Cardio-respiratory and phenotypic rescue of dystrophin/utrophin-deficient mice by combination therapy.

Duchenne muscular dystrophy (DMD) is a systemic progressive muscular disease caused by frame-disrupting mutations in the DMD gene. Although exon-skipping antisense oligonucleotides (AOs) are clinically approved and can correct DMD, insufficient muscle delivery limits efficacy. If AO activity can be enhanced by safe dietary supplements, clinical trials for efficacy can be undertaken rapidly to benefit patients. We showed previously that intravenous glycine enhanced phosphorodiamidate morpholino oligomer (PMO) delivery to peripheral muscles in mdx mice. Here, we demonstrate that the combination of oral glycine and metformin with intravenous PMO enhances PMO activity, dystrophin restoration, extends lifespan, and improves body-wide function and phenotypic rescue of dystrophin /utrophin double knock-out (DKO) mice without any overt adverse effects. The DKO mice treated with the combination without altering the approved administration protocol of PMO show improved cardio-respiratory and behavioral functions. Metformin and glycine individually are ineffective in DMD patients, but the combination of PMO with clinically-approved oral glycine and metformin might improve the efficacy of the treatment also in DMD patients. Our data suggest that this combination therapy might be an attractive therapy for DMD and potentially other muscle diseases requiring systemic treatment with AOs.

Investigating potential biomarkers of acute pancreatitis in patients with a BMI>30 using Mendelian randomization and transcriptomic analysis.

BACKGROUND: Acute pancreatitis (AP) has become a significant global health concern, and a high body mass index (BMI) has been identified as a key risk factor exacerbating this condition. Within this context, lipid metabolism assumes a critical role. The complex relationship between elevated BMI and AP, mediated by lipid metabolism, markedly increases the risk of complications and mortality. This study aimed to accurately define the correlation between BMI and AP, incorporating a comprehensive analysis of the interactions between individuals with high BMI and AP. METHODS: Mendelian randomization (MR) analysis was first applied to determine the causal relationship between BMI and the risk of AP. Subsequently, three microarray datasets were obtained from the GEO database. This was followed by an analysis of differentially expressed genes and the application of weighted gene coexpression network analysis (WGCNA) to identify key modular genes associated with AP and elevated BMI. Functional enrichment analysis was then performed to shed light on disease pathogenesis. To identify the most informative genes, machine learning algorithms, including Random Forest (RF), Support Vector Machine-Recursive Feature Elimination (SVM-RFE), and Least Absolute Shrinkage and Selection Operator (LASSO), were employed. Subsequent analysis focused on the colocalization of the Quantitative Trait Loci (eQTL) data associated with the selected genes and Genome-Wide Association Studies (GWAS) data related to the disease. Preliminary verification of gene expression trends was conducted using external GEO datasets. Ultimately, the diagnostic potential of these genes was further confirmed through the development of an AP model in mice with a high BMI. RESULTS: A total of 21 intersecting genes related to BMI>30, AP, and lipid metabolism were identified from the datasets. These genes were primarily enriched in pathways related to cytosolic DNA sensing, cytokine‒cytokine receptor interactions, and various immune and inflammatory responses. Next, three machine learning techniques were utilized to identify HADH as the most prevalent diagnostic gene. Colocalization analysis revealed that HADH significantly influenced the risk factors associated with BMI and AP. Furthermore, the trend in HADH expression within the external validation dataset aligned with the trend in the experimental data, thus providing a preliminary validation of the experimental findings.The changes in its expression were further validated using external datasets and quantitative real-time polymerase chain reaction (qPCR). CONCLUSION: This study systematically identified HADH as a potential lipid metabolism-grounded biomarker for AP in patients with a BMI>30.

Voltage-gated potassium channels KCNQs: Structures, mechanisms, and modulations.

