Ubiquitination and regulation of Smad7 in the TGF-ß1/Smad signaling of aristolochic acid nephropathy.

Aristolochic acid I (AAI) affects TGF-ß1/Smad signaling, which causes AA nephropathy (AAN), but the mechanisms are not fully understood. We aimed to clarify whether Arkadia and UCH37 participate in TGF-ß1/Smad signaling via Smad7, and the regulatory mechanisms of Smad7. One side, mice and cultured mouse renal tubular epithelial cells (RTECs) were treated with various AAI doses and concentrations, respectively; on the other side, RTECs were transfected with small interfering RNA (siRNA) expression vectors against Arkadia and UCH37 and then treated with 10 µg/ml AAI. And then detect the mRNA and protein levels of Smad7, UCH37, Arkadia and any other relative factors by RT-PCR and Western blotting. In kidney tissues and RTECs, the mRNA and protein levels of Smad7 decreased with increasing AAI doses concentrations by real-time PCR and Western blotting, whereas those of Arkadia, UCH37, Smad2, Smad3 and TßRI increased. Cells transfected with the Arkadia siRNA expression vector showed reduced mRNA and protein levels of vimentin, α-SMA, Smad2, Smad3 and TßRI after AAI treatment, while those of CK18 and Smad7 increased compared with those of untransfected RTECs. Conversely, cells transfected with the UCH37 siRNA expression vector showed the opposite effect on analyzed signaling molecules after AAI treatment. Arkadia and UCH37 participate in TGF-ß1/Smad signaling-mediated renal fibrosis, and Smad7 blocks TGF-ß1 signaling by inhibiting Smad2/Smad3 phosphorylation and enhancing the degradation of TßRI.

Expression of histone deacetylase-1 and p300 in aristolochic acid nephropathy models.

Aristolochic acid nephropathy (AAN) is mainly caused by aristolochic acid I (AAI), but the actual mechanism is still uncertain. The current study explored the correlation among the expression of Smad7, p300, histone deacetylase-1 (HDAC1) and the development of AAN using transmission electron microscopy (TEM), RT-PCR, and western blotting in the AAN mouse model and in the AAN cell model. TEM revealed that the renal tubular epithelial cells from the AAI-treated mice presented organelle damages and nuclear deformation. We found that a certain dose of AAI caused renal fibrosis and induced renal tubular epithelial cells to differentiate into myofibroblasts. There was a gradual increase in the expression of HDAC1 mRNA and protein observed using RT-PCR and western blotting in the AAN cell model compared with the control group. Gradual decrease in the expression of Smad7 and p300 mRNA and protein was revealed in the AAN mouse and cell models compared with the control group. These results suggest that AAI dose dependently contributed to the development of AAN, and HDAC1 and p300 participate in the modulation of TGF-ß/Smad pathway-mediated renal interstitial fibrosis.

The impact of technological innovation on the green digital economy and development strategies.

To investigate the interplay among technological innovation, industrial structure, production methodologies, economic growth, and environmental consequences within the paradigm of a green economy and to put forth strategies for sustainable development, this study scrutinizes the limitations inherent in conventional deep learning networks. Firstly, this study analyzes the limitations and optimization strategies of multi-layer perceptron (MLP) networks under the background of the green economy. Secondly, the MLP network model is optimized, and the dynamic analysis of the impact of technological innovation on the digital economy is discussed. Finally, the effectiveness of the optimization model is verified by experiments. Moreover, a sustainable development strategy based on dynamic analysis is also proposed. The experimental results reveal that, in comparison to traditional Linear Regression (LR), Decision Tree (DT), Random Forest (RF), Support Vector Machine (SVM), and Naive Bayes (NB) models, the optimized model in this study demonstrates improved performance across various metrics. With a sample size of 500, the optimized model achieves a prediction accuracy of 97.2% for forecasting future trends, representing an average increase of 14.6%. Precision reaches 95.4%, reflecting an average enhancement of 19.2%, while sensitivity attains 84.1%, with an average improvement of 11.8%. The mean absolute error is only 1.16, exhibiting a 1.4 reduction compared to traditional models and confirming the effectiveness of the optimized model in prediction. In the examination of changes in industrial structure using 2020 data to forecast the output value of traditional and green industries in 2030, it is observed that the output value of traditional industries is anticipated to decrease, with an average decline of 11.4 billion yuan. Conversely, propelled by the development of the digital economy, the output value of green industries is expected to increase, with an average growth of 23.4 billion yuan. This shift in industrial structure aligns with the principles and trends of the green economy, further promoting sustainable development. In the study of innovative production methods, the green industry has achieved an increase in output and significantly enhanced production efficiency, showing an average growth of 2.135 million tons compared to the average in 2020. Consequently, this study highlights the dynamic impact of technological innovation on the digital economy and its crucial role within the context of a green economy. It holds certain reference significance for research on the dynamic effects of the digital economy under technological innovation.

