Mass Spectrometry Imaging Reveals the Morphology-Dependent Toxicological Effects of Nanosilvers on Multiple Organs of Adult Zebrafish (Danio rerio).

Nanosilvers with multifarious morphologies have been extensively used in many fields, but their morphology-dependent toxicity toward nontarget aquatic organisms remains largely unclear. Herein, we used matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) to investigate the toxicological effects of silver nanomaterials with various morphologies on spatially resolved lipid profiles within multiple organs in adult zebrafish, especially for the gill, liver, and intestine. Integrated with histopathology, enzyme activity, accumulated Ag contents and amounts, as well as MSI results, we found that nanosilvers exhibit morphology-dependent nanotoxicity by disrupting lipid levels and producing oxidative stress. Silver nanospheres (AgNSs) had the highest toxicity toward adult zebrafish, whereas silver nanoflakes (AgNFs) exhibited greater toxicity than silver nanowires (AgNWs). Levels of differential phospholipids, such as PC, PE, PI, and PS, were associated with nanosilver morphology. Notably, we found that AgNSs induced greater toxicity in multiple organs, such as the brain, gill, and liver, while AgNWs and AgNFs caused greater toxicity in the intestine than AgNSs. Lipid functional disturbance and oxidative stress further caused inflammation and membrane damage after exposure to nanosilvers, especially with respect to sphere morphology. Taken together, these findings will contribute to clarifying the toxicological effects and mechanisms of different morphologies of nanosilvers in adult zebrafish.

Prediction of compressive comfort of graded compression sleeves based on calf feature classification.

This study explored the relationship between the design of calf compression sleeves and the comfort of young women in long sitting and standing work environments. By studying the relationship between material elongation and comfort pressure, a prediction model was obtained. The characteristics of the calf models of 94 women were classified, and the influence of different calf characteristics on the pressure distribution and tensile value design was objectively analyzed through simulation tests. The samples were then produced based on the simulation results, and subjective and objective try-on tests and evaluations were carried out. The results show that the difference in the shape of the calf has a certain impact on the pressure distribution and comfort. The predicted value of the theoretical model is in good agreement with the actual test value. The sample can bring a suitable gradient pressure and meet the comfort requirements and safety standards.

Expression of TFRC helps to improve the antineoplastic effect of Ara-C on AML cells through a targeted delivery carrier.

BACKGROUND: Currently, high doses of cytarabine arabinoside (Ara-C)-based combined chemotherapy are commonly used in acute myeloid leukemia (AML) therapy, but severe adverse effects and poor suppression effects in leukemia cells limit the clinical therapeutic efficiency of Ara-C-based chemotherapy due to a lack of targeting selectivity. To improve the therapeutic effect of Ara-C in AML, here, since we confirmed that transferrin receptor 1 (TFRC) expression in AML cells was constant, we generated Ara-C@HFn by encapsulating free Ara-C into self-assembled heavy ferritin chain (HFn, the ligand of TFRC) nanocages. RESULTS: The analysis of clinically relevant data suggested that the high expression levels of TFRC from AML cells would not decrease significantly after treatment with Ara-C. Ara-C@HFn can be efficiently internalized by leukemia cells, showing stronger cytotoxic effects in vitro and reducing the burden of leukemia in AML mice more effectively in vivo than free Ara-C. Ara-C@HFn treatment showed no acute toxicity in visceral organs of mice. Moreover, the analysis of clinically relevant data also suggested that there are several drugs (such as tamibarotene and ABT199) that would not cause significant expression down-regulation of TFRC in AML cells (after treatment). CONCLUSION: The above results suggested that TFRC can be used as a constant and effective target for drug targeting delivery of AML cells. Thus Ara-C@HFn treatment can become a safe and efficient strategy for AML therapy by specifically delivering Ara-C to AML cells. Besides, the HFn nanocages are promising for improving antineoplastic effect of other AML-related therapy drugs that do not cause downregulated expression of TFRC in AML cells.

