Metagenomic Sequencing From Mosquitoes in China Reveals a Variety of Insect and Human Viruses

We collected 8,700 mosquitoes in three sites in China, which belonged to seven species. Their viromes were tested using metagenomic sequencing and bioinformatic analysis. The abundant viral sequences were detected and annotated belonging to more than 50 viral taxonomic families. The results were verified by PCR, followed by phylogenetic analysis. In the present study, we identified partial viral genes of dengue virus (DENV), a novel circovirus (CCV), densovirus (DNV), Japanese encephalitis virus (JEV), and Wuhan mosquito virus (WMV) in mosquitoes. Metagenomic analysis and PCR amplification revealed three DENV sequences, which were as homologous to the NS3 gene of DENV from Singapore isolated in 2005, with at least 91% nucleotide (nt) identity. Seven fragments of JEV encoding structural proteins were identified belonging to genotype I. They all shared high homology with structural protein genes of JEV isolated from Laos in 2009. The production of infectious virus particles of the newly isolated virus YunnanJEV2017-4 increased after passage from the BHK-21 cell line to the Vero cell line. Novel circovirus-related genes were identified and as being related to an unnamed gene of a mosquito circovirus (MCCV) sequence from the USA isolated in 2011, with at least 41% nt identity: this distant relationship suggests that the parent virus might belong to a novel circovirus genus. Additionally, numerous known viruses and some unknown viruses were also detected in mosquitoes from Yunnan province, China, which will be tested for propagation

Metagenomic Analysis of Flaviviridae in Mosquito Viromes Isolated From Yunnan Province in China Reveals Genes From Dengue and Zika Viruses

More than 6,000 mosquitoes of six species from six sites were collected and tested for their virome using metagenomics sequencing and bioinformatic analysis. The identified viral sequences belonged to more than 50 viral families. The results were verified by PCR of selected viruses in all mosquitoes, followed by phylogenetic analysis. In the present study, we identified the partial dengue virus (DENV), Zika virus (ZIKV), and Japanese encephalitis virus (JEV) sequences in mosquitoes. Metagenomic analysis and the PCR amplification revealed three DENV sequences, one of which encodes a partial envelope protein. Two ZIKV sequences both encoding partial nonstructural protein 3 and one JEV sequence encoding the complete envelope protein were identified. There was variability in the viral titers of the newly isolated virus JEV-China/YN2016-1 of different passage viruses. The newly identified Zika virus gene from ZIKV-China/YN2016-1 was an Asian genotype and shared the highest nucleotide sequence identity (97.1%) with a ZIKV sequence from Thailand isolated in 2004. Phylogenetic analysis of ZIKV-China/YN2016-1 and ZIKV-China/YN2016-2 with known Flavivirus genes indicated that ZIKV has propagated in Yunnan province, China

Germline Predisposition and Copy Number Alteration in Pre-stage Lung Adenocarcinomas Presenting as Ground-Glass Nodules

Objective: Synchronous multiple ground-glass nodules (SM-GGNs) are a distinct entity of lung cancer which has been emerging increasingly in recent years in China. The oncogenesis molecular mechanisms of SM-GGNs remain elusive.Methods: We investigated single nucleotide variations (SNV), insertions and deletions (INDEL), somatic copy number variations (CNV), and germline mutations of 69 SM-GGN samples collected from 31 patients, using target sequencing (TRS) and whole exome sequencing (WES).Results: In the entire cohort, many known driver mutations were found, including EGFR (21.7%), BRAF (14.5%), and KRAS (6%). However, only one out of the 31 patients had the same somatic missense or truncated events within SM-GGNs, indicating the independent origins for almost all of these SM-GGNs. Many germline mutations with a low frequency in the Chinese population, and genes harboring both germline and somatic variations, were discovered in these pre-stage GGNs. These GGNs also bore large segments of copy number gains and/or losses. The CNV segment number tended to be positively correlated with the germline mutations (r = 0.57). The CNV sizes were correlated with the somatic mutations (r = 0.55). A moderate correlation (r = 0.54) was also shown between the somatic and germline mutations.Conclusion: Our data suggests that the precancerous unstable CNVs with potentially predisposing genetic backgrounds may foster the onset of driver mutations and the development of independent SM-GGNs during the local stimulation of mutagens

Roof Hydraulic Fracturing for Preventing Floor Water Inrush under Multi Aquifers and Mining Disturbance: A Case Study

Water inrush disasters from the coal seam floor occur frequently due to the high water pressure of the Ordovician limestone aquifer, multiple aquifers and strong mining disturbance. We presented a model of water-resisting key strata (WRKS) to investigate the mechanism of floor water inrush from multiple aquifers in deep coal mines. Roof hydraulic fracturing (RHF) for controlling floor water inrush and multi-parameter monitoring were proposed and validated in the Xingdong coal mine in Xingtai, Hebei Province. The results indicated that the periodic weighting step of the test working face after RHF was 9.53 m, which was 61.42% less than that of the working face without RHF (24.7 m). The floor failure depth was 30 m, which was 34.4% less than that of the zones without RHF (45.7 m). Hydraulic fracturing weakened the strength of the overlying strata to control the weighting step and reduce the mining disturbance stress, and the stability of the floor WRKS was enhanced, thereby preventing water inrush from the coal seam floor. The research results provide a solution for preventing floor damage and floor water inrush under strong mining disturbance and in complex hydrogeological environments in deep mining

