Assembly processes and co-occurrence of bacterial communities in tree rhizosphere under Pb-Zn contamination.

Rhizosphere bacteria are critical for supporting plant performance in stressful environments. Understanding the assembly and co-occurrence of rhizosphere bacterial communities contributes significantly to both plant growth and heavy metal accumulation. In this study, Ligustrum lucidum and Melia azedarach were planted in soils with simulated varying levels of Pb-Zn contamination. The Rhizosphere bacterial communities were investigated by using 16S rRNA gene sequencing. The impacts of Pb-Zn contamination on the diversity and structure of the rhizosphere bacterial community were found to be greater than those of both tree species. The variation in bacterial community structure in both trees was mainly driven by the combinations of Pb-Zn and soil properties. Deterministic processes (non-planted, 82 %; L. lucidum, 73 %; M. azedarach, 55 %) proved to be the most important assembly processes for soil bacterial communities, but both trees increased the importance of stochastic processes (18 %, 27 %, 45 %). The rhizosphere co-occurrence networks exhibited greater stability compared to the non-planted soil networks. Rare taxa played a dominant role in maintaining the stability of rhizosphere networks, as most of the keystone taxa within rhizosphere networks belonged to rare taxa. Dissimilarities in the structure and network complexity of rhizosphere bacterial communities were significantly associated with differences in tree biomass and metal accumulation. These variations in response varied between both trees, with L. lucidum exhibiting greater potential for phytoremediation in its rhizosphere compared to M. azedarach. Our results offer valuable insights for designing effective microbe-assisted phytoremediation systems.

Regulatory network analysis based on integrated miRNA-TF reveals key genes in heart failure.

The etiology and pathophysiology of heart failure are still unknown. Increasing evidence suggests that abnormal microRNAs (miRNAs) and transcription factors (TFs) expression may be associated with the development of heart failure. Therefore, this study aims to explore key miRNAs, TFs, and related genes in heart failure to gain a greater understanding of the pathogenesis of heart failure. To search and download the dataset of mRNA chips related to heart failure from the GEO database (GSE59867, GSE9128, and GSE134766), we analyzed differential genes and screened the common differentially expressed genes on two chips using R language software. The binary interactions and circuits among miRNAs, TFs, and corresponding genes were determined by Pearson correlation coefficient. A regulatory network of miRNAs, TFs, and target genes was constructed based on bioinformatics. By comparing the sequences of patients with and without heart failure, five downregulated genes with hypermethylated mRNA and three upregulated genes with hypomethylated mRNA were identified. The miRNA-TF gene regulatory network consisted of 26 miRNAs, 22 TFs and six genes. GO and KEGG analysis results revealed that BP terms like cellular response to organic substance, cellular response to cytokine stimulus, and KEGG pathways like osteoclast differentiation, MAPK signaling pathway, and legionellosis were enriched of the DEGs. TMEM87A, PPP2R2A, DUSP1, and miR-92a have great potential as biomarkers for heart failure. The integrated analysis of the mRNA expression spectrum and microRNA-transcription factor-gene revealed the regulatory network of heart failure, which may provide clues to its alternative treatment.

Phenolic compounds weaken the impact of drought on soil enzyme activity in global wetlands.

Soil enzymes play a central role in carbon and nutrient cycling, and their activities can be affected by drought-induced oxygen exposure. However, a systematic global estimate of enzyme sensitivity to drought in wetlands is still lacking. Through a meta-analysis of 55 studies comprising 761 paired observations, this study found that phosphorus-related enzyme activity increased by 38% as result of drought in wetlands, while the majority of other soil enzyme activities remained stable. The expansion of vascular plants under long-term drought significantly promoted the accumulation of phenolic compounds. Using a 2-week incubation experiment with phenol supplementation, we found that phosphorus-related enzyme could tolerate higher biotoxicity of phenolic compounds than other enzymes. Moreover, a long-term (35 years) drainage experiment in a northern peatland in China confirmed that the increased phenolic concentration in surface layer resulting from a shift in vegetation composition inhibited the increase in enzyme activities caused by rising oxygen availability, except for phosphorus-related enzyme. Overall, these results demonstrate the complex and resilient nature of wetland ecosystems, with soil enzymes showing a high degree of adaptation to drought conditions. These new insights could help evaluate the impact of drought on future wetland ecosystem services and provide a theoretical foundation for the remediation of degraded wetlands.

