A bivalent ß-carboline derivative inhibits macropinocytosis-dependent entry of pseudorabies virus by targeting the kinase DYRK1A.

Pseudorabies virus (PRV) has become a "new life-threatening zoonosis" since the human-originated PRV strain was first isolated in 2020. To identify novel anti-PRV agents, we screened a total of 107 ß-carboline derivatives and found 20 compounds displaying antiviral activity against PRV. Among them, 14 compounds showed better antiviral activity than acyclovir. We found that compound 45 exhibited the strongest anti-PRV activity with an IC50 value of less than 40 nM. Our in vivo studies showed that treatment with 45 significantly reduced the viral loads and protected mice challenged with PRV. To clarify the mode of action of 45, we conducted a time of addition assay, an adsorption assay, and an entry assay. Our results indicated that 45 neither had a virucidal effect nor affected viral adsorption while significantly inhibiting PRV entry. Using the FITC-dextran uptake assay, we determined that 45 inhibits macropinocytosis. The actin-dependent plasma membrane protrusion, which is important for macropinocytosis, was also suppressed by 45. Furthermore, the kinase DYRK1A (dual-specificity tyrosine phosphorylation-regulated kinase 1A) was predicted to be a potential target for 45. The binding of 45 to DYRK1A was confirmed by drug affinity responsive target stability and cellular thermal shift assay. Further analysis revealed that knockdown of DYRK1A by siRNA suppressed PRV macropinocytosis and the tumor necrosis factor alpha-TNF-induced formation of protrusions. These results suggested that 45 could restrain PRV macropinocytosis by targeting DYRK1A. Together, these findings reveal a unique mechanism through which ß-carboline derivatives restrain PRV infection, pointing to their potential value in the development of anti-PRV agents.

9-Butyl-Harmol Exerts Antiviral Activity against Newcastle Disease Virus through Targeting GSK-3ß and HSP90ß.

The paramyxoviruses represent a large family of human and animal pathogens that cause significant health and economic burdens worldwide. However, there are no available drugs against the virus. ß-carboline alkaloids are a family of naturally occurring and synthetic products with outstanding antiviral activities. Here, we examined the antiviral effect of a series of ß-carboline derivatives against several paramyxoviruses, including Newcastle disease virus (NDV), peste des petits ruminants virus (PPRV), and canine distemper virus (CDV). Among these derivatives, 9-butyl-harmol was identified as an effective antiviral agent against these paramyxoviruses. Further, a genome-wide transcriptome analysis in combination with target validation strategies reveals a unique antiviral mechanism of 9-butyl-harmol through the targeting of GSK-3ß and HSP90ß. On one hand, NDV infection blocks the Wnt/ß-catenin pathway to suppress the host immune response. 9-butyl-harmol targeting GSK-3ß dramatically activates the Wnt/ß-catenin pathway, which results in the boosting of a robust immune response. On the other hand, NDV proliferation depends on the activity of HSP90. The L protein, but not the NP protein or the P protein, is proven to be a client protein of HSP90ß, rather than HSP90α. 9-butyl-harmol targeting HSP90ß decreases the stability of the NDV L protein. Our findings identify 9-butyl-harmol as a potential antiviral agent, provide mechanistic insights into the antiviral mechanism of 9-butyl-harmol, and illustrate the role of ß-catenin and HSP90 during NDV infection. IMPORTANCE Paramyxoviruses cause devastating impacts on health and the economy worldwide. However, there are no suitable drugs with which to counteract the viruses. We determined that 9-butyl-harmol could serve as a potential antiviral agent against paramyxoviruses. Until now, the antiviral mechanism of ß-carboline derivatives against RNA viruses has rarely been studied. Here, we found that 9-butyl-harmol exerts dual mechanisms of antiviral action, with its antiviral activities being mediated by two targets: GSK-3ß and HSP90ß. Correspondingly, the interaction between NDV infection and the Wnt/ß-catenin pathway or HSP90 is demonstrated in this study. Taken together, our findings shed light on the development of antiviral agents against paramyxoviruses, based on the ß-carboline scaffold. These results present mechanistic insights into the polypharmacology of 9-butyl-harmol. Understanding this mechanism also deepens the host-virus interaction and reveals new drug targets for anti-paramyxoviruses.

