Dual-RPA assay for rapid detection and differentiation of E.granulosus and E.multilocularis.

Echinococcus granulosus (Eg) and Echinococcus multilocularis (Em) are the two most widely prevalent types of echinococcosis. Several diagnostic methods have been developed for detecting Eg and Em. However, some limitations, such as being time-consuming, needing expensive instruments, or exhibiting low sensitivity, make these methods unsuitable for on-site detection. In this study, a dual-RPA assay was established to detect and differentiate Eg and Em. The primer concentration ratio, reaction time, and reaction temperature of the dual-RPA were optimized. The result showed that the primer concentration ratio of Eg:Em was 400 nM:400 nM, and the best amplification efficiency was obtained by reacting at 38 °C for 20 min. The sensitivity, specificity, and repeatability of the assay were also tested. The assay's detection limit for both Eg and Em was 10 copies/µL. The assay showed reasonable specificity by testing ten parasitic nucleic acids. The assay's intra- and inter-batch coefficients of variation were below 10%, which indicates robust reproducibility of the assay. Finally, to validate the performance of the dual-RPA assay, it was compared with real-time PCR by using 86 clinical nucleic acid samples. The coincidence rate of Eg between dual-RPA and TaqMan real-time PCR was 96.51%, and the coincidence rate of Em between dual-RPA and TaqMan real-time PCR was 98.84%, indicating its potential for accurate clinical diagnosis. Therefore, this study established a rapid and sensitive dual-RPA assay that can rapidly detect and differentiate Eg and Em in one reaction tube and provided a new assay for the detection of echinococcosis in the field.

Correlation of endolymphatic hydrops and perilymphatic enhancement with the clinical features of Ménière's disease.

OBJECTIVES: To use three-dimensional real inversion recovery (3D-real IR) MRI to investigate correlations between endolymphatic hydrops (EH) grades or the degree of perilymphatic enhancement (PE) and clinical features of Ménière's disease (MD), as previous findings have been inconsistent. METHODS: A total of 273 consecutive patients with definite unilateral MD were retrospectively enrolled from September 2020 to October 2021. All patients underwent 3D-real IR and 3D-T2WI 6 h after intravenous gadolinium injection. MD-related symptom duration and vertigo frequency were recorded. EH grades were evaluated, the signal intensity ratio (SIR) was measured, and correlations between clinical features and EH, PE were assessed respectively. RESULTS: The study included 123 males and 150 females, with a mean age of 53.0 years. A longer duration of vertigo was associated with higher cochlear EH grades, whereas the opposite was true for the duration of aural fullness. A longer time since vertigo onset was associated with higher vestibular EH grades; the opposite was true for the duration of individual vertigo attacks. The multiple regression analysis revealed that age, tinnitus duration, and vestibular EH were risk factors for SIR. Furthermore, the low-frequency hearing threshold (HT) was a risk factor for cochlear and vestibular EH, and the SIR. CONCLUSION: The EH grade and SIR (an indicator for the quantitative evaluation of PE) were correlated with clinical features and HT of MD; thus, imaging can be a valuable tool in planning individualised treatment. CLINICAL RELEVANCE STATEMENT: This study revealed that the grade of endolymphatic hydrops and degree of perilymphatic enhancement positively correlates with the length of time since onset of clinical symptoms and hearing thresholds in patients with Ménière's disease, facilitating the tailored treatment. KEY POINTS: • Relationships between 3-dimensional real inversion recovery features and clinical symptoms in Ménière's disease are unknown. • Symptom duration and hearing thresholds correlated with endolymphatic hydrops grades and degree of perilymphatic enhancement. • MRI features correlate with MD severity; thus, imaging is valuable for planning tailored treatment.

Analysis of 14 ß-agonists in pork using an automated wooden tip-based solid-phase microextraction device and ultra-high-performance liquid chromatography-tandem mass spectrometry.

This study introduces a cost-effective, automated ultra-high-performance liquid chromatography-tandem mass spectrometry method for the detection of 14 ß-agonists in pork using a novel solid-phase microextraction probe composed of polyacrylonitrile and molecularly imprinted polymer. Integrated into an automated extraction device, the probe optimizes extraction prior to analysis while reducing expenses and time compared to traditional solid-phase extraction procedures. The method validation followed the Chinese National Standard (GB/T 27404-2008) and examined limits of detection, limits of quantification, matrix effects, linearity, intraday, and interday precision. Average recovery rates ranged from 71.6% to 82.2%, with relative standard deviations less than 15%. Limits of detection and limits of quantification ranged from 0.09 to 0.39 and 0.27 to 0.99 µg/kg, respectively. The new method identified positive samples more accurately than the current National Standard GB/T 31658.22-2022 and demonstrated its potential for routine assessment and regulatory compliance in the detection of ß-agonists in pork.

