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Abstract This research explores the driving pathways for building a high-level rural e-commerce entrepreneurial ecosystem, providing valuable insights for responding to global policies promoting rural entrepreneurship and innovation and accelerating rural industrial development. However, existing studies on the “rural e-commerce entrepreneurial ecosystem” lack in-depth exploration. Based on the CAS (Complex Adaptive System) theory and TOE (Technological, Organizational, and Environmental) framework, we conducted a comprehensive case study across 31 Chinese provinces, focusing on seven key indicators within three dimensions: technology, organization, and environment. Employing the Fuzzy Set Qualitative Comparative Analysis (fsQCA) method, we examined key factors and driving pathways influencing the establishment of successful rural e-commerce entrepreneurial ecosystems in China. The main findings of this study are as follows: (1) The construction of a rural e-commerce entrepreneurial ecosystem is not driven by a single element but is the result of the combined effects of multiple facets and factors. (2) The number of leading agricultural enterprises and the establishment of e-commerce cooperatives are critical factors promoting the high-level construction of rural e-commerce entrepreneurial ecosystems. (3) Typical driving pathways for advancing the construction of rural e-commerce entrepreneurial ecosystems can be categorized into three types: technology-organization-government-driven, organization-environment-driven, and holistic synergy-driven. The research findings are instrumental in guiding the government to formulate systematic policies that support the development of leading agricultural enterprises and e-commerce cooperatives. Additionally, these policies should be tailored to the specific characteristics of different regions to create differentiated driving paths. Implementing these measures can effectively promote rural e-commerce entrepreneurship.

Abstract The purpose of this study is to investigate the influencing factors of abnormal pulmonary ventilation function in occupational exposed populations and to establish a risk prediction model. The findings will provide a basis for formulating corresponding strategies for the prevention and treatment of occupational diseases. The study focused on workers who underwent occupational health examinations in the year 2020. Statistical analysis was conducted using methods such as t-tests, chi-square tests, and multiple logistic regression analysis. Additionally, machine learning methods were employed to establish multiple models to address classification problems. Among the 7472 workers who participated in the occupational health examination, 1681 cases of abnormal pulmonary ventilation function were detected, resulting in a detection rate of 22.6%. Based on the analysis of occupational hazard data, a risk prediction model was established. Age, work tenure, type of the employing enterprise, and type of dust exposure are all identified as driving factors for abnormal pulmonary function. These factors were used as predictive variables for establishing the risk prediction model. Among the various models evaluated, the logistic regression model was found to be the optimal model for predicting abnormal pulmonary ventilation function.

Abstract Objective This study aimed to describe the clinical and genetic characteristics of Chinese patients with congenital fibrosis of the extraocular muscles (CFEOM), and to evaluate the phenotype–genotype correlations in these patients. Methods This was a retrospective study. Patients with CFEOM underwent detailed ophthalmic examinations and magnetic resonance imaging (MRI). Panel-based next-generation sequencing was performed to identify pathogenic variants of disease-causing genes. Results Sixty-two patients with CFEOM were recruited into this study. Thirty-nine patients were diagnosed with CFEOM1 and 23 with CFEOM3. Forty-nine of the 62 patients with CFEOM carried either KIF21A (41/49) or TUBB3 variants (8/49). Six known missense variants in the KIF21A and TUBB3 genes, and a novel variant (c.3906T > A, p.D1302E) in the KIF21A gene were detected. Most patients with CFEOM1 carrying the KIF21A mutation displayed isolated CFEOM, whereas patients with CFEOM3 carrying the TUBB3 mutation had a wide range of clinical manifestations, either CFEOM alone or syndromes. Nystagmus was also present in 12 patients with CFEOM. Furthermore, the MRI findings varied, ranging from attenuation of the extraocular muscles to dysgenesis of the cranial nerves and brain structure. Conclusions The novel variants identified in this study will further expand the spectrum of pathogenic variants in CFEOM-related genes. However, no phenotype–genotype correlations were established because of the diversity of the clinical characteristics of these patients.

Abstract Background The limited regenerative capacity of damaged neurons in adult mammals severely restricts neural repair. Although stem cell transplantation is promising, its clinical application remains challenging. Direct reprogramming, which utilizes cell plasticity to regenerate neurons, is an emerging alternative approach. Methods We utilized primary postnatal cortical astrocytes for reprogramming induced neurons (iNs) through the viral-mediated overexpression of the transcription factors Ngn2 and Pax6 (NP). Fluorescence-activated cell sorting (FACS) was used to enrich successfully transfected cells, followed by single-cell RNA sequencing (scRNA-seq) using the 10 × Genomics platform for comprehensive transcriptomic analysis. Results The scRNA-seq revealed that NP overexpression led to the differentiation of astrocytes into iNs, with percentages of 36% and 39.3% on days 4 and 7 posttransduction, respectively. CytoTRACE predicted the developmental sequence, identifying astrocytes as the reprogramming starting point. Trajectory analysis depicted the dynamic changes in gene expression during the astrocyte-to-iN transition. Conclusions This study elucidates the molecular dynamics underlying astrocyte reprogramming into iNs, revealing key genes and pathways involved in this process. Our research contributes novel insights into the molecular mechanisms of NP-mediated reprogramming, suggesting avenues for optimizing the efficiency of the reprogramming process.

Abstract Objective To investigate the characteristics of eye movement in children with anisometropic amblyopia, and to compare those characteristics with eye movement in a control group. Methods 31 children in the anisometropic amblyopia group (31 amblyopic eyes in group A, 31 contralateral eyes in group B) and 24 children in the control group (48 eyes in group C). Group A was subdivided into groups Aa (severe amblyopia) and Ab (mild-moderate amblyopia). The overall age range was 6–12 years (mean, 7.83 ± 1.79 years). All children underwent ophthalmic examinations; eye movement parameters including saccade latency and amplitude were evaluated using an Eyelink1000 eye tracker. Data Viewer and MATLAB software were used for data analysis. Results Mean and maximum saccade latencies, as well as mean and maximum saccade amplitudes, were significantly greater in group A than in groups B and C before and after treatment (P

Klebsiella pneumoniae is a common, conditionally pathogenic bacterium that often has a multidrug-resistant phenotype, leading to failure of antibiotic therapies. It can therefore induce serious diseases, including community-acquired pneumonia and bloodstream infections. As an emerging alternative to antibiotics, phages are considered key to solving the problem of drug-resistant bacterial infections. Here, we report a novel phage, pK3–24, that mainly targets ST447 K. pneumoniae. Phage pK3–24 is a T7-like short-tailed phage with a fast adsorption capacity that forms translucent plaques with halos on bacterial lawns. The optimal multiplicity of infection (MOI) is 0.01, and the average burst size is 50 PFU/mL. Phage pK3–24 shows environmental stability, surviving at below 50 °C and at pH values of 6–10. It has a double-stranded DNA genome of 40,327 bp and carries no antibiotic-resistance, virulence, or lysogeny genes. Phylogenetic analysis assigned phage pK3–24 to the genus Przondovirus as a new species. Phage pK3–24 inhibited the production of biofilm. Moreover, treatment with pK3–24 at doses with an MOI > 1 effectively reduced the mortality of Galleria mellonella larvae infected with ST447 K. pneumoniae.

The purpose of this study was to investigate the relationship between childhood chronic stress(CCS), Protein kinase C beta (PRKCB) methylation and adolescent major depressive disorder (MDD). After recruiting 100 adolescents with MDD and 50 healthy controls (HCs), we evaluated the severity of CCS. PRKCB methylation was assessed by pyrosequencing using whole blood-derived DNA. To explore the relationship between CCS, PRKCB and adolescent MDD, we conducted correlation analysis and regression analysis, and constructed multiplicative interaction models and generalized linear models. PRKCB methylation and CCS were both found to be associated with MDD, and CCS was associated with PRKCB methylation. No significant CCS-PRKCB methylation interactions were observed. However, we found the interaction of CCS and MDD on PRKCB methylation. Our results found that PRKCB methylation was influenced by CCS and the disease itself, and PRKCB methylation was significantly positively associated with MDD severity, suggesting that PRKCB methylation may be a potential biomarker for adolescent MDD. This study is a cross-sectional observational study, which cannot draw the conclusion of causality. Prospective cohort studies are needed to further examine the relationship between CCS, adolescent MDD, and PRKCB methylation.

