Platform establishment of the Cre-loxP recombination system for genetic manipulation of the Lumpy skin disease virus.

Lumpy skin disease virus (LSDV) is a rapidly emerging pathogen in Asia, including China. Genetic manipulation of the LSDV is essential for the elucidation of the pathogenic mechanism and biological function of the LSDV-encoded protein. In this study, we established a platform for the Cre-loxP recombination system under a modified early-late H5 promoter of the VACV for quick construction of the recombinant LSDV virus. The recombinant virus, LSDV-EGFP-ΔTK, was purified and obtained using serial limited dilution and picking the single cells methods. Using the lentiviral package system, a Cre recombinase enzyme stable expression MDBK cell line was established to supply the Cre recombinase for the reporter gene excision. A genetically stable, safe TK gene-deleted LSDV (LSDV-ΔTK) was constructed using homologous recombination and the Cre-loxP system. It was purified using limited dilution in the MDBK-Cre cell line. Establishing the Cre-loxP recombination system will enable sequential deletion of the interested genes from the LSDV genome and genetic manipulation of the LSDV genome, providing technical support and a platform for developing the attenuated LSDV vaccine.

Fe3O4-Cy5.5-trastuzumab magnetic nanoparticles for magnetic resonance/near-infrared imaging targeting HER2 in breast cancer.

This study developed a probe Fe3O4-Cy5.5-trastuzumab with fluorescence and magnetic resonance imaging functions that can target breast cancer with high HER2 expression, aiming to provide a new theoretical method for the diagnosis of early breast cancer. Fe3O4-Cy5.5-trastuzumab nanoparticles were combined with Fe3O4for T2imaging and Cy5.5 for near-infrared imaging, and coupled with trastuzumab for HER2 targeting. We characterized the nanoparticles used transmission electron microscopy, hydration particle size, Zeta potential, UV and Fourier transform infrared spectroscopy, and examined its magnetism, fluorescence, and relaxation rate related properties. CCK-8 and blood biochemistry analysis evaluated the biosafety and stability of the nanoparticles, and validated the targeting ability of Fe3O4-Cy5.5 trastuzumab nanoparticles throughin vitroandin vivocell and animal experiments. Characterization results showed the successful synthesis of Fe3O4-Cy5.5-trastuzumab nanoparticles with a diameter of 93.72 ± 6.34 nm. The nanoparticles showed a T2relaxation rate 42.29 mM-1s-1, magnetic saturation strength of 27.58 emg g-1. Laser confocal and flow cytometry uptake assay showed that the nanoparticles could effectively target HER2 expressed by breast cancer cells. As indicated byin vitroandin vivostudies, Fe3O4-Cy5.5-trastuzumab were specifically taken up and effectively aggregated to tumour regions with prominent NIRF/MR imaging properties. CCK-8, blood biochemical analysis and histological results suggested Fe3O4-Cy5.5-trastuzumab that exhibited low toxicity to major organs and goodin vivobiocompatibility. The prepared Fe3O4-Cy5.5-trastuzumab exhibited excellent targeting, NIRF/MR imaging performance. It is expected to serve as a safe and effective diagnostic method that lays a theoretical basis for the effective diagnosis of early breast cancer. This study successfully prepared a kind of nanoparticles with near-infrared fluorescence imaging and T2imaging properties, which is expected to serve as a new theory and strategy for early detection of breast cancer.

Identification of the murine osteoblastic cell MC3T3-E1 as a permissive cell line in response to lumpy skin disease virus.

Lumpy skin disease virus (LSDV) is a rapidly emerging pathogen in China. Screening suitable cells for LSDV replication is vital for future research on pathogenic mechanisms and vaccine development. Previous comparative studies have identified that the rodent-derived BHK21 is a highly susceptible cell model to LSDV infection. Using western blot, indirect immune-fluorescence assay, flow cytometry, and transmission electron microscopy methods, this study is the first to identify the murine osteoblastic cell line MC3T3-E1 as a novel permissive cell model for LSDV infection. The establishment of MC3T3-E1 as a suitable infectious cell model enhances our understanding of the species range and cell types of the permissive cells and nonpermissive that support LSDV replication. It is helpful to accelerate future research on the pathogenesis, clinical application, and vaccine development of LSDV.

