Establishment of an AAV Reverse Infection-Based Array

Background: The development of a convenient high-throughput gene transduction approach is critical for biological screening. Adeno-associated virus (AAV) vectors are broadly used in gene therapy studies, yet their applications in in vitro high-throughput gene transduction are limited. Principal Findings: We established an AAV reverse infection (RI)-based method in which cells were transduced by quantified recombinant AAVs (rAAVs) pre-coated onto 96-well plates. The number of pre-coated rAAV particles and number of cells loaded per well, as well as the temperature stability of the rAAVs on the plates, were evaluated. As the first application of this method, six serotypes or hybrid serotypes of rAAVs (AAV1, AAV2, AAV5/5, AAV8, AAV25 m, AAV28 m) were compared for their transduction efficiencies using various cell lines, including BHK21, HEK293, BEAS-2BS, HeLaS3, Huh7, Hepa1-6, and A549. AAV2 and AAV1 displayed high transduction efficiency; thus, they were deemed to be suitable candidate vectors for the RI-based array. We next evaluated the impact of sodium butyrate (NaB) treatment on rAAV vectormediated reporter gene expression and found it was significantly enhanced, suggesting that our system reflected the biological response of target cells to specific treatments. Conclusions/Significance: Our study provides a novel method for establishing a highly efficient gene transduction arra

Diversification and Molecular Evolution of ATOH8, a Gene Encoding a bHLH Transcription Factor

ATOH8 is a bHLH domain transcription factor implicated in the development of the nervous system, kidney, pancreas, retina and muscle. In the present study, we collected sequence of ATOH8 orthologues from 18 vertebrate species and 24 invertebrate species. The reconstruction of ATOH8 phylogeny and sequence analysis showed that this gene underwent notable divergences during evolution. For those vertebrate species investigated, we analyzed the gene structure and regulatory elements of ATOH8. We found that the bHLH domain of vertebrate ATOH8 was highly conserved. Mammals retained some specific amino acids in contrast to the non-mammalian orthologues. Mammals also developed another potential isoform, verified by a human expressed sequence tag (EST). Comparative genomic analyses of the regulatory elements revealed a replacement of the ancestral TATA box by CpG-islands in the eutherian mammals and an evolutionary tendency for TATA box reduction in vertebrates in general. We furthermore identified the region of the effective promoter of human ATOH8 which could drive the expression of EGFP reporter in the chicken embryo. In the opossum, both the coding region and regulatory elements of ATOH8 have some special features, such as the unique extended C-terminus encoded by the third exon and absence of both CpG islands and TATA elements in the regulatory region. Our gene mapping data showed that in human, ATOH8 was hosted in one chromosome which is a fusion product of two orthologous chromosomes in non-human primates. This unique chromosomal environment of human ATOH8 probably subjects its expression to the regulation at chromosomal level. We deduce that the great interspecific differences found in both ATOH8 gene sequence and its regulatory elements might be significant for the fine regulation of its spatiotemporal expression and roles of ATOH8, thus orchestrating its function in different tissues and organisms

Methylprednisolone as Adjunct to Endovascular Thrombectomy for Large-Vessel Occlusion Stroke

Importance It is uncertain whether intravenous methylprednisolone improves outcomes for patients with acute ischemic stroke due to large-vessel occlusion (LVO) undergoing endovascular thrombectomy. Objective To assess the efficacy and adverse events of adjunctive intravenous low-dose methylprednisolone to endovascular thrombectomy for acute ischemic stroke secondary to LVO. Design, Setting, and Participants This investigator-initiated, randomized, double-blind, placebo-controlled trial was implemented at 82 hospitals in China, enrolling 1680 patients with stroke and proximal intracranial LVO presenting within 24 hours of time last known to be well. Recruitment took place between February 9, 2022, and June 30, 2023, with a final follow-up on September 30, 2023.InterventionsEligible patients were randomly assigned to intravenous methylprednisolone (n = 839) at 2 mg/kg/d or placebo (n = 841) for 3 days adjunctive to endovascular thrombectomy. Main Outcomes and Measures The primary efficacy outcome was disability level at 90 days as measured by the overall distribution of the modified Rankin Scale scores (range, 0 [no symptoms] to 6 [death]). The primary safety outcomes included mortality at 90 days and the incidence of symptomatic intracranial hemorrhage within 48 hours. Results Among 1680 patients randomized (median age, 69 years; 727 female [43.3%]), 1673 (99.6%) completed the trial. The median 90-day modified Rankin Scale score was 3 (IQR, 1-5) in the methylprednisolone group vs 3 (IQR, 1-6) in the placebo group (adjusted generalized odds ratio for a lower level of disability, 1.10 [95% CI, 0.96-1.25]; P = .17). In the methylprednisolone group, there was a lower mortality rate (23.2% vs 28.5%; adjusted risk ratio, 0.84 [95% CI, 0.71-0.98]; P = .03) and a lower rate of symptomatic intracranial hemorrhage (8.6% vs 11.7%; adjusted risk ratio, 0.74 [95% CI, 0.55-0.99]; P = .04) compared with placebo. Conclusions and Relevance Among patients with acute ischemic stroke due to LVO undergoing endovascular thrombectomy, adjunctive methylprednisolone added to endovascular thrombectomy did not significantly improve the degree of overall disability.Trial RegistrationChiCTR.org.cn Identifier: ChiCTR210005172