Spatial Response of Ecosystem Service Value to Urbanization in Fragile Vegetation Areas Based on Terrain Gradient.

The contradiction between urban expansion and ecological protection in fragile vegetation areas has become increasingly prominent with regional development. Revealing the relationship between urbanization and ecosystem services will help to provide solutions to this problem. In order to clarify the impact of urbanization on typical mountain areas with fragile vegetation on the Qinghai Tibet Plateau, we built an ecosystem service value (ESV) evaluation index system. We also evaluated the ESV and its spatial response to the urbanization of Shannan Prefecture in Tibet from 1990 to 2015 based on different terrain gradients (TGs) using vegetation biophysical data obtained from remote sensing platforms. The results show that ESV in Shannan increased first and then declined as the TG increased, reaching a maximum value at the third TG. ESV showed a decreased trend during the study period, with a significant decline at the second and third TGs, which were the main distribution areas of vegetation in Shannan. Through spatial correlation analysis, we observed that urbanization and ESV showed a significant spatial aggregation effect. Among them, the high-low type accounted for the largest proportion in the grid with the agglomeration effect, mainly concentrated at the lower TG in the southern of Shannan, where ESV decreases with the increasing urbanization. We highlight the need for targeted, sustainable development policies to rationally organize the urbanization process in the different-gradient plateau regions with fragile vegetation. These results can provide a reference for applying ESV to vegetation restoration and ecological protection in ecologically fragile mountain areas.

Geographical and Ecological Differences in Pepper Cultivation and Consumption in China.

Peppers (Capsicum spp.) are used as food items, and particularly condiments, across most of the world. Accordingly, these vegetables occupy the largest annual stable planting area (>21,000 km2) in China. However, pepper growth, cultivation systems, yield formation, and cultivar traits vary among different environments. China is characteristic for its widely diverse terrains and high ecological heterogeneity, which determine its unique pepper consumption habits and cultivation patterns. The present study provides a comprehensive overview and analysis of the geographical and ecological characteristics of Chinese pepper consumption habits and cultivation systems, and the influence of climatic and human factors on the national pepper planting industry. For this, we analyzed detailed geospatial datasets and reviewed relevant policy papers and academic literature. Based on those findings, we then proposed sustainable management strategies for China's pepper industry; we offered suggestions for aligning the continued development of pepper cultivation with the national objective of achieving an ecological civilization and the nutritional requirements of an increasingly affluent and diverse population.

Using gross ecosystem product (GEP) to value nature in decision making.

Gross domestic product (GDP) summarizes a vast amount of economic information in a single monetary metric that is widely used by decision makers around the world. However, GDP fails to capture fully the contributions of nature to economic activity and human well-being. To address this critical omission, we develop a measure of gross ecosystem product (GEP) that summarizes the value of ecosystem services in a single monetary metric. We illustrate the measurement of GEP through an application to the Chinese province of Qinghai, showing that the approach is tractable using available data. Known as the "water tower of Asia," Qinghai is the source of the Mekong, Yangtze, and Yellow Rivers, and indeed, we find that water-related ecosystem services make up nearly two-thirds of the value of GEP for Qinghai. Importantly most of these benefits accrue downstream. In Qinghai, GEP was greater than GDP in 2000 and three-fourths as large as GDP in 2015 as its market economy grew. Large-scale investment in restoration resulted in improvements in the flows of ecosystem services measured in GEP (127.5%) over this period. Going forward, China is using GEP in decision making in multiple ways, as part of a transformation to inclusive, green growth. This includes investing in conservation of ecosystem assets to secure provision of ecosystem services through transregional compensation payments.

Morphology and Molecular Analysis of Moesziomyces antarcticus Isolated From the Blood Samples of a Chinese Patient.

