Many antimicrobial resistance genes usually located on transferable plasmids are responsible for multiple antimicrobial resistance among multidrug-resistant (MDR) Gram-negative bacteria. The aim of this study is to characterize a carbapenemase-producing Enterobacter hormaechei 1575 isolate from the blood sample in a tertiary hospital in Wuhan, Hubei Province, China. Antimicrobial susceptibility test showed that 1575 was an MDR isolate. The whole genome sequencing (WGS) and comparative genomics were used to deeply analyze the molecular information of the 1575 and to explore the location and structure of antibiotic resistance genes. The three key resistance genes (blaSFO-1, blaNDM-1, and mcr-9) were verified by PCR, and the amplicons were subsequently sequenced. Moreover, the conjugation assay was also performed to determine the transferability of those resistance genes. Plasmid files were determined by the S1 nuclease pulsed-field gel electrophoresis (S1-PFGE). WGS revealed that p1575-1 plasmid was a conjugative plasmid that possessed the rare coexistence of blaSFO-1, blaNDM-1, and mcr-9 genes and complete conjugative systems. And p1575-1 belonged to the plasmid incompatibility group IncHI2 and multilocus sequence typing ST102. Meanwhile, the pMLST type of p1575-1 was IncHI2-ST1. Conjugation assay proved that the MDR p1575-1 plasmid could be transferred to other recipients. S1-PFGE confirmed the location of plasmid with molecular weight of 342,447 bp. All these three resistant genes were flanked by various mobile elements, indicating that the blaSFO-1, blaNDM-1, and mcr-9 could be transferred not only by the p1575-1 plasmid but also by these mobile elements. Taken together, we report for the first time the coexistence of blaSFO-1, blaNDM-1, and mcr-9 on a transferable plasmid in a MDR clinical isolate E. hormaechei, which indicates the possibility of horizontal transfer of antibiotic resistance genes.
Genetic transformation is one of the key steps in the molecular breeding of chrysanthemum, which relies on an optimal regeneration and transformation system. However, the regeneration system of different chrysanthemum cultivars varies, and the regeneration time of most cultivars is long. To screen cultivars with highly efficient regeneration, leaves and shoot tip thin cell layers (tTCL) from eight chrysanthemum cultivars with different flower colors and flower types were cultured on Murashige and Skoog media (MS) supplemented with 1.0–5.0 mg L−1 6-benzylaminopurine (6-BA) and 0.1–1.0 mg L−1 α-naphthaleneacetic (NAA). The results showed that the most efficient regeneration media were MS + 6-BA 1.0 mg L−1 + NAA 0.5 mg L−1 for leaf explants and MS + 6-BA 5.0 mg L−1 + NAA 0.1 mg L−1 for tTCL explants. Subsequently, another 13 chrysanthemum cultivars were screened by using the media, and finally, three cultivars with high regeneration efficiency were obtained from 21 cultivars. Among these, C1 had the highest regeneration efficiency: the regeneration rate of leaf explants reached 80.0% after 42 days of culture, and the regeneration rate of tTCL explants reached 100% after 31 days of culture. Furthermore, we also established the transformation system for C1 as follows: preculturing for one day, infecting with Agrobacterium suspension (OD600 = 0.6) for 10 min, and cultivating in the regeneration medium with 350 mg L−1 carbenicillin and 10 mg L−1 kanamycin, thus ultimately achieving a transformation rate of 4.0%. In this study, a new chrysanthemum cultivar with an efficient regeneration and transformation system was screened, which is beneficial to enrich the flower color of chrysanthemum transgenic plant recipients and to the functional research of flower color or type-related genes.
