To enable rapid and accurate point-of-care DNA detection, we have developed a single-step, amplification-free nucleic acid detection platform, a DNA substrate-mediated autocatalysis of CRISPR/Cas12a (DSAC). DSAC makes use of the trans-cleavage activity of Cas12a and target template-activated DNA substrate for dual signal amplifications. DSAC employs two distinct DNA substrate types: one that enhances signal amplification and the other that negatively modulates fluorescent signals. The positive inducer utilizes nicked- or loop-based DNA substrates to activate CRISPR/Cas12a, initiating trans-cleavage activity in a positive feedback loop, ultimately amplifying the fluorescent signals. The negative modulator, which involves competitor-based DNA substrates, competes with the probes for trans-cleaving, resulting in a signal decline in the presence of target DNA. These DNA substrate-based DSAC systems were adapted to fluorescence-based and paper-based lateral flow strip detection platforms. Our DSAC system accurately detected African swine fever virus (ASFV) in swine's blood samples at femtomolar sensitivity within 20 min. In contrast to the existing amplification-free CRISPR/Dx platforms, DSAC offers a cost-effective and straightforward detection method, requiring only the addition of a rationally designed DNA oligonucleotide. Notably, a common ASFV sequence-encoded DNA substrate can be directly applied to detect human nucleic acids through a dual crRNA targeting system. Consequently, our single-step DSAC system presents an alternative point-of-care diagnostic tool for the sensitive, accurate, and timely diagnosis of viral infections with potential applicability to human disease detection.

Background: Multiple studies have suggested a possible connection between the gut microbiota and the development of lymphoma, though the exact nature of this relationship remains unclear. This study aimed to explore whether a causal association exists between gut microbiota and lymphoma.Methods: A bidirectional two-sample Mendelian randomization (MR) approach was conducted to investigate potential causal effects between gut microbiota and various lymphoma subtypes. The primary method employed for MR analysis was inverse variance weighted (IVW), supplemented by additional methods including MR-Egger, weighted median, and weighted mode approaches. The Cochrane Q test, MR-PRESSO global test and MR-Egger intercept test were performed to assess pleiotropy and heterogeneity. Furthermore, a reverse MR analysis was performed to explore potential reverse causal effect.Results: The primary MR analysis identified 36 causal relationships between genetic liabilities in gut microbiota and different lymphoma subtypes. Neither the MR-PRESSO test nor the MR-Egger regression detected any pleiotropy, and Cochran’s Q test indicated no significant heterogeneity.Conclusions: Our MR analysis revealed substantial causal associations between gut microbiota and lymphoma, offering new insights into lymphoma prevention and management microbiota.

Hemophilia, an X-linked monogenic disorder, arises from mutations in the F8 or F9 genes, which encode clotting factor VIII (FVIII) or clotting factor IX (FIX), respectively. As a prominent hereditary coagulation disorder, hemophilia is clinically manifested by spontaneous hemorrhagic episodes. Severe cases may progress to complications such as stroke and arthropathy, significantly compromising patients’ quality of life. Hemophilia has a monogenic nature, coupled with quantifiable therapeutic endpoints and predictable treatment outcomes. These characteristics render it an ideal candidate for gene therapy studies. Currently, Food and Drug Administration (FDA)-approved gene therapies utilize recombinant adeno-associated virus (AAV) vectors to deliver functional transgene cassettes to hepatocytes. These therapies offer distinct advantages: a single intravenous administration achieves sustained FVIII and FIX activity levels, providing robust hemostatic control while markedly enhancing patients’ quality of life. However, several challenges remain, including immunogenicity, thrombotic risks, potential gene integration, and prohibitive costs. Future endeavors should prioritize expanding patient eligibility and integrating precision gene-editing technologies to mitigate these limitations. In this review, we provide a comprehensive overview of recent advances and emerging strategies in hemophilia gene therapy, with a particular focus on clinical translation and technological innovation. Ongoing research in this field remains pivotal to overcome existing barriers, enhance treatment accessibility, and ultimately realize curative potential for patients with hemophilia.