Objective: To identify the pathogen causing fungemia in a Chinese patient and describe its morphological and molecular characterizes. Methods: Samples of central and peripheral venous blood were collected for blood culture. Morphology and drug sensitivities of the isolated yeast-like fungus were analyzed. rDNA sequencing and molecular phylogenetic analysis of the isolated strains were performed using DNAMAN and MEGA software. Results: A strain of yeast-like fungi was repeatedly isolated from blood samples of a Chinese patient. The isolates grew well on sabouraud medium broth plate. The colonies were smooth and round at 28°C, and were of rough surface and irregular shape at 35°C. Molecular phylogenetic trees constructed based on the internal transcribed spacer (ITS) and D1/D2 domains of 28S rDNA gene demonstrated the isolated yeast-like fungus was Moesziomyces antarcticus. Drug susceptibility test showed that this isolated M. antarcticus was resistant or had relatively low susceptibility to flucytosine, fluconazole, voriconazole, and itraconazole, and only sensitive to amphotericin. Conclusion: This study provided more information for the molecular and morphology characteristics of M. antarcticus and reviewed the species information of Moesziomyces associated with human infections, which will contribute to the identification and diagnosis of Moesziomyces infections.

Hepatitis C Virus Entry into Macrophages/Monocytes Mainly Depends on the Phagocytosis of Macrophages.

BACKGROUND: Hepatitis C virus (HCV) has been classified as a strictly hepatotropic pathogen for a long time, and hepatocytes are target cells for HCV infection. More and more studies showed non-liver cells supported HCV entry and replication, such as macrophages. The mechanisms of HCV entry into macrophages are still not clear. AIMS: This study aims to determine the way of HCV entry into macrophages. METHODS: Cell culture-derived infectious HCV particles (HCVcc) were prepared using Huh7 cells transfected with HCV RNA. CD81-knockdown cells were obtained through siRNA transfection. HCV RNA levels were determined by RT-qPCR. Flow cytometry analyses were used to determine cell surface levels of CD11b, CD68, and CD81. ELISA and western blotting were performed to quantify the protein levels of IL-1ß, IL-6, and TNF-α. Phagocytic ability was determined by neutral red uptake assay. RESULTS: CD81 knockdown could not inhibit HCVcc entry into macrophages. The entry of HCV into macrophages could not be blocked by pooled IgG from chronic hepatitis C patient's sera. Macrophages derived from THP-1 cells displayed stronger phagocytic capacity, which also swallowed more HCV RNA. Treatment of macrophages with endocytic inhibitor, methyl-ß-cyclodextrin, decreased the internalization of HCV. HCV uptake by macrophages was related to the reorganization of F-actin cytoskeleton and PI3Ks activation. HCV infection significantly increased the expression of IL1ß and IL6 in macrophages and promoted apoptosis of macrophages. CONCLUSIONS: HCV entry into macrophages mainly depends on phagocytosis of macrophages.

Elevated MIC Values of Imidazole Drugs against Aspergillus fumigatus Isolates with TR34/L98H/S297T/F495I Mutation.

The use of azole fungicides in agriculture is believed to be one of the main reasons for the emergence of azole resistance in Aspergillus fumigatus Though widely used in agriculture, imidazole fungicides have not been linked to resistance in A. fumigatus This study showed that elevated MIC values of imidazole drugs were observed against A. fumigatus isolates with TR34/L98H/S297T/F495I mutation, but not among isolates with TR34/L98H mutation. Short-tandem-repeat (STR) typing analysis of 580 A. fumigatus isolates from 20 countries suggested that the majority of TR34/L98H/S297T/F495I strains from China were genetically different from the predominant major clade comprising most of the azole-resistant strains and the strains with the same mutation from the Netherlands and Denmark. Alignments of sterol 14α-demethylase sequences suggested that F495I in A. fumigatus was orthologous to F506I in Penicillium digitatum and F489L in Pyrenophora teres, which have been reported to be associated with imidazole resistance. In vitro antifungal susceptibility testing of different recombinants with cyp51A mutations further confirmed the association of the F495I mutation with imidazole resistance. In conclusion, this study suggested that environmental use of imidazole fungicides might confer selection pressure for the emergence of azole resistance in A. fumigatus.

Whole-genome analyses of human adenovirus type 55 emerged in Tibet, Sichuan and Yunnan in China, in 2016.