Value of fractional-order calculus (FROC) model diffusion-weighted imaging combined with simultaneous multi-slice (SMS) acceleration technology for evaluating benign and malignant breast lesions.

BACKGROUND: This study explores the diagnostic value of combining fractional-order calculus (FROC) diffusion-weighted model with simultaneous multi-slice (SMS) acceleration technology in distinguishing benign and malignant breast lesions. METHODS: 178 lesions (73 benign, 105 malignant) underwent magnetic resonance imaging with diffusion-weighted imaging using multiple b-values (14 b-values, highest 3000 s/mm2). Independent samples t-test or Mann-Whitney U test compared image quality scores, FROC model parameters (D,, ), and ADC values between two groups. Multivariate logistic regression analysis identified independent variables and constructed nomograms. Model discrimination ability was assessed with receiver operating characteristic (ROC) curve and calibration chart. Spearman correlation analysis and Bland-Altman plot evaluated parameter correlation and consistency. RESULTS: Malignant lesions exhibited lower D, and ADC values than benign lesions (P < 0.05), with higher values (P < 0.05). In SSEPI-DWI and SMS-SSEPI-DWI sequences, the AUC and diagnostic accuracy of D value are maximal, with D value demonstrating the highest diagnostic sensitivity, while value exhibits the highest specificity. The D and combined model had the highest AUC and accuracy. D and ADC values showed high correlation between sequences, and moderate. Bland-Altman plot demonstrated unbiased parameter values. CONCLUSION: SMS-SSEPI-DWI FROC model provides good image quality and lesion characteristic values within an acceptable time. It shows consistent diagnostic performance compared to SSEPI-DWI, particularly in D and values, and significantly reduces scanning time.

Multi-b-value DWI to evaluate the synergistic antiproliferation and anti-heterogeneity effects of bufalin plus sorafenib in an orthotopic HCC model.

BACKGROUND: Multi-b-value diffusion-weighted imaging (DWI) with different postprocessing models allows for evaluating hepatocellular carcinoma (HCC) proliferation, spatial heterogeneity, and feasibility of treatment strategies. We assessed synergistic effects of bufalin+sorafenib in orthotopic HCC-LM3 xenograft nude mice by using intravoxel incoherent motion (IVIM), diffusion kurtosis imaging (DKI), a stretched exponential model (SEM), and a fractional-order calculus (FROC) model. METHODS: Twenty-four orthotopic HCC-LM3 xenograft mice were divided into bufalin+sorafenib, bufalin, sorafenib treatment groups, and a control group. Multi-b-value DWI was performed using a 3-T scanner after 3 weeks' treatment to obtain true diffusion coefficient Dt, pseudo-diffusion coefficient Dp, perfusion fraction f, mean diffusivity (MD), mean kurtosis (MK), distributed diffusion coefficient (DDC), heterogeneity index α, diffusion coefficient D, fractional order parameter ß, and microstructural quantity µ. Necrotic fraction (NF), standard deviation (SD) of hematoxylin-eosin staining, and microvessel density (MVD) of anti-CD31 staining were evaluated. Correlations of DWI parameters with histopathological results were analyzed, and measurements were compared among four groups. RESULTS: In the final 22 mice, f positively correlated with MVD (r = 0.679, p = 0.001). Significantly good correlations of MK (r = 0.677), α (r = -0.696), and ß (r= -0.639) with SD were observed (all p < 0.010). f, MK, MVD, and SD were much lower, while MD, α, ß, and NF were higher in bufalin plus sorafenib group than control group (all p < 0.050). CONCLUSION: Evaluated by IVIM, DKI, SEM, and FROC, bufalin+sorafenib was found to inhibit tumor proliferation and angiogenesis and reduce spatial heterogeneity in HCC-LM3 models. RELEVANCE STATEMENT: Multi-b-value DWI provides potential metrics for evaluating the efficacy of treatment in HCC. KEY POINTS: • Bufalin plus sorafenib combination may increase the effectiveness of HCC therapy. • Multi-b-value DWI depicted HCC proliferation, angiogenesis, and spatial heterogeneity. • Multi-b-value DWI may be a noninvasive method to assess HCC therapeutic efficacy.

Aeration-driven piezoelectric activation of peroxymonosulfate achieves effective mitigation of antibiotic resistance dissemination.

