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ABSTRACT The emergence of nosocomial infections caused by hypervirulent and carbapenem-resistant K. pneumoniae (hv-CRKP) has become a significant public health challenge. The genetic traits of virulence and resistance plasmids in hv-CRKP have been extensively studied; however, research on the adaptive evolution strategies of clinical strains inside the host was scarce. This study aimed to understand the effects of antibiotic treatment on the phenotype and genotype characteristics of hv-CRKP. We investigated the evolution of hv-CRKP strains isolated from the same patient to elucidate the transition between hospital invasion and colonization. A comparative genomics analysis was performed to identify single nucleotide polymorphisms in the rmpA promoter. Subsequent validation through RNA-seq and gene deletion confirmed that distinct rmpA promoter sequences exert control over the mucoid phenotype. Additionally, biofilm experiments, cell adhesion assays, and animal infection models were conducted to illuminate the influence of rmpA promoter diversity on virulence changes. We demonstrated that the P12T and P11T promoters of rmpA possess strong activity, which leads to the evolution of CRKP into infectious and virulent strains. Meanwhile, the specific sequence of polyT motifs in the rmpA promoter led to a decrease in the lethality of hv-CRKP and enhanced cell adhesion and colonization. To summarize, the rmpA promoter of hv-CRKP is utilized to control capsule production, thereby modifying pathogenicity to better suit the host’s ecological environment.IMPORTANCEThe prevalence of hospital-acquired illness caused by hypervirulent carbapenem-resistant Klebsiella pneumoniae (hv-CRKP) is significant, leading to prolonged antibiotic treatment. However, there are few reports on the phenotypic changes of hv-CRKP in patients undergoing antibiotic treatment. We performed a comprehensive examination of the genetic evolutionary traits of hv-CRKP obtained from the same patient and observed variations in the promoter sequences of the virulence factor rmpA. The strong activity of the promoter sequences P11T and P12T enhances the consistent production of capsule polysaccharides, resulting in an invasive strain. Conversely, weak promoter activity of P9T and P10T is advantageous for exposing pili, hence improving bacterial cell attachment ability and facilitating bacterial colonization. This finding also explains the confusion of some clinical strains carrying wild-type rmpA but exhibiting a low mucoid phenotype. This adaptive alteration facilitates the dissemination of K. pneumoniae within the hospital setting.

With the rapid development of intelligent technology, the smart heightened-containment biological laboratory (sHCBL) has moved from concept to reality. Experimental activities and laboratory construction, operation, and management will undoubtedly lead to disruptive changes. Conventional laboratories are increasingly being replaced by smart laboratories; however, the key technologies involved in this transition remain at an exploratory stage. It is necessary for HCBLs to absorb the advanced ideas of smart laboratories to guarantee the establishment of biosafety and biosecurity in a more automated way. This study examines in detail sHCBL module structures, the functions of each module, laboratory operation processes, and the advanced nature of smart laboratories. It may provide a theoretical foundation for the future transformation and smart construction of sHCBLs.

Interconnect development for the new technology node requires coordinated efforts of multiple module teams working to co-optimize patterning and metallization solutions to meet performance and yield, and reliability targets. This paper presents the results of interconnect patterning optimization to improve overlay and process window control using a novel methodology, Design-for-Inspection™ (DFI). Using design-assisted voltage contrast measurement techniques, the method enables in-line test and monitoring of process induced overlay and CD variation of back-end-of line (BEOL) features built with litho-etch-litho-etch (LELE) patterning. While only some of the features of multi-color patterning scheme are chosen to be aligned directly, other combination of metal line and via colors may have uncontrolled misalignment risking open or short failures. The paper shows how the complete metrology coverage of multi-color combinations between dual patterned via and dual patterned metal lines helps driving the improvement of overlay and process margins in 14nm technology. The optimized process margin for via opens enables higher yields and better reliability.