Major quality regulation network of flavonoid synthesis governing the bioactivity of black wolfberry.

Black wolfberry (Lycium ruthenicum Murr.) contains various bioactive metabolites represented by flavonoids, which are quite different among production regions. However, the underlying regulation mechanism of flavonoid biosynthesis governing the bioactivity of black wolfberry remains unclear. Presently, we compared the bioactivity of black wolfberry from five production regions. Multi-omics were performed to construct the regulation network associated with the fruit bioactivity. The detailed regulation mechanisms were identified using genetic and molecular methods. Typically, Qinghai (QH) fruit exhibited higher antioxidant and anti-inflammatory activities. The higher medicinal activity of QH fruit was closely associated with the accumulation of eight flavonoids, especially Kaempferol-3-O-rutinoside (K3R) and Quercetin-3-O-rutinoside (rutin). Flavonoid biosynthesis was found to be more active in QH fruit, and the upregulation of LrFLS, LrCHS, LrF3H and LrCYP75B1 caused the accumulation of K3R and rutin, leading to high medicinal bioactivities of black wolfberry. Importantly, transcription factor LrMYB94 was found to regulate LrFLS, LrCHS and LrF3H, while LrWRKY32 directly triggered LrCYP75B1 expression. Moreover, LrMYB94 interacted with LrWRKY32 to promote LrWRKY32-regulated LrCYP75B1 expression and rutin synthesis in black wolfberry. Transgenic black wolfberry overexpressing LrMYB94/LrWRKY32 contained higher levels of K3R and rutin, and exhibited high medicinal bioactivities. Importantly, the LrMYB94/LrWRKY32-regulated flavonoid biosynthesis was light-responsive, showing the importance of light intensity for the medicinal quality of black wolfberry. Overall, our results elucidated the regulation mechanisms of K3R and rutin synthesis, providing the basis for the genetic breeding of high-quality black wolfberry.

3-Amide-ß-carbolines block the cell cycle by targeting CDK2 and DNA in tumor cells potentially as anti-mitotic agents.

ß-Carboline alkaloids are natural and synthetic products with outstanding antitumor activity. C3 substituted and dimerized ß-carbolines exert excellent antitumor activity. In the present research, 37 ß-carboline derivatives were synthesized and characterized. Their cytotoxicity, cell cycle, apoptosis, and CDK2- and DNA-binding affinity were evaluated. ß-Carboline monomer M3 and dimer D4 showed selective activity and higher cytotoxicity in tumor cells than in normal cells. Structure-activity relationships (SAR) indicated that the amide group at C3 enhanced the antitumor activity. M3 blocked the A549 (IC50 = 1.44 ± 1.10 µM) cell cycle in the S phase and inhibited A549 cell migration, while D4 blocked the HepG2 (IC50 = 2.84 ± 0.73 µM) cell cycle in the G0/G1 phase, both of which ultimately induced apoptosis. Furthermore, associations of M3 and D4 with CDK2 and DNA were proven by network pharmacology analysis, molecular docking, and western blotting. The expression level of CDK2 was downregulated in M3-treated A549 cells and D4-treated HepG2 cells. Moreover, M3 and D4 interact with DNA and CDK2 at sub-micromolar concentrations in endothermic interactions caused by entropy-driven adsorption processes, which means that the favorable entropy change (ΔS > 0) overcomes the unfavorable enthalpy change (ΔH > 0) and drives the spontaneous reaction (ΔG < 0). Overall, these results clarified the antitumor mechanisms of M3 and D4 through disrupting the cell cycle by binding DNA and CDK2, which demonstrated the potential of M3 and D4 as novel antiproliferative drugs targeting mitosis.

Large-Area Black Phosphorus/PtSe2 Schottky Junction for High Operating Temperature Broadband Photodetectors.

