The Myb73-GDPD2-GA2ox1 transcriptional regulatory module confers phosphate deficiency tolerance in soybean.

Phosphorus is indispensable in agricultural production. An increasing food supply requires more efficient use of phosphate due to limited phosphate resources. However, how crops regulate phosphate efficiency remains largely unknown. Here, we identified a major quantitative trait locus, qPE19, that controls 7 low-phosphate (LP)-related traits in soybean (Glycine max) through linkage mapping and genome-wide association studies. We identified the gene responsible for qPE19 as GLYCEROPHOSPHORYL DIESTER PHOSPHODIESTERASE2 (GmGDPD2), and haplotype 5 represents the optimal allele favoring LP tolerance. Overexpression of GmGDPD2 significantly affects hormone signaling and improves root architecture, phosphate efficiency and yield-related traits; conversely, CRISPR/Cas9-edited plants show decreases in these traits. GmMyb73 negatively regulates GmGDPD2 by directly binding to its promoter; thus, GmMyb73 negatively regulates LP tolerance. GmGDPD2 physically interacts with GA 2-oxidase 1 (GmGA2ox1) in the plasma membrane, and overexpressing GmGA2ox1 enhances LP-associated traits, similar to GmGDPD2 overexpression. Analysis of double mutants for GmGDPD2 and GmGA2ox1 demonstrated that GmGDPD2 regulates LP tolerance likely by influencing auxin and gibberellin dose-associated cell division in the root. These results reveal a regulatory module that plays a major role in regulating LP tolerance in soybeans and is expected to be utilized to develop phosphate-efficient varieties to enhance soybean production, particularly in phosphate-deficient soils.

GRN mutation spectrum and genotype-phenotype correlation in Chinese dementia patients: data from PUMCH dementia cohort.

BACKGROUND: METHODS: The GRN mutations, especially of the loss of function type, are causative of frontotemporal dementia (FTD). However, several GRN variants can be found in other neurodegenerative diseases, such as Alzheimer's disease (AD) and Parkinson's disease. So far, there have been over 300 GRN mutations reported globally. However, the genetic spectrum and phenotypic characteristics have not been fully elucidated in Chinese population.The participants were from the dementia cohort of Peking Union Medical College Hospital (n=1945). They received history inquiry, cognitive evaluation, brain imaging and exome sequencing. The dementia subjects carrying the rare variants of the GRN were included in this study. Those with the pathogenic or likely pathogenic variants of other dementia-related genes were excluded. RESULTS: 14 subjects carried the rare variants of GRN. They were clinically diagnosed with behavioural variant of FTD (n=2), non-fluent/agrammatic variant primary progressive aphasia (PPA, n=3), semantic variant PPA (n=1), AD (n=6) and mixed dementia (n=2). 13 rare variants of GRN were found, including 6 novel variants (W49X, S226G, M152I, A91E, G79E and A303S). The most prevalent symptom was amnesia (85.7%, 12/14), followed by psychiatric and behavioural disorder (78.6%, 11/14). In terms of lobar atrophy, temporal atrophy/hypometabolism was the most common (85.7%, 12/14), followed by parietal atrophy/hypometabolism (78.6%, 11/14). CONCLUSION: The novel GRN variants identified in this study contribute to enrich the GRN mutation repertoire. There is phenotypic similarity and diversity among Chinese patients with the GRN mutations.

WGCNA and transcriptome profiling reveal hub genes for key development stage seed size/oil content between wild and cultivated soybean.

BACKGROUND: Soybean is one of the most important oil crops in the world. The domestication of wild soybean has resulted in significant changes in the seed oil content and seed size of cultivated soybeans. To better understand the molecular mechanisms of seed formation and oil content accumulation, WDD01514 (E1), ZYD00463 (E2), and two extreme progenies (E23 and E171) derived from RILs were used for weighted gene coexpression network analysis (WGCNA) combined with transcriptome analysis. RESULTS: In this study, both seed weight and oil content in E1 and E171 were significantly higher than those in E2 and E23, and 20 DAF and 30 DAF may be key stages of soybean seed oil content accumulation and weight increase. Pathways such as "Photosynthesis", "Carbon metabolism", and "Fatty acid metabolism", were involved in oil content accumulation and grain formation between wild and cultivated soybeans at 20 and 30 DAF according to RNA-seq analysis. A total of 121 oil content accumulation and 189 seed formation candidate genes were screened from differentially expressed genes. WGCNA identified six modules related to seed oil content and seed weight, and 76 candidate genes were screened from modules and network. Among them, 16 genes were used for qRT-PCR and tissue specific expression pattern analysis, and their expression-levels in 33-wild and 23-cultivated soybean varieties were subjected to correlation analysis; some key genes were verified as likely to be involved in oil content accumulation and grain formation. CONCLUSIONS: Overall, these results contribute to an understanding of seed lipid metabolism and seed size during seed development, and identify potential functional genes for improving soybean yield and seed oil quantity.