The antibiotic resistance dissemination in water has become a globally concerned issue, and the wastewater discharge, especially medical wastewater, is considered as one of the most important sources for antibiotic resistance genes (ARGs). However, the effectiveness of current disinfection techniques in the ARGs reduction still remains controversial. In this study, a novel aeration-driven piezoelectric peroxymonosulfate (PMS) activation system using oxygen-vacancy engineered BaTiO3 (BTO) was developed to effectively eliminate antibiotic resistant bacteria (ARB) and ARGs from water. The ARB can be completely inactivated and ∼3.0 logs of ARGs can be removed by the PMS/BTO/aeration system within 1 h, and the spent BTO nanoparticles can be facilely reused after simple rinsing. The aeration can not only provide the driving force for the piezocatalytic process but also more dissolved oxygen in water that played an important role in the generation of free radicals. The radical quenching experiments and electron spin-resonance (ESR) confirmed that all the free radicals, including singlet oxygen (1O2), hydroxyl radical (OH•), sulfate radical (SO4•-) and superoxide radical (•O2-), contributed to the ARGs reduction and 1O2 radicals were identified as the dominant active species. This work provides a high-efficiency and energy saving approach for the mitigation of ARGs from water as the universal use of aeration in water treatment processes and the good reusability of BTO nanoparticles.

MRI analysis of and factors related to knee injuries in amateur marathon runners.

BACKGROUND: Marathons are the most challenging form of running, and amateur athletes may be more prone to injury due to a lack of professional knowledge and instruction in running. PURPOSE: To analyze the MRI manifestations of and factors related to knee injuries in amateur marathon runners. SUBJECTS: Data were collected from a hospital database of 105 qualified amateur marathon athletes (65 males,40 females), between May 2018 and December 2021. FIELD STRENGTH/SEQUENCE: 1.5T MR: sagittal fs-PDWI, sagittal T1WI and sagittal 3D-DESS sequence. ASSESSMENT: The MRI manifestations of knee joint injury were analyzed and evaluated by two radiologists. STATISTICAL TESTS: The inter-observer agreement on MRI readings was analyzed using the kappa coefficient, and binary logistic regression analysis was employed to identify factors associated with knee injuries. RESULTS: The overall prevalence of knee cartilage lesions, meniscus lesions and bone marrow edema among amateur marathon runners was 45.7%, 72.4%, and 49.5% respectively. Our analysis revealed that older age (OR = 1.135, P<0.001), higher BMI (OR = 1.236, P = 0.044), and slower pace (OR = 2.305, P = 0.017) were associated with increased risk of articular cartilage disease. Furthermore, older age (OR = 1.425, P<0.001) was identified as a risk factor for meniscal lesions, while older age (OR = 1.088, P = 0.002) was bone marrow edema. Notably, no significant correlation was observed between knee joint injuries of amateur marathon athletes and gender or the monthly running distance (P>0.05). CONCLUSIONS: The occurrence of knee injuries among amateur marathon athletes was highly prevalent, with the patellofemoral joint cartilage and posterior horn of medial meniscus being frequently affected areas. Moreover, age, BMI, running years and pace were significant risk factors of knee joint injury.

Identification of crucial roles of transcription factor IhfA on high production of free fatty acids in Escherichia coli.

Transcription factor engineering has unique advantages in improving the performance of microbial cell factories due to the global regulation of gene transcription. Omics analyses and reverse engineering enable learning and subsequent incorporation of novel design strategies for further engineering. Here, we identify the role of the global regulator IhfA for overproduction of free fatty acids (FFAs) using CRISPRi-facilitated reverse engineering and cellular physiological characterization. From the differentially expressed genes in the ihfAL- strain, a total of 14 beneficial targets that enhance FFAs production by above 20 % are identified, which involve membrane function, oxidative stress, and others. For membrane-related genes, the engineered strains obtain lower cell surface hydrophobicity and increased average length of membrane lipid tails. For oxidative stress-related genes, the engineered strains present decreased reactive oxygen species (ROS) levels. These gene modulations enhance cellular robustness and save cellular resources, contributing to FFAs production. This study provides novel targets and strategies for engineering microbial cell factories with improved FFAs bioproduction.

Risk Factors for Distant Metastasis in T3 T4 Rectal Cancer.