Since it was first identified in 1956, the H11 subvariant influenza virus has been reported worldwide. However, due to the low pathogenicity of the H11 subvariant and the absence of its widespread transmission among humans, there are only a few reports on the etiology of the H11 subvariant influenza virus. Therefore, in the present study, we isolated a strain of the H11N3 avian influenza virus (AIV) from poultry feces from the live poultry market in the southeast coastal region of China. Considering that the H11 subvariant is known to cause infections in humans and to enrich the knowledge of the H11 subvariant of the avian influenza virus, the genetics, pathogenicity, and transmissibility of the isolate were studied. The phylogenetic analysis indicated that the H11N3 isolate was of Eurasian origin and carried genes closely related to duck H7N2 and H4N6. The receptor binding analysis revealed that the H11N3 isolate only acquired a binding affinity for avian-derived receptors. In the respiratory system of mice, the isolate could directly cause infection without adaptation. In addition, the results from transmission experiments and antibody detection in guinea pigs demonstrated that H11N3 influenza viruses can efficiently transmit through the respiratory tract in mammalian models. Direct infection of the H11N3 influenza virus without adaptation in the mouse models and aerosol transmission between guinea pig models confirms its pandemic potential in mammals, underscoring the importance of monitoring rare influenza virus subtypes in future studies.

During the period from 2005 to 2015 under the rule of Hu Jintao and Xi Jinping, China has achieved enormous success not only in the field of economy but also in the political and diplomatic areas. With the Silk Road Economic Belt’s initiative launched by the government of Xi in 2013, China was seeking to find alternatives for different affairs such as the South China Sea, the nuclear issue of North Korea, and Taiwan issues. The emergency of the “Strategic Breakthrough” whose aim is to stabilize the surrounding atmosphere makes it a pressing task for the Chinese diplomacy focused on Eurasia to overhaul the international order.

Crowdsourcing design for complex products has open objectives and requires collaboration among different fields. It is difficult to use a single approach to describe the process in which the number and interaction of tasks and steps of tasks are changed dynamically depending on the execution state. Motivated by modeling the collaborative business process of crowdsourcing design in complex products, a crowdsourcing hierarchical process modeling approach based on an object-oriented Petri net is proposed. From the perspective of organization and task, the crowdsourcing object interaction and design process are modeled separately. First, an organization layer modeling approach based on expert review is proposed to describe the information transfer relationship among crowdsourcing objects from the organization perspective. Then, the task layer modeling approach based on the component decomposition idea is proposed. From the perspective of the task, the task is decomposed by using a design structure matrix (DSM) and a product structure tree to model the interaction of the design process. The object-oriented Petri net-is used to realize the representation of the model. Finally, the method is verified by taking the model of the crowdsourcing design of an air cooler as an example. The model subnet is analyzed by means of an overlay tree and incidence matrix, and the reliability and robustness of the model are proven. This method provides a reference for the process modeling of the crowdsourcing design of complex products.

Since December 2019, coronavirus disease (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become a great challenge to the world's public health system. Nosocomial infections have occurred frequently in medical institutions worldwide during this pandemic. Thus, there is an urgent need to construct an effective surveillance and early warning system for pathogen exposure and infection to prevent nosocomial infections in negative-pressure wards. In this study, visualization and construction of an infection risk assessment of SARS-CoV-2 through aerosol and surface transmission in a negative-pressure ward were performed to describe the distribution regularity and infection risk of SARS-CoV-2, the critical factors of infection, the air changes per hour (ACHs) and the viral variation that affect infection risk. The SARS-CoV-2 distribution data from this model were verified by field test data from the Wuhan Huoshenshan Hospital ICU ward. ACHs have a great impact on the infection risk from airborne exposure, while they have little effect on the infection risk from surface exposure. The variant strains demonstrated significantly increased viral loads and risks of infection. The level of protection for nurses and surgeons should be increased when treating patients infected with variant strains, and new disinfection methods, electrostatic adsorption and other air purification methods should be used in all human environments. The results of this study may provide a theoretical reference and technical support for reducing the occurrence of nosocomial infections.