A nomogram based on MRI radiomics features of mesorectal fat for diagnosing T2- and T3-stage rectal cancer.

PURPOSE: To develop and validate a nomogram for the preoperative diagnosis of T2 and T3 stage rectal cancer using MRI radiomics features of mesorectal fat. METHODS: The data of 288 patients with T2 and T3 stage rectal cancer were retrospectively collected. Radiomics features were extracted from the lesion region of interest (ROI) in the MRI high-resolution T2WI, apparent diffusion coefficient (ADC), and diffusion-weighted imaging (DWI) sequences. After using ICC inter-group consistency analysis and Pearson correlation analysis to reduce dimensions, LASSO regression analysis was performed to select features and calculate Rad-score for each sequence. Then, Combined_Radscore and nomogram were constructed based on the LASSO-selected features and clinical data for each sequence. Receiver operating characteristic curve (ROC) area under the curve (AUC) was used to evaluate the performance of the Rad-score model and nomogram. Decision curve analysis (DCA) was performed to evaluate the clinical usability of the radiomics nomogram, which were combined with calibration curves to evaluate the prediction accuracy. RESULTS: The nomogram based on MRI-report T status and Combined_Radscore achieved AUCs of 0.921 and 0.889 in the training and validation cohorts, respectively. CONCLUSION: The nomogram can be stated that the radiomics nomogram based on multi-sequence MRI imaging of the mesorectal fat has excellent diagnosing performance for preoperative differentiation of T2 and T3 stage rectal cancer.

Characterization of a novel recombinant NADC30­like porcine reproductive and respiratory syndrome virus in Shanxi Province, China.

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important pathogens affecting the swine industry. In this report, a novel PRRSV strain SXht2012 was isolated from Shanxi province in China. To identify genetic characteristics of SXht2012, we conducted phylogenetic and homology analyses after sequencing its complete genome. The results revealed that SXht2012 belonged to NADC30-like strain and shared 91.3% nucleotide (nt) identity with strain NADC30. Notably, sequence alignment showed that a distinctive feature in the NSP2 region, where a 131-amino acid (aa) deletion was found in the hypervariable region (HVR). Additionally, variations were also detected in the GP5 protein, specifically in the decoy peptide, T cell peptide, and a potential glycosylation site (aa 32). Furthermore, we also found that SXht2012 was likely a recombination virus originating from NADC30-like and JXA1-like strains, and three recombination breakpoints were identified in the genome at nt positions 1516, 5280 and 6851, which correspond to the NSP2, NSP3, and NSP7 regions. Overall, these findings have significant implications for understanding the genetic variation and evolutionary dynamics of PRRSV strains.

Laboratory Experiments of Hydrocarbon Gas Flooding and Its Influencing Factors on Oil Recovery in a Low Permeability Reservoir with Medium Viscous Oil.

In view of the problems of low liquid production, a high proportion of high water cut wells, and poor development effect in the late stage of water flooding in the special sandstone reservoir of Niuquanhu "low permeability and medium viscosity crude oil", we carried out the research on hydrocarbon gas oil recovery and its influencing factors. First, the influence of different injected gas media on the physical properties of crude oil was analyzed. Second, the core displacement experiments of different gas injection media including CO2, CH4, and hydrocarbon gas were carried out by using the method of oil recovery comparison and optimization. Third, the indoor experimental study on the oil recovery of different influencing factors was carried out by using the method of controlling variables of influencing factors. Finally, the influence degree of different influencing factors on oil recovery was analyzed by a Spearman rank correlation coefficient analysis. The experimental results showed that the oil recovery of hydrocarbon gas is higher than that of CO2 and CH4, which were 57, 51, and 18% respectively. This is mainly because hydrocarbon gas is similar to the components of crude oil and is more easily dissolved in crude oil. The experimental results of influencing factors showed that the higher the content of C2-C4, the higher the oil recovery, and the content of C2-C4 will affect its dissolution with crude oil and its interaction with heavy component crude oil. The larger the permeability ratio, the lower the oil recovery, which was mainly due to the uneven distribution of injected gas in different regions. The higher the permeability, the lower the oil recovery, which was also due to the serious heterogeneity of the low permeability core of Niuquanhu; The results of Spearman rank correlation coefficient analysis based on different influencing factors and oil recovery showed that the order of influence of different factors on oil recovery was C2-C4 content > permeability ratio > permeability > back pressure > gas injection rate. In the development process of hydrocarbon gas injection, we should control the C2-C4 content, back pressure, and injection rate. The research in this study not only provides theoretical support for gas injection enhanced oil recovery technology in "low permeability and medium viscosity crude oil" reservoirs but also provides a new idea for the ranking of influencing factors.