The quality of silage is uneven, which is due to the difference of epiphytic microorganisms of raw materials. To improve the quality of fermentation, the quantity and composition of epiphytic microorganisms are usually analyzed to better prepare silage. In this research, plate coating method and 16S high-throughput sequencing were used to analyze the differences in the quantity and composition of rice epiphytic microorganisms during different growth stages. The Lactic acid bacteria (LAB) and aerobic bacteria were the highest at the flowering stage, the yeast was the highest at the milk ripening stage, and the mould was the highest at the full ripening stage. And the growth stage also had a great influence on the composition of epiphytic bacterial community, at the phylum level, it was mainly composed of Proteobacteria. And at the genus level, Pantoea, Acinetobacter, Pseudomonas and Chryseobacterium were dominanted at the flowering stage; Pantoea, Stenotrophomonas and Sphingobacterium were dominanted at the milk ripening stage; Acinetobacter, Pantoea, Chryseobacterium and Lactococcus were dominanted at the dough stage; Acinetobacter and Klebsiella were dominated at the full ripening stage. Overall, the growth stage significantly affected the quantity and composition of rice epiphytic microorganisms. Therefore, rice silage can be modulated reasonably according to the number and composition of epiphytic microorganisms in different growth stages.

Long-term and/or excessive ethanol intake can lead to alcoholic liver disease (ALD). Gut dysbiosis contributes to a critical role in the pathogenesis of ALD. Here, we explored the impact of viable and dead Lacticaseibacillus rhamnosus CCFM1060 on the gut microbiota of alcohol-treated mice. The findings indicated that CCFM1060 V and D (viable and dead) administration improved the gut microbiota composition. Specifically, CCFM1060 D restored the abundance of butyric acid-producing bacteria, Alistipes, Lachnospiraceae, and Ruminococcaceae in ethanol-induced mice; CCFM1060 V improved the abundance of g_Dubsiela, g_Bifdobacterium, g_Ruminococcaceae_UCG_014_2, the SCFA-producing bacteria which were decreased in alcoholic hepatitis. Furthermore, the interventions improved the microstructure of the ileum, including the villi and crypts. They reinforced the intestinal barrier by increasing the RNA and protein expression of claudin-1, occludin, and ZO-1 in the colon. Also, they have been shown to enhance the liver’s antioxidant function by suppressing the synthesis of MDA and boosting the concentrations of GSH and SOD through the Nrf2/HO-1 pathway. In addition, interventions can suppress mice’s levels of TNF-α and IL-6, IL-1β in serum, presumably via the TRL4/MyD88/NF-κB pathway. Together, these results suggest that CCFM1060 can modulate the gut microbiota structure, reinforce the intestinal barrier, improve intestinal homeostasis, enhance liver antioxidant capacity, and mitigate inflammation response, eventually protecting the liver from ethanol-induced injury.

Abstract The asymmetric calibration method is a very interesting technology for differential output probes. It has been proven that the asymmetric calibration method (ACM) can broaden the application scenarios of the electromagnetic field dual probe. However, ACM is not verified on a back‐to‐back double‐loop differential magnetic field probing system. This paper calibrates a back‐to‐back double‐loop differential magnetic field probing system by inserting a connector for creating an asymmetry using ACM. The calibration results show that ACM can be used to calibrate an asymmetric back‐to‐back double‐loop differential magnetic field probing system. The calibration results are further verified by measuring the standing wave magnetic field, and the verification results show that the asymmetric calibration method is effective in eliminating the asymmetry of the back‐to‐back double‐loop differential magnetic field probing system and the work frequency band reaches up to 12 GHz.

In the ever-changing sector of education, the use of technology has become critical to innovation and increased learning opportunities. This research illustrates the intricate connections that exist between augmented reality [AR] and tremendous educational attributes, thinking about how AR might modify conventional teaching approaches. The important motive is to offer an entire knowledge of how augmented fact influences crucial areas of the educational revel in. Utilizing a quantitative studies method, the study seems at how AR impacts pupil behavior, creativity, and instructional practices, in addition to the mediating and moderating elements that influence these relationships. An online questionnaire was used to collect data from 444 random samples. The relationships between AR, teacher competence, technology acceptance, and educational outcomes were examined. Data shows that augmented reality has a positive effect on creativity, student behavior, and teaching strategies. Teacher competence moderated these relationships, highlighting the critical role of teachers in successfully integrating augmented reality, while technology acceptance emerged as a significant mediator demonstrating the significance of user perceptions. This study has critical implications for educators, administrators, and policymakers interested in using AR to improve instructional development, as well as for the field of generation-infused schooling administration. Additionally, it advances theoretical frameworks through a better understanding of the complicated tactics that impact the uptake and acceptability of AR.

Abstract Epidermal growth factor receptor (EGFR) exon 19 deletion is a major driver for the drug resistance of non-small cell lung cancer (NSCLC). Identification small inhibitor capable of selectively inhibiting EGFR-19del NSCLC is a desirable strategy to overcome drug resistance in NSCLC. This study aims to screen an inhibitor for EGFR exon 19 deletion cells and explore its underlying mechanism. High through-put screen was conducted to identify an inhibitor for EGFR-19del NSCLC cells. And tenovin-3 was identified as a selective inhibitor of PC9 cells, an EGFR-19del NSCLC cells. Tenovin-3 showed particular inhibition effect on PC9 cells proliferation through inducing apoptosis and ferroptosis. Mechanistically, tenovin-3 might induce the apoptosis and ferroptosis of PC9 cells through mitochondrial pathway, as indicated by the change of VDAC1 and cytochrome c (cyt c). And bioinformatics analyses showed that the expression levels of SLC7A11 and CPX4 were correlated with NSCLC patient’s survival. Our findings provide evidences for tenovin-3 to be developed into a novel candidate agent for NSCLC with EGFR exon 19 deletion. Our study also suggests that inducing ferroptosis may be a therapeutic strategy for NSCLC with EGFR exon 19 deletion.

In the treatment of central nervous system disease, the blood–brain barrier (BBB) is a major obstruction to drug delivery that must be overcome. In this study, we propose a brain-targeted delivery strategy based on selective opening of the BBB. This strategy allows some simple bare nanoparticles to enter the brain when mixed with special opening material; however, the BBB still maintains the ability to completely block molecules from passing through. Based on the screening of BBB opening and matrix delivery materials, we determined that phospholipase A2-catalyzed 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine liposomes can efficiently carry drugs into the brain immediately. At an effective dose, this delivery system is safe, especially with its effect on the BBB being reversible. This mix & act delivery system has a simple structure and rapid preparation, making it a strong potential candidate for drug delivery across the BBB.

Objective: Ischemic stroke (IS) is a significant health concern with high disability and fatality rates despite available treatments. Immune cells and cuproptosis are associated with the onset and progression of IS. Investigating the interaction between cuproptosis-related genes (CURGs) and immune cells in IS can provide a theoretical basis for IS treatment. Methods: We obtained IS datasets from the Gene Expression Omnibus (GEO) and employed machine learning to identify CURGs. The diagnostic efficiency of the CURGs was evaluated using receiver operating characteristic (ROC) curves. KEGG and gene set enrichment analysis (GSEA) were also conducted to identify biologically relevant pathways associated with CURGs in IS patients. Single-cell analysis was used to confirm the expression of 19 CURGs, and pathway activity calculations were performed using the AUCell package. Additionally, a risk prediction model for IS patients was developed, and core modules and hub genes related to IS were identified using weighted gene coexpression network analysis (WGCNA). We classified IS patients using a method of consensus clustering. Results: We established a precise diagnostic model for IS. Enrichment analysis revealed major pathways, including oxidative phosphorylation, the NF-kappa B signaling pathway, the apoptosis pathway, and the Wnt signaling pathway. At the single-cell level, compared to those in non-IS samples, 19 CURGs were primarily overexpressed in the immune cells of IS samples and exhibited high activity in natural killer cell-mediated cytotoxicity, steroid hormone biosynthesis, and oxidative phosphorylation. Two clusters were obtained through consensus clustering. Notably, immune cell types including B cells, plasma cells, and resting NK cells, varied between the two clusters. Furthermore, the red module and hub genes associated with IS were uncovered. The expression patterns of CURGs varied over time. Conclusion: This study developed a precise diagnostic model for IS by identifying CURGs and evaluating their interaction with immune cells. Enrichment analyses revealed key pathways involved in IS, and single-cell analysis confirmed CURG overexpression in immune cells. A risk prediction model and core modules associated with IS were also identified.

