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1.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 55(2): 309-314, 2024 Mar 20.
Article in Chinese | MEDLINE | ID: mdl-38645869

ABSTRACT

Objective: To explore the application effect of intelligent health education based on the health belief model on patients with postoperative kinesophobia after surgical treatment of cervical spondylosis. Methods: A prospective cohort study was conducted with patients who underwent anterior cervical discectomy, decompression, and fusion surgery with a single central nerve and spine center, and who had postoperative kinesophobia, ie, fear of movement. The patients made voluntary decisions concerning whether they would receive the intervention of intelligent health education. The patients were divided into a control group and an intelligent education group and the intervention started on the second day after the surgery. The intelligent education group received intelligent education starting from the second day after surgery through a WeChat widget that used the health belief model as the theoretical framework. The intelligent health education program was designed according to the concept of patient problems, needs, guidance, practice, and feedbacks. It incorporated four modules, including knowledge, intelligent exercise, overcoming obstacles, and sharing and interaction. It had such functions as reminders, fun exercise, shadowing exercise, monitoring, and documentation. Health education for the control group also started on the second day after surgery and was conducted by a method of brochures of pictures and text and WeChat group reminder messages. The participants were surveyed before discharge and 3 months after their surgery. The primary outcome measure compared between the two groups was the degree of kinesophobia. Secondary outcome measures included differences in adherence to functional exercise (Functional Exercise Adherence Scale), pain level (Visual Analogue Scale score), degree of cervical functional impairment (Cervical Disability Index), and quality of life (primarily assessed by the Quality of Life Short Form 12 [SF-12] scale for psychological and physiological health scores). Results: A total of 112 patients were enrolled and 108 patients completed follow-up. Eventually, there were 53 cases in the intelligent education group and 55 cases in the control group. None of the patients experienced any sports-related injuries. There was no statistically significant difference in the primary and secondary outcome measures between the two groups at the time of discharge. At the 3-month follow-up after the surgery, the level of kinesophobia in the intelligent education group (25.72±3.90) was lower than that in the control group (29.67±6.16), and the difference between the two groups was statistically significant (P<0.05). In the intelligent education group, the degree of pain (expressed in the median [25th percentile, 75th percentile]) was lower than that of the control group (0 [0, 0] vs. 1 [1, 2], P<0.05), the functional exercise adherence was better than that of the control group (63.87±7.26 vs. 57.73±8.07, P<0.05), the psychological health was better than that of the control group (40.78±3.98 vs. 47.78±1.84, P<0.05), and the physical health was better than that of the control group (43.16±4.41 vs. 46.30±3.80, P<0.05), with all the differences being statistically significant. There was no statistically significant difference in the degree of cervical functional impairment between the two groups (1 [1, 2] vs. 3 [2, 7], P>0.05). Conclusion: Intelligent health education based on the health belief model can help reduce the degree of kinesophobia in patients with postoperative kinesophobia after surgical treatment of cervical spondylosis and improve patient prognosis.


Subject(s)
Cervical Vertebrae , Spondylosis , Humans , Spondylosis/surgery , Prospective Studies , Cervical Vertebrae/surgery , Phobic Disorders/psychology , Female , Male , Diskectomy/methods , Patient Education as Topic/methods , Decompression, Surgical/methods , Fear , Middle Aged , Health Education/methods , Spinal Fusion/methods , Kinesiophobia
2.
Appl Opt ; 63(6): A7-A15, 2024 Feb 20.
Article in English | MEDLINE | ID: mdl-38437352

ABSTRACT

Accurate and efficient counting of shrimp larvae is crucial for monitoring reproduction patterns, assessing growth rates, and evaluating the performance of aquaculture. Traditional methods via density estimation are ineffective in the case of high density. In addition, the image contains bright spots utilizing the point light source or the line light source. Therefore, in this paper an automated shrimp counting platform based on optics and image processing is designed to complete the task of counting shrimp larvae. First, an area light source ensures a uniformly illuminated environment, which helps to obtain shrimp images with high resolution. Then, a counting algorithm based on improved k-means and a side window filter (SWF) is designed to achieve an accurate number of shrimp in the lamp house. Specifically, the SWF technique is introduced to preserve the body contour of shrimp larvae, and eliminate noise, such as water impurities and eyes of shrimp larvae. Finally, shrimp larvae are divided into two groups, independent and interdependent, and counted separately. Experimental results show that the designed optical counting system is excellent in terms of visual effect and objective evaluation.