Precursor B-cell acute lymphoblastic leukemia (B-ALL) is the most common cancers in children. Failure of induction chemotherapy is a major factor leading to relapse and death in children with B-ALL. Given the importance of altered metabolites in the carcinogenesis of pediatric B-ALL, studying the metabolic profile of children with B-ALL during induction chemotherapy and in different minimal residual disease (MRD) status may contribute to the management of pediatric B-ALL. We collected paired peripheral blood plasma samples from children with B-ALL at pre- and post-induction chemotherapy and analyzed the metabolomic profiling of these samples by ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS). Healthy children were included as controls. We selected metabolites that were depleted in pediatric B-ALL and analyzed the concentrations in pediatric B-ALL samples. In vitro, we study the effects of the selected metabolites on the viability of ALL cell lines and the sensitivity to conventional chemotherapeutic agents in ALL cell lines. Forty-four metabolites were identified with different levels between groups. KEGG pathway enrichment analyses revealed that dysregulated linoleic acid (LA) metabolism and arginine (Arg) biosynthesis were closely associated with pediatric B-ALL. We confirmed that LA and Arg were decreased in pediatric B-ALL samples. The treatment of LA and Arg inhibited the viability of NALM-6 and RS4;11 cells in a dose-dependent manner, respectively. Moreover, Arg increased the sensitivity of B-ALL cells to L-asparaginase and daunorubicin. Arginine increases the sensitivity of B-ALL cells to the conventional chemotherapeutic drugs L-asparaginase and daunorubicin. This may represent a promising therapeutic approach.

Leukemia is driven by metabolic reprogramming, yet the specific causal roles of plasma metabolites in distinct leukemia subtypes remain unclear. This study employed Mendelian randomization (MR) to explore potential causal links between 690 plasma metabolites (and 143 metabolite ratios) and four leukemia subtypes: ALL, AML, CLL, and CML. Genetic variants from genome-wide association studies served as instrumental variables. Multiple MR approaches, including IVW, MR-Egger, and Weighted Median, along with sensitivity analyses, were applied to ensure robust results. Our findings revealed subtype-specific metabolite associations. In ALL, metabolites such as 3-Hydroxyisobutyrate and γ-Glutamylglutamate showed positive associations, while Phosphocholine and Ceramide showed negative associations. AML was positively linked to GlcNAc/GalNAc and negatively to 1-Methylnicotinamide. CLL showed positive associations with Butyrate/Isobutyrate and Androstenediol Monosulfate, and negative ones with Docosatrienoate and α-Tocopherol to Sulfate ratio. CML exhibited negative associations with Cysteine-Glutathione disulfide and Piperine. Our MR study provides a comprehensive evaluation of the metabolomic landscape of leukemia, identifying subtype-specific causal associations involving pathways such as energy metabolism, amino acid metabolism, lipid signaling, and redox homeostasis. These findings offer insights into potential plasma biomarkers and therapeutic targets, revealing distinct metabolic vulnerabilities that warrant further investigation for precision treatment strategies across leukemia subtypes.

Background: Pediatric leukemia (PL) is a common malignant tumor in childhood. Methotrexate (MTX) is a widely used chemotherapeutic agent for the treatment of this disease. This study was based on the FAERS database and explored the adverse reaction signals of MTX in the treatment of PL. Methods: This study retrieved reports of adverse drug events (ADEs) related to MTX in the treatment of PL from the FAERS database (Q1 2018–Q1 2025). The disproportionality approach, including Reporting Odds Ratio (ROR), Proportional Reporting Ratio (PRR), Bayesian Confidence Propagation Neural Network (BCPNN), and Multi-item Gamma Poisson Shrinker (MGPS), was employed to detect ADE signals. Subgroup analyses were conducted by gender and age groups (infants: <1 year; toddlers: 1–3 years; preschoolers: 4–6 years; school-age: 7–10 years; adolescents: 11–17 years), and time-to-onset of ADEs was analyzed using Weibull distribution fitting. Results: A total of 2,907 reports of ADEs associated with MTX treatment for PL were included. Among these reports, males accounted for the highest proportion (58.3%); the main outcome of ADEs was hospitalization (41.9%). At the SOC level, nervous system disorders accounted for the largest number of reports; at the PT level, febrile neutropenia had the highest report count. Among gender-related ADE signals, confusional state showed greater reporting disproportionality in males. In the age-stratified analysis, the leading PTs varied across age groups; 6 signals were detected in all age groups, and reports describing events within 0–30 days accounted for the highest proportion (72.1%). Conclusion: This study identified prominent reporting signals for nervous system disorders, with febrile neutropenia, neurotoxicity, and mucositis among the most frequently reported PTs. Reports involving males and adolescents accounted for larger proportions, and 72.1% of reports with available onset data described events within 30 days. These findings are hypothesis-generating pharmacovigilance signals and do not establish incidence, comparative clinical risk, or causality.