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Primary familial brain calcification (PFBC) is a genetic neurological disease, yet no effective treatment is currently available. Here, we identified five novel intronic variants in SLC20A2 gene from six PFBC families. Three of these variants increased aberrant SLC20A2 pre-mRNA splicing by altering the binding affinity of splicing machineries to newly characterized cryptic exons, ultimately causing premature termination of SLC20A2 translation. Inhibiting the cryptic-exon incorporation with splice-switching ASOs increased the expression levels of functional SLC20A2 in cells carrying SLC20A2 mutations. Moreover, by knocking in a humanized SLC20A2 intron 2 sequence carrying a PFBC-associated intronic variant, the SLC20A2-KI mice exhibited increased inorganic phosphate (Pi) levels in cerebrospinal fluid (CSF) and progressive brain calcification. Intracerebroventricular administration of ASOs to these SLC20A2-KI mice reduced CSF Pi levels and suppressed brain calcification. Together, our findings expand the genetic etiology of PFBC and demonstrate ASO-mediated splice modulation as a potential therapy for PFBC patients with SLC20A2 haploinsufficiency.
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MAIN CONCLUSION: We comprehensively identified and analyzed the Snf2 gene family. Some Snf2 genes were involved in responding to salt stress based on the RNA-seq and qRT-PCR analysis. Sucrose nonfermenting 2 (Snf2) proteins are core components of chromatin remodeling complexes that not only alter DNA accessibility using the energy of ATP hydrolysis, but also play a critical regulatory role in growth, development, and stress response in eukaryotes. However, the comparative study of Snf2 gene family in the six Brassica species in U's triangle model remains unclear. Here, a total of 405 Snf2 genes were identified, comprising 53, 50, and 46 in the diploid progenitors: Brassica rapa (AA, 2n = 20), Brassica nigra (BB, 2n = 16), and Brassica oleracea (CC, 2n = 18), and 93, 91, and 72 in the allotetraploid: Brassica juncea (AABB, 2n = 36), Brassica napus (AACC, 2n = 38), and Brassica carinata (BBCC, 2n = 34), respectively. These genes were classified into six clades and further divided into 18 subfamilies based on their conserved motifs and domains. Intriguingly, these genes showed highly conserved chromosomal distributions and gene structures, indicating that few dynamic changes occurred during the polyploidization. The duplication modes of the six Brassica species were diverse, and the expansion of most Snf2 in Brassica occurred primarily through dispersed duplication (DSD) events. Additionally, the majority of Snf2 genes were under purifying selection during polyploidization, and some Snf2 genes were associated with various abiotic stresses. Both RNA-seq and qRT-PCR analysis showed that the expression of BnaSnf2 genes was significantly induced under salt stress, implying their involvement in salt tolerance response in Brassica species. The results provide a comprehensive understanding of the Snf2 genes in U's triangle model species, which will facilitate further functional analysis of the Snf2 genes in Brassica plants.
Subject(s)
Brassica , Gene Expression Regulation, Plant , Plant Proteins , Salt Stress , Brassica/genetics , Brassica/physiology , Plant Proteins/genetics , Plant Proteins/metabolism , Salt Stress/genetics , Multigene Family , Phylogeny , Genome, Plant/genetics , Gene Expression ProfilingABSTRACT
Acyl carrier proteins (ACPs) have been reported to play a crucial role in responding to biotic and abiotic stresses, regulating growth and development. However, the biological function of the ACP gene family in the Brassica genus has been limited until now. In this study, we conducted a comprehensive analysis and identified a total of 120 ACP genes across six species in the Brassica genus. Among these, there were 27, 26, and 30 ACP genes in the allotetraploid B. napus, B. juncea, and B. carinata, respectively, and 14, 13, and 10 ACP genes in the diploid B. rapa, B. oleracea, and B. nigra, respectively. These ACP genes were further classified into six subclades, each containing conserved motifs and domains. Interestingly, the majority of ACP genes exhibited high conservation among the six species, suggesting that the genome evolution and polyploidization processes had relatively minor effects on the ACP gene family. The duplication modes of the six Brassica species were diverse, and the expansion of most ACPs in Brassica occurred primarily through dispersed duplication (DSD) events. Furthermore, most of the ACP genes were under purifying selection during the process of evolution. Subcellular localization experiments demonstrated that ACP genes in Brassica species are localized in chloroplasts and mitochondria. Cis-acting element analysis revealed that most of the ACP genes were associated with various abiotic stresses. Additionally, RNA-seq data revealed differential expression levels of BnaACP genes across various tissues in B. napus, with particularly high expression in seeds and buds. qRT-PCR analysis further indicated that BnaACP genes play a significant role in salt stress tolerance. These findings provide a comprehensive understanding of ACP genes in Brassica plants and will facilitate further functional analysis of these genes.