High operating temperature (HOT) broadband photodetectors are urgently necessary for extreme condition applications in infrared-guided missiles, infrared night vision, fire safety imaging, and space exploration sensing. However, conventional photodetectors show dramatic carrier mobility decreases and carrier losses with low photoresponsivity at HOT due to the increased carrier scattering in channels at high temperatures. Herein, the HOT broadband photodetectors from room temperature to 470 K are developed for the first time by large-area black phosphorus (BP)/PtSe2 films device arrays via a depletion-enhanced photocarrier dynamics strategy. Attributed to the 2D Schottky junction at BP/PtSe2 interface and resulting in full depleted working channels, the BP/PtSe2 photodetector arrays exhibit high tolerance to carrier mobility decrease during the increasing operating temperature in a wide wavelength range from 532 to 2200 nm. Thus, the photodetector shows a state-of-the-art operating temperature at 470 K with the photo-responsivity (R) and specific detectivity (D*) of 25 A W-1 and 6.4 × 1011 Jones under 1850 nm illumination, respectively. Moreover, BP/PtSe2 photodetector arrays show high-uniformity photo-response in a large area. This work provides new strategies for high-performance broadband photodetector arrays with HOT by Schottky junction of large-area BP/PtSe2 films.

Major histocompatibility complex class IIα and IIß of pufferfish (Takifugu obscurus): Identification and functional characterization in response to bacterial challenge.

Classical major histocompatibility complex (MHC) class II molecules play an essential role in immune system. In this study, MHC IIα (Pf-MHC IIα) and MHC IIß (Pf-MHC IIß) homology genes from pufferfish (Takifugu obscurus) were cloned and their functional characterization in response to bacterial challenge was identified. The nucleotide sequences of the open reading frames (ORFs) of pufferfish Pf-MHC IIα and Pf-MHC IIß were 708 bp and 750 bp, encoding 235 aa and 249 aa, respectively. The structure of Pf-MHC IIα or Pf-MHC IIß contained a signal peptide, an α1/ß1 domain, an α2/ß2 domain, a transmembrane region and a cytoplasmic region. Multiple sequence alignment and phylogenetic analysis showed that Pf-MHC IIα and Pf-MHC IIß molecules had the highest similarity with Fugu rubripes (Takifugu rubripes). Cellular localization analysis indicated that the distribution of Pf-MHC IIα and Pf-MHC IIß was in the lymphocyte membrane and cytoplasm. qRT-PCR results showed that Pf-MHC IIα and Pf-MHC IIß expressed relatively high in skin, gills and gut. In addition, after stimulation challenge in vitro (lipopolysaccharide, or polyinosinic: polycytidylic acid) and in vivo (A. hydrophila), the mRNA expressions of Pf-MHC IIα and Pf-MHC IIß were significantly up-regulated in lymphocytes and in tissues of skin, gills, gut and head kidney. Moreover, Pf-MHC IIα or Pf-MHC IIß neutralization reduced the ability of A. hydrophila to induce the expressions of lymphocyte cytokines (TNF-α, IL-1ß and IL-10). Overall, it is speculated that Pf-MHC IIα and Pf-MHC IIß may play an important role in the host response against A. hydrophila in pufferfish.

The Detection of Yarn Roll's Margin in Complex Background.

Online detection of yarn roll's margin is one of the key issues in textile automation, which is related to the speed and scheduling of bobbin (empty yarn roll) replacement. The actual industrial site is characterized by uneven lighting, restricted shooting angles, diverse yarn colors and cylinder yarn types, and complex backgrounds. Due to the above characteristics, the neural network detection error is large, and the contour detection extraction edge accuracy is low. In this paper, an improved neural network algorithm is proposed, and the improved Yolo algorithm and the contour detection algorithm are integrated. First, the image is entered in the Yolo model to detect each yarn roll and its dimensions; second, the contour and dimensions of each yarn roll are accurately detected based on Yolo; third, the diameter of the yarn rolls detected by Yolo and the contour detection algorithm are fused, and then the length of the yarn rolls and the edges of the yarn rolls are calculated as measurements; finally, in order to completely eliminate the error detection, the yarn consumption speed is used to estimate the residual yarn volume and the measured and estimated values are fused using a Kalman filter. This method overcomes the effects of complex backgrounds and illumination while being applicable to different types of yarn rolls. It is experimentally verified that the average measurement error of the cylinder yarn diameter is less than 8.6 mm, and the measurement error of the cylinder yarn length does not exceed 3 cm.

