Effect of nitrogen addition on soil net nitrogen mineralization in topsoil and subsoil regulated by soil microbial properties and mineral protection: Evidence from a long-term grassland experiment.

Soil net nitrogen mineralization (Nmin), a microbial-mediated conversion of organic to inorganic N, is critical for grassland productivity and biogeochemical cycling. Enhanced atmospheric N deposition has been shown to substantially increase both plant and soil N content, leading to a major change in Nmin. However, the mechanisms underlying microbial properties, particularly microbial functional genes, which drive the response of Nmin to elevated N deposition are still being discussed. Besides, it is still uncertain whether the relative importance of plant carbon (C) input, microbial properties, and mineral protection in regulating Nmin under continuous N addition would vary with the soil depth. Here, based on a 13-year multi-level field N addition experiment conducted in a typical grassland on the Loess Plateau, we elucidated how N-induced changes in plant C input, soil physicochemical properties, mineral properties, soil microbial community, and the soil Nmin rate (Rmin)-related functional genes drove the responses of Rmin to N addition in the topsoil and subsoil. The results showed that Rmin increased significantly in both topsoil and subsoil with increasing rates of N addition. Such a response was mainly dominated by the rate of soil nitrification. Structural equation modeling (SEM) revealed that a combination of microbial properties (functional genes and diversity) and mineral properties regulated the response of Rmin to N addition at both soil depths, thus leading to changes in the soil N availability. More importantly, the regulatory impacts of microbial and mineral properties on Rmin were depth-dependent: the influences of microbial properties weakened with soil depth, whereas the effects of mineral protection enhanced with soil depth. Collectively, these results highlight the need to incorporate the effects of differential microbial and mineral properties on Rmin at different soil depths into the Earth system models to better predict soil N cycling under further scenarios of N deposition.

S-allylmercapto-N-acetylcysteine ameliorates pulmonary fibrosis in mice via Nrf2 pathway activation and NF-κB, TGF-ß1/Smad2/3 pathway suppression.

Pulmonary fibrosis (PF) is a chronic lung disease characterised by alveolar inflammatory injury, alveolar septal thickening, and eventually fibrosis. Patients with severe Coronavirus Disease 2019 (COVID-19) may have left a certain degree of pulmonary fibrosis. PF is commonly caused by oxidative imbalance and inflammatory damage. S-allylmercapto-N-acetylcysteine (ASSNAC) exhibits anti-oxidative and anti-inflammatory effects in other diseases. However, the pharmacodynamics of ASSNAC remain unclear for PF. This investigation aimed to evaluate the efficacy and mechanism of ASSNAC against PF. The PF model was established by TGF-ß1 stimulating HFL-1 cells in vitro. ASSNAC exhibited the potential to inhibit fibroblast transformation into myofibroblasts. Also, in the PF mice model with bleomycin (BLM), the sodium salt of ASSNAC (ASSNAC-Na) inhalation was treated. ASSNAC remarkably improved mice's lung tissue structure and collagen deposition. The important indicator proteins of PF, collagen Ⅰ, collagen Ⅲ, and α-SMA significantly decreased in the ASSNAC treated groups. Besides, ASSNAC attenuated oxidative stress by reversing glutathione (GSH), superoxide dismutase (SOD) levels and interfering with Nrf2/NOX4 signaling pathways. ASSNAC showed an anti-inflammatory effect by reducing the number of inflammatory cells and inflammatory cytokines, such as TNF-α and IL-6, and blocking the NF-κB signaling pathway. ASSNAC inhibited fibroblast differentiation by blocking the TGF-ß1/Smad2/3 signaling pathway. This study implicates that ASSNAC alleviates pulmonary fibrosis through fighting against oxidative stress, reducing inflammation and inhibiting fibroblast differentiation.

The epiphany derived from T-cell-inflamed profiles: Pan-cancer characterization of CD8A as a biomarker spanning clinical relevance, cancer prognosis, immunosuppressive environment, and treatment responses.

