Accelerated discovery and miniaturization of novel single-stranded cytidine deaminases.

Cytidine base editors (CBEs) hold significant potential in genetic disease treatment and in breeding superior traits into animals. However, their large protein sizes limit their delivery by adeno-associated virus (AAV), given its packing capacity of <4.7 kb. To overcome this, we employed a web-based fast generic discovery (WFG) strategy, identifying several small ssDNA deaminases (Sdds) and constructing multiple Sdd-CBE 1.0 versions. SflSdd-CBE 1.0 demonstrated high C-to-T editing efficiency, comparable to AncBE4max, while SviSdd-CBE 1.0 exhibited moderate C-to-T editing efficiency with a narrow editing window (C3 to C5). Utilizing AlphaFold2, we devised a one-step miniaturization strategy, reducing the size of Sdds while preserving their efficiency. Notably, we administered AAV8 expressing PCSK9 targeted sgRNA and SflSdd-CBEs (nSaCas9) 2.0 into mice, leading to gene-editing events (with editing efficiency up to 15%) and reduced serum cholesterol levels, underscoring the potential of Sdds in gene therapy. These findings offer new single-stranded editing tools for the treatment of rare genetic diseases.

Self-Driven, Monopolar Electrohydrodynamic Printing via Dielectric Nanoparticle Layer.

Electrohydrodynamic printing holds both ultrahigh-resolution fabrication capability and unmatched ink-viscosity compatibility yet fails on highly insulating thick/irregular substrates. Herein, we proposed a single-potential driven electrohydrodynamic printing process with submicrometer resolution on arbitrary nonconductive targets, regardless of their geometric shape or sizes, via precoating with an ultrathin dielectric nanoparticle layer. Benefiting from the favorable Maxwell-Wagner polarization, the reversely polarized spot brought about a significant drop (∼57% for ceramics) in the operation voltage as its induced electric field and a negligible residual charge accumulation. Thus, ordered micro/nanostructures with line widths down to 300 nm were directly written at a stage speed as low as 5 mm/s, and silver features with width of ∼2 µm or interval of ∼4 µm were achieved on insulating substrates separately. Flexible sensors and curved heaters were then high-precision printed and demonstrated successfully, presenting this technique with huge potential for fabricating flexible/conformal electronics on arbitrary 3D structures.

Soluble expression of hMYDGF was improved by strain engineering and optimizations of fermentation strategies in Escherichia coli.

Myeloid-derived growth factor (MYDGF) is a cytokine that exhibits a variety of biological functions. This study focused on utilizing BL21(DE3) strain engineering and fermentation strategies to achieve high-level expression of soluble human MYDGF (hMYDGF) in Escherichia coli. Initially, the E. coli expressing strain BL21(DE3) was engineered by deleting the IpxM gene and inserting the GROEL/S and Trigger factor genes. The engineered E. coli strain BL21(TG)/pT-MYDGF accumulated 3557.3 ± 185.6 µg/g and 45.7 ± 6.7 mg/L of soluble hMYDGF in shake flask fermentation, representing a 15.6-fold increase compared to the control strain BL21(DE3)/pT-MYDGF. Furthermore, the yield of hMYDGF was significantly enhanced by optimizing the fermentation conditions. Under optimized conditions, the 5L bioreactor yielded up to 2665.8 ± 164.3 µg/g and 407.6 ± 42.9 mg/L of soluble hMYDGF. The results indicate that the implementation of these optimization strategies could enhance the ratio and yield of soluble proteins expressed by E.coli, thereby meeting the demands of industrial production. This study employed sophisticated strategies to lay a solid foundation for the industrial application of hMYDGF.

Comparative analysis of the femoral neck system (FNS) vs. cannulated cancellous screws (CCS) in the treatment of Middle-aged and elderly patients with femoral neck fractures: clinical outcomes and biomechanical insights.