GmEIL4 enhances soybean (Glycine max) phosphorus efficiency by improving root system development.

Phosphorus (P) deficiency seriously affects plant growth and development and ultimately limits the quality and yield of crops. Here, a new P efficiency-related major quantitative trait locus gene, GmEIL4 (encoding an ethylene-insensitive 3-like 1 protein), was cloned at qP2, which was identified by linkage analysis and genome-wide association study across four environments. Overexpressing GmEIL4 significantly improved the P uptake efficiency by increasing the number, length and surface area of lateral roots of hairy roots in transgenic soybeans, while interfering with GmEIL4 resulted in poor root phenotypic characteristics compared with the control plants under low P conditions. Interestingly, we found that GmEIL4 interacted with EIN3-binding F box protein 1 (GmEBF1), which may regulate the root response to low P stress. We conclude that the expression of GmEIL4 was induced by low-P stress and that overexpressing GmEIL4 improved P accumulation by regulating root elongation and architecture. Analysis of allele variation of GmEIL4 in 894 soybean accessions suggested that GmEIL4 is undergoing artificial selection during soybean evolution, which will benefit soybean production. Together, this study further elucidates how plants respond to low P stress by modifying root structure and provides insight into the great potential of GmEIL4 in crop P-efficient breeding.

Clinical-neuroimaging-pathological relationship analysis of adult onset Neuronal Intranuclear Inclusion Disease (NIID).

BACKGROUND: Neuronal Intranuclear Inclusion Disease (NIID) is a degenerative disease with heterogeneous clinical manifestations. We aim to analysis the relationship between clinical manifestations, neuroimaging and skin pathology in a Chinese NIID cohort. METHODS: Patients were recruited from a Chinese cohort. Detail clinical information were collected. Visual rating scale was used for evaluation of neuroimaging. The relationship between clinical presentations and neuroimaging, as well as skin pathology was statistically analyzed. RESULTS: Thirty-two patients were recruited. The average onset age was 54.3 y/o. 28.1% had positive family history. Dementia, autonomic nervous system dysfunction, episodic attacks were three main presentations. CSF analysis including Aß42 and tau level was almost normal. The most frequently involved on MRI was periventricular white matter (100%), frontal subcortical and deep white matter (96.6%), corpus callosum (93.1%) and external capsule (72.4%). Corticomedullary junction DWI high intensity was found in 87.1% patients. Frontal and external capsule DWI high intensity connected to form a "kite-like" specific image. Severity of dementia was significantly related to leukoencephalopathy (r = 0.465, p = 0.0254), but not cortical atrophy and ventricular enlargement. Grey matter lesions were significantly associated with encephalopathy like attacks (p = 0.00077) but not stroke like attacks. The density of intranuclear inclusions in skin biopsy was not associated with disease duration, severity of leukoencephalopathy and dementia. CONCLUSIONS: Specific distribution of leukoencephalopathy and DWI high intensity were indicative. Leukoencephalopathy and subcortical mechanism were critical in pathogenesis of NIID. Irrelevant of inclusion density and clinical map suggested the direct pathogenic factor need further investigation.

Typical and atypical phenotype and neuroimaging of X-linked adrenoleukodystrophy in a Chinese cohort.

