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Summary: IscBs, as hypercompact ancestry proteins of Cas9 nuclease, are suitable for in vivo gene editing via single adeno-associated virus (AAV) delivery. Due to the low activity of natural IscBs in eukaryotic cells, recent studies have been focusing on improving OgeuIscB’s gene editing efficiency via protein engineering. However, in vivo gene editing efficacy of IscBs for disease correction remained to be demonstrated. Here, we showed effective gene knockout and base editing in mouse embryos. To further improve IscB activity, we performed systematic engineering of IscB-associated ωRNA and identified a variant, ωRNA∗-v2, with enhanced gene editing efficiency. Furthermore, our study demonstrated the efficacy of an engineered IscB-ωRNA system for robust gene knockout and base editing in vivo. Single AAV delivery of IscB-derived cytosine and adenine base editors achieved disease correction in a mouse model of tyrosinemia. Therefore, our results indicated the great potential of miniature IscBs for developing single-AAV-based gene editing therapeutics.

Abstract Background Scientific research activity in hospitals is important for promoting the development of clinical medicine, and the scientific literacy of medical staff plays an important role in improving the quality and competitiveness of hospital research. To date, no index system applicable to the scientific literacy of medical staff in China has been developed that can effectively evaluate and guide scientific literacy. This study aimed to establish an index system for the scientific literacy of medical staff in China and provide a reference for improving the evaluation of this system. Methods In this study, a preliminary indicator pool for the scientific literacy of medical staff was constructed through the nominal group technique (n = 16) with medical staff. Then, two rounds of Delphi expert consultation surveys (n = 20) were conducted with clinicians, and the indicators were screened, revised and supplemented using the boundary value method and expert opinions. Next, the hierarchical analysis method was utilized to determine the weights of the indicators and ultimately establish a scientific literacy indicator system for medical staff. Results Following expert opinion, the index system for the scientific literacy of medical staff featuring 2 first-level indicators, 9 second-level indicators, and 38 third-level indicators was ultimately established, and the weights of the indicators were calculated. The two first-level indicators were research literacy and research ability, and the second-level indicators were research attitude (0.375), ability to identify problems (0.2038), basic literacy (0.1250), ability to implement projects (0.0843), research output capacity (0.0747), professional capacity (0.0735), data-processing capacity (0.0239), thesis-writing skills (0.0217), and ability to use literature (0.0181). Conclusions This study constructed a comprehensive scientific literacy index system that can assess medical staff's scientific literacy and serve as a reference for evaluating and improving their scientific literacy.

Abstract Transposon-associated ribonucleoprotein TnpB is known to be the ancestry endonuclease of diverse Cas12 effector proteins from type-V CRISPR system. Given its small size (408 aa), it is of interest to examine whether engineered TnpB could be used for efficient mammalian genome editing. Here, we showed that the gene editing activity of native TnpB from Deinococcus radiodurans (ISDra2 TnpB) in mouse embryos was already higher than previously identified small-sized Cas12f1. Further stepwise engineering of noncoding RNA (ωRNA or reRNA) component of TnpB significantly elevated the nuclease activity of TnpB. Notably, an optimized TnpB-ωRNA system could be efficiently delivered in vivo with single adeno-associated virus (AAV) and corrected the disease phenotype in a tyrosinaemia mouse model. Thus, the engineered miniature TnpB system represents a new addition to the current genome editing toolbox, with the unique feature of the smallest effector size that facilitate efficient AAV delivery for editing of cells and tissues.

Abstract Summary Higher levels of apolipoprotein A1 (ApoA1) were associated with higher risk of osteoporosis, which supports the argument that lipid metabolism is involved in bone metabolism. Background Although the current evidence shows that lipid metabolism and osteoporosis are closely related to cardiovascular disease, the association between ApoA1 and osteoporosis is still unknown. Therefore, the purpose of this study was to explore the relationship between ApoA1 and osteoporosis. Methods In this cross-sectional study, we included 7743 participants in the Third National Health and Nutrition Examination Survey. ApoA1 was regarded as an exposure variable and osteoporosis was considered as an outcome variable. Multivariate logistic regression analysis, sensitivity analysis, and receiver operator characteristic (ROC) were used to assess the association of ApoA1 with osteoporosis. Results Participants with higher ApoA1 had higher rates of osteoporosis compared to participants with lower ApoA1 (P