PURPOSE: This study aimed to compare the clinical outcomes and differences in biomechanical characteristics between the femoral neck system (FNS) and cannulated cancellous screws (CCSs) in the treatment of femoral neck fractures. METHODS: This study retrospectively analysed a cohort of 38 registered cases of femoral neck fractures treated surgically with either the FNS (n = 17) or CCSs (n = 21) between January 2020 and December 2023. Indicators such as fluoroscopy frequency, length of hospital stay, and fracture healing time were compared between the two groups. Functional status was evaluated via the Harris hip score (HHS) and visual analogue scale (VAS), whereas prognosis was assessed based on changes in the neck shaft angle and femoral neck shortening. Additionally, six sets of femoral neck fracture models were developed based on Pauwels angles of 30°, 40°, 50°, 60°, 70°, and 80°. Two experimental groups, FNS and CCS, were established, and a joint reaction force of 1800 N was applied to the proximal femur. The displacement, stress, and stiffness of the components of interest in the different models were tested and compared. RESULTS: The distributions of all the baseline characteristics were similar between the two groups (p > 0.05). The FNS group presented significantly shorter fluoroscopy frequency, length of hospital stay, and fracture healing time (p < 0.05). Harris and VAS scores were higher in the FNS group than in the CCS group (p < 0.05). Postoperative changes in the neck shaft angle and femoral neck shortening were significantly lower in the FNS group than in the CCS group (p < 0.05). The results of the finite element analysis indicated that the maximum stress on the femoral head and varus angle were generally lower in the FNS group than in the CCS group and that the maximum displacement of the femoral head and FNS was generally lower in the FNS group than in the CCS group. However, the superiority of FNS over CCS decreased with increasing Pauwels angle. Additionally, the effectiveness of FNS in limiting displacement of the femoral neck upper wall was not as favourable as that of CCS. CONCLUSIONS: The treatment of femoral neck fractures with FNS is superior and contributes to improved hip joint function. Biomechanical research has confirmed its structural stability and advantages in resisting femoral head varus. However, challenges to its fixation efficacy persist, particularly at higher Pauwels angles.

Efficient degradation of aqueous dichloromethane by an enhanced microbial electrolysis cell: Degradation kinetics, microbial community and metabolic mechanisms.

Dichloromethane (DCM) has been listed as a toxic and harmful water pollutant, and its removal needs attention. Microbial electrolysis cells (MECs) are viewed as a promising alternative for pollutant removal, which can be strengthened from two aspects: microbial inoculation and acclimation. In this study, the MEC for DCM degradation was inoculated with the active sludge enhanced by Methylobacterium rhodesianum H13 (strain H13) and then acclimated in the form of a microbial fuel cell (MFC). Both the introduction of strain H13 and the initiation in MFC form significantly promoted DCM degradation. The degradation kinetics were fitted by the Haldane model, with Vmax, Kh, Ki and vmax values of 103.2 mg/L/hr, 97.8 mg/L, 268.3 mg/L and 44.7 mg/L/hr/cm2, respectively. The cyclic voltammogram implies that DCM redox reactions became easier with the setup of MEC, and the electrochemical impedance spectrogram shows that the acclimated and enriched microbes reduced the charge transfer resistance from the electrode to the electrolyte. In the biofilm, the dominant genera shifted from Geobacter to Hyphomicrobium in acclimation stages. Moreover, Methylobacterium played an increasingly important role. DCM metabolism mainly occurred through the hydrolytic glutathione S-transferase pathway, given that the gene dcmA was identified rather than the dhlA and P450/MO. The exogenous electrons facilitated the reduction of GSSG, directly or indirectly accelerating the GSH-catalyzed dehalogenation. This study provides support for the construction of an efficient and stable MEC for DCM removal in water environment.

Preliminary exploration of the biomechanical properties of three novel cervical porous fusion cages using a finite element study.

