A latent profile analysis of doctors' joy in work at public hospitals.

Introduction: When doctors' work stress increases, their joy in work decreases, severely affecting the quality of care and threatening patient safety. Analysis of the latent categories of joy in work of doctors in public hospitals and the differences in the characteristics of each category can help uncover hidden messages that enhance doctors' joy in work. Methods: Questionnaires were administered to 426 doctors working in public hospitals using the general information questionnaire and the public hospital doctor's joy in work evaluation scale. Upon identifying their potential categories using latent profile analysis, chi-square test, and multinomial logistic regression were performed to analyze the differences in the characteristics of each category. Results: The 426 public hospital doctors could be divided into three potential categories: "low joy in work" (11.27%), "medium joy in work" (59.86%), and "high joy in work" (28.87%). Most of the doctors did not have much joy in work, with 71.13% of them having "low to medium joy in work." Doctors who work in secondary or tertiary hospitals, have a personnel agency or contract, and are older than 45 years are more likely to belong to the "low joy in work" category. Some of the protective factors are having an average monthly income (RMB) of 10,001-15,000 yuan and having a fair or good self-rated health status. Conclusion: There are obvious classification characteristics of doctors' level of joy in work. Hospital managers can take commensurate actions to improve their joy in work, thereby improving patient safety and the quality of medical services.

Genome-wide QTL and eQTL mapping reveal genes associated with growth rate trait of the Pacific white shrimp (Litopenaeus vannamei).

BACKGROUND: Growth rate is a crucial economic trait for farmed animals, but the genetic regulation of this trait is largely unknown in non-model organisms such as shrimp. RESULTS: In this study, we performed genome-wide phenotypic quantitative trait loci (QTL) and expression quantitative trait loci (eQTL) mapping analyses to identify genes affecting the growth rate of Pacific white shrimp (Litopenaeus vannamei), which is the most commercially-farmed crustacean worldwide. We used RNA-sequencing of 268 individuals in a mapping population, and subsequently validated our findings through gene silencing and shrimp growth experiments. We constructed a high-density genetic linkage map comprising 5533 markers spanning 44 linkage groups, with a total distance of 6205.75 cM and an average marker interval of 1.12 cM. Our analyses identified 11 QTLs significantly correlated with growth rate, and 117,525 eQTLs. By integrating QTL and eQTL data, we identified a gene (metalloreductase STEAP4) highly associated with shrimp growth rate. RNA interference (RNAi) analysis and growth experiments confirmed that STEAP4 was significantly correlated with growth rate in L. vannamei. CONCLUSIONS: Our results indicate that the comprehensive analysis of QTL and eQTL can effectively identify genes involved in complex animal traits. This is important for marker-assisted selection (MAS) of animals. Our work contributes to the development of shrimp breeding and available genetic resources.

Genome-wide analysis of ATP-binding cassette (ABC) transporter in Penaeus vannamei and identification of two ABC genes involved in immune defense against Vibrio parahaemolyticus by affecting NF-κB signaling pathway.

The ATP-binding cassette (ABC) transporters have crucial roles in various biological processes such as growth, development and immune defense in eukaryotes. However, the roles of ABC transporters in the immune system of crustaceans remain elusive. In this study, 38 ABC genes were systematically identified and characterized in Penaeus vannamei. Bioinformation analysis revealed that PvABC genes were categorized into ABC A-H eight subfamilies with 17 full-transporters, 11 half transporters and 10 soluble proteins, and multiple immunity-related cis-elements were found in gene promoter regions. Expression analysis showed that most PvABC genes were widely and highly expressed in immune-related tissues and responded to the stimulation of Vibrio parahaemolyticus. To investigate whether PvABC genes mediated innate immunity, PvABCC5, PvABCF1 and PvABCB4 were selected for dsRNA interference experiment. Knockdown of PvABCF1 and PvABCC5 not PvABCB4 increased the cumulative mortality of P. vannamei and bacterial loads in hepatopancreas after infection with V. parahaemolyticus. Further analysis showed that the PvABCF1 and PvABCC5 knockdown decreased expression levels of NF-κB pathway genes and antimicrobial peptides (AMPs). Collectively, these findings indicated that PvABCF1 and PvABCC5 might restrict V. parahaemolyticus challenge by positively regulating NF-κB pathway and then promoting the expression of AMPs, which would contribute to overall understand the function of ABC genes in innate immunity of invertebrates.

Cloning, Identification, and Functional Analysis of the Foxl2 Gene in Procambarus clarkii.

