Biparatopic anti-PCSK9 antibody enhances the LDL-uptake in HepG2 cells.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) has emerged as a promising therapeutic target to reduce lipids. In 2020, we reported a chimeric camelid-human heavy chain antibody VHH-B11-Fc targeting PCSK9. Recently, it was verified that VHH-B11 binds one linear epitope in the PCSK9 hinge region. To enhance its druggability, we have developed a novel biparatopic B11-H2-Fc Ab herein. Thereinto, surface plasmon resonance (SPR) confirmed the epitope differences in binding-PCSK9 among VHH-B11, VHH-H2 and the approved Repatha. Additionally, SPR revealed the B11-H2-Fc exhibits an avidity of approximately 0.036 nM for PCSK9, representing a considerable increase compared to VHH-B11-Fc (~ 0.69 nM). Moreover, we found the Repatha and B11-H2-Fc exhibited > 95% PCSK9 inhibition efficiency compared to approximately 48% for the VHH-Fc at 7.4 nM (P < 0.0005). Further, we verified its biological activity using the human hepatoma cells G2 model, where the B11-H2-Fc exhibited almost 100% efficiency in PCSK9 inhibition at only 0.75 µM. The immunoblotting results of low-density lipoprotein cholesterol (LDL-c) uptake assay also demonstrated the excellent performance of B11-H2-Fc on recovering the LDL-c receptor (LDLR), as strong as the Repatha (P > 0.05). These findings provide the first evidence of the efficacy of a novel Ab targeting PCSK9 in the field of lipid-lowering drugs.

A national-scale assessment of land subsidence in China's major cities.

China's massive wave of urbanization may be threatened by land subsidence. Using a spaceborne synthetic aperture radar interferometry technique, we provided a systematic assessment of land subsidence in all of China's major cities from 2015 to 2022. Of the examined urban lands, 45% are subsiding faster than 3 millimeters per year, and 16% are subsiding faster than 10 millimeters per year, affecting 29 and 7% of the urban population, respectively. The subsidence appears to be associated with a range of factors such as groundwater withdrawal and the weight of buildings. By 2120, 22 to 26% of China's coastal lands will have a relative elevation lower than sea level, hosting 9 to 11% of the coastal population, because of the combined effect of city subsidence and sea-level rise. Our results underscore the necessity of enhancing protective measures to mitigate potential damages from subsidence.

Does Forest Soil Fungal Community Respond to Short-Term Simulated Nitrogen Deposition in Different Forests in Eastern China?

Nitrogen (N) deposition has changed plants and soil microbes remarkably, which deeply alters the structures and functions of terrestrial ecosystems. However, how forest fungal diversity, community compositions, and their potential functions respond to N deposition is still lacking in exploration at a large scale. In this study, we conducted a short-term (4-5 years) experiment of artificial N addition to simulated N deposition in five typical forest ecosystems across eastern China, which includes tropical montane rainforest, subtropical evergreen broadleaved forest, temperate deciduous broadleaved forest, temperate broadleaved and conifer mixed forest, and boreal forest along a latitudinal gradient from tropical to cold temperature zones. Fungal compositions were identified using high-throughput sequencing at the topsoil layer. The results showed that fungal diversity and fungal community compositions among forests varied apparently for both unfertilized and fertilized soils. Generally, soil fungal diversity, communities, and their potential functions responded sluggishly to short-term N addition, whereas the fungal Shannon index was increased in the tropical forest. In addition, environmental heterogeneity explained most of the variation among fungal communities along the latitudinal gradient. Specifically, soil C: N ratio and soil water content were the most important factors driving fungal diversity, whereas mean annual temperature and microbial nutrient limitation mainly shaped fungal community structure and functional compositions. Topsoil fungal communities in eastern forest ecosystems in China were more sensitive to environmental heterogeneity rather than short-term N addition. Our study further emphasized the importance of simultaneously evaluating soil fungal communities in different forest types in response to atmospheric N deposition.

Different responses of soil bacterial and fungal communities to nitrogen deposition in a subtropical forest.

