Comparative evaluation of soundscapes in human activities spatial contexts of pedestrian spaces adjacent to arterial roads.

Pedestrian spaces adjacent to arterial roads are characterized by the dominance of traffic noise alongside various human activities. Research on the impact of traffic noise on the soundscape evaluation of pedestrian spaces has not considered human activities spatial contexts. To address this research gap, the present study constructed auditory environments for pedestrian spaces in the contexts of commuting, residential, and commercial activities. A total of seven auditory environments were subjected to laboratory auditory evaluations, including perceived dominance of sound source, acoustic comfort, and perceived affective quality of the soundscape. The results indicated that in pedestrian spaces with constant traffic noise, the presence of significant human activity sounds led to a decreased perceived dominance of traffic noise and an increased acoustic comfort, despite the higher acoustic energy. Thus, pedestrian spaces with a variety of human activity received better soundscape evaluations. The elements that reflected the human activities spatial contexts, including the types and intensity of human activities, played a crucial role in soundscape evaluations. Better acoustic comfort was reported in pedestrian spaces characterized by low-intensity residential activities and high-intensity commercial activities. Additionally, pedestrian spaces with more intense activities offered an actively engaging soundscape. The findings can provide reference for a more accurate evaluation of the soundscape in pedestrian spaces and guide the soundscape design of pedestrian environments.

25-hydroxyvitamin D and Endometriosis: A bidirectional Mendelian randomization study.

Numerous studies have demonstrated a correlation between serum 25-hydroxyvitamin D (25OHD) and endometriosis. However, the precise nature of this association remains elusive. The causal connection between 25OHD and endometriosis remains uncertain, as it is yet to be determined whether one directly influences the other. The objective of our research was to investigate the cause-and-effect connection between 25OHD and endometriosis. The study employed Mendelian randomization (MR) in a bidirectional two-sample investigation to examine the causal relationship between 25OHD and endometriosis. The analysis utilized the most recent publicly accessible statistics from the genome-wide association study (GWAS) encompassing 25OHD, endometriosis, and its five subtypes. The primary analytical approach employed was Inverse-Variance Weighting (IVW), accompanied by supplementary analysis methods including weighted median, MR-Egger, simple mode, and weighted mode. Furthermore, sensitivity analyses were conducted to assess the potential influence of heterogeneity and pleiotropy on the MR outcomes. MR primary analysis showed no significant causal effect of 25OHD on endometriosis (OR = 0.892, 95%CI = 0.745 ~ 1.068, P = 0.213). Similarly, there was no evidence to support a causal relationship of endometriosis on 25OHD (IVW Beta = 0.005, 95%CI = 0.993 ~ 1.018, P = 0.406). However, when conducting MR analysis on different subtypes of endometriosis and 25OHD, we found a positive correlation between endometriosis of ovary and 25OHD level (IVW Beta = 0.012, 95%CI = 1.002 ~ 1.022, P = 0.024). This study indicates that there is no causal relationship between serum 25OHD and endometriosis. However, it is important to note that serum 25OHD levels will increase in patients with endometriosis of the ovary. Further observational studies and clinical trials are indispensable.

Comparative analysis of whole-transcriptome RNA expression of lung tissue of Chinese soft-shell turtle infected by Trionyx sinensis Hemorrhagic Syndrome Virus.

