Whole-genome resequencing using next-generation and Nanopore sequencing for molecular characterization of T-DNA integration in transgenic poplar 741.

BACKGROUND: The molecular characterization information of T-DNA integration is not only required by public risk assessors and regulators, but is also closely related to the expression of exogenous and endogenous genes. At present, with the development of sequencing technology, whole-genome resequencing has become an attractive approach to identify unknown genetically modified events and characterise T-DNA integration events. RESULTS: In this study, we performed genome resequencing of Pb29, a transgenic high-resistance poplar 741 line that has been commercialized, using next-generation and Nanopore sequencing. The results revealed that there are two T-DNA insertion sites, located at 9,283,905-9,283,937 bp on chromosome 3 (Chr03) and 10,868,777-10,868,803 bp on Chr10. The accuracy of the T-DNA insertion locations and directions was verified using polymerase chain reaction amplification. Through sequence alignment, different degrees of base deletions were detected on the T-DNA left and right border sequences, and in the flanking sequences of the insertion sites. An unknown fragment was inserted between the Chr03 insertion site and the right flanking sequence, but the Pb29 genome did not undergo chromosomal rearrangement. It is worth noting that we did not detect the API gene in the Pb29 genome, indicating that Pb29 is a transgenic line containing only the BtCry1AC gene. On Chr03, the insertion of T-DNA disrupted a gene encoding TAF12 protein, but the transcriptional abundance of this gene did not change significantly in the leaves of Pb29. Additionally, except for the gene located closest to the T-DNA integration site, the expression levels of four other neighboring genes did not change significantly in the leaves of Pb29. CONCLUSIONS: This study provides molecular characterization information of T-DNA integration in transgenic poplar 741 line Pb29, which contribute to safety supervision and further extensive commercial planting of transgenic poplar 741.

Combination of NGAL and Cystatin C for Prediction of Preeclampsia at 10-14 Weeks of Gestation.

BACKGROUND: Preeclampsia (PE) is a severe pregnancy complication and is an important cause for maternal and child death, premature delivery, and limited intrauterine growth and development. The aim of this study was to investigate the role of NGAL and cystatin C, alone and in combination, for early prediction of PE at 10 - 14 weeks of gestation. METHODS: Serum levels of NGAL and cystatin C were assessed in women at 10 - 14 weeks of gestation who subsequently developed PE (n = 128) and normal pregnancy outcome (n = 183). Comparison of clinical characteristics, NGAL, and cystatin C levels between normal pregnancy and PE groups were analyzed using Mann-Whitney test. The receiver operating characteristic curve (ROC curve) was used to analyze the value of serum NGAL and cystatin C levels in predicting PE. RESULTS: The levels of cystatin C and NGAL in the serum were significantly higher in the PE group [0.64 mg/L (0.52 - 0.78)] and [34.9 ng/mL (24.4 - 55.2), respectively] than in the normal pregnancy group [0.56 mg/L (0.49 - 0.65)] and [20.2 ng/mL (13.8 - 26.9), respectively]. ROC curve analysis showed that serum NGAL levels predicted the area under the curve in the PE period 0.739 (95% CI: 0.618 to 0.860). Serum cystatin C levels predicted the area under the curve in the PE period 0.722 (95% CI: 0.592 to 0.853). The combination of serum NGAL and cystatin C levels predicted the area under the curve in the PE period 0.877 (95% CI: 0.811 to 0.943). CONCLUSIONS: NGAL and cystatin C levels in serum appear to be ideal biomarkers for PE prediction at 10 - 14 weeks. The combination of NGAL and cystatin C will also be more valuable in discriminating patients at risk of developing PE from other pregnancy complications early in gestation.

Development of taxane-containing chemotherapy of doxorubicin-resistant diffuse large B-cell lymphoma.

Recurrent diffuse large B-cell lymphoma (DLBCL) presents a significant challenge in the clinic, which necessitates the development of alternative therapeutic regimens. Here, we quantified the growth inhibition rate of a panel of clinically approved anticancer drugs in a patient-derived DLBCL cell line that is resistant to doxorubicin, a central component of the standard of care R-CHOP. Our study indicates that this cell line is especially sensitive to two taxanes, docetaxel and paclitaxel. Furthermore, the effective dose of taxane can be lowered when it is combined with other anticancer drugs. Our study provides guidance for preclinical studies to test alternative regimens for recurrent DLBCL.

