-ANKRD36C and miR-15a-5p. Clinicopathological correlations and survival outcomes were analyzed in 84 paired GC and normal tissue samples. Circ-ANKRD36C was significantly upregulated in GC tissues. Knockdown of circ-ANKRD36C suppressed GC cell proliferation, migration, and invasion in vitro. Dual-luciferase assays supported its direct interaction with miR-15a-5p, potentially influencing oncogenic targets such as BMI1 and CCND1. Clinically, high circ-ANKRD36C expression was associated with shorter overall survival (OS). Multivariate Cox regression analysis suggested its independent prognostic value (hazard ratio [HR] = 2.41; 95

Nitrogen-doped carbon dots (NCD) with high fluorescence retention and good stability were successfully fabricated using citric acid and urea via a facile and eco-friendly one-step microwave method, which exhibited superior specificity for detection of nitrofurantoin (NFT). Upon the addition of NFT, the fluorescence intensity of NCD at 450 nm was significantly decreased. Besides, a satisfactory linear relationship between the fluorescence quenching efficiency and concentrations of NFT was obtained. Especially, NCD was qualitatively and quantitatively applied for detection NFT in milk and meat extract samples with a high recovery rate. Consequently, it was suggested that the detection method had potential application in the specific detection of NFT, offering a novel approach for veterinary drug residue detection.

Diagnosing, predicting disease outcome, and identifying effective treatment targets for virus-related cancers are lacking. Protein biomarkers have the potential to bridge the gap between prevention and treatment for these types of cancers. While it has been shown that certain antibodies against EBV proteins could be used to detect nasopharyngeal carcinoma (NPC), antibodies targeting are solely a tiny part of the about 80 proteins expressed by the EBV genome. Furthermore, it remains unclear what role other viruses play in NPC since many diseases are the result of multiple viral infections. For the first time, this study measured both IgA and IgG antibody responses against 646 viral proteins from 23 viruses in patients with NPC and control subjects using nucleic acid programmable protein arrays. Candidate seromarkers were then validated by ELISA using 1665 serum samples from three clinical cohorts. We demonstrated that the levels of five candidate seromarkers (EBV-BLLF3-IgA, EBV-BLRF2-IgA, EBV-BLRF2-IgG, EBV-BDLF1-IgA, EBV-BDLF1-IgG) in NPC patients were significantly elevated than controls. Additional examination revealed that NPC could be successfully diagnosed by combining the clinical biomarker EBNA1-IgA with the five anti-EBV antibodies. The sensitivity of the six-antibody signature at 95% specificity to diagnose NPC was comparable to the current clinically- approved biomarker combination, VCA-IgA, and EBNA1IgA. However, the recombinant antigens of the five antibodies are easier to produce and standardize compared to the native viral VCA proteins. This suggests the potential replacement of the traditional VCA-IgA assay with the 5-antibodies combination to screen and diagnose NPC. Additionally, we investigated the prognostic significance of these seromarkers titers in NPC. We showed that NPC patients with elevated BLLF3-IgA and BDLF1-IgA titers in their serum exhibited significantly poorer disease-free survival, suggesting the potential of these two seromarkers as prognostic indicators of NPC. These findings will help develop serological tests to detect and treat NPC in the future.

Respiratory pathogens pose a huge threat to public health, especially the highly mutant RNA viruses. Therefore, reliable, on-site, rapid diagnosis of such pathogens is an urgent need. Traditional assays such as nucleic acid amplification tests (NAATs) have good sensitivity and specificity, but these assays require complex sample pre-treatment and a long test time. Herein, we present an on-site biosensor for rapid and multiplex detection of RNA pathogens. Samples with viruses are first lysed in a lysis buffer containing carrier RNA to release the target RNAs. Then, the lysate is used for amplification by one-step reverse transcription and single-direction isothermal strand displacement amplification (SDA). The yield single-strand DNAs (ssDNAs) are visually detected by a lateral flow biosensor. With a secondary signal amplification system, as low as 20 copies/μL of virus can be detected in this study. This assay avoids the process of nucleic acid purification, making it equipment-independent and easier to operate, so it is more suitable for on-site molecular diagnostic applications.

