Development of a LAMP assay for the rapid visual detection of the emerging tick-borne Songling virus.

BACKGROUND: Songling virus (SGLV) within the genus Orthonairovirus, family Nairoviridae, is an emerging tick-borne virus associated with human febrile illness. However, no rapid detection method for SGLV has been established. METHODS: In this study, four primer sets targeting the nucleocapsid protein gene of SGLV were designed for use in the LAMP assay and evaluated to identify the optimal primer set. Recombinant plasmids were constructed and utilized for assessing the sensitivity of the assay. Tacheng tick virus 1 (TcTV-1)-, Beiji nairovirus (BJNV)-, Yezo virus (YEZV)-, severe fever with thrombocytopenia syndrome virus (SFTSV)-, and tick-borne encephalitis virus (TBEV)-positive tick samples were utilized to assess the specificity. Field-collected ticks were also evaluated as biological specimens to validate the assay. RESULTS: A SGLV-specific LAMP assay was established with a detection limit of 1 × 10-2 copies/µl and could be visually confirmed by a color change from purple to blue in SGLV-positive samples. No cross-reactivity was observed in the detection of TcTV-1, BJNV, YEZV, SFTSV, and TBEV using the LAMP assay. In addition to the detection of the same seven high-copy numbers of SGLV as the SYBR Green quantitative RT-PCR assay within a reduced timeframe, the developed LAMP method also effectively identified an additional sample with a low copy number in the field-collected tick samples. CONCLUSIONS: We successfully developed a sensitive, specific, and cost-effective visual method for the rapid detection of SGLV using the LAMP assay, which can be applied in pathogenesis and epidemiological surveillance studies of SGLV.

Temperature Compensation for MEMS Accelerometer Based on a Fusion Algorithm.

This study proposes a fusion algorithm based on forward linear prediction (FLP) and particle swarm optimization-back propagation (PSO-BP) to compensate for the temperature drift. Firstly, the accelerometer signal is broken down into several intrinsic mode functions (IMFs) using variational modal decomposition (VMD); then, according to the FE algorithm, the IMF signal is separated into mixed components, temperature drift, and pure noise. After that, the mixed noise is denoised by FLP, and PSO-BP is employed to create a model for temperature adjustment. Finally, the processed mixed noise and the processed IMFs are rebuilt to obtain the enhanced output signal. To confirm that the suggested strategy works, temperature experiments are conducted. After the output signal is processed by the VMD-FE-FLP-PSO-BP algorithm, the acceleration random walk has been improved by 23%, the zero deviation has been enhanced by 24%, and the temperature coefficient has been enhanced by 92%, compared with the original signal.

MEMS Gyroscope Temperature Compensation Based on Improved Complete Ensemble Empirical Mode Decomposition and Optimized Extreme Learning Machine.

Herein, we investigate the temperature compensation for a dual-mass MEMS gyroscope. After introducing and simulating the dual-mass MEMS gyroscope's working modes, we propose a hybrid algorithm for temperature compensation relying on improved complete ensemble empirical mode decomposition with adaptive noise (ICEEMDAN), sample entropy, time-frequency peak filtering, non-dominated sorting genetic algorithm-II (NSGA II) and extreme learning machine. Firstly, we use ICEEMDAN to decompose the gyroscope's output signal, and then we use sample entropy to classify the decomposed signals. For noise segments and mixed segments with different levels of noise, we use time-frequency peak filtering with different window lengths to achieve a trade-off between noise removal and signal retention. For the feature segment with temperature drift, we build a compensation model using extreme learning machine. To improve the compensation accuracy, NSGA II is used to optimize extreme learning machine, with the prediction error and the 2-norm of the output-layer connection weight as the optimization objectives. Enormous simulation experiments prove the excellent performance of our proposed scheme, which can achieve trade-offs in signal decomposition, classification, denoising and compensation. The improvement in the compensated gyroscope's output signal is analyzed based on Allen variance; its angle random walk is decreased from 0.531076°/h/√Hz to 6.65894 × 10-3°/h/√Hz and its bias stability is decreased from 32.7364°/h to 0.259247°/h.