Integrated mass spectrometry imaging and metabolomics reveals sublethal effects of indoxacarb on the red fire ant Solenopsis invicta.

BACKGROUND: Indoxacarb, representing an efficient insecticide, is normally made into a bait to spread the poison among red fire ants so that it can be widely applied in the prevention and control of Solenopsis invicta. However, the potential toxicity mechanism of S. invicta in response to indoxacarb remains to be explored. In this study, we integrated mass spectrometry imaging (MSI) and untargeted metabolomics methods to reveal disturbed metabolic expression levels and spatial distribution within the whole-body tissue of S. invicta treated with indoxacarb. RESULTS: Metabolomics results showed a significantly altered level of metabolites after indoxacarb treatment, such as carbohydrates, amino acids and pyrimidine and derivatives. Additionally, the spatial distribution and regulation of several crucial metabolites resulting from the metabolic pathway and lipids can be visualized using label-free MSI methods. Specifically, xylitol, aspartate, and uracil were distributed throughout the whole body of S. invicta, while sucrose-6'-phosphate and glycerol were mainly distributed in the abdomen of S. invicta, and thymine was distributed in the head and chest of S. invicta. Taken together, the integrated MSI and metabolomics results indicated that the toxicity mechanism of indoxacarb in S. invicta is closely associated with the disturbance in several key metabolic pathways, such as pyrimidine metabolism, aspartate metabolism, pentose and glucuronate interconversions, and inhibited energy synthesis. CONCLUSION: Collectively, these findings provide a new perspective for the understanding of toxicity assessment between targeted organisms S. invicta and pesticides. © 2023 Society of Chemical Industry.

Metabolomics and mass spectrometry imaging reveal the chronic toxicity of indoxacarb to adult zebrafish (Danio rerio) livers.

Indoxacarb is a widely used insecticide in the prevention and control of agricultural pests, whereas its negative effects on non-target organisms remain largely unclear. Herein, we demonstrated the integrated metabolomics and mass spectrometry imaging (MSI) methods to investigate the chronic exposure toxicity of indoxacarb at environmentally relevant concentrations in adult zebrafish (Danio rerio) liver. Results showed that movement behaviors of zebrafish can be affected and catalase (CAT), glutamic oxalacetic transaminase (GOT), and glutamic pyruvic transaminase (GPT) activities were significantly increased after indoxacarb exposure for 28 days. Pathological analysis of zebrafish livers also showed that cavitation and pathological reactions occur. Metabolomics results indicated that metabolic pathways of zebrafish liver could be significantly affected by indoxacarb, such as tricarboxylic acid (TCA) cycle and various amino acid metabolisms. MSI results revealed the spatial differentiation of crucial metabolites involved in these metabolic pathways within zebrafish liver. Taken together, these integrated MSI and metabolomics results revealed that the toxicity of indoxacarb arises from metabolic pathways disturbance, which resulted in the decrease of liver detoxification ability. These findings will promote the current understanding of pesticide risks and metabolic disorders in zebrafish liver, which provide new insights into the environmental risk assessment of insecticides on aquatic organisms.

Mechanism of Rotenone Toxicity against Plutella xylostella: New Perspective from a Spatial Metabolomics and Lipidomics Study.

The botanical pesticide rotenone can effectively control target pest Plutella xylostella, yet insights into in situ metabolic regulation of P. xylostella toward rotenone remain limited. Herein, we demonstrated metabolic expression levels and spatial distribution of rotenone-treated P. xylostella using spatial metabolomics and lipidomics. Specifically, rotenone significantly affected purine and amino acid metabolisms, indicating that adenosine monophosphate and inosine were distributed in the whole body of P. xylostella with elevated levels, while guanosine 5'-monophosphate and tryptophan were significantly downregulated. Spatial lipidomics results indicated that rotenone may significantly destroy glycerophospholipids in cell membranes of P. xylostella, inhibit fatty acid biosynthesis, and consume diacylglycerol to enhance fat oxidation. These findings revealed that high toxicity of rotenone toward P. xylostella may be ascribed to negative effects on energy production and amino acid synthesis and damage to cell membranes, providing guidelines for the toxicity mechanism of rotenone on target pests and rational development of botanical pesticide candidates.