Deciphering Microbial Community and Nitrogen Fixation in the Legume Rhizosphere.

Nitrogen is the most limiting factor in crop production. Legumes establish a symbiotic relationship with rhizobia and enhance nitrogen fixation. We analyzed 1,624 rhizosphere 16S rRNA gene samples and 113 rhizosphere metagenomic samples from three typical legumes and three non-legumes. The rhizosphere microbial community of the legumes had low diversity and was enriched with nitrogen-cycling bacteria (Sphingomonadaceae, Xanthobacteraceae, Rhizobiaceae, and Bacillaceae). Furthermore, the rhizosphere microbiota of legumes exhibited a high abundance of nitrogen-fixing genes, reflecting a stronger nitrogen-fixing potential, and Streptomycetaceae and Nocardioidaceae were the predominant nitrogen-fixing bacteria. We also identified helper bacteria and confirmed through metadata analysis and a pot experiment that the synthesis of riboflavin by helper bacteria is the key factor in promoting nitrogen fixation. Our study emphasizes that the construction of synthetic communities of nitrogen-fixing bacteria and helper bacteria is crucial for the development of efficient nitrogen-fixing microbial fertilizers.

Combined effects of azoxystrobin and oxytetracycline on rhizosphere microbiota of Arabidopsis thaliana.

The rhizosphere is one of the key determinants of plant health and productivity. Mixtures of pesticides are commonly used in intensified agriculture. However, the combined mechanisms underlying their impacts on soil microbiota remain unknown. The present study revealed that the rhizosphere microbiota was more sensitive to azoxystrobin and oxytetracycline, two commonly used pesticides, than was the microbiota present in bulk soil. Moreover, the rhizosphere microbiota enhanced network complexity and stability and increased carbohydrate metabolism and xenobiotic biodegradation as well as the expression of metabolic genes involved in defence against pesticide stress. Co-exposure to azoxystrobin and oxytetracycline had antagonistic effects on Arabidopsis thaliana growth and soil microbial variation by recruiting organic-degrading bacteria and regulating ABC transporters to reduce pesticide uptake. Our study explored the composition and function of soil microorganisms through amplicon sequencing and metagenomic approaches, providing comprehensive insights into the synergistic effect of plants and rhizosphere microbiota on pesticides and contributing to our understanding of the ecological risks associated with pesticide use.

Seismic Performance of Drained Piles in Layered Soils.

The provision of drains to geotechnical elements subjected to strong ground motion can reduce the magnitude of shaking-induced excess pore pressure and the corresponding loss of soil stiffness and strength. A series of shaking table tests were conducted within layered soil models to investigate the effectiveness of drained piles to reduce the liquefaction hazard in and near pile-improved ground. The effect of the number of drains per pile and the orientation of the drains relative to the direction of shaking were evaluated in consideration of the volume of porewater discharged, the magnitude of excess pore pressure generated, and the amount of de-amplification in the ground's motion. The following main conclusions can be drawn from this study. Single, isolated piles and a group of drained piles were tested in three series of shake table tests. Relative to conventional piles, the drained piles exhibited improved performance with regard to the generation and dissipation of excess pore pressure and stiffness of the surrounding soil, with increases in performance correlated with increases in the discharge capacity of the drained pile. The acceleration time histories observed within the pile-improved soil indicated a coupling of the rate and magnitude of porewater discharge, excess pore pressure generated, and de-amplification of strong ground motion. The amount of de-amplification reduced with increases in the number of drains per pile and corresponding reductions in excess pore pressure. The improved performance should prove helpful in the presence of sloping ground characterized with low-permeability soil layers that inhibit the dissipation of pore pressure and have demonstrated the significant potential for post-shaking slope deformation.

Shaping effects of rice, wheat, maize, and soybean seedlings on their rhizosphere microbial community.