Background: Distant metastasis is the leading cause of death in patients with rectal cancer. This study aims to comprehensively analyze the risk factors of distant metastasis in T3 T4 rectal cancer using magnetic resonance imaging (MRI), pathological features, and serum indicators. Methods: The clinicopathological data of 146 cases of T3 T4 rectal cancer after radical resection from January 2015 to March 2023 were retrospectively analyzed. Pre- and postoperative follow-up data of all cases were collected to screen for distant metastatic lesions. Univariate and multivariate Logistic regression methods were used to analyze the relationship between MRI features, pathological results, serum test indexes, and distant metastasis. Results: Of the 146 included patients, synchronous or metachronous distance metastasis was confirmed in 43 (29.4%) cases. The patients' baseline data and univariate analysis showed that mrEMVI, maximum tumor diameter, mr T Stage, pathological N stage, number of lymph node metastasis, cancer nodules, preoperative serum CEA, (Carcinoembryonic antigen) and CA199 were associated with distant metastasis. In the multiple logistic regression model, mrEMVI, pathological N stage, number of lymph node metastasis, maximum tumor diameter, and preoperative serum CEA were identified as independent risk factors for distant metastasis: mrEMVI [odds ratio (OR) = 3.06], pathological N stage (OR = 6.52 for N1 vs N0; OR = 63.47 for N2 vs N0), preoperative serum CEA (OR = 0.27), tumor maximum diameter (OR = 1.03), number of lymph nodes metastasis (OR = 0.62). And, the receiver operating characteristic (ROC) curve was plotted and the area under the curve was calculated (area under the curve [AUC) = 0.817, 95% CI = 0.744-0.890, P < .001]. Conclusions: mrEMVI, pathological N stage, number of lymph node metastasis, maximum tumor diameter and preoperative serum CEA are the independent risk factors for distant metastasis in T3 T4 rectal cancer. A comprehensive analysis of the risk factors for distant metastasis in rectal cancer can provide a reliable basis for formulating individualized treatment strategies, follow-up plans, and evaluating prognosis.

Modified combined short and long axis method versus oblique axis method in adult patients undergoing right internal jugular vein cannulation: A randomized controlled non-inferiority study.

BACKGROUND: Modified combined short and long axis method (MCSL) can replace oblique axis in-plane method (OA-IP) for internal jugular vein cannulation (IJVC). This randomized, non-inferiority study estimated the efficacy of MCSL compared with OA-IP in right IJVC. METHODS: Patients (18-75 yr. old) undergoing right IJVC under local anesthesia were randomly assigned to MCSL or OA-IP group. The primary outcome is the event of first needle pass without posterior vessel wall puncture (PVWP). Secondary outcomes included needle attempts, success rate, puncture and cannulation time, needle visualization, probe placement difficulty and complications. RESULTS: Among 190 randomized patients, 187 were involved in the analysis. The first needle pass without PVWP was 85(89.47%) in the MCSL and 81 (85.26%) in the OA-IP (p = 0.382), with a mean rate difference of 4.2% (95% confidence interval: -5.2-13.6), which confirmed the non-inferiority with the margin of -8%. MCSL group exhibited shorter procedure time and lower complications than OA-IP group. No significant differences were discovered between groups in needle attempts, success rate, incidence of probe placement difficulty and needle visualization. CONCLUSIONS: MCSL is non-inferior to OA-IP in first needle pass without PVWP in adults who underwent elective right IJVC and associate with less complications and shorter operating time. CLINICAL TRIAL REGISTRATION: ChiCTR, ChiCTR2100046899.

Magnetic susceptibility and R2*-based texture analysis for evaluating liver fibrosis in chronic liver disease.

PURPOSE: To explore potential feasibility of texture features in magnetic susceptibility and R2* maps for evaluating liver fibrosis. METHODS: Thirty-one patients (median age 46 years; 22 male) with chronic liver disease were prospectively recruited and underwent magnetic resonance imaging (MRI), blood tests, and liver biopsy. Susceptibility and R2* maps were obtained using a 3-dimensional volumetric interpolated breath-hold examination sequence with a 3T MRI scanner. Texture features, including histogram, gray-level co-occurrence matrix (GLCM), gray-level dependence matrix (GLDM), gray-level run length matrix (GLRLM), gray-level size zone matrix (GLSZM), and neighboring gray tone difference matrix (NGTDM) features, were extracted. Texture features and blood test results of non-significant (Ishak-F < 3) and significant fibrosis patients (Ishak-F ≥ 3) were compared, and correlations with Ishak-F stages were analyzed. Areas under the curve (AUCs) were calculated to determine the efficacy for evaluating liver fibrosis. RESULTS: Nine texture features of susceptibility maps and 19 features of R2* maps were significantly different between non-significant and significant fibrosis groups (all P < 0.05). Large dependence high gray-level emphasis (LDHGLE) of GLDM and long run high gray-level emphasis (LRHGLE) of GLRLM in R2* maps showed significantly negative and good correlations with Ishak-F stages (r = -0.616, P < 0.001; r = -0.637, P < 0.001). Busyness (NGTDM) in susceptibility maps, LDHGLE of GLDM and LRHGLE of GLRLM in R2* maps yield the highest AUCs (AUC = 0.786, P = 0.007; AUC = 0.807, P = 0.004; AUC = 0.819, P = 0.003). CONCLUSION: Texture characteristics of susceptibility and R2* maps revealed possible staging values for liver fibrosis. Susceptibility and R2*-based texture analysis may be a useful and noninvasive method for staging liver fibrosis.