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Abstract Background Non-alcoholic fatty liver disease (NAFLD) is a multifaceted metabolic disorder, whose global prevalence is rapidly increasing. Acetyl CoA carboxylases 1 (ACACA) is the key enzyme that controls the rate of fatty acid synthesis. Hence, it is crucial to investigate the function of ACACA in regulating lipid metabolism during the progress of NAFLD. Methods Firstly, a fatty liver mouse model was established by high-fat diet at 2nd, 12th, and 20th week, respectively. Then, transcriptome analysis was performed on liver samples to investigate the underlying mechanisms and identify the target gene of the occurrence and development of NAFLD. Afterwards, lipid accumulation cell model was induced by palmitic acid and oleic acid (PA ∶ OA molar ratio = 1∶2). Next, we silenced the target gene ACACA using small interfering RNAs (siRNAs) or the CMS-121 inhibitor. Subsequently, experiments were performed comprehensively the effects of inhibiting ACACA on mitochondrial function and lipid metabolism, as well as on AMPK- PPARα- CPT1A pathway. Results This data indicated that the pathways significantly affected by high-fat diet include lipid metabolism and mitochondrial function. Then, we focus on the target gene ACACA. In addition, the in vitro results suggested that inhibiting of ACACA in vitro reduces intracellular lipid accumulation, specifically the content of TG and TC. Furthermore, ACACA ameliorated mitochondrial dysfunction and alleviate oxidative stress, including MMP complete, ATP and ROS production, as well as the expression of mitochondria respiratory chain complex (MRC) and AMPK proteins. Meanwhile, ACACA inhibition enhances lipid metabolism through activation of PPARα/CPT1A, leading to a decrease in intracellular lipid accumulation. Conclusion Targeting ACACA can reduce lipid accumulation by mediating the AMPK- PPARα- CPT1A pathway, which regulates lipid metabolism and alleviates mitochondrial dysfunction.

Abstract Background Traumatic aortic pseudoaneurysms (PSAs) classified as grade III aortic injuries are conventionally repaired as procedural emergencies, generally within 24 h of arrival. These patients typically require adequate resuscitation and treatment of multiple traumatic injuries, which complicate optimal management strategies of aortic PSAs. This study reviews the experience of an Asian single center to evaluate the efficacy and safety of delayed (> 24 h) endovascular repair for PSAs. Methods Twenty-seven patients with blunt aortic injury (BTAI) were brought to our institution between February 2014 and May 2020. Patients with other grades of aortic injuries (grade I, II, or IV) were excluded from the study, and the remaining patients with grade III aortic injuries were placed into the early ( 24 h) groups according to the timing of repair. Medical records and follow-up computed tomography (CT) scans were reviewed to document the outcomes of the procedures. Primary outcomes included mortality and complications. Results During this period, there were 14 patients (13 males and 1 females) with aortic PSAs, and each patient received thoracic endovascular aortic repair (TEVAR). Of these 14 patients, 1 underwent emergent TEVAR, and 13 underwent delayed repair (median 7 days, range, 3–14 days). Over a period of 8 years, the overall survival of our series was 100%. No paraplegia, stroke, ischemia of limb or other serious procedural complications were observed during the duration of follow-up. Conclusion The experience of our center indicates that delayed repair for selected PSAs could be permissible, which enables a repair in more controlled circumstances.