First detection of Candidatus Rickettsia tarasevichiae in Hyalomma marginatum ticks.

Ticks are important vectors of zoonotic diseases and play a major role in the circulation and transmission of many rickettsial species. The aim of this study was to investigate the carriage of Candidatus Rickettsia tarasevichiae (CRT) in a total of 1168 ticks collected in Inner Mongolia to elucidate the potential public health risk of this pathogen, provide a basis for infectious disease prevention, control and prediction and contribute diagnostic ideas for clinical diseases that present with fever in populations exposed to ticks. A total of four tick species, Haemaphysalis concinna (n = 21), Dermacentor nuttalli (n = 122), Hyalomma marginatum (n = 148), and Ixodes persulcatus (n = 877), were collected at nine sampling sites in Inner Mongolia, China, and identified by morphological and molecular biological methods. Reverse transcription PCR targeting the 16S ribosomal RNA (rrs), gltA, groEL, ompB and Sca4 genes was used to detect CRT DNA. Sequencing was used for pathogen species confirmation. The molecular epidemiological analysis showed that three species of ticks were infected with CRT, and the overall positive rate was as high as 42%. The positive rate of I. persulcatus collected in Hinggan League city was up to 96%, and that of I. persulcatus collected in Hulun Buir city was 50%. The pool positive rates of D. nuttalli and H. marginatum collected in Bayan Nur city and H. concinna collected in Hulun Buir city were 0%, 28% and 40%, respectively. This study revealed the high prevalence of CRT infection in ticks from Inner Mongolia and the first confirmation of CRT detected in H. marginatum in China. The wide host range and high infection rate in Inner Mongolia may dramatically increase the exposure of CRT to humans and other vertebrates. The role of H. marginatum in the transmission of rickettsiosis and its potential risk to public health should be further considered.

Microbial diversity of ticks and a novel typhus group Rickettsia species (Rickettsiales bacterium Ac37b) in Inner Mongolia, China.

Ticks can carry multiple pathogens, and Inner Mongolia's animal husbandry provides excellent environmental conditions for ticks. This study characterized the microbiome of ticks from different geographical locations in Inner Mongolia; 905 Dermacentor nuttalli and 36 Ixodes persulcatus were collected from sheep in three main pasture areas and from bushes within the forested area. Mixed DNA samples were prepared from three specimens from each region and tick species. Microbial diversity was analyzed by 16S rRNA sequencing, and α and ß diversity were determined. The predominant bacterial genera were Rickettsia (54.60%), including Rickettsiales bacterium Ac37b (19.33%) and other Rickettsia (35.27%), Arsenophonus (11.21%), Candidatus Lariskella (10.84%), and Acinetobacter (7.17%). Rickettsia bellii was identified in I. persulcatus, while Rickettsiales bacterium Ac37b was found in D. nuttalli from Ordos and Chifeng. Potential Rickettsia and Anaplasma coinfections were observed in the Ordos region. Tick microbial diversity analysis in Inner Mongolia suggests that sheep at the sampling sites were exposed to multiple pathogens.