ObjectiveThe occurrence of Brucella-induced abdominal aortic aneurysms is an exceedingly rare phenomenon, yet it stands as one of the most severe complications within this context. The combined utilization of serological testing and imaging diagnostics has been validated as an effective approach for the identification of Brucella-induced abdominal aortic aneurysms. Presently, the predominant therapeutic strategies encompass antibiotic treatment and surgical intervention. Nonetheless, ongoing controversies persist concerning the establishment of diagnostic criteria, the optimal timing and selection of antibiotic regimens, and the nuanced decision between open surgical procedures and endovascular interventions. Through a meticulous analysis of cases originating from our institution as well as a comprehensive review of previously documented instances, we aim to engage in a detailed discourse on the salient diagnostic and therapeutic facets surrounding Brucella-induced abdominal aortic aneurysms.MethodsWe conducted a retrospective summary of three cases involving Brucella-induced abdominal aortic aneurysms treated within our institution. Furthermore, we performed a comprehensive PubMed search, without imposing restrictions on language or publication year, to identify pertinent literature pertaining to Brucella-induced abdominal aortic aneurysms. The selection criteria primarily focused on case reports delineating occurrences of abdominal aortic aneurysms attributed to Brucella infection.ResultsWe present three distinct cases of Brucella-induced abdominal aortic aneurysms managed at our institution, providing comprehensive insights into the employed diagnostic and therapeutic approaches. Additionally, over the past five decades, a total of 24 cases in 23 publications of Brucella-induced abdominal aortic aneurysms have been reported on PubMed. The earliest report dates back to 1976.ConclusionOur analysis suggests that Brucella-induced abdominal aortic aneurysm is characterized by a remarkably low incidence but is associated with a substantial risk of life-threatening complications. The integration of serological and imaging assessments assumes pivotal importance in facilitating prompt diagnosis of this condition. The prompt initiation of targeted antibiotic therapy is recommended, and the selection of appropriate surgical strategies should be guided by considerations including aneurysm dimensions and morphological attributes. The timely identification and intervention carry utmost significance in retarding disease advancement and ameliorating unfavorable clinical outcomes.

An abdominal aortic aneurysm is a frequently encountered clinical condition, which necessitates prompt and effective remediation to avoid rupture. Surgeons must meticulously select an appropriate method of repair and assess the long-term surgical prognosis when dealing with patients with complex abdominal aortic aneurysms. In this case report, a 74-year-old man was hospitalized due to acute abdominal pain. Upon further examination, it was discovered that he was suffering from a complex abdominal aortic aneurysm. The thoracoabdominal aorta CTA showed that the aneurysm involved both renal arteries, the part below the kidney was severely twisted, the neck of the aneurysm was short, and it was accompanied by bilateral common iliac and internal iliac aneurysms, and there were considerable thrombus attached to the vessel wall. In this case, our team used 3D technology to simulate the spatial structure of the aneurysm and comprehensively evaluate the patient's condition. Ultimately, we decided to perform a quadruple fenestration aortic stent implantation and endovascular repair of aortic aneurysm, combined with right IBE and internal iliac artery stent implantation, right internal iliac artery reconstruction, and left internal iliac artery aneurysm embolization on this patient. This is an innovative surgical method. The operation was successful and the patient recovered well after the operation.