Comparative Responses of Orange-Foot and Common-Foot Haliotis gigantea to Carotenoid-Enriched Diets: Survival, Heat Tolerance, and Bacterial Resistance.

Carotenoids, known to enhance survival, heat tolerance, and bacterial resistance, play an essential role in the nutrition of economically important aquatic animals. This study specifically examined their impact as feed additives on the abalone Haliotis gigantea. We prepared 13 compound feeds with varying levels of astaxanthin, zeaxanthin, and ß-carotene, and administered them to both common-footed and orange-footed H. gigantea. The survival rate of H. gigantea was about 70-80%, with no significant differences in survival observed among the various carotenoid-supplemented feeding groups or when compared with the control group, nor between orange-footed and common-footed individuals. In heat attachment duration experiments, orange-foot abalones exhibited longer attachment durations with certain concentrations of astaxanthin and zeaxanthin, whereas common-foot abalones showed extended durations with astaxanthin, zeaxanthin, and ß-carotene, indicating that common-foot abalones might benefit more from these carotenoids. Additionally, our results showed similar patterns and levels of Vibrio harveyi AP37 resistance in both orange-footed and common-footed H. gigantea, suggesting a uniform response to carotenoid supplementation in their bacterial defense mechanisms. This study suggests the potential benefits of carotenoid supplementation in H. gigantea and contributes to the theoretical basis for developing high-quality artificial compound feeds.

Soft Self-Healing Robot Driven by New Micro Two-Way Shape Memory Alloy Spring.

Soft robotic bodies are susceptible to mechanical fatigue, punctures, electrical breakdown, and aging, which can result in the degradation of performance or unexpected failure. To overcome these challenges, a soft self-healing robot is created using a thermoplastic methyl thioglycolate-modified styrene-butadiene-styrene (MG-SBS) elastomer tube fabricated by melt-extrusion, to allow the robot to self-heal autonomously at room temperature. After repeated damage and being separated into several parts, the robot is able to heal its stiffness and elongation to break to enable almost complete recovery of robot performance after being allowed to heal at room temperature for 24 h. The self-healing capability of the robot is examined across the material scale to robot scale by detailed investigations of the healing process, healing efficiency, mechanical characterization of the robot, and assessment of dynamic performance before and after healing. The self-healing robot is driven by a new micro two-way shape-memory alloy (TWSMA) spring actuator which achieved a crawling speed of 21.6 cm/min, equivalent to 1.57 body length per minute. An analytical model of the robot is created to understand the robot dynamics and to act as an efficient tool for self-healing robot design and optimization. This work therefore provides a new methodology to create efficient, robust, and damage-tolerant soft robots.

Gross ecosystem product (GEP): Quantifying nature for environmental and economic policy innovation.