We collected respiratory specimens from 128 pediatric patients diagnosed with pneumonia in Beijing in late 2023. Mycoplasma pneumoniae was detected in 77.3% (99/128) patients, with 36.4% (4/11), 82.9% (34/41), 80.3% (61/76) in children aged less than 3 years, 3–6 years, over 7 years, respectively. Mycoplasma pneumoniae (M. pneumoniae) was characterized using P1 gene typing, MLVA typing and sequencing of domain V of the 23S rRNA gene. P1 gene type 1 (P1-1; 76.1%, 54/71) and MLVA type 4-5-7-2 (73.7%, 73/99) were predominant. MLVA identified a new genotype: 3–4–6-2. Macrolide resistance-associated mutations were detected in 100% of samples, with A2063G accounting for 99% and A2064G for 1%. The positive rate of M. pneumoniae was higher compared to previous reports, especially in children less than 3 years, suggesting a M. pneumoniae epidemic showing a younger age trend occurred in late 2023 in Beijing, China. Higher proportions of macrolide-resistant M. pneumoniae, P1-1 and 4-5-7-2 genotype M. pneumoniae indicated increased macrolide resistance rate and genotyping shift phenomenon, which might be attributable to this epidemic. Additionally, complete clinical information from 73 M. pneumoniae pneumonia inpatients were analyzed. The incidence of severe M. pneumoniae pneumonia was 56.2% (41/73). Mycoplasma pneumoniae pneumonia patients exhibited longer duration of fever, with a median value of 10.0 days (IQR, 8.0–13.0), and higher incidence of complications (74.0%, 54/73). However, in this cohort, we found that the severity of M. pneumoniae pneumonia, co-infection, or complications were not associated with M. pneumoniae P1 gene or MLVA types. Clinicians should be aware that patients infected with macrolide-resistant M. pneumoniae exhibited more severe clinical presentations.

Abstract Background High‐frequency repeated transcranial magnetic stimulation (rTMS) stimulating the primary motor cortex (M1) is an alternative, adjunctive therapy for improving the motor symptoms of Parkinson's disease (PD). However, whether the high frequency of rTMS positively correlates to the improvement of motor symptoms of PD is still undecided. By controlling for other parameters, a disease animal model may be useful to compare the neuroprotective effects of different high frequencies of rTMS. Objective The current exploratory study was designed to compare the protective effects of four common high frequencies of rTMS (5, 10, 15, and 20 Hz) and iTBS (a special form of high‐frequency rTMS) and explore the optimal high‐frequency rTMS on an animal PD model. Methods Following high frequencies of rTMS application (twice a week for 5 weeks) in a MPTP/probenecid‐induced chronic PD model, the effects of the five protocols on motor behavior as well as dopaminergic neuron degeneration levels were identified. The underlying molecular mechanisms were further explored. Results We found that all the high frequencies of rTMS had protective effects on the motor functions of PD models to varying degrees. Among them, the 10, 15, and 20 Hz rTMS interventions induced comparable preservation of motor function through the protection of nigrostriatal dopamine neurons. The enhancement of brain‐derived neurotrophic factor (BDNF), dopamine transporter (DAT), and vesicular monoamine transporter 2 (VMAT‐2) and the suppression of TNF‐α and IL‐1β in the nigrostriatum were involved in the process. The efficacy of iTBS was inferior to that of the above three protocols. The effect of 5 Hz rTMS protocol was weakest. Conclusions Combined with the results of the present study and the possible side effects induced by rTMS, we concluded that 10 Hz might be the optimal stimulation frequency for preserving the motor functions of PD models using rTMS treatment.

Abstract Background Tumor angiogenesis inhibitors have been applied for non-small cell lung cancer (NSCLC) therapy. However, the drug resistance hinders their further development. Intercellular crosstalk between lung cancer cells and vascular cells was crucial for anti-angiogenenic resistance (AAD). However, the understanding of this crosstalk is still rudimentary. Our previous study showed that Glioma-associated oncogene 1 (Gli1) is a driver of NSCLC metastasis, but its role in lung cancer cell-vascular cell crosstalk remains unclear. Methods Conditioned medium (CM) from Gli1-overexpressing or Gli1-knockdown NSCLC cells was used to educate endothelia cells and pericytes, and the effects of these media on angiogenesis and the maturation of new blood vessels were evaluated via wound healing assays, Transwell migration and invasion assays, tube formation assays and 3D coculture assays. The xenograft model was conducted to establish the effect of Gli1 on tumor angiogenesis and growth. Angiogenic antibody microarray analysis, ELISA, luciferase reporte, chromatin immunoprecipitation (ChIP), bFGF protein stability and ubiquitination assay were performed to explore how Gli1 regulate bFGF expression. Results Gli1 overexpression in NSCLC cells enhanced the endothelial cell and pericyte motility required for angiogenesis required for angiogenesis. However, Gli1 knockout in NSCLC cells had opposite effect on this process. bFGF was critical for the enhancement effect on tumor angiogenesis. bFGF treatment reversed the Gli1 knockdown-mediated inhibition of angiogenesis. Mechanistically, Gli1 increased the bFGF protein level by promoting bFGF transcriptional activity and protein stability. Importantly, suppressing Gli1 with GANT-61 obviously inhibited angiogenesis. Conclusion The Gli1-bFGF axis is crucial for the crosstalk between lung cancer cells and vascular cells. Targeting Gli1 is a potential therapeutic approach for NSCLC angiogenesis.

Abstract Background Congenital stationary night blindness (CSNB) is an inherited retinal disorder. Most of patients have myopia. This study aims to describe the clinical and genetic characteristics of fifty-nine patients with CSNB and investigate myopic progression under genetic cause. Results Sixty-five variants were detected in the 59 CSNB patients, including 32 novel and 33 reported variants. The most frequently involved genes were NYX, CACNA1F, and TRPM1. Myopia (96.61%, 57/59) was the most common clinical finding, followed by nystagmus (62.71%, 37/59), strabismus (52.54%, 31/59), and nyctalopia (49.15%, 29/59). An average SE of -7.73 ± 3.37 D progressed to -9.14 ± 2.09 D in NYX patients with myopia, from − 2.24 ± 1.53 D to -4.42 ± 1.43 D in those with CACNA1F, and from − 5.21 ± 2.89 D to -9.24 ± 3.16 D in those with TRPM1 during the 3-year follow-up; the TRPM1 group showed the most rapid progression. Conclusions High myopia and strabismus are distinct clinical features of CSNB that are helpful for diagnosis. The novel variants identified in this study will further expand the knowledge of variants in CSNB and help explore the molecular mechanisms of CSNB.

Purpose: Mutations of G protein-coupled receptor 143 (GPR143) and FERM domain containing 7 (FRMD7) may result in congenital nystagmus (CN) in the first 6 months of life. We aimed to compare the differences in ocular oscillations between patients with these two gene mutations as well as the functional and structural changes in their retinas and visual pathways. Methods: Medical records were retrospectively reviewed to identify patients of congenital nystagmus with confirmed mutations in either GPR143 or FMRD7 genes from January 2018 to May 2023. The parameters of the ocular oscillations were recorded using Eyelink 1000 Plus. The retinal structure and function were evaluated using optical coherence tomography and multi-focal electroretinography (mERG). The visual pathway and optical nerve projection were evaluated using visual evoked potentials. The next-generation sequencing technique was used to identify the pathogenic variations in the disease-causing genes for CN. Results: Twenty nystagmus patients of GPR143 and 21 patients of FMRD7 who had been confirmed by molecular testing between January 2018 and May 2023 were included. Foveal hypoplasia was detected only in patients with the GPR143 pathogenic variant. mERG examination showed a flat response topography in the GPR143 group compared to the FRMD7 group. VEP showed that bilateral amplitude inconsistency was detected only in the patients with GPR143 gene mutation. The amplitude and frequency of the ocular oscillations were not found to differ between patients with two different genetic mutations. Conclusions: Although the etiology and molecular mechanisms are completely different between CN patients, they may have similar ocular oscillations. A careful clinical examination and electrophysiological test will be helpful in making a differential diagnosis. Our novel identified variants will further expand the spectrum of the GPR143 and FRMD7 variants.