Title: Diversité microbienne des tiques et nouvelle espèce de Rickettsia du groupe du typhus (bactérie Rickettsiales Ac37b) en Mongolie intérieure, Chine. Abstract: Les tiques peuvent être porteuses de plusieurs agents pathogènes et l'élevage en Mongolie intérieure offre d'excellentes conditions environnementales pour les tiques. Cette étude a caractérisé le microbiome des tiques de différentes zones géographiques de Mongolie intérieure; 905 Dermacentor nuttalli et 36 Ixodes persulcatus ont été collectés sur des moutons dans trois principales zones de pâturage et dans des buissons de la zone forestière. Des échantillons d'ADN mixtes ont été préparés à partir de trois spécimens de chaque région et espèce de tique. La diversité microbienne a été analysée par séquençage de l'ARNr 16S et la diversité α et ß a été déterminée. Les genres bactériens prédominants étaient les Rickettsia (54,60 %), dont la bactérie Rickettsiales Ac37b (19,33 %) et d'autres Rickettsia (35,27 %), Arsenophonus (11,21 %), Candidatus Lariskella (10,84 %) et Acinetobacter (7,17 %). Rickettsia bellii a été identifiée chez I. persulcatus, tandis que la bactérie Rickettsiales Ac37b a été trouvée chez D. nuttalli d'Ordos et Chifeng. Des co-infections potentielles à Rickettsia et Anaplasma ont été observées dans la région d'Ordos. L'analyse de la diversité microbienne des tiques en Mongolie intérieure montre que les moutons présents sur les sites d'échantillonnage sont exposés à plusieurs agents pathogènes.

Development and Prospect of Smart Materials and Structures for Aerospace Sensing Systems and Applications.

The rapid development of the aviation industry has put forward higher and higher requirements for material properties, and the research on smart material structure has also received widespread attention. Smart materials (e.g., piezoelectric materials, shape memory materials, and giant magnetostrictive materials) have unique physical properties and excellent integration properties, and they perform well as sensors or actuators in the aviation industry, providing a solid material foundation for various intelligent applications in the aviation industry. As a popular smart material, piezoelectric materials have a large number of application research in structural health monitoring, energy harvest, vibration and noise control, damage control, and other fields. As a unique material with deformation ability, shape memory materials have their own outstanding performance in the field of shape control, low-shock release, vibration control, and impact absorption. At the same time, as a material to assist other structures, it also has important applications in the fields of sealing connection and structural self-healing. Giant magnetostrictive material is a representative advanced material, which has unique application advantages in guided wave monitoring, vibration control, energy harvest, and other directions. In addition, giant magnetostrictive materials themselves have high-resolution output, and there are many studies in the direction of high-precision actuators. Some smart materials are summarized and discussed in the above application directions, aiming at providing a reference for the initial development of follow-up related research.

Dihydroartemisinin-ursodeoxycholic acid conjugate is a potential treatment agent for inflammatory bowel disease.

BACKGROUND: A novel artemisinin derivative, dihydroartemisinin-ursodeoxycholic acid conjugate (4), was found to exhibit strong immunosuppressive activity. Various methods were used to evaluate the immunosuppressive activity and mechanism of action of the compound to explore its potential applications. METHODS: T cell proliferation, mixed lymphocyte reaction (MLR), and Th1/Th17 differentiation assays were used to evaluate the immunosuppressive activity of the compound. Differentially expressed genes from RNA sequencing were analysed with Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, while enriched signalling pathways were further validated by western blotting (WB). In vivo efficacy was validated with delayed-type hypersensitivity (DTH) mouse models and dextran sodium sulphate (DSS)-induced inflammatory bowel disease (IBD) mouse model. RESULTS: Compound 4 inhibited concanavalin A -induced mouse splenic T cell proliferation (IC50 = 15 nM) and anti-CD3/CD28-induced human primary T cell proliferation (IC50 = 30 nM) while also reducing the secretion of hIFN-γ. Compound 4 exhibited similar inhibitory activity in MLR assay. Compound 4 dose-dependently inhibited human Th1/Th17 differentiation. The KEGG pathway enrichment analysis indicated that the genes related to T cell activation signalling pathways PI3K-AKT, MAPK, and NF-κB were significantly enriched. WB confirmed that compound 4 inhibited the AKT/MAPK and NF-κB signalling pathways. Compound 4 dose-dependently inhibited ear and foot pad swelling in DTH mouse models. In the DSS-induced IBD mouse model, compound 4 significantly decreased the disease activity index and colon density, and inhibited splenomegaly of the mice. CONCLUSION: The in vitro and in vivo results indicated that compound 4 has the potential to be developed into an anti-IBD drug.