OBJECTIVE: The objective of this study is to describe the typical and atypical clinical and neuroimaging features of ALD in Chinese patients, which will help early diagnosis and intervention to improve prognosis of ALD. METHODS: Forty-one patients in the Leukoencephalopathy Clinic of Neurology Department, Peking Union Medical College Hospital were enrolled. Detailed clinical manifestations and MRI features were analyzed. The relationship between phenotype and genotype as well as biochemical analysis was observed. RESULTS: The patients were classified according to phenotype and onset age, including 14 childhood cerebral ALD (CCALD), 8 adolescent cerebral ALD (adoCALD), 3 adult cerebral ALD (ACALD), 14 adrenomyeloneuropathy (AMN), and 2 ALD in women. AMN was the main presentation in adults. Visual impairment was usual onset symptom in CCALD and cognitive decline and psychiatric symptoms were found in adoCALD and ACALD. Typical MRI feature of CALD was symmetrical peri-ventricular "butterfly wings" like lesions in frontal and/or occipital lobe with peripheral DWI hyperintensities and Gd enhancement. Corpus callosum and internal capsule were always involved. Unilateral lesions were also possible. Cerebral AMN presented with centrum semiovale diffuse involvement. Spinocerebellar variant was a special subtype of AMN with obvious cerebellar and brainstem lesions. No relationships between phenotype and genotype as well as biochemical VLCFAs analysis were found. CONCLUSIONS: We emphasize that corpus callosum and internal capsule are always involved in ALD. A unilateral lesion is also possible. Neuroimaging of cerebral AMN is different from typical CALD with more centrum semiovale involvement. We support spinocerebellar variant was a rare subtype of AMN.

Comparative analysis in different organs and tissue-specific metabolite profiling of Atractylodes lancea from four regions by GC-MS and laser microdissection.

Traditional Chinese medicine is made from the rhizome of Atractylodes lancea (Thunb.) DC. (Compositae), known as Cangzhu. In this study, gas chromatography-mass spectrometry was used to identify and quantify the volatile oils of different organs of A. lancea from four regions of China: Jiangsu, Anhui, Henan, and Hubei provinces. The volatile oils of A. lancea were qualitatively and quantitatively characterized using gas chromatography-mass spectrometry combined with laser microdissection. The results identified 21 components in A. lancea, the majority of the components were found in the rhizomes, followed by the fibrous roots, flowers, leaves, and stems. According to the contents of volatile oils in A. lancea, it was divided into Dabieshan (mainly includes hinesol and ß-eudesmol) and Maoshan types (mainly includes atractylon and atractylodin), and the ratios of hinesol:ß-eudesmol:atractylon:atractylodin were 17.06:4.55:0:1, 12.66:11.71:0.99:1, 7.43:6.23:0:1, and 0.13:0.16:1.52:1 in A. lancea from AH, HN, HB, and JS, respectively. Tissue-specific study indicated that Dabieshan type mainly includes elemol, hinesol, and ß-eudesmol in the periderm and secretory cavities of A. lancea, whereas Maoshan type mainly includes atractylon, atractylodin, little hinesol, and ß-eudesmol in the secretory cavities. Conversely, no volatile oils were detected in the cortex, phloem, xylem, vascular ray, or pith. This study provides a foundation for further evaluation and utilization of A. lancea.

Artificial selection on GmOLEO1 contributes to the increase in seed oil during soybean domestication.

Increasing seed oil content is one of the most important breeding goals for soybean due to a high global demand for edible vegetable oil. However, genetic improvement of seed oil content has been difficult in soybean because of the complexity of oil metabolism. Determining the major variants and molecular mechanisms conferring oil accumulation is critical for substantial oil enhancement in soybean and other oilseed crops. In this study, we evaluated the seed oil contents of 219 diverse soybean accessions across six different environments and dissected the underlying mechanism using a high-resolution genome-wide association study (GWAS). An environmentally stable quantitative trait locus (QTL), GqOil20, significantly associated with oil content was identified, accounting for 23.70% of the total phenotypic variance of seed oil across multiple environments. Haplotype and expression analyses indicate that an oleosin protein-encoding gene (GmOLEO1), colocated with a leading single nucleotide polymorphism (SNP) from the GWAS, was significantly correlated with seed oil content. GmOLEO1 is predominantly expressed during seed maturation, and GmOLEO1 is localized to accumulated oil bodies (OBs) in maturing seeds. Overexpression of GmOLEO1 significantly enriched smaller OBs and increased seed oil content by 10.6% compared with those of control seeds. A time-course transcriptomics analysis between transgenic and control soybeans indicated that GmOLEO1 positively enhanced oil accumulation by affecting triacylglycerol metabolism. Our results also showed that strong artificial selection had occurred in the promoter region of GmOLEO1, which resulted in its high expression in cultivated soybean relative to wild soybean, leading to increased seed oil accumulation. The GmOLEO1 locus may serve as a direct target for both genetic engineering and selection for soybean oil improvement.

Colour, chemical compounds, and antioxidant capacity of Astragali Radix based on untargeted metabolomics and targeted quantification.