Three outbreaks of acute respiratory disease occurred at military camps in 2016 at Tibet, Sichuan and Yunnan province, China. The pathogen induced these three outbreaks were all confirmed as HAdV-55 by genotype-specific PCR. The outbreak in Tibet was the first report that HAdV-55 occurred in the high altitude (HA, above sea level 3658 m). This study aims to determine the gene variation and evolution characteristics of these viral strains. Three strains of adenoviruses, LS89/Tibet/2016 (GenBank accession no. KY002683), SF04/SC/2016 (GenBank accession no. KY002684) and KM03/YN/2016 (GenBank accession no. KY002685) were obtained and confirmed by wholegenome sequencing. No multi-gene fragments recombination were found in these isolated HAdV-55 virus compared with previous reported HAdV-55 strains in China. The outbreaks in Tibet and in Sichuan continuously occurred. Virus isolated from Tibet (LS89/Tibet/2016) and Sichuan (SF04/SC/2016) had a similar mutation pattern and had a closer genetic evolutionary distance than KM03/YN/2016 strain, which indicates that the pathogens causing these two outbreaks may be of the same origin. Moreover, we found that heating was an effective way to inactive these viruses, which provide valuable information for the development of HAdV-55 vaccines. Our data provide new information for genetic evolution of HAdV-55, and contribute to the prevention and control of HAdV-55 infection in the future.

Monocyte chemoattractant protein 1 released from macrophages induced by hepatitis C virus promotes monocytes migration.

Hepatitis C Virus (HCV) infection usually progress to chronic liver disease and shows a significant increase in total monocyte/macrophages numbers in the liver. Monocyte chemoattractant protein-1 (MCP-1) plays a role in the recruitment of monocytes to the liver. In this study we found that MCP-1 were up-regulated in macrophages cultured with cell-culture derived infectious HCV particles (HCVcc) and promoted the migration of monocytes. IL1ß, IL6 and TNFα were factors that induced MCP-1 expression, which were up-regulated in macrophages induced by HCV. Long-term of HCV incubation induced apoptosis of macrophages. Finally, we observed the effect of HCV infected macrophages on nearby liver cells. Huh7 cells continuously co-cultured with monocyte/macrophages displayed increased expression of pro-inflammatory cytokines and the morphology of Huh7 cells were greatly changed. Taken together, our study provides more information for the role of monocyte/macrophages in HCV related chronic liver disease.

Epidemiology and Molecular Characterizations of Azole Resistance in Clinical and Environmental Aspergillus fumigatus Isolates from China.

Azole resistance in Aspergillus fumigatus has emerged as a worldwide public health problem. We sought here to demonstrate the occurrence and characteristics of azole resistance in A. fumigatus from different parts of China. A total of 317 clinical and 144 environmental A. fumigatus isolates from 12 provinces were collected and subjected to screening for azole resistance. Antifungal susceptibility, cyp51A gene sequencing, and genotyping were carried out for all suspected azole-resistant isolates and a subset of azole-susceptible isolates. As a result, 8 (2.5%) clinical and 2 (1.4%) environmental A. fumigatus isolates were identified as azole resistant. Five azole-resistant strains exhibit the TR34/L98H mutation, whereas four carry the TR34/L98H/S297T/F495I mutation in the cyp51A gene. Genetic typing and phylogenetic analysis showed that there was a worldwide clonal expansion of the TR34/L98H isolates, while the TR34/L98H/S297T/F495I isolates from China harbored a distinct genetic background with resistant isolates from other countries. High polymorphisms existed in the cyp51A gene that produced amino acid changes among azole-susceptible A. fumigatus isolates, with N248K being the most common mutation. These data suggest that the wide distribution of azole-resistant A. fumigatus might be attributed to the environmental resistance mechanisms in China.

Hepatitis C virus infection induces elevation of CXCL10 in human brain microvascular endothelial cells.

Hepatitis C virus (HCV) primarily infects liver tissues, while pathogenesis of extrahepatic tissues has been reported. About 50% of patients with HCV infection suffer from neurological disease. The underlying molecular mechanisms remain unclear. In the present study, we aimed to investigate the induction of CXC chemokine ligand 10 (CXCL10) in human brain microvascular endothelial cells (HBMECs) by HCV infection. CXCL10 and its receptor CXCR3 were constitutively expressed in HBMECs. HCV infection induced CXCL10 elevation in HBMECs. The elevation of CXCL10 in HBMECs was eliminated when HCV infection was blocked by neutralizing antibodies. NF-κB is a positive regulator for CXCL10 transcription. HCV infection led to an increased phosphorylation of NF-κB (ser536) in HBMECs, and CXCL10 induced by HCV was slightly decreased when an inhibitor of NF-κB was added. IL1 beta and IFN gama were also upregulated in HCV infected HBMECs, and could be depressed by inhibitor of NF-κB. Thus, HCV infection leads to upregulated expression of CXCL10 in HBMECs, which is probably via the phosphorylation of NF-κB. The findings of this study provide potential mechanisms and novel targets for HCV induced neuroinflammation. J. Med. Virol. 88:1596-1603, 2016. © 2016 Wiley Periodicals, Inc.

