bluuebunny/crossref_metadata_2025_split huggingface dataset:
Establishment of a Dihydrofolate Reductase Gene Knock-In Zebrafish Strain to Aid Preliminary Analysis of Congenital Heart Disease Mechanisms
Authors: Gong Ke , Xie Ting , Yang Yifeng , Luo Yong , Deng Yun , Chen Kun , Tan Zhiping , Guo Hui , Xie Li 
Published: December 2021

Background: The dihydrofolate reductase (DHFR) gene is imperative in development, therefore it is essential to explore its effects on heart development. Thus, here a dhfr zebrafish knock-in (KI) strain was constructed.Methods: CRISPR/Cas9 technology was used to establish the dhfr KI zebrafish strain. This strain was hybridized with TgG fluorescent strain zebrafish to observe the phenotypes of heart shape, size, and circularization direction. Wild-type (WT) and KI zebrafish were then dissected and histologically stained to observe pathological changes. Western blot analysis was used to verify the increased expressions of zebrafish genes after KI. Hybridization experiments were used to confirm the presence of abnormal gonadal dysplasia.Results: The zebrafish dhfr KI strain was successfully constructed through CRISPR/Cas9 technology. At 6 days post fertilization (dpf), microscopic examinations of KI (homozygous) specimens revealed pericardial effusions, heart compressions, and curled tails. Compared with WT, the Hematoxylin and Eosin (H&amp;E) tissue sections of KI-homozygous zebrafish showed defects such as reduced atria and ventricles. Western blot analysis indicated that the expression of the DHFR protein increased in both heterozygotes and homozygotes of dhfr KI zebrafish. Hybridization experiments revealed that dhfr KI may affect gonadal function.Conclusion: The DHFR gene plays an important regulatory role in the process of heart development, and copy number variations (CNVs) of this gene may constitute a new pathogenic mechanism of congenital heart disease (CHD).
Liraglutide Inhibits Osteoclastogenesis and Improves Bone Loss by Downregulating Trem2 in Female Type 1 Diabetic Mice: Findings From Transcriptomics
Authors: Yu Jie , Shi Yan-Chuan , Ping Fan , Li Wei , Zhang Hua-Bing , He Shu-Li , Zhao Yuan , Xu Ling-Ling , Li Yu-Xiu 
Published: December 2021

BackgroundThe mechanisms of bone fragility in type 1 diabetes (T1D) are not fully understood. Whether glucagon-like peptide-1 receptor (GLP-1R) agonists could improve bone quality in T1D context also remains elusive.AimsWe aimed to explore the possible mechanisms of bone loss in T1D and clarify whether liraglutide has effects on bone quality of T1D mice using transcriptomics.MethodsFemale streptozotocin-induced diabetic C57BL/6J mice were randomly divided into four groups and received the following treatments daily for 8 weeks: saline as controls, insulin, liraglutide, and liraglutide combined with insulin. These groups were also compared with non-STZ-treated normal glucose tolerance (NGT) group. Trunk blood and bone tissues were collected for analysis. Three tibia from each of the NGT, saline-treated, and liraglutide-treated groups were randomly selected for transcriptomics.ResultsCompared with NGT mice, saline-treated T1D mice manifested markedly hyperglycemia and weight loss, and micro-CT revealed significantly lower bone mineral density (BMD) and deficient microarchitectures in tibias. Eight weeks of treatment with liraglutide alone or combined with insulin rescued the decreased BMD and partly corrected the compromised trabecular microarchitectures. Transcriptomics analysis showed there were 789 differentially expressed genes mainly mapped to osteoclastogenesis and inflammation pathways. The RT-qPCR verified that the gene expression of Trem2, Nfatc1, Trap, and Ctsk were significantly increased in the tibia of T1D compared with those in the NGT group. Liraglutide treatment alone or combined with insulin could effectively suppress osteoclastogenesis by downregulating the gene expression of Trem2, Nfatc1, Ctsk, and Trap.ConclusionsTaken together, increased osteoclastogenesis with upregulated expression of Trem2 played an important role in bone loss of T1D mice. Liraglutide provided protective effects on bone loss in T1D mice by suppressing osteoclastogenesis.
A Novel Competing Endogenous RNA Network Associated With the Pathogenesis of Graves’ Ophthalmopathy
Authors: Yue Zifan , Mou Pei , Chen Sainan , Tong Fei , Wei Ruili 
Published: December 2021