Objectives The specific humoral immune response resulting from inactivated vaccination following by BA.5 infection, and predictors of XBB variants re-infection in BA.5 infection-recovered nasopharyngeal carcinoma (BA.5-RNPC) patients, were explored. Methods Serum SARS-CoV-2 specific antibody levels were assessed using enzyme-linked-immunosorbent-assay. Univariate and multivariate binary logistic regression analyses were conducted to identify factors associated with the magnitude of specific humoral immunity and susceptibility to re-infection by XBB variants. Results Our data demonstrates that SARS-CoV-2 specific antibody levels were comparable between BA.5-RNPC patients and BA.5 infection-recovered-non-cancerous (BA.5-RNC) individuals. Specifically, serum levels of anti-ancestral-S1-IgG, anti-ancestral-nucleocapsid-protein (NP)-IgG, anti-BA.5-receptor binding domain (RBD)-IgG and anti-XBB.1.1.6-RBD-IgG were higher in BA.5-RNPC patients compared to those without a prior infection. Compared to BA.5-RNPC patients without vaccination, individuals who received inactivated vaccination exhibited significantly higher levels of anti-ancestral-S1-IgG and anti-XBB.1.16-RBD-IgG. Multivariate logistic regression analysis revealed that inactivated vaccination was the most significant predictor of all tested SARS-CoV-2 specific antibodies response. Subsequent analysis indicated that a low globulin level is an independent risk factor for XBB re-infection in BA.5-RNPC patients. Conclusions The SARS-CoV-2 specific antibodies have been improved in vaccinated BA.5-RNPC patients. However, the baseline immunity status biomarker IgG is an indicators of XBB variant re-infection risk in BA.5-RNPC patients.

Background C-reactive protein (CRP) is an inflammatory biomarker frequently used in clinical practice. However, insufficient evidence-based ordering inevitably results in its overuse or underuse. This study aims to predict its normal and abnormal levels using the deep neural network (DNN) models, helping clinicians order this item more appropriately and intelligently. Methods We considered complete blood count (CBC) parameters as feature vectors and 10 mg/L as a cutoff value for CRP. Several models, including linear support vector classification, logistic regression, decision trees, random forests, and DNN, were developed based on a dataset of 53834 medical records to predict binary output. We externally validated DNN models on independent 20723 samples through discrimination, calibration curve, and decision curve analysis. Results DNN models has the best area under the receiver operating characteristic curves (AUC). Learning curves revealed that models’ AUC, balanced accuracy, and F1 score do not significantly and continuously improve following increasing data volume. In internal validation, the AUC, balanced accuracy, and the F1 score of 10 models were 0.818 (0.95 CI: 0.812-0.824), 0.741 (0.95 CI: 0.736-0.747), and 0.649 (0.95 CI: 0.643-0.656), respectively. These metrics were 0.817 (0.95 CI: 0.816-0.817), 0.741 (0.95 CI: 0.740-0.742), and 0.641 (0.95 CI: 0.640-0.642), respectively, in external validation. AUC and balanced accuracy shown no significant difference (P-values were 0.106 and 0.339). CRP10-C2 model has the lowest Brier score of 0.154, AUC of 0.818, and calibration curve formula of y=1.001x-0.010, which was identified as a target model to deploy in the app. Conclusions DNN models obtained moderate performance, surpassing baseline indices in distinguishing binary CRP levels. They are good generalizations and well-calibrated. The CRP-C2 model can enhance CRP utilization by informing the orders appropriately and can contribute to inflammatory diagnostics in primary health care where CBC is available, but the CRP test is inaccessible.

Förster resonance energy transfer (FRET)-based homogeneous immunoassay obviates tedious washing steps and thus is a promising approach for immunoassays. However, a conventional FRET-based homogeneous immunoassay operating in the visible region is not able to overcome the interference of complex biological samples, thus resulting in insufficient detection sensitivity and poor accuracy. Here, we develop a near-infrared (NIR)-to-NIR FRET platform (Ex = 808 nm, Em = 980 nm) that enables background-free high-throughput homogeneous quantification of various biomarkers in complex biological samples. This NIR-to-NIR FRET platform is portable and easy to operate and is mainly composed of a high-performance NIR-to-NIR FRET pair based on lanthanide-doped nanoparticles (LnNPs) and a custom-made microplate reader for readout of NIR luminescence signals. We demonstrate that this NIR-to-NIR FRET platform is versatile and robust, capable of realizing highly sensitive and accurate detection of various critical biomarkers, including small molecules (morphine and 1,25-dihydroxyvitamin D), proteins (human chorionic gonadotropin), and viral particles (adenovirus) in unprocessed complex biological samples (urine, whole blood, and feces) within 5-10 min. We expect this NIR-to-NIR FRET platform to provide low-cost healthcare for populations living in resource-limited areas and be widely used in many other fields, such as food safety and environmental monitoring.