Immune infiltration plays a pivotal role in the pathogenesis of ischemic stroke. A novel form of cell death known as disulfidptosis has emerged in recent studies. However, there is currently a lack of research investigating the regulatory mechanism of disulfidptosis-related genes in immune infiltration during ischemic stroke. Using machine learning methods, we identified candidate key disulfidptosis-related genes (DRGs). Subsequently, we performed an analysis of immune cell infiltration to investigate the dysregulation of immune cells in the context of ischemic stroke. We assessed their diagnostic value by employing receiver operating characteristic (ROC) curves. To gain further insights, we conducted functional enrichment analyses to elucidate the signaling pathways associated with these seven DRGs. We identified two distinct subclusters based on the expression patterns of these seven DRGs. The unique roles of these subclusters were further evaluated through KEGG analysis and immune infiltration studies. Furthermore, we validated the expression profiles of these seven DRGs using both single-cell datasets and external datasets. Lastly, molecular docking was performed to explore potential drugs for the treatment of ischemic stroke. We identified seven DRGs. The seven DRGs are related to immune cells. Additionally, these seven DRGs also demonstrate potential diagnostic value in ischemic stroke. Functional enrichment analysis highlighted pathways such as platelet aggregation and platelet activation. Two subclusters related to disulfidptosis were defined, and functional enrichment analysis of their differentially expressed genes (DEGs) primarily involved pathways like cytokine-cytokine receptor interaction. Single-cell analysis indicated that these seven DRGs were primarily distributed among immune cell types. Molecular docking results suggested that genistein might be a potential therapeutic drug. This study has opened up new avenues for exploring the causes of ischemic stroke and developing potential therapeutic targets.

Abstract Although VEGF-B was discovered as a VEGF-A homolog a long time ago, the angiogenic effect of VEGF-B remains poorly understood with limited and diverse findings from different groups. Notwithstanding, drugs that inhibit VEGF-B together with other VEGF family members are being used to treat patients with various neovascular diseases. It is therefore critical to have a better understanding of the angiogenic effect of VEGF-B and the underlying mechanisms. Using comprehensive in vitro and in vivo methods and models, we reveal here for the first time an unexpected and surprising function of VEGF-B as an endogenous inhibitor of angiogenesis by inhibiting the FGF2/FGFR1 pathway when the latter is abundantly expressed. Mechanistically, we unveil that VEGF-B binds to FGFR1, induces FGFR1/VEGFR1 complex formation, and suppresses FGF2-induced Erk activation, and inhibits FGF2-driven angiogenesis and tumor growth. Our work uncovers a previously unrecognized novel function of VEGF-B in tethering the FGF2/FGFR1 pathway. Given the anti-angiogenic nature of VEGF-B under conditions of high FGF2/FGFR1 levels, caution is warranted when modulating VEGF-B activity to treat neovascular diseases.

ABSTRACT Purpose: Chronic inflammation in the liver is a key trigger for liver injury and fibrosis in various liver diseases. Given the anti-inflammatory and antioxidant effects of Saffron, this study aimed to investigate the pharmacological effects of Saffron on hepatic inflammation and fibrosis. Methods: The mice model of hepatic fibrosis was constructed using CCl4, and Saffron was administered at low (10 mg/kg) and high (20 mg/kg) doses by gavage. Then, the changes in liver function, liver inflammation and fibrosis markers were evaluated. The effects and mechanisms of Saffron on hepatic stellate cells were further investigated in in-vitro experiments. Results: Saffron improved liver function, reduced liver inflammation and attenuated liver fibrosis in a dose-dependent manner in hepatic fibrosis mice. Furthermore, Western blotting showed that Saffron significantly inhibited JAK/STAT3 phosphorylation in fibrotic livers. Conclusions: Saffron can attenuate liver fibrosis by inhibiting the JAK/STAT3 pathway and the activation of hepatic stellate cell, providing a theoretical basis for the development of new anti-fibrotic drugs.

AbstractMessenger RNA (mRNA) has become one of the most potential drugs in recent years. However, efficient and safe delivery of fragile and easily degradable mRNA is a major challenge. Appropriate delivery system (DS) determines the final effect of mRNA. Cationic lipids play a crucial and decisive role in the entire DS, but also cause huge biosafety problems due to the high toxicity. In this study, a new DS for mRNA delivery that combines negatively charged phospholipids was developed in order to neutralize the positive charge and thus increase the safety. Further, the factors affecting mRNA transfection from cell to animal were investigated. The mRNA DS with optimum condition of lipid composition, proportions, structure, and transfection time was synthesized. Adding an appropriate amount of the anionic lipid to liposomes could increase the safety while maintaining the original transfection efficiency. For transporting mRNA in vivo, requirements regarding the mRNA encapsulation and releasing rate should be further considered to optimize DS design and preparation.

Abstract Background Head and Neck Squamous Cell Carcinoma is a malignant tumor with high morbidity and mortality. The MMP family plays an important role in tumor invasion and metastasis. However, the mechanistic value of the MMP family as a therapeutic target and prognostic biomarker in HNSC has not been fully elucidated. Methods Oncomine, UALCAN, GEPIA, cBioportal, GeneMANIA, STRING, DAVID6.8, TRRUST, TIMER and Linkedomics were used for analysis. Results The mRNA expression levels of MMP1, MMP3, ILF3, MMP7, MMP9, MMP10, MMP11, MMP12, MMP13 and MMP16 were higher in HNSC than those in normal tissues, while the mRNA expression level of MMP15 was reduced. The relative expression levels of MMP1 and MMP14 were the highest in HNSC tissues. A significant correlation was found between the expression of MMP3, MMP11, MMP25 and the pathological stage of HNSC patients. There was no significant associations between all the MMP family members expression levels and DFS. Increased mRNA levels of MMP1, MMP8 and MMP25 were significantly associated with OS. In addition, we investigated the genetic changes of the MMP family in HNSC and found that all the MMP family members had genetic changes, most of which were amplification and depth loss. In the analysis of neighbor gene network and protein interaction, we found that the MMP family interacted with 25 neighboring genes, except for ILF3, MMP19, MMP20, MMP21, MMP23B, MMP27 and MMP28, other MMP proteins interacted with each other. Functional enrichment analysis showed that the MMP family could be present in the extracellular matrix, regulate peptidase activity, and participate in the catabolism of collagen. Meanwhile, we identified the transcription factor targets and kinase targets of the MMP family and found that ATM and ATR were the two most common kinase targets in the MMP family. We also found a significant correlation between the MMP family expression and immune cell infiltration. Cox proportional risk model analysis showed that macrophages, MMP14, MMP16, and MMP19 were significantly associated with clinical outcomes in HNSC patients. Conclusion The MMP family might serve as therapeutic target and prognostic biomarker in HNSC.

Abstract Objective To investigate the role of standard patients (SPs) and examiners as assessors for scoring in the dental objective structured clinical examination (OSCE) system and to evaluate the scoring differences between them. Methods We developed the doctor-patient communication and clinical examination station in the OSCE system. The examination time of this station was 10 min, and the examination institution wrote the script and recruited SPs. A total of 146 examinees who received standardized resident training at the Nanjing Stomatological Hospital, Medical School of Nanjing University between 2018 and 2021 were assessed. They were scored by SPs and examiners according to the same scoring rubrics. Subsequently, the SPSS software was used to analyze the examination results of different assessors and evaluate the consistency. Results The average score of all examinees provided by SPs and examiners was 90.45 ± 3.52 and 91.53 ± 4.13, respectively. The consistency analysis showed that the intraclass correlation coefficient was 0.718, which was indicative of medium consistency. Conclusion Our findings showed that SPs could be used directly as assessors, as they could provide a simulated and realistic clinical setting and create favorable conditions for comprehensive competence training and improvement for medical students.