Background: Growing evidence has recently revealed the characteristics of long noncoding (lncRNA)/circular RNA (circRNA)-microRNA (miRNA)-mRNA networks in numerous human diseases. However, a scientific lncRNA/circRNA-miRNA-mRNA network related to Graves’ ophthalmopathy (GO) remains lacking.Materials and methods: The expression levels of RNAs in GO patients were measured through high-throughput sequencing technology, and the results were proven by quantitative real-time PCR (qPCR). We constructed a protein-protein interaction (PPI) network using the Search Tool for the Retrieval of Interacting Genes (STRING) database and identified hub genes by the Cytoscape plug-in CytoHubba. Then, the miRNAs related to differentially expressed lncRNAs/circRNAs and mRNAs were predicted through seed sequence matching analysis. Correlation coefficient analysis was performed on the interesting RNAs to construct a novel competing endogenous RNA (ceRNA) network.Results: In total, 361 mRNAs, 355 circRNAs, and 242 lncRNAs were differentially expressed in GO patients compared with control patients, 166 pairs were identified, and ceRNA networks were constructed. The qPCR results showed that 4 mRNAs (THBS2, CHRM3, CXCL1, FPR2) and 2 lncRNAs (LINC01820:13, ENST00000499452) were differentially expressed between the GO patients and control patients.Conclusion: An innovative lncRNA/circRNA-miRNA-mRNA ceRNA network between GO patients and control patients was constructed, and two important ceRNA pathways were identified, the LINC01820:13-hsa-miR-27b-3p-FPR2 ceRNA pathway and the ENST00000499452-hsa-miR-27a-3p-CXCL1 pathway, which probably affect the autoimmune response and inflammation in GO patients.
Case Report: Glucocorticoid Effect Observation in a Ureteral Urothelial Cancer Patient With ICI-Associated Myocarditis and Multiple Organ Injuries
Authors: Hu Xiajun , Wei Yumiao , Shuai Xinxin 
Published: December 2021

Immune checkpoint inhibitor (ICI)-associated immune-related adverse events (irAEs) are becoming important safety issues worthy of attention despite the exciting therapeutic prospects. The growing development of new ICIs also brings new cases of irAEs, raising more challenges to clinicians. Cardiac injury is rare but life-threatening among diverse organ injuries, and effective interventions are critical for patients. Here, we report a novel programmed cell death protein-1 (PD-1) inhibitor tislelizumab-associated severe myocarditis and myositis accompanied by liver and kidney damage in a ureteral urothelial cancer patient, who was firstly treated by cardiologists because of cardiac symptoms. Due to the lack of experience about ICI-associated irAEs, an initial low-dose (0.5 mg/kg/day) and short-term methylprednisolone therapy was used and found to be ineffective and risky to the patient; then, steroid therapy was modulated to a higher dose (1.5 mg/kg/day) with prolonged time course, and improvement of patient symptoms and laboratory markers were observed quickly and persistently. The patient did not show adverse events under this steroid dosage. This case reports a rare tislelizumab-related myocarditis and multiple organ injuries, which provides valuable experience to cardiologists like us. Early recognition of ICI-associated myocarditis and sufficient dosage and time course of glucocorticoid therapy are critical for severe cases. High-quality clinical evidence about the precise diagnosis and therapy in ICI-associated myocarditis and other organ injuries are necessary to guide our clinical works.
Computational Prediction of Biomarkers, Pathways, and New Target Drugs in the Pathogenesis of Immune-Based Diseases Regarding Kidney Transplantation Rejection
Authors: Alfaro Rafael , Martínez-Banaclocha Helios , Llorente Santiago , Jimenez-Coll Victor , Galián José Antonio , Botella Carmen , Moya-Quiles María Rosa , Parrado Antonio , Muro-Perez Manuel , Minguela Alfredo , Legaz Isabel , Muro Manuel 
Published: December 2021