Downregulation of circAsxl2 Relieves Neuronal Injury Induced by oxygen-glucose deprivation/reperfusion.

BACKGROUND: Circular RNAs (circRNAs) have been shown to play an important role in cerebral ischemia-reperfusion (I/R) injury. However, the role of circAsxl2 (mmu_circ_0000346) in cerebral I/R injury remains unclear. METHODS: Mouse brain neuronal cell line (HT-22) was used to perform oxygen-glucose deprivation/reperfusion (OGD/R) treatment. The levels of circAsxl2, microRNA (miR)-130b-5p and forkhead box O3 (FOXO3) were determined using quantitative real-time PCR. Cell viability and apoptosis were measured using cell counting kit 8 assay and flow cytometry. Commercial kits were used to assess cell cytotoxicity, inflammation and oxidative stress. Protein expression was analyzed by western blot. RNA interaction was verified using dual-luciferase reporter assay, RIP assay and RNA pull-down assay. RESULTS: CircAsxl2 was highly expressed in OGD/R-induced HT-22 cells, and its silencing could alleviate OGD/R-induced apoptosis, inflammation and oxidative stress in HT-22 cells. MiR-130b-5p was sponged by circAsxl2, and its inhibitor could overturn the regulation of circAsxl2 knockdown on OGD/R-induced neuronal injury. FOXO3 was targeted by miR-130b-5p and its expression was positively regulated by circAsxl2. In addition, the regulation of circAsxl2 knockdown on OGD/R-induced neuronal injury also was reversed by FOXO3 overexpression. CONCLUSION: CircAsxl2/miR-130b-5p/FOXO3 axis accelerated OGD/R-induced neuronal injury, which might provide effective strategies for treating cerebral I/R injury.

Integrated physiological, transcriptomic, and metabolomic analyses of drought stress alleviation in Ehretia macrophylla Wall. seedlings by SiO2 NPs (silica nanoparticles).

With environmental problems such as climate global warming, drought has become one of the major stress factors, because it severely affects the plant growth and development. Silicon dioxide nanoparticles (SiO2 NPs) are crucial for mitigating abiotic stresses suffered by plants in unfavorable environmental conditions and further promoting plant growth, such as drought. This study aimed to investigate the effect of different concentrations of SiO2 NPs on the growth of the Ehretia macrophylla Wall. seedlings under severe drought stress (water content in soil, 30-35%). The treatment was started by starting spraying different concentrations of SiO2 NPs on seedlings of Ehretia macrophyla, which were consistently under normal and severe drought conditions (soil moisture content 30-35%), respectively, at the seedling stage, followed by physiological and biochemical measurements, transcriptomics and metabolomics analyses. SiO2 NPs (100 mg·L-1) treatment reduced malondialdehyde and hydrogen peroxide content and enhanced the activity of antioxidant enzymes under drought stress. Transcriptomic analysis showed that 1451 differentially expressed genes (DEGs) in the leaves of E. macrophylla seedlings were regulated by SiO2 NPs under drought stress, and these genes mainly participate in auxin signal transduction and mitogen-activated protein kinase signaling pathways. This study also found that the metabolism of fatty acids and α-linolenic acids may play a key role in the enhancement of drought tolerance in SiO2 NP-treated E. macrophylla seedlings. Metabolomics studies indicated that the accumulation level of secondary metabolites related to drought tolerance was higher after SiO2 NPs treatment. This study revealed insights into the physiological mechanisms induced by SiO2 NPs for enhancing the drought tolerance of plants.

Effects of Carvedilol on Blood Pressure, Blood Sugar, and Blood Lipids in Elderly Patients with Refractory Hypertension.

Refractory hypertension seriously affects the life safety of patients. To investigate the effect of carvedilol combined with conventional antihypertensive therapy on blood pressure, blood sugar, blood lipids, and cardiovascular and cerebrovascular complications in elderly patients with refractory hypertension, a total of 80 elderly patients with refractory hypertension who were admitted from June 2019 to September 2021 were selected as the retrospective research objects and divided into the observation group and the control group according to the random number table method, 40 cases in each group, and the control group received conventional antihypertensive therapy. On this basis, the observation group was treated with carvedilol, and the effects of blood pressure, blood sugar, blood lipids, and cardiovascular and cerebrovascular complications were analyzed in the two groups. After treatment, the systolic blood pressure, diastolic blood pressure, adrenal medulla (AM), and endothelin (ET) of the observation group were significantly lower than those of the control group, but the heart rate and NO of the observation group were significantly higher than those of the control group. After treatment, the fasting blood glucose, insulin, triacylglycerol (TG), cholesterol (CHO), and low-density lipoprotein (LDL-C) of the observation group were significantly higher than those of the control group, but the ISI and HDL-C of the observation group were significantly lower than those of the control group. After treatment, the levels of IL-1ß, IL-6, TNF-α, UAER, BUN, and SCr in the observation group were significantly lower than those in the control group. The TC of the observation group was lower than that of the control group. Combination therapy has a remarkably stable, sustained, and safe antihypertensive effect on patients with refractory hypertension.