Influence of modified biochar supported sulfidation of nano-zero-valent-iron (S-nZVI/BC) on nitrate removal and greenhouse gas emission in constructed wetland.

In this study, the biochar (BC) produced from sawdust, sludge, reed and walnut were used to support sulfidation of nano-zero-valent-iron (S-nZVI) to enhance nitrate (NO3--N) removal and investigate the impact on greenhouse gas emissions. Batch experiment results showed the S-nZVI/BCsawdust (2:1, 500), S-nZVI/BCsludge (2:1, 900), S-nZVI/BCreed (2:1, 700), and S-nZVI/BC walnut (2:1, 700) respectively improved NO3--N removal efficiencies by 22%, 20%, 3% and 0.1%, and the selectivity toward N2 by 22%, 25%, 22% and 18%. S-nZVI uniformly loaded on BC provided electrons for the conversion of NO3--N to N2 through Fe0. At the same time, FeSx layer was formed on the outer layer of ZVI in the sulfidation process to prevent iron oxidation, so as to improve the electrons utilization efficiency After adding four kinds of S-nZVI/BC into constructed wetlands (CWs), the NO3--N removal efficiencies could reach 100% and the N2O emission fluxes were reduced by 24.17%-36.63%. And the average removal efficiencies of TN, COD, TP were increased by 21.9%, -16.5%, 44.3%, repectively. The increasing relative abundances of denitrifying bacteria, such as Comamonas and Simplicispira, suggested that S-nZVI/BC could also improve the process of microbial denitrification. In addition, different S-nZVI/BC had different effects on denitrification functional genes (narG, nirk, nirS and nosZ genes), methanotrophs (pmoA) and methanogenesis (mcrA). This research provided an effective method to improve NO3--N removal and reduce N2O emission in CWs.

Dynamic mRNA and miRNA expression of the head during early development in bighead carp (Hypophthalmichthys nobilis).

BACKGROUND: Head of fish species, an exquisitely complex anatomical system, is important not only for studying fish evolution and development, but also for economic values. Currently, although some studies have been made on fish growth and body shapes, very limited information is available on the molecular mechanism of head development. RESULTS: In this study, RNA sequencing (RNA-Seq) and small RNA sequencing (sRNA-Seq) technologies were used to conduct integrated analysis for the head of bighead carp at different development stages, including 1, 3, 5, 15 and 30 Dph (days post hatch). By RNA-Seq data, 26 pathways related to growth and bone formation were identified as the main physiological processes during early development. Coupling this to sRNA-Seq data, we picked out six key pathways that may be responsible for head development, namely ECM receptor interaction, TNF signaling pathway, osteoclast differentiation, PI3K-Akt signaling pathway, Neuroactive ligand-receptor interaction and Jak-STAT signaling pathway. Totally, 114 important candidate genes from the six pathways were obtained. Then we found the top 20 key genes according to the degree value by cytohubba, which regulated cell growth, skeletal formation and blood homeostasis, such as pik3ca, pik3r1, egfr, vegfa, igf1 and itga2b. Finally, we also acquired 19 key miRNAs playing multiple roles in the perfection of various tissues in the head (such as brain, eye and mouth) and mineralization of head bone system, such as let-7e, miR-142a-5p, miR-144-3p, miR-23a-3p and miR-223. CONCLUSIONS: Results of this study will be informative for genetic mechanisms of head development and also provide potential candidate targets for the interaction regulation during early growth in bighead carp.

Psychological Status, Compliance, Serum Brain-Derived Neurotrophic Factor, and Nerve Growth Factor Levels of Patients with Depression after Augmented Mindfulness-Based Cognitive Therapy.

