Extraction and identification of polyphenol from Camellia oleifera leaves using tailor-made deep eutectic solvents based on COSMO-RS design.

Camellia oleifera leaf is a rich source of polyphenols. In this study, 50 polyphenolic compounds from C. oleifera leaves was identified by UHPLC-Q-TOF-MS/MS. Accordingly, COSMO-RS was used in the design of deep eutectic solvents (DESs) to extract those polyphenols. 17 types of choline chloride (ChCl)-based DESs molecules (ChCl-acid, ChCl-sugar, ChCl-alcohol, ChCl-amine and amide) were synthetized into virtual cluster molecules with Materials Studio software. They were used to determine the activity coefficients with the standard compounds. The results showed that the amine and amide-based DESs exhibited outstanding dissolution effects. Additionally, ChCl-acetamide was selected as the solvent in response surface methodology to optimize the ultrasound-assisted DES extraction process parameters, including ultrasonic power, ultrasonic time, and liquid-solid ratio, resulting in an improved total phenolic content of 131.63 ± 0.85 mg GAE/g. This study developed a system utilizing UHPLC-Q-TOF-MS/MS to acquire specific substances required for COSMO-RS calculations.

Weakened casual feedback loops following intensive restoration efforts and climate changes in a large shallow freshwater lake.

Understanding how phytoplankton interacts with local and regional drivers as well as their feedbacks is a great challenge, and quantitative analyses of the regulating role of human activities and climate changes on these feedback loops are also limited. By using monthly monitoring dataset (2000-2017) from Lake Taihu and empirical dynamic modelling to construct causal networks, we quantified the strengths of causal feedbacks among phytoplankton, local environments, zooplankton, meteorology as well as global climate oscillation. Prevalent bidirectional causal linkages between phytoplankton biomass (chlorophyll a) and the tested drivers were found, providing holistic and quantitative evidence of the ubiquitous feedback loops. Phytoplankton biomass exhibited the highest feedbacks with total inorganic nitrogen and ammonia and the lowest with nitrate. The feedbacks between phytoplankton biomass and environmental factors from 2000 to 2017 could be classified into two groups: the local environments (e.g., nutrients, pH, transparency, zooplankton biomass)-driven enhancement loops promoting the response of the phytoplankton biomass, and the climate (e.g., wind speed)-driven regulatory loops suppressing it. The two counterbalanced groups modified the emergent macroecological patterns. Our findings revealed that the causal feedback networks loosened significantly after 2007 following nutrient loading reduction and unsuccessful biomanipulation restoration attempts by stocking carp. The strength of enhancement loops underwent marked decreases leading to reduced phytoplankton responses to the tested drivers, while the climate (decreasing wind speed, warming winter)-driven regulatory loops increased- like a tug-of-war. To counteract the self-amplifying feedback loops, the present eutrophication mitigation efforts, especially nutrient reduction, should be continued, and introduction of alternative measures to indirectly regulate the critical components (e.g., pH, Secchi depth, zooplankton biomass) of the loops would be beneficial.

Macrophage-mediated trogocytosis contributes to destroying human schistosomes in a non-susceptible rodent host, Microtus fortis.

Schistosoma parasites, causing schistosomiasis, exhibit typical host specificity in host preference. Many mammals, including humans, are susceptible to infection, while the widely distributed rodent, Microtus fortis, exhibits natural anti-schistosome characteristics. The mechanisms of host susceptibility remain poorly understood. Comparison of schistosome infection in M. fortis with the infection in laboratory mice (highly sensitive to infection) offers a good model system to investigate these mechanisms and to gain an insight into host specificity. In this study, we showed that large numbers of leukocytes attach to the surface of human schistosomes in M. fortis but not in mice. Single-cell RNA-sequencing analyses revealed that macrophages might be involved in the cell adhesion, and we further demonstrated that M. fortis macrophages could be mediated to attach and kill schistosomula with dependence on Complement component 3 (C3) and Complement receptor 3 (CR3). Importantly, we provided direct evidence that M. fortis macrophages could destroy schistosomula by trogocytosis, a previously undescribed mode for killing helminths. This process was regulated by Ca2+/NFAT signaling. These findings not only elucidate a novel anti-schistosome mechanism in M. fortis but also provide a better understanding of host parasite interactions, host specificity and the potential generation of novel strategies for schistosomiasis control.

An efficient metal-organic framework-based drug delivery platform for synergistic antibacterial activity and osteogenesis.