BACKGROUND: Porous cages are considered a promising alternative to high-density cages because their interconnectivity favours bony ingrowth and appropriate stiffness tuning reduces stress shielding and the risk of cage subsidence. METHODS: This study proposes three approaches that combine macroscopic topology optimization and micropore design to establish three new types of porous cages by integrating lattices (gyroid, Schwarz, body-centred cubic) with the optimized cage frame. Using these three porous cages along with traditional high-density cages, four ACDF surgical models were developed to compare the mechanical properties of facet articular cartilage, discs, cortical bone, and cages under specific loads. RESULTS: The facet joints in the porous cage groups had lower contact forces than those in the high-density cage group. The intervertebral discs in all models experienced maximum stress at the C5/6 segment. The stress distribution on the cortical bone surface was more uniform in the porous cage groups, leading to increased average stress values. The gyroid, Schwarz, and BCC cage groups showed higher average stress on the C5 cortical bone. The average stress on the surface of porous cages was higher than that on the surface of high-density cages, with the greatest difference observed under the lateral bending condition. The BCC cage demonstrated favourable mechanical stability. CONCLUSION: The new porous cervical cages satifies requirements of low rigidity and serve as a favourable biological scaffold for bone ingrowth. This study provides valuable insights for the development of next-generation orthopaedic medical devices.

Assessment about bioindicator capacity of acrocarpous moss Campylopus schmidii exposed to abandoned pyritic tailings.

Terrestrial mosses are promising species to study concerning metal deposition, absorption, and soil fertility as moss biocrusts. However, acrocarpous moss, as a kind of terrestrial mosses, has not yet been well understood, both in environmental monitoring and ecological application, especially exposed to an abandoned pyrite mining. Herein, we investigated the concentrations of different heavy metals in soil underlying acrocarpous moss Campylopus schmidii at three distances from an abandoned pyrite mine tailings (0.5, 1, 2 km) by sampling analysis, as well as the accumulation properties of heavy metals in different parts of mosses and soil nutrients under intact mosses and moss-free layers. The results indicated that the soil we researched was heavily polluted by Cr, Cu, and Cd, which was 4.46, 4.18, and 1.77 times higher than the standard of risk screening values for soil environment quality in China. And there was a marked difference in the concentrations and distribution of heavy metals in mosses, with higher concentrations of Cr, Cu, Ni and Pb mainly in the ageing parts. In addition, mosses can effectively promote soil fertility. Compared with the bare soil without the moss layer, the total organic matter and total potassium concentrations of the soil covered by the intact moss layer were significantly increased, by 113.91% and 186.08% respectively. Correlation analysis indicated that similar pollution sources for Zn, Cd, Cu, and Pb, and the concentrations of these heavy metals in soil connected with the distance from the source of pollution. Overall, we expected that these findings could assess the greater potential of single native dominant moss species C.schmidii to act as biomonitors in specific pyrite mine tailings characterized by barren soil with strong acids (pH < 4.0) and polymetallic pollution. Meanwhile, our results revealed may serve as a possibility reference for similar areas and is recommended for developing a vegetative cover utilizing local acrocarpous mosses to achieve greening of degraded tailings in the future, as well as environmental management and protection.

DIA-Based Proteomic Analysis of Plasma Protein Profiles in Patients with Severe Acute Pancreatitis.

Acute pancreatitis (AP) is a pancreatic inflammatory disease that varies greatly in course and severity. To further the understanding of the pathology of AP, we carried out data-independent acquisition-based proteomic analyses using proteins extracted from the plasma of patients with severe acute pancreatitis (SAP) (experimental group) and healthy volunteers (control group). Compared to the control group, there were 35 differentially expressed proteins (DEPs) in the plasma of patients with SAP. Of those, the expression levels for 6 proteins were significantly increased, and 29 proteins were significantly decreased. Moreover, six candidate biomarkers-VWF, ORM2, CD5L, CAT, IGLV3-10, and LTF-were matched as candidate biomarkers of the disease severity of AP. The area under the receiver operating characteristic of 0.903 (95% CI: 0.839, 0.967) indicated that this combination of these six candidate biomarkers had a good prediction accuracy for predicting the severity of AP. Our study provides specific DEPs that may be useful in the diagnosis and prognosis of SAP, which suggests new theoretical bases for the occurrence and development of SAP and offers potential novel treatment strategies for SAP.

