Genome-wide identification and functional characterization of the PheE2F/DP gene family in Moso bamboo.

BACKGROUND: E2F/DP proteins have been shown to regulate genes implicated in cell cycle control and DNA repair. However, to date, research into the potential role of the Moso bamboo E2F/DP family has been limited. RESULTS: Here, we identified 23 E2F/DPs in the Moso bamboo genome, including nine E2F genes, six DP genes, eight DEL genes and one gene with a partial E2F domain. An estimation of the divergence time of the paralogous gene pairs suggested that the E2F/DP family expansion primarily occurred through a whole-genome duplication event. A regulatory element and coexpression network analysis indicated that E2F/DP regulated the expression of cell cycle-related genes. A yeast two-hybrid assay and expression analysis based on transcriptome data and in situ hybridization indicated that the PheE2F-PheDP complex played important roles in winter Moso bamboo shoot growth. The qRT-PCR results showed that the PheE2F/DPs exhibited diverse expression patterns in response to drought and salt treatment and diurnal cycles. CONCLUSION: Our findings provide novel insights into the Moso bamboo E2F/DP family and partial experimental evidence for further functional verification of the PheE2F/DPs.

Prevalence, predictors, and mortality of bloodstream infections due to methicillin-resistant Staphylococcus aureus in patients with malignancy: systemic review and meta-analysis.

BACKGROUND: Cancer patients are more likely to develop and die of bloodstream infection (BSI) than noncancer patients. Methicillin-resistant Staphylococcus aureus (MRSA), which is associated with immense mortality and economic burden worldwide, is not covered by the recommended initial antibiotic therapy for cancer patients with BSI. This systemic review was performed to estimate the global methicillin-resistant Staphylococcus aureus (MRSA) prevalence among bacteremia in patients with malignancy, and further study the predictors and mortality of cancer patients with MRSA bacteremia. METHODS: The PubMed and EMBASE databases were searched for studies published from Jan. 2000 to Mar. 2020 that provided primary data on the prevalence, predictors, or mortality of MRSA bacteremia in cancer patients. A random-effects model meta-analysis was performed to estimate the pooled prevalence of MRSA with 95% confidence intervals (95% CIs). RESULTS: The pooled prevalence of MRSA was 3% (95% CI 2-5%) among all bloodstream infections (BSIs) and 44% (95% CI 32-57%) among S. aureus bacteremia in cancer patients. Based on geographical stratification, the pooled prevalence was 5% in Africa (95% CI 1-14%), 1% in Americas (95% CI 1-2%), 2% in Europe (95% CI 1-4%), 4% in Western Pacific (95% CI 2-7%), 8% in South-east Asia (95% CI 4-14%) and 0% in Eastern Mediterranean (95% CI 0-3%). No significant temporal change in MRSA rates was detected in this analysis (R2 = 0.06; P = 0.24). Predictors for MRSA BSIs among cancer patients were identified by comparison with their methicillin-susceptible counterparts, and they were mainly related to healthcare-associated infections and immunosuppression. Finally, the 60-day mortality in adult cancer patients with MRSA BSIs was reported to be 12%, and the 6-month overall mortality was 43.2%, with community-onset infection, secondary BSI, and vancomycin MIC≥2 g/mL being the risk factors for mortality. CONCLUSIONS: Although the prevalence of MRSA BSIs among cancer patients is relatively low, it did not decline over time as MRSA BSIs in the general hospital population and the high mortality rate was related to MRSA BSIs in patients with malignancy.

Transcriptome analysis of terpenoid biosynthetic genes and simple sequence repeat marker screening in Eucommia ulmoides.