Procambarus clarkii is the most widely distributed freshwater shrimp in China, with important economic value and great potential for development. The forkheadboxL2 (Foxl2) gene has been found to be involved in the reproductive development of many crustaceans. To understand the role of the Foxl2 gene in the gonad development of P. clarkii, we designed CDS-specific primers for the P. clarkii Foxl2 (PcFoxl2) gene and cloned its CDS sequence using RT-PCR. The nucleotide and protein sequence information was then analyzed through bioinformatics analysis. The expression and subcellular localization of PcFoxl2 in various tissues were detected using qRT-PCR and in situ hybridization. The effects of PcFoxl2 knockdown on gonad development were investigated using RNA interference. The results showed that the CDS length of the PcFoxl2 gene was 1614 bp and encoded 537 amino acids. Protein sequence comparison and phylogenetic analysis showed that PcFoxl2 was the closest relative to Crayfish. qRT-PCR analysis indicated that the expression level of PcFoxl2 in the testis was significantly higher (>40 fold) than that in the ovary (p < 0.01). The in situ hybridization results showed that PcFoxl2 was expressed in both the cytoplasm and the nucleus of egg cells, and that the expression was strongest in egg cells at the early stage of yolk synthesis, while weak in the secondary oocytes. The positive signal was strongest in the spermatocyte nucleolus, while only a trace signal was observed in the cytoplasm. After interfering with the PcFoxl2 gene using dsRNA, the expression of PcFoxl2 in the RNA interference group was significantly lower than that in the control group, and this interference effect lasted for one week. Moreover, the gonad index of the experimental group was significantly lower than that of the control group (p < 0.05) after 10 days of P. clarkii cultivation following PcFoxl2 knockdown. The expression levels of the nanos and S3a genes, which are related to gonad development, decreased significantly after PcFoxl2 gene interference. The results suggest that the Foxl2 gene is involved in the growth and development of gonads, particularly in the development of testis, and is related to the early development of oocytes. This study provides a theoretical basis for the artificial breeding of P. clarkii.

Interkingdom ecological networks between plants and fungi drive soil multifunctionality across arid inland river basin.

Despite the known collective contribution of above- (plants) and below-ground (soil fungi) biodiversity on multiple soil functions, how the associations among plant and fungal communities regulate soil multifunctionality (SMF) differentially remains unknown. Here, plant communities were investigated at 81 plots across a typical arid inland river basin, within which associated soil fungal communities and seven soil functions (nutrients storage and biological activity) were measured in surface (0-15 cm) and subsurface soil (15-30 cm). We evaluated the relative importance of species richness and biotic associations (reflected by network complexity) on SMF. Our results demonstrated that plant species richness and plant-fungus network complexity promoted SMF in surface and subsurface soil. SMF in two soil layers was mainly determined by plant-fungus network complexity, mean groundwater depth and soil variables, among which plant-fungus network complexity played a crucial role. Plant-fungus network complexity had stronger effects on SMF in surface soil than in subsurface soil. We present evidence that plant-fungus network complexity surpassed plant-fungal species richness in determining SMF in surface and subsurface soil. Moreover, plant-fungal species richness could not directly affect SMF. Greater plant-fungal species richness indirectly promoted SMF since they ensured greater plant-fungal associations. Collectively, we concluded that interkingdom networks between plants and fungi drive SMF even in different soil layers. Our findings enhanced our knowledge of the underlying mechanisms that above- and below-ground associations promote SMF in arid inland river basins. Future study should place more emphasis on the associations among plant and microbial communities in protecting soil functions under global changes.

Constraints on the maintenance of wild Taxus cuspidata populations based on regression conservation.

To clarify the key factors constraining the maintenance of wild Taxus cuspidata populations and to develop conservation strategies and technical links for current populations, we investigated the renewal status and distribution patterns of wild T. cuspidata populations in the main distribution areas of China. We analyzed the effects of stand factors and human disturbance on population renewal and maintenance. The results showed that the overall regeneration of wild T. cuspidata populations was poor. The basal diameter and height class structure of renewed individuals showed an unhealthy state. 19% of the area was well regenerated. There were three types of regeneration, including poor regeneration with few adult trees, poor regeneration with many adult trees, and good regeneration with few adult trees. The communities in which T. cuspidata was found could be classified into Abies nephrolepis + Tilia amurensis forest, spinney forest, and Picea jezoensis var. microsperma + A. nephrolepis forest. The renewal number of A. nephrolepis + T. amurensis forest was significantly higher than that of spinney forest. Increased stand density and moderate human disturbance contributed to the regeneration of T. cuspidata. The regenerating T. cuspidata seedlings increased significantly when stand density increased from low to medium. The number of regenerating populations in moderately disturbed habitats was significantly higher than those in lightly disturbed habitats. Human disturbance and habitat were currently critical constraints to maintaining and regenerating wild T. cuspidata populations. The conservation of T. cuspidata should consider current status of population regeneration in each habitat patch to develop corresponding in situ conservation and regression conservation measures and focus on the influence of critical factors such as disturbances and habitat conditions.