CD8A encodes the CD8 alpha chain of αßT cells, which has been proposed as a quantifiable indicator for the assessment of CD8+ cytotoxic T lymphocytes (CTLs) recruitment or activity and a robust biomarker for anti-PD-1/PD-L1 therapy responses. Nonetheless, the lack of research into the role of CD8A in tumor microenvironment predisposes to limitations in its clinical utilization. In the presented study, multiple computational tools were used to investigate the roles of CD8A in the pan-cancer study, revealing its essential associations with tumor immune infiltration, immunosuppressive environment formation, cancer progression, and therapy responses. Based on the pan-cancer cohorts of the Cancer Genome Atlas (TCGA) database, our results demonstrated the distinctive CD8A expression patterns in cancer tissues and its close associations with the prognosis and disease stage of cancer. We then found that CD8A was correlated with six major immune cell types, and immunosuppressive cells in multiple cancer types. Besides, epigenetic modifications of CD8A were related to CTL levels and T cell dysfunctional states, thereby affecting survival outcomes of specific cancer types. After that, we explored the co-occurrence patterns of CD8A mutation, thus identifying RMND5A, RNF103-CHMP3, CHMP3, CD8B, MRPL35, MAT2A, RGPD1, RGPD2, REEP1, and ANAPC1P1 genes, which co-occurred mutations with CD8A, and are concomitantly implicated in the regulation of cancer-related pathways. Finally, we tested CD8A as a therapeutic biomarker for multiple antitumor agents' or compounds' responsiveness on various cancer cell lines and cancer cohorts. Our findings denoted the underlying mechanics of CD8A in reflecting the T-cell-inflamed profiles, which has potential as a biomarker in cancer diagnosis, prognosis, and therapeutic responses.

Urine Exosomal AMACR Is a Novel Biomarker for Prostate Cancer Detection at Initial Biopsy.

Objectives: The aim of this study is to identify and validate urine exosomal AMACR (UE-A) as a novel biomarker to improve the detection of prostate cancer (PCa) and clinically significant PCa (Gleason score ≥ 7) at initial prostate biopsy. Methods: A total of 289 first-catch urine samples after the digital rectal exam (DRE) were collected from patients who underwent prostatic biopsy, and 17 patients were excluded due to incomplete clinical information. Urine exosomes were purified, and urinary exosomal AMACR (UE-A) was measured by enzyme-linked immunosorbent assay (ELISA). The diagnostic performance of UE-A was evaluated by receiver operating characteristic (ROC) analysis, decision curve analysis (DCA), and waterfall plots. Results: The expression of AMACR in PCa and csPCa was significantly higher than that in BPH and non-aggressive (p < 0.001). The UE-A presented good performance in distinguishing PCa from BPH or BPH plus non-significant PCa (nsPCa) from csPCa with an area under the ROC curve (AUC) of 0.832 and 0.78, respectively. The performance of UE-A was further validated in a multi-center cohort of patients with an AUC of 0.800 for detecting PCa and 0.749 for detecting csPCa. The clinical utility assessed by DCA showed that the benefit of patients using UE-A was superior to PSA, f/t PSA, and PSAD in both the training cohort and the validation cohort in terms of all threshold probabilities. Setting 95% sensitivity as the cutoff value, UE-A could avoid 27.57% of unnecessary biopsies, with only 4 (1.47%) csPCa patients missed. Conclusions: We demonstrated the great performance of UE-A for the early diagnosis of PCa and csPCa. UE-A could be a novel non-invasive diagnostic biomarker to improve the detection of PCa and csPCa.

Transcriptome Analysis of Zygophyllum xanthoxylum Adaptation Strategies to Phosphate Stress.

Soil phosphate (Pi) deficiency is a global issue and a major constraint on plant growth. Plants typically acclimatize to low Pi by enhancing their P utilization and/or P acquisition efficiencies; however, different species have variable preferred strategies. RNA sequencing analysis was performed on the shoots and roots of Zygophyllum xanthoxylum, under 1 day and 10 days of Pi stress, to investigate their adaptation strategies to P deprivation. A total of 364,614 unigenes and 9,270 differentially expressed genes (DEGs) were obtained via transcriptome sequencing. An analysis of the DEGs revealed that under the 10D treatment, anthocyanin synthesis genes were upregulated under Pi stress, whereas gibberellin, ethylene, and cytokinins synthesis genes were upregulated, and abscisic acid synthesis genes were downregulated. Genes related to organic acid synthesis, encoding for purple acid phosphatases (APase) and nucleases (RNase) were upregulated under the 1D and 10D treatments, respectively. Furthermore, genes associated with Pi transport were induced by Pi stress. Zygophyllum xanthoxylum has special P adaptation strategies, the variation trends of genes involved in external P mobilization and acquisition, which were different from that of most other species; however, the expression levels of organophosphorus mobilization related genes, such as APases and RNases, were significantly increased. Meanwhile, PHT2s and TPTs, which distributed Pi to effective sites (e.g., chloroplast), played critical roles in the maintenance of photosynthesis. We speculated that these were economic and energy saving strategies, and there are critical adaptive mechanisms that Z. xanthoxylum employs to cope with deficits in Pi.

Transcriptome and co-expression network analysis reveal molecular mechanisms of mucilage formation during seed development in Artemisia sphaerocephala.