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BACKGROUND: Calcium-dependent protein kinases (CPKs) are crucial for recognizing and transmitting Ca2+ signals in plant cells, playing a vital role in growth, development, and stress response. This study aimed to identify and detect the potential roles of the CPK gene family in the amphidiploid Brassica carinata (BBCC, 2n = 34) using bioinformatics methods. RESULTS: Based on the published genomic information of B. carinata, a total of 123 CPK genes were identified, comprising 70 CPK genes on the B subgenome and 53 on the C subgenome. To further investigate the homologous evolutionary relationship between B. carinata and other plants, the phylogenetic tree was constructed using CPKs in B. carinata and Arabidopsis thaliana. The phylogenetic analysis classified 123 family members into four subfamilies, where gene members within the same subfamily exhibited similar conserved motifs. Each BcaCPK member possesses a core protein kinase domain and four EF-hand domains. Most of the BcaCPK genes contain 5 to 8 introns, and these 123 BcaCPK genes are unevenly distributed across 17 chromosomes. Among these BcaCPK genes, 120 replicated gene pairs were found, whereas only 8 genes were tandem duplication, suggesting that dispersed duplication mainly drove the family amplification. The results of the Ka/Ks analysis indicated that the CPK gene family of B. carinata was primarily underwent purification selection in evolutionary selection. The promoter region of most BcaCPK genes contained various stress-related cis-acting elements. qRT-PCR analysis of 12 selected CPK genes conducted under cadmium and salt stress at various points revealed distinct expression patterns among different family members in response to different stresses. Specifically, the expression levels of BcaCPK2.B01a, BcaCPK16.B02b, and BcaCPK26.B02 were down-regulated under both stresses, whereas the expression levels of other members were significantly up-regulated under at least one stress. CONCLUSION: This study systematically identified the BcaCPK gene family in B. carinata, which contributes to a better understanding the CPK genes in this species. The findings also serve as a reference for analyzing stress responses, particularly in relation to cadmium and salt stress in B. carinata.
Subject(s)
Brassica , Brassica/genetics , Phylogeny , Cadmium/metabolism , Multigene Family , Genomics , Gene Expression Regulation, Plant , Stress, Physiological/genetics , Plant Proteins/genetics , Genome, PlantABSTRACT
Objectives: Serous microcystic adenoma (SMA), a primary benign pancreatic tumor which can be clinically followed-up instead of undergoing surgery, are sometimes mis-distinguished as pancreatic neuroendocrine tumor (pNET) in regular preoperative imaging examinations. This study aimed to analyze preoperative contrast-enhanced ultrasound (CEUS) and shear wave elastography (SWE) features of SMAs in comparison to pNETs. Material and methods: In this retrospective study, patients with imaging-diagnosed pancreatic lesions were screened between October 2020 to October 2022 (ethical approval No. B2020-309R). Performing by a Siemens Sequoia (Siemens Medical Solutions, Mountain View, CA, USA) equipped with a 5C-1 curved array transducer (3.0-4.5 MHz), CEUS examination was conducted to observe the microvascular perfusion patterns of pancreatic lesions in arterial phase, venous/late phases (VLP) using SonoVue® (Bracco Imaging Spa, Milan, Italy) as the contrast agent. Virtual touch tissue imaging and quantification (VTIQ) - SWE was used to measure the shear wave velocity (SWV, m/s) value to represent the quantitative stiffness of pancreatic lesions. Multivariate logistic regression was performed to analyze potential ultrasound and clinical features in discriminating SMAs and pNETs. Results: Finally, 30 SMA and 40 pNET patients were included. All pancreatic lesions were pathologically proven via biopsy or surgery. During the arterial phase of CEUS, most SMAs and pNETs showed iso- or hyperenhancement (29/30, 97 % and 31/40, 78 %), with a specific early honeycomb enhancement pattern appeared in 14/30 (47 %) SMA lesions. During the VLP, while most of the SMA lesions remained iso- or hyperenhancement (25/30, 83 %), nearly half of the pNET lesions revealed an attenuated hypoenhancement (17/40, 43 %). The proportion of hypoenhancement pattern during the VLP of CEUS differed significantly between SMAs and pNETs (P = 0.021). The measured SWV value of SMAs was significantly higher than pNETs (2.04 ± 0.70 m/s versus 1.42 ± 0.44 m/s, P = 0.002). Taking a SWV value > 1.83 m/s as a cutoff in differentiating SMAs and pNETs, the area under the receiver operating characteristic curve (AUROC) was 0.825, with sensitivity, specificity and likelihood ratio (+) of 85.71 %, 72.73 % and 3.143, respectively. Multivariate logistic regression revealed that SWV value (m/s) of the pancreatic lesion was an independent variable in discriminating SMA and pNET. Conclusion: By comprehensively evaluating CEUS patterns and SWE features, SMA and pNET may be well differentiated before the operation. While SMA typically presents as harder lesion in VTIQ-SWE, exhibiting a specific honeycomb hyperenhancement pattern during the arterial phase of CEUS, pNET is characterized by relative softness, occasionally displaying a wash-out pattern during the VLP of CEUS.