Factors associated with public attitudes towards persons with disabilities: a systematic review.

OBJECTIVE: The aim of this review is to identify and summarize factors that are associated with public attitudes towards people with various disabilities systematically. METHODS: An electronic search of three databases was performed (Medline, EMBASE and Cochrane) covering the period from 1950 to present. A comprehensive search strategy was developed and the lists of citations were screened for potential eligible studies. Only quantitative studies using valid measurements were included, and the methodological quality of included studies was appraised based on three criteria (sample, measurement, analysis) by two independent reviewers. RESULTS: The initial electronic search yielded 995 articles after duplicates removed, and 27 studies met the eligibility criteria were included in the study. Three categories of the factors were found to be associated with the public attitudes, which are related to the attitude provider, disabled people, and society respectively. Specifically, the more people know about disabilities, the more likely they were to have positive attitude; and the frequency and quality of the contact with the disabled are also proved to be influential to the attitudes. Meanwhile, the type of disability is also closely correlated to the public's attitude towards the disabilities. CONCLUSION: People's knowledge of the disability and their contact with individuals with disabilities are the main influential factors in public attitudes towards persons with disabilities.

Translation, Cross-Culture Adaptation and Measurement Properties of the Simplified Chinese Version of the Osteoarthritis Knowledge Scale.

OBJECTIVE: The aim of this study was to culturally adapt the Osteoarthritis Knowledge Scale (OAKS) into a simplified Chinese version and to evaluate its measurement properties among Chinese-speaking populations. METHODS: The OAKS scale was translated and cross-culturally adapted following COSMIN guidelines. A total of 368 participants were recruited, including individuals with knee or hip osteoarthritis (OA), healthcare professionals, and individuals without OA. The study evaluated the scale's internal consistency, test-retest reliability, and factor structure using Exploratory Factor Analysis (EFA). RESULTS: The translated OAKS scale demonstrated acceptable levels of reliability across different groups. Cronbach's alpha for the KOAKS was 0.70 overall, with subgroup values ranging from 0.70 to 0.71. For the HOAKS, Cronbach's alpha was 0.77 overall, with subgroup values ranging from 0.63 to 0.72. Test-retest reliability showed high Intraclass Correlation Coefficient (ICC) values for both scales (KOAKS: ICC = 0.960; HOAKS: ICC = 0.982). EFA revealed that the overall OAKS explained a substantial portion of the variance (60.51%), with KOAKS accounting for 59.35% and HOAKS accounting for 54.68% of the variance. CONCLUSION: The simplified Chinese version of the OAKS is a reliable tool for assessing OA knowledge among Chinese-speaking patients, healthcare professionals, and the general public. The findings highlight the necessity for continuous improvement and adaptation of educational interventions to address specific knowledge gaps within the Chinese context.

Participation of Children With Autism During COVID-19: The Role of Maternal Participation.

BACKGROUND: Little is known about participation during adverse times. OBJECTIVES: This study described participation of children with autism aged 6 to 13 during COVID-19 pandemic and examined the extent to which child factors, mother's own participation, and environmental barriers/supports explain child participation in home and community. METHOD: A total of 130 mothers completed the Participation and Environment Measure for Children and Youth, the Health Promoting Activities Scale, functional issues checklist, and sociodemographic questionnaire. RESULTS: Children's participation frequency and involvement were significantly higher at home than in the community. In both settings, mothers desired change in 71% of activities. Multiple regression models indicated that child's age and mother's participation frequency significantly explained child's home involvement (R2 = 21%), where mother's participation (frequency) had a unique contribution (ΔR2 = 10.4%) at home but not in the community. CONCLUSION: Findings imply the importance of maternal participation to child's participation at home and suggest redirecting attention for enhancing family participation as a whole.

Enhancing high-efficient cadmium biosorption of Escherichia coli via cell surface displaying metallothionien CUP1.