INTRODUCTION: Astragali Radix has been used for over 2000 years in traditional Chinese medicine. Its secondary xylem "Jinjing" and secondary phloem "Yulan" are important for evaluating the quality of the Daodi medicinal material in China. However, its systematic characterisation has not been conducted. OBJECTIVE: This study aims to investigate the colour, chemical compounds, and antioxidant capacity of the secondary xylem and phloem of Astragali Radix on the basis of untargeted metabolomics, broadening the application scope of Astragali Radix in food and pharmaceutical industries. METHODS: The L*, a*, and b* of the secondary xylem and phloem were measured by colorimetry, and the chemical compounds were identified and quantified by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and high-performance liquid chromatography-diode array detector-evaporative light scattering detection. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays were conducted to evaluate their antioxidant capacity. RESULTS: Thirty-one compounds were identified by UPLC-Q-TOF-MS. The secondary xylem exhibited high parameter b*, flavonoid content, and antioxidant capacity, while the secondary phloem was rich in astragalosides. The colour parameters of well-defined type A significantly varied from those of the other types. Well-defined type A also exhibited the highest antioxidant activity and flavonoid content, followed by middle type A-like, middle type B-like, and yellow shading type B. CONCLUSION: The colour parameters, chemical compounds, and antioxidant capacity among the different transverse sections of secondary xylem and phloem varied. The yellow colour of secondary xylem was correlated to high flavonoid content and antioxidant activity, and well-defined type A of Astragali Radix had better quality than other types.

Outcomes of hip arthroplasty in Parkinson's disease: a meta-analysis and systematic review.

PURPOSE: Hip arthroplasty in Parkinson's disease (PD) is considered to be associated with higher mortality, perioperative complications, and worse clinical outcomes. The purpose of this study was to evaluate the outcomes of hip arthroplasty in PD. METHODS: The PUBMED, EMBASE, and Cochrane Library databases were searched for all available studies comparing the outcomes of hip arthroplasty in PD and non-PD patients. The quality of the studies was scored using the Newcastle-Ottawa scale (NOS). Meta-analysis was performed using Stata 12.0 software. RESULTS: Six retrospective cohort studies with a total of 59,878 patients were included. Among them, there were 15,313 patients in the Parkinson's group and 44,565 in the non-Parkinson's group. Results showed that patients in the Parkinson's group were more likely to have periprosthetic fracture (OR = 7.93, 95% CI, 1.96-32.04; P = 0.004), revision surgery (OR = 2.33, 95% CI, 1.37-3.98; P = 0.002), deep vein thrombosis (OR = 2.34, 95% CI, 1.56-3.49; P < 0.01), and urinary tract infection (OR = 1.34, 95% CI, 1.20-1.49; P < 0.01) than those in the non-Parkinson's group. The Parkinson's group had a longer hospital stay (WMD = 0.38, 95% CI, 0.24-0.52; P < 0.01) and higher hospitalization costs (WMD = 2487.08, 95% CI, 225.99-4748.17; P = 0.03), but there was no significant difference in short-term postoperative mortality (OR = 1.07, 95% CI, 0.65-1.73; P = 0.80). Hip arthroplasty can significantly improve the function of patients with PD and reduce pain. CONCLUSION: Although hip arthroplasty in patients with PD is associated with a higher revision rate and potential risk, it does not increase short-term mortality and can reduce pain, improve function, and improve quality of life. The periprosthetic fracture risk and revision rate are significantly higher and should be accounted for when deciding on operative treatment and communicated to patient with PD.

Validation of the predictive model for seizure recurrence after withdrawal of antiepileptic drugs.