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Cadmium (Cd), which is a nonessential heavy metal element for organisms, can have a severe impact on the growth and development of organisms that absorb excessive Cd. Studies have shown that Brassica carinata, a semiwild oil crop, has strong tolerance to various abiotic stresses, and RNA-seq has revealed that the B. carinata superoxide dismutase gene (BcaSOD1) likely responds to Cd stress. To elucidate the BcaSOD1 function involved in tolerance of Cd stress, we cloned the coding sequences of BcaSOD1 from a purple B. carinata accession and successfully transferred it into Arabidopsis thaliana. The subcellular localization results demonstrated that BcaSOD1 was primarily located in the plasma membrane, mitochondria and nucleus. Overexpression of BcaSOD1 in transgenic Arabidopsis (OE) effectively decreased the toxicity caused by Cd stress. Compared to the WT (wild type lines), the OE lines exhibited significantly increased activities of antioxidant enzymes (APX, CAT, POD, and SOD) after exposure to 2.5 mM CdCl2. The Cd content of underground (root) in the OE line was dominantly higher than that in the WT; however, the Cd content of aboveground (shoot) was comparable between the OE and WT types. Moreover, the qRTâPCR results showed that several heavy metal detoxification-related genes (AtIREG2, AtMTP3, AtHMA3, and AtNAS4) were significantly upregulated in the roots of OE lines under Cd treatment, suggesting that these genes are likely involved in Cd absorption in the roots of OE lines. In addition, both comparable transcriptome and qRT-PCR analyses revealed that exogenous BcaSOD1 noticeably facilitates detoxification by stimulating the expression of two arginine (Arg) biosynthesis genes (AtGDH1 and AtGDH2) while inhibiting the expression of AtARGAH1, a negative regulator in biosynthesis of Arg. The Arg content was subsequently confirmed to be significantly enhanced in OE lines under Cd treatment, indicating that BcaSOD1 likely strengthened Cd tolerance by regulating the expression of Arg-related genes. This study demonstrates that BcaSOD1 can enhance Cd tolerance and reveals the molecular mechanism of this gene, providing valuable insights into the molecular mechanism of Cd tolerance in plants.
Subject(s)
Arabidopsis , Metals, Heavy , Cadmium/toxicity , Cadmium/metabolism , Arabidopsis/metabolism , Superoxide Dismutase/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Metals, Heavy/toxicity , Metals, Heavy/metabolism , Arginine/genetics , Arginine/metabolismABSTRACT
Soil salinization, an intractable problem, is becoming increasingly serious and threatening fragile natural ecosystems and even the security of human food supplies. Sorghum (Sorghum bicolor L.) is one of the main crops growing in salinized soil. However, the tolerance mechanisms of sorghum to saline-alkaline soil are still ambiguous. In this study, RNA sequencing was carried out to explore the gene expression profiles of sorghum treated with sodium bicarbonate (150 mM, pH = 8.0, treated for 0, 6, 12 and 24 h). The results show that 6045, 5122, 6804, 7978, 8080 and 12,899 differentially expressed genes (DEGs) were detected in shoots and roots after 6, 12 and 24 h treatments, respectively. GO, KEGG and weighted gene co-expression analyses indicate that the DEGs generated by saline-alkaline stress were primarily enriched in plant hormone signal transduction, the MAPK signaling pathway, starch and sucrose metabolism, glutathione metabolism and phenylpropanoid biosynthesis. Key pathway and hub genes (TPP1, WRKY61, YSL1 and NHX7) are mainly related to intracellular ion transport and lignin synthesis. The molecular and physiological regulation processes of saline-alkali-tolerant sorghum are shown by these results, which also provide useful knowledge for improving sorghum yield and quality under saline-alkaline conditions.