Trans-polyisoprene rubber is produced in the tissues of leaves, bark, and fruit of Eucommia ulmoides and is considered an important energy source. Transcript profiles of two tissues from E. ulmoides cv. Qinzhong No. 3, leaf and fruit, were analysed using the Illumina HiSeq 2000 system. In total, 104 million clean reads were obtained and assembled into 58,863 unigenes. Through gene functional classification, 28,091 unigenes (47.72%) were annotated and 65 unigenes have been hypothesized to encode proteins involved in terpenoid biosynthesis. In addition, 10,041 unigenes were detected as differentially expressed unigenes, and 29 of them were putatively related to terpenoid biosynthesis. The synthesis of trans-polyisoprene rubbers in E. ulmoides was hypothesised to be dominated by the mevalonate pathway. Farnesyl diphosphate synthase 2 (FPPS2) was considered a key component in the biosynthesis of trans-polyprenyl diphosphate. Rubber elongation factor 3 (REF3) might be involved in stabilising the membrane of rubber particles in E. ulmoides. To date, 351 simple sequence repeats (SSRs) were validated as polymorphisms from eight E. ulmoides plants (two parent plants and six F1 individuals), and these could act as molecular markers for genetic map density increase and breeding improvement of E. ulmoides.

Chemical composition and antioxidant activity of essential oil of six pinus taxa native to China.

The essential oils obtained by steam distillation from needles of six China endemic Pinus taxa (P. tabulaeformis, P. tabulaeformis f. shekanensis, P. tabulaeformis var. mukdensis, P. tabulaeformis var. umbraculifera, P. henryi and P. massoniana) were analysed by GC/MS. A total of 72 components were separated and identified by GC/MS from the six taxa. The major constituents of the essential oils were: α-pinene (6.78%-20.55%), bornyl acetale (3.32%-12.71%), ß-caryophellene (18.26%-26.31%), α-guaiene (1.23%-8.19%), and germacrene D (1.26%-9.93%). Moreover, the essential oils were evaluated for antioxidant potential by three assays (DPPH, FRAP and ABTS) and tested for their total phenolic content. The results showed that all essential oils exhibited acceptable antioxidant activities and these strongly suggest that these pine needles may serve as a potential source of natural antioxidants for food and medical purposes.

A molecular genetic linkage map of Eucommia ulmoides and quantitative trait loci (QTL) analysis for growth traits.

Eucommia ulmoides is an economically important tree species for both herbal medicine and organic chemical industry. Effort to breed varieties with improved yield and quality is limited by the lack of knowledge on the genetic basis of the traits. A genetic linkage map of E. ulmoides was constructed from a full-sib family using sequence-related amplified polymorphism, amplified fragment length polymorphism, inter-simple sequence repeat and simple sequence repeat markers. In total, 706 markers were mapped in 25 linkage groups covering 2133 cM. The genetic linkage map covered approximately 89% of the estimated E. ulmoides genome with an average of 3.1 cM between adjacent markers. The present genetic linkage map was used to identify quantitative trait loci (QTL) affecting growth-related traits. Eighteen QTLs were found to explain 12.4%-33.3% of the phenotypic variance. This genetic linkage map provides a tool for marker-assisted selection and for studies of genome in E. ulmoides.

Phenotypic Trait Subdivision Provides New Sight Into the Directional Improvement of Eucommia ulmoides Oliver.

Eucommia ulmoides Oliver has been used extensively in many fields. To satisfy increasing demand, great efforts must be made to further improve its traits. However, limited information is available on these traits, which is a factor that restricts their improvement. To improve traits directionally, nine clones were assigned to six sites to analyze the effect of different variation sources (the genotype, site, and genotype × environment interaction) on the phenotypic trait. In addition, a mixed linear model was used to assess the contribution of variations. In general, for most traits, the site effect accounted for a larger proportion of the variance, followed by the genotype and genotype × environment interaction effects. All the studied genotypes and sites had a significant effect, indicating that they could be improved by selecting preferable genotypes or cultivation areas, respectively. Interestingly, growth traits or economic traits could be improved simultaneously. Trait performance and stability are necessary when selecting genotypes. Moreover, the discriminating ability of genotypes should be considered in selecting cultivation areas. Annual mean temperature and annual sunshine duration proved to be crucial factors that affected the traits. They were correlated positively with economic traits and leaf yield and correlated negatively with growth traits. These findings contributed to selecting a wider range of cultivation areas. Regarding the genotype × environment interaction effect, there were significant differences only in the gutta-percha content, the total number of leaves, and the chlorogenic acid content. These traits could also be improved by choosing appropriate genotypes for the local environment. The research has provided preliminary data on the main factors that affect the traits of E. ulmoides and offered solutions for trait improvement. This information could be a reference for the trait improvement of other plants.

