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Purpose: Accurate detection and identification of pathogens and their associated resistance mechanisms are essential prerequisites for implementing precision medicine in the management of Carbapenem-resistant Enterobacterales (CRE). Among the various resistance mechanisms, the production of KPC carbapenemase is the most prevalent worldwide. Consequently, this study aims to develop a convenient and precise nucleic acid detection platform specifically for the blaKPC gene. Methods: The initial phase of our research methodology involved developing a CRISPR/Cas12a detection framework, which was achieved by designing highly specific single-guide RNAs (sgRNAs) targeting the blaKPC gene. To enhance the sensitivity of this system, we incorporated three distinct amplification techniques—polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), and recombinase polymerase amplification (RPA)—into the CRISPR/Cas12a framework. Subsequently, we conducted a comparative analysis of the sensitivity and specificity of these three amplification methods when used in combination with the CRISPR/Cas12a system. Additionally, we assessed the clinical applicability of the methodologies by evaluating fluorescence readouts from 80 different clinical isolates. Furthermore, we employed lateral flow assay technology to provide a visual representation of the results, facilitating point-of-care testing. Results: Following a comparative analysis of the sensitivity and specificity of the three methods, we identified the RPA-Cas12a approach as the optimal detection technique. Our findings demonstrated that the limit of detection (LoD) of the RPA-Cas12a platform was 1 aM (~1 copy/μL) for plasmid DNA and 5 × 10³ fg/μL for genomic DNA. Furthermore, both the sensitivity and specificity of the platform achieved 100% upon validation with 80 clinical isolates. Conclusion: These findings suggest that the developed RPA-Cas12a platform represents a promising tool for the cost-effective, convenient, and accurate detection of the blaKPC gene. [ABSTRACT FROM AUTHOR]

Objective To evaluate five types of DIC scoring systems based on sepsis patients, to explore the values of different DIC scoring systems in the occurrence and prognosis of DIC in sepsis patients, and to compare the applicability of different DIC scoring systems for sepsis complicated with DIC. Methods Laboratory indexes and clinical data from sepsis patients who had been hospitalized in Gansu Provincial People's Hospital from December 1, 2019 to December 31, 2021 were retrospectively analyzed within 24 hours. Five types of DIC scoring systems were used to score, and the difference of diagnostic rate and discharge outcome in sepsis patients with different severity was compared. The ROC curves of five DIC scoring systems were established to evaluate the accuracy of DIC in sepsis patients. Results The fatality rate of sepsis increased with the severity of sepsis (P < 0.05). There were statistically significant differences in discharge outcomes between DIC and non-DIC in the five scoring systems (P < 0.05). JMHW, CDSS and part of ISTH were detected in JAAM cases, while ISTH was detected in non-dominant ISTH cases. ISTH, JAAM, JMHW, CDSS, and non-dominant ISTH5 scoring systems were used to diagnose DIC, and absence of full health restoration and death were 3.0, 3.8, 4.2, 3.9, and 3.0 times higher than non-DIC cases, respectively. Conclusion JAAM scoring system has higher diagnostic rate and sensitivity for adult sepsis. CDSS and JMHW scoring systems are more accurate in predicting the prognosis of sepsis patients. [ABSTRACT FROM AUTHOR]

Background and Objectives: This analytical cross-sectional study investigated the prevalence of vitamin D deficiency and its association with metabolic risk factors in the adult population of Gansu Province, China.Methods and Study Design: In total, 11,157 healthy participants (4,740 men and 6,417 women) were enrolled. A questionnaire was used to assess general characteristics and personal habits. We detected 25-hydroxyvitamin D and associated metabolic parameters through electrochemiluminescence immunoassays.Results: The prevalence of severe deficiency (<10 ng/mL), deficiency (10-20 ng/mL), insufficiency (20-30 ng/mL), and sufficiency (>=30 ng/mL) among the participants was 17.3%, 64.6%, 11.8%, and 6.3%, respectively. Vitamin D deficiency was more prevalent in women than in men (82.5% vs 81.1%, p<0.001). The significant predictors of vitamin D deficiency included younger age and female sex (p<0.05), whereas sun exposure, physical activity, and calcium (Ca) supplementation were associated with less vitamin D deficiency (p<0.05). Serum 25(OH)D3 was inversely associated with parathyroid hormone (r=-0.279, p<0.001) and positively associated with serum Ca (r=0.239, p<0.001), serum P (r=0.090, p=0.018), LDL cholesterol (r=0.100, p=0.008), and BMI (r=0.093, p=0.014). No significant association was observed between serum 25(OH)D3 and metabolic disorders.Conclusions: Vitamin D deficiency is highly prevalent among the adult population of Gansu Province, northwest China, especially young physically inactive and overweight women with limited sunlight exposure, whose biomarkers put them at greater risk of osteoporosis and cardiovascular disease. [ABSTRACT FROM AUTHOR]

The development of antibiotic resistance complicates the treatment of infectious diseases and is a global public health threat. However, drug repurposing can address this resistance issue and reduce research and development costs. Niclosamide is a salicylanilide compound approved by the Food and Drug Administration (FDA), and it has been used clinically for treating parasitic infections for many years. Recent studies have shown that niclosamide can inhibit bacterial and fungus activity by affecting the quorum sensing system, biofilm formation, cell membrane potential, and other mechanisms. Here, we discuss recent advances in the antimicrobial applications of niclosamide and its derivatives to provide new perspectives in treating infectious diseases., Competing Interests: The authors report no conflicts of interest in this work., (© 2024 Liu et al.)

