FBXO3 stabilizes USP4 and Twist1 to promote PI3K-mediated breast cancer metastasis.

Tumor metastasis is the major cause of breast cancer morbidity and mortality. It has been reported that the F-box protein FBXO3 functions as an E3 ubiquitin ligase in regulating various biological processes, including host autoimmune, antiviral innate immunity, and inflammatory response. However, the role of FBXO3 in tumor metastasis remains elusive. We have previously shown that ΔNp63α is a common inhibitory target in oncogene-induced cell motility and tumor metastasis. In this study, we show that FBXO3 plays a vital role in PI3K-mediated breast cancer metastasis independent of its E3 ligase activity and ΔNp63α in breast cancer cells and in mouse. FBXO3 can bind to and stabilize USP4, leading to Twist1 protein stabilization and increased breast cancer cell migration and tumor metastasis. Mechanistically, FBXO3 disrupts the interaction between USP4 and aspartyl aminopeptidase (DNPEP), thereby protecting USP4 from DNPEP-mediated degradation. Furthermore, p110αH1047R facilitates the phosphorylation and stabilization of FBXO3 in an ERK1-dependent manner. Knockdown of either FBXO3 or USP4 leads to significant inhibition of PI3K-induced breast cancer metastasis. Clinically, elevated expression of p110α/FBXO3/USP4/Twist1 is associated with poor overall survival (OS) and recurrence-free survival (RFS) of breast cancer patients. Taken together, this study reveals that the FBXO3-USP4-Twist1 axis is pivotal in PI3K-mediated breast tumor metastasis and that FBXO3/USP4 may be potential therapeutic targets for breast cancer treatment.

A chromosome-level genome assembly provides insights into Cornus wilsoniana evolution, oil biosynthesis, and floral bud development.

Cornus wilsoniana W. is a woody oil plant with high oil content and strong hypolipidemic effects, making it a valuable species for medicinal, landscaping, and ecological purposes in China. To advance genetic research on this species, we employed PacBio together with Hi-C data to create a draft genome assembly for C. wilsoniana. Based on an 11-chromosome anchored chromosome-level assembly, the estimated genome size was determined to be 843.51 Mb. The N50 contig size and N50 scaffold size were calculated to be 4.49 and 78.00 Mb, respectively. Furthermore, 30 474 protein-coding genes were annotated. Comparative genomics analysis revealed that C. wilsoniana diverged from its closest species ~12.46 million years ago (Mya). Furthermore, the divergence between Cornaceae and Nyssaceae occurred >62.22 Mya. We also found evidence of whole-genome duplication events and whole-genome triplication γ, occurring at ~44.90 and 115.86 Mya. We further inferred the origins of chromosomes, which sheds light on the complex evolutionary history of the karyotype of C. wilsoniana. Through transcriptional and metabolic analysis, we identified two FAD2 homologous genes that may play a crucial role in controlling the oleic to linoleic acid ratio. We further investigated the correlation between metabolites and genes and identified 33 MADS-TF homologous genes that may affect flower morphology in C. wilsoniana. Overall, this study lays the groundwork for future research aimed at identifying the genetic basis of crucial traits in C. wilsoniana.

Transcriptome Analysis Unveiled the Intricate Interplay between Sugar Metabolism and Lipid Biosynthesis in Symplocos paniculate Fruit.

Symplocos paniculate is an oil plant exhibiting tissue-specific variations in oil content and fatty acid composition across the whole fruit (mainly pulp and seed). And its oil synthesis is intricately linked to the accumulation and transformation of sugars. Nevertheless, there remains a dearth of understanding regarding how sugar metabolism impacts oil synthesis in S. paniculate fruit. To unravel the intricate mechanism underlying the impact of sugar metabolism on lipid biosynthesis in S. paniculata fruit, a comparative analysis was conducted on the transcriptome and metabolite content of pulp and seed throughout fruit development. The findings revealed that the impact of sugar metabolism on oil synthesis varied across different stages of fruit development. Notably, during the early fruit developmental stage (from 90 to 120 DAF), pivotal genes involved in sugar metabolism, such as PGK3, PKP1, PDH-E1, MDH, and malQ, along with key genes associated with oil synthesis like KAR, HAD, and PAP were predominantly expressed in the pulp. Consequently, this preferential expression led to earlier accumulation of oil in the pulp tissue compared to the seed. Whereas, during the fruit maturity stage (from 120 DAF to 140 DAF), these genes exhibited a high level of expression in seed, thereby facilitating the rapid and substantial accumulation of seed oil compared to pulp. The sugar metabolism activity in various parts of S. paniculata fruit plays a pivotal role in oil synthesis and is contingent upon the developmental stage. These findings can offer alternative genes for further gene enhancement through molecular biotechnology, thereby augmenting fruit oil yield and altering fatty acid composition.