Objective: Mindfulness-based cognitive therapy (MBCT) is a cost-effective psychosocial program that prevents relapse/recurrence in major depression. The present study aimed to analyze the effects of augmented MBCT along with standard treatment dominated by pharmacotherapy on psychological state, compliance, brain-derived neurotrophic factor (BDNF), and nerve growth factor (NGF) expression levels in patients with depression. Methods: A total of 160 eligible patients with depression in The First Affiliated Hospital of Zhengzhou University were included in this study. The study randomly assigned the patients to the experimental group (n = 80) and control group (n = 80). All participants were assessed with the questionnaires including the 17-item Hamilton Depression Rating Scale (HAMD-17), Rosenberg Self-esteem Scale (RSES), Self-Acceptance Questionnaire (SAQ), and Stigma Scale (Scale of Stigma in People with Mental Illness, SSPM). The serum levels of BDNF and NGF were detected by enzyme-linked immunosorbent assay (ELISA). Results: After 8 weeks of treatment, the experimental group showed significant lower HAMD-17 score, higher RSES, and SAQ score, as well as lower SSPM score compared with the control group (P < 0.01). Furthermore, ELISA revealed that the serum levels of BDNF and NGF remarkably increased in the experimental group after treatment (P < 0.001). Conclusions: Our data showed that augmented MBCT combined with pharmacotherapy contributed to improvement on patients' psychological state, compliance, and disease recurrence.

Enhancement of wastewater treatment under low temperature using novel electrochemical active biofilms constructed wetland.

A novel electrochemical active biofilms constructed wetland (NEAB-CW) was built to enhance the treatment efficiency for domestic sewage under low temperature environment (0-15 °C). In NEAB-CW, the traditional matrixes were replaced with conductive layer, in which laid stainless steel mesh tubes (SSMT) and added slow-release oxygen matrixes (SROM) and zero-valent iron rod (IR) were used to build a bioelectrochemical activity biofilms system. According to the results of 180 d experiment, the removal efficiencies of COD, NH4+-N and TP of NEAB-CW were 1.52 and 2.21, 2.97 and 1.68, 3.95 and 1.76 times higher than the CW without SROM and IR at 10-20 and 0-10 °C, respectively. The transverse and longitudinal electric potential (EP) variations in NEAB-CW improved microbial activities under low temperature by enhancing the electron transfer efficiency, resulting in higher and stable EP and electron currents density, as well as protein-like contents secreted from biofilms. The pollutant-degrading microorganisms (e.g., Clostridia, Simplicispira), low temperature-resistant microorganisms (e.g., Psychrobacter, Acinetobacter), and electrochemical active microorganisms (e.g., Negativicutes, Gammaproteobacteria) obviously accumulated in NEAB-CW under low temperature environment to generate electricity and degrade pollutants. The results provided a good choice to treat domestic sewage at 0-15 °C by using NEAB-CW.

Cellular and non-target metabolomics approaches to understand the antifungal activity of methylaervine against Fusarium solani.

Botanical fungicides are promising replacements for pure chemical synthetic pesticides in agriculture and organic food production. Methylaervine with good physicochemical properties exhibited effective activity against F. solani (EC50 = 10.56 µM) better than the positive control thiophanate-methyl (EC50 = 27.94 µM). The activity changes of malondialdehyde (MDA), catalase (CAT) and superoxide dismutase (SOD) showed that methylaervine could significantly induce lipid peroxidation and activate the antioxidant enzymes. According to the metabolomics analysis, fifty-one differential metabolites and two major antifungal-related pathways covering tricarboxylic acid (TCA) cycle and steroid biosynthesis were identified. Moreover, the disturbance for TCA cycle was validated by the activity changes of dehydrogenase (MDH) and succinate dehydrogenase (SDH) as well as docking simulation. Homology modeling and docking study revealed that hydrogen bonds and hydrophobic interactions played a vital role in methylaervine-protein stability. This study provided new insight into the antifungal activity of methylaervine, which is important for the development of novel botanical fungicides based on methylaervine.