PURPOSE: The purpose of this study was to validate the practicability of Lamberink's prediction model in risk assessment of antiepileptic drug (AED) withdrawal in a real, seizure-free population and to find a practical cutoff value to guide clinical withdrawal. METHODS: A group of seizure-free patients from West China Hospital was recruited. Each patient had been seizure-free for at least two years. The seizure recurrence risk among the patients was calculated by an online AED withdrawal risk calculator. The predictive ability of Lamberink's model was assessed by analyzing discrimination and calibration with receiver operating characteristic (ROC) curves and calibration plots, respectively. RESULTS: A total of 184 seizure-free patients received risk evaluation, all of whom were followed up for at least two years or had an earlier report of seizure relapse. Of these patients, 128 patients were followed up for at least five years or had an earlier report of relapse within five years. Sixty-two of 184 (33.7%) patients relapsed within two years, while 81 of 184 (44.0%) patients relapsed within five years after the start of AEDs' withdrawal. Cox regression analyses showed that seizure duration before remission and the age of seizure onset were independent predictors of relapse at two years. For predictors of recurrence at five years, seizure duration before remission, age at onset, and withdrawal were significant. For discrimination, ROC curve analysis showed that the area under the curve (AUC) for the seizure recurrence within two and five years was 0.605 (95% confidence interval [CI]: 0.518-0.692, p = 0.02) and 0.656 (95% CI: 0.563-0.749, p = 0.003), respectively. For calibration, it was poor in two-year prediction; the observed number was considerably lower than the predicted number. However, the calibration plot showed good calibration with the five-year prediction except for the second, fourth, and eighth deciles. With a cutoff two-year recurrence risk of 47%, the model had a sensitivity of 0.758 and a specificity of 0.410; the largest Youden index was 1.168. With a cutoff five-year recurrence risk of 77%, the model had a sensitivity of 0.358 and a specificity of 0.979; the largest Youden index was 1.337. CONCLUSIONS: Lamberink's prediction model has a general discrimination ability. The model overestimated the actual recurrence events when predicting the two-year recurrence risk, but it showed relatively good calibration with five-year prediction. The cutoff value found in this study may be used to guide patients and clinicians towards a decision regarding the withdrawal of AEDs. The model appears to be a useful tool for predicting seizure recurrence for the five-year recurrence risk.

The impact of diabetes mellitus on carotid artery stenting: a meta-analysis.

Diabetes is considered to be one of the important factors affecting the prognosis of patients undergoing carotid endarterectomy (CEA), but its effect on carotid stenting (CAS) has not been determined. We performed this analysis to evaluate the impact of diabetes mellitus (DM) on carotid stenting. Studies were searched in the MEDLINE, EMBASE, and Cochrane Library databases for all available studies comparing the outcomes of diabetic and nondiabetic patients who underwent CAS. A meta-analysis was performed using the Mantel-Haenszel model to calculate odds ratios (ORs) with RevMan 5.3 software. Five studies with high quality were ultimately included. A total of 3364 patients underwent CAS, including 1042 diabetic patients and 2322 nondiabetic patients. Meta-analysis showed that there was no statistically significant difference in the risk of perioperative stroke(OR,1.20;95%CI,0.74-1.97;P = 0.46;I2 = 24%), transient ischemic attack(TIA)(OR,1.46;95%CI,0.80-2.66;P=0.22;I2=37%), myocardial infarction(MI)(OR,1.12;95%CI,0.47-2.67;P=0.80;I2=0%), mortality (OR,1.77;95%CI,0.91-3.42;P = 0.09;I2=0%), and composite of stroke or death (OR,1.26;95%CI,0.87-1.81;P = 0.22;I2=35%) between diabetic and nondiabetic patients receiving CAS. Sensitivity analysis showed that the meta-analysis results of all outcome events were stable. Compared with nondiabetic patients, diabetes did not increase the risk of the following: perioperative stroke, TIA, MI, mortality, and composite of stroke or death in patients receiving CAS. This study may be used to guide carotid artery stenosis patients with diabetes to choose the optimal treatment option. However, due to the limited number of studies and the different technical levels of institutions, the results should be interpreted caution.

An Artificial Neural Network Prediction Model for Posttraumatic Epilepsy: Retrospective Cohort Study.