Abstract Background Sulfur (S) has been widely used to alleviate cadmium (Cd) toxicity and control Cd accumulation in rice under waterlogging conditions. However, the results are contradictory, and the reasons remain unclear. This could be because most studies rarely simultaneously monitor the processes of S-induced soil Cd bioavailability and Cd accumulation in rice throughout its growth period. A pot experiment was conducted to investigate the influence of two sulfur levels (0, and 30 mg S kg−1) on Cd concentration and translocation in rice at three growth stages (booting, filling and maturity) under waterlogging conditions. Paddy soil deficient in S but contaminated with Cd (10.16 mg Cd kg−1) was used for the pot experiment. Results S application increased concentrations of Cd in grain at the filling stage partially because S induced the promotion of Cd transfer from roots to stems, leaves, and grains, and S induced the accumulation and fixation of Cd in iron plaques at the filling stage. However, the application of S significantly reduced Cd concentrations in brown rice at the maturity stage, which could be attributed to three aspects, as described below. First, S supply reduced the availability of Cd in soil and iron plaque on the root surface by reducing dissolved Cd in soil pore water and transferring Cd from iron plaque on the root surface to roots. Second, S supply inhibited the transfer of Cd in other tissues to brown rice based on Cd transfer factors from roots, stems, leaves, and husks to brown rice, which were obviously lower with S supply than without S supply at the maturity stage. Third, S induced the dilution of Cd in brown rice because the application of S significantly increased brown rice biomass by 215%. Conclusions A S-induced decline in Cd accumulation in brown rice was related to S-regulated Cd transfer among rice plants, S-induced promotion of rice growth and a decrease in Cd bioavailability in S-deficient but Cd-contaminated paddy soil under waterlogging conditions. This study provides valuable information for growing rice in low-S and Cd-contaminated paddy soil and reducing the risk of Cd in rice to humans. Graphical abstract

ABSTRACT Background: This study aimed to explore the prevalence and clinical risk factors in patients diagnosed with incidental prostate cancer (IPC) during certain surgeries (transurethral resection of the prostate [TURP], open prostatectomy [OP], and holmium laser enucleation of the prostate [HoLEP]) after clinically suspected benign prostatic hyperplasia (BPH). Materials and Methods: Literature search of the MEDILINE, Web of Science, Embase, and Cochrane Library databases was performed to identify eligible studies published before June 2021. Multivariate adjusted odds ratios (ORs) and associated 95% confidence intervals (CIs) of the prevalence and clinical risk factors of IPC were calculated using random or fixed-effect models. Results: Twenty-three studies were included in the meta-analysis. Amongst the 94.783 patients, IPC was detected in 24.715 (26.1%). Results showed that the chance of IPC detection (10%, 95% CI: 0.07-4.00; P

Metastasis is crucial for the mortality of non-small cell lung carcinoma (NSCLC) patients. The epithelial–mesenchymal transition (EMT) plays a critical role in regulating tumor metastasis. Glioma-associated oncogene 1 (Gli1) is aberrantly active in a series of tumor tissues. However, the molecular regulatory relationships between Gli1 and NSCLC metastasis have not yet been identified. Herein, we reported Gli1 promoted NSCLC metastasis. High Gli1 expression was associated with poor survival of NSCLC patients. Ectopic expression of Gli1 in low metastatic A549 and NCI-H460 cells enhanced their migration, invasion abilities and facilitated EMT process, whereas knock-down of Gli1 in high metastatic NCI-H1299 and NCI-H1703 cells showed an opposite effect. Notably, Gli1 overexpression accelerated the lung and liver metastasis of NSCLC in the intravenously injected metastasis model. Further research showed that Gli1 positively regulated Snail expression by binding to its promoter and enhancing its protein stability, thereby facilitating the migration, invasion and EMT of NSCLC. In addition, administration of GANT-61, a Gli1 inhibitor, obviously suppressed the metastasis of NSCLC. Collectively, our study reveals that Gli1 is a critical regulator for NSCLC metastasis and suggests that targeting Gli1 is a prospective therapy strategy for metastatic NSCLC.

Abstract Background Growing evidences have indicated neurodevelopmental disorders in infantile esotropia (IE). However, few studies have analyzed the characteristics of large‐scale functional networks of IE patients or their postoperative network‐level alterations. Methods Here, individuals with IE (n = 32) and healthy subjects (n = 30) accomplished the baseline clinical examinations and resting‐state MRI scans. A total of 17 IE patients also underwent corrective surgeries and completed the longitudinal clinical assessments and resting‐state MRI scans. Linear mixed effects models were applied for cross‐sectional and longitudinal network‐level analyses. Correlation analysis was performed to assess the relationship between longitudinal functional connectivity (FC) alterations and baseline clinical variables. Results In cross‐sectional analyses, network‐level FC were apparently aberrant in IE patients compared to controls. In longitudinal analyses, intra‐ and internetwork connectivity were observed with significant alterations in postoperative IE patients compared to the preoperative counterparts. Longitudinal FC changes are negatively correlated to the age at surgery in IE. Conclusions Obviously, altered network‐level FC benefiting from the corrective surgery serves as the neurobiological substrate of the observed improvement of stereovision, visuomotor coordination, and emotional management in postoperative IE patients. Corrective surgery should be performed as early as possible to obtain more benefits for IE in brain function recovery.

BackgroundNon-suicidal self-injury (NSSI) is a major public health concern among adolescents. Further research is needed into contributors to this behavior, in particular among adolescents with psychiatric disorders. The aim of the present study was to explore the impact of life events and emotional stress on NSSI among hospitalized psychiatric adolescents.MethodsIn this cross-sectional study, 505 Chinese psychiatric adolescent inpatients 10–19 years old completed questionnaires about sociodemographic characteristics and NSSI as well as the Adolescent Self-Rating Life Events Checklist (ASLEC), the State–Trait Anxiety Inventory Form Y, and the Center for Epidemiological Studies Depression Scale. Chi-square test was used to compare the incidence of NSSI in psychiatric adolescent patients with different sociodemographic. T-test was used to compare the total scores and dimension scores of the ASLEC, STAI-Y, and CES-D between the NSSI group and the non-NSSI group. A binary logistic regression model was built to explore the relationships among sociodemographic characteristics, questionnaire scores and NSSI.ResultsMost psychiatric adolescent inpatients (393, 77.8%) reported NSSI behavior. The higher risk for NSSI was observed among female (odds ratio [OR] 2.665, 95% confidence interval [CI] 1.575–4.510), younger adolescents (10–14 years; OR 2.021, 95% CI 1.258–3.245), with a suicide history (OR 2.479, 95% CI 1.549–3.967), or with depression symptom (OR 3.217, 95% CI 1.572–6.582) and those with higher scores of ASLEC (OR 1.019, 95% CI 1.010–1.029).ConclusionOur study in China is one of the first to apply to adolescent inpatients the diagnostic criteria of NSSI in the latest edition of the Diagnostic and Statistical Manual of Mental Disorders. Our analysis suggests that NSSI prevalence is disturbingly high among adolescents with mental illness in China. A better understanding of contributing factors, especially negative life events and negative emotions, may guide interventions that can reduce its prevalence.

The ketogenic diet (KD) has shown excellent performance in the treatment of refractory epilepsy, but how it works is not yet fully understood. Gut microbiota is associated with various neurological disorders through the brain-gut axis. Different dietary patterns have different effects on the composition and function of gut microbiota. Here, by analyzing fecal samples from some patients with mitochondrial epilepsy before and after KD treatment through 16SrRNA sequencing, we found that KD intervention reduced the abundance of Firmicutes in the patient’s gut, while the abundance of Bacteroidota increased in the KD group. LefSe analysis showed that Actinobacteriota, Phascolarctobacterium had significant advantages in the control group, while Bacteroides increased significantly after KD intervention, especially Bacteroides fragilis. Functional analysis showed that there were significant differences in 12 pathways in level 3. These changes suggest that KD can change the composition and diversity of the gut microbiota in patients and affect their function. Changes in specific bacterial groups in the gut may serve as biomarkers for the therapeutic effects of KD on epilepsy.