Trionyx sinensis Hemorrhagic Syndrome Virus (TSHSV), the first aquatic arterivirus identified in China, causes severe mortality to T. sinensis. In this study, we sought to determine the functions of T. sinensis mRNAs and non-coding RNAs (ncRNAs) that were differentially expressed (DE) over different periods of TSHSV infection of T. sinensis lung. We used RT-qPCR to validate the sequencing results of select RNAs, confirming their reliable and referable nature. Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were used to predict multiple biological functions and signaling pathways in various comparison groups (1-day versus mock, 3-day versus 1-day, and 5-day versus 3-day). Multiple types of differentially expressed RNA, including mRNA, lncRNA, circRNA, and miRNA, were associated with cardiac dysfunction, coagulation abnormalities, and arachidonic acid metabolism at day 1. Pre-inflammatory cytokines and inflammatory factors such as PLA2G4A, cPLA2, γ-GGT1, TNFRSF14, TCP11L2, PTER CYP2J2 and LTC4S, were noticeably regulated at the same time. On day 3, multiple GO terms and KEGG pathways were implicated, including those related to virus defense, apoptosis, pyroptosis, and inflammatory response. Notably, key genes such as RSAD2, TRIM39, STAT4, CASP1, CASP14, MYD88, CXCL3, CARD11, ZBP1, and ROBO4 exhibited significant regulation. The lncRNAs and circRNAs that targeted the genes involved in viral recognition (TLR5), apoptosis (CARD11), pyroptosis (ZBP1), inflammatory processes (NEK7, RASGRP4, and SELE) and angiogenesis (ROBO4) exhibited significant regulation. Significantly regulated miRNAs were primarily linked to genes involved in apoptosis (Let-7f-3p, miR-1260a, miR-455-3p), and inflammation (miR-146a, miR-125a, miR-17a, miR-301b, and miR-30a-3p). The findings could advance our understanding of the host immunological response to TSHSV and offer new ideas for developing effective strategies to prevent infection of T. sinensis.

Diversity of MrTolls and their regulation of antimicrobial peptides expression during Enterobacter cloacae infection in Macrobrachium rosenbergii.

Toll/Toll-like receptor (TLR) is an important pattern recognition receptor that plays an important role in the immunity of animals. Six Toll genes were identified in Macrobrachium rosenbergii, namely, MrToll, MrToll1, MrToll2, MrToll3, MrToll4, and MrToll5. SMART analysis showed that all six Tolls have a transmembrane domain, a TIR domain, and different number of LRR domains. The phylogenetic tree showed that six Tolls were located in six different branches. Among these six Tolls, only MrToll4 contains the QHR motif, which is similar to insect Toll9. MrToll4 belongs to V-type/scc Toll with only one LRRCT domain. MrToll1 and MrToll5 are classical P-type/mcc Toll with two LRRCT domains and an LRRNT. MrTolls were distributed in the hemocytes, heart, hepatopancreas, gills, stomach, and intestine. During the infection of Enterobacter cloacae, the expression level of MrToll and MrToll1-4 was upregulated in the intestine of M. rosenbergii. RNA interference experiments showed that the expression of most antimicrobial peptide (AMP) genes was negatively regulated by MrTolls during E. cloacae infection. On the contrary, crustin (Cru) 3 and Cru4 were inhibited after the knockdown of MrToll, and Cru1 and Cru4 were significantly downregulated with the knockdown of MrToll4 during E. cloacae challenge. These results suggest that MrTolls may be involved in the regulation of AMP expression in the intestine during E. cloacae infection.

FOXO is involved in antimicrobial peptides expression during WSSV infection in Exopalaemon carinicauda.

The forkhead box transcription factor O family protein (FOXO) acts as a transcription factor that regulates biological processes regarding DNA repair, immunity, cell cycle regulation, and other biological processes. In this study, EcFOXO was identified from the ridgetail white prawn, Exopalaemon carinicauda. EcFOXO protein contains multiple low-complexity regions and a forkhead (FH) domain. Phylogenetic tree showed that EcFOXO is clustered with crustacean FOXOs. The amino acid sequences of its FH domain are highly similar to the FH domain of FOXOs from other crustaceans. The expression of EcFOXO is altered after white spot syndrome virus (WSSV) stimulation in hepatopancreas and gills. The relationship between EcFOXO and EcRelish was explored by RNA interference (RNAi). Results showed that EcFOXO and EcRelish could positively regulate each other's expression. The expression levels of various antimicrobial peptides (AMPs) significantly reduced after interfering with EcFOXO or EcRelish. These results suggest a positive regulatory loop between EcFOXO and EcRelish, which participates in the innate immunity of ridgetail white prawn by regulating the expression of AMPs during WSSV infection. This study enriches the knowledge about the regulatory mechanism of FOXO in the innate immunity of crustaceans.