KCNQ (Kv7) channels are voltage-gated, phosphatidylinositol 4,5-bisphosphate- (PIP2-) modulated potassium channels that play essential roles in regulating the activity of neurons and cardiac myocytes. Hundreds of mutations in KCNQ channels are closely related to various cardiac and neurological disorders, such as long QT syndrome, epilepsy, and deafness, which makes KCNQ channels important drug targets. During the past several years, the application of single-particle cryo-electron microscopy (cryo-EM) technique in the structure determination of KCNQ channels has greatly advanced our understanding of their molecular mechanisms. In this review, we summarize the currently available structures of KCNQ channels, analyze their special voltage gating mechanism, and discuss their activation mechanisms by both the endogenous membrane lipid and the exogenous synthetic ligands. These structural studies of KCNQ channels will guide the development of drugs targeting KCNQ channels.

[Human muscle fatigue monitoring method and its application for exoskeleton interactive control].

Aiming at the human-computer interaction problem during the movement of the rehabilitation exoskeleton robot, this paper proposes an adaptive human-computer interaction control method based on real-time monitoring of human muscle state. Considering the efficiency of patient health monitoring and rehabilitation training, a new fatigue assessment algorithm was proposed. The method fully combined the human neuromuscular model, and used the relationship between the model parameter changes and the muscle state to achieve the classification of muscle fatigue state on the premise of ensuring the accuracy of the fatigue trend. In order to ensure the safety of human-computer interaction, a variable impedance control algorithm with this algorithm as the supervision link was proposed. On the basis of not adding redundant sensors, the evaluation algorithm was used as the perceptual decision-making link of the control system to monitor the muscle state in real time and carry out the robot control of fault-tolerant mechanism decision-making, so as to achieve the purpose of improving wearing comfort and improving the efficiency of rehabilitation training. Experiments show that the proposed human-computer interaction control method is effective and universal, and has broad application prospects.

Ultrafast Time-resolved Polarization-dependent Investigations on the Dynamics in the A ˜ 2 B2 State of NO2 Molecules.

We perform time-resolved polarization-dependent study on ultrafast dynamics in the A ˜ 2 B2 state of NO2 . A linearly-polarized 400 nm femtosecond laser is used to resonantly pump NO2 to its first excited state A ˜ 2 B2 , and the time-dependent ionic yields produced via strong field ionization at 800 nm are measured under different laser polarizations. The yield ratios measured with the lasers perpendicular and parallel to each other first decrease and then increases as the wave packet evolves on the excited state, with a minimum ratio at 180 fs delay time, which can be attributed to the evolution time in the A ˜ 2 B2 state. The behavior of the time-resolved ionization in elliptically polarized laser field is also investigated and discussed.

Isolation and Synthesis of Azuriaplysins A and B, Bromoditerpenes with an α-Methylene Carbonyl from the Sea Hare Aplysia kurodai.

New bromoditerpenes having an α-methylene carbonyl structure, azuriaplysins A (1) and B (2), were isolated from the sea hare Aplysia kurodai. Their relative stereostructures were determined based on one- and two-dimensional NMR spectroscopic analysis. In addition, the absolute stereostructures were determined by the total synthesis of both enantiomers of azuriaplysins A (1) and B (2), the key points of which were bromocyclization of farnesol and optical resolution of a key intermediate. Azuriaplysin B (2) and its enantiomer exhibited moderate cytotoxicity against HeLa S3 cells.

Design of non-equal-strut stent hoops for structural optimization of thoracic aortic stent-grafts.

BACKGROUND: Endovascular aortic aneurysm repair (EVAR) with stent-grafts is used widely for the treatment of thoracic aortic aneurysms (TAA). Inappropriate design of stent-grafts may lead to complications such as endoleak, stent-graft migration and new entries, causes of which may be inappropriate radial support force or insufficient longitudinal flexibility of the stent-grafts. MATERIAL AND METHODS: To improve the mechanical performance of the stent-grafts, a type of non-equal-strut stent hoops was proposed, and the influence of structural parameters on the mechanical performance was studied. RESULTS: Results of numerical simulation and physical experiments show that by using the proposed non-equal-strut stent hoops, radial support force and longitudinal flexibility of stent-grafts can be reconciled and balanced. CONCLUSION: Results of this study could be used to facilitate radial force control and longitudinal flexibility enhancement in the design of aortic stent-grafts.

Strong Field Ionization-Photofragmentation on Ultrafast Evolution of Electronic States of Toluene Cations.