Elucidation of Geniposide and Crocin Accumulation and Their Biosysnthsis-Related Key Enzymes during Gardenia jasminoides Fruit Growth.

Gardenia jasminoides fruits are extensively grown worldwide, with a large harvest, and its major medicinal ingredients are geniposide and crocins. Research on their accumulation and biosynthsis-related enzymes is rare. In this study, the accumulation of geniposide and crocin of G. jasminoides fruits at different developmental stages were clarified by HPLC. The highest cumulative amount of geniposide was 2.035% during the unripe-fruit period, and the highest content of crocin was 1.098% during the mature-fruit period. Furthermore, transcriptome sequencing was performed. A total of 50 unigenes encoding 4 key enzymes related in geniposide biosynthsis pathways were screened, and 41 unigenes encoding 7 key enzymes in the pathways of crocin were elucidated. It was found that the expression levels of differentially expressed genes of DN67890_c0_g1_i2-encoding GGPS, which is highly related to geniposide biosynthesis, and DN81253_c0_g1_i1-encoding lcyB, DN79477_c0_g1_i2-encoding lcyE, and DN84975_c1_g7_i11-encoding CCD, which are highly related to crocin biosynthesis, were consistent with the accumulation of geniposide and crocin content, respectively. The qRT-PCR results showed that the trends of relative expression were consistent with transcribed genes. This study provides insights for understanding the geniposide and crocin accumulation and biosynthsis during fruit development in G. jasminoides.

Differential expression analysis of sugar accumulation-related genes during chestnut nut development.

The chestnuts of Castanea mollissima Bl. are an important food crop in China, and have high nutritional content. To understand the pattern of sugar accumulation during chestnut nut development, the related enzyme gene regulatory pathways, and the molecular regulatory mechanisms of chestnut sugar biosynthesis metabolism, two chestnut varieties with different sugar content, namely Chengbu Youzhu (hereinafter referred to as CBYZ) and AnYou No. 1 (hereinafter referred to as AY01), were selected for investigation. Total sugar and starch content, and the activity of enzymes related to sugar accumulation, were measured in the nuts of the two chestnut varieties 10 days after flowering (DAF), 20 DAF, 30 DAF, 40 DAF, 50 DAF, 60 DAF, 70 DAF, 80 DAF, and 90 DAF. Changes in starch, straight-chain starch, and branched-chain starch content, and sucrose-phosphate synthase, soluble starch synthase, and granule-bound starch synthase enzyme activities were consistent with one-another. A total of 24 differentially expressed genes between the two varieties were associated with sugar biosynthesis and metabolism at three key stages (30, 60, and 90 DAF) of sugar accumulation. Further analysis showing upregulation of the expression of starch-related genes, such as ß-amylase, GYS, and INV indicated that these genes were not actively expressed in AY01, resulting in slow accumulation of starch and reduced sugar content. By contrast, the downregulation of the expression of genes, such as PGK and MDH1, indicated that these genes were actively expressed in low-sugar chestnuts, resulting in the rapid fermentation of sugars. A link between gene up- or down-regulation during different developmental stages of chestnut and the effect of their expression on sugar content were established by KEGG pathway enrichment analysis. These findings provide further insights into the mechanism of sugar biosynthesis in chestnuts.

Characterization and mechanism of seed dormancy in Symplocos paniculata.

Symplocos paniculata is a highly desirable oil species for biodiesel and premium edible oil feedstock. While germplasm preservation and breeding are crucial, the severity of seed dormancy poses a challenge to successful germination. We employed S. paniculata seeds as experimental materials and conducted an investigation into the types and causes of seed dormancy by analyzing the morphology and developmental characteristics of its embryo, exploring the water permeability property of the endocarp, and examining the presence of endogenous inhibitors, aiming to establish a theoretical foundation for overcoming seed dormancy and maximizing germplasm resource utilization. The findings revealed that the seed embryo had matured into a fully developed embryo, and no dormancy in terms of embryo morphology was observed. Upon reaching maturity, the endocarp of seeds undergoes significant lignification, resulting in notable differences in water absorption between cracked and intact seeds. The impermeability of the endocarp is one of the factors contributing to mechanical restriction. The different phases of endosperm extraction exerted varying effects on the germination of Chinese cabbage seeds, with the methanol phase exhibiting the most potent inhibitory effect. The presence of endogenous inhibitors emerged as the primary factor contributing to physiological dormancy in seeds. GC-MS analysis and validation trials revealed that fatty acids and phenolics, including hexadecanoic acid, oxadecanoic acid, and m-cresol, constituted the main types of endogenous inhibitory compounds found within the endosperm. These findings suggest that the seed dormancy in S. paniculata seeds has endocarp mechanical restriction, and the presence of endogenous inhibitors causes physiological dormancy.