The large-scale loss of ecosystem assets around the world, and the resultant reduction in the provision of nature's benefits to people, underscores the urgent need for better metrics of ecological performance as well as their integration into decision-making. Gross ecosystem product (GEP) is a measure of the aggregate monetary value of final ecosystem-related goods and services in a specific area and for a given accounting period. GEP accounting captures the use of many ecosystem services in production processes across the economy, which are then valued in terms of their benefits to society. GEP has five key elements that make it transparent, trackable, and readily understandable: (1) a focus on nature's contributions to people; (2) the measurement of ecosystem assets as stocks and ecosystem services as flows; (3) the quantification of ecosystem service use; (4) an understanding of ecosystem service supply chains through value realization; and (5) the disaggregation of benefits across groups. Correspondingly, a series of innovative policies based on GEP have been designed and implemented in China. The theoretical and practical lessons provided by these experiences can support continued policy innovation for green and inclusive development around the world.

Composite dietary antioxidant index and the risk of heart failure: A cross-sectional study from NHANES.

BACKGROUND: Previous studies show that oxidative stress is important in heart failure (HF) pathogenesis. The composite dietary antioxidant index (CDAI), which reflects the antioxidant profile of nutrient supplements, is associated with cardiovascular mortality risk. However, the association between CDAI and the risk of HF remains unknown. HYPOTHESIS: In this study, we investigated the relationship between CDAI and HF risk using National Health and Nutritional Examination Survey (NHANES) data. METHODS: The data of participants aged >40 years old from the NHNANES between 2001 and 2018 were obtained and used to assess the relationship between CDAI and the risk of HF. Logistic regression was used to calculate the odds ratio (OR) of CDAI for the risk of HF. RESULTS: A total of 29 101 participants were divided into the HF (n = 1419; 4.88%) and non-HF groups (n = 27 682; 95.12%), HF group participants had lower CDAI than the non-HF group (-0.32 ± 0.14 vs. 0.67 ± 0.05, p < .0001). Compared with the lowest CDAI quartile (Q1), the OR for HF risk was 0.88 (0.68-1.13) for Q2 (p = .30), 0.77 (0.61-0.99) for Q3 (p = .04), and 0.68 (0.52-0.89) for Q4 (p = .01). CONCLUSIONS: CDAI was negatively associated with the risk of HF. Our findings show that the intake of an antioxidant-rich dietary is a potential method to reduce the risk of HF.

Ecological benefits of artificial light at night (ALAN): Accelerating the development and metamorphosis of marine shellfish larvae.

Urbanization has led to increasing use of artificial light at night (ALAN), which has rapidly become an important source of pollution in many cities. To identify the ALAN effects on the embryonic development of the Pacific abalone Haliotis discus hannai, we first exposed larvae to natural light with a light period of 12 L:12D (control, Group CTR). We then exposed larvae to three different light regimes. Larvae in Group NL were exposed to full spectrum artificial light from 18:00 to 00:00 to simulate the lighting condition at night, whereas Groups BL and YL were illuminated at the same time interval with 450 nm of short-wavelength blue light and 560 nm of long-wavelength orange light, respectively, to simulate billboard lighting at night. There were significantly higher hatching success and metamorphosis rates of larvae in Group BL than in Group YL or CTR (P < 0.05). The larvae in Group YL had the highest abnormality rate and took the longest time to complete metamorphosis. Transcriptomic studies revealed significantly higher expression levels of genes related to RNA transport, DNA replication, and protein processing in endoplasmic reticulum pathways in Group BL compared to the other groups. In the metabolomic analysis, we identified prostaglandin B1, tyramine, d-fructose 6-phosphate, L-adrenaline, leukotriene C4, and arachidonic acid as differential metabolic markers, as they play a vital part in helping larvae adapt to different ALAN conditions. Multi-omics correlation analysis of pairwise comparisons between all of the groups suggested that the biosynthesis of unsaturated fatty acids (FAs) and arachidonic acid metabolism pathways were significantly enriched (P < 0.05). Further quantitative analysis of the fatty acid (FA) contents revealed that 42 out of 50 FAs were down-regulated in Group BL and up-regulated in Group YL, which suggested that the synthesis, catabolism, and metabolism of FAs are crucial for the larval response to different spectral components of ALAN. For the first time, we report positive rather than negative effects of artificial blue light at night on the embryonic development of a benthic marine species. These results are significant for unbiased and full-scale assessment of the ecological effects of ALAN and for understanding the structural stability of the marine benthic community.