Ultrafast time-resolved strong field ionization-photofragmentation (SFI-PF) has emerged as a useful method for investigation of dynamics of molecular cations. Here we perform a SFI-PF study on the electronic states of toluene cations. By measuring the ion yields as a function of delay time, we obtain the transients of both parent and daughter ions, which show ultrafast decays and out-of-phase oscillations. The results provide the first experimental evidence of D1-D0 ultrafast relaxation of toluene cations occurring in about 530 fs and indicate coincident resonance between the vibrational states in D1 and D0 leading to oscillations with a period of about 2.05 ps. Our study should shed some light on the ultrafast photochemistry involving complex molecular cations.

Characteristics of Myelogram in Patients with Extraocular Metastatic Retinoblastoma and Morphological Analysis of Tumor Cells in Bone Marrow and Cerebrospinal Fluid.

BACKGROUND: Retinoblastoma (RB) is a common intraocular malignant tumor in infants and young children. However, reports on the morphological descriptions of RB tumor cells from native and foreign scholars are rare. OBJECTIVES: To investigate the myelogram characteristics of RB with extraocular tumor extension and the morphological characteristics of tumor cells in the bone marrow and cerebrospinal fluid. METHODS: For the period from May 2011 to February 2015, we analyzed clinical data on 18 patients in our hospital diagnosed as having metastatic RB in the extraocular and other distant regions associated with clear bone marrow metastasis. The morphology of tumor cells in the bone marrow and cerebrospinal fluid was retrospectively analyzed after staining with Wright-Giemsa stain. A summary of the cytological characteristics was also presented. RESULTS: RB tumor cells in the bone marrow and cerebrospinal fluid not only appeared as aggregated clumps, but were distributed in a scattered manner. The tumor cells may present different characteristic morphologies in different cases, with different tumor cell smears from the same tumor mass even showing different features. According to the degree of tumor metastasis, changes in myelogram were significantly different. CONCLUSION: The tumor cells of RB patients show unique morphological characteristics in the bone marrow and cerebrospinal fluid. Therefore, correct identification of the cells is of great value in the diagnosis, staging, and prognosis of RB.

[Convolutional neural network based on temporal-spatial feature learning for motor imagery electroencephalogram signal decoding].

With the advantage of providing more natural and flexible control manner, brain-computer interface systems based on motor imagery electroencephalogram (EEG) have been widely used in the field of human-machine interaction. However, due to the lower signal-noise ratio and poor spatial resolution of EEG signals, the decoding accuracy is relative low. To solve this problem, a novel convolutional neural network based on temporal-spatial feature learning (TSCNN) was proposed for motor imagery EEG decoding. Firstly, for the EEG signals preprocessed by band-pass filtering, a temporal-wise convolution layer and a spatial-wise convolution layer were respectively designed, and temporal-spatial features of motor imagery EEG were constructed. Then, 2-layer two-dimensional convolutional structures were adopted to learn abstract features from the raw temporal-spatial features. Finally, the softmax layer combined with the fully connected layer were used to perform decoding task from the extracted abstract features. The experimental results of the proposed method on the open dataset showed that the average decoding accuracy was 80.09%, which is approximately 13.75% and 10.99% higher than that of the state-of-the-art common spatial pattern (CSP) + support vector machine (SVM) and filter bank CSP (FBCSP) + SVM recognition methods, respectively. This demonstrates that the proposed method can significantly improve the reliability of motor imagery EEG decoding.

Red- and Far-Red-Emitting Zinc Probes with Minimal Phototoxicity for Multiplexed Recording of Orchestrated Insulin Secretion.

Zinc biology, featuring intertwining signaling networks and critical importance to human health, witnesses exciting opportunities in the big data era of physiology. Here, we report a class of red- and far-red-emitting Zn2+ probes with Kd values ranging from 190 nM to 74 µM, which are particularly suitable for real-time monitoring the high concentration of Zn2+ co-released with insulin during vesicular secretory events. Compared to the prototypical rhodamine-based Zn2+ probes, the new class exploits morpholino auxochromes which eliminates phototoxicity during long-term live recording of isolated islets. A Si-rhodamine-based Zn2+ probe with high turn-on ratio (>100), whose synthesis was enabled by a new route featuring late-stage N-alkylation, allowed simultaneous recording of Ca2+ influx, mitochondrial signal, and insulin secretion in isolated mouse islets. The time-lapse multicolor fluorescence movies and their analysis, enabled by red-shifted Zn2+ and other orthogonal physiological probes, highlight the potential impact of biocompatible fluorophores on the fields of islet endocrinology and system biology.