High performance printed organic electrochromic devices based on an optimized UV curable solid-state electrolyte.

Manufacturing cost is a major concern for electrochromic device (ECD) applications in smart windows for energy saving and low-carbon economy. Fully printing instead of a vacuum-based chemical vapor deposition (CVD) process is favored for large-scale fabrication of ECDs. To adapt to the screen printing process, a UV curable solid-state electrolyte based on lithium bis(trifluoromethane-sulfonyl) imide (LiTFSI) was specially formulated. It contains poly(ethylene glycol) diacrylate (PEG-DA), LiTFSI, water, and ethyl acetate. The optimized ECDs have achieved a 0.6 s bleaching time at 0.6 V and a 1.4 s coloring time at -0.5 V. The ECDs also exhibited excellent stability, which could endure 100 000 cycles of color switching while still maintaining 35% of transmittance change at a 550 nm wavelength. A demo ECD has been fabricated with a screen printed electrolyte, exhibiting stable switching between the clear state and patterned color state.

Comparative Study on Thermal Response Mechanism of Two Binders during Slow Cook-Off.

The HTPE (hydroxyl-terminated polyether) propellant had a lower ignition temperature (150 °C vs. 240 °C) than the HTPB (hydroxy-terminated polybutadiene) propellant in the slow cook-off test. The reactions of the two propellants were combustion and explosion, respectively. A series of experiments including the changes of colors and the intensity of infrared characteristic peaks were designed to characterize the differences in the thermal response mechanisms of the HTPB and HTPE binder systems. As a solid phase filler to accidental ignition, the weight loss and microscopic morphology of AP (30~230 °C) were observed by TG and SEM. The defects of the propellant caused by the cook-off were quantitatively analyzed by the box counting method. Above 120 °C, the HTPE propellant began to melt and disperse in the holes, filling the cracks, which generated during the decomposition of AP at a low temperature. Melting products were called the "high-temperature self-repair body". A series of analyses proved that the different thermal responses of the two binders were the main cause of the slow cook-off results, which were likewise verified in the propellant mechanical properties and gel fraction test. From the microscopic point of view, the mechanism of HTPE's slow cook-off performance superior to HTPB was revealed in this article.

Sample preparation optimization of insects and zebrafish for whole-body mass spectrometry imaging.

Appropriate sample preparation is one of the most critical steps in mass spectrometry imaging (MSI), which is closely associated with reproducible and reliable images. Despite that model insects and organisms have been widely used in various research fields, including toxicology, drug discovery, disease models, and neurobiology, a systematic investigation on sample preparation optimization for MSI analysis has been relatively rare. Unlike mammalian tissues with satisfactory homogeneity, freezing sectioning of the whole body of insects is still challenging because some insect tissues are hard on the outside and soft on the inside, especially for some small and fragile insects. Herein, we systematically investigated the sample preparation conditions of various insects and model organisms, including honeybees (Apis cerana), oriental fruit flies (Bactrocera dorsalis), zebrafish (Danio rerio), fall armyworms (Spodoptera frugiperda), and diamondback moths (Plutella xylostella), for MSI. Three cutting temperatures, four embedding agents, and seven thicknesses were comprehensively investigated to achieve optimal sample preparation protocols for MSI analysis. The results presented herein indicated that the optimal cutting temperature and embedding agent were -20 °C and gelatin, respectively, providing better tissue integrity and less mass spectral interference. However, the optimal thickness for different organisms can vary with each individual. Using this optimized protocol, we exploited the potential of MSI for visualizing the tissue-specific distribution of endogenous lipids in four insects and zebrafish. Taken together, this work provides guidelines for the optimized sample preparation of insects and model organisms, facilitating the expansion of the potential of MSI in the life sciences and environmental sciences.