Abstract Background Influenza A virus (IAV) continues to pose serious threats to public health. The current prophylaxis and therapeutic interventions for IAV requires frequent changes due to the continuous antigenic drift and antigenic shift of IAV. Emerging evidence indicates that the host microRNAs (miRNAs) play critical roles in intricate host-pathogen interaction networks. Cellular miRNAs may directly target virus to inhibit its infection and be developed as potential anti-virus drugs. Methods In this study, we established a broad-spectrum anti-IAV miRNA screening method using miRanda software. The screened miRNAs were further verified by luciferase assay, viral protein expression assay and virus replication assay. Results Five cellular miRNAs (miR-188-3p, miR-345-5p, miR-3183, miR-15-3p and miR-769-3p), targeting 99.96, 95.31, 92.9, 94.58 and 97.24% of human IAV strains recorded in NCBI, respectively, were chosen for further experimental verification. Finally, we found that miR-188-3p downregulated PB2 expression at both mRNA and protein levels by directly targeted the predicted sites on PB2 and effectively inhibited the replication of IAV (H1N1, H5N6 and H7N9) in A549 cells. Conclusions This is the first report screening cellular miRNAs that broad-spectrum inhibiting IAV infection. These findings suggested that cellular miR-188-3p could be used for RNAi-mediated anti-IAV therapeutic strategies.

NbN, NbN-Ag and NbN/NbN-Ag multilayer nanocomposite films were successfully deposited by an arc ion plating system (AIP), and their microstructures, mechanical and tribological properties were systematically investigated. The results show that all the films had a polycrystalline structure, and the Ag in the Ag-doped films existed independently as a face-centered cubic phase. The content of Ag in NbN-Ag and NbN/NbN-Ag films was 20.11 and 9.07 at.%, respectively. NbN films fabricated by AIP technique had excellent mechanical properties, and their hardness and critical load were up to 44 GPa and 34.6 N, respectively. The introduction of Ag into NbN films obviously reduced the friction coefficient at room temperature, while the mechanical properties and wear resistance were degraded sharply in comparison with that of NbN films. However, the NbN/NbN-Ag films presented better hardness, H/E*, H3/E*2, adhesive strength and wear resistance than NbN-Ag films. Additionally, analysis of wear surfaces of the studied films and Al2O3 balls using 3D images, depth profiles, energy dispersive spectrometry (EDS) and Raman spectra indicated that the main wear mechanisms of NbN and NbN/NbN-Ag films were adhesive and oxidation wear with slight abrasive wear, while the severe abrasive and oxidation wear were the dominant wear mechanism for NbN-Ag films.

Environmental transmission of viruses to humans has become an early warning for potential epidemic outbreaks, such as SARS-CoV-2 and influenza virus outbreaks. Recently, an H7N9 virus, A/environment/Hebei/621/2019 (H7N9), was isolated by environmental swabs from a live poultry market in Hebei, China. We found that this isolate could be transmitted by direct contact and aerosol in mammals. More importantly, after 5 passages in mice, the virus acquired two adaptive mutations, PB1-H115Q and B2-E627K, exhibiting increased virulence and aerosol transmissibility. These results suggest that this H7N9 virus might potentially be transmitted between humans through environmental or airborne routes.

Metastasis-associated drug resistance accounts for high mortality in ovarian cancer and remains to be a major barrier for effective treatment. In this study, SKOV3/T4, a metastatic subpopulation of ovarian cancer SKOV3 cells, was enriched to explore potential interventions against metastatic-associated drug resistance. Quantitative genomic and functional analyses were performed and found that slug was significantly increased in the SKOV3/T4 subpopulation and contributed to the high resistance of SKOV3/T4. Further studies showed that slug activated c-Met in a ligand-independent manner due to elevated levels of fibronectin and provoked integrin α V function, which was confirmed by the significant correlation of slug and p-Met levels in 121 ovarian cancer patient samples. Intriguingly, c-Met inhibitor(s) exhibited greatly enhanced anti-cancer effects in slug-positive ovarian cancer models both in vitro and in vivo. Additionally, IHC analyses revealed that slug levels were highly correlated with reduced survival of ovarian cancer patients. Taken together, this study not only uncovers the critical roles of slug in drug resistance in ovarian cancer but also highlights a promising therapeutic strategy by targeting the noncanonical activation of c-Met in slug-positive ovarian cancer patients with poor prognosis. KEY WORDS: Slug, c-Met, Drug resistance, Ovarian cancer, XL184