Effects of three regeneration methods on the growth and bacterial community diversity of Populus × euramericana.

To study the effects of different regeneration methods on the growth and bacterial community diversity of Populus × euramericana cv. '74/76' (poplar 107), we investigated the growth of poplar 107 trees under three regeneration methods in 2017 and 2020, and sequenced the 16S rDNA V5-V7 regions in stem endophytic, root endophytic, and rhizosphere soil bacteria present in samples from the three regeneration methods using the Illumina high-throughput sequencing platform. The growth analysis showed that stump grafting regeneration (ST) and stump sprouting regeneration (SP) presented similar tree height and diameter at breast height (DBH), which were significantly lower by planted seedling regeneration (CK). The high-throughput sequencing results showed that the rhizosphere soil bacteria appeared to be significantly more diverse and rich than the root and stem endophytic bacteria. Cluster analysis showed that the similarity of bacterial community structure among the rhizosphere soil, root, and stem was small. Thus, the three sample types showed significant differences in bacteria. While comparing the two years, 2020 was significantly more diverse and rich than 2017. With the increase in stand age, the abundance of Proteobacteria increased and the abundance of Acidobacteria decreased. Among the three regeneration methods, ST significantly increased the diversity of stem endophytic bacteria. Chthoniobacter was enriched in SP, which promoted the decomposition of organic matter, and more plant growth promoting rhizobacteria (PGPR) were accumulated in the rhizosphere of SP and ST. The composition of the bacterial community was similar in the three regeneration methods, but the community composition was different. Regeneration and transformation of poplar plantations can be better carried out by stump grafting and stump sprouting.

Correlation Analysis of the Bacterial Community and Wood Properties of Populus × euramericana cv. "74/76" Wet Heartwood.

Wetwood disease of poplar limits the processing and manufacturing of poplar, and the pathogenic bacteria of wet heartwood are poorly known. We used high-throughput sequencing methods to analyze the bacterial community of the heartwood, sapwood, root tissue, and rhizosphere soil of Populus × euramericana cv. "74/76" (poplar 107) in wetwood trees and healthy trees to explore the cause of poplar wetwood disease. Bacterial diversity and community structure were analyzed, and the correlation between wood properties and bacterial relative abundance was analyzed to explore their relationship. Two alpha-diversity indices of endophytic bacteria in the heartwood of wetwood trees were significantly (p < 0.05) lower than that in the heartwood of healthy trees, and the community structure between the two types of trees was significantly different. No significant differences in the alpha-diversity indices nor community structure were observed in the sapwood, root tissue, or rhizosphere bacterial community of diseased and healthy trees. The distribution of dominant bacteria genus in the heartwood of diseased and healthy trees differed. Proteiniphilum, Actinotalea, and Methanobacterium were the dominant genera in diseased trees' heartwood. Proteiniphilum, Dysgonomonas, and Bacteroides were the dominant genera in healthy trees' heartwood. The relative abundance of Proteiniphilum, Actinotalea, and Methanobacterium was significantly higher in the heartwood of wetwood trees than those of healthy trees. A db-RDA analysis found that these three bacterial genera were positively correlated with the rate of wet heartwood. These three bacterial genera may be the main pathogens causing poplar wetwood disease.

The effect of vitamin D on the lipid profile as a risk factor for coronary heart disease in postmenopausal women: a meta-analysis and systematic review of randomized controlled trials.

