Intramyocardial Injection of Recombinant Adeno-Associated Viral Vector Coexpressing PR39/Adrenomedullin Enhances Angiogenesis and Reduces Apoptosis in a Rat Myocardial Infarction Model

Cotransfer of angiogenic and antiapoptotic genes could be the basis of new gene therapy strategies for myocardial infarction. In this study, rAAV-PR39-ADM, coexpressing antimicrobial peptide (PR39) and adrenomedullin (ADM), was designed with the mediation of recombinant adeno-associated virus. In vitro, CRL-1730 cells were divided into four groups, namely, the sham group, the AAV-null group, the NS (normal saline) group, and the PR39-ADM group. Immunocytochemistry analysis, CCK-8 assays, Matrigel assays, and apoptotic analysis were performed; in vivo, myocardial infarction model was established through ligation of the left coronary artery on rats, and treatment groups corresponded to those used in vitro. Myocardial injury, cardiac performance, and the extent of myocardial apoptosis were assessed. Results suggested that rAAV-PR39-ADM administration after myocardial infarction improved cell viability and cardiac function, attenuated apoptosis and myocardial injury, and promoted angiogenesis. Subsequently, levels of 6×His, HIF-1α, VEGF, p-Akt, Akt, ADM, Bcl-2, and Bax were measured by western blot. rAAV-PR39-ADM increased p-Akt, HIF-1α, and VEGF levels and induced higher Bcl-2 expression and lower Bax expression. In conclusion, our results demonstrate that rAAV-PR39-ADM mitigates myocardial injury by promoting angiogenesis and reducing apoptosis. This study suggests a potential novel gene therapy-based method that could be used clinically for myocardial infarction

Control of Streptomyces alfalfae XY25T Over Clubroot Disease and Its Effect on Rhizosphere Microbial Community in Chinese Cabbage Field Trials

Clubroot caused by Plasmodiophora brassicae is one of the most destructive diseases in cruciferous crops. Streptomyces alfalfae XY25T, a biological control agent, exhibited great ability to relieve clubroot disease, regulate rhizosphere bacterial and fungal communities in Chinese cabbage, and promote its growth in greenhouse. Therefore, field experiments were carried out to investigate the effects of S. alfalfae XY25T on clubroot and rhizosphere microbial community in Chinese cabbage. Results showed that the control efficiency of clubroot by S. alfalfae XY25T was 69.4%. Applying the agent can alleviate soil acidification; increase the contents of soil organic matter, available nitrogen, available phosphorus, and available potassium; and enhance activities of invertase, urease, catalase, and alkaline phosphatase. During Chinese cabbage growth, bacterial diversity decreased first and then increased, and fungal diversity decreased gradually after inoculation with S. alfalfae XY25T. High-throughput sequencing analysis showed that the main bacterial phyla were Proteobacteria, Bacteroidetes, Acidobacteria, and Planctomycetes, and the major fungal phyla were Ascomycota and Basidiomycota in rhizosphere soil. The dominant bacterial genera were Flavobacterium, Candidatus, Pseudomonas, Stenotrophomonas, Sphingomonas, Flavisolibacter, and Gemmatimonbacteria with no significant difference in abundance, and the major fungal genera were Monographella, Aspergillus, Hypocreales, Chytridiaceae, Fusarium, Pleosporales, Agaricales, Mortierella, and Pleosporales. The significant differences were observed among Pleosporales, Basidiomycota, Colletotrichum, two strains attributed to Agaricales, and another two unidentified fungi by using S. alfalfae XY25T. Moreover, quantitative real-time PCR results indicated that P. brassicae content was significantly decreased after the agent inoculation. In conclusion, S. alfalfae XY25T can affect rhizosphere microbial communities; therefore, applying the agent is an effective approach to reduce the damage caused by clubroot

Identification and Verification of Candidate miRNA Biomarkers with Application to Infection with <i>Emiliania huxleyi</i> Virus

