The Expression of Circ-Astn1 Inhibits High Glucose Induced Endothelial Progenitor Cell Dysfunction by Activating Autophagy.

BACKGROUND: Diabetes mellitus (DM) and complications such as chronic kidney disease and cardiovascular symptoms pose a substantial public health burden. Increasing studies have shown that circular RNAs (circRNAs) regulate many gene expressions that are essential in diverse pathological and biological procedures. However, the roles of particular circRNAs in DM are unclear. METHODS: In the current investigation, endothelial progenitor cells (EPCs) were used to search for abnormal expression of circRNAs by using high-throughput sequencing under high glucose (HG) conditions. The regulatory mechanisms and targets were then studied through bioinformatics analysis, luciferase reporter analysis, angiogenic differentiation experiments, flow cytometry detection of apoptosis and RT-qPCR analysis. RESULTS: The circ-Astn1 expression in EPCs decreased after HG treatment. Overexpression or circ-Astn1 suppressed HG induced endothelial cell damage. MicroRNA (miR)-138-5p and SIRT5 were found to be the downstream targets of circ-Astn1 through luciferase reporter analysis. SIRT5 downregulation or miR-138-5p overexpression reversed circ-Astn1's protective effect against HG induced endothelial cell dysfunction, including apoptosis and abnormal vascular differentiation. Furthermore, circ-Astn1 overexpression promoted autophagy activation by increasing SIRT5 expression under HG conditions. Our findings suggest that circ-Astn1 mediated promotion of SIRT5 facilitates autophagy by sponging miR-138-5p. CONLUSION: Together, our findings show that the overexpression of circ-Astn1 suppresses HG induced endothelial cell damage by targeting miR-138-5p/SIRT5 axis.

The effect of planned active play on the fundamental movement skills of preschool children.

BACKGROUND: At present, Chinese children aged 3-6 years old are facing challenges such as insufficient physical activity, declining physical health, and obesity, and China has yet to issue curriculum standards or physical activity guidelines for this age group. At the same time, the present kindergarten physical activity curriculum is insufficient. To address this issue, this study focused on designing and executing a planned active play intervention program for the kindergarten setting to analyze its efficacy in enhancing children's fundamental movement skills (FMS). This study aims to provide a reference for the theoretical and practical exploration of children's acquisition of fundamental movement skills in the Chinese context. METHODS: Fifty-two preschoolers participated in this study and were either part of an intervention group (n = 30) or a control group (n = 24). Children's FMS were assessed before and after the intervention using the Test of Gross Motor Development-3 (TGMD-3) and the balance ability of the The Movement Assessment Battery for Children-2 (MABC-2). Physical activity (PA) during the planned active play intervention and the routine physical activity curriculum were assessed using the SOFIT throughout the intervention. RESULTS: All the children significantly improved their locomotor skills, ball skills, and TGMD from baseline to the late assessment (p < 0.05). Children in the planned active play intervention group demonstrated greater rates of change (p < 0.001) and scored higher on ball skills and TGMD in the late assessment than those in the control group (p < 0.001). The children in the intervention group, but not those in the control group, significantly improved their balance over time (p < 0.05), and the former had greater rates of change (p < 0.001). Similarly, planned active play was found to provide children with more physical activity than the routine physical activity curriculum. CONCLUSIONS: The eight-week planned active play intervention was effective in improving FMS in preschool children, with higher rates of FMS change in children who completed the intervention than children in the control group.

Construction of a redox-active metal-organic framework with an octanuclear lithium one-dimensional building block.

A stable lithium metal-organic framework, constructed using a redox-active N,N,N',N'-tetrakis(4-carboxyphenyl)-1,4-phenylenediamine linker and Li8 cluster-based one-dimensional rod secondary building unit, exhibits good stability and reversible redox activity. The Li8-MOF, which can be oxidized by AgNO3, has the potential to function as an electrochromic device, thereby advancing the development of smart MOF materials.

Kinship analysis and pedigree reconstruction by RAD sequencing in cattle.

