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Peatlands play a crucial role in mediating the emissions of methane through active biogeochemical cycling of accumulated carbon driven by methane-metabolizing microorganisms; meanwhile, they serve as vital archives of atmospheric heavy metal deposition. Despite many edaphic factors confirmed as determinants to modulate the structure of methanotrophic and methanogenic communities, recognition of interactions among them is limited. In this study, peat soils were collected from Dajiuhu peatland to assess the presence of heavy metals, and methanotrophs and methanogens were investigated via high-throughput sequencing for functional genes mcrA and pmoA. Further analyses of the correlations between methane-related functional groups were conducted. The results demonstrated that both methane-metabolizing microorganisms and heavy metals have prominent vertical heterogeneity upward and downward along the depth of 20 cm. Pb, Cd, and Hg strongly correlated with methanotrophs and methanogens across all seasons and depths, serving as forceful factors in structural variations of methanogenic and methanotrophic communities. Particularly, Pb, Cd, and Hg were identified as excessive elements in Dajiuhu peatland. Furthermore, seasonal variations of networks among methane-related functional groups and environmental factors significantly affected the changes of methane fluxes across different seasons. Concretely, the complicated interactions were detrimental to methane emissions in the Dajiuhu peatland, leading to the minimum methane emissions in winter. Our study identified the key heavy metals affecting the composition of methane-metabolizing microorganisms and linkages between seasonal variations of methane emissions and interaction among heavy metals and methane-metabolizing microorganisms, which provided much new reference and theoretical basis for integrated management of natural peatlands.

Wetland ecosystems are the most important natural sources of methane (CH4), with periodical changes of CH4 fluxes observed. Methanogens and methanotrophs are considered as the key factors affecting CH4 fluxes in wetlands. However, the relationship between methanogens and methanotrophs was still unclear. To close this gap, samples at four depths of soil in the Dajiuhu subalpine peatland in January, April, July, and October 2019 were collected and CH4 fluxes data from the eddy covariance (EC) system was acquired. The phylogenetic molecular ecological networks (pMENs) analysis was used to identify keystone species and the seasonal variations of co-occurrence patterns of methanogenic and methanotrophic communities. The results indicated that the seasonal variations of interactions between methanogenic and methanotrophic communities contributed to CH4 emissions in wetland. The keystone species discerned by network analysis also showed their importance in mediating CH4 fluxes. CH4 emissions in wetland were lowest in spring, during this period, most complex interactions between microbes were observed, with the intense competition among methanogens while methanotrophs represented better cooperation. Reverse patterns manifested in summer, when the highest CH4 flux was observed. Meanwhile, Methanoregula formicica was proven negatively correlated with CH4 fluxes that occupied largest ecological niches in the spring network, and Methanocella arvoryzae, as well as Methylocystaceae had positive correlations with CH4 fluxes survived better in the microbial community in summer. In addition, soil temperature and nitrogen were regarded as significant environmental factors to CH4 fluxes. This study explained the seasonal patterns and microbial driving mechanisms of CH4 emissions in wetlands.

The rapid development of modern society has resulted in discharge of large, heavy metal quantities into wetlands that have been continuously accumulating, causing severe pollution. Dajiuhu, located in the Shennongjia Forest District of Hubei Province in China, is a wetland of significant value internationally, serving as a model wetland ecosystem with heightened scientific research value. In this study, 27 surface sediment samples from nine sub-lakes in Dajiuhu were collected in August 2020. The concentrations of Cd, Cr, Cu, Ni, Pb, and Zn in the sediments were determined. The heavy metal occurrence and speciation characteristics were analyzed by an improved BCR (European Community Bureau of Reference) extraction method. Four methods were used to evaluate heavy metals' pollution degree and ecological risk. The possible source of heavy metals was inferred using correlation analysis and principal component analysis. The heavy metal content in the lake sediments of Dajiuhu wetland was from the highest to the lowest concentration as follows: Zn [Formula: see text] Cr [Formula: see text] Ni [Formula: see text] Pb [Formula: see text] Cu [Formula: see text] Cd. The average Cd content exceeded the national nature reserve threshold values, while the other heavy metals measured were below their respective threshold values. However, due to the occurrence of Pb and Cd in different forms, they still pose certain pollution and ecological risk to the lake wetlands. On the other hand, Zn, Cr, Ni, and Cu do not pose an ecological risk in the lakes of the Dajiuhu wetland. The spatial distribution of heavy metal content in the nine sub-lakes did vary significantly. Regarding the heavy metal sources in the lake sediments, Ni, Cr, and Cu originate from natural factors, and Cd and Pb have mainly anthropogenic origins. In contrast, Zn has both natural and anthropogenic origins. This study provides further insights into the study of heavy metal pollution in lake wetlands. It provides a framework and a direction for managing heavy metal pollution in the Dajiuhu wetland.

Renal cell carcinoma (RCC) is one of the most drug-resistant malignancies, and an effective therapy is lacking for metastatic RCC. Anisomycin is known to inhibit protein synthesis and induce ribotoxic stress. The aim of this study was to explore whether anisomycin enhances the cytotoxic effects of mapatumumab, a human agonistic monoclonal antibody specific for death receptor 4 (DR4), in human RCC cells. We examined the cytotoxicity of anisomycin alone and in combination with mapatumumab in human RCC cell lines and primary RCC cell cultures. RCC cells treated with anisomycin showed cytotoxicity in a dose-dependent manner. Anisomyin in combination with mapatumumab showed a synergistic effect not only in two human RCC cell lines but also in five primary RCC cell cultures. The synergy between anisomycin and mapatumumab for cytotoxicity was also observed for apoptosis. Interestingly, anisomycin significantly increased DR4 expression at both the mRNA and the protein level. Furthermore, the combination-induced cytotoxicity was significantly suppressed by a human recombinant DR4:Fc chimeric protein. The combination of anisomycin and mapatumumab also enhanced the activity of caspases 8 and 3, the downstream molecules of death receptors. These findings indicate that anisomycin sensitizes RCC cells to DR4-mediated apoptosis through the induction of DR4, suggesting that combinational treatment with anisomycin and mapatumumab might represent a novel therapeutic strategy for the treatment of RCC.