LRRFIP1 Inhibits Hepatitis C Virus Replication by Inducing Type I Interferon in Hepatocytes.

BACKGROUND: Hepatitis C virus infection is one of the leading causes of end stage liver diseases. The innate immune response slows down viral replication by activating cytokines such as type I interferon (IFN-α/ß), which trigger the synthesis of antiviral proteins and modulate the adaptive immune system. Recently, leucine-rich repeat (in Flightless I) interacting protein-1 (LRRFIP1) was reported contributing to the production of interferon-ß in macrophages. OBJECTIVES: The aim of this study was to assess the role of LRRFIP1 in induction of IFN-ß and inhibition of HCV infection in hepatocytes. MATERIALS AND METHODS: Induction of IFN-ß by LRRFIP1 in Huh7 and Huh7.5.1 was determined by real-time PCR and western blotting in vitro. Inhibition of HCV replication by LRRFIP1 overexpression in hepatocytes was assessed. RESULTS: LRRFIP1 increased the expression of IFN-ß in hepatocytes with or without HCV infection. Induction of IFN-ß by LRRFIP1 was enhanced with the presence of hepatitis C virus. Overexpression of LRRFIP1 in hepatocytes inhibited HCV replication. However, HCV infection did not regulate intracellular expression of LRRFIP1. CONCLUSIONS: LRRFIP1 and its mediated production of type I interferon play a role in controlling HCV infection. The findings of this study provide new target for HCV treatment and contribute to development of anti-HCV drugs.

Characteristics of qacA/B-positive Staphylococcus aureus isolated from patients and a hospital environment in China.

OBJECTIVES: This study was designed to demonstrate the characteristics of qacA/B-positive Staphylococcus aureus in China. METHODS: One hundred and forty-five MRSA and 178 MSSA from clinical specimens from seven hospitals in different regions of China, 70 MRSA from superficial sites of patients and 106 MRSA from environmental samples from an ICU were collected and screened for the presence of the qacA/B gene. The qacA/B-positive isolates and 72 randomly selected qacA/B-negative control isolates were further characterized by MLST, spa typing and detection of toxin genes, as well as antimicrobial and chlorhexidine susceptibility. SCCmec typing was conducted for MRSA. PFGE was conducted for qacA/B-positive isolates. RESULTS: Twenty-five (7.8%) of the 321 MRSA isolates harboured qacA/B, including 11 isolates from clinical specimens (7.6%), 12 isolates from patients' superficial sites (17.1%) and 2 isolates from an ICU environment (1.9%). Ten and five qacA/B-positive MRSA were identified as ST239-t030-MRSA-III and ST239-t037-MRSA-III, respectively. Six PFGE clusters and five singletons were identified among the 25 qacA/B-positive MRSA. Only one (0.6%) of the 178 MSSA isolates harboured qacA/B. qacA/B carriage in MRSA was statistically associated with spa-t037 and the presence of mupA. Compared with qacA/B-negative MRSA, the qacA/B-positive MRSA exhibited a lower susceptibility to chlorhexidine and higher resistance rates to clindamycin and trimethoprim/sulfamethoxazole. CONCLUSIONS: Carriage of qacA/B, although it had a low prevalence, might be the main reason for declining susceptibility to chlorhexidine in MRSA from Chinese patients and is probably associated with spa-t037 and the presence of the mupA gene.

Analysis of telomerase activity based on a spired DNA tetrahedron TS primer.