BACKGROUND: Posttraumatic epilepsy (PTE) is a common sequela after traumatic brain injury (TBI), and identifying high-risk patients with PTE is necessary for their better treatment. Although artificial neural network (ANN) prediction models have been reported and are superior to traditional models, the ANN prediction model for PTE is lacking. OBJECTIVE: We aim to train and validate an ANN model to anticipate the risks of PTE. METHODS: The training cohort was TBI patients registered at West China Hospital. We used a 5-fold cross-validation approach to train and test the ANN model to avoid overfitting; 21 independent variables were used as input neurons in the ANN models, using a back-propagation algorithm to minimize the loss function. Finally, we obtained sensitivity, specificity, and accuracy of each ANN model from the 5 rounds of cross-validation and compared the accuracy with a nomogram prediction model built in our previous work based on the same population. In addition, we evaluated the performance of the model using patients registered at Chengdu Shang Jin Nan Fu Hospital (testing cohort 1) and Sichuan Provincial People's Hospital (testing cohort 2) between January 1, 2013, and March 1, 2015. RESULTS: For the training cohort, we enrolled 1301 TBI patients from January 1, 2011, to December 31, 2017. The prevalence of PTE was 12.8% (166/1301, 95% CI 10.9%-14.6%). Of the TBI patients registered in testing cohort 1, PTE prevalence was 10.5% (44/421, 95% CI 7.5%-13.4%). Of the TBI patients registered in testing cohort 2, PTE prevalence was 6.1% (25/413, 95% CI 3.7%-8.4%). The results of the ANN model show that, the area under the receiver operating characteristic curve in the training cohort was 0.907 (95% CI 0.889-0.924), testing cohort 1 was 0.867 (95% CI 0.842-0.893), and testing cohort 2 was 0.859 (95% CI 0.826-0.890). Second, the average accuracy of the training cohort was 0.557 (95% CI 0.510-0.620), with 0.470 (95% CI 0.414-0.526) in testing cohort 1 and 0.344 (95% CI 0.287-0.401) in testing cohort 2. In addition, sensitivity, specificity, positive predictive values and negative predictors in the training cohort (testing cohort 1 and testing cohort 2) were 0.80 (0.83 and 0.80), 0.86 (0.80 and 0.84), 91% (85% and 78%), and 86% (80% and 83%), respectively. When calibrating this ANN model, Brier scored 0.121 in testing cohort 1 and 0.127 in testing cohort 2. Compared with the nomogram model, the ANN prediction model had a higher accuracy (P=.01). CONCLUSIONS: This study shows that the ANN model can predict the risk of PTE and is superior to the risk estimated based on traditional statistical methods. However, the calibration of the model is a bit poor, and we need to calibrate it on a large sample size set and further improve the model.

[Formation history of Dao-di herbs in Dabie Mountains].

Dabie Mountains, a unique transition area of subtropical and warm temperate zone, span Anhui, Hubei and Henan pro-vinces with a humid and suitable climate. It is rich in traditional Chinese medicine resources including many Dao-di herbs, and has a profound culture of traditional Chinese medicine with many herbalists in the past. This paper combed the historical geography of Dabie Mountains and development vein of medicinal materials recorded in history, and a textual research on the historical evolution of Dao-di herbs and special herbs in Dabie Mountains was carried out. The administrative region of Dabie Mountains has been changing constantly in history, which includes 25 counties and districts of six cities in Anhui, Henan and Hubei provinces at present. Dabie Mountains abound in a variety of Chinese herbal medicines, which have been recorded in herbal works in the past dynasties and local chronicles in detail. The recorded Dao-di herbs Dendrobium huoshanense, Artemisia argyi, and Poria cocos are well-known in China, and the output of Gastrodia elata, Ganoderma lucidum, and P. cocos is in the forefront of the country. Additionally, there are still many local special herbs emerging in modern times. In a word, this paper reviewed development of historical geography and ancient records of medicinal materials in Dabie Mountains, and made a textual research on the Dao-di herbs and special herbs, will provide a reference for the mo-dern research and intellectual property protection of Dao-di herbs in Dabie Mountains.

Genome-wide identification of low phosphorus responsive microRNAs in two soybean genotypes by high-throughput sequencing.

MicroRNAs (miRNAs) have been reported to be correlated with various stress responses in soybean, but only a few miRNAs have been demonstrated to respond to low phosphorus (LP) stress. To unravel the response mechanisms of miRNAs to low-P stress, the roots of two representative soybean genotypes with different P efficiency, Nannong94-156 (a LP-tolerant genotype) and Bogao (a LP-sensitive genotype), were used for the construction of RNA sequencing (RNA-seq) libraries under low/normal-P treatment by high-throughput sequencing. In total, 603 existing miRNAs and 1699 novel miRNAs belonging to 248 and 1582 families in all samples were identified, respectively. Among these miRNAs, 777 miRNAs were differentially expressed (DE) across different P levels and genotypes. Furthermore, putative targets of DE miRNAs were predicted, and these miRNAs mainly targeted ERF (ethylene responsive factor), auxin response factors (ARF), zinc finger protein, MYB, and NAC domain transcription factors. Gene ontology (GO) analysis showed that targets of DE miRNAs were significantly enriched in binding, metabolic processes, biological regulation, response to stress, and phosphorus metabolic processes. In addition, the expression profiles of chosen P-responsive miRNAs and target genes were validated by quantitative real-time PCR (qRT-PCR). Our study focused on genome-wide miRNA identification in two representative soybean genotypes under low-P stress. Overall, the DE miRNAs across different P levels and genotypes and their putative target genes will provide useful information for further study of miRNAs mediating low-P response and facilitate improvements in soybean breeding.