BackgroundThe diagnosis of graft rejection in kidney transplantation (KT) patients is made by evaluating the histological characteristics of biopsy samples. The evolution of omics sciences and bioinformatics techniques has contributed to the advancement in searching and predicting biomarkers, pathways, and new target drugs that allow a more precise and less invasive diagnosis. The aim was to search for differentially expressed genes (DEGs) in patients with/without antibody-mediated rejection (AMR) and find essential cells involved in AMR, new target drugs, protein-protein interactions (PPI), and know their functional and biological analysis.Material and MethodsFour GEO databases of kidney biopsies of kidney transplantation with/without AMR were analyzed. The infiltrating leukocyte populations in the graft, new target drugs, protein-protein interactions (PPI), functional and biological analysis were studied by different bioinformatics tools.ResultsOur results show DEGs and the infiltrating leukocyte populations in the graft. There is an increase in the expression of genes related to different stages of the activation of the immune system, antigenic presentation such as antibody-mediated cytotoxicity, or leukocyte migration during AMR. The importance of the IRF/STAT1 pathways of response to IFN in controlling the expression of genes related to humoral rejection. The genes of this biological pathway were postulated as potential therapeutic targets and biomarkers of AMR. These biological processes correlated showed the infiltration of NK cells and monocytes towards the allograft. Besides the increase in dendritic cell maturation, it plays a central role in mediating the damage suffered by the graft during AMR. Computational approaches to the search for new therapeutic uses of approved target drugs also showed that imatinib might theoretically be helpful in KT for the prevention and/or treatment of AMR.ConclusionOur results suggest the importance of the IRF/STAT1 pathways in humoral kidney rejection. NK cells and monocytes in graft damage have an essential role during rejection, and imatinib improves KT outcomes. Our results will have to be validated for the potential use of overexpressed genes as rejection biomarkers that can be used as diagnostic and prognostic markers and as therapeutic targets to avoid graft rejection in patients undergoing kidney transplantation.
Associations Between Physical Activity and Hypertension in Chinese Children: A Cross-Sectional Study From Chongqing
Authors: Wang Qianqian , Qu Ping , Chen Jingyu , Tang Xian , Hao Guang , Liang Xiaohua 
Published: December 2021

Background: Childhood blood pressure is a marker for cardiovascular disease risk later in life. Few studies examined the association between physical activity (PA) and hypertension in Chinese children, and this study aimed to explore this relationship.Methods: A cross-sectional study among pupils was conducted in Chongqing in 2014. In total, 15,203 children aged 6–12 years in Chongqing were included in this study. The duration of self-reported PA on school days and the weekend in pupils were collected with a standardized questionnaire.Results: The results showed that, on school days, only 22.3% of boys and 17.8% of girls engaged in more than 60 min of PA per day; while on the weekend, only 38.5% of boys and 32.0% of girls engaged in at least 60 min of PA per day. There was no strong evidence for an association between PA and systolic or diastolic hypertension in boys. However, in girls, a weak negative association between PA on weekdays and hypertension was observed, and there was a significant interactive effect of PA and obesity on hypertension risk (Pfor interaction = 0.042). In obese children, PA was positively related to the risk of hypertension.Conclusion: The overall level of PA among pupils in Chongqing was insufficient, and a weak negative relationship between PA and hypertension was found in girls.
Cardiovascular Autonomic Function Changes and Predictors During a 2-Year Physical Activity Program in Rheumatoid Arthritis: A PARA 2010 Substudy
Authors: Hupin David , Sarajlic Philip , Venkateshvaran Ashwin , Fridén Cecilia , Nordgren Birgitta , Opava Christina H. , Lundberg Ingrid E. , Bäck Magnus 
Published: December 2021