Abstract Objective We aimed to investigate the effect of differentially methylated genes and chronic childhood stress on the development of depressive symptoms in Chinese adolescents, as well as to test whether methylation at baseline can be used as a predictor of remission at follow-up after six weeks of treatment. Methods After recruiting 87 MDD patients and 53 healthy controls, we compared demographic and baseline clinical characteristics. The Childhood Chronic Stress Questionnaire was used to assess stress caused by early-life events. MDD patients underwent six weeks of treatment, and response to treatment was assessed using the Beck Depression Inventory-II. In addition, four MDD patients and five controls were randomly chosen for genome-wide methylation analysis. Results The gene RPS6KA5 showed significant methylation differences between the two groups. Severity of chronic childhood stress was significantly associated with increased risk of depression in adolescents, but not with treatment response. Baseline RPS6KA5 methylation can predict remission after six weeks of treatment. We did not observe any interaction between RPS6KA5 methylation and chronic childhood stress. Conclusions Our results suggest that RPS6KA5 methylation can be used as a predictor of response to treatment in adolescent MDD patients. Here we offer new evidence for the role of epigenetics in early response to treatment of depression. Trial registration ChiCTR, ChiCTR2000033402, 31/05/2020, http://www.chictr.org.cn/index.aspx

Xenobiotics can easily harm human lungs owing to the openness of the respiratory system. Identifying pulmonary toxicity remains challenging owing to several reasons: 1) no biomarkers for pulmonary toxicity are available that might help to detect lung injury; 2) traditional animal experiments are time-consuming; 3) traditional detection methods solely focus on poisoning accidents; 4) analytical chemistry methods hardly achieve universal detection. An in vitro testing system able to identify the pulmonary toxicity of contaminants from food, the environment, and drugs is urgently needed. Compounds are virtually infinite, whereas toxicological mechanisms are countable. Therefore, universal methods to identify and predict the risks of contaminants can be designed based on these well-known toxicity mechanisms. In this study, we established a dataset based on transcriptome sequencing of A549 cells upon treatment with different compounds. The representativeness of our dataset was analyzed using bioinformatics methods. Artificial intelligence methods, namely partial least squares discriminant analysis (PLS-DA) models, were employed for toxicity prediction and toxicant identification. The developed model predicted the pulmonary toxicity of compounds with a 92 % accuracy. These models were submitted to an external validation using highly heterogeneous compounds, which supported the accuracy and robustness of our developed methodology. This assay exhibits universal potential applications for water quality monitoring, crop pollution detection, food and drug safety evaluation, as well as chemical warfare agent detection.

Abstract Mutations in the FERM domain containing 7 (FRMD7) gene have been proven to be responsible for infantile nystagmus (IN). The purpose of this study is to investigate FRMD7 gene mutations in patients with IN, and to evaluate the nystagmus intensity among patients with and without FRMD7 mutations. The affected males were subdivided into three groups according to whether or not having FRMD7 mutations and the types of mutations. Fifty-two mutations were detected in FRMD7 in 56 pedigrees and 34 sporadic patients with IN, including 28 novel and 24 previous reported mutations. The novel identified mutations further expand the spectrum of FRMD7 mutations. The parameters of nystagmus intensity and the patients’ best corrected visual acuity were not statistically different among the patients with and without identified FRMD7 mutations, and also not different among patients with different mutant types. The FERM-C domain, whose amino acids are encoded by exons 7, 8 and 9, could be the harbor region for most mutations. Loss-of-function is suggested to be the common molecular mechanism for the X-linked infantile nystagmus.

Purpose: This study aimed to explore the prevalence and predictors of incidental prostate cancer (IPC) after transurethral resection of the prostate (TURP) with negative results on transperineal magnetic resonance imaging (MRI)/transrectal ultrasonography (TRUS) fusion prostate biopsy or TRUS-guided prostate biopsy. Materials and Methods: Data of 253 patients who underwent TURP with a preliminary diagnosis of benign prostatic hyperplasia (BPH) were evaluated. The prevalence of IPC was calculated. Univariate and multivariate logistic regression analyses were conducted to explore independent predictive factors of IPC. Results: A total of 253 patients were included. IPC was diagnosed in 12 patients (4.7%). The mean age of the patients and the mean prostate volume were 69.8±7.07 years and 89.3±49.29 mL, respectively. The prevalence of IPC was higher in the TRUS guided prostate biopsy group than in the transperineal MRI/TRUS fusion prostate biopsy group (11 of 203 [5.4%] vs. 1 of 50 [2.0%], p=0.47), but the difference was not statistically significant. Our results indicated that older age (≥70 y) (odds ratio [OR], 1.14; 95% confidence interval [CI], 1.02–1.27; p=0.025) and smaller prostate volume (OR, 0.97; 95% CI, 0.938–0.998; p=0.039) were associated with an increased incidence of IPC after TURP. Conclusions: Our findings indicate that the prevalence of IPC may be higher among patients who undergo transrectal prostate biopsy before TURP than among those who undergo transperineal MRI/TRUS fusion prostate biopsy. Older age and smaller prostate volume were independent predictors of increasing the risk for IPC after TURP.

Abstract Background Polycyclic aromatic hydrocarbons (PAHs) pose a potential risk to ecological safety and human health. They have a range of effects on plant growth and there have been few reports on the health risks associated with ingestion of vegetable crops at different growth stages. Methodology In this study, a pot experiment in which Chinese cabbage (Brassica campestris L.) were grown in a greenhouse for 75 days was used to investigate the dose–effect relationship of pyrene with plant growth and also the exposure risk for adults of ingestion of Chinese cabbage at different growth stages. Results The results showed that low doses of pyrene (5–45 mg kg−1) promoted plant growth (20–220% and 55–97% higher than control treatment for the root biomass and shoot biomass, respectively), but significant inhibition was observed at a high dose (405 mg kg−1) (41–66% and 43–91% lower than control treatment for the root biomass and shoot biomass, respectively). High doses of pyrene reduced soil bacterial abundance and diversity during the growth of Chinese cabbage, and increased malondialdehyde (MDA) levels in the plant. The effects of pyrene on plant biomass were mainly attributed to changes in root activity induced by pyrene, as the relationship between soil pyrene concentration and biomass was similar to that between soil pyrene concentration and root activity. Furthermore, structural equation modeling analysis showed that pyrene altered growth of the vegetable by directly affecting root activity. The incremental lifetime cancer risk for adults is highest for ingestion of Chinese cabbage at the seedling stage, followed in decreasing order by the rosette stages and heading stages. Conclusions The health risk of consumers who have the possibility to ingest the Chinese cabbage planted in pyrene-contaminated soil would be decreased with the increasing growth periods. However, further studies are required to confirm the dose–effect relationship between pyrene concentration and Chinese cabbage growth on a field scale. Graphical Abstract

ObjectiveWe explored the DNA methylation and messenger RNA (mRNA) co-expression network and hub genes in first-episode, drug-naive adolescents with major depressive disorder (MDD). To preliminarily explore whether adolescent MDD has unique mechanisms compared with adult MDD.MethodsWe compared DNA methylation and mRNA profiles of peripheral blood mononuclear cells from four first-episode and drug-naive adolescents with MDD and five healthy adolescent controls (HCs). We performed differential expression analysis, constructed co-expression network, and screened the hub genes. And enrichment analysis was performed based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). We also downloaded DNA methylation and mRNA datasets of adults with MDD (GSE113725/GSE38206) from the GEO database, and performed differential expression and enrichment analysis.ResultsOur clinical data showed that 3034 methylation sites and 4190 mRNAs were differentially expressed in first-episode, drug-naive adolescents MDD patients compared with HCs. 19 hub genes were screened out according to the high degree value in the co-expression network. The results from the GEO database showed that compared with adult HCs, there were 290 methylation sites and 127 mRNAs were differentially expressed in adult MDD patients.ConclusionCompared with adolescent HCs and adult MDD patients, the DNA methylation and mRNA expression patterns of first-episode, drug-naive adolescent MDD patients were different. The co-expression network of DNA methylation and mRNA and the screened hub genes may play an important role in the pathogenesis of MDD in first-episode, drug-naive adolescents. Compared with adult MDD, adolescent MDD is more enriched in metabolism in terms of function and pathways.