The rhizosphere microbiome plays critical roles in plant growth and is an important interface for resource exchange between plants and the soil environment. Crops at various growing stages, especially the seedling stage, have strong shaping effects on the rhizosphere microbial community, and such community reconstruction will positively feed back to the plant growth. In the present study, we analyzed the variations of bacterial and fungal communities in the rhizosphere of four crop species: rice, soybean, maize, and wheat during successive cultivations (three repeats for the seedling stages) using 16S rRNA gene and internal transcribed spacer (ITS) high-throughput sequencing. We found that the relative abundances of specific microorganisms decreased after different cultivation times, e.g., Sphingomonas, Pseudomonas, Rhodanobacter, and Caulobacter, which have been reported as plant-growth beneficial bacteria. The relative abundances of potential plant pathogenic fungi Myrothecium and Ascochyta increased with the successive cultivation times. The co-occurrence network analysis showed that the bacterial and fungal communities under maize were much more stable than those under rice, soybean, and wheat. The present study explored the characteristics of bacteria and fungi in crop seedling rhizosphere and indicated that the characteristics of indigenous soil flora might determine the plant growth status. Further study will focus on the use of the critical microorganisms to control the growth and yield of specific crops.

Understanding the ecological effects of the fungicide difenoconazole on soil and Enchytraeus crypticus gut microbiome.

Increasing knowledge of the impacts of pesticides on soil ecological communities is fundamental to a comprehensive understanding of the functional changes in the global agroecosystem industry. In this study, we examined microbial community shifts in the gut of the soil-dwelling organism Enchytraeus crypticus and functional shifts in the soil microbiome (bacteria and viruses) after 21 d of exposure to difenoconazole, one of the main fungicides in intensified agriculture. Our results demonstrated reduced body weight and increased oxidative stress levels of E. crypticus under difenoconazole treatment. Meanwhile, difenoconazole not only altered the composition and structure of the gut microbial community, but also interfered with the soil-soil fauna microecology stability by impairing the abundance of beneficial bacteria. Using soil metagenomics, we revealed that bacterial genes encoding detoxification and viruses encoding carbon cycle genes exhibited a dependent enrichment in the toxicity of pesticides via metabolism. Taken together, these findings advance the understanding of the ecotoxicological impact of residual difenoconazole on the soil-soil fauna micro-ecology, and the ecological importance of virus-encoded auxiliary metabolic genes under pesticide stress.

Interlaminar Shear Characteristics of Typical Polyurethane Mixture Pavement.

Polyurethane (PU) can be used as a road material binder, and its mechanical properties, durability, temperature stability, and other road performance metrics are good. However, the interlayer bonding between PU mixtures and asphalt mixtures is poor. The influence of the pavement structure, interlayer treatment scheme, load, and environmental factors on the interlayer shear characteristics of PU mixture composite pavement is analysed. Further, dynamic modulus, Hamburg rutting, accelerated loading, and inclined shear tests were conducted, and the typical PU mixture pavement shear stress was calculated. The interlaminar shear stress of double layer PU mixture pavement, polyurethane-asphalt composite pavement, and typical asphalt pavement were calculated. The results showed that the PU mixture has a low rutting deformation rate, stable mechanical properties, and strong resistance to the coupled action of temperature, water, and loading. The double-layer PU mixture structure has good water-temperature stability and fatigue resistance; however, freeze-thaw and accelerated loading cause great damage to the double-layer PU mixture structure. The residual shear strength ratio after freeze-thaw cycles and accelerated loading is only 50.3% and 35.6%, respectively, while the influence on the double-layer asphalt mixture structure is less. The theoretical calculation results of different pavement structures show that when the temperature increases from 10 °C to 50 °C, the interlaminar shear stress of polyurethane-asphalt composite pavement increases by about 20%. Additionally, the shear stress of pavement PU mixture pavement and typical asphalt pavement is mainly affected by load, and the temperature changes have an obvious effect on the interlayer shear stress of polyurethane-asphalt composite pavement. The calculated maximum shear stress of the three pavement structures with different working conditions is less than the interlaminar shear strength measured by the inclined shear test, indicating that the interlaminar treatment scheme of composite specimens can meet the shear resistance requirements of the three typical pavement structure types.

Development of a Potentially New Algaecide for Controlling Harmful Cyanobacteria Blooms Which is Ecologically Safe and Selective.

