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Yan, W.

Publications and source records attributed to Yan, W..

107 records · Page 6Linked to original sources

FUNCTIONAL NETWORK CONNECTIVITY (FNC)-BASED GENERATIVE ADVERSARIAL NETWORK (GAN) AND ITS APPLICATIONS IN MENTAL DISORDERS DIAGNOSIS

Functional network connectivity (FNC) obtained from resting-state functional magnetic resonance imaging (fMRI) data have been commonly used to study mental disorders in neuroimaging applications. Likewise, generative adversarial networks (GANs) have performed well in multiple classification benchmark tasks. However, the application of GANs to fMRI is relatively rare. In this work, we proposed an FNC-based GAN for classifying brain disorders from healthy controls (HCs), in which FNC matrices were calculated by correlation of time courses derived from non-artefactual fMRI independent components (ICs). The proposed GAN model consisted of one discriminator (real FNCs) and one generator (fake FNCs), each has four fully-connected layers, and feature matching was implemented between each other to improve classification performance. An average accuracy of 70.1% with 10-fold cross-validation was achieved for classifying 269 major depressive disorder (MDD) patients from 286 HCs, at least 5.9% higher compared to other 6 popular classification approaches (54.5-64.2%). In another application to discriminating between 558 schizophrenia patients and 542 HCs from 7 sites, the proposed GAN model achieved 80.7% accuracy in leave-one-site-out prediction, outperforming support vector machine (SVM) and deep neural net (DNN) by 3-6%. To the best of our knowledge, this is the first attempt to apply GAN model based on fMRI data for mental disorder classification. Such a framework promises wide utility and great potential in neuroimaging biomarker identification.

bioengineering↗

Abnormal nitration and S-sulfhydration of Sp1-CSE-H2S pathway contribute to the progress of HHcy

Sp1 (Specificity protein 1)-CSE (cystathionine-{gamma}-lyase)-H2S (hydrogen sulfide) pathway plays an important role in homocysteine-metabolism, whose disorder can result in hyperhomocysteinemia. The deficiency of plasma H2S in patients and animal models with hyperhomocysteinemia has been reported but it is unclear whether this deficiency plays a role in the progress of hyperhomocysteinemia. Furthermore, it remains unknown whether the post-translational modification of Sp1 or CSE mediated by hyperhomocysteinemia itself can in turn affect the development of hyperhomocysteinemia. By both in vivo and in vitro studies, we conducted immunoprecipitation and maleimide assays to detect the post-translational modification of Sp1-CSE-H2S pathway and revealed four major findings: (1) the accumulation of homocysteine augmented the nitration of CSE, thus blunted its bio-activity and caused H2S deficiency. (2) H2S deficiency lowered the S-sulfhydration of Sp1 and inhibited its transcriptional activity, resulted in lower expression of CSE. CSE deficiency decreased the H2S level further, which in turn lowered the S-sulfhydration level of CSE. (3) CSE was S-sulfhydrated at Cys84, Cys109, Cys172, Cys229, Cys252, Cys307 and Cys310 under physiological conditions, mutation of Cys84, Cys109, Cys229, Cys252 and Cys307 decreased its S-sulfhydration level and bio-activity. (4) H2S deficiency could trap hyperhomocysteinemia into a progressive vicious circle and trigger a rapid increase of homocysteine, while blocking nitration or restoring S-sulfhydration could break this circle. In conclusion, this study reveals a novel mechanism involved in the disorder of homocysteine-metabolism, which may provide a candidate therapeutic strategy for hyperhomocysteinemia.

pathology↗

TRIM21 and PHLDA3 Negatively Regulate the Cross-Talk between the PI3K/AKT Pathway and PPP Metabolism