Molecular cloning, characterization and expression analysis of major histocompatibility complex class I alpha gene of pufferfish (Takifugu obscurus).

The major histocompatibility complex (MHC) genes play a key role in immune response in vertebrates. In this study, an MHC I alpha homolog gene (PfMHC Ⅰα) from pufferfish (Takifugu obscurus) was identified and its subcellular localization and expression patterns of PfMHC Ⅰα after challenge in vivo and in vitro were analysed. The open reading frame of PfMHC Ⅰα was 1,089 bp in length, encoding 362 aa. The immunofluorescence result revealed that PfMHC Ⅰα was presented on the membrane of lymphocytes. qRT-PCR analysis indicated that PfMHC Ⅰα was expressed in all examined tissues, with the highest expression in skin, followed by the expression in gills and whole blood. After challenge of Aeromonas hydrophila or polyinosinic: polycytidylic acid (Poly I:C) in vitro, the expression levels of PfMHC Ⅰα on pufferfish kidney lymphocytes were significantly up-regulated, with the highest expression level at 48 hr post-challenge. After infection with A. hydrophila or Poly I:C in vivo, the expression levels of PfMHC Ⅰα in the skin, whole blood and kidneys were significantly up-regulated. Taken together, it is speculated that PfMHC Ⅰα associates with resistance to both intracellular and extracellular antigens and plays an important role in the host response against pathogen infection in pufferfish.

The Detection of Thread Roll's Margin Based on Computer Vision.

The automatic detection of the thread roll's margin is one of the kernel problems in the textile field. As the traditional detection method based on the thread's tension has the disadvantages of high cost and low reliability, this paper proposes a technology that installs a camera on a mobile robot and uses computer vision to detect the thread roll's margin. Before starting, we define a thread roll's margin as follows: The difference between the thread roll's radius and the bobbin's radius. Firstly, we capture images of the thread roll's end surface. Secondly, we obtain the bobbin's image coordinates by calculating the image's convolutions with a Circle Gradient Operator. Thirdly, we fit the thread roll and bobbin's contours into ellipses, and then delete false detections according to the bobbin's image coordinates. Finally, we restore every sub-image of the thread roll by a perspective transformation method, and establish the conversion relationship between the actual size and pixel size. The difference value of the two concentric circles' radii is the thread roll's margin. However, there are false detections and these errors may be more than 19.4 mm when the margin is small. In order to improve the precision and delete false detections, we use deep learning to detect thread roll and bobbin's radii and then can calculate the thread roll's margin. After that, we fuse the two results. However, the deep learning method also has some false detections. As such, in order to eliminate the false detections completely, we estimate the thread roll's margin according to thread consumption speed. Lastly, we use a Kalman Filter to fuse the measured value and estimated value; the average error is less than 5.7 mm.

Interactions of chlorpyrifos degradation and Cd removal in iron-carbon-based constructed wetlands for treating synthetic farmland wastewater.

Pesticide and heavy metal contaminants, such as chlorpyrifos (CP) and cadmium (Cd) in farmland drainage had caused the water pollution and attracted extensive concerns around the world. The incorporation of zeolite-based iron-carbon (ZB-IC) into constructed wetlands (CWs) was prepared to simultaneously remove chlorpyrifos (CP) and cadmium (Cd) in farmland drainage, and the interaction of CP degradation and Cd removal was investigated. Laboratory simulated experiments were carried out in this study, and the results presented that the removal efficiencies of CP and Cd by ZB-IC coupled CWs (ZB-IC-CW) were 99.55% and 98.59%, respectively, which were much higher than that of the zeolite-based (ZB) CWs (CP = 92.99%; Cd = 63.54%). The removal mechanism of CP and Cd by ZB-IC substrate was mainly attributed to electron transfer, which occurred from iron corrosion and hydrogen generation process. In addition, CP could act as carbon source to promote denitrification process. Microbial analysis revealed that the relative abundances of CP-resistant bacteria (Firmicutes, Clostridia and Acetobacterium), Cd-resistant bacteria (Bacteroidetes) and denitrifying bacteria (Proteobacteria and Patescibacteria) were dramatically increased due to the addition of ZB-IC. The higher czcA gene and opd gene in ZB-IC-CW demonstrated that the addition of CP played a positive role in Cd removal, while Cd showed slightly affect to CP removal.