Lectin diversity and their positive roles in WSSV replication through regulation of calreticulin expression and inhibiting ALFs expression.

In biological evolution, gene duplication (GD) generates new genes to facilitate new functions. C-type lectins (CTLs) in crayfish have been extended by GD to expand their family members. In this study, four CTL genes generated by GD were identified from Procambarus clarkii (PcLec1-4). Among these four genes, PcLec1 can also generate new isoforms with different numbers of tandem repeats through DNA slip mispairing. PcLec1-4 was widely expressed in multiple tissues. The expression levels of PcLec1-4 were upregulated in the intestine of P. clarkii upon white spot syndrome virus (WSSV) challenge at multiple time points. Further analysis indicated that GATA transcription factor regulated PcLec1-4 expression. RNA interference and recombinant PcLec1-4 protein injection experiments suggested that PcLec1-4 promoted the expression of calreticulin (PcCRT) and negatively regulated the expression of antimicrobial peptides, thereby promoting WSSV replication. This study contributes to the understanding of the function of CTLs produced by GD during WSSV invasion in crustaceans.

Three STAT isoforms formed by selective splicing are involved in the regulation of anti-lipopolysaccharide factor expression in Macrobrachium nipponense during WSSV infection.

White spot syndrome virus (WSSV), a double-stranded DNA virus, is harmful in aquaculture. The signal transducer and activator of transcription (STAT) has been shown to play a role during host infection with the virus, but the exact mechanism by which it acts is unclear. In this study, three STAT isoforms (MnSTAT1, MnSTAT2, and MnSTAT3) were identified in Macrobrachium nipponense. The full-length sequence of MnSTAT1 was 3336 bp, with 2259 bp open reading frame (ORF), encoding a 852 amino acids protein. The full-length sequence of MnSTAT2 was 2538 bp, and the ORF was 2391 bp, encoding 796 amino acids. The full-length sequence of MnSTAT3 sequence was 2618 bp, and the ORF was 2340 bp, encoding 779 amino acids. MnSTAT1-3 is produced by alternative last exon. MnSTAT1-3 all contain a STAT_int, a STAT_alpha, a STAT_bind, and a SH2 structure. MnSTAT1-3 are widely expressed in various tissues tested. The expression levels of MnSTAT1-3 in the intestine of M. nipponense were upregulated at multiple time points following WSSV stimulation. The expression of seven anti-lipopolysaccharide factors (ALFs) was significantly reduced with the knockdown of MnSTATs during WSSV infection. Results showed that MnSTATs regulated the expression of intestinal ALFs and was involved in the innate immunity against WSSV of M. nipponense.

Duox mediated ROS production inhibited WSSV replication in Eriocheir sinensis under short-term nitrite stress.

Nitrite stress and white spot syndrome virus (WSSV) infection are major problems threatening the sustainable and healthy development of Eriocheir sinensis. Some studies have found that nitrite stress can lead to the production of reactive oxygen species (ROS), whereas synthetic ROS plays a vital role in the signaling pathway. However, whether nitrite stress influences the infection of crabs by WSSV remains unclear. NADPH oxidases, including NOX1-5 and Duox1-2, are important for ROS production. In the present study, a novel Duox gene (designated as EsDuox) was identified from E. sinensis. The studies found that nitrite stress could increase the expression of EsDuox during WSSV infection and decrease the transcription of the WSSV envelope protein VP28. Moreover, nitrite stress could increase the production of ROS, and the synthesis of ROS relied on EsDuox. These results indicated a potential "nitrite stress-Duox activation-ROS production" pathway that plays a negative role in WSSV infection in E. sinensis. Further studies found that nitrite stress and EsDuox could promote the expression of EsDorsal transcriptional factor and antimicrobial peptides (AMPs) during WSSV infection. Moreover, the synthesis of AMPs was positively regulated by EsDorsal in the process of WSSV infection under nitrite stress. Furthermore, EsDorsal played an inhibitory role in the replication of WSSV under nitrite stress. Our study reveals a new pathway for "nitrite stress-Duox activation-ROS production-Dorsal activation-AMP synthesis" that is involved in the defense against WSSV infection in E. sinensis during short-term nitrite stress.