Subject(s)
Sorghum , Transcriptome , Humans , Sorghum/genetics , Ecosystem , Gene Expression Profiling , Soil/chemistry , Gene Expression Regulation, Plant , Stress, Physiological/geneticsABSTRACT
Introduction: Research has demonstrated that cognitive flexibility is associated with academic achievement, with poorer cognitive flexibility being linked to poorer academic performance. Strategy conversion is an example of cognitive flexibility, which requires individuals to quickly and flexibly switch between strategies depending on the task at hand. Studies have investigated the impact of emotional motivation on cognitive flexibility, with varying results. Furthermore, research has indicated that a high task load increases psychological burden and reduces cognitive flexibility, but few studies have analyzed the impact of task load on the relationship between emotional motivation and cognitive flexibility. This study sought to investigate the effect of emotional motivation on cognitive flexibility based on strategy switching, and the moderating effect of task load. Methods: Three experiments were conducted. Experiment 1 used forced conversion paradigm with a moderate task load that required participants to estimate tasks using a given strategy. Experiment 2 employed matched conversion paradigm with a high task load, informing participants of the strategies to be used but without any clues, necessitating the selection and execution of appropriate strategies based on the question features. Experiment 3 adopted free conversion paradigm with a relatively low task load, allowing participants to freely choose and execute strategies without any correctness or error criteria. Results: The intensity and direction of emotional motivation have a complicated and fluctuating impact on the flexibility to utilize strategies. When the task workload is high, the intensity of emotional motivation has a significant impact on strategy utilization flexibility, with low approach motivation being more conducive to flexibility. Conversely, when the task workload is low, the direction of emotional motivation has a greater effect, with high avoidance motivation being more advantageous. Discussion: This study demonstrated that high workload could bring about a low intensity advantage, while low workload could induce an avoidance direction advantage, suggesting that task load could moderate the impact of emotional motivation on arithmetic strategy utilization flexibility, and avoidance motivation is not always detrimental to cognitive flexibility.
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Objective: Excessive obesity can lead to dysfunction in adipose tissue, which contributes to the development of comorbidities associated with obesity, such as type 2 diabetes (T2D), cardiovascular and cerebrovascular disease, among others. Previous research has mainly focused on the Vanin family in systemic inflammatory diseases or predicting its role in tumor prognosis, while neglecting its role as a secretory protein in adipose tissue inflammation and metabolism. The objective of this study was to compare the changes in Vanin-2 levels in the circulating blood of normal and obese individuals, and to assess its correlation with inflammatory factors in vivo. Furthermore, the study aimed to systematically evaluate its effectiveness in human weight loss surgery. Methods: Serum concentrations of Vanin-2 and inflammatory indicators were measured in 518 volunteers. Furthermore, the concentrations of Vanin-2 were measured both before and after weight loss through a dietetic program or laparoscopic sleeve gastrectomy (LSG). Additionally, we assessed the levels of insulin, adiponectin, and inflammation-related factors. The hormonal profile and changes in body weight were evaluated at baseline and 3 months after surgery. Results: Serum levels of Vanin-2 were found to be significantly increased in individuals with overweight/obesity (OW/OB) group (controls 438.98 ± 72.44, OW/OB 530.89 ± 79.39 ug/L; p < 0.001). These increased levels were associated with IL-18, BMI, FAT%, and HOMA-IR. However, levels of Vanin-2 remained unchanged after conventional dietary treatment. On the other hand, weight loss induced by LSG resulted in a significant decrease in Vanin-2 concentrations from 586.44 ± 48.84 to 477.67 ± 30.27 ug/L (p < 0.001), and this decrease was associated with the Vanin-2 concentrations observed before the operation. Conclusion: Serum Vanin-2 is a highly effective biomarker for assessing adipose tissue inflammation in obesity and has the potential to serve as a predictor of bariatric surgery outcomes.
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Arabis paniculata is a newly discovered hyperaccumulator known for its ability to accumulate multiple metals. WRKY proteins play a significant role in plant responses to various stresses, including cadmium (Cd) stress. However, there is limited research on the molecular biology of Arabis paniculata, especially regarding the WRKY family. In this study, we conducted third-generation sequencing for functional annotation and structural analysis of Arabis paniculata. We obtained 41,196 high-quality isoforms from the full-length transcriptome, with an average length of 1043 bp. A total of 26,670 genes were predicted against NR, Swissprot, KOG, and KEGG databases. Functional comparison using the KOG database revealed excellent annotation in 25 functional categories, with general function prediction (1822 items) being the most predominant. MISA analysis identified 12,593 SSR loci, with single nucleotide repeats being the largest category (44.83% of the total). Moreover, our predictions provide insights into 20,022 coding sequences (CDS), 811 transcription factors, and 17,963 LncRNAs. In total, 34 WRKY gene sequences were identified in Arabis paniculata. Bioinformatics analysis revealed diverse numbers of amino acids in these WRKYs (113 to 545 aa), and a conserved WRKYGQK sequence within the N-terminus of the WRKY protein. Furthermore, all WRKYs were found to be localized in the nucleus. Phylogenetic analysis classified the WRKY genes into three categories: I (14 members), II (17 members), and III (3 members). Category II was subsequently divided into four sub-categories: II-a (8 members), II-b (1 member), II-c (1 member), and II-d (7 members). Our quantitative real-time polymerase chain reaction (qRT-PCR) experiments revealed that ApWRKY23 and ApWRKY34 exhibited the highest expression levels at the 24-h time point, suggesting their potential role as the candidate genes for Cd stress response. These findings contribute to our understanding of the genomic information of Arabis paniculata and provide a basis for the analysis of its genetic diversity. Additionally, this study paves the way for a comprehensive exploration of the molecular mechanisms underlying the WRKY genes in Arabis paniculata under Cd stress conditions.