Nitrogen-rich energetic polymer powered aluminum particles with enhanced reactivity and energy content.

Aluminum particles are of significant interest in enhancing the energy release performance of explosives. One of the major impediments to their use is that Al2O3 shell significantly decreases overall performance. To address this issue, we investigate creating aluminum particles with a glycidyl azide polymer (GAP) coating to improve their reactivity while retaining their energy content. We found that the aluminum particles were coated with a GAP layer of thickness around 8.5 nm. The coated aluminum particles were compared to non-coated powder by the corresponding reactivity parameters obtained from simultaneous differential scanning calorimetry, thermal gravimetric analysis, coupled with mass spectral and infrared spectral analyses. Besides, the comparison on the energy content was also conducted based on P-t tests and a laser-induced air shock from energetic materials (LASEM) technique. It was found that GAP shifted the oxidation onset of aluminum particles to a lower temperature by ~ 10 °C. Besides, the oxidation activation energy of aluminum particles was also reduced by ~ 15 kJ mol-1. In return, aluminum particles reduced the activation energy of the second stage decomposition of the GAP by 276 kJ mol-1. And due to the synergistic effect between aluminum and GAP, the decomposition products of GAP were prone to be oxycarbide species rather than carbonitride species. In addition, the P-t test showed the peak pressure and pressurization rate of GAP coated aluminum particles were separately 1.4 times and 1.9 times as large as those of non-coated aluminum particles. Furthermore, the LASEM experiment suggested the shock wave velocity of the GAP coated aluminum particles was larger than that of non-coated aluminum particles, and the largest velocity difference for them could be 0.6 km s-1. This study suggests after coating by GAP, the aluminum particles possess enhanced reaction performance, which shows potential application value in the fields of aluminized explosives and other energetic fields.

Stereoselective toxicity mechanism of neonicotinoid dinotefuran in honeybees: New perspective from a spatial metabolomics study.

Development of stereoisomeric neonicotinoid pesticides with lower toxicity is key to preventing global population declines of honeybees, whereas little is known about the in situ metabolic regulation of honeybees in response to stereoisomeric pesticides. Herein, we demonstrate an integrated mass spectrometry imaging (MSI) and untargeted metabolomics method to disclose disturbed metabolic expression levels and spatial differentiation in honeybees (Apis cerana) associated with stereoisomeric dinotefuran. This method affords a metabolic network mapping capability regarding a wide range of metabolites involved in multiple metabolic pathways in honeybees. Metabolomics results indicate more metabolic pathways of honeybees can be significantly affected by S-(+)-dinotefuran than R-(-)-dinotefuran, such as tricarboxylic acid (TCA) cycle, glyoxylate and dicarboxylate metabolism, and various amino acid metabolisms. MSI results demonstrate the cross-regulation and spatial differentiation of crucial metabolites involved in the TCA cycle, purine, glycolysis, and amino acid metabolisms within honeybees. Taken together, the integrated MSI and metabolomics results indicated the higher toxicity of S-(+)-dinotefuran arises from metabolic pathway disturbance and its inhibitory role in the energy metabolism, resulting in significantly reduced degradation rates of detoxification mechanisms. From the view of spatial metabolomics, our findings provide novel perspectives for the development and applications of pure chiral agrochemicals.

Fabrication of Energetic Composites with 91% Solid Content by 3D Direct Writing.

Direct writing is a rapidly developing manufacturing technology that is convenient, adaptable and automated. It has been used in energetic composites to manufacture complex structures, improve product safety, and reduce waste. This work is aimed at probing the formability properties and combustion performances of aluminum/ammonium perchlorate with a high solid content for direct writing fabrication. Four kinds of samples with different solid content were successfully printed by adjusting printing parameters and inks formulas with excellent rheological behavior and combustion properties. A high solid content of 91% was manufactured and facile processed into complex structures. Micromorphology, rheology, density, burning rate, heat of combustion and combustion performance were evaluated to characterized four kinds of samples. As the solid content increases, the density, burning rate and heat of combustion are greatly enhanced. Based on 3D direct writing technology, complex energetic 3D structures with 91% solid content are shaped easier and more flexibly than in traditional manufacturing process, which provides a novel way for the manufacture of complicated structures of energetic components.