The PAX6 gene plays an important role in ocular development. Mutations of the PAX6 gene may result in a series of ocular abnormalities, including congenital aniridia, anterior segment dysgenesis (ASD), progressive corneal opacification, glaucoma, and hypoplasia of the fovea and optic nerve, leading to reduced visual acuity and even blindness. This study aimed to describe the diversity of clinical features caused by PAX6 pathogenic variants in 45 Han Chinese patients from 23 unrelated families. All patients underwent detailed clinical assessment. Genetic testing was performed to identify pathogenic variations in the PAX6 gene by next-generation sequencing, minigene splicing assay, RT-qPCR, and long-range PCR. Twenty pathogenic variations were detected in the PAX6 gene from 12 pedigrees and 11 sporadic patients, of which 12 were previously reported and 8 were novel. The clinical phenotypes obtained as a result of the PAX6 gene mutations were complicated and vary among patients, even among those who carried the same variants. Genetic testing is helpful for differential diagnosis. Our genetic findings will expand the spectrum of pathogenic variations in the PAX6 gene. PAX6 pathogenic variants not only cause defects in ocular tissues, such as the iris and retina, but also lead to maldevelopment of the whole eye, resulting in microphthalmia.

Abnormal brain structural and functional properties were demonstrated in patients with infantile esotropia (IE). However, few studies have investigated the interaction between structural and functional connectivity (SC-FC) in patients with IE. Structural network was generated with diffusion tensor imaging and functional network was constructed with resting-state functional magnetic resonance imaging for 18 patients with IE as well as 20 age- and gender- matched healthy subjects. The SC-FC coupling for global connectome, short connectome and long connectome were examined in IE patients and compared with those of healthy subjects. A linear mixed effects model was employed to examine the group-age interaction in terms of the coupling metrics. The Pearson correlation between coupling measures and strabismus degree was evaluated in IE patients, on which the regulatory effect of age was also investigated through hierarchical regression analysis. Significantly decreased SC-FC coupling score for short connections was observed in left caudate nucleus (CAU) in IE patients, whereas no brain regions exhibited altered coupling metrics for global connections or long connections. The group-age interaction was also evident in local coupling metrics of left CAU. The age-related regulatory effect on coupling-degree association was distinguishing between brain regions implicated in visual processing and cognition-related brain areas in IE patients. Local SC-FC decoupling in CAU was evident in patients with IE and was initiated in their early postnatal period, possibly interfering the visual cortico-striatal loop and subcortical optokinetic pathway subserving visual processing and nasalward optokinesis during neurodevelopment, which provides new insight into underlying neuropathological mechanism of IE.

“Undruggable” targets such as KRAS are particularly challenging in the development of drugs. We devised a novel chemical knockdown strategy, CANDDY (Chemical knockdown with Affinity aNd Degradation DYnamics) technology, which promotes protein degradation using small molecules (CANDDY molecules) that are conjugated to a degradation tag (CANDDY tag) modified from proteasome inhibitors. We demonstrated that CANDDY tags allowed for direct proteasomal target degradation independent of ubiquitination. We synthesized a KRAS-degrading CANDDY molecule, TUS-007, which induced degradation in KRAS mutants (G12D and G12V) and wild-type KRAS. We confirmed the tumor suppression effect of TUS-007 in subcutaneous xenograft models of human colon cells (KRAS G12V) with intraperitoneal administrations and in orthotopic xenograft models of human pancreatic cells (KRAS G12D) with oral administrations. Thus, CANDDY technology has the potential to therapeutically target previously undruggable proteins, providing a simpler and more practical drug targeting approach and avoiding the difficulties in matchmaking between the E3 enzyme and the target.

Abstract Background The Coronavirus Disease 2019 (COVID-19) pandemic continues to threaten the physical and mental health of people across the world. This study aimed to understand the psychological impact of this disease on adolescents with major depressive disorder (MDD) at 1 month after the start of the outbreak in China. Methods Using the Children’s Impact of Event Scale (CRIES-13) questionnaire, we investigated the occurrence of posttraumatic stress disorder (PTSD) in two groups of adolescents: MDD patients who were in continuous antidepressant therapy and healthy controls. Total scores and factor subscores were compared between the two groups and subgroups stratified by sex and school grade. Logistic regression was used to identify variables associated with high total CRIES-13 scores. Results Compared to controls (n = 107), the MDD group (n = 90) had higher total CRIES-13 scores and a higher proportion with a total score ≥ 30. They also had a lower intrusion subscore and a higher arousal subscore. In the MDD group, males and females did not differ significantly in total CRIES-13 scores or factor subscores, but junior high school students had higher avoidance subscores than senior high school students. Logistic regression showed high total CRIES-13 scores to be associated with MDD and the experience of “flashbacks” or avoidance of traumatic memories associated with COVID-19. Conclusions It is crucial to understand the psychological impact of COVID-19 on adolescents with MDD in China, especially females and junior high school students. Long-term monitoring of adolescents with a history of mental illness is required to further understand these impacts. Trial registration ChiCTR, ChiCTR2000033402 , Registered 31 May 2020,

Abstract Background The outbreak of Coronavirus Disease 2019(COVID-19) caused psychological stress in Chinese adults population. But we are unaware of whether the pandemic causes psychological stress on children. Methods We used the Children’s Impact of Event Scale questionnaire (CRIES-13) to investigate the degree of Post-traumatic Stress (PTSD) symptoms caused by the pandemic in students selected from schools in Sichuan, Jiangsu, Henan, Yunnan, and Chongqing provinces of China. Results A total of 7769 students(3692 male and 4077 female), aged 8–18 years, were enrolled in the study, comprising 1214 in primary schools, 2799 in junior high schools and 3756 in senior high schools. A total of 1639 students (21.1%) had severe psychological stress reactions. A large proportion of senior high school students (23.3%) experienced severe psychological stress, and they had the highest median total CRIES-13 score. Female students were more likely to experience severe psychological stress and had higher median CRIES-13 total scores than males. Conclusion COVID-19 has placed psychological stresses on primary and secondary school students in China. These stresses are more likely to reach severe levels among female students and senior high school students.

BackgroundHorizontal gaze palsy with progressive scoliosis (HGPPS) is a rare disorder mainly involved in ocular movement and spinal development. It is caused by a roundabout guidance receptor 3 (ROBO3) gene mutation. This study aimed to describe the clinical features of six patients with HGPPS and investigate the corresponding ROBO3 gene mutations.MethodsPatients underwent detailed clinical and imaging examinations. Whole-exome sequencing was performed to detect nucleotide variations in the disease-causing genes of HGPPS.ResultsSix pathogenic variants were detected in the ROBO3 gene from six patients with HGPPS, including two novel compound heterozygous mutations, c.1447C > T (p.R483X) and c.2462G > C (p.R821P); c.1033G > C (p.V345L) and c.3287G > T (p.C1096F); a novel homozygous indel mutation, c.565dupC (p.R191Pfs*61); and a known missense mutation, c.416G > T (p.G139V). Patients with HGPPS had horizontal conjugated eye movement defects and scoliosis with variable degrees, as well as flattened pontine tegmentum and uncrossed corticospinal tracts on magnetic resonance imaging.ConclusionOur genetic findings will expand the spectrum of ROBO3 mutations and help inform future research on the molecular mechanism of HGPPS.