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Cadmium (Cd) is a highly toxic heavy metal with severe impacts on plant growth and development. Although a multitude of plants have acquired strong tolerance to Cd stress, the underlying molecular mechanism has not been fully elucidated. Here, we identified a Agamous-like MADS-box gene (EcAGL) from Erigeron canadensis. The expression of EcAGL was obviously raised under Cd stress and subcellular localization indicated EcAGL was localized in the nucleus. Overexpression of EcAGL in Arabidopsis thaliana showed marked alleviation of the Cd-induced reduction; Compared to wild-type lines, the antioxidant enzymes activities were increased in EcAGL overexpressing lines under Cd stress. The roots Cd content of transgenic lines was not different with the control plants, whereas significant reduction in shoots Cd content was detected in the transgenic lines, indicating that this gene can enhance Cd tolerance by reducing Cd accumulation in Arabidopsis. Moreover, the expression levels of heavy metal ATPase (AtHMA2 and AtHMA3) and natural resistance-associated macrophage protein (AtNRAMP5) genes in the root of transgenic lines decreased under Cd stress, indicating that EcAGL likely hampered the Cd transport pathway. Gene expression profiles in shoot showed that EcAGL likely modulates the expression of 1-aminocyclopropane-1-carboxylic acid synthase gene (AtACS2), which is involved in the ethylene synthesis pathway, to strengthen the tolerance to Cd. Collectively, these results indicate that EcAGL plays a significant role in regulating Cd tolerance in E. canadensis by alleviating oxidative stress, Cd transport and affecting the ethylene biosynthesis pathway, providing new insight into the molecular mechanism underlying plant tolerance to Cd stress.
Subject(s)
Arabidopsis Proteins , Arabidopsis , Metals, Heavy , Arabidopsis/metabolism , Cadmium/toxicity , Cadmium/metabolism , Plants, Genetically Modified/genetics , Arabidopsis Proteins/metabolism , Metals, Heavy/metabolism , Ethylenes/metabolism , Antioxidants/metabolism , Plant Roots/metabolism , Gene Expression Regulation, PlantABSTRACT
Aim: To investigate the diagnostic value of resistance index (RI) in differentiating focal liver lesions. Patients and methods: In this retrospective study, a total of 576 patients with histologically confirmed focal liver lesions were included. Each patient underwent B-mode ultrasound examination and color Doppler ultrasound examination. The RI values of different focal liver lesions were recorded and compared. Results: The mean RI value of benign lesions was significantly lower than that of malignant lesions (0.54 ± 0.10 vs. 0.71 ± 0.12) (p <0.05). In malignant lesions, the RI value of intrahepatic cholangiocarcinoma was significantly lower than that of hepatocellular carcinoma lesions. Furthermore, in hepatocellular carcinoma lesions, the RI of large lesions (group 4: >10 cm) was significantly lower than that of small lesions (group 1: ≤2 cm, group 2: 2-5 cm) (p <0.05). Taken RI of 0.615 as a cutoff value to differentiate malignant and benign lesions, the sensitivity, specificity, positive predictive value and negative predictive value were 82.80%, 81.00%, 81.34% and 82.48%, respectively. Conclusion: Color Doppler ultrasound examination is a valuable imaging method in detecting blood flow signal within liver lesions. The RI parameter should be helpful in differentiating malignant and benign liver tumors.
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OBJECTIVES: The aim of this study was to modify recognized clinically relevant post-operative pancreatic fistula (CR-POPF) risk evaluation models with quantitative ultrasound shear wave elastography (SWE) values and identified clinical parameters to improve the objectivity and reliability of the prediction. METHODS: Two prospective, successive cohorts were initially designed for the establishment of CR-POPF risk evaluation model and the internal validation. Patients who scheduled to receive pancreatectomy were enrolled. Virtual touch tissue imaging and quantification (VTIQ)-SWE was used to quantify pancreatic stiffness. CR-POPF was diagnosed according to 2016 International Study Group of Pancreatic Fistula standard. Recognized peri-operative risk factors of CR-POPF were analyzed, and the independent variables selected from multivariate logistic regression were used to build the prediction model. RESULTS: Finally, the CR-POPF risk evaluation model was built in a group of 143 patients (cohort 1). CR-POPF occurred in 52/143 (36%) patients. Constructed from SWE values and other identified clinical parameters, the model achieved an area under the receiver operating characteristic curve of 0.866, with sensitivity, specificity, and likelihood ratio of 71.2%, 80.2%, and 3.597 in predicting CR-POPF. Decision curve of modified model revealed a better clinical benefit compared to the previous clinical prediction models. The models were then examined via internal validation in a separate collection of 72 patients (cohort 2). CONCLUSIONS: Risk evaluation model based on SWE and clinical parameters is a potential non-invasive way to pre-operatively, objectively predict CR-POPF after pancreatectomy. CLINICAL RELEVANCE STATEMENT: Our modified model based on ultrasound shear wave elastography may provide an easy access in pre-operative and quantitative evaluating the risk of CR-POPF following pancreatectomy and improve the objectivity and reliability of the prediction compared to previous clinical models. KEY POINTS: ⢠Modified prediction model based on ultrasound shear wave elastography (SWE) provides an easy access for clinicians to pre-operatively, objectively evaluate the risk of clinically relevant post-operative pancreatic fistula (CR-POPF) following pancreatectomy. ⢠Prospective study with validation showed that the modified model provides better diagnostic efficacy and clinical benefits compared to previous clinical models in predicting CR-POPF. ⢠Peri-operative management of CR-POPF high-risk patients becomes more possible.