Harmful cyanobacterial blooms (HCBs) caused by Microcystis aeruginosa are of great concern as they negatively affect the aquatic environment and human health. Chemical methods could rapidly eradicate HCBs and have been used for many decades. However, many chemical reagents are not recommended to eliminate HCBs in the long term, given the possible destructive and toxic effects of the chemicals employed on non-target aquatic organisms. We developed a new algaecide, 2-((1,3,4-thiadiazol-2-yl)thio)-N-(4-chlorophenyl) acetamide (Q2), to control harmful cyanobacteria while being environmentally friendly and selective. In our study, Q2 effectively inhibited cyanobacterial growth, especially of M. aeruginosa, but did not affect eukaryotic algae in test concentrations. A critical mechanism was revealed by transcriptome and metagenomic results showing that Q2 affects multiple cellular targets of cyanobacteria for HCB control, including the destruction of organelles, damage in the photosynthesis center, as well as inhibition of gas vesicle growth, and these changes can be highly relevant to the decrease of quorum-sensing functional KEGG pathways. Furthermore, Q2 did not affect the microbial composition and could recover the disrupted aquatic functional pathways in a short period. This is different from the impact on ecosystem functioning of the traditionally used harmful algaecide diuron. All these results verified that Q2 could be friendly to the aquatic environment, providing a new directional choice in managing HCBs in the future.

New insights into the oleate flotation response of feldspar particles of different sizes: Anisotropic adsorption model.

The anisotropic adsorption of sodium oleate (NaOL) on feldspar surfaces was investigated to elucidate the different flotation properties of feldspar particles of four different size ranges. Microflotation experiments showed that the feldspar flotation recovery of particles with sizes spanning different ranges decreased in the order 0-19>19-38>45-75>38-45µm. Zeta potential and FTIR measurements showed that NaOL was chemically adsorbed on the Al sites of the feldspar surface. The anisotropic surface energies and broken bond densities estimated by density functional theory calculations showed that, although feldspar mostly exposed (010) and (001) surfaces, only the (001) surfaces contained the Al sites needed for NaOL adsorption. The interaction energies calculated by molecular dynamics simulations confirmed the more favorable NaOL adsorption on (001) than (010) surfaces, which may represent the main cause for the anisotropic NaOL adsorption on feldspar particles of different sizes. SEM measurements showed that the main exposed surfaces on coarse and fine feldspar particles were the side (010) and basal (001) ones, respectively. A higher fraction of Al-rich (001) surfaces is exposed on fine feldspar particles, resulting in better floatability compared with coarse particles. XPS and adsorption measurements confirmed that the Al content on the feldspar surface varied with the particle size, explaining the different NaOL flotation of feldspar particles of different sizes. Therefore, the present results suggest that coarsely ground ore should be used for the separation of feldspar gangue minerals. Further improvements in the flotation separation of feldspar from associated valuable minerals can be achieved through selective comminution or grinding processes favoring the exposure of (010) surfaces.

[Cashmere goat bacterial artificial chromosome recombination and cell transfection system].

The Cashmere goat is mainly used to produce cashmere, which is very popular for its delicate fiber, luscious softness and natural excellent warm property. Keratin associated protein (KAP) and bone morphogenetic protein (BMP) of the Cashmere goat play an important role in the proliferation and development of cashmere fiber follicle cells. Bacterial artificial chromosome containing kap6.3, kap8.1 and bmp4 genes were used to increase the production and quality of Cashmere. First, we constructed bacterial artificial chromosomes by homology recombination. Then Tol2 transposon was inserted into bacterial artificial chromosomes that were then transfected into Cashmere goat fibroblasts by Amaxa Nucleofector technology according to the manufacture's instructions. We successfully constructed the BAC-Tol2 vectors containing target genes. Each vector contained egfp report gene with UBC promoter, Neomycin resistant gene for cell screening and two loxp elements for resistance removing after transfected into cells. The bacterial artificial chromosome-Tol2 vectors showed a high efficiency of transfection that can reach 1% to 6% with a highest efficiency of 10%. We also obtained Cashmere goat fibroblasts integrated exogenous genes (kap6.3, kap8.1 and bmp4) preparing for the clone of Cashmere goat in the future. Our research demonstrates that the insertion of Tol2 transposons into bacterial artificial chromosomes improves the transfection efficiency and accuracy of bacterial artificial chromosome error-free recombination.