The development of sensitive telomerase biosensors is hindered by the restricted accessibility of telomere strand (TS) primer and the limited enzyme reaction space, which is mainly confined by the vertical distance. In this work, we designed an electrochemical telomerase biosensor based on a spired DNA tetrahedron TS primer (STTS). By adding a rigid dsDNA spire onto the top of the DNA tetrahedron, we successfully regulated the distance between the TS primer and the surface, and thus greatly facilitated the telomerase elongation on surface. The signal-to-noise ratio was 2 times higher than TSP without the spire structure. The limit of detection was calculated to be lower than 10 HeLa cells, which is at least 2 magnitudes lower than other surface extension-based electrochemical telomerase sensors without amplification. The practicability of STTS sensor was also demonstrated by analysing various other cell lines including cancer cells, stem cells of high telomerase activity and somatic cells of low telomerase activity.

Direct identification of Mycobacterium abscessus through 16S rDNA sequence analysis and a citrate utilization test: A case report.

A growing number of nontuberculous mycobacteria infection cases, especially those caused by rapidly growing mycobacteria (RGM), have been reported in the past decade. Conventional methods for mycobacteria diagnosis are inefficient and easily lead to misdiagnosis. New detection methods, such as gene sequencing, have been reported but are not widely used. The aim of the present case report was to provide a quick and exact method of identifying Myobacterium abscessus (M. abscessus) infections. The particular case reported in this study initially manifested as hyperglycemia and papules in the right leg. Routine cultures for fungus were repeatedly negative. However, cultures of the purulent material under aerobic cultivation for five days yielded a rapidly growing, nontuberculous mycobacterium. A Ziehl-Neelsen staining of this mycobacterium revealed the presence of acid-fast bacilli that were finally identified as M. abscessus through 16S rDNA sequence analysis and a citrate utilization test. The current report may help other clinicians to make a quick and accurate diagnosis of RGM infection.

CXCL10 decreases GP73 expression in hepatoma cells at the early stage of hepatitis C virus (HCV) infection.

Golgi protein 73 (GP73), which is up-regulated in hepatocellular carcinoma (HCC), has recently been identified as a novel serum marker for HCC diagnosis. Several reports also noted the increased levels of GP73 expression in chronic liver disease in patients with acute hepatitis of various etiologies, chronic Hepatitis C virus (HCV) infection and alcoholic liver disease. The molecular mechanisms of GP73 expression in HCV related liver disease still need to be determined. In this study, we aimed to evaluate the effect of HCV infection on GP73 expression. GP73 was highly expressed in Huh7, Hep3B, 293T and HUVEC cells, and was low-expressed in HepG2 cells. HCV infection led to down-regulation of GP73 in Huh7 and HepG2/CD81 cells at the early stage of infection. CXCL10 decreased GP73 expression in Huh7 and HepG2 cells. Up-regulation of GP73 was noted in hepatocytes with cytopathic effect at advanced stage of HCV infection, and further research is needed to determine the unknown factors affecting GP73 expression. In conclusion, our study provided additional evidence for the roles of GP73 in liver disease.

A carbon nanotube-based high-sensitivity electrochemical immunosensor for rapid and portable detection of clenbuterol.

Carbon nanotubes have shown their unique advantages of mechanical, chemical and electronic properties in bioanalysis. We herein report a new method to efficiently and reproducibly prepare multi-walled carbon nanotubes (MWNTs)-protein sensing layers for electrochemical immunosensors. This method employs centrifugation to prepare a conjugate of MWNTs and goat anti mouse-immunoglobulin G (IgG) (secondary antibody). The conjugates were then deposited on screen-printed electrodes to form a nanostructured layer (MWNT-I layer). CLB monoclonal antibody was assembled through its binding to the secondary antibody. The MWNT-I layer-based electrodes were used for rapid and sensitive amperometric immunosensing detection of clenbuterol (CLB) in swine urine samples. Horseradish peroxidase-coupled CLB (CLB-HRP) competed with free CLB in the samples to bind the monoclonal antibody. It has shown significantly higher sensitivity and better reproducibility than the chemical conjugation method. This MWNT-based immunosensor is highly sensitive, leading to a limit of detection of 0.1 ng/mL within a rapid assay time of 16 min. Its sensitivity is at least 1 order of magnitude higher than that of a normal immunosensor (without MWNTs). The sensing device is portable with disposable screen-printed electrode, satisfactorily meeting the requirements for field detection of food security-related species.