Identification and genetic diversity analysis of Rickettsia in Dermacentor nuttalli within inner Mongolia, China.

BACKGROUND: The genus Rickettsia contains the lineages spotted fever group (SFG), typhus group (TG), and transitional group (TRG). The spotted fever group Rickettsia (SFGR) is transmitted by ticks. The tick species Dermacentor nuttalli is considered the main vector carrying SFGR in Inner Mongolia. Studying the genetic diversity and population structure of Rickettsia is essential for developing effective control strategies and predicting evolutionary trends of Rickettsia. METHODS: In 2019 we collected 408 D. nuttalli in the Inner Mongolia Autonomous Region, detected the percentage of Rickettsia-positive specimens, and characterized the haplotypes. From the Rickettsia-positive ticks, the gltA and ompA genes were extracted, amplified, and sequenced. RESULTS: Ten haplotypes of the gltA gene and 22 haplotypes of the ompA gene were obtained. The phylogenetic analysis showed that the haplotypes G1-G7 and G9 of the gltA gene cluster with Rickettsia raoultii, while G8 and G10 cluster with Rickettsia sibirica. Haplotypes O1-O15, O18 and O20-O22 of the ompA gene cluster with R. raoultii, while O16 and O19 cluster with R. sibirica. The average haplotype diversity was 0.3 for gltA and 0.7 for ompA. The average nucleotide diversity was greater than 0.05. Neutrality tests were nonsignificant for Tajima's D results and Fu's Fs results. The fixation index values (FST) showed that the degree of genetic differentiation between most sampled populations was small (FST < 0.05), whereas some populations showed a medium (FST > 0.05) or large (FST > 0.15) degree of differentiation. Analysis of molecular variance (AMOVA) revealed that the variation within populations was greater than that between populations. The mismatch analysis of Rickettsia showed double peaks. CONCLUSIONS: We found two Rickettsia spp. (R. raoultii and R. sibirica). The high genetic disparity of Rickettsia allows for easy adaption to different environments. Genetic differentiation between populations is small, and Rickettsia populations do not show a geographically differentiated structure. The high rates of retention and infection of Rickettsia in D. nuttalli together with the animal husbandry exchange in Inner Mongolia gradually led to the harmonization of genetic characteristics of Rickettsia across various regions. Overall, the significant genetic diversity and geographical structure of Rickettsia in D. nuttalli are critical for SFGR control.

Enrichment of heavy metals from spent printed circuit boards by microwave pyrolysis.

This study reports the enrichment behaviors of heavy metals, including copper, tin, lead and zinc, in the process of microwave pyrolysis of spent printed circuit boards (SPCBs). The SPCB had good microwave absorptivity. Under the optimal conditions of microwave power of 700 W, pyrolysis temperature of 400 °C, dwell time of 5 min, N2 gas flow rate of 1.2 L/min, and load mass of 5 g, the yield of pyrolyzed SPCB was 79.16%. The contents of copper, tin, lead, and zinc in the pyrolyzed SPCB were increased to 28.52 wt%, 7.15 wt%, 1.31 wt%, and 1.13 wt%, respectively, with the corresponding retention percentages of 99.98%, 85.89%, 92.59% and 82.06%. The loss of metals was attributed to volatilization of the elements, which was affected by metal discharge due to excitation of electrons in the metals under microwave irradiation. Little copper loss was found because of the difficult reaction between copper and hydrogen bromide and the very high temperature required by the volatilization of copper. Tin, lead and zinc were mainly volatilized in the form of their metal bromides, including SnBr4, ZnBr2, and PbBr2. By controlling the pyrolysis conditions and metal discharge induced in the microwave field, the metals could be effectively enriched for subsequent treatment with high efficiency.

Co-expression of CD24 and Hsp70 as a prognostic biomarker for lung cancer.