Background: Waardenburg syndrome (WS) is a rare genetic disorder characterized by congenital sensorineural hearing loss and pigmentary abnormalities of the hair, skin and eyes. However, exotropia is rarely reported. The purpose of this study is to describe the clinical characteristics of three sporadic patients with WS and congenital exotropia and to investigate the disease-causing genes for them.Methods: Patients underwent detailed physical and ocular examinations. Ocular alignment and binocular status were evaluated. DNA was extracted and whole exome sequencing was performed to detect the pathogenic variations in the disease-causing genes for WS. Cloning sequencing was carried out for those indel variations.Results: Three unrelated patients were diagnosed with Waardenburg syndrome and congenital exotropia. Four novel variants, including c.136delA (p.I46Sfs*64) and c.668G>T (p.R223L) in PAX3, c.709dupC (p.Q237Pfs*119) in COL11A2, c.426G>A (p.W142X) in SOX10 gene, were detected in this study.Conclusion: Simultaneous presence of congenital exotropia and WS in our patients is suggested that WS could be involved in malfunction in the multiple nerve systems. Our genetic study will expand the mutation spectrum of PAX3, COL11A2 and SOX10 genes, and is helpful for further study on the molecular pathogenesis of WS.

Previous studies have shown that functional networks are present at birth and change dynamically throughout infancy and early childhood. However, the status of functional connectivity is still poorly understood in patients with infantile esotropia (IE). The aim of this study is to investigate the developmental trends of functional connectivity in patients with IE during a critical period of growth and development. A total of 17 patients with IE (9 males and 8 females; mean age: 3.36 ± 2.03 years, age range: 0.67–6.36 years) and 20 healthy subjects matched for age and gender were recruited and underwent resting-state functional magnetic resonance imaging. The whole-brain functional network connectivity was analyzed for the IE group and healthy control group. A general linear model was applied to assess the group-age interaction in terms of the functional connectivity. The discrepancy between the two groups in functional connectivity trajectories was also quantified across age and exhibited by the quadratic parabolic model. There were significant group-age interactions between the visual network and the default mode network, the visual network and the sensorimotor network, the limbic network and the default mode network, and within the limbic network in the functional connectivity. A U-shaped tendency across age, with an “inflection point” ranging from 3.1 to 4.0 years of age was exhibited in the difference between functional connectivity trajectories of the IE patients and normal controls. Abnormality in functional network connectivity could present in IE patients at birth, exhibiting aberrant developmental patterns over time. An abnormal functional network could reduce the ability of the cortex in visual information processing, further reactivating the subcortical visual information processing system, which is probably the pathogenesis of IE. Three to four years after birth is the critical time window for children with IE to establish normal network connections in the brain. Early surgery during this period may be helpful for affected children to have an opportunity to approach the normal development trajectory as early as possible.

BackgroundThe ketogenic diet (KD) is increasingly used to treat drug-resistant epilepsy because of its favorable effect on seizure reduction. Patients with mitochondrial diseases tend to experience seizures. Therefore, this study aimed to test the efficacy of the KD on participants with mitochondrial diseases in a controlled trial.MethodsParticipants from fourteen clinical centers who were diagnosed with mitochondrial disease were semi-randomized to either the intervention (KD) or control group. The KD group followed a 3-month KD intervention, while the control group received a 1-month normal diet initially and then a 3-month KD intervention. The primary outcome measure was seizure reduction. Biomarker changes, cognitive impairments, and side effects were also recorded, if available.ResultA total of 33 participants were assigned to the KD (n = 22) and control groups (n = 11). In the KD group, 31.8% (7/22) of participants achieved ≥50% seizure reduction after 1 month of diet intervention, which increased to 40.9% (9/22) at 3 months. In the control group, only 18.2% (2/11) of the participants had ≥50% seizure reduction during the normal diet period. After the control group was transferred to the KD, 63.6% (7/11) of participants had >50% seizure reduction, and this rate increased to 72.7% (8/11) at 3 months. The KD also showed high efficacy in participants with mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) or pathogenic variants in mitochondrial DNA (mtDNA) (90% and 93.3% response rates, respectively). The most frequent side effects reported at the 3-month review were vomiting, cold, hyperlipidemia, and bloating.ConclusionThe KD is a safe and effective therapy for seizure control in mitochondrial diseases, especially MELAS and pathogenic variants of mtDNA. KD intervention can be considered in the management of these patients.

BackgroundAdolescents with immature mind and unstable emotional control are high-risk groups of non-suicidal self-injury (NSSI) behavior. We meta-analyzed the global prevalence of NSSI and prevalence of NSSI characteristics in a non-clinical sample of adolescents between 2010 and 2021.MethodsA systematic search for relevant articles published from January 1, 2010 to June 30, 2021 was performed within the scholarly database search engines of CBM, CNKI, VIP, Wanfang, PubMed, Web of Science, PsycINFO, and Embase. Eligibility criteria were as follows: provided cross-sectional data on the prevalence of NSSI; the subjects were non-clinical sample adolescents; and a clear definition of NSSI was reported. We used the following definiton of NSSI as our standard: the deliberate, self-inflicted destruction of body tissue, such as cutting, burning, and biting, without attempted suicide. The quality evaluation tool for cross-sectional studies recommended by the JBI was used. The global prevalence of NSSI was calculated based on the random-effects model by Comprehensive Meta-analysis version 3.0. Subgroup analyses were performed to compare the prevalence according to sex, living place, smoking or drinking history, and family structure.ResultsSixty-two studies involving 264,638 adolescents were included. The aggregate prevalence of NSSI among a non-clinical sample of adolescents was similar between over a lifetime (22.0%, 95% CI 17.9–26.6) and during a 12-month period (23.2%, 95% CI 20.2–26.5). Repetitive NSSI was more common than episodic NSSI (20.3% vs. 8.3%) but the frequency of mild injury (12.6%) was similar to that of moderate injury (11.6%). Multiple-method NSSI occurred slightly more often compared than one-method NSSI (16.0% vs. 11.1%). The top three types of NSSI in adolescents were banging/hitting (12.0%, 95% CI 8.9–15.9), pinching (10.0%, 95% CI 6.7–14.8), and pulling hair (9.8%, 95% CI 8.3–11.5), and the least common type was swallowing drugs/toxic substances/chemicals (1.0%, 95% CI 0.5–2.2). Subgroup analyses showed that being female, smoking, drinking, having siblings, and belonging to a single-parent family may be linked to higher prevalence of NSSI.ConclusionThis meta-analysis found a high prevalence of NSSI in non-clinical sample of adolescents, but there are some changes in severity, methods, and reasons. Based on the current evidence, adolescents in modern society are more inclined to implement NSSI behavior by a variety of ways, which usually are repetitive, and moderate and severe injuries are gradually increasing. It is also worth noting that adolescents with siblings or in single-parent families are relatively more likely to implement NSSI behavior due to maladjustment to the new family model. Future research needs to continue to elucidate the features and risk factors of NSSI so as to intervene in a targeted way.LimitationThe limitation of this study is that the heterogeneity among the included studies is not low, and it is mainly related to Chinese and English studies. The results of this study should be used with caution.Systematic review registration[www.crd.york.ac.uk/prospero/], identifier [CRD42022283217].

Abstract Background The incidence of ischemic stroke in the context of vascular disease is high, and the expression of growth-associated protein-43 (GAP43) increases when neurons are damaged or stimulated, especially in a rat model of middle cerebral artery occlusion/reperfusion (MCAO/R). Experimental design We bioengineered neuron-targeting exosomes (Exo) conjugated to a monoclonal antibody against GAP43 (mAb GAP43) to promote the targeted delivery of quercetin (Que) to ischemic neurons with high GAP43 expression and investigated the ability of Exo to treat cerebral ischemia by scavenging reactive oxygen species (ROS). Results Our results suggested that Que loaded mAb GAP43 conjugated exosomes (Que/mAb GAP43-Exo) can specifically target damaged neurons through the interaction between Exo-delivered mAb GAP43 and GAP43 expressed in damaged neurons and improve survival of neurons by inhibiting ROS production through the activation of the Nrf2/HO-1 pathway. The brain infarct volume is smaller, and neurological recovery is more markedly improved following Que/mAb GAP43-Exo treatment than following free Que or Que-carrying exosome (Que-Exo) treatment in a rat induced by MCAO/R. Conclusions Que/mAb GAP43-Exo may serve a promising dual targeting and therapeutic drug delivery system for alleviating cerebral ischemia/reperfusion injury.