Integrated Analysis of DNA Methylome and Transcriptome Reveals Epigenetic Regulation of Cold Tolerance in Litopenaeus vannamei.

DNA methylation is an important epigenetic modification that has been shown to be associated with responses to non-biological stressors. However, there is currently no research on DNA methylation in response to environmental signals in shrimp. In this study, we conducted a comprehensive comparative analysis of DNA methylation profiles and differentially expressed genes between two strains of Litopenaeus vannamei with significantly different cold tolerance through whole genome bisulfite sequencing (WGBS) and transcriptome sequencing. Between Lv-C and Lv-T (constant temperature of 28 °C and low temperatures of 18 °C and 10 °C) under cytosine-guanine (CG) environments, 39,100 differentially methylated regions (DMRs) were identified, corresponding to 9302 DMR-related genes (DMRGs). The DMRs were mainly located in the gene body (exons and introns). Gene Ontology (GO) analysis showed that these DMRGs were significantly enriched in cell parts, catalytic activity, and metabolic processes. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed significant enrichment of these DMRGs in pathways such as proteasome (ko03050), oxidative phosphorylation (ko00190), mTOR signaling pathway (ko04150), fatty acid metabolism (ko01212), and fatty acid degradation (ko00071). The comprehensive results suggested that L. vannamei mainly regulates gene expression in response to low temperatures through hypermethylation or demethylation of some genes involved in thermogenesis, glycolysis, the autophagy pathway, the peroxisome, and drug metabolism pathways. These results provide important clues for studying DNA methylation patterns and identifying cold tolerance genes in shrimp.

The role of phytomelatonin receptor 1-mediated signaling in plant growth and stress response.

Phytomelatonin is a pleiotropic signaling molecule that regulates plant growth, development, and stress response. In plant cells, phytomelatonin is synthesized from tryptophan via several consecutive steps that are catalyzed by tryptophan decarboxylase (TDC), tryptamine 5-hydroxylase (T5H), serotonin N-acyltransferase (SNAT), and N-acetylserotonin methyltransferase (ASMT) and/or caffeic acid-3-O-methyltransferase (COMT). Recently, the identification of the phytomelatonin receptor PMTR1 in Arabidopsis has been considered a turning point in plant research, with the function and signal of phytomelatonin emerging as a receptor-based regulatory strategy. In addition, PMTR1 homologs have been identified in several plant species and have been found to regulate seed germination and seedling growth, stomatal closure, leaf senescence, and several stress responses. In this article, we review the recent evidence in our understanding of the PMTR1-mediated regulatory pathways in phytomelatonin signaling under environmental stimuli. Based on structural comparison of the melatonin receptor 1 (MT1) in human and PMTR1 homologs, we propose that the similarity in the three-dimensional structure of the melatonin receptors probably represents a convergent evolution of melatonin recognition in different species.

Identifying Emissions Reduction Opportunities in International Bilateral Emissions Trading Systems to Achieve China's Energy Sector NDCs.

Exploring more emissions reduction opportunities for China's energy sector and lowering China's decarbonisation costs are essential to fulfilling China's nationally determined contributions (NDCs) and making China's sustainable development more feasible. This study explored emissions reduction opportunities for China's energy sector in international bilateral emissions trading systems (ETSs) using a CGE (computable general equilibrium) model. This study revealed that linking China's ETS to those of regions with lower decarbonisation responsibilities, which tend to be developing regions, could lower China's carbon prices, thus increasing China's domestic energy supply and lowering energy prices (and vice versa). Meanwhile, the volume of emissions from regions linked to China also significantly affected the degree of the change in China's carbon prices. Among these, ETS links to India and Russia could reduce China's carbon price from 7.80 USD/ton under domestic ETS to 2.16 USD/ton and 6.79 USD/ton, allowing the energy sector and energy-intensive sectors to increase greenhouse gas emissions by 1.14% and 7.05%, respectively, without falling short of meeting its NDC targets. In contrast, as a consequence of links with the United States and the European Union, China's carbon price could increase to 5.37 USD/ton and 1.79 USD/ton, respectively, which would limit China's energy and energy-intensive sectors to emitting 5.45% and 2.24% fewer greenhouse gases in order to meet its NDC targets.