Lung cancer is one of the most common malignant neoplasms worldwide. CD24 is a marker of tumor stem cells that plays an important role in tumorigenesis. Hsp70 is an important molecular chaperone. However, the co-expression and interaction of CD24 and Hsp70, as well as the significance for the prognosis of lung cancer are still unclear. The expression levels of CD24 and Hsp70 were detected by immunohistochemistry and their correlation was analyzed. The expression levels of CD24 mRNA and protein were examined using qRT-PCR and western blotting in SPCA1, A549, H1975, and H1650 cell lines. A CD24-overexpressing cell model was established. The interaction between CD24 and Hsp70 was verified by co-immunoprecipitation and western blotting. CD24 and Hsp70 expression were significantly higher in lung cancer tissues than in adjacent tissues (CD24: p=0.008; Hsp70: p<0.001). CD24 protein expression showed a positive correlation with lymph node metastasis, TNM stage, and vascular cancer thrombus. Hsp70 protein expression showed a positive correlation with differentiation, lymph node metastasis, and TNM stage. CD24 and Hsp70 high expression were also correlated with poor survival. The positive co-expression rate of CD24 and Hsp70 in lung cancer tissues was 52.7% (49/93). CD24 and Hsp70 expression in lung cancer were positively correlated (r=0.368, p<0.001), and co-immunoprecipitation was verified that both endogenous and exogenous CD24 co-precipitated with Hsp70 directly or indirectly. When Hsp70 inhibitor VER15508 was added to A549 cells, Hsp70 and CD24 protein expression were significantly decreased. The present study demonstrated that CD24 and Hsp70 were highly expressed in lung cancer tissues, and associated with invasion, metastasis, and poor survival. Hsp70 may regulate CD24 expression. Co-expression of CD24 and Hsp70 may be a prognostic biomarker for lung cancer.

Genetic diversity analysis of Dermacentor nuttalli within Inner Mongolia, China.

BACKGROUND: Ticks (Arthropoda, Ixodida), after mosquitoes, are the second most prevalent vector of infectious diseases. They are responsible for spreading a multitude of pathogens and threatening the health and welfare of animals and human beings. However, given the history of tick-borne pathogen infections in the Inner Mongolia Autonomous Region of China, surprisingly, neither the genetic diversity nor the spatial distribution of haplotypes within ticks has been studied. METHODS: We characterized the haplotype distribution of Dermacentor nuttalli in four main pastoral areas of the Inner Mongolia Autonomous Region, by sampling 109 individuals (recovered from sheep) in April-August 2019. The 16S rRNA gene, cytochrome c oxidase subunit I (COI), and the internal transcribed spacer 2 region (ITS2) were amplified and sequenced from extracted DNA. RESULTS: Twenty-six haplotypes were identified using 16S rRNA sequences, 57 haplotypes were identified with COI sequences, and 75 haplotypes were identified with ITS2 sequences. Among the three genes, total haplotype diversity was greater than 0.7, while total nucleotide diversity was greater than 0.06. Neutrality tests revealed a significantly negative Tajima's D result, while Fu's Fs was not significantly positive. Fixation index values (FST) indicated that the degree of genetic differentiation among some sampled populations was small, while for others it was moderate. Analysis of molecular variance (AMOVA) revealed that the variation within populations was greater than that among populations. The mismatch analysis of D. nuttalli exhibited double peaks. CONCLUSION: The genetic diversity of D. nuttalli populations in our region can likely adapt to different geographical environments, thereby leading to genetic diversity, and creating genetic differentiation among different populations. However, genetic differentiation is cryptic and does not form a pedigree geographical structure.

Synthesis and in vitro antibacterial activity of new aminothiazole-oximepiperidone cephalosporins.