Transgenesis of Tol2-mediated seamlessly constructed BAC mammary gland expression vectors in Mus musculus.

Bacterial artificial chromosomes (BACs) are vectors that are capable of carrying gene fragments of up to 300 kb in size, and in theory, harbor cis-regulatory elements that are necessary for the expression of specific genes. Therefore, BACs can effectively alleviate or even eliminate the position effect induced by gene-integration, rendering these as ideal expression vectors of exogenous genes. However, the number of relevant studies involving BACs as vectors of exogenous genes are limited. In the present study, we converted the BAC regulatory region of the Mus musculus Wap gene into a mammary gland-specific expression vector. Using the galK-based positive-negative selection method, we seamlessly replaced the Wap gene in a BAC with Homo sapiens GPX3, MT2, and Luc genes while keeping the original mammary gland-specific regulatory sequence intact, without introducing any extra sequences (Loxp/Frt). To improve the efficiency of creating BAC transgenic mice, we used a Tol2 transposon system optimized for mammalian codons and eliminated 100 kb of sequence from the BAC 5' end (173 kb), which resulted in an 8.5% rate of successful gene transmission via pronuclear injection. The results of the present study indicate that seamlessly constructed BAC expression vectors can be used for the transmission of the GPX3 gene.

Clinical significance of transient receptor potential vanilloid 2 expression in human hepatocellular carcinoma.

Transient receptor potential vanilloid 2 (TRPV2), one of the members of TRP (transient receptor potential) superfamily of ion channels, has been suggested to contribute to pain associated with inflammation or neuropathy. To investigate its role in hepatocarcinogenesis, we examined the expression of TRPV2 in human hepatocellular carcinoma (HCC) samples and analyzed the association of TRPV2 expression with its clinical significance. TRPV2 expression in 55 HCC patients was examined by immunohistochemistry, and the correlation between TRPV2 levels and clinicopathologic parameters was analyzed. Thirteen paired HCC specimens and their nontumor counterparts were investigated by quantitative real-time polymerase chain reaction (RT-PCR) and Western blotting, respectively. Quantitative RT-PCR and Western blotting analysis revealed that expression of TRPV2 at both the mRNA and protein levels were increased in cirrhotic livers compared with chronic hepatitis, whereas that also occurred in moderately and well-differentiated tumors compared with that of poorly differentiated tumors. Immunohistochemistry of the 55 HCC samples showed that the expression of TRPV2 increased when going from normal liver or chronic hepatitis to cirrhosis. Increased TRPV2 expression was observed in tissues of liver cirrhosis (31/37, 83.8%). In HCC, increased expression of TRPV2 was identified in 16/55 (29%) cases. Clinicopathologic assessment suggested a significant association between TRPV2 expression and portal vein invasion and histopathologic differentiation (P = 0.036 and 0.001, respectively). Our data suggest that TRPV2 plays a role in human hepatocarcinogenesis and might be a prognostic marker of patients with HCC.

High expression of vanilloid receptor-1 is associated with better prognosis of patients with hepatocellular carcinoma.

The vanilloid receptor-1 (VR1) is a ligand-gated, nonselective cation channel expressed predominantly by sensory neurons, but is also involved in carcinogenesis. To elucidate its role in hepatocarcinogenesis, we analyzed the expression of VR1 receptor in tumor and nontumor tissues from human hepatocellular carcinoma (HCC) samples. In situ hybridization analysis showed overexpression of VR1 mRNAs in 9/15 (60.0%) noncancer and 6/15 (40.0%) HCC samples. Immunohistochemistry of 62 HCC samples showed the expression of VR1 increased from normal liver or chronic hepatitis to cirrhosis. Marked expression of VR1 was noted in the majority [31/38 (81.6%)] of cirrhotic liver samples. In HCC, high expression of VR1 was observed in 30/62 (48.4%) cases. Clinicopathologic evaluation indicated a significant correlation between VR1 expression and histopathologic differentiation (P=0.001). Univariate analysis indicated that disease-free survival was significantly better in HCC patients with high versus those with low VR1 expression levels (P= 0.021). Our results indicate that VR1 has anti-HCC progression effects and can be potentially used as a prognostic indicator of HCC. The results suggest the potential beneficiary effects of VR1 expression on the prognosis of patients with HCC.