Genomic Characterization and gE/gI-Deleted Strain Construction of Novel PRV Variants Isolated in Central China.

Pseudorabies virus (PRV) variants have caused substantial economic losses in the swine industry in China since 2011. To surveil the genetic variation in PRV field strains, here, two novel variant strains of PRV were isolated from Shanxi Province in central China and were designated SX1910 and SX1911. To identify the genetic characteristics of the two isolates, their complete genomes were sequenced, and phylogenetic analysis and sequence alignment revealed that field PRV variants have undergone genetic variations; notably, the protein-coding sequences UL5, UL36, US1 and IE180 exhibited extensive variation and contained one or more hypervariable regions. Furthermore, we also found that the glycoproteins gB and gD of the two isolates had some novel amino acid (aa) mutations. Importantly, most of these mutations were located on the surface of the protein molecule, according to protein structure model analysis. We constructed a mutant virus of SX1911 with deletion of the gE and gI genes via CRISPR/Cas9. When tested in mice, SX1911-ΔgE/gI-vaccinated mice were protected within a comparable range to Bartha-K61-vaccinated mice. Additionally, a higher dose of inactivated Bartha-K61 protected the mice from lethal SX1911 challenge, while a lower neutralization titer, higher viral load and more severe microscopic lesions were displayed in Bartha-K61-vaccinated mice. These findings highlight the need for continuous monitoring of PRV and novel vaccine development or vaccination program design for PRV control in China.

A Smart Responsive Fluorescence-MR Nanoprobe for Monitoring Tumor Response to Immunotherapy.

Accurately evaluating tumor responses to immunotherapy is clinically relevant. However, non-invasive, real-time visualization techniques to evaluate tumor immunotherapy are still lacking. Herein, a smart responsive fluorescence-MR dual-modal nanoprobe, QM(GP)-MZF(CP), is reported that can be targeted for cleavage by the cytotoxic T cell activation marker granzyme B and the apoptosis-related marker cysteine-aspartic acid-specific protease 3 (Caspase-3). The probe uses quinoline-malononitrile (QM), an aggregation-induced emission luminogen, and Mn-Zn ferrite magnetic nanoparticles (MZF-MNPs), a T2-weighted imaging (T2WI) contrast agent, as imaging molecules that are linked with the substrate peptides specific to granzyme B and Caspase-3. Therefore, both granzyme B and Caspase-3 can target and cleave the substrate peptides in QM(GP)-MZF(CP). Via aggregation-induced fluorescence imaging of QM and the aggregation-induced T2WI-enhanced imaging effect of MZF-MNPs, the status of T cells after tumor immunotherapy and the subsequent triggering of tumor cell apoptosis can be determined to identify tumor responsiveness to immunotherapy and thereby evaluate the effectiveness of this therapy in the early stages of treatment.

Magnetic resonance/fluorescence dual-modality contrast agents targeting αvß6-overexpressing tumors based on A20FMDV2 peptide as a ligand.

Pancreatic ductal adenocarcinoma (PDAC) is a malignant digestive system tumor with a poor late-stage prognosis. This study aimed to identify new methods for the early detection of PDAC. The nanoprobe A20FMDV2-Gd-5-FAM was developed using A20FMDV2 (N1AVPNLRGDLQVLAQKVART20-NH2, A20FMDV2) as the ligand and characterized using dynamic light scattering, transmission electron microscopy, Fourier transform infrared analysis, and UV absorption spectroscopy. The binding of pancreatic cancer cells AsPC-1, MIA PaCa-2, and normal human pancreatic H6C7 cells (HPDE6-C7) to the probe was verified using laser confocal microscopy, and the biocompatibility of the probe was evaluated in vivo. In vivo magnetic resonance and fluorescence imaging were also performed on nude mice with subcutaneous pancreatic tumor xenografts to verify the bimodal imaging performance of the probe. The probe exhibited good stability and biocompatibility and an enhanced relaxation rate (25.46 ± 1.32 mM-1 s-1) than Gd-DTPA. Confocal laser scanning microscopy results revealed that the A20FMDV2-Gd-5-FAM probe could be successfully ingested and internalized, and infrared analysis results demonstrated that the probe was linked successfully. Finally, magnetic resonance T1WI imaging and intravital fluorescence imaging demonstrated the specific signal enhancement of the probe at the tumor site. In conclusion, the bimodal molecular probe A20FMDV2-Gd-5-FAM showed a stable magnetic resonance and fluorescence bimodal imaging performance and is a promising new approach for diagnosing early-stage cancers with a high integrin αvß6 expression.