The variation on structure and immunomodulatory activity of polysaccharide during the longan pulp fermentation.

In the current study, the effects of fermentation manners on the structure and immunomodulatory activity of polysaccharide in longan wine or vinegar were investigated. Compared to longan polysaccharide (CP1), polysaccharide in longan wine (CP2) or vinegar (CP3 and CP4) had smaller molecular weights, and was consisted of more mannose, arabinose, rhamnose, galactose and less glucose. After purification, the major fraction (P1-P4) was obtained from CP1-CP4, respectively. The structures and immunoregulatory activities of P1-P4 were characterized. Fermentation and purification were favorable to increase the immunoregulatory activities of P2-P4, which were contributed to their different structural features. The structure-activity relationship analysis indicated that molecular weight, mannose, rhamnose, glucuronic acid, glucose and arabinose were significantly associated with the cytokines secretion. Compared with other polysaccharides, P3 displayed better immunomodulatory activity due to its lower molecular weight, lower contents of rhamnose and glucose, and higher levels of mannose and arabinose by activating MAPK and PI3K/Akt signaling pathways.

Trend Analysis of the Mortality Rates of the Top Three Causes of Death Among Chinese Residents from 2003 to 2019.

Objective: To delineate the mortality trends of malignant tumors, heart disease and cerebrovascular disease in China. Methods: The crude mortality from 2003 to 2019 was derived from the China Health Statistical Yearbook, and the mortality rates were analyzed through joinpoint regression supplemented by descriptive statistics and χ2 tests. Results: The fitting model of age-standardized mortality due to malignant tumors showed three joinpoints. The APCs from 2003 to 2005, 2005-2008, 2008-2012 and 2012-2019 were -11.00%, 9.63%, -4.67% and -1.40%, respectively, and the AAPC was -1.54%. The mortality rate of cerebrovascular disease consistently decreased (APC = AAPC = -0.98%). In the subgroup analyses, significant differences were observed between sexes and regions. The mortality rate of heart disease among rural females exhibited an upward trend (APC = AAPC = 2.33%). Older adults aged over 75 years had the highest mortality rates and the most drastic change. Conclusion: The three diseases had variable change trends. The government should focus more on policies that promote the equalization of basic public health services. Continuous education on heart disease, which includes not only beneficial behaviors but also knowledge of first aid, should be strengthened for rural females.

Influence of Provenance and the Sedimentary Environment on the Formation of High-Quality Source Rocks in the Paleocene Lishui Sag, China.

Organic matter (OM) is the material basis for hydrocarbon generation. Its type and abundance determine the hydrocarbon generation ability of source rocks, which is closely related to the provenance and sedimentary environment of source rocks. The tectonic backgrounds of the eastern and western subsags (ESS and WSS) of the Lishui Sag in the East China Sea Shelf Basin are significantly different and their influence on the OM in the source rocks is worthy of attention. This paper comprehensively analyzes the provenance and environmental characteristics and their influence on the features of the OM of Paleocene source rocks in the ESS and WSS. The study finds that the source rocks in the ESS have multiple sources. During the deposition period, as the salinity and paleoproductivity of the water column increased, the proportion of OM in the autochthonous components of the water column continued to increase, but the overall water column was in an oxidizing environment, resulting in a generally low abundance of OM. The provenance of the source rocks in the WSS was relatively simple and terrestrial. Also, the sedimentary environment had little effect on the type of OM. However, the whole water column of the WSS was in an anoxic environment, so the OM was better preserved, resulting in a higher abundance. Due to the influence of provenance and the sedimentary environment in different areas of the sag, the characteristics of OM in the source rocks are different, so relevant exploration strategies need to be adopted in actual exploration.

Single-Cell Ribonucleic Acid Sequencing Clarifies Cold Tolerance Mechanisms in the Pacific White Shrimp (Litopenaeus Vannamei).