Bone implants for clinical application should be endowed with antibacterial activity, biocompatibility, and even osteogenesis-promoting properties. In this work, metal-organic framework (MOF) based drug delivery platform was used to modify titanium implants for improved clinical applicability. Methyl Vanillate@Zeolitic Imidazolate Framework-8 (MV@ZIF-8) was immobilized on the polydopamine (PDA) modified titanium. The sustainable release of the Zn2+ and MV causes substantial oxidative damage to Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). The increased reactive oxygen species (ROS) significantly up-regulates the expression of oxidative stress and DNA damage response genes. Meanwhile, the structural disruption of lipid membranes caused by the ROS, the damage caused by Zinc active sites and the damage accelerated by the MV are both involved in inhibiting bacterial proliferation. The up-regulated expression of the osteogenic-related genes and proteins indicated that the MV@ZIF-8 could effectively promote the osteogenic differentiation of the human bone mesenchymal stem cells (hBMSCs). RNA sequencing and Western blotting analysis revealed that the MV@ZIF-8 coating activates the canonical Wnt/ß-catenin signaling pathway through the regulation of tumor necrosis factor (TNF) pathway, thereby promoting the osteogenic differentiation of the hBMSCs. This work demonstrates a promising application of the MOF-based drug delivery platform in bone tissue engineering.

Migration deficits of the neural crest caused by CXADR triplication in a human Down syndrome stem cell model.

Down syndrome (DS) is the most common chromosomal abnormality in live-born infants and is caused by trisomy of chromosome 21. Most individuals with DS display craniofacial dysmorphology, including reduced sizes of the skull, maxilla, and mandible. However, the underlying pathogenesis remains largely unknown. Since the craniofacial skeleton is mainly formed by the neural crest, whether neural crest developmental defects are involved in the craniofacial anomalies of individuals with DS needs to be investigated. Here, we successfully derived DS-specific human induced pluripotent stem cells (hiPSCs) using a Sendai virus vector. When DS-hiPSCs were induced to differentiate into the neural crest, we found that trisomy 21 (T21) did not influence cell proliferation or apoptosis. However, the migratory ability of differentiated cells was significantly compromised, thus resulting in a substantially lower number of postmigratory cranial neural crest stem cells (NCSCs) in the DS group than in the control group. We further discovered that the migration defects could be partially attributed to the triplication of the coxsackievirus and adenovirus receptor gene (CXADR; an adhesion protein) in the DS group cells, since knockdown of CXADR substantially recovered the cell migratory ability and generation of postmigratory NCSCs in the DS group. Thus, the migratory deficits of neural crest cells may be an underlying cause of craniofacial dysmorphology in individuals with DS, which may suggest potential targets for therapeutic intervention to ameliorate craniofacial or other neural crest-related anomalies in DS.

MOF-derived CuO@ZnO modified titanium implant for synergistic antibacterial ability, osteogenesis and angiogenesis.

Surface modification of titanium implants with antibacterial, osteogenic and even angiogenic capabilities are essential to enhance their clinical applicability. Herein, metal-organic framework (MOF) derived CuO@ZnO composite was grafted onto the polydopamine (PDA) modified titanium alloy to achieve vascularized bone regeneration. The CuO@ZnO-coated titanium effectively inhibits the formation of bacterial biofilms and the sterilization rate of Staphylococcus aureus (S. aureus) reaches 99%. Benefitting from the intrinsic porous architecture of MOFs, the Zn2+ and Cu2+ could be controllably released to facilitate the production of excess intracellular reactive oxygen species (ROS) inside the bacteria, which ensures the excellent antibacterial performance of the composite coating. The CuO@ZnO-coated titanium also exhibits good cytocompatibility, effectively promotes the adhesion and proliferation of the human bone marrow mesenchymal stem cells (hBMSCs) and reduces the level of the cell apoptosis. The up-regulated expression of the osteogenesis-related genes and the superior extracellular matrix mineralization reveals that the CuO@ZnO coating possesses fantastic osteoinductive properties. In addition, the transwell and tube formation assays of the human umbilical vein endothelial cells (HUVECs) suggest the superior angiogenesis ability of the CuO@ZnO-coated titanium. The released Cu2+ stimulated the angiogenesis of the HUVECs in vitro by up-regulating the expression of the vascular endothelial growth factor (VEGF). These findings will provide new insight into the development of multifunctional titanium implants for clinical applications.