pH/thermal dual-responsive multifunctional drug delivery system for effective photoacoustic imaging-guided tumor chemo/photothermal therapy.

The development of a combination of chemo/photothermal therapy could overcome the limitations of single-modality therapy and enhance therapeutic efficacy. In this study, a pH/thermal dual-responsive multifunctional drug delivery system with dual-drug loading and enhanced chemo/photothermal therapy is developed based on polydopamine-coated mesoporous silica-gold nanorods (PDA-AuNRs@MSN). Nanoscale mesoporous silica-gold nanorods encapsulating doxorubicin (DOX) are designed as a core and then modified by polydopamine. The PDA shell not only conjugates with another anticancer bortezomib (Btz) to form pH-sensitive bond through boronic acid and catechol but also acts as a gatekeeper to control the release of doxorubicin and enhance the photothermal effect. Such a nanocarrier not only acts as a contrast agent for PA imaging but also serves as a therapeutic agent for enhanced chemo/photothermal therapy. The DOX and Btz could be released in an on-demand mode under near-infrared light irradiation and acid environment. The tumor size and location could be observed via PA imaging after intravenous injection into 4T1-bearing mice. Compared with AuNRs@MSN, PDA-AuNRs@MSN exhibit an increased near-infrared (NIR) absorption at 808 nm and an enhanced photothermal effect. The integrated D/B-PDA-AuNRs@MSN nanoparticles show higher cell apoptosis and enhanced tumor treatment efficacy in vitro and in vivo in comparison with single chemotherapy or photothermal therapy. Combined together, D/B-PDA-AuNRs@MSN show pH/thermal-responsive controlled-release and synergistic chemo/photothermal therapy for tumor.

Intracardiac Echocardiography-Guided Implantation for Proximal Left Bundle Branch Pacing.

BACKGROUND: Multiple screw-in attempts under fluoroscopy are often needed to place the pacing lead tip near or at the left bundle branch (LBB). This study was conducted to evaluate the feasibility of implanting an LBB pacing lead in the proximal LBB (PLBB) guided by intracardiac echocardiography (ICE). METHODS: The distribution of the LBB was initially determined by ICE anatomic imaging and 3-dimensional electrical mapping of His and LBB potentials in 20 patients in the first parts of the study. In the second part, 101 consecutive pacemaker-indicated patients were randomized into the ICE-guided and non-ICE groups for LBB pacing implantation. The procedural details and electrophysiological characteristics of the 2 groups were compared. RESULTS: In the first part of the study, PLBB was identified at 10 to 20 mm from the tricuspid annulus toward the apex with an area of 4.5±1.1 cm2. In the second part, the number of lead screw-in attempts in the septum was fewer in the ICE group than in the non-ICE group (1.43±0.62 versus 1.98±0.75, P=0.0002). The duration of the procedure (26±8 versus 43±9 minutes, P<0.001) and fluoroscopy for LBB pacing implantation (7.4±1.8 versus 10.7±2.4 minutes, P<0.001) in the ICE group was significantly shorter than those in the non-ICE group. LBB pacing in the ICE group generated a lesser QRS duration with more cases of LBB trunk pacing (46.8% versus 25%, P=0.031) and PLBB (91.5% versus 72.7%, P=0.0267) pacing compared with that in the non-ICE group. CONCLUSIONS: The basal left ventricular septum can be better visualized using ICE. ICE-guided PLBB pacing is feasible and safe, with a shorter duration required for the procedure and fluoroscopy, and generates greater LBB trunk pacing and PLBB pacing.