Subject(s)
Elasticity Imaging Techniques , Pancreatectomy , Humans , Pancreatectomy/adverse effects , Pancreatectomy/methods , Pancreatic Fistula/epidemiology , Pancreatic Fistula/etiology , Prospective Studies , Elasticity Imaging Techniques/methods , Reproducibility of Results , Risk Factors , Postoperative Complications/diagnostic imaging , Postoperative Complications/etiology , Pancreaticoduodenectomy/adverse effects , Retrospective StudiesABSTRACT
Soil and water are increasingly at risk of contamination from the toxic heavy metals lead (Pb) and cadmium (Cd). Arabis paniculata (Brassicaceae) is a hyperaccumulator of heavy metals (HMs) found widely distributed in areas impacts by mining activities. However, the mechanism by which A. paniculata tolerates HMs is still uncharacterized. For this experiment, we employed RNA sequencing (RNA-seq) in order to find Cd (0.25 mM)- and Pb (2.50 mM)-coresponsive genes A. paniculata. In total, 4490 and 1804 differentially expressed genes (DEGs) were identified in root tissue, and 955 and 2209 DEGs were identified in shoot tissue, after Cd and Pb exposure, respectively. Interestingly in root tissue, gene expression corresponded similarly to both Cd and Pd exposure, of which 27.48% were co-upregulated and 41.00% were co-downregulated. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses showed that the co-regulated genes were predominantly involved in transcription factors (TFs), cell wall biosynthesis, metal transport, plant hormone signal transduction, and antioxidant enzyme activity. Many critical Pb/Cd-induced DEGs involved in phytohormone biosynthesis and signal transduction, HM transport, and transcription factors were also identified. Especially the gene ABCC9 was co-downregulated in root tissues but co-upregulated in shoot tissues. The co-downregulation of ABCC9 in the roots prevented Cd and Pb from entering the vacuole rather than the cytoplasm for transporting HMs to shoots. While in shoots, the ABCC9 co-upregulated results in vacuolar Cd and Pb accumulation, which may explain why A. paniculata is a hyperaccumulator. These results will help to reveal the molecular and physiological processes underlying tolerance to HM exposure in the hyperaccumulator A. paniculata, and aid in future efforts to utilize this plant in phytoremediation.
Subject(s)
Arabis , Metals, Heavy , Soil Pollutants , Cadmium/metabolism , Arabis/genetics , Arabis/metabolism , Lead/analysis , Transcriptome , Metals, Heavy/analysis , Biodegradation, Environmental , Plant Growth Regulators/metabolism , Soil Pollutants/analysis , Plant Roots/metabolismABSTRACT
OBJECTIVE: This study seeks to evaluate the value of the high-frame-rate vector flow imaging technique in assessing the hemodynamic changes of carotid atherosclerotic stenosis in aging people (>60 years old). METHODS: Aging patients diagnosed with carotid atherosclerotic stenosis who underwent carotid high-frame-rate vector flow imaging examination were prospectively enrolled. A Mindray Resona7s ultrasound machine equipped with high-frame-rate vector flow function was used for ultrasound evaluation. First, B mode ultrasound and color Doppler flow imaging were used to evaluate carotid stenosis. Then, the vector arrows and flow streamline detected by V Flow were analyzed and the wall shear stress values (Pa) at the carotid stenosis site were measured. All patients were divided into symptomatic and asymptomatic groups according to whether they had acute/subacute stroke or other clinical symptoms within 2 weeks before ultrasound examination. The results of digital subtraction angiography or computed tomography angiography were used as the gold standard. The stenosis rate was calcified, according to North American Symptomatic Carotid Endarterectomy Trial criteria. The diagnostic values of wall shear stress, conventional ultrasound, and the combined diagnosis in carotid atherosclerotic stenosis were compared. RESULTS: Finally, 88 patients with carotid atherosclerotic plaque were enrolled (71 males (80.7%), mean age 67.6 ± 5.4 years). The success rate of high-frame-rate vector flow imaging was 96.7% (88/91). The WSS value of symptomatic carotid stenosis (1.4 ± 0.15 Pa) was significantly higher than that of asymptomatic carotid stenosis (0.80 ± 0.08 Pa) (p < 0.05). Taking the wall shear stress value > 0.78 Pa as the diagnostic criteria for symptomatic carotid atherosclerotic plaque, the area under receiver operating characteristic curves was 0.79 with 87.1% sensitivity and 69.6% specificity. The area under receiver operating characteristic curves of the combined diagnosis (0.966) for differentiating severe carotid atherosclerotic stenosis was significantly higher than that of conventional ultrasound and WSS value, with 89.7% sensitivity and 93.2% specificity (p < 0.05). CONCLUSION: As a non-invasive imaging method, the high-frame-rate vector flow imaging technique showed potential value in the preoperative assessment of the symptomatic carotid atherosclerotic stenosis and diagnosing carotid atherosclerotic stenosis in aging patients.