To characterize the cold tolerance mechanism of the Pacific white shrimp (Litopenaeus vannamei), we performed single-cell RNA sequencing (scRNA-seq) of ∼5185 hepatopancreas cells from cold-tolerant (Lv-T) and common (Lv-C) L. vannamei at preferred and low temperatures (28°C and 10°C, respectively). The cells fell into 10 clusters and 4 cell types: embryonic, resorptive, blister-like, and fibrillar. We identified differentially expressed genes between Lv-T and Lv-C, which were mainly associated with the terms "immune system," "cytoskeleton," "antioxidant system," "digestive enzyme," and "detoxification," as well as the pathways "metabolic pathways of oxidative phosphorylation," "metabolism of xenobiotics by cytochrome P450," "chemical carcinogenesis," "drug metabolism-cytochrome P450," and "fatty acid metabolism." Reconstruction of fibrillar cell trajectories showed that, under low temperature stress, hepatopancreas cells had two distinct fates, cell fate 1 and cell fate 2. Cell fate 1 was mainly involved in signal transduction and sensory organ development. Cell fate 2 was mainly involved in metabolic processes. This study preliminarily clarifies the molecular mechanisms underlying cold tolerance in L. vannamei, which will be useful for the breeding of shrimp with greater cold tolerance.

A Scale to Measure the Joy in Work of Doctors: Development, Validity, and Reliability.

Background: The aim of this study is to develop a scale and evaluate its' validity and reliability to measure the joy in work of doctors. Methods: Based on literature review and panel discussion, the scale framework and item pool were determined. Next, the items were modified by two rounds of expert consultation. Then the pre-investigation was applied and the formal version of scale was formed. Last, the reliability and validity of the scale were tested with 426 physicians. Results: The scale was composed of four dimensions: work autonomy needs, competency identification needs, competency perception needs and work relationship needs. Each dimension had 7 items, and both reliability and validity were acceptable. The Cronbach α coefficient and half-reliability coefficient of the whole scale were 0.954 (>0.9) and 0.974 (>0.9). The Spearman correlations of item-total score ranged from 0.556 to 0.749, indicating a good-item total score correlation. The χ 2/ df, RMSEA, RMR, GFI, CFI, and TLI, CFA of the maximum likelihood method supported a good fit with the model. Conclusions: Based on the self-determination theory, this study develops a scale to measure the joy in work of doctors. It has good validation and reliability, which is useful for doctors and medical institutions to take steps to improve happiness.

Identification of a quantitative trait loci (QTL) associated with ammonia tolerance in the Pacific white shrimp (Litopenaeus vannamei).

BACKGROUND: Ammonia is one of the most common toxicological environment factors affecting shrimp health. Although ammonia tolerance in shrimp is closely related to successful industrial production, few genetic studies of this trait are available. RESULTS: In this study, we constructed a high-density genetic map of the Pacific white shrimp (Litopenaeus vannamei) using specific length amplified fragment sequencing (SLAF-seq). The constructed genetic map contained 17,338 polymorphic markers spanning 44 linkage groups, with a total distance of 6360.12 centimorgans (cM) and an average distance of 0.37 cM. Using this genetic map, we identified a quantitative trait locus (QTL) that explained 7.41-8.46% of the phenotypic variance in L. vannamei survival time under acute ammonia stress. We then sequenced the transcriptomes of the most ammonia-tolerant and the most ammonia-sensitive individuals from each of four genetically distinct L. vannamei families. We found that 7546 genes were differentially expressed between the ammonia-tolerant and ammonia-sensitive individuals. Using QTL analysis and the transcriptomes, we identified one candidate gene (annotated as an ATP synthase g subunit) associated with ammonia tolerance. CONCLUSIONS: In this study, we constructed a high-density genetic map of L. vannamei and identified a QTL for ammonia tolerance. By combining QTL and transcriptome analyses, we identified a candidate gene associated with ammonia tolerance. Our work provides the basis for future genetic studies focused on molecular marker-assisted selective breeding.

Construction of a High-Density Genetic Map and Identification of Quantitative Trait Loci for Nitrite Tolerance in the Pacific White Shrimp (Litopenaeus vannamei).

Nitrite is a major environmental toxin in aquaculture systems that disrupts multiple physiological functions in aquatic animals. Although nitrite tolerance in shrimp is closely related to successful industrial production, few genetic studies of this trait are available. In this study, we constructed a high-density genetic map of Litopenaeus vannamei with 17,242 single nucleotide polymorphism markers spanning 6,828.06 centimorgans (cM), with an average distance of 0.4 cM between adjacent markers on 44 linkage groups (LGs). Using this genetic map, we identified two markers associated with nitrite tolerance. We then sequenced the transcriptomes of the most nitrite-tolerant and nitrite-sensitive individuals from each of four genetically distinct L. vannamei families (LV-I-4). We found 2,002, 1,983, 1,954, and 1,867 differentially expressed genes in families LV-1, LV-2, LV-3, and LV-4, respectively. By integrating QTL and transcriptomics analyses, we identified a candidate gene associated with nitrite tolerance. This gene was annotated as solute carrier family 26 member 6 (SLC26A6). RNA interference (RNAi) analysis demonstrated that SLC26A6 was critical for nitrite tolerance in L. vannamei. The present study increases our understanding of the molecular mechanisms underlying nitrite tolerance in shrimp and provides a basis for molecular-marker-assisted shrimp breeding.

