Fig. S1The total N fertilizer input amount under different treatments.Different letters indicate significant differences at the level of P < 0.05.N+PK, straw return plus regular inorganic N-P-K fertilizers; 0.75N+PK, straw return plus regular inorganic P-K with 25% N fertilizer reduction; 0.5N+PK, straw return plus regular inorganic P-K with 50% N fertilizer reduction; PK, straw return plus regular inorganic P-K without N fertilizer.
<p>Pathway inhibitors Trametinib and Copanlisib inhibit the uptake and effect of EVs from various breast cancer cells. <b>A,</b> Western blots of lung fibroblasts treated with 200 nmol/L of Trametinib or Copanlisib or with 10 µg/mL EV for 24 hours. <b>B,</b> qRT-PCR–determined mRNA levels of S100A4 and FN1 with or without MDA-MB-231 EV treatment for 48 hours. Trametinib and Copanlisib were added together with EVs when indicated. Data were normalized to GAPDH. <b>C,</b> Lung fibroblasts were pretreated with Trametinib or Copanlisib (200 mmol/L) for 24 hours before Lck-GFP–labeled EVs derived from 4T1 or MDA-MB-468 cells were added and incubated for 6 hours. EV uptake was assessed and compared with DMSO-treated (No drug) control group. <b>D,</b> NIH3T3 fibroblasts were pretreated with Trametinib or Copanlisib before Lck-GFP–labeled EVs derived from 4T1 or MDA-MB-231 cells were added. Data are represented as mean ± SD (<i>n</i> = 3 wells). **, <i>P</i> < 0.01; ***, <i>P</i> < 0.001.</p>
Abstract. Nitrogen (N) fertilization has received worldwide attention due to its benefits to soil fertility and productivity, but excess N application also causes an array of ecosystem dis-services, such as greenhouse gas emissions. Generally, soil microorganisms are considered to be involved in upholding a variety of ecosystem services and dis-services. However, the linkages between soil ecosystem services and microbial traits under different N fertilizer application rates remain uncertain. To address this, a 4-year in situ field experiment was conducted in a meadow soil on the Northeast China Plain after straw return with the following treatments combined with regular phosphorus (P) and potassium (K) fertilization: (i) regular N fertilizer (N+PK); (ii) 25 % N fertilizer reduction (0.75N+PK); (iii) 50 % N fertilizer reduction (0.5N+PK); and (IV) no N fertilizer (PK). Ecosystem services, dis-services and microbial traits responded distinctly to the different N fertilizer rates. Treatment 0.75N+PK had overall positive effects on soil fertility, productivity, straw decomposition, and microbial abundance and function and alleviated greenhouse effects due to N deficiency. Meanwhile, 0.75N+PK upregulated aboveground biomass and soil C:N and thus increased the abundance of genes encoding cellulose-degrading enzymes, which may imply the potential ability of C and N turnover. In addition, most observed changes in ecosystem services and dis-services were strongly associated with microbial modules and keystone taxa. Specifically, the Lasiosphaeriaceae-driven module 1 community promoted straw degradation and C and N release, while the Terrimonas-driven module 3 community contributed to production improvement, which was conducive to soil multifunctionality. Therefore, our results suggest that straw return with 25 % chemical N fertilizer reduction is optimal for achieving ecosystem services. This study highlights the importance of abiotic and biotic factors in soil health and supports green agricultural development by optimizing N fertilizer rates in meadow soil after straw return.
We present vibrational sum-frequency generation (VSFG) microscopy as a new label-free chemical imaging technique for tumor identification. This method combines the chemical-bond selectivity of vibrational spectroscopy with coherent interference of second-order coherent nonlinear optics. Using a fast line-scanning VSFG microscope, we obtained hyperspectral VSFG images of collagen I from both lung tissues bearing metastatic tumors and in tumor-free ones, which reveal drastic different spectral signatures: tumor samples exhibit large NH stretch (NHS) and CH stretch (CHS) versus the CH2 symmetric stretch (CH2,Ss), compared to healthy controls. We then identified two key spectral signatures to distinguish metastatic tumor and tumor-free tissues: the intensity ratio of NHS/CH2,Ss and CHS/CH2,Ss modes. These signatures demonstrated high fidelity in distinguishing between tumors and normal tissue in both mouse and human lung samples. Theoretical modeling indicates that distinctive interferences of spectral peaks are sensitive to interfibrillar distances, at 130 of nanometer level. These findings suggest that collagen fibrils are more densely packed in tumors, corroborating the enhanced stiffness observed in tumor tissues. VSFG microscopy offers a highly selective, label-free chemical imaging characterization that preserves sample integrity, making it a valuable tool for oncology, pathology and fundamental biophysical research.
<div>Abstract<p>Cancer-secreted, extracellular vesicle (EV)–encapsulated miRNAs enable cancer cells to communicate with each other and with noncancerous cells in tumor pathogenesis and response to therapies. Here, we show that treatment with a sublethal dose of chemotherapeutic agents induces breast cancer cells to secrete EV with the capacity to stimulate a cancer stem-like cell (CSC) phenotype, rendering cancer cells resistance to therapy. Chemotherapy induced breast cancer cells to secrete multiple EV miRNAs, including miR-9-5p, miR-195-5p, and miR-203a-3p, which simultaneously targeted the transcription factor One Cut Homeobox 2 (ONECUT2), leading to induction of CSC traits and expression of stemness-associated genes, including <i>NOTCH1, SOX9, NANOG, OCT4</i>, and <i>SOX2</i>. Inhibition of these miRNAs or restoration of ONECUT2 expression abolished the CSC-stimulating effect of EV from chemotherapy-treated cancer cells. In mice bearing xenograft mammary tumors, docetaxel treatment caused elevations of miR-9-5p, miR-195-5p, and miR-203a-3p in circulating EV and decreased ONECUT2 expression and increased levels of stemness-associated genes. These effects following chemotherapy were diminished in tumors deficient in exosome secretion. In human breast tumors, neoadjuvant chemotherapy decreased ONECUT2 expression in tumor cells. Our results indicate a mechanism by which cancer cells communicate with each other and self-adapt to survive in response to cytotoxic treatment. Targeting these adaptation mechanisms along with chemotherapy, such as by blocking the EV miRNA–ONECUT2 axis, represents a potential strategy to maximize the anticancer effect of chemotherapy and to reduce chemoresistance in cancer management.</p>Significance:<p>These findings reveal a critical mechanism of resistance to chemotherapy by which breast cancer cells secrete miRNA-containing extracellular vesicles to stimulate cancer stem cell-like features.</p></div>
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