Cadmium (Cd) is one of the common heavy metal pollutants in soil, which can induce various diseases and pose a serious threat to human health. Metallothioneins (MTs) are well-known for their excellent metal binding ability due to a high content of cysteine, which has great potential for heavy metal chelation. In this study, we used the Escherichia coli (E. coli) surface display system LPP-OmpA to construct a recombinant plasmid pBSD-LCF encoding LPP-OmpA-CUP1-Flag fusion protein. Then we displayed the metallothionein CUP1 from Saccharomyces cerevisiae on E. coli DH5α surface for Cd removing. The feasibility of surface display of metallothionein CUP1 in recombinant E. coli DH5α (pBSD-LCF) by Lpp-OmpA system was proved by flow cytometry and western blot analysis, and the specificity of the fusion protein in the recombinant strain was also verified. The results showed that the Cd2+ resistance capacity of DH5α (pBSD-LCF) was highly enhanced by about 200%. Fourier-transform infrared spectroscopy showed that sulfhydryl and sulfonyl groups were involved in Cd2+ binding to cell surface of DH5α (pBSD-LCF). Meanwhile, Cd removal rate by DH5α (pBSD-LCF) was promoted to 95.2%. Thus, the recombinant strain E. coli DH5α (pBSD-LCF) can effectively chelate environmental metals, and the cell surface expression of metallothionein on E. coli can provide new ideas and directions for heavy metals remediation.

Integrated analysis of circRNA- related ceRNA network targeting neuroinflammation in medial temporal lobe epilepsy.

BACKGROUND: medial temporal lobe epilepsy (mTLE) is among the most common types of temporal lobe epilepsy (TLE) ，it is generally resistant to drug treatment, which significantly impacts the quality of life and treatment. Research on novel therapeutic approaches for mTLE has become a current focus. Our study aims to construct and analyze a competing endogenous RNA (ceRNA) network that targets neuroinflammation using publicly available data, which may offer a novel therapeutic approach for mTLE. METHODS: we utilized the R package to analyze GSE186334 downloaded from Gene Expression Omnibus database, subsequently constructing and identifying hub network within the ceRNA network using public databases. Lastly, we validated the expressions and interactions of some nodes within the hub ceRNA network in Sombati cell model. RESULTS: our transcriptome analysis identified 649 differentially expressed (DE) mRNAs (273 up-regulated, 376 down-regulated) and 36 DE circRNAs (11 up-regulated, 25 down-regulated) among mTLE patients. A total of 23 candidate DE mRNAs associated with neuroinflammation were screened, and two ceRNA networks were constructed. A hub network was further screened which included 3 mRNAs, 22 miRNAs, and 11 circRNAs. Finally, we confirmed the hsa-miR-149-5p is crucial in the regulatory effect of hsa_circ_0005145 on IL - 1α in the hub network. CONCLUSIONS: In summary, our study identified a hub ceRNA network and validated a potential circRNA-miRNA-mRNA axis targeting neuroinflammation. The results of our research may serve as a potential therapeutic target for mTLE.

Origami-inspired highly stretchable and breathable 3D wearable sensors for in-situ and online monitoring of plant growth and microclimate.

The emerging wearable plant sensors demonstrate the capability of in-situ measurement of physiological and micro-environmental information of plants. However, the stretchability and breathability of current wearable plant sensors are restricted mainly due to their 2D planar structures, which interfere with plant growth and development. Here, origami-inspired 3D wearable sensors have been developed for plant growth and microclimate monitoring. Unlike 2D counterparts, the 3D sensors demonstrate theoretically infinitely high stretchability and breathability derived from the structure rather than the material. They are adjusted to 100% and 111.55 mg cm-2·h-1 in the optimized design. In addition to stretchability and breathability, the structural parameters are also used to control the strain distribution of the 3D sensors to enhance sensitivity and minimize interference. After integrating with corresponding sensing materials, electrodes, data acquisition and transmission circuits, and a mobile App, a miniaturized sensing system is produced with the capability of in-situ and online monitoring of plant elongation and microclimate. As a demonstration, the 3D sensors are worn on pumpkin leaves, which can accurately monitor the leaf elongation and microclimate with negligible hindrance to plant growth. Finally, the effects of the microclimate on the plant growth is resolved by analyzing the monitored data. This study would significantly promote the development of wearable plant sensors and their applications in the fields of plant phenomics, plant-environment interface, and smart agriculture.