A positive feedback loop between two C-type lectins originated from gene duplication and relish promotes the expression of antimicrobial peptides in Procambarus clarkii.

Gene duplication (GD) leads to the expansion of gene families that contributes organisms adapting to stress or environment and dealing with the infection of various pathogens. C-type lectins (CTLs) in crustaceans undergo gene expansion and participate in various immune responses. However, the functions of different CTL produced by GD are not fully characterized. In the present study, two CTL genes (designated as PcLec-EPS and PcLec-QPS, respectively) were identified from Procambarus clarkii. PcLec-EPS and PcLec-QPS originate from GD and the main difference between them is exon 3. PcLec-EPS and PcLec-QPS respectively contains EPS and QPS motif in their carbohydrate recognition domain. The mRNA levels of PcLec-EPS and PcLec-QPS in hemocytes, gills, intestine and lymph underwent time-dependent enhancement after D-Mannose and D-Galactose challenge. Recombinant PcLec-EPS and PcLec-QPS could bind to carbohydrates and microbes, and agglutinate bacteria. The results of experiments on recombinant protein injection and RNA interference indicate that PcLec-EPS and PcLec-QPS can respectively strong recognize and bind D-Mannose and D-Galactose, activate the Relish transcriptional factor, and further upregulate the expression of different antimicrobial peptides (AMPs). In addition, these two CTLs and Relish could positively regulate the expression of each other, suggesting that there is a positive feedback loop between two CTLs and Relish that regulates the expression of AMPs. It may contribute to the expansion of the immune response for host quickly and efficiently eliminating pathogenic microorganisms. This study provides new knowledge for clear understanding the significance and function of different CTL generated by GD in immune defenses in crustacean.

Identification of three novel Spätzle genes in Eriocheir sinensis and their roles during white spot syndrome virus infection.

Proteins of Spätzle family play an essential role in innate immunity in invertebrates by activating the Toll pathway to induce the expression of antimicrobial peptides. However, little is known about the function of Spätzle in in the immune response of the Chinese mitten crab. In the present study, three novel Spätzle genes (named as EsSpz1, EsSpz2, and EsSpz3) were identified from Eriocheir sinensis. The genome structure of EsSpz1 contains two exons and an intron. Three Spätzle proteins all contain a Pfam Spaetzle domain. In the evolution, EsSpz1-3 cluster with other Spätzle proteins from crustaceans. EsSpz1-3 were widely distributed in multiple immune tissues. The expression levels of EsSpz1-3 in the intestine were remarkably upregulated after white spot syndrome virus (WSSV) challenge. The knockdown of EsSpz1-3 remarkably decreased the expressions of crustins and anti-lipopolysaccharide factors during WSSV infection. Moreover, EsSpz1-3 silencing remarkably increased the expression of WSSV envelope protein VP28. These findings suggest that new-found EsSpz1-3 in E. sinensis could promote the synthesis of antimicrobial peptides and inhibit the expression of VP28 during WSSV infection. Our study indicates that EsSpz1-3 in E. sinensis may participate in the innate immune defenses against WSSV by inducing the expression of antimicrobial peptides. This study provides new knowledge for the function of Spätzle in the antiviral immune defense in crustacean.

Characterization of four spliced isoforms of a transmembrane C-type lectin from Procambarus clarkii and their function in facilitating WSSV infection.