Seed mucilage has significant economic value. However, the identification of key regulatory genes in mucilage formation and their molecular regulatory mechanism remain unknown. Artemisia sphaerocephala seeds are rich in mucilage. In this study, A. sphaerocephala seeds in 10, 20, 30, 40, 50, 60 and 70 days after flowering were used as materials to reveal their molecular regulatory mechanism in mucilage formation by RNA-sequencing and weighted gene co-expression network analysis (WGCNA). 21 key regulatory genes for mucilage formation were identified, including AsKNAT7 and AsTTG1 genes, as well as AsNAM and AsAP2 gene families. From 10-30 days after flowering, both AsNAM and AsAP2 supported mucilage formation. From 40-70 days after flowering, promotion by AsNAM and AsAP2 was weakened and the up-regulation of AsKNAT7 inhibited mucilage formation, leading to no further increases in mucilage content. This in depth elucidation of seed mucilage formation lays the foundation for the application of mucilage.

Bacterial Communities in Stream Biofilms in a Degrading Grassland Watershed on the Qinghai-Tibet Plateau.

Grassland is among the largest terrestrial biomes and is experiencing serious degradation, especially on the Qinghai-Tibet Plateau (QTP). However, the influences of grassland degradation on microbial communities in stream biofilms are largely unknown. Using 16S rRNA gene sequencing, we investigated the bacterial communities in stream biofilms in sub-basins with different grassland status in the Qinghai Lake watershed. Grassland status in the sub-basins was quantified using the normalized difference vegetation index (NDVI). Proteobacteria, Bacteroidetes, Cyanobacteria, and Verrucomicrobia were the dominant bacterial phyla. OTUs, 7,050, were detected in total, within which 19 were abundant taxa, and 6,922 were rare taxa. Chao 1, the number of observed OTUs, and phylogenetic diversity had positive correlations with carbon (C), nitrogen (N), and/or phosphorus (P) in biofilms per se. The variation of bacterial communities in stream biofilms was closely associated with the rate of change in NDVI, pH, conductivity, as well as C, N, P, contents and C:N ratio of the biofilms. Abundant subcommunities were more influenced by environmental variables relative to the whole community and to rare subcommunities. These results suggest that the history of grassland degradation (indicated as the rate of change in NDVI) influences bacterial communities in stream biofilms. Moreover, the bacterial community network showed high modularity with five major modules (>50 nodes) that responded differently to environmental variables. According to the module structure, only one module connector and 12 module hubs were identified, suggesting high fragmentation of the network and considerable independence of the modules. Most of the keystone taxa were rare taxa, consistent with fragmentation of the network and with adverse consequences for bacterial community integrity and function in the biofilms. By documenting the properties of bacterial communities in stream biofilms in a degrading grassland watershed, our study adds to our knowledge of the potential influences of grassland degradation on aquatic ecosystems.

Effects of grassland degradation on ecological stoichiometry of soil ecosystems on the Qinghai-Tibet Plateau.

Grasslands across the world are being degraded due to the impacts of overgrazing and climate change. However, the influences of grassland degradation on carbon (C), nitrogen (N), and phosphorus (P) dynamics and stoichiometry in soil ecosystems are not well studied, especially at high elevations where ongoing climate change is most pronounced. Ecological stoichiometry facilitates understanding the biogeochemical cycles of multiple elements by studying their balance in ecological systems. This study sought to assess the responses of these soil elements to grassland degradation in the Qinghai Lake watershed on the Qinghai-Tibet Plateau (QTP), which has an average elevation of >4000 m and is experiencing serious grassland degradation due to its sensitivity and vulnerability to external disturbances. Substituting space for time, we quantified normalized difference vegetation index to gauge grassland degradation. C, N, and P concentrations and their molar ratios in soil and in soil microbial biomass were also measured. The results showed that grassland degradation decreased the concentrations of C and N, as well as the ratios of C:P and N:P in soil. The soil became relatively more P rich and thus N limitation is anticipated to be more apparent with grassland degradation. Moreover, C, N, and P concentrations in soil microbial biomass decreased with increased grassland degradation. C:N:P ratios of soil microbial biomass were highly constrained, suggesting that soil microorganisms exhibited a strong homeostatic behavior, while the variations of microbial biomass C:N:P ratios suggest changes in microbial activities and community structure. Overall, our study revealed that grassland degradation differentially affects soil C, N, and P, leading to decreased C:N and N:P in soil, as well as decreased C, N, and P concentrations in soil microbial biomass. This study provides insights from a stoichiometric perspective into microbial and biogeochemical responses of grassland ecosystems as they undergo degradation on the QTP.

Cascading influences of grassland degradation on nutrient limitation in a high mountain lake and its inflow streams.