Abstract Background Metacaspases are multifunctional proteins found in plants, fungi and protozoa, and are involved in processes such as insoluble protein aggregate clearance and cell proliferation. Our previous study demonstrated that metacaspase-1 (MCA1) contributes to parasite apoptosis in Toxoplasma gondii. Deletion of MCA1 from T. gondii has no effect on the growth and virulence of the parasites. Three metacaspases were identified in the ToxoDB Toxoplasma Informatics Resource, and the function of metacaspase-2 (MCA2) and metacaspase-3 (MCA3) has not been demonstrated. Methods In this study, we constructed MCA1, MCA2 and MCA1/MCA2 transgenic strains from RHΔku80 (Δku80), including overexpressing strains and knockout strains, to clarify the function of MCA1 and MCA2 of T. gondii. Results MCA1 and MCA2 were distributed in the cytoplasm with punctuated aggregation, and part of the punctuated aggregation of MCA1 and MCA2 was localized on the inner membrane complex of T. gondii. The proliferation of the MCA1/MCA2 double-knockout strain was significantly reduced; however, the two single knockout strains (MCA1 knockout strain and MCA2 knockout strain) exhibited normal growth rates as compared to the parental strain, Δku80. In addition, endodyogeny was impaired in the tachyzoites whose MCA1 and MCA2 were both deleted due to multiple nuclei and abnormal expression of IMC1. We further found that IMC1 of the double-knockout strain was detergent-soluble, indicating that MCA1 and MCA2 are associated with IMC1 maturation. Compared to the parental Δku80 strain, the double-knockout strain was more readily induced from tachyzoites to bradyzoites in vitro. Furthermore, the double-knockout strain was less pathogenic in mice and was able to develop bradyzoites in the brain, which formed cysts and established chronic infection. Conclusion MCA1 and MCA2 are important factors which participate in IMC1 maturation and endodyogeny of T. gondii. The double-knockout strain has slower proliferation and was able to develop bradyzoites both in vitro and in vivo. Graphic abstract

Abstract Background Bacterial ghosts (BGs) are empty bacterial cell envelopes generated by releasing the cellular contents. In this study, a phage infecting Lactobacillus casei ATCC 393 (L. casei 393) was isolated and designated Lcb. We aimed at using L. casei 393 as an antigen delivery system to express phage-derived holin for development of BGs. Results A gene fragment encoding holin of Lcb (hocb) was amplified by polymerase chain reaction (PCR). We used L. casei 393 as an antigen delivery system to construct the recombinant strain pPG-2-hocb/L. casei 393. Then the recombinants were induced to express hocb. The immunoreactive band corresponding to hocb was observed by western-blotting, demonstrating the efficiency and specificity of hocb expression in recombinants. The measurements of optical density at 600 nm (OD600) after induction showed that expression of hocb can be used to convert L. casei cells into BGs. TEM showed that the cytomembrane and cell walls of hocb expressing cells were partially disrupted, accompanied by the loss of cellular contents, whereas control cells did not show any morphological changes. SEM showed that lysis pores were distributed in the middle or at the poles of the cells. To examine where the plasmid DNA was associated, we analyzed the L. casei ghosts loading SYBR Green I labeled pCI-EGFP by confocal microscopy. The result demonstrated that the DNA interacted with the inside rather than with the outside surface of the BGs. To further analyze where the DNA were loaded, we stained BGs with MitoTracker Green FM and the loaded plasmids were detected using EGFP-specific Cy-3-labeled probes. Z-scan sections through the BGs revealed that pCI-EGFP (red) was located within the BGs (green), but not on the outside. Flow cytometry and qPCR showed that the DNA was loaded onto BGs effectively and stably. Conclusions Our study constructed L. casei BGs by a novel method, which may be a promising technology for promoting the further application of DNA vaccine, providing experimental data to aid the development of other Gram-positive BGs.

Abstract Background The purpose of this study was to investigate the influence of different residual meniscus volume on the biomechanics of tibiofemoral joint after discoid lateral meniscus (DLM) surgery by finite element analysis. Methods A knee joint model was established based on CT and MRI imaging data. The DLM model was divided into five regions according to conventional meniscectomy, with volumes of 15%, 15%, 15%, 15%, 15%, and 40% for each region. Additionally, the DLM model was divided into anterior and posterior parts to obtain ten regions. The DLM was resected according to the design scheme, and together with the intact discoid meniscus, a total of 15 models were obtained. Finite element analysis was conducted to assess shear and pressure trends on the knee joint. Results The study observed significant changes in peak shear stress and compressive stress in the lateral meniscus and lateral femur cartilage. As the meniscus volume decreased, there was an increase in these stresses. Specifically, when the meniscus volume reduced to 40%, there was a sharp increase in shear stress (302%) and compressive stress (152%) on the meniscus, as well as shear stress (195%) and compressive stress (157%) on the lateral femur cartilage. Furthermore, the model grouping results showed that preserving a higher frontal volume in the meniscus model provided better biomechanical advantages. Conclusion The use of finite element analysis has demonstrated that preserving more than 55% of the meniscus volume is necessary to prevent a significant increase in joint stress, which can potentially lead to joint degeneration. Additionally, it is crucial to preserve the front volume of the DLM in order to achieve improved knee biomechanical outcomes.