Subject(s)
Algorithms , Aquaculture , Animals , Eye , Image Processing, Computer-Assisted , Larva
3.
Nat Commun ; 15(1): 1025, 2024 Feb 03.
Article in English | MEDLINE | ID: mdl-38310199

ABSTRACT

RNA modification C2-methyladenosine (m2A) exists in both rRNA and tRNA of Escherichia coli (E. coli), installed by the methyltransferase RlmN using a radical-S-adenosylmethionine (SAM) mechanism. However, the precise function of m2A in tRNA and its ubiquity in plants have remained unclear. Here we discover the presence of m2A in chloroplast rRNA and tRNA, as well as cytosolic tRNA, in multiple plant species. We identify six m2A-modified chloroplast tRNAs and two m2A-modified cytosolic tRNAs across different plants. Furthermore, we characterize three Arabidopsis m2A methyltransferases-RLMNL1, RLMNL2, and RLMNL3-which methylate chloroplast rRNA, chloroplast tRNA, and cytosolic tRNA, respectively. Our findings demonstrate that m2A37 promotes a relaxed conformation of tRNA, enhancing translation efficiency in chloroplast and cytosol by facilitating decoding of tandem m2A-tRNA-dependent codons. This study provides insights into the molecular function and biological significance of m2A, uncovering a layer of translation regulation in plants.


Subject(s)
Arabidopsis , Escherichia coli , Escherichia coli/genetics , Escherichia coli/metabolism , RNA, Transfer/genetics , RNA, Transfer/metabolism , Methyltransferases/metabolism , Codon/genetics , S-Adenosylmethionine/metabolism , Plants/metabolism , RNA, Ribosomal/metabolism , Arabidopsis/genetics , Arabidopsis/metabolism , Protein Biosynthesis
4.
Angew Chem Int Ed Engl ; 63(7): e202313900, 2024 Feb 12.
Article in English | MEDLINE | ID: mdl-38158383

ABSTRACT

N1 -methyladenosine (m1 A) is a prevalent post-transcriptional RNA modification, and the distribution and dynamics of the modification play key epitranscriptomic roles in cell development. At present, the human AlkB Fe(II)/α-ketoglutarate-dependent dioxygenase family member ALKBH3 is the only known mRNA m1 A demethylase, but its catalytic mechanism remains unclear. Here, we present the structures of ALKBH3-oligo crosslinked complexes obtained with the assistance of a synthetic antibody crystallization chaperone. Structural and biochemical results showed that ALKBH3 utilized two ß-hairpins (ß4-loop-ß5 and ß'-loop-ß'') and the α2 helix to facilitate single-stranded substrate binding. Moreover, a bubble-like region around Asp194 and a key residue inside the active pocket (Thr133) enabled specific recognition and demethylation of m1 A- and 3-methylcytidine (m3 C)-modified substrates. Mutation of Thr133 to the corresponding residue in the AlkB Fe(II)/α-ketoglutarate-dependent dioxygenase family members FTO or ALKBH5 converted ALKBH3 substrate selectivity from m1 A to N6 -methyladenosine (m6 A), as did Asp194 deletion. Our findings provide a molecular basis for understanding the mechanisms of substrate recognition and m1 A demethylation by ALKBH3. This study is expected to aid structure-guided design of chemical probes for further functional studies and therapeutic applications.


Subject(s)
Alpha-Ketoglutarate-Dependent Dioxygenase FTO , RNA , Humans , RNA/chemistry , Alpha-Ketoglutarate-Dependent Dioxygenase FTO/metabolism , RNA, Messenger/metabolism , Demethylation , Ferrous Compounds , AlkB Homolog 3, Alpha-Ketoglutarate-Dependent Dioxygenase/metabolism
5.
Front Plant Sci ; 12: 712713, 2021.
Article in English | MEDLINE | ID: mdl-34386031

ABSTRACT

As the most abundant and reversible chemical modification in eukaryotic mRNA, the epitranscriptomic mark N 6-methyladenine (m6A) regulates plant development and stress response. We have previously characterized that ALKBH10B is an Arabidopsis mRNA m6A demethylase and regulates floral transition. However, it is unclear whether ALKBH10B plays a role in abiotic stress response. Here, we found that the expression of ALKBH10B is increased in response to abscisic acid (ABA), osmotic, and salt stress. The alkbh10b mutants showed hypersensitive to ABA, osmotic, and salt stress during seed germination. Transcriptome analysis revealed that the expression of several ABA response genes is upregulated in alkbh10b-1 than that of wild type, indicating ALKBH10B negatively affects the ABA signaling. Furthermore, m6A sequencing showed that ABA signaling genes, including PYR1, PYL7, PYL9, ABI1, and SnRK2.2 are m6A hypermethylated in alkbh10b-1 after ABA treatment. Taken together, our work demonstrated that ALKBH10B negatively modulates ABA response during seed germination in Arabidopsis.

