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Objective: To establish a dual fluorescence reverse transcription-enzymatic recombinase amplification (RT-ERA) method for the rapid detection of GI and GII noroviruses (NoV) in shellfish using MS2 as the model process control virus. Methods: The target sequences of GI and GII NoV were individually cloned into vectors with a T7 promoter. Then, high-purity RNA was obtained by in vitro transcription. Dual fluorescence RT-ERA was performed using the primers and probes of MS2 phage designed in this study and the primers and probes of GI and GII NoV selected previously. The reaction procedure and system were optimized. The sensitivity of the method was analyzed. Finally, this method was used to detect real samples, and its accuracy and viability were determined by comparing the results with those obtained by the reference method specified in the national standard GB 4789.42-2016. Results: The optimal volumes of primers and probes for the dual fluorescence RT-ERA assay were 4.1 and 1.8 μL, respectively. The method took only about 10 min to perform, and could detect as low as 102 copies of NoV nucleic acid. When the established method was applied to detect 29 real samples, the results were consistent with those from GB 4789.42-2016. Conclusion: The dual fluorescence RT-ERA assay can simultaneously detect MS2, GI and GII NoV with high sensitivity and accuracy, which will lay a foundation for rapid on-site detection of NoV in the future.

To establish a method to identify the authenticity of manuka honey, this study compared different methods of DNA extraction from mānuka honey pollen, and it developed a DNA extraction method for mānuka honey supernatant and a real-time polymerase chain reaction (real-time PCR) method for the detection of plant-derived ingredients in mānuka honey, including plant internal control, mānuka and kānuka. By analyzing the specificity, sensitivity, detection limit and comparing with the method of the New Zealand Ministry for Primary Industries (MPI), the feasibility, accuracy and equivalence of this method were confirmed. The proposed DNA detection method could replace the MPI method, not only shaking off the dependence on kits, but also making up for the deficiency of identifying mānuka honey pollen only. It has an important application value and scientific significance for the identification of mānuka honey.

Purpose: To investigate the role of Mini Nutritional Assessment-Short Form (MNA-SF) in predicting postoperative complications in older patients (≥ 75 years) undergoing lumbar fusion surgery. Patients and Methods: Patients who had undergone posterior lumbar fusion surgery between June 2019 and September 2021 were enrolled. Those with an MNA-SF score of 12 or higher were categorized as the Nourished group, while those with a score less than 12 were placed in the Malnutrition-Risk group. Preoperative, intraoperative, and postoperative variables between groups were compared. Patients were then re-classified based on the presence of major complications, univariate analysis and multivariate logistic regression was used to identify risk factors for major complications. Results: A total of 240 patients were enrolled, with 182 in the Nourished group and 58 in the Malnutrition-Risk group. The Malnutrition-Risk group exhibited a higher incidence of major complications (46.6% vs 23.1%, p = 0.001) and comprehensive complications index (18.42 ± 18.00 vs 12.65 ± 15.87, p = 0.021), Oswestry Disability Index (27.52 ± 23.44 vs 20.45 ± 20.42, p = 0.029) and longer recovery times (12.53 days vs 10.15 days, p =0.033). Length of stay (LOS) were also increased in the Malnutrition-Risk group (19.22 ± 10.67 vs 16.04 ± 7.69, p = 0.014). Multiple regression analysis identified nutritional risk and malnutrition, as assessed by MNA-SF, as independent factors associated with postoperative major complications (OR 2.81, 95% CI 1.42– 5.53, p = 0.003). Conclusion: Preoperative nutritional risk or malnutrition is an independent risk factor for major complications among older patients undergoing posterior lumbar fusion surgery. The MNS-SF emerges as a convenient and effective tool for promptly screening the nutritional status of older patients, prompting subsequent nutritional evaluation or intervention before surgery. [ABSTRACT FROM AUTHOR]

Background: Lumbar disc degeneration (LDD) is a ubiquitous finding in low back pain. Many different etiology factors may explain the LDD process, such as bone morphogenetic proteins (BMPs), DNA methylation, and gut microbiota. Until recently the mechanisms underlying the LDD process have been elusive. Methods: BMP‐related genes were extracted from the GeneCards database. The LDD transcriptome dataset was obtained from the Gene Expression Omnibus. We used linear regression and meta‐analysis to screen and integrate the differentially expressed genes associated with BMPs in LDD. Genome‐wide association studies (GWASs) of LDD were from FinnGen and UKBB. The expression quantitative trait loci (eQTLs) and DNA methylation quantitative trait loci from the blood were identified via the summary data‐based Mendelian randomization (SMR) method, and the possible blood BMP genes and their regulatory elements associated with the risk of LDD were prioritized. Intestinal eQTLs and fecal microbial QTLs (mbQTLs) were integrated, and the potential interactions between BMP gene expression in host intestinal tissue and the gut microbiota were revealed through SMR and colocalization analysis. The GWAS catalog (GCST90246169) was used to validate SMR results. Results: A meta‐analysis of five datasets revealed that 113 BMP genes were differentially expressed between LDD and control tissues. Seven genes were selected as candidate pathogenic genes of LDD via the three‐step SMR method: CREB1, BMP6, PTCH1, GLI1, MEG3, GALNS, and NF1. SMR analysis also revealed five possible gut genes: HFE, MET, MAPK3, NPC1, and GDF5. The correlation between the gut microbiota and BMP gene expression in intestinal tissues was verified by eQTL‐mbQTL colocalization. Conclusion: This multi‐omics study revealed that the BMP genes associated with LDD are regulated by DNA methylation. There are genetic differences between gut gene expression and the gut microbiota. These findings provide evidence for new therapeutic targets in the future. [ABSTRACT FROM AUTHOR]

Introduction: Besides the comorbid risk factors shared by older and younger patients, older individuals may also experience malnutrition, as well as cognitive or functional impairments. The accumulation of frailty and various geriatric syndromes in older individuals results in decreased physiological reserves, which makes the recovery process after spine surgery particularly challenging. Theoretically, combining the presurgery optimisation provided by a multimodal prehabilitation programme with the reduction of surgical stress provided by an enhanced recovery after surgery (ERAS) programme could improve postoperative recovery of older patients., Methods and Analysis: This is a prospective, multicentre, assessor-blinded, randomised controlled study. Patients who are 75 years of age or older and are scheduled for spinal fusion surgery will be enrolled on three academic medical centres. Regular preadmission education and perioperative ERAS care will be given to participants who were randomised to the control group. Participants randomised to the intervention group will receive multimodal prehabilitation combined with ERAS (PREERAS) management. We will include 164 patients with spinal fusion in three hospitals in China. All included patients will be followed for 90 days after surgery or until death. The primary outcome is the Comprehensive Complication Index (CCI), which ranges from 0 to 100, where a score of 100 indicates death due to complications. Secondary outcomes include length of stay and non-home discharge, rates of postoperative complications and unplanned readmission, North American Spine Society satisfaction, and Oswestry Disability Index/Neck disability index. Ninety-day CCI will be compared between groups using linear regression. Other continuous or categorical outcomes will be compared using linear or logistic regression., Ethics and Dissemination: This study has received ethical approval from the Xuanwu Hospital of Capital Medical University Ethics Committees (2024-088-001). The findings will be submitted to a peer-reviewed journal for publication., Trial Registration Number: NCT06140797., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ Group.)