Identification and functional characterization of three caspases in Takifugu obscurus in response to bacterial infection.

Caspases are evolutionarily conserved proteases, which are inextricably linked with the apoptosis and immune system in mammals. However, the expression pattern and function of some caspases remain largely unknown in pufferfish. In this study, three different pufferfish caspases (caspase-2 (Pfcasp-2), caspase-3 (Pfcasp-3), and caspase-8 (Pfcasp-8)) were characterized, and their expression patterns and functions were determined following Aeromonas hydrophila infection. The open reading frames of Pfcasp-2, -3, and -8 are 1,320, 846, and 1455 bp, respectively. Analyses of sequence alignment and phylogenetic tree showed that casp-2, -3, and -8 share 52%-65%, 33%-40%, 63%-78% overall sequence identities with those of other vertebrates, respectively. 3D structures of Pfcasp-2, -3, and -8 enjoy conservation in core area together, while each owns a distinctive profile. Comparisons of deduced amino acid sequences indicated that Pfcaspases possessed the caspase domain and conserved active sites like 'HG' and 'QACXG' (X for R or G). qRT-PCR results revealed that Pfcasp-2, -3, and -8 were expressed constitutively in a wide range of organs, especially in immune-related organs including whole blood and kidney. In vitro, the expressions of the three caspases (Pfcasp-2, 3, and -8) and immune-related genes (IgM and IL-8) were significantly up-regulated in kidney leukocytes after A. Hydrophila challenge and inhibitors treatment. The expressions of Pfcasp-2 and Pfcasp-3 were successfully inhibited in the kidney leukocytes by Ac-DEVD-CHO (an inhibitor to caspase-3), but the expression of Pfcasp-8 was not affected. Cellular localization analysis showed that the distribution of Pfcasp-2, -3, and -8 was in cytoplasm. Further, overexpression of Pfcasp-2, -3, or -8 was found to cause DNA damage and apoptosis, suggesting that three caspases may be related to apoptosis and mediate different apoptosis pathways in pufferfish. Moreover, the expressions of these caspases were also up-regulated in whole blood and kidney after A. hydrophila challenge, indicating their possible involvement in the immune response against A. hydrophia stimulation. Taken together, the results of this study suggest that the caspase-2,-3, and -8 may play an important role in the apoptosis and immune response in pufferfish.

High-efficiency helium separation through an inorganic graphenylene membrane: a theoretical study.

The rising demand for helium resources makes the effective separation of helium from natural gas increasingly important in the cryogenics industry and welding technology. However, most commonly used membranes cannot efficiently separate helium from the small molecules in natural gas. In this work, using first-principles calculations, combined with molecular dynamics simulations, we showed that efficient separation of helium from natural gas molecules (H2O, CO2, CO, CH4, and N2) as well as noble gas molecules (Ne and Ar) can be achieved in an inorganic graphenylene (IGP) membrane with high selectivities. In particular, molecular dynamics simulations demonstrated that high helium permeance (approximately 10-4 mol m-2 s-1 Pa-1) can be achieved over a wide range of temperatures (100 to 500 K) with high selectivity over other gas molecules. The high permeance and selectivity of the IGP monolayer membrane to helium are quite promising for industrial applications.

Recent progress of antibacterial natural products: Future antibiotics candidates.

The discovery of novel antibacterial molecules plays a key role in solving the current antibiotic crisis issue. Natural products have long been an important source of drug discovery. Herein, we reviewed 256 natural products from 11 structural classes in the period of 2016-01/2020, which were selected by SciFinder with new compounds or new structures and MICs lower than 10 µg/mL or 10 µM as criterions. This review will provide some effective antibacterial lead compounds for medicinal chemists, which will promote the antibiotics research based on natural products to the next level.

