Application of a novel strong promoter from Chinese fir (Cunninghamia lanceolate) in the CRISPR/Cas mediated genome editing of its protoplasts and transgenesis of rice and poplar

Novel constitutive promoters are essential for plant biotechnology. Although in angiosperms, a number of promoters were applied in monocots or dicots genetic engineering, only a few promoters were used in gymnosperm. Here we identified two strong promoters (Cula11 and Cula08) from Chinese fir (C. lanceolate) by screening the transcriptomic data and preliminary promoter activity assays in tobacco. By using the newly established Chinese fir protoplast transient expression technology that enables in vivo molecular biology studies in its homologous system, we compared the activities of Cula11 and Cula08 with that of the commonly used promoters in genetic engineering of monocots or dicots, such as CaM35S, CmYLCV, and ZmUbi, and our results revealed that Cula11 and Cula08 promoters have stronger activities in Chinese fir protoplasts. Furthermore, the vector containing Cas gene driven by Cula11 promoter and sgRNA driven by the newly isolated CulaU6b polyIII promoters were introduced into Chinese fir protoplasts, and CRISPR/Cas mediated gene knock-out event was successfully achieved. More importantly, compared with the commonly used promoters in the genetic engineering in angiosperms, Cula11 promoter has much stronger activity than CaM35S promoter in transgenic poplar, and ZmUbi promoter in transgenic rice, respectively, indicating its potential application in poplar and rice genetic engineering. Overall, the novel putative constitutive gene promoters reported here will have great potential application in gymnosperm and angiosperm biotechnology, and the transient gene expression system established here will serve as a useful tool for the molecular and genetic analyses of Chinese fir genes

Multifunctional Gelatin-Nanoparticle-Modified Chip for Enhanced Capture and Non-Destructive Release of Circulating Tumor Cells

Circulating tumor cells (CTCs) in cancer patients’ peripheral blood have been demonstrated to be a significant biomarker for metastasis detection, disease prognosis, and therapy response. Due to their extremely low concentrations, efficient enrichment and non-destructive release are needed. Herein, an FTO chip modified with multifunctional gelatin nanoparticles (GNPs) was designed for the specific capture and non-destructive release of CTCs. These nanoparticles share a similar dimension with the microvilli and pseudopodium of the cellular surface; thus, they can enhance adhesion to CTCs, and then GNPs can be degraded by the enzyme matrix metalloproteinase (MMP-9), gently releasing the captured cells. In addition, the transparency of the chip makes it possible for fluorescence immunoassay identification in situ under a microscope. Our chip attained a high capture efficiency of 89.27%, a release efficiency of 91.98%, and an excellent cellular viability of 96.91% when the concentration of MMP-9 was 0.2 mg/mL. Moreover, we successfully identified CTCs from cancer patients’ blood samples. This simple-to-operate, low-cost chip exhibits great potential for clinical application

Global Trends in Atherosclerosis Research in the Epigenetics Field: Bibliometric and Visualization Studies

Atherosclerosis is a pathological vascular state caused by the interaction of environmental and hereditary factors. Epigenetic modifications may be the bridge connecting environmental factors and genetic factors. A search for publications on the Web of Science database in the field of atherosclerosis related to epigenetics was conducted from the earliest mention to 31 December 2020. Data on total and annual publications, citations, impact factors, Hirsch (H)-index, citation times, most prolific authors, and frequently published journals were collected for quantitative and qualitative comparison. A total of 1848 publications related to epigenetics and atherosclerosis were found. The major contributing countries were the China (522, 28.23%), United States (485, 26.23%), and Germany (119, 6.44%). The greatest number of retrieved publications were published in the journal, “Arteriosclerosis, Thrombosis, and Vascular Biology” (62, 3.66%). The publication “Oxidative Stress and Diabetic Complications” was cited 2370 times. The most frequent keywords were “DNA methylation” and “LncRNA”. Publications on epigenetic research in the atherosclerosis field have increased significantly every year, indicating that the study of epigenetic modifications plays an increasingly important role in understanding the pathology of atherosclerosis

Positively Charged Polyprodrug Amphiphiles with Enhanced Drug Loading and Reactive Oxygen Species-Responsive Release Ability for Traceable Synergistic Therapy

Due to the vast differences in chemical properties among small molecule drugs, nucleotide drugs, and superparamagnetic iron oxide nanocubes (SPIONs), such as charge and hydrophobicity, entrapment of these within a single carrier for traceable synergistic therapy has been proven difficult. Herein, we synthesize positively charged polyprodrug amphiphiles. The hydrophobic polyprodrug unit of the amphiphiles is positively charged, which can simultaneously load hydrophobic SPIONs and absorb negative let-7b antisense oligonucleotide to construct traceable co-delivery nanoparticles (NPs). This characteristic avoids the use of inert materials and enhances drug loading of the traceable NPs. The traceable NPs can achieve controlled release of drugs to reduce the differentiation of exogenous neural stem cells (NSCs) and enhance their secretion of brain-derived neurotrophic factor (BDNF) synergistically. Exogenous NSCs treated with the NPs significantly rescue the memory deficits in 2xTg-AD mice. In addition, the transplantation site and migration of exogenous NSCs can be traced using the SPIONs with high r(2) value for magnetic resonance imaging. Therefore, traceable NPs self-assembled from the positively charged polyprodrug amphiphiles may have the potential to open up a new avenue for treatment of Alzheimer&#39;s disease (AD), as well as other neurodegenerative disorders.</p

Does Dietary Lipid Level Affect the Quality of Triploid Rainbow Trout and How Should It Be Assessed?