Effects of bioaugmentation by isolated Achromobacter xylosoxidans BP1 on PAHs degradation and microbial community in contaminated soil.

Polycyclic aromatic hydrocarbons (PAHs) are a group of organic pollutants ubiquitous and persistent in soil. In order to provide a viable solution for bioremediation of PAHs-contaminated soil, a strain of Achromobacter xylosoxidans BP1 with superior PAHs degradation ability was isolated from contaminated soil at a coal chemical site in northern China. The degradation of phenanthrene (PHE) and benzo[a]pyrene (BaP) by strain BP1 was investigated in three different liquid phase cultures, and the removal rates of PHE and BaP by strain BP1 were 98.47% and 29.86% after 7 days under the conditions of PHE and BaP as the only carbon source, respectively. In the medium with the coexistence of PHE and BaP, the removal rates of BP1 were 89.44% and 9.42% after 7 days, respectively. Then, strain BP1 was investigated for its feasibility in remediating PAH-contaminated soil. Among the four PAHs-contaminated soils treated differently, the treatment inoculated with BP1 exhibited higher removal rates of PHE and BaP (p < 0.05), especially the CS-BP1 treatment (inoculation of BP1 into unsterilized PAHs-contaminated soil) showed 67.72%, 13.48% removal of PHE and BaP, respectively, over 49 days of incubation. Bioaugmentation also significantly increased the activity of dehydrogenase and catalase in the soil (p＜0.05). Furthermore, the effect of bioaugmentation on the removal of PAHs was investigated by measuring the activity of dehydrogenase (DH) and catalase (CAT) during incubation. Among them, the DH and CAT activities of CS-BP1 and SCS-BP1 (inoculation of BP1 into sterilized PAHs-contaminated soil) treatments inoculated with strain BP1 were significantly higher than those of treatments without BP1 addition during incubation (p < 0.01). The structure of the microbial community varied among treatments, but the Proteobacteria phylum showed the highest relative abundance in all treatments of the bioremediation process, and most of the bacteria with higher relative abundance at the genus level also belonged to the Proteobacteria phylum. Prediction of microbial functions in soil by FAPROTAX analysis showed that bioaugmentation enhanced microbial functions associated with the degradation of PAHs. These results demonstrate the effectiveness of Achromobacter xylosoxidans BP1 as a PAH-contaminated soil degrader for the risk control of PAHs contamination.

ACE2 PET to reveal the dynamic patterns of ACE2 recovery in an infection model with pseudocorona virus.

Due to the COVID-19 pandemic, a series of sequelae, such as fatigue, tachypnea, and ageusia, appeared in long COVID patients, but the pathological basis was still uncertain. The targeted radiopharmaceuticals were of potential to systemically and dynamically trace the pathological changes. For the key ACE2 protein in the virus-host interaction, 68 Ga-cyc-DX600 was developed on the basis of DX600 as a PET tracer of ACE2 fluctuation and maintained the ability in differentiating ACE and ACE2. In the temporary infection model inhaled with the radio-traceable pseudovirus in the upper respiratory tract of male humanized ACE2 (hACE2) mice, organ-specific ACE2 dysfunction in acute period and the following ACE2 recovery in a relatively long period was visualized and quantified by ACE2 PET, revealing a complex pattern of virus concentration-dependent degree and time period-dependent tendency of ACE2 recovery, mainly a sudden decrease of apparent ACE2 in the heart, liver, kidneys, lungs, and so on, but the liver was of a quick functional compensation on ACE2 expression after a temporary decrease. ACE2 expression of most organs has recovered to a normal level at 15 days post inhalation, with brain and genitals still of a decreased SUVACE2 ;  meanwhile, kidneys were of an increased SUVACE2 . These findings on ACE2 PET were further verified by western blot. When compared with high-resolution computed tomography on structural changes and FDG PET on glycometabolism, ACE2 PET was superior in an earlier diagnostic window during infection and more comprehensive understanding of functional dysfunction post-infection. In the respective ACE2 PET/CT and ACE2 PET/MR scans of a volunteer, the repeatability of SUVACE2 and the ACE2 specificity were further confirmed. In conclusion, 68 Ga-cyc-DX600 was developed as an ACE2-specific tracer, and the corresponding ACE2 PET revealed the dynamic patterns of functional ACE2 recovery and provided a reference and approach to explore the ACE2-related pathological basis of sequelae in long COVID.