Background: Chronic inflammation leads to autonomic dysfunction, which may contribute to the increased risk of cardiovascular diseases (CVD) in patients with rheumatoid arthritis (RA). Exercise is known to restore autonomic nervous system (ANS) activity and particularly its parasympathetic component. A practical clinical tool to assess autonomic function, and in particular parasympathetic tone, is heart rate recovery (HRR). The aim of this substudy from the prospective PARA 2010 study was to determine changes in HRR post-maximal exercise electrocardiogram (ECG) after a 2-year physical activity program and to determine the main predictive factors associated with effects on HRR in RA.Methods: Twenty-five participants performed physiotherapist-guided aerobic and muscle-strengthening exercises for 1 year and were instructed to continue the unsupervised physical activity program autonomously in the next year. All participants were examined at baseline and at years 1 and 2 with a maximal exercise ECG on a cycle ergometer. HRR was measured at 1, 2, 3, 4, and 5 min following peak heart rate during exercise. Machine-learning algorithms with the elastic net linear regression models were performed to predict changes in HRR1 and HRR2 at 1 year and 2 years of the PARA program.Results: Mean age was 60 years, range of 41–73 years (88% women). Both HRR1 and HRR2 increased significantly from baseline to year 1 with guided physical activity and decreased significantly from year 1 to year 2 with unsupervised physical activity. Blood pressure response to exercise, low BMI, and muscular strength were the best predictors of HRR1/HRR2 increase during the first year and HRR1/HRR2 decrease during the second year of the PARA program.Conclusion: ANS activity in RA assessed by HRR was improved by guided physical activity, and machine learning allowed to identify predictors of the HRR response at the different time points. HRR could be a relevant marker of the effectiveness of physical activity recommended in patients with RA at high risk of CVD. Very inactive and/or high CVD risk RA patients may get substantial benefits from a physical activity program.
Deep Learning With 18F-Fluorodeoxyglucose-PET Gives Valid Diagnoses for the Uncertain Cases in Memory Impairment of Alzheimer’s Disease
Authors: Zhang Wei , Zhang Tianhao , Pan Tingting , Zhao Shilun , Nie Binbin , Liu Hua , Shan Baoci ,   Alzheimer’s Disease Neuroimaging Initiative
Published: December 2021

Objectives:Neuropsychological tests are an important basis for the memory impairment diagnosis in Alzheimer’s disease (AD). However, multiple memory tests might be conflicting within-subjects and lead to uncertain diagnoses in some cases. This study proposed a framework to diagnose the uncertain cases of memory impairment.Methods:We collected 2,386 samples including AD, mild cognitive impairment (MCI), and cognitive normal (CN) using 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) and three different neuropsychological tests (Mini-Mental State Examination, Alzheimer’s Disease Assessment Scale-Cognitive Subscale, and Clinical Dementia Rating) from the Alzheimer’s Disease Neuroimaging Initiative (ADNI). A deep learning (DL) framework using FDG-PET was proposed to diagnose uncertain memory impairment cases that were conflicting between tests. Subsequent ANOVA, chi-squared, andt-test were used to explain the potential causes of uncertain cases.Results:For certain cases in the testing set, the proposed DL framework outperformed other methods with 95.65% accuracy. For the uncertain cases, its positive diagnoses had a significant (p&lt; 0.001) worse decline in memory function than negative diagnoses in a longitudinal study of 40 months on average. In the memory-impaired group, uncertain cases were mainly explained by an AD metabolism pattern but mild in extent (p&lt; 0.05). In the healthy group, uncertain cases were mainly explained by a non-energetic mental state (p&lt; 0.001) measured using a global deterioration scale (GDS), with a significant depression-related metabolism pattern detected (p&lt; 0.05).Conclusion:A DL framework for diagnosing uncertain cases of memory impairment is proposed. Proved by longitudinal tracing of its diagnoses, it showed clinical validity and had application potential. Its valid diagnoses also provided evidence and explanation of uncertain cases based on the neurodegeneration and depression mental state.
Epigenetic Age Acceleration and Hearing: Observations From the Baltimore Longitudinal Study of Aging
Authors: Kuo Pei-Lun , Moore Ann Zenobia , Lin Frank R. , Ferrucci Luigi 
Published: December 2021

Objectives: Age-related hearing loss (ARHL) is highly prevalent among older adults, but the potential mechanisms and predictive markers for ARHL are lacking. Epigenetic age acceleration has been shown to be predictive of many age-associated diseases and mortality. However, the association between epigenetic age acceleration and hearing remains unknown. Our study aims to investigate the relationship between epigenetic age acceleration and audiometric hearing in the Baltimore Longitudinal Study of Aging (BLSA).Methods: Participants with both DNA methylation and audiometric hearing measurements were included. The main independent variables are epigenetic age acceleration measures, including intrinsic epigenetic age acceleration—“IEAA,” Hannum age acceleration—“AgeAccelerationResidualHannum,” PhenoAge acceleration—“AgeAccelPheno,” GrimAge acceleration—“AgeAccelGrim,” and methylation-based pace of aging estimation—“DunedinPoAm.” The main dependent variable is speech-frequency pure tone average. Linear regression was used to assess the association between epigenetic age acceleration and hearing.Results: Among the 236 participants (52.5% female), after adjusting for age, sex, race, time difference between measurements, cardiovascular factors, and smoking history, the effect sizes were 0.11 995% CI: (–0.00, 0.23), p = 0.054] for Hannum’s clock, 0.08 [95% CI: (–0.03, 0.19), p = 0.143] for Horvath’s clock, 0.10 [95% CI: (–0.01, 0.21), p = 0.089] for PhenoAge, 0.20 [95% CI: (0.06, 0.33), p = 0.004] for GrimAge, and 0.21 [95% CI: (0.09, 0.33), p = 0.001] for DunedinPoAm.Discussion: The present study suggests that some epigenetic age acceleration measurements are associated with hearing. Future research is needed to study the potential subclinical cardiovascular causes of hearing and to investigate the longitudinal relationship between DNA methylation and hearing.
Aberrant Dynamics of Regional Coherence Measured by Resting-State fMRI in Children With Benign Epilepsy With Centrotemporal Spikes (BECTS)
Authors: Jiang Lin , Ma Xuejin , Liu Heng , Wang Ji , Zhang Jiaren , Zhang Guoming , Li Shiguang , Zhang Tijiang 
Published: December 2021