Organoleptic properties and nutritional value are the most important characteristics of fish fillet quality, which can be determined by a series of quality evaluation indexes and closely related to fish nutrition. Systematic organoleptic and nutritional quality evaluation indexes consisting of 139 indexes for physical properties and chemical compositions of triploid rainbow trout were established. Besides, effects of dietary lipid levels (6.6%, 14.8%, 22.8% and 29.4%) on the quality of triploid rainbow trout were analyzed in the study. The main results showed that, for fillet appearance quality, fish fed diets with lipid levels above 22.8% had higher fillet thickness and redness but lower gutted yield and fillet yield (p p p p < 0.05). For nutritional value, a high lipid diet could increase the lipid nutrition level (such as the content of long chain polyunsaturated fatty acids increased from 3.47 to 4.41 g/kg muscle) but decease tryptophan and selenium content (from 2.48 to 1.60 g/kg muscle and from 0.17 to 0.11 g/kg muscle, respectively). In total, a high lipid diet could improve the quality of triploid rainbow trout. The minimum dietary lipid level for triploid rainbow trout should be 22.8% to keep the better organoleptic and nutritional quality

Comparative study on the fillet nutritional quality of diploid and triploid rainbow trout (Oncorhynchus mykiss)

This study was conducted to investigate the fillet nutritional quality difference of diploid and triploid rainbow trout with the same genetic background, farming condition and body weight. The results showed that, 1) diploid rainbow trout had significantly higher content of protein, valine, cysteine, histidine and proline, and higher essential amino acid index, nutrition index and biological value than triploid rainbow trout (P  0.05), which indicated that both diploid and triploid rainbow trout fillets were balanced and suitable for human consumption. 2) Triploid rainbow trout had significantly higher content of lipid, C18:1n-9, C18:2n-6, C20:5n-3, C22:6n-3, unsaturated fatty acids (UFA), polyunsaturated fatty acids (PUFA), long chain polyunsaturated fatty acids, n-3 series fatty acids, n-6 series fatty acids and total fatty acids (TFA) than diploid rainbow trout (P  0.05). Based on PUFA/SFA, UFA/SFA, PUFA/TFA, atherogenic index, and thrombogenic index, both diploid and triploid rainbow trout consumption were beneficial for human health, triploid fish had an even greater advantage. 3) Triploid rainbow trout had higher content of sodium, calcium, iron, manganese and zinc but lower content of magnesium, copper and selenium than diploid rainbow trout (P < 0.05). Multivariate statistical analysis of 79 quality indicators also showed that the fillet nutritional composition of triploid rainbow trout was different with its diploid counterparts. The reason may be related to the difference of lipid metabolism in rainbow trout with different ploidy

LINC00921 reduces lung cancer radiosensitivity by destabilizing NUDT21 and driving aberrant MED23 alternative polyadenylation

Summary: Alternative polyadenylation (APA) plays a major role in controlling transcriptome diversity and therapeutic resistance of cancers. However, long non-coding RNAs (lncRNAs) involved in pathological APA remain poorly defined. Here, we functionally characterize LINC00921, a MED13L/P300-induced oncogenic lncRNA, and show that it is required for global regulation of APA in non-small cell lung cancer (NSCLC). LINC00921 shows significant potential for reducing NSCLC radiosensitivity, and high LINC00921 levels are associated with a poor prognosis for patients with NSCLC treated with radiotherapy. LINC00921 controls NUDT21 stability by facilitating binding of NUDT21 with the E3 ligase TRIP12. LINC00921-induced destabilization of NUDT21 promotes 3′ UTR shortening of MED23 mRNA via APA, which, in turn, leads to elevated MED23 protein levels in cancer cells and nuclear translocation of β-catenin and thereby activates expression of multiple β-catenin/T cell factor (TCF)/lymphoid enhancer-binding factor (LEF)-regulated core oncogenes (c-Myc, CCND1, and BMP4). These findings highlight the importance of functionally annotating lncRNAs controlling APA and suggest the clinical potential of therapeutics for advanced NSCLC

Targeted exosome coating gene-chem nanocomplex as "nanoscavenger" for clearing alpha-synuclein and immune activation of Parkinson's disease

The most critical problem in the treatment of neurodegenerative diseases is brain neuronal protection, which can be overcome by clearing pathological substances and regulating the immune environment. In the above treatment strategies, the traditional poor drug delivery problem is inevitable. Here, we show an engineering core-shell hybrid system named rabies virus glycoprotein (RVG) peptide-modified exosome (EXO) curcumin/phenylboronic acid-poly(2-(dimethylamino)ethyl acrylate) nanoparticle/small interfering RNA targeting SNCA (REXO-C/ANP/S). It is a nanoscavenger for clearing alpha-synuclein aggregates and reducing their cytotoxicity in Parkinson's disease neurons. The motor behavior of Parkinson's disease mice is substantially improved after REXO-C/ANP/S treatment. In particular, we demonstrate that REXO-C/ANP/S is also a nanoscavenger for clearing immune activation due to its natural immature dendritic cell EXO coating. Our findings show that REXO-C/ANP/S may serve as a platform for neurodegenerative diseases treatment