Kinship and pedigree, used for estimating inbreeding, heritability, selection, and gene flow, are useful for breeding and animal conservation. However, as the size of crossbred populations increases, inaccurate generation and parentage assignment in livestock farms increase. Restriction-site-associated DNA sequencing is a cost-effective platform for single nucleotide polymorphism (SNP) discovery and genotyping. Here, we performed a kinship analysis and pedigree reconstruction for Angus and Xiangxi yellow cattle. A total of 975 cattle, including 923 offspring with 24 known sires and 28 known dams, were sampled and subjected to SNP discovery and genotyping. The identified SNP panel included 7,305 SNPs capturing the maximum difference between paternal and maternal genome information, allowing us to distinguish F1 from F2 generations with 90% accuracy. In conclusion, we provided a low-cost and efficient SNP panel for kinship analyses and the improvement of local genetic resources, which are valuable for breed improvement, local resource utilization, and conservation.

Altered functional connectivity in patients with post-stroke fatigue: A resting-state fMRI study.

BACKGROUND: Post-stroke fatigue (PSF) was a common complication after stroke. This study aimed to explore the neuroimaging mechanism of PSF, which was rarely studied. METHODS: Patients with the first episode of ischemic stroke were recruited from the First Affiliated Hospital of Wenzhou Medical University between March 2021 and December 2022. The fatigue severity scale (FSS) was used to assess fatigue symptoms. PSF was diagnosed by a neurologist based on the FSS score and PSF diagnostic criteria. All the patients were scanned by resting-state functional MRI (rs-fMRI). Precuneus, the posterior node of default-mode network (pDMN), was related to fatigue. Therefore, imaging data were further analyzed by the seed-based resting-state functional connectivity (FC) approach, with the left (PCUN.L) and right precuneus (PCUN.R) being the seeds. RESULTS: A total of 70 patients with acute ischemic stroke were finally recruited, comprising 40 patients with PSF and 30 patients without PSF. Both the PCUN.L and PCUN.R seeds (pDMN) exhibited decreased FC with the prefrontal lobes located at the anterior part of DMN (aDMN), and the FC values were negatively correlated with FSS scores (both p < 0.001). These two seeds also exhibited increased FC with the right insula, and the FC values were positively correlated with FSS scores (both p < 0.05). CONCLUSION: The abnormal FC between the aDMN and pDMN was associated with PSF. Besides, the insula, related to interoception, might also play an important role in PSF.

Non-invasive diagnosis of pulmonary nodules by circulating tumor DNA methylation: A prospective multicenter study.

BACKGROUND: With the popularization of computed tomography, more and more pulmonary nodules (PNs) are being detected. Risk stratification of PNs is essential for detecting early-stage lung cancer while minimizing the overdiagnosis of benign nodules. This study aimed to develop a circulating tumor DNA (ctDNA) methylation-based, non-invasive model for the risk stratification of PNs. METHODS: A blood-based assay ("LUNG-TRAC") was designed to include novel lung cancer ctDNA methylation markers identified from in-house reduced representative bisulfite sequencing data and known markers from the literature. A stratification model was trained based on 183 ctDNA samples derived from patients with benign or malignant PNs and validated in 62 patients. LUNG-TRAC was further single-blindly tested in a single- and multi-center cohort. RESULTS: The LUNG-TRAC model achieved an area under the curve (AUC) of 0.810 (sensitivity = 74.4 % and specificity = 73.7 %) in the validation set. Two test sets were used to evaluate the performance of LUNG-TRAC, with an AUC of 0.815 in the single-center test (N = 61; sensitivity = 67.5 % and specificity = 76.2 %) and 0.761 in the multi-center test (N = 95; sensitivity = 50.7 % and specificity = 80.8 %). The clinical utility of LUNG-TRAC was further assessed by comparing it to two established risk stratification models: the Mayo Clinic and Veteran Administration models. It outperformed both in the validation and the single-center test sets. CONCLUSION: The LUNG-TRAC model demonstrated accuracy and consistency in stratifying PNs for the risk of malignancy, suggesting its utility as a non-invasive diagnostic aid for early-stage peripheral lung cancer. CLINICAL TRIAL REGISTRATION: www. CLINICALTRIALS: gov (NCT03989219).

iRGD-Guided Silica/Gold Nanoparticles for Efficient Tumor-Targeting and Enhancing Antitumor Efficacy Against Breast Cancer.