Putative functional variants of PI3K/AKT/mTOR pathway are associated with knee osteoarthritis susceptibility.

BACKGROUND: Osteoarthritis (OA) is a degenerative musculoskeletal disease which causes joint deformity and pain and finally leads to limb dysfunction. Knee osteoarthritis (KOA) has the highest incidence among all kinds of OA. Strong evidence leads to the understanding that P13K/AKT/mTOR signaling is very important in cartilage degeneration. METHODS: This research sought to understand the association between genetic variation of PI3K/AKT/mTOR genes and KOA susceptibility among Chinese population. All the genetic variants of PI3K/AKT/mTOR pathway were graded and selected using RegulomeDB database, and then, an association study including 278 osteoarthritis patients and 289 controls was conducted. RESULTS: Finally, eight SNPs' genotypes' distributions and susceptibility to KOA were presented. AKT1 rs2498789 was associated with KOA susceptibility in dominate genetic model (AA + GA vs GG) after adjusted for BMI, age, and gender: OR = 1.46, 95% CI: 1.03-2.05, P = .03. PIK3CA rs7646409 was also associated with KOA susceptibility (TC vs TT) after adjusted for BMI, age, and gender: OR = 0.58, 95% CI: 0.36-0.93, P = .02. PIK3CA rs7646409 (TC vs TT) with KOA risk was more significant in age < 60 group (P for heterogeneity was .03). Risk score showed significant association with KOA susceptibility after cumulative analysis (OR = 2.45, 95% CI: 1.35-4.45, P = .003). CONCLUSIONS: This study shows that genetic variation of PI3K/AKT/mTOR is associated with KOA susceptibility in Chinese Han population, indicating that PI3K/AKT/mTOR is very important in KOA pathogenesis.

LncRNA MSC-AS1 promotes osteogenic differentiation and alleviates osteoporosis through sponging microRNA-140-5p to upregulate BMP2.

This study aims to explore the role of lncRNA MSC-AS1/microRNA-140-5p/BMP2 regulatory loop in promoting osteogenic differentiation of BMSCs. BMSCs were isolated from bone marrow of mice. Expression levels of MSC-AS1, microRNA-140-5p and BMP2 during osteogenic differentiation were detected by qRT-PCR. Meanwhile, regulatory effect of MSC-AS1 on osteogenic differentiation was detected through ALP staining and alizarin red staining. The binding sites between microRNA-140-5p and MSC-AS1 as well as between microRNA-140-5p and BMP2 were predicted by TargetScan, which were further confirmed by dual-luciferase reporter gene assay. In addition, protein levels of MSC-AS1/microRNA-140-5p/BMP2 were detected by Western blot. Finally, rescue experiments were conducted to clarify the regulatory effects of MSC-AS1/microRNA-140-5p/BMP2 axis on osteogenic differentiation. MSC-AS1 and BMP2 were found to be remarkably up-regulated during osteogenic differentiation, while microRNA-140-5p was conversely down-regulated. Meanwhile, knockdown of MSC-AS down-regulated expression levels of osteogenesis-associated genes and weakened the mineralization capacity of BMSCs. MicroRNA-140-5p was verified to bind to the 3'UTR of MSC-AS1 and BMP2 genes. Knockdown of MSC-AS1 in BMSCs could reduce the expression of microRNA-140-5p, while knockdown of microRNA-140-5p also down-regulated BMP2 level. In addition, co-silence of MSC-AS1 and microRNA-140-5p reversed the inhibitory effect of MSC-AS1 knockdown on osteogenic differentiation and protein levels of p-Smad1/5/8, RUNX2 and Osterix. MSC-AS1 might promote the osteogenic differentiation of BMSCs through sponging microRNA-140-5p to up-regulate BMP2, thus alleviating the progression of osteoporosis.