Clinicopathological features of IgG4-related kidney disease
.

AIMS: To explore the clinicopathological features of IgG4-related kidney disease (IgG4-RKD) in Chinese patients. MATERIALS AND METHODS: We retrospectively analyzed the clinicopathological features of 7 patients with IgG4-RKD and reviewed relevant literature. RESULTS: There were 7 patients (4 males and 3 females) aged 41 - 78 years (median age: 57.1 years), with a mean affliction of 2.6 organs per patient. All 7 patients had elevated serum IgG4 levels, and most of them had acute or chronic renal insufficiency. Two patients had positive myeloperoxidase-antineutrophil cytoplasmic antibodies (MPO-ANCA), and another 2 patients had positive serum antinuclear antibodies (ANA). All 7 patients had tubulointerstitial nephritis (TIN) with IgG4+ plasma cell infiltration, 3 patients also had glomerular lesions, 2 patients had ANCA-associated glomerulonephritis (ANCA-GN), and the remaining patient had membranous nephropathy (MN). All these patients were treated with glucocorticoids, and 3 of them were given cyclophosphamide simultaneously. Six patients achieved remission and had improved renal function, while 1 patient was on maintenance dialysis. CONCLUSION: IgG4-RKD should be diagnosed by combining the clinical data and renal pathological changes, and it should be differentiated from a variety of diseases. Although ANCA-associated vasculitis (AAV) and IgG4-related disease are distinguishable in most cases, coexistence of ANCA-GN and IgG4-RKD should be considered in some special cases.

Aliifodinibius salipaludis sp. nov., Isolated from Saline-Alkaline Soil.

A Gram-staining-negative, oblong or rod-shaped, non-flagellated and facultatively anaerobic bacterium, designated WN023T, was isolated from natural saline-alkali wetland soil of Binhai new district, Tianjin, China. The colonies of strain WN023T on marine agar 2216 (MA) have a brown, circular, wet pigmentation, and its cells are approximately 1-1.5 µm × 2.0-2.5 µm in size. Growth occurred at 15-45 °C, but optimally at 33-37 °C, in the presence of 5.0-25.0% (w/v) NaCl, but optimally at 10.0-15.0% (w/v) NaCl, and at a pH 6.5-11.0, but optimally at 7.5-8.0. Phylogenetic analysis of the 16S rRNA gene indicates that strain WN023T is a member of the genus Aliifodinibius within the family Balneolaceae, and the 16S rRNA gene sequence similarity of strain WN023T with its close relative Aliifodinibius halophilus KCTC 42497T, Aliifodinibius salicampi KACC 19060T, Aliifodinibius roseus KCTC 23442T, Aliifodinibius sediminis ACCC 10714T, and Fodinibius salinus ACCC 10716T was 97.3%, 94.5%, 93.9%, 93.71%, and 92.7%, respectively. Strain WN023T contained menaquinone-7 (MK-7) as the predominant menaquinone, and iso C15:0, Sum in feature 3 (C16:1ω6c and/or C16:1ω7c), and iso C17:1ω9c as the major fatty acids. The major polar lipids were glycolipid (GL), aminolipid (AL), phospholipid (PL), phosphatidylcholine (PC), phosphatidylethanolamine (PE), diphosphatidylglycerol (DPG), and lipid (L). The size of the draft genome as presented here was 3,583,276 bp in size, and the G+C content is 42.2 mol%. These combined physiological, biochemical, phylogenetic, and genotypic data supported placement of strain WN023T in the genus Aliifodinibius and indicated that it was distinct from all other members in the genus Aliifodinibius. This was also confirmed by the low DNA-DNA hybridization values (38.4%) between strain WN023T and the most closely related recognized Aliifodinibius species-A. halophilus KCTC 42497T. Therefore, we propose a novel species in the genus Aliifodinibius to accommodate the novel isolate: Aliifodinibius salipaludis sp. nov. (type strain WN023T = KCTC 52855T = ACCC 19978T).