Background: Breast cancer presents significant challenges due to the limited effectiveness of available treatments and the high likelihood of recurrence. iRGD possesses both RGD sequence and C-terminal sequence and has dual functions of targeting and membrane penetration. iRGD-modified nanocarriers can enhance drug targeting of tumor vascular endothelial cells and penetration of new microvessels, increasing drug concentration in tumor tissues. Methods: The amidation reaction was carried out between SiO2/AuNCs and iRGD/PTX, yielding a conjugated drug delivery system (SiO2/AuNCs-iRGD/PTX, SAIP@NPs). The assessment encompassed the characterization of the morphology, particle size distribution, physicochemical properties, in vitro release profile, cytotoxicity, and cellular uptake of SAIP@NPs. The tumor targeting and anti-tumor efficacy of SAIP@NPs were assessed using a small animal in vivo imaging system and a tumor-bearing nude mice model, respectively. The tumor targeting and anti-tumor efficacy of SAIP@NPs were assessed utilizing a small animal in vivo imaging system and an in situ nude mice breast cancer xenograft model, respectively. Results: The prepared SAIP@NPs exhibited decent stability and a certain slow-release effect in phosphate buffer (PBS, pH 7.4). In vitro studies had shown that, due to the dual functions of transmembrane and targeting of iRGD peptide, SAIP@NPs exhibited strong binding to integrin αvß3, which was highly expressed on the membrane of MDA-MB-231 cells, improving the uptake capacity of tumor cells, inhibiting the rapid growth of tumor cells, and promoting tumor cell apoptosis. The results of animal experiments further proved that SAIP@NPs had longer residence time in tumor sites, stronger anti-tumor effect, and no obvious toxicity to major organs of experimental animals. Conclusion: The engineered SAIP@NPs exhibited superior functionalities including efficient membrane permeability, precise tumor targeting, and imaging, thereby significantly augmenting the therapeutic efficacy against breast cancer with a favorable safety profile.

MeIS: DNA Methylation-Based Immune Response Signatures for Thyroid Nodule Diagnostics.

CONTEXT: Accurately distinguishing between benign thyroid nodules (BTNs) and papillary thyroid cancers (PTCs) with current conventional methods poses a significant challenge. OBJECTIVE: We identify DNA methylation markers of immune response-related genes for distinguishing BTNs and PTCs. METHODS: In this study, we analyzed a public reduced representative bisulfite sequencing dataset and revealed distinct methylation patterns associated with immune signals in PTCs and BTNs. Based on these findings, we developed a diagnostic classifier named the Methylation-based Immune Response Signature (MeIS), which was composed of 15 DNA methylation markers associated with immune response-related genes. We validated MeIS's performance in 2 independent cohorts: Z.S.'s retrospective cohort (50 PTC and 18 BTN surgery-leftover samples) and Z.S.'s preoperative cohort (31 PTC and 30 BTN fine-needle aspiration samples). RESULTS: The MeIS classifier demonstrated significant clinical promise, achieving areas under the curve of 0.96, 0.98, 0.89, and 0.90 in the training set, validation set, Z.S.'s retrospective cohort, and Z.S.'s preoperative cohort, respectively. For the cytologically indeterminate thyroid nodules, in Z.S.'s retrospective cohort, MeIS exhibited a sensitivity of 91% and a specificity of 82%; in Z.S.'s preoperative cohort, MeIS achieved a sensitivity of 84% and a specificity of 74%. Additionally, combining MeIS and BRAF V600E detection improved the detecting performance of cytologically indeterminate thyroid nodules, yielding sensitivities of 98% and 87%, and specificities of 82% and 74% in Z.S.'s retrospective cohort and Z.S.'s preoperative cohort, respectively. CONCLUSION: The 15 markers we identified can be employed to improve the diagnostic of cytologically indeterminate thyroid nodules.

Renal function affects the safety and efficacy of tirofiban in acute ischemic stroke thrombectomy patients.