China has experienced a widespread increase in N deposition due to intensive anthropogenic activities, particularly in the subtropical regions. However, the effects of long-term N deposition on soil bacterial and fungal abundance, diversity, and community composition remain largely unclear. We assessed the effects of N deposition on soil microbial communities in summer and winter, using quantitative polymerase chain reaction and Illumina Miseq sequencing of bacterial 16S rRNA and fungal ITS genes from subtropical natural forest soils. The abundance of both soil bacteria and fungi exhibited a decreasing pattern with increasing N deposition rates. Nitrogen deposition increased bacterial diversity in both summer and winter, whereas fungal diversity was significantly decreased in summer, but greatly increased under the highest level of N deposition (150 kg N ha-1 yr-1) in winter. Nitrogen deposition significantly increased the relative abundance of bacterial phyla Actinobacteria, Chloroflexi, and WPS-2, but decreased that of Acidobacteria and Verrucomicrobia. In addition, N deposition significantly decreased the relative abundance of Ascomycetes, but did not exert any significant effect on Basidiomycetes. The bacterial and fungal community compositions were greatly influenced by N deposition, with soil N availability and soil pH identified as the two most influential soil properties. This study demonstrates that the fungal community was more sensitive than the bacterial community to N deposition, and further emphasizes the importance of simultaneously evaluating soil bacterial and fungal communities in response to global environmental changes.

Effects of nitrogen addition on microbial residues and their contribution to soil organic carbon in China's forests from tropical to boreal zone.

Atmospheric nitrogen (N) deposition has a significant influence on soil organic carbon (SOC) accumulation in forest ecosystems. Microbial residues, as by-products of microbial anabolism, account for a significant fraction of soil C pools. However, how N deposition affects the accumulation of soil microbial residues in different forest biomes remains unclear. Here, we investigated the effects of six/seven-year N additions on microbial residues (amino sugar biomarkers) in eight forests from tropical to boreal zone in eastern China. Our results showed a minor change in the soil microbial residue concentrations but a significant change in the contribution of microbial residue-C to SOC after N addition. The contribution of fungal residue-C to SOC decreased under low N addition (50 kg N ha-1 yr-1) in the tropical secondary forest (-19%), but increased under high N addition (100 kg N ha-1 yr-1) in the temperate Korean pine mixed forest (+21%). The contribution of bacterial residue-C to SOC increased under the high N addition in the subtropical Castanopsis carlesii forest (+26%) and under the low N addition in the temperate birch forest (+38%), respectively. The responses of microbial residue-C in SOC to N addition depended on the changes in soil total N concentration and fungi to bacteria ratio under N addition and climate. Taken together, these findings provide the experimental evidence that N addition diversely regulates the formation and composition of microbial-derived C in SOC in forest ecosystems.

Responses of forest ecosystems to increasing N deposition in China: A critical review.

China has been experiencing a rapid increase in nitrogen (N) deposition due to intensified anthropogenic N emissions since the late 1970s. By synthesizing experimental and observational data taken from literature, we reviewed the responses of China's forests to increasing N deposition over time, with a focus on soil biogeochemical properties and acidification, plant nutrient stoichiometry, understory biodiversity, forest growth, and carbon (C) sequestration. Nitrogen deposition generally increased soil N availability and soil N leaching and decreased soil pH in China's forests. Consequently, microbial biomass C and microbial biomass N were both decreased, especially in subtropical forests. Nitrogen deposition increased the leaf N concentration and phosphorus resorption efficiency, which might induce nutrient imbalances in the forest ecosystems. Although experimental N addition might not affect plant species richness in the overstorey, it did significantly alter species composition of understory plants. Increased N stimulated tree growth in temperate forests, but this effect was weak in subtropical and tropical forests. Soil respiration in temperate forests was non-linearly responsive to N additions, with an increase at dosages of <60 kg N ha-1 yr-1 and a decrease at dosages of >60 kg N ha-1 yr-1. However, it was consistently decreased by increased N inputs in subtropical and tropical forests. In light of future trends in the composition (e.g., reduced N vs. oxidized N) and the loads of N deposition in China, further research on the effects of N deposition on forest ecosystems will have critical implications for the management strategies of China's forests.

The response of tree growth to nitrogen and phosphorus additions in a tropical montane rainforest.

Rapid increase of global nitrogen (N) deposition has greatly altered carbon cycles and functioning of forest ecosystems. Previous studies have focused on changes in carbon dynamics of temperate and subtropical forests through N enrichment experiments; however, the effects of N deposition on tree growth remain inconsistent, especially in tropical forests. Here, we conducted a five-year N addition experiment (0 and 50kgNha-1yr-1) in a tropical montane rain forest in Hainan Island, China, to explore the effects of enhanced N deposition on growth of trees. We also set phosphorus (P) treatment (50kgPha-1yr-1) and N+P treatment (50kgNha-1yr-1+50kgPha-1yr-1) to examine potential P limitation driven by N deposition. Our results showed that N addition has not significantly influenced tree growth, while P addition significantly increased the relative growth rate of small (diameter at breast height, DBH≤10cm) and medium (10

Effects of nitrogen deposition on soil microbial communities in temperate and subtropical forests in China.