Although many high-risk mucosal and cutaneous human papillomaviruses (HPVs) theoretically have the potential to synthesize L1 isoforms differing in length, previous seroepidemiological studies only focused on the short L1 variants, co-assembling with L2 to infectious virions. Using the multimammate mouse Mastomys coucha as preclinical model, this is the first study demonstrating seroconversion against different L1 isoforms during the natural course of papillomavirus infection. Intriguingly, positivity with the cutaneous MnPV was accompanied by a strong seroresponse against a longer L1 isoform, but to our surprise, the raised antibodies were non-neutralizing. Only after a delay of around 4 months, protecting antibodies against the short L1 appeared, enabling the virus to successfully establish an infection. This argues for a novel humoral immune escape mechanism that may also have important implications on the interpretation of epidemiological data in terms of seropositivity and protection of PV infections in general.

In this paper, the dynamic event-triggered tracking control issue is studied for a class of unknown stochastic nonlinear systems with strict-feedback form. At first, neural networks (NNs) are used to approximate the unknown nonlinear functions. Then, a dynamic event-triggered controller (DETC) is designed through the adaptive backstepping method. Especially, the triggered threshold is dynamically adjusted. Compared with its corresponding static event-triggered mechanism (SETM), the dynamic event-triggered mechanism (DETM) can generate a larger execution interval and further save resources. Moreover, it is verified by two simulation examples that show that the closed-loop stochastic system signals are ultimately fourth moment semi-globally uniformly bounded (SGUUB).

Objective We systematically investigated the cytotoxic effects of ricin in primary pulmonary alveolar macrophages (PAMs). Methods Primary PAMs were isolated from BALB/c mice. The cytotoxic effects of ricin were investigated in vitro by optical and transmission electron microscopy, detection of the inflammatory cytokine response, proteomic analysis, and subsequent biological functional analysis. Results Ricin induced shrinkage, apoptosis, vacuolization, and multi-organelle lesions in primary PAMs as demonstrated by optical and transmission electron microscopy. Ricin also induced a pronounced pro-inflammatory cytokine response in primary PAMs, including induction of tumor necrosis factor-α, interferon-γ, interleukin (IL)-1, IL-2, IL-6, IL-12, C-C motif chemokine ligand 2, and C-X-C motif chemokine ligand 2, while the anti-inflammatory cytokines IL-4 and IL-10 were less affected. Proteomic analysis and subsequent biological functional analysis identified eight proteins that were up/downregulated by ricin treatment and which might thus contribute to ricin toxicity. These proteins were involved in various functions, including redox, molecular chaperone, glycolysis, protein translation, and protein degradation functions. Conclusion The results of the present study further our understanding of the pathogenic mechanism of inhalational ricin poisoning.

Objective This study was performed to develop a murine aerosol infection model of brucellosis to investigate the pathogenicity and immune reactions induced by aerosolized Brucella and to identify key proteins associated with Brucella infection in lung tissue. Methods BALB/c mice were exposed to aerosolized Brucella melitensis 5 (M5) for 30 minutes and killed at 1, 3, 7, and 15 days post-exposure. Clinical observation, pathological analysis of lung tissue, and cytokine expression detection were then performed. Proteomic analysis based on two-dimensional electrophoresis and mass spectrometry was used to identify proteins exhibiting significant changes in expression in lung tissues during Brucella infection. Results Pathological analysis revealed alveolar wall thickening, telangiectasia with hyperemia, inflammatory cell infiltration, large areas of congestion and bleeding, and areas of focal necrosis. The T-helper 1 type immune response played an important role during aerosol infection, and 12 differentially expressed proteins were involved in the infectious process in lung tissue. Conclusion These results contribute to our understanding of the pathogenic process of Brucella in the lung tissue of BALB/c mice challenged with aerosolized Brucella . Some of the identified proteins may be potential targets in future therapeutic strategies.