Spatiotemporal Visualization of Insecticides and Fungicides within Fruits and Vegetables Using Gold Nanoparticle-Immersed Paper Imprinting Mass Spectrometry Imaging.

Food safety issues caused by pesticide residue have exerted far-reaching impacts on human daily life, yet the available detection methods normally focus on surface residue rather than pesticide penetration to the internal area of foods. Herein, we demonstrated gold nanoparticle (AuNP)-immersed paper imprinting mass spectrometry imaging (MSI) for monitoring pesticide migration behaviors in various fruits and vegetables (i.e., apple, cucumber, pepper, plum, carrot, and strawberry). By manually stamping food tissues onto AuNP-immersed paper, this method affords the spatiotemporal visualization of insecticides and fungicides within fruits and vegetables, avoiding tedious and time-consuming sample preparation. Using the established MSI platform, we can track the migration of insecticides and fungicides into the inner region of foods. The results revealed that both the octanol-water partition coefficient of pesticides and water content of garden stuffs could influence the discrepancy in the migration speed of pesticides into food kernels. Taken together, this nanopaper imprinting MSI is poised to be a powerful tool because of its simplicity, rapidity, and easy operation, offering the potential to facilitate further applications in food analysis. Moreover, new perspectives are given to provide guidelines for the rational design of novel pesticide candidates, reducing the risk of food safety issues caused by pesticide residue.

Insights into the degradation and toxicity difference mechanism of neonicotinoid pesticides in honeybees by mass spectrometry imaging.

Honeybees are essential for the pollination of a wide variety of crops and flowering plants, whereas they are confronting decline around the world due to the overuse of pesticides, especially neonicotinoids. The mechanism behind the negative impacts of neonicotinoids on honeybees has attracted considerable interest, yet it remains unknown due to the limited insights into the spatiotemporal distribution of pesticides in honeybees. Herein, we demonstrated the use of matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) for the spatiotemporal visualization of neonicotinoids, such as N-nitroguanidine (dinotefuran) and N-cyanoamidine (acetamiprid) compounds, administered by oral application or direct contact, in the whole-body section of honeybees. The MSI results revealed that both dinotefuran and acetamiprid can quickly penetrate various biological barriers and distribute within the whole-body section of honeybees, but acetamiprid can be degraded much faster than dinotefuran. The degradation rate of acetamiprid is significantly decreased when piperonyl butoxide (PBO) is applied, whereas that of dinotefuran remains almost unchanged. These two factors might contribute to the fact that dinotefuran affords a higher toxicity to honeybees than acetamiprid. Moreover, the toxicity and degradation rate of acetamiprid can be affected by co-application with tebuconazole. Taken together, the results presented here indicate that the discrepant toxicity between dinotefuran and acetamiprid does not lie in the difference in their penetration of various biological barriers of honeybees, but in the degradation rate of neonicotinoid pesticides within honeybee tissues. Moreover, new perspectives are given to better understand the causes of the current decline in honeybee populations posed by insecticides, providing guidelines for the precise use of conventional agrochemicals and the rational design of novel pesticide candidates.

Rapid structural discrimination of IgG antibodies by multicharge-state collision-induced unfolding.