BACKGROUND AND AIM: The exact effect of vitamin D administration on the lipid profile in postmenopausal women is unknown. However, as dyslipidemia is a recognized risk factor for coronary heart disease (CHD) in this population, the lipid-lowering effects of vitamin D need to be explored Thus, we conducted a systematic review and meta-analysis of randomized controlled trials (RCTs) that evaluated the impact of vitamin D use on triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and total cholesterol (TC) as a risk factor for coronary heart disease (CHD) in postmenopausal women. METHODS: We developed a search strategy for multiple databases (PubMed/Medline, Scopus, Embase, and Web of Science) to identify relevant RCTs whose results were published until June 1st, 2021. We combined the results using a random effects model (the DerSimonian and Laird random effects model). Lipid profile outcomes were expressed as weighted mean difference (WMD) and 95% confidence intervals (CIs) between intervention and comparator groups. RESULTS: Supplementation with vitamin D decreased TG (WMD: -3.55 mg/dL, 95% CI: -5.34 to -1.76, P < 0.001) in postmenopausal females versus controls. In the subgroup analyses, vitamin D increased TC when the treatment duration was ˂26 weeks (WMD: 6.56 mg/dL, 95% CI: 0.78 to 12.35, P = 0.02) as compared to ≥26 weeks (WMD: -2.06 mg/dL, 95% CI: -5.49, 1.36, P = 0.23) and in the participants with a BMI ≥30 kg/m2 (WMD: 3.65 mg/dL, 95% CI: 0.09, 7.22, P = 0.044). Moreover, vitamin D increased HDL-C when the treatment duration was ˂26 weeks (WMD: 2.67 mg/dL, 95% CI: 0.66 to 4.68, P = 0.009). In addition, vitamin D decreased LDL-C when the vitamin D dose was ˃400 IU/day (WMD: -1.89 mg/dL, 95% CI: -2.47 to -1.31, P < 0.001) as compared to ≤400 IU/day (WMD: 2.50 mg/dL, 95% CI: -2.50, 7.52, P = 0.327). CONCLUSIONS: Vitamin D administration on the lipid profile as a risk factor for CHD in postmenopausal women reduces TG. Its effects to lower LDL-C and increase HDL-C and TC levels are clinically negligible but should be investigated in future research. In addition, supplementation with vitamin D results in a clinically significant reduction in TG, particularly in postmenopausal females with hypertriglyceridemia at baseline.

MicroRNA-204 targets signal transducer and activator of transcription 5 expression and inhibits proliferation of B-cell lymphoma cells.

The function of microRNAs (miRNAs) in tumorigenesis has been extensively investigated. In the present study, the aim was to investigate the expression and role of miR­204 in B­cell lymphoma. The present study demonstrated that miR­204 is downregulated in B­cell lymphoma. Using in vitro studies, overexpression of miR­204 was shown to inhibit growth in Daudi and Raji B­cell lymphoma cell lines. Furthermore, miR­204 could bind the 3'­untranslated region of signal transducer activator of transcription 5 (STAT5), a transcription factor that promotes B­cell lymphoma oncogenesis. Re­introduction of STAT5 reversed the antiproliferative roles of miR­204, confirming the specific importance of STAT5 for miR­204 action in cell proliferation. The present study suggests a novel mechanism for dysregulated miRNAs in the progression of B­cell lymphoma.

Human cytomegalovirus-encoded miR-US25-1 aggravates the oxidised low density lipoprotein-induced apoptosis of endothelial cells.

Human cytomegalovirus (HCMV) infection is linked to the development and severity of the cardiovascular disease atherosclerosis; however, there is little known about the promotion of atherosclerosis. miR-US25-1 is one of HCMV-encoded miRNAs and targets cellular genes that are essential for virus growth to control the life cycle of the virus and host cells. The prominent regulation on cell cycle genes of the miR-US25-1 attracts us to explore its role in the atherosclerosis promotion. It was indicated that miR-US25-1 level was upregulated in subjects or in endothelial cells with HCMV infection; and the miR-US25-1 downregulated the expression of BRCC 3 by targeting the 5' UTR of BRCC 3. And a miR-US25-1 mimics transfection could reduce the EAhy926 cell viability but did not induce apoptosis in EAhy926 cells. And what is more, miR-US25-1 mimicis transfection deteriorated the ox-LDL-induced apoptosis and aggravated the upregulation of apoptosis-associated molecules by oxidised low density lipoprotein (ox-LDL) in EAhy926 cells. And we have also confirmed the deregulation of BRCC 3 expression by miR-US25-1 by targeting the 5' UTR of it. Given the vital role of BRCC 3 in DNA damage repairing, we speculated that the targeting inhibition of BRCC 3 by miR-US25-1 may contribute to the aggravation of ox-LDL-promoted apoptosis of endothelial EAhy926 cells.