Penaeus stylirostris densovirus proteins CP and NS1 interact with peritrophin of Litopenaeus vannamei.

The Penaeus stylirostris densovirus (PstDNV) is a major virus of shrimps that severely harms the shrimp farming industry. Peritrophin is a peritrophic membrane protein with chitin binding activity. To examine the roles of peritrophin in viral infection, we used yeast two-hybrid to analyze the interaction between the Pacific white shrimp (Litopenaeus vannamei) peritrophin and PstDNV proteins (CP, NS1 and NS2). The yeast two-hybrid results showed that NS1 and peritrophin had an interaction, CP and peritrophin had an interaction as well, and NS2 had no interaction with peritrophin. We validated the interactions with GST pull-down assays. We then conducted RNA interference and qRT-PCR. The results showed that when pre-injection of dsRNA-peritrophin, the quantity of PstDNV in the shrimps injected with viruses was significantly lower than in the control group (P < 0.01), indicating the viral infection was decreased when the peritrophin gene expression was inhibited. The results indicated that peritrophin of L. vannamei participated in the PstDNV infection.

Integrated analysis of physiological, transcriptomic and metabolomic responses and tolerance mechanism of nitrite exposure in Litopenaeus vannamei.

Nitrite accumulation in aquatic environments is a potential risk factor that disrupts multiple physiological functions in aquatic animals. In this study, the physiology, transcriptome and metabolome of the control group (LV-C), nitrite-tolerance group (LV-NT) and nitrite-sensitive group (LV-NS) were investigated to identify the stress responses and mechanisms underlying the nitrite tolerance of Litopenaeus vannamei. After LV-NT and LV-NS were subjected to nitrite stress, the hemocyanin contents were significantly decreased, and hepatopancreas showed severe histological damage compared with LV-C. Likewise, the antioxidant enzymes were also significantly changed after nitrite exposure. The transcriptome data revealed differentially expressed genes associated with immune system, cytoskeleton remodeling and apoptosis in LV-NT and LV-NS. The combination of transcriptomic and metabolomic analysis revealed nitrite exposure disturbed metabolism processes in L. vannamei, including amino acid metabolism, nucleotide metabolism and lipid metabolism. The multiple comparative analysis implicated that higher nitrite tolerance of LV-NT than LV-NS may be attributed to enhanced hypoxia inducible factor-1α expression to regulate energy supply and gaseous exchange. Moreover, LV-NT showed higher antioxidative ability, detoxification gene expression and enhanced fatty acids contents after nitrite exposure in relative to LV-NS. Collectively, all these results will greatly provide new insights into the molecular mechanisms underlying the stress responses and tolerance of nitrite exposure in L. vannamei.

Immune Responses to Gram-Negative Bacteria in Hemolymph of the Chinese Horseshoe Crab, Tachypleus tridentatus.

Chinese horseshoe crab, Tachypleus tridentatus, is an ancient marine arthropod with a long evolutionary history. As a kind of living fossil species, the pathogen defenses of horseshoe crabs entirely depend on the innate immune system. Although, there are abundant immune molecules found in the horseshoe crab hemolymph, the biological mechanisms underlying their abilities of distinguishing and defending against invading microbes are still unclear. In this study, we used high-throughput sequencing at mRNA and protein levels and bioinformatics analysis methods to systematically analyze the innate immune response to Gram-negative bacteria in hemolymph of Chinese horseshoe crab. These results showed that many genes in the complement and coagulation cascades, Toll, NF-κB, C-type lectin receptor, JAK-STAT, and MAPK signaling pathways, and antimicrobial substances were activated at 12 and 24 h post-infection, suggesting that Gram-negative bacteria could activate the hemolymph coagulation cascade and antibacterial substances release via the above pathways. In addition, we conjectured that Toll and NF-κB signaling pathway were most likely to participate in the immune response to Gram-negative bacteria in hemolymph of horseshoe crab through an integral signal cascade. These findings will provide a useful reference for exploring the ancient original innate immune mechanism.