Circular RNAs (circRNAs) arise during pre-mRNA splicing, in which the 3′ and 5′ ends are linked to each other by a covalent bond. Soybean is an ancient tetraploid, which underwent two whole genome duplications. Most of soybean genes are paralogous genes with multiple copies. Although many circRNAs have been identified in animals and plants, little is known about soybean circRNAs, especially about circRNAs derived from paralogous genes. Here, we used deep sequencing technology coupled with RNase R enrichment strategy and bioinformatic approach to uncover circRNAs in soybean. A total of 5,372 circRNAs were identified, approximately 80% of which were paralogous circRNAs generated from paralogous genes. Despite high sequence homology, the paralogous genes could produce different paralogous circRNAs with different expression patterns. Two thousand and one hundred thirty four circRNAs were predicted to be 92 miRNAs target mimicry. CircRNAs and circRNA isoforms exhibited tissue-specific expression patterns in soybean. Based on the function of circRNA-host genes, the soybean circRNAs may participate in many biological processes such as developmental process, multi-organism process, and metabolic process. Our study not only provided a basis for research into the function of circRNAs in soybean but also new insights into the plant circRNA kingdom.

Sucrose non-fermenting-1 (SNF1)-related protein kinase 2s (SnRK2s) that were reported to be involved in the transduction of abscisic acid (ABA) signaling, play important roles in response to biotic and abiotic stresses in plants. Compared to the systemic investigation of SnRK2s in Arabidopsis thaliana and Oryza sativa, little is known regarding SnRK2s in soybean, which is one of the most important oil and protein crops. In the present study, we performed genome-wide identification and characterization of GmSnRK2s in soybean. In summary, 22 GmSnRK2s were identified and clustered into four groups. Phylogenetic analysis indicated the expansion of SnRK2 gene family during the evolution of soybean. Various cis-acting elements such as ABA Response Elements (ABREs) were identified and analyzed in the promoter regions of GmSnRK2s. The results of RNA sequencing (RNA-Seq) data for different soybean tissues showed that GmSnRK2s exhibited spatio-temporally specific expression patterns during soybean growth and development. Certain GmSnRK2s could respond to the treatments including salinity, ABA and strigolactones. Our results provide a foundation for the further elucidation of the function of GmSnRK2 genes in soybean.

Summary Rhg1 (resistance to Heterodera glycines 1) is an important locus that contributes to resistance against soybean cyst nematode (SCN; Heterodera glycines Ichinohe), which is the most economically damaging disease of soybean worldwide. Simultaneous overexpression of three genes encoding a predicted amino acid transporter, an α‐soluble N ‐ethylmaleimide‐sensitive factor attachment protein (α‐SNAP) and a predicted wound‐induced protein resulted in resistance to SCN provided by this locus. However, the roles of two of these genes (excluding α‐ SNAP ) remain unknown. Here, we report the functional characterization of Glyma.18G022400 , a gene at the Rhg1 locus that encodes the predicted amino acid transporter Rhg1‐GmAAT. Although the direct role of Rhg1‐GmAAT in glutamate transport was not demonstrated, multiple lines of evidence showed that Rhg1‐GmAAT impacts glutamic acid tolerance and glutamate transportation in soybean. Transcriptomic and metabolite profiling indicated that overexpression of Rhg1‐GmAAT activated the jasmonic acid (JA) pathway. Treatment with a JA biosynthesis inhibitor reduced the resistance provided by the Rhg1 ‐containing PI88788 to SCN, which suggested that the JA pathway might play a role in Rhg1 ‐mediated resistance to SCN. Our results could be helpful for the clarification of the mechanism of resistance to SCN provided by Rhg1 in soybean.