The interactions of Emiliania huxleyi and its specific lytic virus (EhV) have a profound influence on marine biogeochemical carbon–sulfur cycles and play a prominent role in global climate change. MicroRNAs (miRNAs) have emerged as promising candidates with extensive diagnostic potential due to their role in virus–host interactions. However, the application of miRNA signatures as diagnostic markers in marine viral infection has made limited progress. Based on our previous small-RNA sequencing data, one host miRNA biomarker that is upregulated in early infection and seven viral miRNA biomarkers that are upregulated in late infection were identified and verified using qRT-PCR and a receiver operating characteristic curve analysis in pure culture, mixed culture, and natural seawater culture. The host ehx-miR20-5p was able to significantly differentiate infection groups from the control in the middle (24 h post-infection, hpi) and late infection (48 hpi) phases, while seven virus-derived miRNA biomarkers could diagnose the early and late stages of EhV infection. Functional enrichment analysis showed that these miRNAs participated in numerous essential metabolic pathways, including gene transcription and translation, cell division-related pathways, protein-degradation-related processes, and lipid metabolism. Additionally, a dual-luciferase reporter assay confirmed the targeted relationship between a viral ehv-miR7-5p and the host dihydroceramide desaturase gene (hDCD). This finding suggests that the virus-derived miRNA has the ability to inhibit the host sphingolipid metabolism, which is a specific characteristic of EhV infection during the late stage. Our data revealed a cluster of potential miRNA biomarkers with significant regulatory functions that could be used to diagnose EhV infection, which has implications for assessing the infectious activity of EhV in a natural marine environment

Advances and Limitations of Next Generation Sequencing in Animal Diet Analysis

Diet analysis is a critical content of animal ecology and the diet analysis methods have been constantly improving and updating. Contrary to traditional methods of high labor intensity and low resolution, the next generation sequencing (NGS) approach has been suggested as a promising tool for dietary studies, which greatly improves the efficiency and broadens the application range. Here we present a framework of adopting NGS and DNA metabarcoding into diet analysis, and discuss the application in aspects of prey taxa composition and structure, intra-specific and inter-specific trophic links, and the effects of animal feeding on environmental changes. Yet, the generation of NGS-based diet data and subsequent analyses and interpretations are still challenging with several factors, making it possible still not as widely used as might be expected. We suggest that NGS-based diet methods must be furthered, analytical pipelines should be developed. More application perspectives, including nutrient geometry, metagenomics and nutrigenomics, need to be incorporated to encourage more ecologists to infer novel insights on they work

[(η5-pentamethylcyclopentadienyl)(3-fluoro-N-methylbenzylamine-к1,N)dichlorido]iridium(III)

A half-sandwich iridium(III) complex containing 3-fluoro-N-methylbenzylamine ligands has been obtained by reaction of one equivalent of [(&eta;5-Cp*)IrCl2]2 (Cp* = pentamethylcyclopentadienyl) with two equivalent of 3-fluoro-N-methylbenzylamine in very good yield. The structure of this complex was confirmed by X-ray crystallography, 1H-NMR, 13C-NMR spectroscopy, and elemental analysis

2,4-Bis[(2,6-diisopropylphenyl)imino]-3-methylpentan-3-ol

The compound 2,4-bis[(2,6-diisopropylphenyl)imino]-3-methylpentan-3-ol was synthesized with a yield of approximately 80% by the reaction of 2,4-bis(2,6-diisopropylphenylimino)pentan-3-one with trimethylaluminum, which was followed by hydrolysis with an aqueous NaOH solution. A chemoselective addition to the C=O bond occurred in this reaction. The structure was confirmed by X-ray crystallography. This new compound was also fully characterized by 1H, 13C-NMR spectroscopy and 1H-13C HSQC spectroscopy, mass spectrometry and elemental analysis

Synthesis and Reactivity of N‑Heterocyclic PSiP Pincer Iron and Cobalt Complexes and Catalytic Application of Cobalt Hydride in Kumada Coupling Reactions