Immunoglobulin G (IgG) antibodies are an important class of biotherapeutics that target various diseases, such as cancers, neurodegenerative disorders, and autoimmune diseases, yet rapid discrimination of IgG antibodies remains a great challenge due to heterogeneity, flexibility, and large size. Herein, we demonstrate a simplified multicharge-state collision-induced unfolding (CIU) method for rapid differentiation of four IgG isotypes that differ in terms of the numbers and patterns of disulfide bonds, bypassing tedious single charge-state selection in advance. The results presented herein reveal that gas-phase unfolding behaviors have a strong dependence on charge states outside IgG surfaces; therefore, multicharge-state CIU analysis of IgG subtypes could offer a great opportunity to gain deeper insights into their gas-phase structural differentiation. The full discrimination of IgG antibody isoforms that possess different disulfide bond numbers and even subtle disulfide bonding patterns can be achieved based on their charge-dependent gas-phase unfolding behaviors and root-mean square deviation in CIU difference spectra. Taken together, the incorporation of all charge states observed in a native ion mobility-mass spectrometry (IM-MS) experiment to CIU analysis could make this strategy sensitive to more subtle structural discrepancies, facilitating the rapid discrimination and evaluation of innovative structurally similar biotherapeutic candidates with unexplored functions.

Discrimination of isomeric monosaccharide derivatives using collision-induced fingerprinting coupled to ion mobility mass spectrometry.

Because of the isomeric heterogeneity that is ubiquitous in analytical science, a formidable analytical challenge is to fully discriminate multiple isomers, especially those candidate isomers with various biological functions. Ion mobility mass spectrometry (IM-MS) has gained impressive advances for gaining molecular conformations, whereas coexisting structurally similar isomers often make unambiguous discrimination impossible due to the limited IM resolution of commercially available instruments. Herein, we demonstrate an energy-resolved collision-induced fingerprint (CIF) method to fully discriminate isomeric monosaccharide derivatives that differ in terms of composition, connectivity and configuration without complex instrument modifications. By simply increasing the collisional energy in the trap cell, the full width at half maximum (FWHM) of IM peaks can be markedly narrowed by at least 2-fold. Given the excellent reproducibility of CIF measurements, the full discrimination of isomers can benefit from their unique feature values and root-mean square deviation (RMSD) in CIF spectra. Moreover, rapid discrimination of each monosaccharide derivate isomer from binary mixtures is demonstrated. This strategy will expand the horizons of IM-MS platform in the rapid differentiation of a wider range of isomers more than monosaccharide derivatives in complex systems, which facilitates the identification and evaluation of innovative isomer candidates with unexplored functions.

Integrin-mediated Signaling via Paxillin-GIT1-PIX Promotes Localized Rac Activation at the Leading Edge and Cell Migration.

Rac activation is precisely regulated temporally and spatially by intracellular signaling pathways in migrating cells to guarantee the formation of specific cell protrusions-lamellipodia at the leading edge. Integrins-mediated adhesions also control the signaling pathway for localized Rac activation in the cells, but very few studies have been addressed in this field. In the study, we aim to focus on how integrin-mediated signaling affects localized Rac activation by reducing the paxillin expression with shRNA targeting paxillin. The results revealed that reduction of the paxillin expression in the cells inhibited the formation of focal adhesions and Rac activation. By using Rac FRET biosensor, Rac activation was localized at the leading edge of the cell, within the lamellipodium. A ternary complex of paxillin-GIT1-PIX could establish the signaling pathway in front of the cells. Thus, we described a mechanism of integrin-mediated signaling for localized Rac activation that upon ligand binding, activated integrin via the signaling pathway paxillin-GIT1-PIX promotes localized Rac activation at the leading edge and cell migration.

SERS-active substrate assembled by Ag NW-embedded porous polystyrene fibers.