Increasing nitrogen (N) deposition has aroused large concerns because of its potential negative effects on forest ecosystems. Although microorganisms play a vital role in ecosystem carbon (C) and nutrient cycling, the effect of N deposition on soil microbiota still remains unclear. In this study, we investigated the responses of microbial biomass C (MBC) and N (MBN) and microbial community composition to 4-5years of experimentally simulated N deposition in temperate needle-leaf forests and subtropical evergreen broadleaf forests in eastern China, using chloroform fumigation extraction and phospholipid fatty acid (PLFA) methods. We found idiosyncratic effects of N addition on microbial biomass in these two types of forest ecosystems. In the subtropical forests, N addition showed a significant negative effect on microbial biomass and community composition, while the effect of N addition was not significant in the temperate forests. The N addition decreased MBC, MBN, arbuscular mycorrhizal fungi, and the F/B ratio (ratio of fungi to bacteria biomass) in the subtropical forests, likely due to a decreased soil pH and changes in the plant community composition. These results showed that microbial biomass and community composition in subtropical forests, compared with the temperate forests, were sensitive to N deposition. Our findings suggest that N deposition may have negative influence on soil microorganisms and potentially alter carbon and nutrient cycling in subtropical forests, rather than in temperate forests.

Nitrogen deposition has minor effect on soil extracellular enzyme activities in six Chinese forests.

Soil extracellular enzymes play a key role in mediating a range of forest ecosystem functions (i.e., carbon and nutrients cycling and biological productivity), particularly in the face of atmospheric N deposition that has been increasing at an unprecedented rate globally. However, most studies have focused only on surface soils in a single ecosystem. In this study, we aimed to determine whether the effect of simulated N deposition on the activities and ratios of soil enzymes changes with soil depth across six forest ecosystems in eastern China. We collected soil samples from three blocks×four soil depths (0-10cm, 10-20cm, 20-40cm and 40-60cm)×three N treatment levels (control, 50 and 100kgNha-1year-1) at each of the six forest ecosystems. We measured the activities of seven soil enzymes involved in C-, N- and P-cycling. We found that 4-5years of N addition had no significant effect on the activities and ratios of these enzymes in most cases. The interactions among N addition, site and soil depth on soil enzyme activities were not significant, except that acid phosphatase activity showed site-specific responses to N addition. Our findings suggest that the activities of soil enzymes involved in C- and N-cycling generally do not track simulated N deposition in the six forest ecosystems. Further work on plant, soil and microbial characteristics is needed to better understand the mechanisms of soil enzyme activities in response to N deposition in forest ecosystems.

Scaling of nitrogen and phosphorus across plant organs in shrubland biomes across Northern China.

Allocation of limiting resources, such as nutrients, is an important adaptation strategy for plants. Plants may allocate different nutrients within a specific organ or the same nutrient among different organs. In this study, we investigated the allocation strategies of nitrogen (N) and phosphorus (P) in leaves, stems and roots of 126 shrub species from 172 shrubland communities in Northern China using scaling analyses. Results showed that N and P have different scaling relationships among plant organs. The scaling relationships of N concentration across different plant organs tended to be allometric between leaves and non-leaf organs, and isometric between non-leaf organs. Whilst the scaling relationships of P concentration tended to be allometric between roots and non-root organs, and isometric between non-root organs. In arid environments, plant tend to have higher nutrient concentration in leaves at given root or stem nutrient concentration. Evolutionary history affected the scaling relationships of N concentration slightly, but not affected those of P concentration. Despite fairly consistent nutrients allocation strategies existed in independently evolving lineages, evolutionary history and environments still led to variations on these strategies.

A test of the generality of leaf trait relationships on the Tibetan Plateau.

Leaf mass per area (LMA), nitrogen concentration (on mass and area bases, N(mass) and N(area), respectively), photosynthetic capacity (A(mass) and A(area)) and photosynthetic nitrogen use efficiency (PNUE) are key foliar traits, but few data are available from cold, high-altitude environments. Here, we systematically measured these leaf traits in 74 species at 49 research sites on the Tibetan Plateau to examine how these traits, measured near the extremes of plant tolerance, compare with global patterns. Overall, Tibetan species had higher leaf nitrogen concentrations and photosynthetic capacities compared with a global dataset, but they had a slightly lower A(mass) at a given N(mass). These leaf trait relationships were consistent with those reported from the global dataset, with slopes of the standardized major axes A(mass)-LMA, N(mass)-LMA and A(mass)-N(mass) identical to those from the global dataset. Climate only weakly modulated leaf traits. Our data indicate that covarying sets of leaf traits are consistent across environments and biogeographic regions. Our results demonstrate functional convergence of leaf trait relationships in an extreme environment.