Quantitative trait loci analysis of seed oil content and composition of wild and cultivated soybean.

BACKGROUND: Soybean oil is a major source of edible oil, and the domestication of wild soybean has resulted in significant changes in oil content and composition. Extensive efforts have been made to identify genetic loci that are related to soybean oil traits. The objective of this study was to identify quantitative trait loci (QTLs) related to soybean seed oil and compare the fatty acid composition between wild and cultivated soybean. RESULTS: Using the specific-locus amplified fragment sequencing (SLAF-seq) method, a total of 181 recombinant inbred lines (RILs) derived from a cross between wild soybean ZYD00463 (Glycine soja) and cultivated soybean WDD01514 (Glycine max) were genotyped. Finally, a high-density genetic linkage map comprising 11,398 single-nucleotide polymorphism (SNP) markers on 20 linkage groups (LGs) was constructed. Twenty-four stable QTLs for seed oil content and composition were identified by model-based composite interval mapping (CIM) across multiple environments. Among these QTLs, 23 overlapped with or were adjacent to previously reported QTLs. One QTL, qPA10_1 (5.94-9.98 Mb) on Chr. Ten is a novel locus for palmitic acid. In the intervals of stable QTLs, some interesting genes involved in lipid metabolism were detected. CONCLUSIONS: We developed 181 RILs from a cross between wild soybean ZYD00463 and cultivated soybean WDD01514 and constructed a high-density genetic map using the SLAF-seq method. We identified 24 stable QTLs for seed oil content and compositions, which includes qPA10_1 on Chr. 10, a novel locus for palmitic acid. Some interesting genes in the QTL regions were also detected. Our study will provide useful information for scientists to learn about genetic variations in lipid metabolism between wild and cultivated soybean.

An R2R3-type MYB transcription factor, GmMYB29, regulates isoflavone biosynthesis in soybean.

Isoflavones comprise a group of secondary metabolites produced almost exclusively by plants in the legume family, including soybean [Glycine max (L.) Merr.]. They play vital roles in plant defense and have many beneficial effects on human health. Isoflavone content is a complex quantitative trait controlled by multiple genes, and the genetic mechanisms underlying isoflavone biosynthesis remain largely unknown. Via a genome-wide association study (GWAS), we identified 28 single nucleotide polymorphisms (SNPs) that are significantly associated with isoflavone concentrations in soybean. One of these 28 SNPs was located in the 5'-untranslated region (5'-UTR) of an R2R3-type MYB transcription factor, GmMYB29, and this gene was thus selected as a candidate gene for further analyses. A subcellular localization study confirmed that GmMYB29 was located in the nucleus. Transient reporter gene assays demonstrated that GmMYB29 activated the IFS2 (isoflavone synthase 2) and CHS8 (chalcone synthase 8) gene promoters. Overexpression and RNAi-mediated silencing of GmMYB29 in soybean hairy roots resulted in increased and decreased isoflavone content, respectively. Moreover, a candidate-gene association analysis revealed that 11 natural GmMYB29 polymorphisms were significantly associated with isoflavone contents, and regulation of GmMYB29 expression could partially contribute to the observed phenotypic variation. Taken together, these results provide important genetic insights into the molecular mechanisms underlying isoflavone biosynthesis in soybean.

Cerebrospinal Fluid Alzheimer's Biomarkers and Neurofilament Light Profile of Idiopathic Normal Pressure Hydrocephalus in China: A PUMCH Cohort Study.