Purpose: Accurate detection and identification of pathogens and their associated resistance mechanisms are essential prerequisites for implementing precision medicine in the management of Carbapenem-resistant Enterobacterales (CRE). Among the various resistance mechanisms, the production of KPC carbapenemase is the most prevalent worldwide. Consequently, this study aims to develop a convenient and precise nucleic acid detection platform specifically for the bla KPC gene., Methods: The initial phase of our research methodology involved developing a CRISPR/Cas12a detection framework, which was achieved by designing highly specific single-guide RNAs (sgRNAs) targeting the bla KPC gene. To enhance the sensitivity of this system, we incorporated three distinct amplification techniques-polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), and recombinase polymerase amplification (RPA)-into the CRISPR/Cas12a framework. Subsequently, we conducted a comparative analysis of the sensitivity and specificity of these three amplification methods when used in combination with the CRISPR/Cas12a system. Additionally, we assessed the clinical applicability of the methodologies by evaluating fluorescence readouts from 80 different clinical isolates. Furthermore, we employed lateral flow assay technology to provide a visual representation of the results, facilitating point-of-care testing., Results: Following a comparative analysis of the sensitivity and specificity of the three methods, we identified the RPA-Cas12a approach as the optimal detection technique. Our findings demonstrated that the limit of detection (LoD) of the RPA-Cas12a platform was 1 aM (~1 copy/µL) for plasmid DNA and 5 × 10³ fg/µL for genomic DNA. Furthermore, both the sensitivity and specificity of the platform achieved 100% upon validation with 80 clinical isolates., Conclusion: These findings suggest that the developed RPA-Cas12a platform represents a promising tool for the cost-effective, convenient, and accurate detection of the bla KPC gene., Competing Interests: The authors report no conflicts of interest in this work, including employment, consultancies, stock ownership, honoraria, paid expert testimony, patent applications/registrations, and grants or other funding., (© 2024 Li et al.)

The Bacteriophage Exclusion (BREX) system is a novel antiphage defense system identified in Bacillus cereus in 2015. The purpose of this study was to investigate the presence of the BREX system defenses against antibiotic-resistant plasmids such as bla KPC and bla NDM invasion in Escherichia coli . The BREX system was present in 5.4% (23/424) of E. coli clinical isolates and 6.5% (84/1283) of E. coli strains with completely sequenced genomes in the GenBank database. All 23 BREX-positive E. coli clinical isolates were susceptible to carbapenems, while all five isolates carrying bla KPC and 11 carrying bla NDM were BREX-negative. For E. coli strains in the GenBank database, 37 of 38 strains carrying bla KPC and 109 of 111 strains carrying bla NDM were BREX negative. The recognition site sequence of methyltransferase PglX in a clinical E. coli 3756 was 5'-CANC A TC-3' using PacBio single-molecular real-time sequencing. The transformation efficiency of plasmid psgRNA-ColAori-target with the PglX recognition site was reduced by 100% compared with the plasmid without the recognition site in E. coli DH5α-pHSG398-BREX. The BREX showed lower defense efficacy against plasmid psgRNA-15Aori-target which had the same plasmid backbone but different surrounding sequences of recognition sites with psgRNA-ColAori-target. The conjugation frequency of the KPC-2 plasmid and NDM-5 plasmid in E. coli 3756-ΔBREX was higher than that in E. coli 3756 clinical isolate (1.0 × 10 -6 vs 1.3 × 10 -7 and 5.5 × 10 -7 vs 1.7 × 10 -8 , respectively). This study demonstrated that the type I BREX system defends against antibiotic-resistant plasmids in E. coli ., Competing Interests: The authors declare no conflict of interest.

In eukaryotes, spliceosomes catalyze the splicing of pre-mRNA to mature mRNA. As the core subunit of U2 spliceosome, splicing factor SF3b4 plays not only a crucial role in the splicing process, but also a role in transcription, translation, and cell signal transduction, and participates in the regulation of cell cycle, cell differentiation, and immune deficiency. In recent years, more and more research studies on SF3b4-related diseases, such as Nager syndrome and cancer, have been conducted. It has been found that SF3b4 mutations led to abnormal cell growth and were involved in the development and occurrence of these diseases. In this review, the diseases, mainly congenital diseases and tumors, in which SF3B4 is involved and the pathogenesis of them were summarized, aiming to provide a better understanding of the roles of SF3B4 in the prevention, diagnosis, and treatment of diseases in the future.