Background Currently, there is limited understanding of the specific humoral immune response in BA.5-infected hemodialysis patients (BA.5-CHDPs) with previous COVID-19 vaccination. Additionally, the relevant risk factors for reinfection with XBB variants in BA.5-CHDPs have yet to be elucidated. Method A total of 178 BA.5-CHDPs were enrolled in this study among 53 patients who had previous vaccination. To compare hemodialysis patients in both unvaccinated and vaccinated for their immune response to the BA.5 subtype infection, we assessed serum levels of anti-ancestral-S1-IgG, anti-BA.5-receptor binding domain (RBD)-IgG, and anti-XBB.1.16-RBD-IgG using enzyme-linked immunosorbent assay, the neutralizing antibody titer against BA.5 and XBB.1.16 was determined using pseudovirus neutralization assays. Univariate and multivariate binary logistic regression analyses were conducted to identify factors associated with severe infection, the magnitude of specific humoral immunity and susceptibility to XBB variants reinfection. Result Our findings indicate that BA.5-CHDPs with full or booster vaccinations have higher levels of anti-ancestral-S1-IgG than unvaccinated individuals. However, levels of anti-BA.5-RBD-IgG and anti-XBB.1.16-RBD-IgG are much lower. Booster-vaccinated BA.5-CHDPs have significantly higher levels of BA.5 and XBB.1.16 specific antibodies and neutralizing antibodies than unvaccinated patients. Low globulin levels and shorter hemodialysis duration are independent risk factors for XBB reinfection in BA.5-CHDPs. Conclusion Although XBB.1.16 specific neutralizing antibody levels were low in BA.5-CHDPs, these levels cannot predict the risk of reinfection; other potential risk factors need to be investigated in the future.

The teaching content of network courseware is highly targeted and diversified, which is conducive to students' autonomous learning at any time. At the same time, it can expand students' knowledge and improve the depth of professional knowledge, stimulate students' interest in learning of their own major, thus improving the teaching quality, which is a beneficial supplement to traditional classroom teaching. This paper summarizes the experiences of making network-based multimedia interactive teaching network courseware, characteristics of the network itself, and problems that should be paid attention to in the production process, which is conducive to ensuring the quality of network courseware. With the medical textbook Clinical Basic Examination published by the People's Education Press as the network teaching content, some network courseware has been made. In addition to retaining some teaching content modules of the basic textbook, it further adds network functions such as after-school exercises, reference materials, teaching materials, teacher mailbox, latest teaching and research trends of medicine and laboratory, communication area, and teaching bulletin board module.

Purpose: The aim of this study was to investigate the prognostic significance of PD-1 inhibitor therapy in nasopharyngeal carcinoma (NPC) and to develop a nomogram to estimate individual risks. Methods: We retrospectively analyzed 162 NPC patients who were administered the PD-1 inhibitor combined with radiotherapy and chemotherapy at the Sun Yat-Sen University Cancer Center. In total, 108 NPC patients were included in the training cohort and 54 NPC patients were included in the validation cohort. Univariate and multivariate Cox survival analyses were performed to determine the prognostic factors for 1-year and 2-year progression-free survival (PFS). In addition, a nomogram model was constructed to predict the survival probability of PFS. A consistency index (C-index), a decision curve, a clinical impact curve, and a standard curve were used to measure predictive accuracy, the clinical net benefit, and the consistency of prognostic factors. Results: Univariate and multivariate analyses indicated that the metastasis stage, the levels of ALT, the AST/ALT ratio, and the LDH were independent risk factors associated with the prognosis of PD-1 inhibitor therapy. A nomogram based on these four indicators was constructed and the Kaplan–Meier survival analysis showed that patients with a higher total score have a shorter PFS. The C-index of this model was 0.732 in the training cohort and 0.847 in the validation cohort, which are higher than those for the TNM stages (training cohort: 0.617; validation cohort: 0.727; p <0.05). Decision Curve Analysis (DCA), Net Reclassification Improvement (NRI), and Integrated Discrimination Improvement (IDI) showed that our model has better prediction accuracy than TNM staging. Conclusions: Predicting PFS in NPC patients based on liver function-related indicators before PD-1 treatment may help clinicians predict the efficacy of PD-1 treatment in these patients.

Lateral flow assays (LFAs) are promising points-of-care tests, playing a vital role in diseases screening, diagnosis and surveillance. However, development of portable, cheap, and smart LFAs platform for sensitive and accurate quantification of disease biomarkers in complex media is challenging. Here, a cheap handheld device was developed to realize on-site detection of disease biomarkers by Nd3+/Yb3+ co-doped near-infrared (NIR)-to-NIR downconversion nanoparticles (DCNPs) based LFA. Its sensitivity is at least 8-fold higher for detecting NIR light signal from Nd3+/Yb3+ co-doped nanoparticles than conventional expensive InGaAs camera based detection platform. Additionally, we enhance NIR quantum yield of Nd3+/Yb3+ co-doped nanoparticles up to 35.5% via simultaneous high dopant of sensitizer ions Nd3+ and emitter ions Yb3+. Combination of NIR-to-NIR handheld detection device and ultra-bright NIR emitting NaNbF4:Yb60%@NaLuF4 nanoparticle probe allows the detection sensitivity of SARS-CoV-2 ancestral strain and Omicron variants specific neutralizing antibodies LFA up to the level of commercial enzyme linked immunosorbent assay kit. Furthermore, by this robust method, enhanced neutralizing antibodies against SARS-CoV-2 ancestral strain and Omicron variants are observed in healthy participants with Ad5-nCoV booster on top of two doses of inactivated vaccine. This NIR-to-NIR handheld platform provides a promising strategy for on-site evaluating protective humoral immunity after SARS-CoV-2 vaccination or infection.