INTRODUCTION: Idiopathic normal pressure hydrocephalus (iNPH) is one of the potentially reversible dementias. Early and accurate diagnosis is important for patients' prognosis. Emerging evidence shows fluid biomarkers are useful in diagnosis and pathophysiological research of iNPH. METHODS: Probable iNPH and Alzheimer's disease (AD) patients were recruited. Clinical diagnosis was performed according to international guidelines. CSF collection complied with a standard protocol. Commercial accessible ELISA kits were introduced for measurement of CSF t-tau, p-tau181, Aß42, and NfL. RESULTS: Twenty-seven iNPH, 27 AD, and 18 controls were included. The profiles of CSF t-tau, p-tau181, and t-tau/Aß42 in the iNPH and AD were significantly different (p < 0.0001). The profiles of CSF t-tau, p-tau181, and t-tau/Aß42 in the iNPH and control were not different (p > 0.05). Level of CSF Aß42 in iNPH was significantly lower than control (p < 0.0001) and also significantly higher than AD (p < 0.05). NfL level in iNPH and AD was increased, but its level in iNPH was significantly lower than that in AD (p = 0.005). NfL and t-tau level in the iNPH group was significantly correlated (coefficient = 0.649, p = 0.005), but not in AD (coefficient = 0.298, p = 0.157). CONCLUSION: Alzheimer's CSF biomarker profile of iNPH subjects showed moderately decreased Aß42 and normal t-tau, p-tau181, and t-tau/Aß42, which was distinguishable from AD. The different profiles and correlation of t-tau and NfL suggested different pathophysiology of AD and iNPH. t-tau was relatively an AD-specific neurodegenerative biomarker compared to NfL.

Genome-Wide Analysis Reveals Dynamic Epigenomic Differences in Soybean Response to Low-Phosphorus Stress.

Low-phosphorus (low-P) stress has a significant limiting effect on crop yield and quality. Although the molecular mechanisms of the transcriptional level responsible for the low-P stress response have been studied in detail, the underlying epigenetic mechanisms in gene regulation remain largely unknown. In this study, we evaluated the changes in DNA methylation, gene expression and small interfering RNAs (siRNAs) abundance genome-wide in response to low-P stress in two representative soybean genotypes with different P-efficiencies. The DNA methylation levels were slightly higher under low-P stress in both genotypes. Integrative methylation and transcription analysis suggested a complex regulatory relationship between DNA methylation and gene expression that may be associated with the type, region, and extent of methylation. Association analysis of low-P-induced differential methylation and gene expression showed that transcriptional alterations of a small part of genes were associated with methylation changes. Dynamic methylation alterations in transposable element (TE) regions in the CHH methylation context correspond with changes in the amount of siRNA under low-P conditions, indicating an important role of siRNAs in modulating TE activity by guiding CHH methylation in TE regions. Together, these results could help to elucidate the epigenetic regulation mechanisms governing the responses of plants to abiotic stresses.

Study on the In Vitro and In Vivo Antibacterial Activity and Biocompatibility of Novel TiN/Ag Multilayers Immobilized onto Biomedical Titanium.

To determine the short- and long-term antibacterial properties of a novel biomedical titanium alloy to ensure excellent biocompatibility of the TiN/Ag multilayers loaded with different doses of Ag+. First, nanosized TiN/Ag multilayers were accumulated onto titanium alloy (Ti-6Al-4V) substrates via multi-arc ion plating. Then, the multilayers were implanted with different doses of silver ions (1×1017 ions/cm², 1×1018 ions/cm², 5×1016 ions/cm², and 5×1017 ions/cm²). Both short- and long-term antibacterial properties against Streptococcus mutans and Staphylococcus aureus were assessed via unique methods. Additionally, the response and behaviors of MC3T3-E1 and L929 cells on the different surfaces were evaluated by a variety of methods through comparison to a normal matched substrate (Ti-6Al-4V). In Vitro and In Vivo analyses revealed that the multilayers containing different doses of Ag ions effectively prevented bacterial adhesion and eliminated the majority of adhered bacteria in the initial period. In addition, the antibacterial activity of each TiN/Ag group improved with time, with the antibacterial rate (Ra) ultimately reaching 99% (antibacterial activity: 1 × 1018 ions/cm² > 5 × 1017 ions/cm² > 1 × 1017 ions/cm² > 5 × 1016 ions/cm²). All of the samples loaded with Ag+ exhibited good compatibility, as well as higher cell proliferation and lower apoptosis than the pure Ti-6Al-4V substrates. Considering both bacteriostasis and biocompatibility, 1 × 1017 ions/cm² and 5 × 1017 ions/cm² are the recommended doses for orthopedic and dental implants. The results indicate that all of the samples loaded with Ag+ possess excellent biocompatibility and antibacterial activity against common bacteria that cause implantation infection. The samples loaded with Ag+ can be implanted into soft and hard growing tissues to greatly improve the survival rate of orthopedic and dental implants.