Multi-spatial scale effects of multidimensional landscape pattern on stream water nitrogen pollution in a subtropical agricultural watershed.

Multidimensional (coupled land use, soil properties, and topography) landscape effects on stream water nitrogen (N) are complex and scale-dependent. However, studies that identify critical buffer zones that explain large variations in riverine N, and estimate specific thresholds of multidimensional landscape patterns at the class level, result in a sudden changes in riverine N pollution, are still limited. Here, a new multidimensional landscape metric that combined land use, soil properties, and topography effects was applied to various riparian buffer zones and sub-watershed scales, and their relationships to riverine N levels were investigated. We used stream water ammonium-N, nitrate-N, and total-N concentrations datasets, from 2010 to 2017, in the nine subtropical sub-watersheds in China. The results of model selection and model averaging in ordinary least squares regressions, indicated that the riparian buffer zone with widths of 400 m, had more pronounced influence on water NH4-N and TN levels than at other scales. Within the 400 m buffer zone, the key landscape metrics for NH4-N, NO3-N and TN concentrations in stream water were different, and explained up to 43.35%-76.55% (adjusted R2) of the total variation in river N levels. When ENN_MNClass17 below 39-56 m, PDClass8 above 4.63-6.55 n/km2, PLANDClass27 above 23-29%, and CONTIG_MNClass42 below 0.35-0.37% within the 400 m buffer zone, riverine NH4-N and TN would be abruptly increased. This study provided practical ideas for regulation regarding landscape management linked to watershed structure, and identified reference thresholds for multidimensional landscape metrics, which should help reduce riverine N pollution in subtropical China.

Influence path identification of topography, soil, hydrology and landscape on phosphorus buffering capacity in typical agricultural catchments in central subtropical China.

The catchment phosphorus buffering capacity (PBF) determines the pressure-state-response relationship between anthropogenic P inputs and aquatic ecosystems at a catchment scale, and is affected by biogeochemical, hydrological, and ecological catchment characteristics. However, the complex relationship between these catchment characteristic factors and their impact pathways on PBF remains ambiguous, leading to large uncertainty in balancing agricultural productivity and water conservation via improving BF through management practices. In this study, the short-term buffering index, calculated from net anthropogenic P input and riverine P exports, was used to quantify the spatiotemporal variations in PBF in source agricultural catchments in the Dongting Lake basin. Partial least squares structural equation modeling was used to investigate the relationship between the PBF and the catchment characteristics. The results indicate that catchment PBF was directly determined by soil properties and hydrological conditions, while landscape patterns significantly mediated the effects of topography on soil and hydrology. Considering the pathway preferences of the model, landscape patterns could be the priority for characterizing and regulating PBF. According to a change-point analysis, the probability of PBF weakening increases dramatically when the proportion of farmland (Farm%) > 24.6%, degree of patch interspersion (Contagion index) < 64.5%, and Perimeter-Area Ratio Distribution (PARA) > 348.7. These findings provide new insights into catchment buffering mechanisms and can be used to promote the simultaneous achievement of agricultural production and environmental conservation goals.

Landscape patterns of catchment and land-use regulate legacy phosphorus releases in subtropical mixed agricultural and woodland catchments.

Landscape composition and configuration determine the exchange of matter and energy among different landscape patches and may affect riverine phosphorus (P) exports derived from watershed legacy sources. However, a lack of understanding of landscape pattern effects on legacy P releases has yielded large uncertainties in mitigating watershed water quality using management practices or landscape planning. This study revealed the significance of legacy effect in the headwater catchments through the time-lag response of the long-term trend of river P exports to the change of net anthropogenic P input (NAPI). By constructing empirical statistical models that incorporated NAPI, hydroclimatic, terrain factors, soil chemical properties, and land use variables, the sources of annual riverine total phosphorus (TP) and dissolved inorganic phosphorus (DIP) exports were divided into current annual NAPI input and legacy sources inputs. The model estimations indicated that the contribution of legacy sources to riverine TP exports was 0.33-1.12 kg ha-1 yr-1 (50.7-82.8%), which was significantly higher than the contribution to DIP exports (0.18-0.49 kg ha-1 yr-1, 42.4-81.4%) in 2012-2017. Redundancy analysis (RDA) and variance partitioning analysis (VPA) methods were used to quantify the relative contribution of landscape patterns, soil P content, and terrain factors to legacy P releases. Results revealed that the relative contribution of the landscape composition and configuration to the total variations of legacy P releases was greater than that of the soil P and terrain factors. For different land use patches, a large area of woodland with a high aggregation degree and a large area of ponds with multiple net structures may significantly alleviate legacy P releases. In contrast, the legacy P releases were significantly positively associated with highly aggregated agricultural, tea plantation, and residential patches. This study provides theoretical support for strategies aiming to control legacy P from the perspective of landscape planning.