Abstract Background Ischemic strokes are primarily caused by intracranial and extracranial atherosclerotic stenosis. Nontraditional lipid parameters broaden traditional lipid profiles, better reflect the metabolism and interaction between different lipid components, and optimize the predictive ability of lipid profiles for atherosclerotic diseases. This research was carried out to investigate the predictive value of nontraditional lipid parameters for intracranial or extracranial atherosclerotic stenosis. Methods The investigation collected data from inpatients who underwent cervical vascular ultrasonography, carotid CTA, cerebral artery CTA or MRA, and brain MRI or CT from December 2014 to December 2021. The nontraditional lipid parameters were calculated by collecting traditional lipid parameters. To evaluate the predictive power of nontraditional lipid parameters, logistic regression and receiver operating characteristic curve (ROC) analyses were performed. Results Based on the inclusion and exclusion criteria, 545 patients were included. According to the imaging results, inpatients were divided into two groups, including no intracranial or extracranial atherosclerotic stenosis (n = 250) and intracranial or extracranial atherosclerotic stenosis (AS, n = 295). Among them, AS was further divided into three subgroups: intracranial atherosclerotic stenosis (ICAS), extracranial atherosclerotic stenosis (ECAS) and combined intracranial and extracranial atherosclerotic stenosis (IECAS). Logistic regression analysis showed that nontraditional lipid parameters, including the atherogenic index of plasma (AIP), remnant cholesterol (RC), nonhigh-density lipoprotein cholesterol (non-HDL-C), lipoprotein combine index (LCI), atherogenic coefficient (AC), Castelli’s index-I (CRI-I) and Castelli’s index-II (CRI-II), were significantly correlated with intracranial or extracranial atherosclerotic stenosis (P

Summary: The protein level of OCT4, a core pluripotency transcription factor, is vital for embryonic stem cell (ESC) maintenance, differentiation, and somatic cell reprogramming. However, how OCT4 protein levels are controlled during reprogramming remains largely unknown. Here, we identify ubiquitin conjugation sites of OCT4 and report that disruption of WWP2-catalyzed OCT4 ubiquitination or ablation of Wwp2 significantly promotes the efficiency of pluripotency induction from mouse embryonic fibroblasts. Mechanistically, disruption of WWP2-mediated OCT4 ubiquitination elevates OCT4 protein stability and H3K4 methylation level during the reprogramming process. Furthermore, we reveal that OCT4 directly activates expression of Ash2l-b, and that ASH2L-B is a major isoform of ASH2L highly expressed in ESCs and required for somatic cell reprogramming. Together, this study emphasizes the importance of ubiquitination manipulation of the reprogramming factor and its interplay with the epigenetic regulator for successful reprogramming, opening a new avenue to improve the efficiency of pluripotency induction. : In this article, Liao and colleagues show that five lysine residues are major ubiquitination sites of OCT4 catalyzed by WWP2, and that disruption of WWP2-catalyzed OCT4 ubiquitination or ablation of Wwp2 significantly promotes the efficiency of pluripotency induction. Mechanistically, disruption of WWP2-mediated OCT4 ubiquitination elevates OCT4 protein stability and H3K4 methylation level during the reprogramming process. Keywords: Oct4, pluripotency induction, protein stability, ASH2L, histone methylation, ubiquitination

Mesoderm development is a finely tuned process initiated by the differentiation of pluripotent epiblast cells. Serine/threonine kinase 40 (STK40) controls the development of several mesoderm-derived cell types, its overexpression induces differentiation of mouse embryonic stem cells (mESCs) toward the extraembryonic endoderm, and Stk40 knockout (KO) results in multiple organ failure and is lethal at the perinatal stage in mice. However, molecular mechanisms underlying the physiological functions of STK40 in mesoderm differentiation remain elusive. Here, we report that Stk40 ablation impairs mesoderm differentiation both in vitro and in vivo. Mechanistically, STK40 interacts with both the E3 ubiquitin ligase mammalian constitutive photomorphogenesis protein 1 (COP1) and the transcriptional regulator proto-oncogene c-Jun (c-JUN), promoting c-JUN protein degradation. Consequently, Stk40 knockout leads to c-JUN protein accumulation, which, in turn, apparently suppresses WNT signaling activity and impairs the mesoderm differentiation process. Overall, this study reveals that STK40, together with COP1, represents a previously unknown regulatory axis that modulates the c-JUN protein level within an appropriate range during mesoderm differentiation from mESCs. Our findings provide critical insights into the molecular mechanisms regulating the c-JUN protein level and may have potential implications for managing cellular disorders arising from c-JUN dysfunction. [ABSTRACT FROM AUTHOR]