Objective To develop a bioluminescence-labelled bacterial infection model to monitor the colonization and clearance process of Escherichia coli O157:H7 in the lungs of mice following influenza A virus/Puerto Rico/8/34 (H1N1) strain (IAV/PR8) infection. Methods BALB/c mice were administered IAV/PR8 or 0.01 M phosphate-buffered saline (PBS; pH 7.4) intranasally 4 days prior to intranasal administration of 1 × 10 7 colony-forming units (CFU) of E. coli O157:H7-lux. Whole-body bioluminescent signals were monitored at 10 min, 4 h, 8 h, 12 h, 16 h and 24 h post-bacterial infection. Lung bioluminescent signals and bacterial load (CFU/g) were monitored at 4 h, 8 h, 12 h, 16 h and 24 h post-bacterial infection. Results Prior IAV/PR8 infection of mice resulted in a higher level of bacterial colonization and a lower rate of bacterial clearance from the lungs compared with mice treated with PBS. There were also consistent findings between the bioluminescence imaging and the CFU measurements in terms of identifying bacterial colonization and monitoring the clearance dynamics of E. coli O157:H7-lux in mouse lungs. Conclusion This novel bioluminescence-labelled bacterial infection model rapidly detected bacterial colonization of the lungs and monitored the clearance dynamics of E. coli O157:H7-lux following IAV/PR8 infection.

The development of non-fullerene acceptor molecules have remarkably boosted power conversion efficiency (PCE) of polymer solar cells (PSCs) due to the improved spectral coverage and reduced energy loss. An introduction of fullerene molecules into the non-fullerene acceptor-based blend may further improve the photovoltaic performance of the resultant ternary PSCs. However, the underlying mechanism is still debatable. Herein, the ternary PSCs based on PBDB-T:ITIC:PC71BM blend were fabricated and its PCE was increased to 10.2% compared to 9.2% for the binary PBDB-T:ITIC devices and 8.1% for the PBDB-T:PC71BM PSCs. Systematic investigation was carried out to disclose the effect of PC71BM on the blend morphology and charge transport behavior. It is found that the PC71BM tends to intermix with the PBDB-T donor compared to the ITIC counterpart. A small amount of PC71BM in the ternary blend is helpful for ITIC to aggregate and form efficient electron-transport pathways. Accordingly, the electron mobility is increased and the density of electron traps is decreased in the ternary blend in comparison with the PBDB-T:ITIC blend. Finally, the suppressed bimolecular recombination and enhanced charge collection lead to high PCE for the ternary solar cells.

[This corrects the article DOI: 10.1371/journal.pone.0189867.].

Five full-scale wastewater treatment plants (WWTPs) in China using typical biodegradation processes (SBR, oxidation ditch, A2/O) were selected to assess the removal of four popular artificial sweeteners (ASs). All four ASs (acesulfame (ACE), sucralose (SUC), cyclamate (CYC) and saccharin (SAC)) were detected, ranging from 0.43 to 27.34μg/L in the influent. Higher concentrations of ASs were measured in winter. ACE could be partly removed by 7.11-50.76% through biodegradation and especially through the denitrifying process. The A2/O process was the most efficient at biodegrading ASs. Adsorption (by granular activated carbon (GAC) and magnetic resin) and ultraviolet radiation-based advanced oxidation processes (UV/AOPs) were evaluated to remove ASs in laboratory-scale tests. The amounts of resin adsorbed were 3.33-18.51 times more than those of GAC except for SUC. The adsorption ability of resin decreased in the order of SAC > ACE > CYC > SUC in accordance with the pKa. Degradation of ASs followed pseudo-first-order kinetics in UV/H2O2 and UV/PDS. When applied to the secondary effluent, ASs could be degraded from 30.87 to 99.93% using UV/PDS in 30 minutes and UV/PDS was more efficient and economic.

At least 15 cases of human beings infected with H5N6 have been reported since 2014, of which at least nine were fatal. The highly pathogenic avian H5N6 influenza virus may pose a serious threat to both public health and the poultry industry. However, the molecular features promoting the adaptation of avian H5N6 influenza viruses to mammalian hosts is not well understood. Here, we sequentially passaged an avian H5N6 influenza A virus (A/Northern Shoveler/Ningxia/488-53/2015) 10 times in mice to identify the adaptive amino acid substitutions that confer enhanced virulence to H5N6 in mammals. The 1st and 10th passages of the mouse-adapted H5N6 viruses were named P1 and P10, respectively. P1 and P10 displayed higher pathogenicity in mice than their parent strain. P10 showed significantly higher replication capability in vivo and could be detected in the brains of mice, whereas P1 displayed higher replication efficiency in their lungs but was not detectable in the brain. Similar to its parent strain, P10 remained no transmissible between guinea pigs. Using genome sequencing and alignment, multiple amino acid substitutions, including PB2 E627K, PB2 T23I, PA T97I, and HA R239H, were found in the adaptation of H5N6 to mice. In summary, we identified amino acid changes that are associated with H5N6 adaptation to mice.