A small non-coding RNA facilitates Brucella melitensis intracellular survival by regulating the expression of virulence factor.

Brucella species are the causative agents of brucellosis, a worldwide zoonotic disease that affects a broad range of mammals and causes great economic losses. Small regulatory RNAs (sRNAs) are post-transcriptional regulatory molecules that participate in the stress adaptation and pathogenesis of Brucella. In this study, we characterized the role of a novel sRNA, BSR1141, in the intracellular survival and virulence of Brucella melitensis. The results show that BSR1141 was highly induced during host infections and under in vitro stress situations that simulated the conditions encountered within host phagocytes. In addition, a BSR1141 mutant showed reduced survival both under in vitro stress conditions and in mice, confirming the role of BSR1141 in Brucella intracellular survival. Bioinformatic and experimental approaches revealed that BSR1141 affects the expression of many target genes, including the Brucella virulence component virB2. These data indicate that BSR1141 could influence the expression of virB2, which is important for B. melitensis pathogenesis and intracellular survival. This work provides new insight into the mechanism of adaptation to environmental stress and into the pathogenesis of intracellular pathogens.

Traumatic fractures as a result of motor vehicle collisions in children and adolescents.

PURPOSE: We investigated the incidence and pattern of traumatic fractures resulting from motor vehicle collisions in a population of children and adolescents (≤18 years old) and to determine the risk factors for nerve injury. METHODS: We retrospectively reviewed 734 patients admitted to our university-affiliated hospitals from 2001 to 2010. RESULTS: This study enrolled 498 male (67.8%) and 236 female (32.2%) patients aged 10.9 ± 5.3 years old. The most common injuries were to pedestrians, and the most common fracture sites (438, 59.7%) were to lower extremities (n = 441, 60.0%). A total of 201 (27.4%) patients experienced a nerve injury. Univariate logistic regression analysis showed that age (P = 0.014), lower-extremity (P = 0.000), craniofacial (P = 0.000) and spinal (P = 0.000) fractures were risk factors for nerve injury. Multivariate logistic regression analysis indicated that craniofacial [odds ratio (OR) = 9.003, 95% confidence interval (CI) 5.159-15.711, P = 0.000)] and spinal (experiencedOR = 10.141, 95% CI: 4.649-22.121, P = 0.011) fractures were independent risk factors for nerve injury. CONCLUSIONS: Patients in the 15- to 18-years old group and drivers had the largest sex ratio and highest frequencies of both nerve injury and early complications. Craniofacial and spinal fractures were independent risk factors for nerve injury. It is therefore important to focus on these risk factors to determine the presence of a nerve injury so that early, timely diagnosis and targeted treatment can be provided.

Molecular Cloning, mRNA Expression, and Localization of the G-protein Subunit Galphaq in Sheep Testis and Epididymis.

The reproductive function of G-protein subunit Galphaq (GNAQ), a member of the G protein alpha subunit family, has been extensively studied in humans and rats. However, no data is available on its status in ruminants. The objectives of this study were to evaluate the expression pattern of the GNAQ in the testis and epididymis of sheep by polymerase chain reaction (PCR). The mRNA expression levels were detected by real-time fluorescent quantitative PCR, and cellular localization of GNAQ in the testis and epididymis was examined by immunohistochemistry. Additionally, GNAQ protein was qualitatively evaluated via western blot, with the results indicating that similarities between GNAQ mRNA levels from sheep was highly conserved with those observed in Bos taurus and Sus scrofa. Our results also indicated that GNAQ exists in the caput and cauda epididymis of sheep, while GNAQ in the testis and epididymis was localized to Leydig cells, spermatogonial stem cells, spermatocytes, Sertoli cells, spermatid, principal cells, and epididymis interstitial cells. The concentrations of GNAQ mRNA and protein in the caput and cauda epididymis were significantly greater than those observed in the corpus epididymis (p<0.01) and testis (p<0.05). Our results indicated that GNAQ exists at high concentrations in the caput and cauda epididymis of sheep, suggesting that GNAQ may play an important role in gonad development and sperm maturation.