Identification of a male-specific Amplified Fragment Length Polymorphism (AFLP) and a Sequence Characterized Amplified Region (SCAR) marker in Eucommia ulmoides Oliv.

Eucommia ulmoides Oliv. is a dioecious species. Currently, there is no method to identify the sex during the juvenile stage that lasts a relatively long time. This study aimed to develop a sex specific Amplified Fragment Length Polymorphism (AFLP) marker for Eucommia ulmoides Oliv. Of a total of 64 AFLP primer combinations screened, primer combination E-ACA/M-CTT produced a 350 bp male-specific marker. This 350 bp AFLP marker was converted into a 247 bp Sequence Characterized Amplified Region (SCAR) marker. Results suggest that the SCAR marker can be utilized for early sexual identification in Eucommia ulmoides Oliv., and it will greatly facilitate future breeding programs.

Trait Variations and Probability Grading Index System on Leaf-Related Traits of Eucommia ulmoides Oliver Germplasm.

Eucommia ulmoides Oliver (EUO), an economic tree grown specifically in China, is widely used in various fields. To satisfy the requirements of industrial development, superior varieties need to be selected for different uses. However, there is no unified standard for breeders to reference. In this study, leaf-related traits were classified by a probability grading method. The results indicated there were significant differences between different planting models for the studied traits, and the traits in the Arbor forest model showed more abundant variation. Compared with genotype, the planting model accounted for relatively bigger variance, indicating that the standard should be divided according to planting models. Furthermore, the optimum planting model for different traits would be obtained by analyzing the variation range. Association analyses were conducted among traits to select the crucial evaluation indexes. The indexes were divided into three grades in different planting models. The evaluation system on leaf-related traits of EUO germplasm was established preliminarily, which considered planting models and stability across years for the first time. It can be treated as a reference to identify and evaluate EUO germplasm resources. Additionally, the study served as an example for the classification of quantitative traits in other economically important perennial plants.

A Promising Future of Ferroptosis in Tumor Therapy.

Currently, mechanisms and therapeutic approaches have been thoroughly studied in various prevalent malignant tumors, such as breast and lung cancer. However, there is inevitable tumor progression and drug resistance. Uncovering novel treatment strategies to inhibit tumor development is important. Ferroptosis, a form of cell death associated with iron and lipid peroxidation, has drawn extensive attention. In this paper, we reviewed the underlying mechanisms of ferroptosis (i.e., iron, glutathione, and lipid metabolism) and its role in various tumors (i.e., lung cancer, liver carcinoma, breast cancer, and pancreatic cancer). Moreover, we summarized ferroptosis-related anti-tumor drugs and emphasized the potential of combined treatment of anti-tumor drugs and radiotherapy in an effort to provide novel anti-tumor treatments.

Characterization of diacylglycerol acyltransferase 2 from Idesia polycarpa and function analysis.

Idesia polycarpa is an oil-producing tree native to China and Northeast Asia. The fruits of I. polycarpa which are named oil grape are unique in that they contain large amounts saturated and unsaturated lipids. Diacylglycerol acyltransferase 2 (DGAT2) is a key enzyme catalyzing the final step of triacylglyceride (TAG) synthesis. However, expression and bioinformatics of DGAT2 in I. polycarpa are still blank. In order to further understand the lipogenesis of oil grape, we contrasted seven various growth periods fruits from seed formation to seed maturation. Lipid accumulation rates and final lipid content were significantly different among the different periods. We cloned and characterized the DGAT2 gene from fruits of I. polycarpa. A partial fragment of 239 bp of IpDGAT2 was amplified by PCR. We cloned the open-reading frame (ORF) of IpDGAT2 by RACE technique. The ORF of IpDGAT2 contains 984 bp and encodes 327 amino acids. The qPCR analysis manifested that IpDGAT2 was expressed in all oil grape growing periods and expression was highest on September 20 (seed maturation). In I. polycarpa fruits the expression of IpDGAT2 was positively correlated with the lipid accumulation rates. Rhodotorula glutinis expression analysis showed that IpDGAT2 have a diacylglycerol acyltransferase bio-functional. Heterologous expression of the 35S::IpDGAT2 in Arabidopsis thaliana confirmed that the isolated IpDGAT2 could catalyze lipid synthesis. The lipid content increased by 40 % in transgenic plants relative to the control. which suggests that high lipid content fruits can be created by the overexpression of IpDGAT2 in I. polycarpa.