6.
Nat Biotechnol ; 39(12): 1581-1588, 2021 12.
Article in English | MEDLINE | ID: mdl-34294912

ABSTRACT

RNA N6-methyladenosine (m6A) modifications are essential in plants. Here, we show that transgenic expression of the human RNA demethylase FTO in rice caused a more than threefold increase in grain yield under greenhouse conditions. In field trials, transgenic expression of FTO in rice and potato caused ~50% increases in yield and biomass. We demonstrate that the presence of FTO stimulates root meristem cell proliferation and tiller bud formation and promotes photosynthetic efficiency and drought tolerance but has no effect on mature cell size, shoot meristem cell proliferation, root diameter, plant height or ploidy. FTO mediates substantial m6A demethylation (around 7% of demethylation in poly(A) RNA and around 35% decrease of m6A in non-ribosomal nuclear RNA) in plant RNA, inducing chromatin openness and transcriptional activation. Therefore, modulation of plant RNA m6A methylation is a promising strategy to dramatically improve plant growth and crop yield.


Subject(s)
Oryza , Solanum tuberosum , Alpha-Ketoglutarate-Dependent Dioxygenase FTO/metabolism , Biomass , Demethylation , Humans , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , RNA, Plant/genetics , Solanum tuberosum/genetics
7.
Nat Commun ; 12(1): 1790, 2021 03 19.
Article in English | MEDLINE | ID: mdl-33741984

ABSTRACT

RNA-mediated chromatin silencing is central to genome regulation in many organisms. However, how nascent non-coding transcripts regulate chromatin is poorly understood. Here, through analysis of Arabidopsis FLC, we show that resolution of a nascent-transcript-induced R-loop promotes chromatin silencing. Stabilization of an antisense-induced R-loop at the 3' end of FLC enables an RNA binding protein FCA, with its direct partner FY/WDR33 and other 3'-end processing factors, to polyadenylate the nascent antisense transcript. This clears the R-loop and recruits the chromatin modifiers demethylating H3K4me1. FCA immunoprecipitates with components of the m6A writer complex, and m6A modification affects dynamics of FCA nuclear condensates, and promotes FLC chromatin silencing. This mechanism also targets other loci in the Arabidopsis genome, and consistent with this fca and fy are hypersensitive to a DNA damage-inducing drug. These results show how modulation of R-loop stability by co-transcriptional RNA processing can trigger chromatin silencing.


Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis/genetics , Chromatin/genetics , Flowers/genetics , Gene Silencing , MADS Domain Proteins/genetics , R-Loop Structures , RNA-Binding Proteins/genetics , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Chromatin/metabolism , Flowers/metabolism , Gene Expression Regulation, Plant , MADS Domain Proteins/metabolism , Polyadenylation , Protein Binding , RNA Stability/genetics , RNA, Plant/chemistry , RNA, Plant/genetics , RNA, Plant/metabolism , RNA-Binding Proteins/metabolism , Transcription, Genetic , mRNA Cleavage and Polyadenylation Factors/genetics , mRNA Cleavage and Polyadenylation Factors/metabolism
8.
Wiley Interdiscip Rev RNA ; 10(1): e1507, 2019 01.
Article in English | MEDLINE | ID: mdl-30252201

ABSTRACT

N6 -methyladenosine (m6 A) is the most abundant internal chemical modification in eukaryotic messenger RNAs (mRNAs). The discovery in 2011 that m6 A is reversed by the fat mass and obesity-associated protein stimulated extensive worldwide research efforts on the regulatory biological functions of dynamic m6 A and other RNA modifications. The epitranscriptomic mark m6 A is written, read, and erased through the activities of a complicated network of enzymes and other proteins. m6 A-binding proteins read m6 A marks and transduce their downstream regulatory effects by altering RNA metabolic processes. In this review, we summarize the current knowledge of m6 A modifications, with particular focus on the functions of its writer, eraser, and reader proteins in posttranscriptional gene regulation and discuss the impact of m6 A marks on human health. This article is categorized under: RNA Processing > RNA Editing and Modification RNA in Disease and Development > RNA in Disease.


Subject(s)
Adenosine/analogs & derivatives , RNA/metabolism , Adenosine/metabolism , Animals , Carrier Proteins/metabolism , Histone Demethylases/metabolism , Humans , Methylation , Methyltransferases/metabolism
9.
Plant Cell ; 30(5): 968-985, 2018 05.
Article in English | MEDLINE | ID: mdl-29716990