BACKGROUND: Acute ischemic stroke poses a significant health threat, and thrombectomy has become a routine treatment. Tirofiban has emerged as a promising adjunct therapy to minimize reocclusion after thrombectomy. We aimed to investigate whether renal function influences the safety and efficacy of tirofiban in patients undergoing endovascular therapy. METHODS: Patients' clinical data collected from the stroke unit were analyzed. The modified Rankin scale score and symptomatic intracranial hemorrhage (sICH) were used as outcome measures. RESULTS: A total of 409 patients (mean age: 66.5 years, 292 males [71.4%]) were included. Tirofiban significantly improved 3-month functional outcomes (adjusted odds ratio [aOR] = 2.408, 95% confidence interval [CI] 1.120-5.175), reduced 3-month mortality (aOR = 0.364, 95% CI 0.155-0.856), and decreased the incidence of sICH (aOR = 0.339, 95% CI 0.149-0.767) in patients with estimated glomerular filtration rate (eGFR) ≥ 90 mL/min/1.73 m². However, no significant improvement in prognosis was observed with tirofiban in patients with eGFR < 90 mL/min/1.73 m². Interaction analysis suggested a potential influence of renal function on tirofiban efficacy. CONCLUSION: Renal function may impact the efficacy of tirofiban. Administration of tirofiban in direct thrombectomy patients with normal renal function is safe and improves prognosis. However, the prognostic benefits of tirofiban are limited in patients with impaired renal function.

Efficacy of scalp stimulation for multidomain cognitive impairment in patients with post-stroke cognitive impairment and dementia: A network meta-analysis and meta-regression of moderators.

BACKGROUND: Scalp stimulation has gained more traction for post-stroke cognitive impairment and dementia (PSCID); the interaction between stimulation targets and parameters influences the response to the stimulation. However, the most efficacious treatment for improving different domains of cognitive impairment remains unknown. OBJECTIVE: We aimed to conduct a systematic review and network meta-analysis (NMA) to compare the efficacy of various scalp stimulation protocols used in PSCID treatment. METHODS: Randomized controlled trials of scalp stimulation in patients with PSCID were searched in eight databases over the past 20 years. Standardized mean differences (SMDs) for global and subdomain cognitive scores were pooled in Bayesian NMA. Moderators were examined using meta-regression analysis. RESULTS: A total of 90 trials, with 6199 patients, were included. Low-frequency repetitive transcranial magnetic stimulation (rTMS) over the unaffected dorsolateral prefrontal cortex (DLPFC) was highly suggested for alleviating global severity (SMD = 1.11, 95% CI (0.64, 1.57)). High-frequency rTMS over the left DLPFC was recommended for language use (1.85 (1.18, 2.52)), executive function (0.85 (0.36, 1.33)), orientation deficits (0.59 (0.07, 1.13)), and attention (0.85 (0.27, 1.43)). Anodal transcranial direct current stimulation over the affected DLPFC (2.03 (0.72, 3.34)) was recommended for treating memory impairment. Meta-regression analyses showed significant associations within attention, language and orientation. CONCLUSION: Overall, different cognitive domains have different optimal scalp stimulation prescriptions, and activating the affected key brain regions and inhibiting the unaffected area is still the most effective treatment.

Optimized Exon-Exon Junction Library and its Application on Rodents' Brain Transcriptome Analysis

ABSTRACT Background: Alternative splicing (AS), which plays an important role in gene expression and functional regulation, has been analyzed on genome-scale by various bioinformatic approaches based on RNA-seq data. Compared with the huge number of studies on mouse, the AS researches approaching the rat, whose genome is intermedia between mouse and human, were still limited. To enrich the knowledge on AS events in rodents' brain, we perfomed a comprehensive analysis on four transcriptome libraries (mouse cerebrum, mouse cerebellum, rat cerebrum, and rat cerebellum), recruiting high-throughput sequencing technology. An optimized exon-exon junction library approach was introduced to adapt the longer RNA-seq reads and to improve mapping efficiency. Results: In total, 7,106 mouse genes and 2,734 rat genes were differentially expressed between cerebrum and cerebellum, while 7,125 mouse genes and 1,795 rat genes exhibited varieties on transcript variant level. Only half of the differentially expressed exon-exon junctions could be reflected at gene expression level. Functional cluster analysis showed that 32 pathways in mouse and 9 pathways in rat were significantly enriched, and 6 of them were in both. Interestingly, some differentially expressed transcript variants did not show difference on gene expression level, such as PLCβ1 and Kcnma1. Conclusion: Our work provided a case study of a novel exon-exon junction strategy to analyze the expression of genes and isoforms, helping us understand which transcript contributes to the overall expression and further functional change.