PI3K/AKT signaling is known to regulate cancer metabolism but whether metabolic pathway feedbacks and regulates the PI3K/AKT pathway is unclear. Here, we demonstrate the important reciprocal cross-talks between the PI3K/AKT signal and PPP branching metabolic pathways. PI3K/AKT activation stabilizes G6PD, the rate-limiting enzyme of PPP, by inhibiting a newly identified E3 ligase TIRM21, and promotes PPP. PPP metabolites, in turn, reinforce AKT activation and further promote cancer metabolic reprogramming by blocking the expression of an AKT inhibitor PHLDA3. Knockout TRIM21 or PHLDA3 promotes the cross-talks and cell proliferation. Importantly, PTEN null human cancer cells and in vivo murine models are sensitive to anti-PPP treatments, suggesting the importance of PPP in maintaining AKT activation even in the presence of a constitutively activated PI3K pathway. Our study suggests that blockade of these reciprocal cross-talks may have a therapeutic benefit for cancers with PTEN loss or PI3K/AKT activation.

cancer biology↗

Efficient multiplex genome editing using CRISPR-Mb3Cas12a in mice

Despite many advantages over Cas9, Cas12a has not been widely used in genome editing in mammalian cells largely due to its strict requirement of the TTTV protospacer adjacent motif (PAM) sequence. Here, we report that Mb3Cas12a (Moraxella bovoculi AAX11_00205) could edit the genome in murine zygotes independent of TTTV PAM sequences and with minimal on-target mutations and close to 100% editing efficiency when crRNAs of 23nt spacers were used.\n\nSummary statementCRISPR-Mb3Cas12a can target a broader range of sequences in murine zygotes compared to AsCas12a and LbCas12a, and has lower on-target effects than Cas9 and high overall knock-in efficiency.

genomics↗

Single-cell profiles of retinal neurons differing in resilience to injury reveal neuroprotective genes

Neuronal types in the central nervous system differ dramatically in their resilience to injury or insults. Here we studied the selective resilience of mouse retinal ganglion cells (RGCs) following optic nerve crush (ONC), which severs their axons and leads to death of ~80% of RGCs within 2 weeks. To identify expression programs associated with differential resilience, we first used single-cell RNA-seq (scRNA-seq) to generate a comprehensive molecular atlas of 46 RGC types in adult retina. We then tracked their survival after ONC, characterized transcriptomic, physiological, and morphological changes that preceded degeneration, and identified genes selectively expressed by each type. Finally, using loss- and gain-of-function assays in vivo, we showed that manipulating some of these genes improved neuronal survival and axon regeneration following ONC. This study provides a systematic framework for parsing type-specific responses to injury, and demonstrates that differential gene expression can be used to reveal molecular targets for intervention.

neuroscience↗

Mitochondrial dynamics quantitatively revealed by STED nanoscopy with an enhanced squaraine variant probe

Mitochondria play a critical role in generating energy to support the entire lifecycle of biological cells, yet it is still unclear how their morphological structures evolve to regulate their functionality. Conventional fluorescence microscopy can only provide [~]300 nm resolution, which is insufficient to visualize mitochondrial cristae. Here, we developed an enhanced squaraine variant dye (MitoESq-635) to study the dynamic structures of mitochondrial cristae in live cells at superresolution. The low saturation intensity and high photostability make it ideal for long-term, high-resolution STED nanoscopy. We demonstrate the time-lapsed imaging of the mitochondrial inner membrane over 50 minutes in living HeLa cells at 35.2 nm resolution for the first time. The forms of the cristae during mitochondrial fusion and fission can be clearly resolved. Our study demonstrates the emerging capability of optical STED nanoscopy to investigate intracellular physiological processes at nanoscale resolution for long periods of time with minimal phototoxicity.

biophysics↗

Precise removal of Calm1 long 3′ UTR isoform by CRISPR-Cas9 genome editing impairs dorsal root ganglion development in mice.

