Gut-lung crosstalk in pulmonary involvement with inflammatory bowel diseases

Pulmonary abnormalities,dysfunction or hyper-reactivity occurs in association with inflammatory bowel disease(IBD) more frequently than previously recognized.Emerging evidence suggests that subtle inflammation exists in the airways among IBD patients even in the absence of any bronchopulmonary symptoms,and with normal pulmonary functions. The pulmonary impairment is more pronounced in IBD patients with active disease than in those in remission. A growing number of case reports show that the IBD patients develop rapidly progressive respiratory symptoms after colectomy,with failure to isolate bacterial pathogens on repeated sputum culture,and often request oral corticosteroid therapy. All the above evidence indicates that the inflammatory changes in both the intestine and lung during IBD. Clinical or subclinical pulmonary inflammation accompanies the main inflammation of the bowel.Although there are clinical and epidemiological reports of chronic inflammation of the pulmonary and intestinal mucosa in IBD,the detailed mechanisms of pulmonaryintestinal crosstalk remain unknown. The lung has no anatomical connection with the main inflammatory site of the bowel. Why does the inflammatory process shift from the gastrointestinal tract to the airways? The clinical and subclinical pulmonary abnormalities,dysfunction,or hyper-reactivity among IBD patients need further evaluation. Here,we give an overview of the concordance between chronic inflammatory reactions in the airways and the gastrointestinal tract. A better understanding of the possible mechanism of the crosstalk among the distant organs will be beneficial in identifying therapeutic strategies for mucosal inflammatory diseases such as IBD and allergy.

Nanosecond pulsed electric feld interrupts the glycogen metabolism in hepatocellular carcinoma by modifying the osteopontin pathway

To the Editor:Hepatocellular carcinoma (HCC) is one of the most common lethal cancers and is the leading cause of cancer mortality worldwide [1].In recent years,developments in locoregional therapies have provided new options for the treatment of HCC,which are recommended by the international guidelines including the Barcelona Clinic Liver Cancer (BCLC) and the American Joint Committee on Cancer (AJCC)[2].

Characterization of the pheromone receptors in Mythimna loreyi reveals the differentiation of sex pheromone recognition in Mythimna species

Pheromone receptors(PRs)are key proteins in the molecular mechanism of pheromone recognition,and exploring the functional differentiation of PRs between closely related species helps to understand the evolution of moth mating systems.Pheromone components of the agricultural pest Mythimna loreyi have turned into(Z)-9-tetradecen-1-yl acetate(Z9-14:OAc),(Z)-7-dodecen-1-yl acetate(Z7-12:OAc),and(Z)-11-hexadecen-1-yl acetate,while the composition differs from that of M.separata in the genus Mythimna.To understand the molecular mechanism of pheromone recognition,we sequenced and analyzed antennal transcriptomes to identify 62 odorant receptor(OR)genes.The expression levels of all putative ORs were analyzed using differentially expressed gene analysis.Six candidate PRs were quantified and functionally characterized in the Xenopus oocytes system.MlorPR6 and MlorPR3 were determined to be the receptors of major and minor components Z9-14:OAc and Z7-12:OAc.MlorPR1 and female antennae(FA)-biased MlorPR5 both possessed the ability to detect pheromones of sympatric species,including(Z,E)-9,12-tetradecadien-1-ol,(Z)-9-tetradecen-1-ol,and(Z)-9-tetradecenal.Based on the comparison of PR functions between M.loreyi and M.separata,we analyzed the differentiation of pheromone recognition mechanisms during the evolution of the mating systems of 2 Mythimna species.

Functional characterization of sex pheromone receptors in Spodoptera frugiperda, S. exigua, and S. litura moths