Selective Upcycling of Polyethylene over Ru/H-ZSM-5 Bifunctional Catalyst into High-Quality Liquid Fuel at Mild Conditions.

It has been known that plastics with undegradability and long half-times have caused serious environmental and ecological issues. Considering the devastating effects, the development of efficient plastic upcycling technologies with low energy consumption is absolutely imperative. Catalytic hydrogenolysis of single-use polyethylene over Ru-based catalysts to produce high-quality liquid fuel has been one of the current top priority strategies, but it is restricted by some tough challenges, such as the tendency towards methanation resulting from terminal C-C cleavage. Herein, we introduced Ru nanoparticles supported on hollow ZSM-5 zeolite (Ru/H-ZSM-5) for hydrocracking of high-density polyethylene (HDPE) under mild reaction conditions. The implication of experimental results is that the 1Ru/H-ZSM-5 (~1 wt % Ru) acted as an effective and reusable bifunctional catalyst providing higher conversion rate (82.53 %) and liquid fuel (C5-C21) yield (62.87 %). Detailed characterization demonstrated that the optimal performance in hydrocracking of PE could be attributed to the moderate acidity and appropriate positively charged Ru species resulting from the metal-zeolite interaction. This work proposes a promising catalyst for plastic upcycling and reveals its structure-performance relationship, which has guiding significance for catalyst design to improve the yield of high-value liquid fuels.

Mfn2R364W, Mfn2G176S, and Mfn2H165R mutations drive Charcot-Marie-Tooth type 2A disease by inducing apoptosis and mitochondrial oxidative phosphorylation damage.

Charcot-Marie-Tooth type 2A (CMT2A) is a single-gene motor sensory neuropathy caused by Mfn2 mutation. It is generally believed that CMT2A involves mitochondrial fusion disruption. However, how Mfn2 mutation mediates the mitochondrial membrane fusion loss and its further pathogenic mechanisms remain unclear. Here, in vivo and in vitro mouse models harboring the Mfn2R364W, Mfn2G176S and Mfn2H165R mutations were constructed. Mitochondrial membrane fusion and fission proteins analysis showed that Mfn2R364W, Mfn2G176S, and Mfn2H165R/+ mutations maintain the expression of Mfn2, but promote Drp1 upregulation and Opa1 hydrolytic cleavage. In Mfn2H165R/H165R mutation, Mfn2, Drp1, and Opa1 all play a role in inducing mitochondrial fragmentation, and the mitochondrial aggregation is affected by Mfn2 loss. Further research into the pathogenesis of CMT2A showed these three mutations all induce mitochondria-mediated apoptosis, and mitochondrial oxidative phosphorylation damage. Overall, loss of overall fusion activity affects mitochondrial DNA (mtDNA) stability and causes mitochondrial loss and dysfunction, ultimately leading to CMT2A disease. Interestingly, the differences in the pathogenesis of CMT2A between Mfn2R364W, Mfn2G176S, Mfn2H165R/+ and Mfn2H165R/H165R mutations, including the distribution of Mfn2 and mitochondria, the expression of mitochondrial outer membrane-associated proteins (Bax, VDAC1 and AIF), and the enzyme activity of mitochondrial complex I, are related to the expression of Mfn2.

Recent trends in phenol synthesis by photocatalytic oxidation of benzene.