C-type lectin (CTL) is an important pattern recognition receptor that play vital functions in the innate immunity. Many soluble CTLs in crustacean participate in the inhibition or promotion of white spot syndrome virus (WSSV) infection. However, whether transmembrane CTLs participate in WSSV infection in crustacean remains unknown. In the present study, four spliced isoforms of a transmembrane CTL (designated as PcTlec) from Procambarus clarkii were identified for the first time. The genome structure of PcTlec contains eight exons, six known introns, and one unknown intron. PcTlec-isoform1 is produced by intron retention, whereas PcTlec-isoform3 and PcTlec-isoform4 are produced by exon skipping. All of them contain the transmembrane domain and characteristic carbohydrate recognition domain (CRD). Four PcTlec isoforms were mainly expressed in the hepatopancreas, stomach, and intestine. After WSSV challenge, the expression levels of PcTlec-isoform1-4 in the intestine were upregulated. The knockdown of the region shared by four PcTlec isoforms evidently decreased the expression of WSSV envelope protein VP28 and the copies of viral particles. A recombinant protein (rPcTlec-CRD) containing the CRD that was shared by four PcTlec isoforms was acquired by procaryotic expression system. The injection of purified rPcTlec-CRD protein evidently increased the VP28 expression and WSSV copies during viral infection. Moreover, rPcTlec-CRD could directly bind to WSSV and interact with VP28 protein. These findings indicate that new-found transmembrane CTL isoforms in P. clarkii may act as viral receptors that facilitate WSSV infection. This study contributes to the recognition and understanding of the functions of transmembrane CTLs in crustacean in the infection of host by WSSV.

Characterization and functional analysis of a clip domain serine protease (MncSP) and its alternative transcript (MncSP-isoform) from Macrobrachium nipponense.

Clip domain serine protease (cSPs) play an important role in the innate immune defense of crustaceans. In this study, a clip domain serine protease (MncSP) and its alternative transcript (MncSP-isoform) were identified from Macrobrachium nipponense. The full-length cDNA sequences of MncSP and MncSP-isoform were 2447 and 2351 bp with open reading frames comprising 1497 and 1401 bp nucleotides and encoding 498 and 466 amino acids, respectively. The genome of MncSP had 10 exons and 9 introns. MncSP contained all 10 exons, whereas MncSP-isoform lacked the second exon. MncSP and MncSP-isoform contained a signal peptide, a clip domain, and a Tryp_SPc domain. Phylogenetic tree analysis showed that MncSP and MncSP-isoform clustered with cSPs from Palaemonidae. MncSP and MncSP-isoform were widely distributed in hemocytes, heart, hepatopancreas, gills, stomach, and intestine. The expression profiles of MncSP and MncSP-isoform in the hemocytes of M. nipponense changed after simulation by Vibrio parahaemolyticus or Staphylococcus aureus. The RNAi of MncSP could inhibit the expression of antimicrobial peptides (AMPs), including crustins and anti-lipopolysaccharide factors. Phenoloxidase activity was also down-regulated in MncSP-silenced prawns. This study indicated that MncSP participated in the synthesis of AMPs and the activation of prophenoloxidase.

The prevalence and risk factors for anxiety in frontline nurses under COVID-19 pandemic based on a large cross-sectional study using the propensity score-matched method

ABSTRACT Introduction: We determined the prevalence of anxiety and the associated risk factors in frontline nurses under COVID-19 pandemic. Methods: This cross-sectional study was conducted from February 20, 2020, to March 20, 2020, and involved 562 frontline nurses. The effective response rate was 87.68%. After propensity score matched, there were 532 participants left. Extensive characteristics, including demographics, dietary habits, life-related factors, work-related factors, and psychological factors were collected based on a self-reported questionnaire. Specific scales measured the levels of sleep quality, physical activity, anxiety, perceived organization support and psychological capital. Adjusted odds ratios and 95% confidence intervals were determined by binary paired logistic regression. Results: Of the nurses enrolled in the study, 33.60% had anxiety. Five independent risk factors were identified for anxiety: poor sleep quality (OR=1.235), experienced major events (OR=1.653), lower resilience and optimism of psychological capital (OR=0.906, and OR=0.909) and no visiting friend constantly (OR=0.629). Conclusions: This study revealed a considerable high prevalence of anxiety in frontline nurses during the COVID-19 outbreak, and identified five risk factors, which were poor sleep quality, experienced major events, lower resilience and optimism of psychological capital, and no visiting friend constantly. Protecting mental health of nurses is important for COVID-19 pandemic control and their wellbeing. These findings enrich the existing theoretical model of anxiety and demonstrated a critical need for additional strategies that could address the mental health in frontline nurses for policymakers.

Effect of comprehensive nursing on the pain, anxiety and malnutrition of hepatitis B patients.