Objective To investigate the effectiveness of robot-assisted therapy on balance function in stroke survivors. Data sources PubMed, the Cochrane Library, Embase and China National Knowledge Infrastructure databases were searched systematically for relevant studies. Study selection Randomized controlled trials reporting robot-assisted therapy on balance function in patients after stroke were included. Data extraction Information on study characteristics, demographics, interventions strategies and outcome measures were extracted by 2 reviewers. Data synthesis A total of 19 randomized trials fulfilled the inclusion criteria and 13 out of 19 were included in the meta-analysis. Analysis revealed that robot-assisted therapy significantly improved balance function assessed by berg balance scale (weighted mean difference (WMD) 3.58, 95% confidence interval (95% CI) 1.89–5.28, p < 0.001) compared with conventional therapy. Secondary analysis indicated that there was a significant difference in balance recovery between the conventional therapy and robot-assisted therapy groups in the acute/subacute stages of stroke (WMD 5.40, 95% CI 3.94–6.86, p < 0.001), while it was not significant in the chronic stages. With exoskeleton devices, the balance recovery in robot-assisted therapy groups was significantly better than in the conventional therapy groups (WMD 3.73, 95% CI 1.83–5.63, p < 0.001). Analysis further revealed that a total training time of more than 10 h can significantly improve balance function (WMD 4.53, 95% CI 2.31–6.75, p < 0.001). No publication bias or small study effects were observed according to the Cochrane Collaboration tool. Conclusion These results suggest that robot-assisted therapy is an effective intervention for improving balance function in stroke survivors. LAY ABSTRACT Balance is an important factor in ability to perform independent walking. Many patients with stroke gain little benefit from neural rehabilitation because their balance control is impaired. Robot-assisted therapy is a promising intervention approach, which has developed rapidly in recent years. Several previous reviews have focused on gait-related measurements, such as walking speed and endurance; however, the effectiveness of robot-assisted therapy on balance has not been clearly outlined. This systematic review and meta-analysis showed that robot-assisted therapy can significantly improve balance recovery compared with conventional therapy, especially for people in the acute/subacute phase after stroke treated with an exoskeleton and a total training time of more than 10 h.

Objectives: The aim of this study was to investigate the effects of inspiratory muscle training in post-stroke patients and to explore the effective training protocol. Data sources: PubMed/Medline, Web of Science, Scopus, Embase, Cochrane database, China National Knowledge Infrastructure, and China Science Periodical Database were searched through April 2020. Review methods: Trials examining effects of inspiratory muscle training on pulmonary function, cardiopulmonary endurance, pulmonary infection incidence, and quality of life in post-stroke patients were included. Subgroup analysis was performed to compare different training programs. Mean differences and risk ratios with 95% confidence intervals were presented. Risk of bias was assessed with the Cochrane tool. Results: Thirteen randomized controlled trials involving a total of 373 participants were identified. Meta-analysis conducted in 8 out of 13 trials revealed evidence for beneficial effects of inspiratory muscle training on forced vital capacity (MD: 0.47, 95% CI: 0.28–0.66), forced expired volume in 1 second (MD: 0.26, 95% CI: 0.18–0.35), 6-minute walk test (MD: 52.61, 95% CI: 25.22–80.01), maximum inspiratory pressure (MD: 18.18, 95% CI: 5.58–30.78), inspiratory muscle endurance (MD: 19.99, 95% CI: 13.58–26.40), and pulmonary infection incidence (RR: 0.11, 95% CI: 0.03–0.40). Omitting individual trials from the meta-analysis did not significantly change the results. The effective inspiratory muscle training protocol was suggested by subgroup analysis with three repetitions per week and more than 20 minutes per day for three weeks. Conclusion: Inspiratory muscle training can be considered as an effective intervention for improving pulmonary function and cardiopulmonary endurance, and reducing pulmonary infection incidence in patients after stroke.

Background: Lung cancer surgery is associated with a high incidence of postoperative pulmonary complications (PPCs). Preliminary evidence suggests that ERAS processes can reduce overall incidence of PPCs as short-and longterm recovery improved by supporting units to adopt evidence-based care. However, the evidence is inconclusive due to insufficient high-level studies in this research field. No well-designed, adequately powered, randomized controlled trials (RCTs) have investigated the effects of pulmonary rehabilitation based ERAS program (PREP) on post-operative pulmonary complications, pulmonary function, and health related quality of life following lung cancer surgery. Methods:The PREP trial is a pragmatic, investigator-initiated, multi-center, randomized controlled, parallel group, clinical trial. Five hundred patients scheduled for minimally invasive pulmonary resection at six hospitals in China will be randomized with concealed allocation to receive either i) a pre-operative assessment and an information booklet or ii) a pre-operative assessment, an information booklet, plus an additional education, a 30-min pulmonary rehabilitation training session and the post-operative pulmonary rehabilitation program. The primary outcome is incidence of PPCs defined with the Melbourne Group Scale diagnostic scoring tool. Secondary outcomes include incidence of cardiopulmonary and other complications, pulmonary function, cardiopulmonary endurance, muscle strength, activity level, health-related quality of life (HRQoL), pre-and post-operative hospital length of stay (LOS), and total hospital LOS. Discussion: The PREP trial is designed to verify the hypothesis that pulmonary rehabilitation based ERAS program reduces incidence of PPCs and improves pulmonary function and HRQoL in patients following lung cancer surgery. This trial will furthermore contribute significantly to the limited knowledge about the pulmonary rehabilitation based ERAS program following lung cancer surgery, and may thereby form the basis of future recommendations in the surgical community.