Most mammalian genes are subject to Alternative cleavage and PolyAdenylation (APA), often resulting in alternative length 3' UTR isoforms. Thousands of extended or long 3' UTR variants are preferentially expressed in neuron-enriched tissues of metazoans. However, the in vivo functions of these long 3' UTR isoforms are largely unknown. Calmodulin 1 (Calm1) is a key integrator of calcium signaling that is required for correct neural development. Calm1 generates short (Calm1-S) and long 3' UTR (Calm1-L) mRNA isoforms via APA. We found Calm1-S to be broadly expressed across mouse tissues, whereas Calm1-L expression was largely restricted to neural tissues, including the dorsal root ganglion (DRG). Using CRISPR-Cas9 genome editing, a series of mouse deletion lines were generated that successfully eliminated expression of Calm1-L while maintaining expression of Calm1-S. One of these lines, Calm1{Delta}3' UTR, carried a 163 bp deletion surrounding the distal polyA site. Examination of Calm1{Delta}3' UTR embryos revealed disrupted development of the DRG. In Calm1{Delta}3' UTR DRG explant cultures undergoing axon outgrowth, we observed a dramatic increase in axon fasciculation. These results demonstrate a physiological role for Calm1-L in DRG development, and more generally, establish a genome-editing strategy to study in vivo functions of long 3' UTR isoforms. Author SummaryMore than half of all human genes generate alternative mRNA isoforms which differ in the length of their 3 Untranslated regions (3 UTRs). Through a process called Alternative Cleavage and Polyadenylation thousands of broadly expressed genes preferentially express long 3 UTR variants in brain tissues whereas their short 3 UTR counterparts are more broadly expressed. A challenge to study the functions of these transcripts has been to generate loss of function mutant animals that lack a long 3 UTR isoform but maintain expression of the corresponding short 3 UTR isoform. Here, we used the precise, rapid, and efficient approach of CRISPR genome-editing to generate long 3 UTR mutant mice. These mice, which do not express the long 3 UTR of the Calmodulin 1 (Calm1) gene, exhibit impairment in the development of sensory neurons, including increased fasciculation of axons and aberrant cell body migration. This finding is important because it provides conclusive genetic evidence for a neural function of a long 3 UTR isoform in an animal. The CRISPR genome-editing approach used here can be applied to the study of neuron-enriched long 3 UTR isoforms, which number in the thousands and have largely unexplored functions.

genetics↗

Effects of Lactobacillus plantarum 15-1 and fructooligosaccharide on the response of broilers to pathogenic Escherichia coli O78 challenge

One-day-old broilers were randomly allocated to 5 treatments: basal diet challenged by saline (negative control, n-control); basal diet and challenged by E.coil O78 (positive control, p-control); supplementation with L. plantarum 15-1 at 1x108 CFU/kg challenged with E.coil O78 (LP); supplementation with FOS at 5 g/kg challenged with E.coil O78 (FOS); supplementation with L. plantarum 15-1 and FOS challenged with E.coil O78 (LP+FOS). L. plantarum 15-1 or FOS had a lowered effect (P<0.05) on crypt depth on d 14 compared with two controls, and L. plantarum 15-1, FOS and L. plantarum 15-1+FOS also reduced relative to p-control on d 21. L. plantarum 15-1 reduced the level of diamine oxidase (DAO) at d 14 and 21 compared with p-control (P<0.05), the broilers with L. plantarum 15-1 and FOS increased the concentration of IgA and IgG relative to two control, and decreased diamine oxidase (DAO) compared with p-control (P 0.05). L. plantarum 15-1 increased the concentration of acetic acid and total short-chain fatty acid (SCFA) in comparison with p-control at d 14 (P<0.05), FOS improved the level of valeric acid and total SCFA relative to p-control at d 21 (P<0.001), the broilers fed L. plantarum 15-1 and FOS were increased the level of butyric acid at d 14 (P<0.05). FOS enhanced bursal index of broilers at d 21 (P<0.05). L. plantarum 15-1 and FOS did no effect on the growth performance. In conclusion, FOS can promote average daily gain, L. plantarum 15-1 and FOS can improve intestinal morphology, and increase the concentration of SCFA in cecal contents in broilers challenged with E.coil O78. These results suggest that L. plantarum 15-1 and FOS have effective mitigation to E. coil O78 via lowing reducing the intestinal injury and enhancing the immune responses.