Torque Ripple Suppression of Brushless DC Motor Drive System Based on Improved Harmonic Injection Active Disturbance Rejection Control

The positioning accuracy and speed stability of the brushless DC motor (BLDC motor), as the drive element of the optomechanically scanned system (OMSS), are closely interrelated to the final imaging quality of the system. Active disturbance rejection control (ADRC) with strong anti-interference ability, fast response and good robustness is one of the extensively used control strategies. However, the performance of ADRC working in a complicated environment will be limited due to the controller structure, parameter tuning and the influence of multi-source nonlinear disturbance. Therefore, an improved ADRC method is proposed, which can switch between ‘point-to-point control mode’ and ‘stable speed control mode’ according to the system requirements. To further suppress the torque ripple and improve the control performance of the system, an improved harmonic injection scheme is added, and the parameters of improved ADRC are tuned by a slime mould algorithm based on a Levy flight operator (LF-SMA). The stability of the proposed ADRC is proved by Lyapunov stability theory. The experimental results show that the proposed control scheme could be available to reduce the torque ripple of the system

Organosilicon Based Electrolytes for Lithium-Ion Batteries

Organosilicon compounds have attracted considerable interest as electrolytes for lithium-ion batteries because they are nontoxic, nonflammable, as well as have lower glass transition temperatures, lower vapor pressure and higher flash point than commerical alkyl carbonates. These compounds can improve the electrochemical performances and safety of lithium-ion batteries when used as electrolyte solvents or additives in the electrolytes. In this paper, the recent advances of organosilicon compounds both as electrolyte solvents and additives are reviewed. Organosilicon electrolytes containing ethylene oxide (EO) substituents with or without carbon spacer between silicon atom and EO unit are specially remarked as safe electrolytes. Organosilicon compounds as functional additives are also introduced in terms of the capabilities of passive film formation, flame-retardant, and acid/water scavenger

Development of a Smart Clinical Bluetooth Thermometer Based on an Improved Low-Power Resistive Transducer Circuit

Smart sensors have been used in many engineering monitoring and control applications. This work focuses on the development of a new type of clinical Bluetooth thermometer, based on an improved low-power resistive transducer circuit. Most existing resistive transducers use relatively complicated circuits with higher cost and power consumption. To tackle these problems, especially in real applications, an improved low-power resistive transducer circuit is proposed in this work and is used to develop smart Bluetooth thermometers. The parameters of the resistive transducer circuit are selected by quantitative analysis and optimization to improve the performance of the low-power resistive transducer circuit. The effectiveness of the proposed design technology was verified by tests. The temperature measurement error of the new smart Bluetooth thermometer is less than 0.1 °C, which can not only meet the clinical use requirements but also has lower cost and power consumption

A Novel Aminoalkylsilane Compound with Oligo(ethylene oxide) Units as Effective Additive for Improving Cyclability of Lithium-ion Batteries

A new aminoalkylsilane compound, ((2-(2-(N,N-dimethylamino)ethoxy)ethoxy) methyl)trimethylsilane (TMSC1N2) based on the oligo(ethylene oxide) chain end-capped with organosilicon functional group and alkylamine group on each end, was introduced as an electrolyte additive for lithium-ion batteries. Electrochemical performances of different volume ratios of TMSC1N2 in the baseline electrolyte were conducted through cyclic voltammetry, electrochemical impedance spectroscopy and galvanostatic charge/discharge tests of lithium-ion batteries. With adding 5 vol.% TMSC1N2 to the baseline electrolyte (1 mol/L LiPF6 in ethylene carbonate and diethyl carbonate (EC:DEC = 1: 1, in volume)), the capacity retention of LiFePO4/Li cells could be significantly improved from 74.7% to 90.8% after 130 cycles. Furthermore, TMSC1N2 showed good compatibility with graphite electrode and would not deteriorate the electrochemical performance of graphite/Li anode cells. These data suggested that TMSC1N2 could be utilized as an effective additive for lithium-ion batteries