Ebola virus emerged in West Africa in December 2013. The high population mobility and poor public health infrastructure in this region led to the development of the largest Ebola virus disease (EVD) outbreak to date.On September 26, 2014, China dispatched a Mobile Biosafety Level-3 Laboratory (MBSL-3 Lab) and a well-trained diagnostic team to Sierra Leone to assist in EVD diagnosis using quantitative real-time PCR, which allowed the diagnosis of suspected EVD cases in less than 4 hours from the time of sample receiving. This laboratory was composed of three container vehicles equipped with advanced ventilation system, communication system, electricity and gas supply system. We strictly applied multiple safety precautions to reduce exposure risks. Personnel, materials, water and air flow management were the key elements of the biosafety measures in the MBSL-3 Lab. Air samples were regularly collected from the MBSL-3 Lab, but no evidence of Ebola virus infectious aerosols was detected. Potentially contaminated objects were also tested by collecting swabs. On one occasion, a pipette tested positive for EVD. A total of 1,635 suspected EVD cases (824 positive [50.4%]) were tested from September 28 to November 11, 2014, and no member of the diagnostic team was infected with Ebola virus or other pathogens, including Lassa fever. The specimens tested included blood (69.2%) and oral swabs (30.8%) with positivity rates of 54.2% and 41.9%, respectively. The China mobile laboratory was thus instrumental in the EVD outbreak response by providing timely and reliable diagnostics.The MBSL-3 Lab significantly contributed to establishing a suitable laboratory response capacity during the emergence of EVD in Sierra Leone.

Mango (Mangifera indica L.) fruit is rich in polyphenolic compounds. Identification of individual gallotannins and evaluation of antiproliferative activities of gallotannin-rich extracts from kernel and peel of three Chinese mango cultivars were carried out in the present study. Five gallotannins, i.e. penta-, hexa-, hepta-, octa-, and nona-O-galloyl-glucoside were isolated and identified by LC-ESI-MS/MS and NMR, and quantified by HPLC. Mango kernel was a much richer source of gallotannins than its peel, and its antiproliferative effects on MDA-MB-231 breast, HepG2 liver, and HL-60 leukemia cancer cells were reported for the first time. DPPH• scavenging and antiproliferative activities of the mango kernel and peel extracts may be partially caused by penta-O-galloyl-glucoside or its analogs with different galloyl groups since penta-O-galloyl-glucoside and gallic acid also revealed high antioxidant and antiproliferative activities. Therefore, the gallotannin-rich extracts of mango kernel and peel may serve as natural sources of anticancer agents and may have important pharmaceutical potential.

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Walking is a complex, rhythmic task performed by the locomotor system. However, natural gait rhythms can be influenced by metronomic auditory stimuli, a phenomenon of particular interest in neurological rehabilitation. In this paper, we examined the effects of aural, visual and tactile rhythmic cues on the temporal dynamics associated with human gait. Data were collected from fifteen healthy adults in two sessions. Each session consisted of five 15-minute trials. In the first trial of each session, participants walked at their preferred walking speed. In subsequent trials, participants were asked to walk to a metronomic beat, provided through visually, aurally, tactile or all three cues (simultaneously and in sync), the pace of which was set to the preferred walking speed of the first trial. Using the collected data, we extracted several parameters including: gait speed, mean stride interval, stride interval variability, scaling exponent and maximum Lyapunov exponent. The extracted parameters showed that rhythmic sensory cues affect the temporal dynamics of human gait. The auditory rhythmic cue had the greatest influence on the gait parameters, while the visual cue had no statistically significant effect on the scaling exponent. These results demonstrate that visual rhythmic cues could be considered as an alternative cueing modality in rehabilitation without concern of adversely altering the statistical persistence of walking.