Phenol is an important intermediate for manufacturing chemical products in industry. In recent decades, phenol synthesis by one-pot oxidation of benzene has aroused tremendous interest in phenol synthesis due to the enormous energy consumption of the three-step cumene method in the industry. Photocatalysis is promising for the selective conversion of benzene to phenol because it can proceed under mild reaction conditions. However, overoxidation of phenol by photocatalysts with high oxidation ability decreases the yield and selectivity, which is the major limiting factor. Thus, increasing the phenol formation efficiency plays a crucial role in photocatalytic systems for benzene oxidation. In this context, selective photocatalytic benzene oxidation over several types of photocatalytic systems has been developed rapidly in the past few years. In this perspective, current homogeneous and heterogeneous photocatalytic systems for this reaction have been reviewed systematically first. Then, an overview of some strategies from the last decade for increasing phenol selectivity has been provided. In the end, a summary and outlook on the challenges and future directions in the research field are included in this perspective, which would be of great interest for further improving the selectivity of the photocatalytic benzene oxidation reaction.

An integrative analysis of genotype-phenotype correlation in Charcot Marie Tooth type 2A disease with MFN2 variants: A case and systematic review.

Dominant genetic variants in the mitofusin 2 (MFN2) gene lead to Charcot-Marie-Tooth type 2A (CMT2A), a neurodegenerative disease caused by genetic defects that directly damage axons. In this study, we reported a proband with a pathogenic variant in the GTPase domain of MFN2, c.494A > G (p.His165Arg). To date, at least 184 distinct MFN2 variants identified in 944 independent probands have been reported in 131 references. However, the field of medical genetics has long been challenged by how genetic variation in the MFN2 gene is associated with disease phenotypes. Here, by collating the MFN2 variant data and patient clinical information from Leiden Open Variant Database 3.0, NCBI clinvar database, and available related references in PubMed, we determined the mutation frequency, age of onset, sex ratio, and geographical distribution. Furthermore, the results of an analysis examining the relationship between variants and phenotypes from multiple genetic perspectives indicated that insertion and deletions (indels), copy number variants (CNVs), duplication variants, and nonsense mutations in single nucleotide variants (SNVs) tend to be pathogenic, and the results emphasized the importance of the GTPase domain to the structure and function of MFN2. Overall, three reliable classification methods of MFN2 genotype-phenotype associations provide insights into the prediction of CMT2A disease severity. Of course, there are still many MFN2 variants that have not been given clear clinical significance, which requires clinicians to make more accurate clinical diagnoses.

Decreasing predictable DNA off-target effects and narrowing editing windows of adenine base editors by fusing human Rad18 protein variant.

Adenine base editors, enabling targeted A-to-G conversion in genomic DNA, have enormous potential in therapeutic applications. However, the currently used adenine base editors are limited by wide editing windows and off-target effects in genetic therapy. Here, we report human e18 protein, a RING type E3 ubiquitin ligase variant, fusing with adenine base editors can significantly improve the preciseness and narrow the editing windows compared with ABEmax and ABE8e by diminishing the abundance of base editor protein. As a proof of concept, ABEmax-e18 and ABE8e-e18 dramatically decrease Cas9-dependent and Cas9-independent off-target effects than traditional adenine base editors. Moreover, we utilized ABEmax-e18 to establish syndactyly mouse models and achieve accurate base conversion at human PCSK9 locus in HepG2 cells which exhibited its potential in genetic therapy. Furthermore, a truncated version of base editors-RING (ABEmax-RING or AncBE4max-RING), which fusing the 63 amino acids of e18 protein RING domain to the C terminal of ABEmax or AncBE4max, exhibited similar effect compared to ABEmax-e18 or AncBE4max-e18.In summary, the e18 or RING protein fused with base editors strengthens the precise toolbox in gene modification and maybe works well with various base editing tools with a more applicable to precise genetic therapies in the future.

[Mechanism of Nampt gene expression regulation].

Nicotinamide phosphoribosyltransferase (NAMPT), also known as pre-B cell colony-enhancing factor (PBEF) or visfatin, is a crucial rate-limiting enzyme of NAD biosynthetic pathway. It may affect the metabolism, inflammatory response, cell proliferation, differentiation, and apoptosis, especially the aging and other physiological progresses through regulating NAD biosynthesis and nonenzyme routes in the organisms and cells. This review mainly focuses on recent progresses in the expression modulation and feedback regulation of Nampt gene.