Nitrogen (N) and phosphorus (P) are key growth-limiting nutrients for organisms and their absolute and relative supplies regulate the structure and function of ecosystems. Landcover changes lead to modifications of terrestrial biogeochemistry, consequently influencing aquatic nutrient conditions. This study sought to evaluate the potential impacts of grassland degradation on nutrient availability and nutrient limitation in the Qinghai Lake (China) and its inflow streams. We sampled nutrient concentrations and tested stream nutrient limitation by conducting nutrient diffusing substrata (NDS) bioassays in streams flowing through subbasins with different grassland status. To test nutrient limitation and the responses of lake phytoplankton to stream inflows, bioassays were conducted by adding different nutrients (N, P, and joint NP) as well as water from different streams to lake water with phytoplankton, respectively. In general, N concentrations as well as N:P ratios decreased while P concentrations increased with decreased normalized difference vegetation index (NDVI, an index of vegetation status), especially in September, suggesting that grassland degradation (low NDVI) has the potential to differentially decrease N availability and increase P availability in streams. Consistent with this, relative responses (RR) of stream periphyton to P and combined NP enrichments in the NDS bioassays decreased with stream P concentrations while increased with stream water N:P ratios. Lake phytoplankton responded strongly to P and combined NP addition indicating strong P-limitation of lake phytoplankton. RR of lake phytoplankton to stream water decreased with nitrate concentration and N:P ratios in stream water and increased with the concentrations of ammonium, total phosphorus, and soluble reactive phosphorus, indicating that stream water with higher P but lower N and N:P from degraded subcatchments is associated with increased impact on P-limited Lake phytoplankton. Overall, this study suggests that grassland degradation has the potential to differentially influence the nutrients delivered to streams with substantial increases in P but decreases in N and N:P, alleviating P limitation of stream periphyton and, ultimately, stimulating P-limited phytoplankton growth in the lake.

The impact of nitrogen enrichment on grassland ecosystem stability depends on nitrogen addition level.

Increasing atmospheric nitrogen (N) deposition may affect plant biodiversity, subsequently altering ecosystem stability. While a few studies have explored how simulated N deposition affects community stability and its underlying mechanisms, the experimental levels of N addition used are usually higher than current and future N deposition rates. Thus, their results could produce highly uncertain predictions of ecosystem function, especially if the responses to N deposition are nonlinear. We conducted a manipulative experiment that simulated elevated atmospheric N deposition with several N addition levels to evaluate the effect of N deposition on ecosystem stability and its underlying mechanisms in a semiarid grassland in northern China. Here we show that N addition altered community diversity, reducing species richness, evenness, diversity and dominance. In addition, we found that N addition at current N deposition levels had no significant impact on community stability. In contrast, N addition at levels from 4.6 to 13.8gNm-2yr-1 significantly decreased community stability, although community stability for the 13.8gNm-2yr-1 treatment was higher than that for the 4.6gNm-2yr-1 treatment. These results indicate that the response of community stability to N enrichment is nonlinear. This nonlinear change in community stability was positively correlated with species asynchrony, species richness, and species diversity as well as the stability of dominant species and the stability of the grass functional group. Our data suggest a need to re-evaluate the mechanisms responsible for the effects of N deposition on natural ecosystem stability across multiple levels of N enrichment and that additional experimentation with gradients of N loads more similar to future atmospheric N deposition rates is needed.

Contribution of root respiration to total soil respiration in a semi-arid grassland on the Loess Plateau, China.

Using the trenching method, a study was conducted in a grassland on the Loess Plateau of northern China in 2008 and 2009 to partition total soil respiration (Rt) into microbial respiration (Rm) and root respiration (Rr). Using the measurements of soil CO2 diffusivity and soil CO2 production, an analytical model was applied to correct the data, aiming to quantify the method-induced error. The results showed that Rm and Rr responded differently to biotic and abiotic factors and exhibited different diurnal and seasonal variations. The diurnal variation of Rm was strongly controlled by soil temperature, while Rr might be mainly controlled by photosynthesis. The combination of soil temperature and moisture could better explain the seasonal variation in Rm (r2=0.76, P<0.001). The seasonal variation of Rr was influenced mainly by the plant activity. The contribution of root respiration to total soil respiration (Rr/Rt ratio) also exhibited substantial diurnal and seasonal variations, being higher at nighttime and lower at daytime. In the different growing stages, the Rr/Rt ratios ranged from 15.0% to 62.0% in 2008 and 14.5% to 63.6% in 2009. The mean values of the Rr/Rt ratio in the growing season and the annual mean Rr/Rt ratio were 41.7% and 41.9%, respectively, during the experiment period. Different precipitation distributions in the two years did not change the yearly Rr/Rt ratio. Corrected with the analytical model, the trenching method in small root-free plots led to an underestimation of Rr and Rr/Rt ratio by 4.2% and 1.8%.