Structural characteristics of orexin receptor type 2 in Pacific abalone and its diurnal expression pattern after fasting and re-feeding.

Pacific abalone (Haliotis discus hannai) is a typical nocturnal organism. To examine the circadian expression pattern of orexin receptor type 2 (OX2R) and its potential effect on the feeding behavior of abalone, the coding region sequence of OX2R that is 1215 bp in length and encodes 404 amino acids was first cloned using the rapid amplification of cDNA ends technique. A recombinant expression vector was constructed for H. discus hannai based on the OX2R protein, obtaining a recombinant protein with a molecular weight of 46 kDa. Polyclonal antibody was prepared with the purified recombinant protein used as the antigen, and the antibody titer of ≥512 K was detected by enzyme-linked immunosorbent assay. The expression levels of OX2R determined using western blotting were highest in the intestinal tract (P < 0.05), but they were not significantly different from the levels in the pedal. Immunofluorescence experiments affirmed that OX2R was widely expressed in the columnar cells of the intestinal mucosal epithelium. To further account for the relationship between the onset of feeding behavior and the expression level of OX2R in abalone, the circadian expression characteristics of OX2R were analyzed by dissecting the intestinal tissues after three days of normal feeding and fasting and following the refeeding treatment. The expression levels of OX2R in the refeeding group were significantly higher than those in the normal feeding and fasting groups at any time point (P < 0.05). The cosine curve analysis revealed that the expression levels of OX2R lost rhythmicity after fasting. Based on the quantification of behavioral data for abalone after fasting and refeeding, the cumulative movement distance and movement duration in each group followed a significant cosine rhythm (P < 0.05), which is consistent with abalone's nocturnal ecological habits. However, the cumulative movement distance and movement duration in the fasting group were significantly lower than those in the normal feeding and refeeding groups (P < 0.05). The peak phases of the cumulative movement distance and movement duration in the refeeding group (ZT08:22 and ZT08:44) shifted backward compared to the normal feeding group (ZT07:33 and ZT07:39). The above results first identified the structural characteristics and circadian expression patterns of OX2R in the marine mollusk abalone, which may reveal the molecular mechanism behind the generation of a feeding rhythm in marine nocturnal organisms and serve as a tool helping to maintain the diversity of marine benthic resources.

Reproduction strategy of nocturnal marine molluscs: running for love.

The cost of reproduction is the core driver of life history evolution in animals. This paper demonstrates that the cumulative distance moved and the duration of movement of sexually immature abalones, Haliotis discus hannai, kept in various male and female groups, were significantly higher than those of sexually mature individuals, except when kept in mixed cultures of mature males and females. After mixed-culture, sexually mature males moved significantly further and for a longer duration than mature female abalones, and even more so than mature male abalones of any other group. Examination of the LC-MS metabolomics of mature males cultured with sexually mature females (AM) and those cultured with sexually immature females (JM) showed that cyclic adenosine monophosphate (cAMP) acted as a differential metabolic biomarker. After 24-h uninterrupted sampling, the concentration of 5-HT and the expression levels of the 5-HT2 and 5-HT6 receptors in AM were significantly higher than those in JM. After further injection of 5-HT2 and 5-HT6 receptor antagonists, the concentrations of cAMP and PKA rose again, but the cumulative movement duration and distance of male abalones decreased significantly, showing that 5-HT was involved in the regulation of movement behavior of male abalones through the 5-HT2 and 5-HT6 receptor-activated cAMP-PKA pathways. The results demonstrated a significant increase in the movement endurance of mature male abalones cultured with mature females, providing a theoretical basis for understanding the adaptive life history strategies of abalones and suggesting ways to protect diverse benthic resources for abalones during the reproductive stage.