Four new aminothiazole-oximepiperidone cephalosporins (10a-10d) were synthesized, with their in vitro antibacterial activities against hospital isolated Gram-negative bacteria assessed. The results showed that compounds 10a-10d effectively inhibit a variety of Gram-negative bacteria. Compound 10a was the most potent compound, with comparable activity as ceftazidime. The combination of compound 10a and Avibactam was very active against almost all bacteria tested, which including multidrug resistant K. pneumoniae and A. baumannii. Compared to Avycaz, this combination is more potent against ESBL producing K. pneumoniae. Thus, the combination of 10a and Avibactam is of interest for further studies.

Studies on the stereoselective synthesis and immunosuppressive activity of dihydroartemisinin-O-glycoside derivatives.

Eight new dihydroartemisinin-O-glycosides were synthesized with their relative configurations were determined based on NMR spectrum. In vitro immunosuppressive assay showed that 10α-dihydroartemisinin-ß-O-d-mannoside (19a) demonstrate 88% inhibition towards T cells proliferation and 98% reduction in IFN-γ levels in cell media. These results suggest that dihydroartemisinin-O-glycoside as a potential lead for further in vivo evaluation.

The Combination of CD147 and MMP-9 Serum Levels Is Identified as Novel Chemotherapy Response Markers of Advanced Non-Small-Cell Lung Cancer.

To evaluate the correlation between the changes in serum concentrations of cluster of differentiation-147 (scCD147) and chemotherapy outcome in patients with NSCLC and evaluate the combination of scCD147 with serum matrix metalloproteinase-9 (scMMP-9) levels in the prediction of chemotherapy response, eighty-two patients with advanced LC were enrolled. Newly diagnosed cases were treated with platinum-based chemotherapy. We measured scCD147 protein levels in LC cases by ELISA and used receiver operating characteristic (ROC) curves to analyze the results. Four time points were chosen to examine the association between the changes in scCD147 and chemotherapy outcome: before chemotherapy and 21 days after the start of the first, second, and fourth chemotherapy cycles. We assessed the combination of scCD147 and scMMP-9 serum levels in predicting the chemotherapy response. scCD147 was higher in LC cases than that in healthy volunteers (HVs). scCD147 was associated with distant metastases and TNM stage. scCD147 and scMMP-9 appeared to be independent predictive factors for chemotherapy outcomes after the first and second chemotherapy cycles for patients with NSCLC. Multivariable analysis also demonstrated that variations in scCD147 and scMMP-9 could be independent factors for monitoring chemotherapy outcome for patients with NSCLC. Furthermore, when scCD147 and scMMP-9 are combined into a new risk model, it has a markedly better prediction of chemotherapy outcomes than each protein alone. scCD147 and MMP-9 are potential predictive biomarkers for efficacy, and their combination significantly improves the predictive power for chemotherapy response in patients with NSCLC.

High pressure-assisted vacuum-freeze drying: A novel, efficient way to accelerate moisture migration in shrimp processing.

High pressure processing (HPP), as nonthermal processing technology, has the potential to increase the drying rate due to its improvement of heat and mass exchange in different processes. In this study, the moisture migration in shrimps during HPP-vacuum-freeze drying (HPP-VFD) processes has been monitored by using low-field nuclear magnetic resonance and magnetic resonance image (MRI) in comparison with hot air-drying and VFD. Based on the T2 relaxation spectra, three water fractions corresponding to bound water (hydrogen-bonded water), immobile water (water trapped by organization structure or cell member), and free water were observed. For group B, with increasing drying time (4 to 22 hr), the transverse relaxation times of T21 , T22 , and T23 were significantly decreased (76.79%, 57.78%, and 40.9%) (P < 0.05). The content of immobile water (A22 ) and free water (A23 ) decreased (81.55% and 89.07%), whereas the bound water (A21 ) increased (7.26%). In comparison with group B, the T21 , T22 , and T23 of group C showed greater decrease (83.12%, 87.12%, and 89.57% for group C) so that HPP pretreatment could shorten the relaxation time. MRI analysis further proved that HPP-VFD drying has improved drying efficiency, and moisture migration was from the exterior to the interior part with increasing drying time. SEM analysis demonstrated that no significant damage of muscle fibers with narrower gaps was observed for groups B and C. Overall, HPP, as a pretreatment technology, could accelerate the moisture migration and improve the drying efficiency of VFD process for shrimp. PRACTICAL APPLICATION: High pressure processing (HPP) is now well known as a nonthermal processing technology and becoming increasingly acknowledged. However, there is limited information about its application in shrimp-drying process and the moisture dynamic of shrimp subjected to high pressure processing-assisted vacuum-freeze drying. This study could provide valuable information regarding the moisture status and migration in HPP-VFD shrimp monitored by LF-NMR and MRI methods. The results showed that HPP processing at 550 MPa for 10 min can be used as an interesting method for drying pretreatment, increasing its drying rate and consequently reducing its process time, and it demonstrated that the methods used in this study had good correlation coefficient with physicochemical properties of shrimp, which may be real-time and nondestructive monitoring methods for shrimp-drying process.