The spatial transportation of pesticide spray droplets and their deposition and retention on plant leaf surfaces are critical factors contributing to pesticide loss. Adding adjuvants to pesticide solutions to improve their wettability and deposition behavior can enhance the targeted deposition efficiency of pesticides sprayed by unmanned aerial vehicle (UAV) sprayers. In this study, Maifei(MF), a prevalent vegetable oil adjuvant, was selected to analyze its effects on the physicochemical properties of water and 10% difenoconazole water-dispersible granules (D) and the wetting performance of droplets on litchi leaves. The changes in the drift and deposition of the spray solutions with or without MF were tested using a UAV sprayer, DJI T40. The results indicated that the addition of MF to water or D significantly decreased the surface tension (by 58.33% and 23.10%, respectively), wetting time (by 97.81% and 90.95%, respectively), and contact angle (by 40.95% to 70.75% for the adaxial and abaxial surfaces of litchi leaves), achieving the best effects at a 1% MF addition. Moreover, during the drift test, the addition of 1% MF to the solutions significantly reduced the cumulative drift rate (CDR) (by 48.10%). Finally, owing to the weakened spray drift risk and improved wettability of the droplets on litchi leaves with a 1% MF addition, the droplet deposition and penetration in the litchi canopy significantly improved, demonstrating an increased droplet density of 38.17% for the middle layers of the litchi and 15.75% for the lower layers, corresponding to increased coverage by 59.49% and 12.78%, respectively. Hence, MF can improve the interfacial properties of the spray solution on litchi leaves, reduce the drift risk, and promote deposition, thereby facilitating the efficient transfer and deposition of pesticide droplets from UAV sprayers.
The employment of the no-touch technique in harvesting the great saphenous vein (GSV) for coronary artery bypass grafting has been associated with a significant improvement in clinical patency rates. Despite these advantages, such grafts may predispose patients to complications in the lower limbs. This study endeavors to evaluate the incidence of complications in the lower extremities by deploying an enhanced protocol for the no-touch harvesting technique. The historical control group in this study included patients who underwent coronary artery bypass grafting (CABG) with the no-touch technique for GSV harvesting at our institution from August 2018 to April 2020, in compliance with ethical standards. The intervention group consisted of patients who received CABG and were subjected to an optimized no-touch technique for GSV harvesting from May 2020 to June 2022. Technical modifications were applied to reduce lower limb complications, including limited use of electrocautery, minimization of extravascular tissue preservation, relaxation of postoperative elastic compression bandages, and elevation of the lower extremities. These measures aimed to decrease the incidence of postoperative lower limb complications, such as pain, numbness, edema, exudation, and delayed healing. The occurrences of postoperative complications were meticulously documented, compared, and analyzed between the two groups. The adoption of the optimized no-touch technique resulted in a significant decrease in the incidence of postoperative lower extremity incisional complications among patients subjected to off-pump CABG (p < 0.05). The results of this study substantiate that the application of an optimized no-touch technique to harvest the GSV significantly diminishes the incidence of postoperative lower limb complications in patients receiving CABG. These results highlight the importance of adopting and integrating this optimized technique into clinical protocols, emphasizing its critical role in advancing patient care outcomes.
Abstract Tick-borne encephalitis virus (TBEV) is a serious pathogen that poses a significant threat to humans, causing encephalitis that can result in lifelong sequelae. In this study, we focused on the complete proteomes of the five current TBEV subtypes to identify dominant epitopes. Immunoinformatics tools were employed to screen for LBL, HTL, and CTL epitopes. These epitopes were then linked using various linkers and combined with adjuvants and histidine tag. The vaccine underwent a series of physicochemical property analyses, including secondary structure prediction, three-dimensional structure prediction, molecular docking, molecular dynamics simulation, immune simulation, and in silico cloning. The results indicate that the vaccine is highly conserved, strongly immunogenic, stable, non-allergenic, and non-toxic. Molecular docking and molecular dynamics simulation demonstrate that the vaccine can form a stable binding complex with TLR3. Immune simulation analysis shows that the vaccine effectively stimulates both cellular and humoral immune responses, accompanied by an increase in cytokine titers. Furthermore, through codon optimization and in silico cloning, the vaccine can be stably and effectively expressed in the Escherichia coli system. As an effective candidate for TBEV vaccination, the multi-epitope vaccine developed in this study has promising application prospects and provides a new approach for the research, development, and improvement of vaccines targeting TBEV.
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