zoology↗

Multi-ATOM: Ultrahigh-throughput single-cell quantitative phase imaging with subcellular resolution

A growing body of evidence has substantiated the significance of quantitative phase imaging (QPI) in enabling cost-effective and label-free cellular assay, which provides useful insights into understanding biophysical properties of cells and their roles in cellular functions. However, available QPI modalities are limited by the loss of imaging resolution at high throughput and thus run short of sufficient statistical power at the single cell precision to define cell identities in a large and heterogeneous population of cells - hindering their utility in mainstream biomedicine and biology. Here we present a new QPI modality, coined multi-ATOM that captures and processes quantitative label-free single-cell images at ultra-high throughput without compromising sub-cellular resolution. We show that multi-ATOM, based upon ultrafast phase-gradient encoding, outperforms state-of-the-art QPI in permitting robust phase retrieval at a QPI throughput of >10,000 cell/sec, bypassing the need for interferometry which inevitably compromises QPI quality under ultrafast operation. We employ multi-ATOM for large-scale, label-free, multi-variate, cell-type classification (e.g. breast cancer sub-types, and leukemic cells versus peripheral blood mononuclear cells) at high accuracy (>94%). Our results suggest that multi-ATOM could empower new strategies in large-scale biophysical single-cell analysis with applications in biology and enriching disease diagnostics.

bioengineering↗

Molecular Classification and Comparative Taxonomics of Foveal and Peripheral Cells in Primate Retina

High acuity vision in primates, including humans, is mediated by a small central retinal region called the fovea. As more accessible model organisms lack a fovea, its specialized function and dysfunction in ocular diseases remain poorly understood. We used 165,000 single-cell RNA-seq profiles to generate and validate comprehensive cellular taxonomies of macaque fovea and peripheral retina. More than 80% of >65 cell types match between the two regions, but exhibit substantial differences in proportions and gene expression, some of which we relate to functional differences. Comparison of macaque retinal types with those of mice reveals that interneuron types are tightly conserved, but that projection neuron types and programs diverge, despite conserved transcription factor codes. Key macaque types are conserved in humans, allowing mapping of cell-type and region-specific expression of >190 genes associated with 6 human retinal diseases. Our work provides a framework for comparative single-cell analysis across tissue regions and species.

neuroscience↗

Active information maintenance in working memory by a sensory cortex

Working memory is a critical function of the brain to maintain and manipulate information over delay periods of seconds. Sensory areas have been implicated in working memory; however, it is debated whether the delay-period activity of sensory regions is actively maintaining information or passively reflecting top-down inputs. We hereby examined the anterior piriform cortex, an olfactory cortex, in head-fixed mice performing a series of olfactory working memory tasks. Information maintenance is necessary in these tasks, especially in a dual-task paradigm in which mice are required to perform another distracting task while actively maintaining information during the delay period. Optogenetic suppression of the piriform cortex activity during the delay period impaired performance in all the tasks.Furthermore, electrophysiological recordings revealed that the delay-period activity of the anterior piriform cortex encoded odor information with or without the distracting task.Thus, this sensory cortex is critical for active information maintenance in working memory.

neuroscience↗

Epidemiology, Microbiology and Therapeutic Consequences of Chronic Osteomyelitis in Northern China: A Retrospective Analysis of 255 Patients

The study aimed to explore the epidemiology and clinical characteristics of chronic osteomyelitis observed in a northern China hospital. Clinical data of 255 patients with chronic osteomyelitis from January 2007 to January 2014 were collected and analyzed, including general information, disease data, treatment and follow-up data. Chronic osteomyelitis is more common in males and in the age group from 41-50 years of age. Common infection sites are the femur, tibiofibular, and hip joint. More g+ than g- bacterial infections were observed, with S. aureus the most commonly observed pathogenic organism. The positive detection rate from debridement bacterial culture is 75.6%. The detection rate when five samples are sent for bacterial culture is 90.6%, with pathogenic bacteria identified in 82.8% of cases. The two-stage debridement method (87.0%) has higher first curative rate than the one-stage debridement method (71.2%). To improve detection rate using bacterial culture, at least five samples are recommended. Treatment of chronic osteomyelitis with two-stage debridement, plus antibiotic-loaded polymethylmethacrylate (PMMA) beads provided good clinical results in this study and is therefore recommended.