Fo center dot rster or fluorescence resonance energy transfer (FRET) enables to probe biomolecular interactions, thus playing a vital role in bioassays. However, conventional FRET platforms suffer from limited sensitivity due to the low FRET efficiency and poor anti-interference of existing FRET pairs. Here we report a NIR-II (1000-1700 nm) FRET platform with extremely high FRET efficiency and exceptional anti-interference capability. This NIR-II FRET platform is established based on a pair of lanthanides downshifting nanoparticles (DSNPs) by employing Nd3+ doped DSNPs as an energy donor and Yb3+ doped DSNPs as an energy acceptor. The maximum FRET efficiency of this well-engineered NIR-II FRET platform reaches up to 92.2%, which is much higher than most commonly used ones. Owing to the all-NIR advantage (lambda ex = 808 nm, lambda em = 1064 nm), this highly efficient NIR-II FRET platform exhibits extraordinary anti-interference in whole blood, and thus enabling background-free ho-mogeneous detection of SARS-CoV-2 neutralizing antibodies in clinical whole blood sample with high sensitivity (limit of detection = 0.5 mu g/mL) and specificity. This work opens up new opportunities for realizing highly sensitive detection of various biomarkers in biological samples with severe background interference.

第五代超文本标记语言(H5)作为 目前颇受欢迎的移动网页开发技术,具有免安装插件、动画效果和交互性强、简单易用等优势.它有很大的应用前景,并且易设计、受欢迎,是一个提高工作效率的实用工具.随着5G时代到来,互联网学习方式的普及,教师移动端教学资源的设计与开发成为备受关注的热点.H5技术的飞速发展为开发跨平台互动多媒体教学资源提供便利与支持.依托H5在线制作平台,教师能轻松制作适用的教学课件、设计微场景或编写测试题,提升课堂教学效果,激发学生的学习兴趣,鼓励学生课后的移动学习和个性化学习,并促进教学改革实践.因此,探讨如何将H5技术与移动端《临床检验基础》多媒体教学资源开发紧密融合有现实意义.

Dear Editor, Recently,a number of new Omicron subvariants related to BA.4/5 and BA.2.75 have emerged and shown remarkable antibody evasion capacities,in particular BF.7,BQ.1,BQ.1.1,BA.2.75.2,XBB and XBB.1.5.1 Unsurprisingly,these new subvariants are quickly gaining prevalence worldwide.

Background The pathogenicity and virulence of the Omicron strain have weakened significantly pathogenesis of Omicron variants. Accumulating data indicated accessory proteins play crucial roles in host immune evasion and virus pathogenesis of SARS-CoV-2. Therefore, the impact of simultaneous deletion of accessory protein ORF7a, ORF7b and ORF8 on the clinical characteristics and specific immunity in Omicron breakthrough infected patients (BIPs) need to be verified. Methods Herein, plasma cytokines were identified using a commercial Multi-cytokine detection kit. Enzyme-linked immunosorbent assay and pseudovirus neutralization assays were utilized to determine the titers of SARS-CoV-2 specific binding antibodies and neutralizing antibodies, respectively. In addition, an enzyme-linked immunospot assay was used to quantify SARS-CoV-2 specific T cells and memory B cells. Results A local COVID-19 outbreak was caused by the Omicron BA.2 variant, which featured a deletion of 871 base pairs (∆871 BA.2), resulting in the removal of ORF7a, ORF7b, and ORF8. We found that hospitalized patients with ∆871 BA.2 had significantly shorter hospital stays than those with wild-type (WT) BA.2. Plasma cytokine levels in both ∆871 BA.2 and WT BA.2 patients were within the normal range of reference, and there was no notable difference in the titers of SARS-CoV-2 ancestor or Omicron-specific binding IgG antibodies, neutralizing antibody titers, effector T cells, and memory B cells frequencies between ∆871 BA.2 and WT BA.2 infected adult patients. However, antibody titers in ∆871 BA.2 infected adolescents were higher than in adults. Conclusions The simultaneous deletion of ORF7a, ORF7b, and ORF8 facilitates the rapid clearance of the BA.2 variant, without impacting cytokine levels or affecting SARS-CoV-2 specific humoral and cellular immunity in Omicron-infected individuals.