Linking human activities and global climatic oscillation to phytoplankton dynamics in a subtropical lake.

Human activities and climate change are two major stressors affecting lake ecosystems as well as phytoplankton communities worldwide. However, how the temporal dynamics of phytoplankton are directly or indirectly linked to anthropogenic activities and climatic oscillation remains unclear. We assessed the annual trends (1988-2018) in phytoplankton abundance (PA) in Lake Dongting, China and related it to five groups of variables characterizing human activities, global climate oscillation, water nutrients, hydrology, and meteorology. We found a significant increase in PA, urbanization (Upop), total nitrogen (TN), fertilizer application (FA), number of summer days (SU), and the warm speed duration index (WSDI) and a significant decrease in the water discharge of three inlets (TIWD) and the sediment discharge of three inlets (TISD) and four tributaries (FTSD) and the net sediment deposition (NSD). However, no significant annual trends were observed for the number of rainstorm days (R50mm), the simple precipitation intensity index (SDII) and yearly anomalies of El Niño-Southern oscillation events (ENSOi). Cross-correlation Function analyses demonstrated that the operation of the Three George Dam (TGD) strengthened the effects of hydrology, rainfall patterns and ENSOi on phytoplankton, but strongly weakened the association between water nutrients, human activities and phytoplankton abundance. Path analysis revealed that TP, TN, FA, R50 mm as well as WSDI had a direct positive effect on PA, while a direct negative effect was found for ENSOi, NSD and TISD. Human activities (Upop and FA), warming (WSDI and SU), and rainfall patterns (SDII and R50 mm) exerted indirect controls on phytoplankton through changes in water nutrients and hydrology. Climate change (ENSOi) had a direct effect on PA, but also showed twelve indirect pathways via changes in hydrology and meteorology (both positive and negative effects were found). Overall, meteorology contributed most markedly to the variations of PA (29.3%), followed by hydrology (25.3%), human activities (24%), water nutrients (10.5%), and ENSOi (1.9%). Our results highlight a strongly causal connection between human activities as well as global climate change and phytoplankton and the benefits of considering multiple environmental drivers in determining the temporal dynamics of lake biotic communities.

From landscape perspective to determine joint effect of land use, soil, and topography on seasonal stream water quality in subtropical agricultural catchments.

Understanding the relationship between integrated landscape patterns (coupled land use, soil properties, and topography) and stream water quality in different seasons promotes appropriate landscape planning. However, this relationship is unclear. Here, water quality nitrogen (N) and phosphorus (P) levels and the integrated landscape patterns were investigated in ten Chinese subtropical catchments during 2010-2017, using the least absolute shrinkage and selection operator (LASSO) regressions method and redundancy analysis (RDA). The results suggested that stream water N and P levels were significantly higher in the fallow season than in the rice-growing season (p < 0.05). The N and P levels in the rice-growing season were elevated with the increasing area proportions of tea fields in Ultisols on the medium slope (16.06-28.02°), and larger isolation, diversity, and geometric complexity of landscape patches, but decreased with the increasing area proportions of forests in Ultisols on the steep slope (28.02-80.30°) and interspersion of landscape patches. Stream water N and P levels were more likely influenced by landscape configuration in the rainy rice-growing season, mainly due to the rapid velocities and high quantities of surface flow strengthening landscape configuration effects on the N and P migration and exchange. In the fallow season, the N and P levels were heightened with the increasing area proportions of tea fields, residential areas, and paddy fields in Ultisols on the relative flat (0-16.06°) and medium slopes, and larger isolation of landscape, and could be greatly reduced if the area proportions of forests in Ultisols on the steep slope is increased. The N and P levels were more likely determined by landscape composition in the dry fallow season, associated with the slow and poor subsurface and underground hydrological flows. Therefore, the results promoted reasonable landscape management in different seasons and suitable soil and topography conditions for improving stream water quality.

Characterization and Therapeutic Application of Mesenchymal Stem Cells with Neuromesodermal Origin from Human Pluripotent Stem Cells.