MicroRNA-30a targets BECLIN-1 to inactivate autophagy and sensitizes gastrointestinal stromal tumor cells to imatinib.

Gastrointestinal stromal tumors (GISTs), the most widespread type of sarcoma, contain driver gene mutations predominantly of receptor tyrosine kinase and platelet-derived growth factor receptor alpha. However, the inevitable development of resistance to imatinib (IM) cannot be fully attributed to secondary driver gene mutations. In this study, we investigated the role of microRNA-30a in sensitization of GIST cells to IM in vivo and in vitro. Higher levels of miR-30a were detected in GIST-T1 cells, which were more sensitive to IM than GIST-882 cells. IM treatment also reduced miR-30a levels, indicating the possible role of miR-30a in GIST IM resistance. Subsequently, miR-30a was confirmed to be an IM sensitizer via a mechanism that was attributed to its involvement in the regulation of cell autophagy. The interaction of miR-30a and autophagy in IM treated GIST cells was found to be linked by beclin-1. Beclin-1 knockdown increased IM sensitivity in GIST cell lines. Finally, miR-30a was confirmed to enhance IM sensitivity of GIST cells in mouse tumor models. Our study provides evidence for the possible role of miR-30a in the emergence of secondary IM resistance in GIST patients, indicating a promising target for overcoming this chemoresistance.

Denosumab and Sunitinib in the treatment of giant-cell tumor of bone with pulmonary and bone metastases in an adolescent: A case report.

INTRODUCTION: Giant cell tumor of bone with pulmonary and bone metastases is exceedingly rare in adolescents. Furthermore, Denosumab and Sunitinib in the treatment of giant cell tumor of bone has never been reported. PATIENT CONCERNS: A 16-year-old boy complained of fever, chest tightness, and shortness of breath and back pain for 5 days. DIAGNOSIS: Giant cell tumor of bone with pulmonary and bone metastases. INTERVENTIONS: The patient underwent 2 surgeries for giant cell tumor of bone located in the spine and received Denosumab to reduce local recurrence and control the metastases. Then Sunitinib was added into treatment strategies after the progressing of metastases. OUTCOMES: Within 5 months of Denosumab and Sunitinib, lung metastases were shrinking (stable disease, response evaluation criteria in solid tumors version 1.1). Until now about 4 years into treatment the patient is still survival. Pulmonary and bone metastases are stable. CONCLUSIONS: This is a case of multi-center giant cell tumor of bone, it does not only provide a reference to the treatment of similar cases of the clinic but also reflects the limitations of the application of Denosumab in the real world.

Molecular cloning and function analysis of FAD2 gene in Idesia polycarpa.