Objective: Increasing evidence has uncovered the roles of lncRNA-miRNA-mRNA regulatory networks in cardiovascular diseases. However, the crosstalk between ceRNA networks and development of heart failure (HF) remains unclear. This study was to investigate the role of lncRNA-mediated ceRNA networks in the pathophysiological process of HF and its potential regulatory functions on programmed cell death.Methods: We firstly screened the GSE77399, GSE52601 and GSE57338 datasets in the NCBI GEO database for screening differentially expressed lncRNAs, miRNAs and mRNAs. lncRNA-miRNA-mRNA regulatory networks based on the ceRNA theory were subsequently constructed. GO and KEGG enrichment analysis was conducted to predict potential biological functions of mRNAs in ceRNA networks. Differentially expressed mRNAs were then interacted with programmed cell death related genes. lncRNA-mediated ceRNA regulatory pathways on programmed cell death were validated with qRT-PCR testing.Results: Based on our bioinformatic analysis, two lncRNAs, eight miRNAs and 65 mRNAs were extracted to construct two lncRNAs-mediated ceRNA networks in HF. Biological processes and pathways were enriched in extracellular matrix. Seven lncRNA-mediated ceRNA regulatory pathways on programmed cell death, GAS5/miR-345-5p/ADAMTS4, GAS5/miR-18b-5p/AQP3, GAS5/miR-18b-5p/SHISA3, GAS5/miR-18b-5p/C1orf105, GAS5/miR-18b-5p/PLIN2, GAS5/miR-185-5p/LPCAT3, and GAS5/miR-29b-3p/STAT3, were finally validated.Conclusions: Two novel ceRNA regulatory networks in HF were discovered based on our bioinformatic analysis. Based on the interaction and validation analysis, seven lncRNA GAS5-mediated ceRNA regulatory pathways were hypothesized to impact programmed cell death including seven for apoptosis, three for ferroptosis, and one for pyroptosis. Upon which, we provided novel insights and potential research plots for bridging ceRNA regulatory networks and programmed cell death in HF.

Molecular breeding has become an important approach for crop improvement, and a prerequisite for molecular breeding is elucidation of the functions of genetic loci or genes. Soybean is one of the most important food and oil crops worldwide. However, due to the difficulty of genetic transformation in soybean, studies of its functional genomics lag far behind those of other crops such as rice, which severely impairs the progress of molecular improvement in soybean. Here, we describe an effective large-scale strategy to investigate the functions of soybean genes via overexpression of a full-length soybean cDNA library in Arabidopsis. The overexpression vector pJL12 was modified for use in the construction of a normalized full-length cDNA library. The constructed cDNA library showed good quality; repetitive clones represented approximately 4%, insertion fragments were approximately 2.2 kb, and the full-length rate was approximately 98%. This cDNA library was then overexpressed in Arabidopsis, and approximately 2000 transgenic lines were preliminarily obtained. Phenotypic analyses of the positive T1 transgenic plants showed that more than 5% of the T1 transgenic lines displayed abnormal developmental phenotypes, and approximately 1% of the transgenic lines exhibited potentially favorable traits. We randomly amplified 4 genes with obvious phenotypes (enlarged seeds, yellowish leaves, more branches, and dense siliques) and repeated the transgenic analyses in Arabidopsis. Subsequent phenotypic observation demonstrated that these phenotypes were indeed due to the overexpression of soybean genes. We believe our strategy represents an effective large-scale approach to investigate the functions of soybean genes and further reveal genes favorable for molecular improvement in soybean.