Here we demonstrate a novel SERS-active substrate assembled by silver nanowire (Ag NW)-embedded porous polystyrene (PS) fibers. Ag NWs are synthesized through a glycerol-mediated solvothermal method firstly, then electrospun into PS porous fibers. The as-synthesized Ag NWs are embedded in PS fiber and aligned orderly along the axial direction. Porous structure appears in PS fiber due to the phase separation induced by rapid evaporation of solvents. Large amounts of holes not only greatly improve the sample collection efficiency of the SERS-active substrate, but also significantly facilitate the adsorption of target molecules on the surface of Ag NWs, thus increasing the probability of enhancement of target molecules. In addition, compared with polyvinyl alcohol (PVA) and polyvinyl pyrrolidone (PVP), PS has better solvent resistance. The detection limit of 4-aminothiophenol (4-ATP) on our fabricated electrospun fiber mats is 10-7 M, and the electrospun fiber mats showed good reproducibility of SERS signal detection. This study proposes a feasible strategy for the large-scale preparation of flexible SERS-active substrate assembled by Ag NW-embedded porous PS fibers. The produced flexible SERS substrates may have potential application in wearable sensors for the trace detection of chemical and biological molecules.

Newly Discovered Action of HpTx3 from Venom of Heteropoda venatoria on Nav1.7 and Its Pharmacological Implications in Analgesia.

It has been reported that Heteropodatoxin3 (HpTx3), a peptidic neurotoxin purified from the venom of the spider species Heteropoda venatoria, could inhibit Kv4.2 channels. Our present study newly found that HpTx3 also has potent and selective inhibitory action on Nav1.7, with an IC50 of 135.61 ± 12.98 nM. Without effect on the current-voltage (I-V) relationship of Nav1.7, HpTx3 made minor alternation in the voltage-dependence of activation and steady-state inactivation of Nav1.7 (4.15 mV and 7.29 mV, respectively) by interacting with the extracellular S3-S4 loop (S3b-S4 sequence) in domain II and the domain IV of the Nav channel subtype, showing the characteristics of both pore blocker and gate modifier toxin. During the interaction of HpTx3 with the S3b-S4 sequence of Nav1.7, the amino acid residue D in the sequence played a key role. When administered intraperitoneally or intramuscularly, HpTx3 displayed potent analgesic activity in a dose-dependent manner in different mouse pain models induced by formalin, acetic acid, complete Freund's adjuvant, hot plate, or spared nerve injury, demonstrating that acute, inflammatory, and neuropathic pains were all effectively inhibited by the toxin. In most cases HpTx3 at doses of ≥ 1mg/kg could produce the analgesic effect comparable to that of 1 mg/kg morphine. These results suggest that HpTx3 not only can be used as a molecular probe to investigate ion channel function and pain mechanism, but also has potential in the development of the drugs that treat the Nav1.7 channel-related pain.

Replication and virulence in pigs of the first African swine fever virus isolated in China.

African swine fever (ASF) entered China in August 2018 and rapidly spread across the entire country, severely threatening the Chinese domestic pig population, which accounts for more than 50% of the pig population worldwide. In this study, an ASFV isolate, Pig/Heilongjiang/2018 (Pig/HLJ/18), was isolated in primary porcine alveolar macrophages (PAMs) from a pig sample from an ASF outbreak farm. The isolate was characterized by using the haemadsorption (HAD) test, Western blotting and immunofluorescence, and electronic microscopy. Phylogenetic analysis of the viral p72 gene revealed that Pig/HLJ/18 belongs to Genotype II. Infectious titres of virus propagated in primary PAMs and pig marrow macrophages were as high as 107.2 HAD50/ml. Specific-pathogen-free pigs intramuscularly inoculated with different virus dosages at 103.5-106.5 HAD50 showed acute disease with fever and haemorrhagic signs. The incubation periods were 3-5 days for virus-inoculated pigs and 9 days for contact pigs. All virus-inoculated pigs died between 6-9 days post-inoculation (p.i.), and the contact pigs died between 13-14 days post-contact (p.c.). Viremia started on day 2 p.i. in inoculated pigs and on day 9 p.c. in contact pigs. Viral genomic DNA started to be detected from oral and rectal swab samples on 2-5 days p.i. in virus-inoculated pigs, and 6-10 days p.c. in contact pigs. These results indicate that Pig/HLJ/18 is highly virulent and transmissible in domestic pigs. Our study demonstrates the threat of ASFV and emphasizes the need to control and eradicate ASF in China.