ABSTRACT

The epitranscriptomic mark N6-methyladenosine (m6A) can be written, read, and erased via the action of a complex network of proteins. m6A binding proteins read m6A marks and transduce their downstream regulatory effects by altering RNA metabolic processes. The characterization of m6A readers is an essential prerequisite for understanding the roles of m6A in plants, but the identities of m6A readers have been unclear. Here, we characterized the YTH-domain family protein ECT2 as an Arabidopsis thaliana m6A reader whose m6A binding function is required for normal trichome morphology. We developed the formaldehyde cross-linking and immunoprecipitation method to identify ECT2-RNA interaction sites at the transcriptome-wide level. This analysis demonstrated that ECT2 binding sites are strongly enriched in the 3' untranslated regions (3' UTRs) of target genes and led to the identification of a plant-specific m6A motif. Sequencing analysis suggested that ECT2 plays dual roles in regulating 3' UTR processing in the nucleus and facilitating mRNA stability in the cytoplasm. Disruption of ECT2 accelerated the degradation of three ECT2 binding transcripts related to trichome morphogenesis, thereby affecting trichome branching. The results shed light on the underlying mechanisms of the roles of m6A in RNA metabolism, as well as plant development and physiology.


Subject(s)
Arabidopsis/metabolism , Trichomes/genetics , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Binding Sites , Intracellular Signaling Peptides and Proteins/genetics , Intracellular Signaling Peptides and Proteins/metabolism , Protein Binding , RNA Stability/genetics , RNA Stability/physiology
10.
Plant Cell ; 29(12): 2995-3011, 2017 12.
Article in English | MEDLINE | ID: mdl-29180595

ABSTRACT

N6-methyladenosine (m6A) is the most abundant, internal, posttranscriptional modification in mRNA among all higher eukaryotes. In mammals, this modification is reversible and plays broad roles in the regulation of mRNA metabolism and processing. Despite its importance, previous studies on the role and mechanism of m6A methylation in Arabidopsis thaliana have been limited. Here, we report that ALKBH10B is a demethylase that oxidatively reverses m6A methylation in mRNA in vitro and in vivo. Depletion of ALKBH10B in the alkbh10b mutant delays flowering and represses vegetative growth. Complementation with wild-type ALKBH10B, but not a catalytically inactive mutant (ALKBH10B H366A/E368A), rescues these effects in alkbh10b-1 mutant plants, suggesting the observed phenotypes are controlled by the catalytic action of ALKBH10B We show that ALKBH10B-mediated mRNA demethylation affects the stability of target transcripts, thereby influencing floral transition. We identified 1190 m6A hypermethylated transcripts in the alkbh10b-1 mutant involved in plant development. The discovery and characterization of the archetypical RNA demethylase in Arabidopsis sheds light on the occurrence and functional role(s) of reversible mRNA methylation in plants and defines the role of m6A RNA modification in Arabidopsis floral transition.


Subject(s)
Adenosine/analogs & derivatives , Arabidopsis Proteins/metabolism , Arabidopsis/enzymology , Arabidopsis/physiology , Flowers/enzymology , Flowers/physiology , Oxidoreductases, N-Demethylating/metabolism , RNA, Plant/metabolism , RNA-Binding Proteins/metabolism , Adenosine/chemistry , Adenosine/metabolism , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis Proteins/genetics , Demethylation , Flowers/genetics , Gene Expression Regulation, Plant , Genes, Plant , Methylation , Mutation/genetics , Oxidoreductases, N-Demethylating/genetics , Protein Stability , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA-Binding Proteins/genetics , Substrate Specificity , Up-Regulation/genetics
11.
Nat Commun ; 5: 5630, 2014 Nov 28.
Article in English | MEDLINE | ID: mdl-25430002

ABSTRACT

Recent discoveries of reversible N(6)-methyladenosine (m(6)A) methylation on messenger RNA (mRNA) and mapping of m(6)A methylomes in mammals and yeast have revealed potential regulatory functions of this RNA modification. In plants, defects in m(6)A methyltransferase cause an embryo-lethal phenotype, suggesting a critical role of m(6)A in plant development. Here, we profile m(6)A transcriptome-wide in two accessions of Arabidopsis thaliana and reveal that m(6)A is a highly conserved modification of mRNA in plants. Distinct from mammals, m(6)A in A. thaliana is enriched not only around the stop codon and within 3'-untranslated regions, but also around the start codon. Gene ontology analysis indicates that the unique distribution pattern of m(6)A in A. thaliana is associated with plant-specific pathways involving the chloroplast. We also discover a positive correlation between m(6)A deposition and mRNA abundance, suggesting a regulatory role of m(6)A in plant gene expression.


Subject(s)
Adenosine/analogs & derivatives , Arabidopsis , Gene Expression Regulation, Plant , RNA, Messenger/metabolism , 3' Untranslated Regions , Adenosine/metabolism , Codon, Initiator/metabolism , Codon, Terminator/metabolism , Gene Expression Profiling , Methyltransferases/genetics , Methyltransferases/metabolism
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