Rationale: Mesenchymal stem cells (MSC) hold great promise in the treatment of various diseases including autoimmune diseases, inflammatory diseases, etc., due to their pleiotropic properties. However, largely incongruent data were obtained from different MSC-based clinical trials, which may be partially due to functional heterogeneity among MSC. Here, we attempt to derive homogeneous mesenchymal stem cells with neuromesodermal origin from human pluripotent stem cells (hPSC) and evaluate their functional properties. Methods: Growth factors and/or small molecules were used for the differentiation of human pluripotent stem cells (hPSC) into neuromesodermal progenitors (NMP), which were then cultured in animal component-free and serum-free induction medium for the derivation and long-term expansion of MSC. The resulted NMP-MSC were detailed characterized by analyzing their surface marker expression, proliferation, migration, multipotency, immunomodulatory activity and global gene expression profile. Moreover, the in vivo therapeutic potential of NMP-MSC was detected in a mouse model of contact hypersensitivity (CHS). Results: We demonstrate that NMP-MSC express posterior HOX genes and exhibit characteristics similar to those of bone marrow MSC (BMSC), and NMP-MSC derived from different hPSC lines show high level of similarity in global gene expression profiles. More importantly, NMP-MSC display much stronger immunomodulatory activity than BMSC in vitro and in vivo, as revealed by decreased inflammatory cell infiltration and diminished production of pro-inflammatory cytokines in inflamed tissue of CHS models. Conclusion: Our results identify NMP as a new source of MSC and suggest that functional and homogeneous NMP-MSC could serve as a candidate for MSC-based therapies.

Response of regional agricultural soil phosphorus status to net anthropogenic phosphorus input (NAPI) determined by soil pH value and organic matter content in subtropical China.

Exploring the relationship between net anthropogenic phosphorus input (NAPI) and soil available P (SAP) content could inform applied issues related to environmental quality and agronomic productivity and increase our knowledge of element biogeochemical cycles. Here, the NAPI was estimated and the SAP content determined in eight counties in subtropical China from 1980 to 2010. It is suggested that the NAPI ranging 318-924 km-2 yr-1 in 1980 had increased substantially to 865-3601 km-2 yr-1 in 2010 across the eight counties, in which the P fertilizer application was estimated to represent the largest individual source of NAPI, accounting for an average of 36.1-74.6% of the NAPI. The NAPI in agricultural land (NAPIa) was the largest component of the NAPI, and 60.7-77.1% of the NAPIa accumulated in the upper 20 cm layer of agricultural soils, which significantly increased soil total-P (TP) and SAP contents. The increases in SAP, resulting from 10,000 kg P km-2 of the NAPIa (IOPNAPI), were estimated to be 1.61-4.36 mg P kg-1 in the counties. Both the correlation and variation partitioning analyses (VPAs) suggested that the soil pH and organic matter content (SOM) were the most important factors influencing the variations of IOPNAPI (determination coefficient: 72.5%). Therefore, the contribution of soil pH and SOM should be considered in enriching soil SAP levels and implementing optimal P management strategies to improving the agronomic effectiveness of P fertilization and further reduce the environmental risk of P loss in subtropical region.

[Regional differences of inputs of organic matter and chemical fertilizer in South Central China].

This article analyzed the inputs of organic matter and chemical fertilizer in the cropland of South Central China, i.e., Hunan, Hubei, Guangdong and Guangxi, and then calculated the budgets of nitrogen (N), phosphorus (P) and potassium (K), based on the data from field investigations and peasant household surveys in the four provinces. The results showed that total amounts of organic matter inputs in the four provinces was ranked as follow: 8993 kg · hm(-2) in Guangxi, 6390 kg · hm(-2) in Hunan, 5012 kg · hm(-2) in Hubei, 4630 kg · hm(-2) in Guangdong, and average NPK inputs in the four provinces were ranked as follow: 777.5 kg · hm(-2) in Guangxi, 501.6 kg · hm(-2) in Hunan, 486.4 kg · hm(-2) in Hubei, 340.4 kg · hm(-2) in Guangdong. The N and P input surpluses were greatest in Guangxi (67.2% and 99.0% as for N and P, respectively) , followed by Hunan (33.2% and 50.8%), Hubei (11.8% and 11.0%), and Guangdong (7.8% and 30.0%). However, K input was deficient in Hunan, Hubei, and Guangdong (6.6%, 18.7% and 12.4%), but surplus in Guangxi (19.5%).