microbiology↗

Efficacy of fecal sampling as a gut proxy in the study of chicken gut microbiota

BackgroundDespite the convenience and noninvasiveness of fecal sampling, the fecal microbiota does not fully represent that of the gastrointestinal (GI) tract, and the efficacy of fecal sampling to accurately represent the gut microbiota in birds is poorly understood. In this study, we aim to identify the efficacy of feces as a gut proxy in birds using chickens as a model. We collected 1,026 samples from 206 chickens, including duodenum, jejunum, ileum, cecum and feces samples, for 16S rRNA amplicon sequencing analyses.\n\nResultsIn this study, the efficacy of feces as a gut proxy was partitioned to microbial community membership and community structure. Most taxa in the small intestine (84.11 - 87.28%) and ceca (99.39%) could be identified in feces. Microbial community membership was reflected with a gut anatomic feature, but community structure was not. Excluding shared microbes, the small intestine and ceca contributed 34.12 and 5.83% of the total fecal members, respectively. The composition of Firmicutes members in the small intestine and that of Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria members in the ceca could be well mirrored by the observations in fecal samples ({rho} = 0.54 - 0.71 and 0.71 - 0.78, respectively, P < 0.001). However, there were few significant correlations for each genus between feces and each of the 4 gut segments, and these correlations were not high ({rho} = -0.2 - 0.4, P < 0.05) for most genera.\n\nConclusionsOur results provide evidence that the good potential of feces to identify most taxa in chicken guts, but it should be interpreted with caution by using feces as a proxy for gut in microbial structure analyses. This work provides insights and future directions regarding the usage of fecal samples in studies of the gut microbiome.

microbiology↗

Template switching causes artificial junction formation and false identification of circular RNAs

Hundreds of thousands of putative circular RNAs have been identified through deep sequencing and bioinformatic analyses. However, the circularity of these putative RNA circles has not been experimentally validated due to limited methodologies currently available. We reported here that the template-switching capability of commonly used reverse transcriptases (e.g., SuperScript II) leads to the formation of artificial junction sequences, and consequently misclassification of large linear RNAs as RNA circles. Use of reverse transcriptases without terminal transferase activity (e.g., MonsterScript) for cDNA synthesis is critical for the identification of physiological circular RNAs. We also report two methods, MonsterScript junction PCR and high-resolution melting curve analyses, which can reliably distinguish circular RNAs from their linear forms and thus, can be used to discover and validate true circular RNAs.\n\nSignificance StatementThe vast majority of circular RNAs were identified through computational detection of junction sequences in the deep sequencing reads because these unique fusion sequences represent back-splicing events. We found that artificial junction sequences could be formed through template switching (TS) when MMLV-derived reverse transcriptases, e.g., SuperScript II, are used to synthesize cDNAs. Thus, many of the reported circular RNAs may not be RNA circles, but rather experimental artifacts. Fake circular RNAs can be avoided by using reverse transcriptases without terminal transferase activity (e.g., MonsterScript) for cDNA synthesis. We developed two novel methods, MonsterScript junction PCR and high-resolution melting curve analyses, for distinguishing circular RNAs from their linear form.

molecular biology↗

Whole genome sequence of an edible and potential medicinal fungus, Cordyceps guangdongensis