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OBJECTIVE: To explore the value of vascular architecture detected by micro vascular imaging (MVI) in preoperative diagnosis of focal liver lesions (FLLs). METHODS: In this retrospective study, patients with surgery and histopathologically proved or radiologically confirmed FLLs were included. Vascular architecture of FLLs were acquired by color Doppler flow imaging (CDFI) and MVI on LOGIQ™ E20 ultrasound machine (C1-6 convex array probes). Alder semiquantitative analysis (grade 0-3) and morphologic features of blood vessels (pattern a-f) were used to assess the blood flow within the FLLs. Interobserver agreement for evaluating blood flow of FLLs was analyzed. Using Adler's grading or morphologic patterns as diagnostic criteria for malignant FLLs, the diagnostic efficiency was analyzed and compared. RESULTS: From October 2021 and February 2022, 50 patients diagnosed with 40 malignant FLLs and 10 benign FLLs were finally included. The Kappa value within two observers for evaluating the blood flow of FLLs was 0.78 for MVI and 0.55 for CDFI. According to Alder semiquantitative analysis, more high-level blood flow signals (grade 2-3) were detected by MVI than CDFI (Pâ<â0.05). Based on high-level blood flow signals (grade 2-3) and hypervascular supply patterns (pattern e and f), the diagnostic accuracy for malignant FLLs were 76% and 68% for MVI, 56% and 38% for CDFI, respectively. CONCLUSION: MVI is superior to CDFI in evaluating vascular architecture of FLLs. The high-level flow signals and hypervascular pattern detected by MVI have a useful and complementary value in preoperative non-invasive identification of malignant FLLs.
Subject(s)
Liver Neoplasms , Humans , Liver Neoplasms/blood supply , Retrospective Studies , Liver/diagnostic imaging , Liver/pathology , Ultrasonography , Microvessels/diagnostic imaging , Sensitivity and Specificity , Diagnosis, DifferentialABSTRACT
OBJECTIVES: To explore the potential added value of dynamic contrast enhanced ultrasound (DCE-US) using VueBox® software for the diagnosis of small solid thyroid nodules (≤1.0âcm). PATIENTS AND METHODS: This prospective study was approved by the institutional review board and it was performed at two hospitals from January 2020 to October 2020. B mode ultrasound and contrast enhanced ultrasound (CEUS) images were obtained for 79 small solid thyroid nodules (≤1.0âcm) confirmed by ultrasound-guided fine needle aspiration cytology results in 79 consecutive patients (55 women and 24 men, median age: 41 years). The CEUS time-intensity curves (TICs) of thyroid nodules and surrounding parenchyma were created by VueBox® software (Bracco, Italy). The CEUS quantitative parameters were obtained after curve fitting. The diagnostic performance of CEUS and DCE-US was evaluated and compared. The weighted kappa statistic (κ) was performed to assess the interobserver agreement and consistency between the diagnosis of CEUS and DCE-US. RESULTS: Among the 79 thyroid nodules, 56 (70.9 %) were malignant and 23 (29.1 %) were benign lesions. Hypoenhancement during the arterial phase of CEUS was associated with malignancy (Pâ<â0.001), with an AUC of 0.705 (sensitivity 71.4 %, specificity 69.6 %). Among all CEUS quantitative parameters, the peak enhancement (PE), wash-in rate (WiR), and wash-out rate (WoR) of DCE-US in malignancies were significantly lower than those in benign nodules (Pâ=â0.049, Pâ=â0.046, and Pâ=â0.020, respectively). The area under the receiver operating characteristic curves (AUCs) of PE, WiR, and WoR were 0.642 (sensitivity 65.2 %, specificity 67.9 %), 0.643 (sensitivity 43.5 %, specificity 91.1 %), and 0.667 (sensitivity 69.6 %, specificity 69.6 %) in differentiation between benign and malignant small solid thyroid nodules (≤1.0âcm), respectively. Comparing the quantitative parameters of DCE-US between small solid thyroid nodules and surrounding normal thyroid parenchyma, the PE, WiAUC, WiR, wash-in perfusion index (WiPI), WoAUC, WiWoAUC, and WoR of the nodules were significantly lower than those of normal thyroid tissue (Pâ=â0.008, Pâ<â0.001, Pâ=â0.037, Pâ=â0.009, Pâ=â0.003, Pâ=â0.002, Pâ=â0.049, respectively). A total of 16 (20.3 %) nodules showed isoenhancement during the arterial phase of CEUS, while the median PE ratio of surrounding tissue and thyroid nodules was 1.70 (IQR: 1.33-1.89). CONCLUSIONS: VueBox® is a helpful tool for the evaluation of dynamic microvascularization of thyroid nodules, and DCE-US using VueBox® perfusion analysis could provide added values for differential diagnosis of small solid thyroid nodules (≤1.0âcm).