Significance of preoperative planning simulator for junior surgeons' training of pedicle screw insertion.

STUDY DESIGN: A prospective study to access the significance of preoperative planning simulator for junior surgeons' training of pedicle screw insertion. SUMMARY OF BACKGROUND DATA: Pedicle screw insertion is particularly challenging to carry out on patients with abnormal spine morphology, especially for the doctors who lack experience. Currently, preoperative planning for pedicle screw insertion is carried out using patient computed tomography and magnetic resonance imaging scans. In addition, there is no projection fluoroscopy provided to the user. OBJECTIVE: The aim of this study was to investigate the feasibility and efficacy of a 3-dimensional, patient-specific volume rendering combined with the projection fluoroscopy simulator for training junior surgeons with no experience of pedicle screw insertion, and to help identify the role such simulation has in surgical education. METHODS: Two junior surgeons with no experience of pedicle screw insertion were trained on the technique through the preoperative planning simulator; the operative time and the position of the pedicle screws were assessed before training (control group 1) and after training (experimental group) and compared with 2 senior spine surgeons with >10 years' experience of pedicle screw insertion (control group 2). RESULTS: The time of per pedicle screw insertion was 43.5±3.9 seconds in control group 1, 31.6±2.9 seconds in control group 2, and 50.8±3.7 seconds in experimental group. The relative position of the screw to the pedicle was graded regarding pedicle breach (I, no breach; II, <2 mm; III, 2-4 mm; IV, >4 mm). The pedicle breach grading I and II was 20 pedicle screws (20/56, 35.7%) in control group 1, 54 pedicle screws (54/56, 96.4%) in control group 2, and 44 pedicle screws (44/56, 78.6%) in the experimental group. There were significant differences between control group 1 and experimental group in the time of per pedicle screw insertion (P<0.001) and the rate of pedicle breach grading I and II (P<0.001). There were significant differences between control group 2 and experimental group in the time of per pedicle screw insertion (P<0.001) and the rate of pedicle breach grading I and II (P=0.004). CONCLUSIONS: The simulator offers many helpful features to the surgeon with respect to the surgical trainee learning the basic technique of pedicle screw insertion, using free-hand technique or under the guiding of intraoperative fluoroscopy. The surgical skills of the junior surgeons can be significantly improved through the training of simulator.

Photo-Controllable Smart Hydrogels for Biomedical Application: A Review.

Nowadays, smart hydrogels are being widely studied by researchers because of their advantages such as simple preparation, stable performance, response to external stimuli, and easy control of response behavior. Photo-controllable smart hydrogels (PCHs) are a class of responsive hydrogels whose physical and chemical properties can be changed when stimulated by light at specific wavelengths. Since the light source is safe, clean, simple to operate, and easy to control, PCHs have broad application prospects in the biomedical field. Therefore, this review timely summarizes the latest progress in the PCHs field, with an emphasis on the design principles of typical PCHs and their multiple biomedical applications in tissue regeneration, tumor therapy, antibacterial therapy, diseases diagnosis and monitoring, etc. Meanwhile, the challenges and perspectives of widespread practical implementation of PCHs are presented in biomedical applications. This study hopes that PCHs will flourish in the biomedical field and this review will provide useful information for interested researchers.

MF-Net: multi-scale feature extraction-integration network for unsupervised deformable registration.

Deformable registration plays a fundamental and crucial role in scenarios such as surgical navigation and image-assisted analysis. While deformable registration methods based on unsupervised learning have shown remarkable success in predicting displacement fields with high accuracy, many existing registration networks are limited by the lack of multi-scale analysis, restricting comprehensive utilization of global and local features in the images. To address this limitation, we propose a novel registration network called multi-scale feature extraction-integration network (MF-Net). First, we propose a multiscale analysis strategy that enables the model to capture global and local semantic information in the image, thus facilitating accurate texture and detail registration. Additionally, we introduce grouped gated inception block (GI-Block) as the basic unit of the feature extractor, enabling the feature extractor to selectively extract quantitative features from images at various resolutions. Comparative experiments demonstrate the superior accuracy of our approach over existing methods.