Impact of High Hydrostatic Pressure on the Shelling Efficacy, Physicochemical Properties, and Microstructure of Fresh Razor Clam (Sinonovacula constricta).

The effects of high hydrostatic pressure (HHP) treatments (200, 300, and 400 MPa for 1, 3, 5 and 10 min) on the shelling efficacy (the rate of shelling, the rate of integrity and yield of razor clam meat) and the physicochemical (drip loss, water-holding capacity, pH, conductivity, lipid oxidation, Ca2+ -ATPase activity, myofibrillar protein content), microbiological (total viable counts) and microstructural properties of fresh razor clam (Sinonovacula constricta) were investigated. HHP treatments significantly (P < 0.05) increased shelling efficiency, water-holding capacity, pH, conductivity, and lipid oxidation, and HHP-treated razor clam showed lower levels of microorganisms and drip loss than untreated razor clam. Levels of thiobarbituric acid reacting substances (TBA) in HHP-treated razor clam were greatly increased (up to 0.93 ± 0.09 mg MDA/kg at 400 MPa for 10 min) which was caused by the formation of hydroperoxides during HHP treatment. All HHP treatments were found to have adverse effects on the activity of Ca2+ -ATPase and the content of myofibrillar protein (MP), which might be due to the substantial damage to the tertiary structure of proteins at high pressure. Moreover, scanning electron microscopy (SEM) revealed the compaction of the muscle fibers and a decrease in the extracellular space with increasing pressure and holding time. This phenomenon was mainly correlated with the compaction of muscle fibers and denaturation, aggregation, and gelation of muscle protein triggered by high pressure. In general, HHP could be applied as a safe and effective nonthermal technology to produce high-quality shelled razor clam. PRACTICAL APPLICATION: High hydrostatic pressure (HHP) is now well known as a nonthermal processing technology and becoming increasingly acknowledged. However, it has not been widely applied to shell seafood due to its uncertain influence on its quality and shelling property. This study could provide valuable information regarding the shelling efficacy, physicochemical properties, and microstructure of razor clam treated by HHP. And it demonstrated that HHP showed a positive impact on quality of razor clam treated by HHP.

A novel expression vector for the secretion of abaecin in Bacillus subtilis

ABSTRACT This study aimed to describe a Bacillus subtilis expression system based on genetically modified B. subtilis. Abaecin, an antimicrobial peptide obtained from Apis mellifera, can enhance the effect of pore-forming peptides from other species on the inhibition of bacterial growth. For the exogenous expression, the abaecin gene was fused with a tobacco etch virus protease cleavage site, a promoter Pglv, and a mature beta-glucanase signal peptide. Also, a B. subtilis expression system was constructed. The recombinant abaecin gene was expressed and purified as a recombinant protein in the culture supernatant. The purified abaecin did not inhibit the growth of Escherichia coli strain K88. Cecropin A and hymenoptaecin exhibited potent bactericidal activities at concentrations of 1 and 1.5 µM. Combinatorial assays revealed that cecropin A and hymenoptaecin had sublethal concentrations of 0.3 and 0.5 µM. This potentiating functional interaction represents a promising therapeutic strategy. It provides an opportunity to address the rising threat of multidrug-resistant pathogens that are recalcitrant to conventional antibiotics.