OBJECTIVE: To explore the effect of comprehensive nursing on pain, anxiety and malnutrition of hepatitis B patients. METHODS: Totally 100 cases of hepatitis B patients received treatment in our hospital from July 2017 to July 2018 were divided into a study group (64 cases) and a control group (36 cases) for comprehensive nursing and routine nursing. The liver function, nutritional indexes, cognition of relevant nursing knowledge, VAS score, HAMD and HAMA scores, QOL score, total effective rate and incidence of adverse reactions were detected. RESULTS: Compared with the control group, the study group had lower liver function indexes and higher nutrition indexes, suggesting that the liver function of patients in the study group recovered better. Besides, the VAS score of the study group was lower, suggesting better pain relief in the study group. Moreover, the study group had higher scores of related nursing knowledge and lower scores of HAMD and HAMA, indicating that the nursing method in the study group was more effective in reducing depression and anxiety. Higher QOL score, higher total effective rate and lower incidence of adverse reactions of the study group revealed that the nursing mode adopted in this group was better for the recovery of patients. CONCLUSION: Comprehensive nursing can effectively alleviate the pain of hepatitis B patients, relieve their anxiety and other negative emotions, and improve their malnutrition.

The prevalence and risk factors for depressive symptoms in frontline nurses under COVID-19 pandemic based on a large cross-sectional study using the propensity score-matched method.

INTRODUCTION: To explore the prevalence of depressive symptoms and the associated risk factors in frontline nurses under COVID-19 pandemic. METHODS: This cross-sectional study was conducted from February 20, 2020 to March 20, 2020 and involved 562 frontline nurses. The effective response rate was 87.68%. After propensity score matched, there were 498 participants left. Extensive characteristics, including demographics, dietary habits, life-related factors, work-related factors, and psychological factors were collected based on a self-reported questionnaire. Specific scales measured the levels of sleep quality, physical activity, depressive symptoms, perceived organization support and psychological capital. Adjusted odds ratios and 95% confidence intervals were determined by binary paired logistic regression. RESULTS: Of the nurses enrolled in the study, 50.90% had depressive symptoms. Three independent risk factors were identified: poor sleep quality (OR = 1.608, 95% CI: 1.384-1.896), lower optimism of psychological capital (OR = 0.879, 95% CI: 0.805-0.960) and no visiting friend constantly (OR = 0.513, 95% CI: 0.286-0.920). CONCLUSIONS: This study revealed a considerable high prevalence of depressive symptoms in frontline nurses during the COVID-19 outbreak, and identified three risk factors, which were poor sleep quality, lower optimism of psychological capital, and no visiting friend constantly. Protecting mental health of nurses is important for COVID-19 pandemic control and their wellbeing. These findings enrich the existing theoretical model of depression and demonstrated a critical need for additional strategies that could address the mental health in frontline nurses for policymakers.

A Convalescent of COVID-19 with RT-PCR Test Continues Positive in Stool.

BACKGROUND: Coronavirus disease (COVID-19) has affected more than 100 countries worldwide and the discharge criteria of patients with COVID-19 vary across different countries. In China, patients with two negative respiratory viral RNA tests taken at least one day apart can be discharged with no further quarantine required. Currently, PCR testing of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in fecal sample is not routinely performed. METHODS: We present a patient with COVID-19, whose respiratory swabs became negative but fecal sample remained positive for SARS-CoV-2 RNA. RESULTS: Stool sample collected on 27th of February was still positive for SARS-CoV-2 RNA, 24 days after the first negative respiratory swab. CONCLUSIONS: Based on the experience from the 2003 SARS epidemic, we recommend that fecal RNA testing of SARS-CoV-2 should be incorporated into the discharge criteria to minimize the risk of transmission from the gastrointestinal tract.

Activating transcription factor 2 (ATF2) negatively regulates the expression of antimicrobial peptide genes through tumor necrosis factor (TNF) in Macrobrachium nipponense.