Objective:To explore the dynamic features of intrinsic brain activity measured by fMRI in children with benign epilepsy with centrotemporal spikes (BECTS) and examine whether these indexes were associated with behaviors.Methods:We recruited 26 children with BECTS (10.35 ± 2.91 years) and 26 sex-, and age-matched (11.35 ± 2.51 years) healthy controls (HC) and acquired resting-state functional magnetic resonance imaging (rs-fMRI) and behavioral data. Dynamic regional homogeneity (dReHo), including mean and coefficient of variation (CV) metrics derived from the rs-fMRI data, and were compared between the BECTS and the HC groups.Results:Significantly decreased mean dReHo in bilateral supramarginal gyrus, left middle temporal gyrus (MTG.L), left postcentral gyrus and superior occipital gyrus were found in children with BECTS. Meanwhile, increased CV of dReHo in MTG.L and right fusiform in children with BECTS was revealed compared with HC. Further analyses of functional connectivity revealed decreased global signal FC existed in similar regions, linked with linguistic, social cognition, and sensorimotor processes, in children with BECTS compared with HCs. Moreover, the association analyses showed that the CV of dReHo in MTG.L was positively associated with age and a negative correlation was found between mean dReHo of MTG.L and disease duration. Besides, the CV of dReHo in MTG.L was found positively associated with the intelligence quotient (IQ) language scores and full IQ scores in children with BECTS, and the CV of dReHo in the left inferior temporal gyrus and Rolandic operculum were positively correlated with IQ operation scores and full IQ scores.Conclusion:Aberrant dynamic regional coherence in sensorimotor, linguistic, and lateral temporal regions suggests dynamical interplay that underlying cognitive performance in children with BECTS, suggesting an intrinsic dynamic mechanism for BECTS.
Potential Locations for Non-Invasive Brain Stimulation in Treating Schizophrenia: A Resting-State Functional Connectivity Analysis
Authors: Ning Yanzhe , Zheng Sisi , Feng Sitong , Zhang Binlong , Jia Hongxiao 
Published: December 2021

Introduction: Non-invasive brain stimulation (NIBS) techniques have been widely used for the purpose of improving clinical symptoms of schizophrenia. However, the ambiguous stimulation targets may limit the efficacy of NIBS for schizophrenia. Exploring effective stimulation targets may improve the clinical efficacy of NIBS in schizophrenia.Methods: We first conducted a neurosynth-based meta-analysis of 715 functional magnetic resonance imaging studies to identify schizophrenia-related brain regions as regions of interest. Then, we performed the resting-state functional connectivity analysis in 32 patients with first-episode schizophrenia to find brain surface regions correlated with the regions of interest in three pipelines. Finally, the 10–20 system coordinates corresponding to the brain surface regions were considered as potential targets for NIBS.Results: We identified several potential targets of NIBS, including the bilateral dorsal lateral prefrontal cortex, supplementary motor area, bilateral inferior parietal lobule, temporal pole, medial prefrontal cortex, precuneus, superior and middle temporal gyrus, and superior and middle occipital gyrus. Notably, the 10-20 system location of the bilateral dorsal lateral prefrontal cortex was posterior to F3 (F4), not F3 (F4).Conclusion: Conclusively, our findings suggested that the stimulation locations corresponding to these potential targets might help clinicians optimize the application of NIBS therapy in individuals with schizophrenia.