Risk factors of elevated blood ammonia level in epilepsy patients treated with lamotrigine.

AbstractThe aim of this study was to explore the effect of lamotrigine (LTG) on blood ammonia level in patients with epilepsy and identify risk factors affecting blood ammonia level. This study included 91 epilepsy patients who were treated with LTG at Department of Neurology, Zhongshan Hospital, Xiamen University from January 2011 to April 2016, and were followed up for 3 years. Blood samples were taken during the interictal state and analyzed for blood LTG and ammonia levels. Total of 46.1% of the samples exceeded the median blood ammonia level, and 2.1% of patients had hyperammonemia. Blood ammonia level was positively correlated with LTG blood concentration. LTG combined with valproic acid therapy, seizure within 1 year, and elevated neutrophils affected blood ammonia level. Blood ammonia level was significantly correlated with plasma concentration of LTG. LTG combined with valproic acid therapy, seizure within 1 year, and elevated neutrophils may be risk factors for elevated blood ammonia level in epilepsy patients treated with LTG.

Comparison of Electroencephalography in Patients With Seizures Caused by Neurosyphilis and Viral Encephalitis.

Background: Neurosyphilis (NS) lacks specificity in clinical and imaging features, and patients are frequently misdiagnosed as viral encephalitis when they present with seizures. This study aimed to compare electroencephalography (EEG) in patients with seizures resulting from the two diseases and provide guidance for differential diagnosis. Methods: A retrospective study on patients diagnosed with neurosyphilis and viral encephalitis with seizures in the Department of Neurology, Zhongshan Hospital, Xiamen University from 2012 to 2020. Results: A total of 39 patients with seizures caused by neurosyphilis and 40 patients with seizures caused by viral encephalitis were included. Chi-square test analysis showed that compared with patients with viral encephalitis, patients with neurosyphilis mainly developed in middle-aged and elderly people (p < 0.001), were more likely to have temporal epileptiform discharges (p < 0.001), and less likely to have status epilepticus (SE) (p = 0.029). There was difference between two groups in the EEG performance of lateralized periodic discharges (LPDs) (p = 0.085). The two groups were matched for age and sex by case-control matching, and 25 cases in each group were successfully matched. Patients with neurosyphilis were more likely to have temporal epileptiform discharges than those with viral encephalitis (p = 0.002), and there were no significant differences in LPDs (p = 0.077) and SE (p = 0.088) between two groups. Conclusion: When EEG shows temporal epileptiform discharges, especially in the form of LPDs, we should consider the possibility of neurosyphilis.

Predictors of Outcome in Juvenile Myoclonic Epilepsy.

BACKGROUND: Juvenile myoclonic epilepsy (JME) is the most common idiopathic generalized epilepsy syndrome, accounting for 10% of all epilepsy. However, there is limited information regarding the predictors of seizure outcome. The aim of this study was to determine the predictors of seizure outcome in JME patients. METHODS: A population-based retrospective study of JME patients who were treated at the Department of Neurology of affiliated Zhongshan Hospital, Xiamen University from 2008 to 2013. RESULTS: Sixty-three patients (30 women and 33 men) were enrolled in this study. The median age at seizure onset was 14 years old, and the average duration of epilepsy was 5 years. The onset of JME at age <16 years was found in 63.5% of patients. The epileptiform runs ≥3 s were observed in 35.8% patients. Febrile seizure was noted in 28.9% of patients. Among 63 patients, 40 patients (63.5%) had remission. Multivariate analysis identified the following factors as significant predictors of seizure outcome: the onset of JME at age <16 years, epileptiform runs ≥3 s runs, and febrile seizure. CONCLUSION: The onset of JME at age <16 years, febrile seizures and epileptiform runs ≥3 s might be associated with poor long-term seizure outcome in patients with JME.