Activating transcription factor 2 (ATF2), a member of the bZIP transcription factor family, is involved in multiple physiological and developmental processes, yet its role in the innate immunity remains unclear. In this study, two isoforms (named as MnATF2a and MnATF2b) of ATF2 gene were identified in Macrobrachium nipponense and were produced by exon skipping. The full length of MnATF2a is 2328 bp with an open reading frame of 2079 bp that encode 692 amino acids. MnATF2a has 237 bp nucleotides more than MnATF2b and the extra 237 bp is a complete exon. MnATF2a and MnATF2b proteins contain the same conserved and typical bZIP domain at the C-terminus. MnATF2a has 79 amino acids more than MnATF2b. MnATF2a and MnATF2b are widely distributed in a variety of immune tissues. After Vibrio parahaemolyticus and Staphylococcus aureus infection, the expression levels of MnATF2a and MnATF2b were significant up-regulated in the gills and stomach at 12 h. RNA interference analysis showed that knockdown of the total MnATF2 gene significantly inhibits the transcription of tumor necrosis factor (TNF) and promotes the expression of crustins (including Cru3, Cru4, and Cru7). Further study showed that knockdown of MnTNF evidently increase the expression of Cru3, Cru4, and Cru7. Our research indicates that ATF2 negatively regulate the expression of AMPs by regulating the transcription of TNF in M. nipponense. This study provides valuable information about the function of ATF2 family in the innate immunity in crustacean.

Immune activation by a multigene family of lectins with variable tandem repeats in oriental river prawn (Macrobrachium nipponense).

Genomic regions with repeated sequences are unstable and prone to rapid DNA diversification. However, the role of tandem repeats within the coding region is not fully characterized. Here, we have identified a new hypervariable C-type lectin gene family with different numbers of tandem repeats (Rlecs; R means repeat) in oriental river prawn (Macrobrachium nipponense). Two types of repeat units (33 or 30 bp) are identified in the second exon, and the number of repeat units vary from 1 to 9. Rlecs can be classified into 15 types through phylogenetic analysis. The amino acid sequences in the same type of Rlec are highly conservative outside the repeat regions. The main differences among the Rlec types are evident in exon 5. A variable number of tandem repeats in Rlecs may be produced by slip mispairing during gene replication. Alternative splicing contributes to the multiplicity of forms in this lectin gene family, and different types of Rlecs vary in terms of tissue distribution, expression quantity and response to bacterial challenge. These variations suggest that Rlecs have functional diversity. The results of experiments on sugar binding, microbial inhibition and clearance, regulation of antimicrobial peptide gene expression and prophenoloxidase activation indicate that the function of Rlecs with the motif of YRSKDD in innate immunity is enhanced when the number of tandem repeats increases. Our results suggest that Rlecs undergo gene expansion through gene duplication and alternative splicing, which ultimately leads to functional diversity.

Leucine-rich repeat-containing G-protein-coupled receptor 2 (LGR2) can regulate PO activity and AMP genes expression in Macrobrachium nipponense.

Leucine-rich repeat-containing G-protein-coupled receptors (LGRs) form a subfamily of the large superfamily of G-protein-coupled receptors. LGRs can be divided into three groups. LGR2 from Drosophila melanogaster is involved in cuticle tanning (melanization and sclerotization). In this study, one LGR2 (MnLGR2) was identified from Macrobrachium nipponense. MnLGR2 has an open reading frame of 4515 bp encoding a protein with 1504 amino acids. MnLGR2 is comprised of a 7-transmembrane domain, 12 leucine-rich repeats, and 5 low-complexity regions. The highest expression level of MnLGR2 was observed in gills. The expression levels of MnLGR2 in gills and stomach could be regulated by bacterial challenge. Knockdown of MnLGR2 upregulated the expression of anti-microbial peptide (AMP) genes. Further study indicated that inhibition of AMP expression by MnLGR2 was through inhibition of relish-mediated AMP expression. In addition to the negative regulation of AMP expression, MnLGR2 participated in positive regulation of phenol oxidase (PO) activity and expression of proPO activating pathway-related genes (proPO-activating factor and proPO-activating enzymes). Therefore, MnLGR2 plays an important role in prawn innate immunity.