Cordyceps guangdongensis is an edible fungus which has been approved as a Novel Food by the Chinese Ministry of Public Health in 2013. It also has a broad application prospect in pharmaceutical industries with many medicinal activities. In this study, the whole genome of C. guangdongensis GD15, a single spore isolate from a wild strain, was sequenced and assembled with Illumina and PacBio sequencing technology. The generated genome is 29.05 Mb in size, comprising 9 scaffolds with an average GC content of 57.01%. It is predicted to contain a total of 9150 protein-coding genes. Sequence identification and comparative analysis indicated that the assembled scaffolds contained two complete chromosomes and four single-end chromosomes, showing a high level assembly. Gene annotation revealed a diversity of transporters that could contribute to the genome size and evolution. Besides, approximately 15.49% and 13.70% genes involved in metabolic processes were annotated by KEGG and COG respectively. Genes belonging to CAZymes accounted for a proportion of 2.84% of the total genes. In addition, 435 transcription factors (TFs) were identified, which were involved in various biological processes. Among the identified TFs, the fungal transcription regulatory proteins (18.39%) and fungal-specific TFs (19.77%) represented the two largest classes of TFs. These data provided a much needed genomic resource for studying C. guangdongensis, laying a solid foundation for further genetic and biological studies, especially for elucidating the genome evolution and exploring the regulatory mechanism of fruiting body development.

genomics↗

UV-Induced Keratin 1 Proteolysis Mediates UV-Induced Skin Damage

Keratins play critical roles in intermediate filament formation, inflammatory responses and cellular signaling in epithelium. While keratins is a major epidermal fluorophore, the mechanisms underlying the autofluorescence (AF) of keratins and its biomedical implications have remained unknown. Our study used mouse skin as a model to study these topics, showing that UV dose-dependently induced increases in green AF at the spinous layer of the epidermis of mouse within 6 hr of the UV exposures, which may be used for non-invasive prediction of UV-induced skin damage. The UV-induced AF appears to be induced by cysteine protease-mediated keratin 1 proteolysis: 1) UV rapidly induced significant keratin 1 degradation; 2) administration of keratin 1 siRNA largely decreased the UV-induced AF; and 3) administration of E-64, a cysteine protease inhibitor, significantly attenuated the UV-induced AF and keratin 1 degradation. Our study has also suggested that the UV-induced keratin 1 proteolysis may be a novel crucial pathological factor in UV-induced skin damage, which is supported by both the findings that indicate critical biological roles of keratin 1 in epithelium and our observation that prevention of UV-induced keratin 1 proteolysis can lead to decreased UV-induced skin damage. Collectively, our study has suggested that UV-induced keratin 1 proteolysis may be a novel and valuable target for diagnosis, prevention and treatment of UV-induced skin damage.

pathology↗

AASRA: An Anchor Alignment-Based Small RNA Annotation Pipeline

SncRNA-Seq has become a routine for sncRNA profiling; however, software packages currently available are either exclusively for miRNA or piRNA annotation (e.g., miRDeep, miRanalyzer, Shortstack, PIANO), or for direct mapping of the sequence reads to the genome (e.g., Bowtie 2, SOAP and BWA), which tend to generate inaccurate counting due to repetitive matches to the genome or sncRNA homologs. Moreover, novel sncRNA variants in the sequencing reads, including those bearing small overhangs or internal insertions, deletions or mutations, are totally excluded from counting by these algorithms, leading to potential quantification bias. To overcome these problems, a comprehensive software package that can annotate all known small RNA species with adjustable tolerance towards small mismatches is needed. AASRA is based on our unique anchor alignment algorithm, which not only avoids repetitive or ambiguous counting, but also distinguishes mature miRNA from precursor miRNA reads. Compared to all existing pipelines for small RNA annotation, AASRA is superior in the following aspects: 1) AASRA can annotate all known sncRNA species simultaneously with the capability of distinguishing mature and precursor miRNAs; 2) AASRA can identify and allow for inclusion of sncRNA variants with small overhangs and/or internal insertions/deletions into the final counts; 3) AASRA is the fastest among all small RNA annotation pipelines tested. AASRA represents an all-in-one sncRNA annotation pipeline, which allows for high-speed, simultaneous annotation of all known sncRNA species with the capability to distinguish mature from precursor miRNAs, and to identify novel sncRNA variants in the sncRNA-Seq sequencing reads.\n\nAvailability and ImplementationThe AASRA software is freely available at https://github.com/biogramming/AASRA.

bioinformatics↗