The new N-heterocyclic σ-silyl pincer ligand HSiMe­(NCH₂PPh₂)₂C₆H₄ (<b>1</b>) was designed. A series of tridentate silyl pincer Fe and Co complexes were prepared. Most of them were formed by chelate-assisted Si–H activation. The typical iron hydrido complex FeH­(PMe₃)₂(SiMe­(NCH₂PPh₂)₂C₆H₄) (<b>2</b>) was obtained by Si–H activation of compound <b>1</b> with Fe­(PMe₃)₄. The combination of compound <b>1</b> with CoMe­(PMe₃)₄ afforded the Co­(I) complex Co­(PMe₃)₂(SiMe­(NCH₂PPh₂)₂C₆H₄) (<b>3</b>). The Co­(III) complex CoHCl­(PMe₃)­(SiMe­(NCH₂PPh₂)₂C₆H₄) (<b>5</b>) was generated by the reaction of complex <b>1</b> with CoCl­(PMe₃)₃ or the combination of complex <b>3</b> with HCl. However, when complex <b>3</b> was treated with MeI, the Co­(II) complex CoI­(PMe₃)­(SiMe­(NCH₂PPh₂)₂C₆H₄) (<b>4</b>), rather than the Co­(III) complex, was isolated. The catalytic performance of complex <b>5</b> for Kumada coupling reactions was explored. With a catalyst loading of 5 mol %, complex <b>5</b> displayed efficient catalytic activity for Kumada cross-coupling reactions of aryl chlorides and aryl bromides with Grignard reagents. This catalytic reaction mechanism is proposed and partially experimentally verified

The application effect of percutaneous cryoablation for the stage IIIB/IV advanced non-small-cell lung cancer after the failure of chemoradiotherapy

Summary: Background/Objective: A retrospective review is required to assess the application effect of percutaneous cryoablation on the stage IIIB/IV advanced non-small-cell lung cancer (NSCLC) after the failure of chemoradiotherapy. A total of twenty-two cases with stage IIIB/IV advanced NSCLC after failure of chemoradiotherapy were enrolled. Methods: Computed Tomography (CT) was used for the assessment of tumor response, and Functional Assessment of Cancer Therapy-General (FACT-G) was applied for the evaluation of quality of life. After the treatment of cryoablation, we performed a follow-up analysis. Results: The primary technique effectiveness of 100% was observed in the one month of follow-up. At three months, local tumor progression was shown in 4 of 31 lesions (13.6%). The one-year survival rate of 81.8% and progression-free rate of 27.8% were obtained. Six patients died 7, 9, 10, 12, 15 and 22 months after treatment, respectively. Decreased scores of total quality of life at one week after cryoablation (P = 0.006), but increased scores at one month after cryoablation (P = 0.024) were observed, compared with pre-cryoablation. Conclusion: Cryoablation contributes to the effective local tumor therapy for stage IIIB/IV advanced NSCLC after the failure of chemoradiotherapy. Keywords: Cryoablation, Lung cancer, Stage IIIB/I

Surface Morphology and Microstructure Evolution of Single Crystal Diamond during Different Homoepitaxial Growth Stages

Homoepitaxial growth of step-flow single crystal diamond was performed by microwave plasma chemical vapor deposition system on high-pressure high-temperature diamond substrate. A coarse surface morphology with isolated particles was firstly deposited on diamond substrate as an interlayer under hillock growth model. Then, the growth model was changed to step-flow growth model for growing step-flow single crystal diamond layer on this hillock interlayer. Furthermore, the surface morphology evolution, cross-section and surface microstructure, and crystal quality of grown diamond were evaluated by scanning electron microscopy, high-resolution transmission electron microcopy, and Raman and photoluminescence spectroscopy. It was found that the surface morphology varied with deposition time under step-flow growth parameters. The cross-section topography exhibited obvious inhomogeneity in crystal structure. Additionally, the diamond growth mechanism from the microscopic point of view was revealed to illustrate the morphological and structural evolution