A novel expression vector for the secretion of abaecin in Bacillus subtilis.

This study aimed to describe a Bacillus subtilis expression system based on genetically modified B. subtilis. Abaecin, an antimicrobial peptide obtained from Apis mellifera, can enhance the effect of pore-forming peptides from other species on the inhibition of bacterial growth. For the exogenous expression, the abaecin gene was fused with a tobacco etch virus protease cleavage site, a promoter Pglv, and a mature beta-glucanase signal peptide. Also, a B. subtilis expression system was constructed. The recombinant abaecin gene was expressed and purified as a recombinant protein in the culture supernatant. The purified abaecin did not inhibit the growth of Escherichia coli strain K88. Cecropin A and hymenoptaecin exhibited potent bactericidal activities at concentrations of 1 and 1.5µM. Combinatorial assays revealed that cecropin A and hymenoptaecin had sublethal concentrations of 0.3 and 0.5µM. This potentiating functional interaction represents a promising therapeutic strategy. It provides an opportunity to address the rising threat of multidrug-resistant pathogens that are recalcitrant to conventional antibiotics.

Mini-genome rescue of Crimean-Congo hemorrhagic fever virus and research into the evolutionary patterns of its untranslated regions.

Crimean-Congo hemorrhagic fever virus (CCHFV) is a member of genus Nairovirus, family Bunyaviridae, which are distributed widely in Africa, Europe and Asia with several genotypes. As a BSL-4 level pathogen, the requirement of high-level biosafety facilities severely constrains researches on live virus manipulation. In this study, we developed a helper-virus-independent mini-genome rescue system for the Chinese YL04057 strain. Based on the enhanced green fluorescent protein (EGFP)-derived mini-genome plasmids, this polymerase I driven system permits easy observation and quantification. Unlike previous report, gradually reduced levels of activity of the CCHFV L, M and S untranslated regions (UTRs) were observed in our system. We also demonstrated that the UTRs at both ends were indispensable for mini-genome background expression. In addition, we phylogentically analyzed all six UTRs of CCHFV and showed that L-UTRs were clustered together approximately corresponding to their original geographical continents. The UTRs of M segment showed a similar branch structure to its open reading frames (ORFs), and nearly an identical tree was generated with 5' UTRs of S segment compared with its ORFs. However, the 3' UTRs of S segment formed new divergent groups. Compatibility tests of YL04057 strain nucleocapsid protein and L protein expression plasmids with Nigerian strain IbAr10200 mini-genomes revealed lower compatibility of L-UTRs without an obvious effect on M-UTRs. Moreover, we demonstrated that the L-UTRs could tolerate certain nucleotide mutations. This system may provide a foundation for future studies of the viral replication cycle, pathogenic mechanisms and evolutionary patterns of CCHFV.

An improved system for the evaluation of antiviral compounds against herpes simplex virus type 2.

Infection with herpes simplex virus type 2 (HSV-2) can result in lesions in reproductive organs, along with long-term latency. In this work, a non-lethal strain of HSV-2 which was isolated clinically was used to infect female mice intravaginally. Body weight, vulval lesions, histological examination of vaginal tissue, and viral load were monitored and used as indices for evaluating antiviral drugs against HSV-2 infection. The results indicated that mice infected with HSV-2 exhibited significant reduction in body weight, serious vulval lesions, massive lymphocyte invasion of vaginal tissue, and approximately 104 copies/µl of HSV-2 were found in vaginal and uterine tissues. Aciclovir (ACV) treatment inhibited loss in body weight, genital pathology and virus replication (reduced to 10°·³ copies/µl) effectively. The study provides a simple, reproducible and feasible animal model for anti-HSV-2 drugs evaluation and HSV-2 vaccine research.