Roof Hydraulic Fracturing for Preventing Floor Water Inrush under Multi Aquifers and Mining Disturbance: A Case Study

Water inrush disasters from the coal seam floor occur frequently due to the high water pressure of the Ordovician limestone aquifer, multiple aquifers and strong mining disturbance. We presented a model of water-resisting key strata (WRKS) to investigate the mechanism of floor water inrush from multiple aquifers in deep coal mines. Roof hydraulic fracturing (RHF) for controlling floor water inrush and multi-parameter monitoring were proposed and validated in the Xingdong coal mine in Xingtai, Hebei Province. The results indicated that the periodic weighting step of the test working face after RHF was 9.53 m, which was 61.42% less than that of the working face without RHF (24.7 m). The floor failure depth was 30 m, which was 34.4% less than that of the zones without RHF (45.7 m). Hydraulic fracturing weakened the strength of the overlying strata to control the weighting step and reduce the mining disturbance stress, and the stability of the floor WRKS was enhanced, thereby preventing water inrush from the coal seam floor. The research results provide a solution for preventing floor damage and floor water inrush under strong mining disturbance and in complex hydrogeological environments in deep mining

The Changes in Soil Microorganisms and Soil Chemical Properties Affect the Heterogeneity and Stability of Soil Aggregates before and after Grassland Conversion

The conversion of grasslands to croplands is common in the agro-pastoral ecotone and brings potential risks to soil health and environmental safety. As the forming unit of soil structure, the status of soil aggregates determines soil health and is affected by multiple factors. This study investigated the changes in soil aggregate and main related factors in conversion grasslands with different managed years. Grassland conversion ages were selected as experimental treatments, which included unmanaged grassland, 3 years, 10 years, 30 years, and 50 years since grassland conversion. After grassland conversion, the proportion of large macro-aggregates with a particle size of >2 mm in the 0–10 cm soil layer decreased, small macro-aggregates with a particle size of 2–0.25 mm and micro-aggregates with a particle size of 0.25–0.053 mm increased, while aggregates with a particle size of <0.053 mm had no significant change. Soil chemical properties, most microorganisms and the soil aggregate stability indices MWD and GMD decreased at the early stage (<30 years) of the managed grasslands. After about 50 years of cultivation, soil chemical properties and microorganisms returned to equal or higher levels compared to unmanaged grasslands. However, the stability of aggregates (mean weight diameter (MWD) and geometric mean diameter (GMD)) did not recover to the initial state. MWD and GMD were positively correlated with most bacterial factors (total phospholipid fatty acids (PLFAs), bacteria, Gram-positive bacteria, Gram-negative bacteria, actinomycetes and arbuscular mycorrhizal fungi (AMF)) and some soil chemical properties (carbon, nitrogen and polysaccharides). According to the partial least square structural equation model, soil organic carbon, total nitrogen and phosphorus in the 0–10 cm soil layer explained 33.0% of the variance in MWD by influencing microorganisms. These results indicated that the stability of aggregates was directly driven by microorganisms and indirectly affected by soil organic carbon, total nitrogen and phosphorus

Isolation and Characterization of Poecistasin, an Anti-Thrombotic Antistasin-Type Serine Protease Inhibitor from Leech Poecilobdella manillensis

Antistasin, first identified as a potent inhibitor of the blood coagulation factor Xa, is a novel family of serine protease inhibitors. In this study, we purified a novel antistasin-type inhibitor from leech Poecilobdella manillensis called poecistasin. Amino acid sequencing of this 48-amino-acid protein revealed that poecistasin was an antistasin-type inhibitor known to consist of only one domain. Poecistasin inhibited factor XIIa, kallikrein, trypsin, and elastase, but had no inhibitory effect on factor Xa and thrombin. Poecistasin showed anticoagulant activities. It prolonged the activated partial thromboplastin time and inhibited FeCl3-induced carotid artery thrombus formation, implying its potent function in helping Poecilobdella manillensis to take a blood meal from the host by inhibiting coagulation. Poecistasin also suppressed ischemic stroke symptoms in transient middle cerebral artery occlusion mice model. Our results suggest that poecistasin from the leech Poecilobdella manillensis plays a crucial role in blood-sucking and may be an excellent candidate for the development of clinical anti-thrombosis and anti-ischemic stroke medicines

A New Parameter Identification Method for Industrial Robots with Friction

Commonly used intelligent algorithms that are used to identify the parameters of friction of industrial robots have poor accuracy or involve complex coding, which is not conducive to their use in engineering. This paper uses the random wandering simulated annealing-based variable-step beetle antennae search (RWSAVSBAS) algorithm to identify the parameters of friction of industrial robots. The moment of friction of the third joint of the robot is experimentally obtained and used to establish a Stribeck friction model. Following this, the RWSAVSBAS algorithm is used to identify the frictional parameters of the industrial robot. These parameters can be used to accurately predict the friction-induced torque of the robot

A New Parameter Identification Method for Industrial Robots with Friction

Commonly used intelligent algorithms that are used to identify the parameters of friction of industrial robots have poor accuracy or involve complex coding, which is not conducive to their use in engineering. This paper uses the random wandering simulated annealing-based variable-step beetle antennae search (RWSAVSBAS) algorithm to identify the parameters of friction of industrial robots. The moment of friction of the third joint of the robot is experimentally obtained and used to establish a Stribeck friction model. Following this, the RWSAVSBAS algorithm is used to identify the frictional parameters of the industrial robot. These parameters can be used to accurately predict the friction-induced torque of the robot