Subject(s)
Thyroid Nodule , Male , Humans , Female , Adult , Thyroid Nodule/diagnostic imaging , Thyroid Nodule/pathology , Prospective Studies , Contrast Media , Perfusion/methods , Ultrasonography/methods , Diagnosis, DifferentialABSTRACT
Brain calcification is a critical aging-associated pathology and can cause multifaceted neurological symptoms. Cerebral phosphate homeostasis dysregulation, blood-brain barrier defects, and immune dysregulation have been implicated as major pathological processes in familial brain calcification (FBC). Here, we analyzed two brain calcification families and identified calcification co-segregated biallelic variants in the CMPK2 gene that disrupt mitochondrial functions. Transcriptome analysis of peripheral blood mononuclear cells (PBMCs) isolated from these patients showed impaired mitochondria-associated metabolism pathways. In situ hybridization and single-cell RNA sequencing revealed robust Cmpk2 expression in neurons and vascular endothelial cells (vECs), two cell types with high energy expenditure in the brain. The neurons in Cmpk2-knockout (KO) mice have fewer mitochondrial DNA copies, down-regulated mitochondrial proteins, reduced ATP production, and elevated intracellular inorganic phosphate (Pi) level, recapitulating the mitochondrial dysfunction observed in the PBMCs isolated from the FBC patients. Morphologically, the cristae architecture of the Cmpk2-KO murine neurons was also impaired. Notably, calcification developed in a progressive manner in the homozygous Cmpk2-KO mice thalamus region as well as in the Cmpk2-knock-in mice bearing the patient mutation, thus phenocopying the calcification pathology observed in the patients. Together, our study identifies biallelic variants of CMPK2 as novel genetic factors for FBC; and demonstrates how CMPK2 deficiency alters mitochondrial structures and functions, thereby highlighting the mitochondria dysregulation as a critical pathogenic mechanism underlying brain calcification.
ABSTRACT
Cadmium (Cd) is a highly toxic pollutant in soil and water that severely hampers the growth and reproduction of plants. Phytoremediation has been presented as a cost-effective and eco-friendly method for addressing heavy metal pollution. However, phytoremediation is restricted by the limited number of accumulators and the unknown mechanisms underlying heavy metal tolerance. In this study, we demonstrated that Erigeron canadensis (Asteraceae), with its strong adaptability, is tolerant to intense Cd stress (2 mmol/L CdCl2 solution). Moreover, E. canadensis exhibited a strong ability to accumulate Cd2+ when treated with CdCl2 solution. The activity of some antioxidant enzymes, as well as the malondialdehyde (MDA) level, was significantly increased when E. canadensis was treated with different CdCl2 solutions (0.5, 1, 2 mmol/L CdCl2). We found high levels of superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities under 1 mmol/L CdCl2 treatment. Comparative transcriptomic analysis identified 5,284 differentially expressed genes (DEGs) in the roots and 3,815 DEGs in the shoots after E. canadensis plants were exposed to 0.5 mM Cd. Functional annotation of key DEGs indicated that signal transduction, hormone response, and reactive oxygen species (ROS) metabolism responded significantly to Cd. In particular, the DEGs involved in auxin (IAA) and ethylene (ETH) signal transduction were overrepresented in shoots, indicating that these genes are mainly involved in regulating plant growth and thus likely responsible for the Cd tolerance. Overall, these results not only determined that E. canadensis can be used as a potential accumulator of Cd but also provided some clues regarding the mechanisms underlying heavy metal tolerance.