Idesia polycarpa is a valuable oil-producing tree and can potentially be used for edible oil and biofuel production. The fruits of I. polycarpa are unique in that they contain both saturated and unsaturated lipids. Fatty acid desaturase 2 (FAD2), also as known as omega-6 fatty acid desaturase in endoplasmic, is a key enzyme for linoleic acid and α-linolenic acid biosynthesis. However, bioinformatics and expression of FAD2 in I. polycarpa are still absent. Here, to gain insight into the lipid and linoleic synthesis of I. polycarpa, we compared the fruits from different growth stages. Lipid accumulation rates, final lipid content, linoleic accumulation rates and final linoleic content were significantly different among the different stages. In a further step, the FAD2 gene from fruits of I. polycarpa, named IpFAD2, was cloned and characterized. A partial fragment of 169 bp of IpFAD2 was amplified by degenerate PCR. Full cDNA of IpFAD2 was obtained by the RACE technique. The open-reading frame of IpFAD2 was 1149 bp in length, encoding 382 amino acids. A comparison of the deduced amino acids sequence of IpFAD2 with FAD2 from other species showed high similarities, ranging from 78.8 to 92.6%. The IpFAD2-predicted protein has a theoretical molecular mass of 44.03 kDa and an isoelectric point (pI) of 8.04. It has five transmembrane helices located on the endoplasmic reticulum. The IpFAD2-predicted protein was classified as belonging to the Membrane-FADS-like superfamily based on its conserved domain analysis. Expression analysis based on qRT-PCR indicated that IpFAD2 was expressed in different fruit growth stages, with the highest expression level at 80 DAP and the lowest at 130 DAP. The expression of IpFAD2 was positively correlated with the linoleic accumulation rates in I. polycarpa fruits. Prokaryotic expression in Escherichia. Coli BL21(DE3) indicated that IpFAD2 gene could encode a bio-functional omega-6 fatty acid desaturase. Heterologous expression in Arabidopsis thaliana confirmed that the isolated IpFAD2 proteins could catalyse linoleic synthesis. This is the first cloning and expression analysis of FAD2 from I. polycarpa, significantly contributing to our understanding of the role of IpFAD2 in linoleic synthesis, esp. in terms of genetic engineering breeding for linoleic production.

Selection of suitable reference genes for qRT-PCR normalisation under different experimental conditions in Eucommia ulmoides Oliv.

Normalisation of data, by choosing the appropriate reference genes, is fundamental for obtaining reliable results in quantitative real-time PCR (qPCR). This study evaluated the expression stability of 11 candidate reference genes with different varieties, developmental periods, tissues, and abiotic stresses by using four statistical algorithms: geNorm, NormFinder, BestKeeper, and RefFinder. The results indicated that ubiquitin-conjugating enzyme S (UBC) and ubiquitin-conjugating enzyme E2 (UBC E2) could be used as reference genes for different E. ulmoides varieties and tissues, UBC and histone H4 (HIS4) for different developmental periods, beta-tubulin (TUB) and UBC for cold treatment, ubiquitin extension protein (UBA80) and HIS4 for drought treatment, and ubiquitin-60S ribosomal protein L40 (UBA52) and UBC E2 for salinity treatment. UBC and UBC E2 for the group "Natural growth" and "Total", UBA80 and UBC for the group "Abiotic stresses". To validate the suitability of the selected reference genes in this study, mevalonate kinase (MK), phenylalanine ammonia-lyase (PAL), and 4-coumarate-CoA ligase (4CL) gene expression patterns were analysed. When the most unstable reference genes were used for normalisation, the expression patterns had significant biases compared with the optimum reference gene combinations. These results will be beneficial for more accurate quantification of gene expression levels in E. ulmoides.

The first genetic linkage map of Eucommia ulmoides.

In accordance with pseudo-testcross strategy, the first genetic linkage map of Eucommia ulmoides Oliv. was constructed by an F1 population of 122 plants using amplified fragment length polymorphism (AFLP) markers. A total of 22 AFLP primer combinations generated 363 polymorphic markers. We selected 289 markers segregating as 1:1 and used them for constructing the parent-specific linkage maps. Among the candidate markers, 127 markers were placed on the maternal map LF and 108 markers on the paternal map Q1. The maternal map LF spanned 1116.1 cM in 14 linkage groups with a mean map distance of 8.78 cM; the paternal map Q1 spanned 929.6 cM in 12 linkage groups with an average spacing of 8.61 cM. The estimated coverage of the genome through two methods was 78.5 and 73.9% for LF, and 76.8 and 71.2% for Q1, respectively. This map is the first linkage map of E. ulmoides and provides a basis for mapping quantitative-trait loci and breeding applications.