Moths possess an extremely sensitive and diverse sex pheromone processing system,in which pheromone receptors(PRs)are essential to ensure communication between mating partners.Functional properties of some PRs are conserved among species,which is important for reproduction.However,functional differentiation has occurred in some homologous PR genes,which may drive species divergence.Here,using genome analysis,17 PR genes were identified from Spodoptera frugiperda,S.exigua,and S.litura,which belong to 6 homologous groups(odorant receptor[OR]6,11,13,16,56,and 62);of which 6 PR genes(OR6,OR11,OR13,OR16,OR56,and OR62)were identified in S.frugiperda and S.exigua,and 5 PR genes were identified in S.litura,excluding OR62.Using heterologous expression in Xenopus oocytes,we characterized the functions of PR orthologs including OR6,OR56,and OR62,which have not been clarified in previous studies.OR6 orthologs were specifically tuned to(Z,E)-9,12-tetradecadienyl acetate(Z9,E12-14:OAc),and OR62 orthologs were robustly tuned to Z7-12:OAc in S.frugiperda and S.exigua.The optimal ligand for OR56 was Z7-12:OAc in S.frugiperda,but responses were minimal in S.exigua and S.litura.In addition,SfruOR6 was male antennae-specific,whereas SfruOR56 and SfruOR62 were male antennae-biased.Our study further clarified the functional properties of PRs in 3 Spodoptera moth species,providing a comprehensive understanding of the mechanisms of intraspecific communication and interspecific isolation in Spodoptera.

Functional map of the macroglomerular complex of male Helicoverpaarmigera

The mechanism of sex pheromone reception in the male cotton bollworm Helicoverpa armigera has been extensively studied because it has become an important model system for understanding insect olfaction.However,the pathways of pheromone processing from the antenna to the primary olfactory center in H.armigera have not yet been clarified.Here,the physiology and morphology of male H.armigera olfactory sensory neurons(OSNs)were studied using single sensillum recording along with anterograde filling and intracellular recording with retrograde filling.OSNs localized in type A sensilla responded to the major pheromone component cis-11-hexadecenal,and the axonal terminals projected to the cumulus(Cu)of the macroglomerular complex(MGC).The OSNs in type B sensilla respondcd to the bchavioral antagonist cis-9-tetradecenal,and the axonal terminals projected to the dorsomedial anterior(DMA)unit of the MGC.In type C sensilla,there were 2 OSNs:one that responded to cis-9-tetradecenal and cis-11-hexadecenol with the axonal terminals projecting to the DMA,and another that responded to the secondary pheromone components cis-9-hexadecenal and cis-9-tetradecenal with the axonal terminals projecting to the dorsomedial posterior(DMP)unit of the MGC.Type A and type B sensilla also housed the secondary OSNs,which were silent neurons with axonal terminals projected to the glomerulus G49 and DMP.Overall,the neural pathways that carry information on attractiveness and aversiveness in response to female pheromone components in H.armigera exhibit distinct projections to the MGC units.

Interferon-Gamma Release Assay is Not Appropriate for the Diagnosis of Active Tuberculosis in High-Burden Tuberculosis Settings： A Retrospective Multicenter Investigation

Background：Interferon-gamma release assay （IGRA） has been used in latent tuberculosis （TB） infection and TB diagnosis,but the results from different high TB-endemic countries are different.The aim of this study was to investigate the value of IGRA in the diagnosis of active pulmonary TB （PTB） in China.Methods：We conducted a large-scale retrospective multicenter investigation to further evaluate the role of IGRA in the diagnosis of active PTB in high TB-epidemic populations and the factors affecting the performance of the assay.All patients who underwent valid T-SPOT.TB assays from December 2012 to November 2015 in six large-scale specialized TB hospitals in China and met the study criteria were retrospectively evaluated.Patients were divided into three groups：Group 1,sputum culture-positive PTB patients,confirmed by positive Mycobacterium tuberculosis sputum culture;Group 2,sputum culture-negative PTB patients;and Group 3,non-TB respiratory diseases.The medical records of all patients were collected.Chi-square tests and Fisher＇s exact test were used to compare categorical data.Multivariable logistic analyses were performed to evaluate the relationship between the results of T-SPOT in TB patients and other factors.Results：A total of 3082 patients for whom complete information was available were included in the investigation,including 905 sputum culture-positive PTB cases,914 sputum cultmre-negative PTB cases,and 1263 non-TB respiratory disease cases.The positive rate of T-SPOT.TB was 93.3% in the culture-positive PTB group and 86.1% in the culture-negative PTB group.In the non-PTB group,the positive rate of T-SPOT.TB was 43.6%.The positive rate of T-SPOT.TB in the culture-positive PTB group was significantly higher than that in the culture-negative PTB group （x2 =25.118,P 〈 0.01）,which in turn was significantly higher than that in the non-TB group （x2 =566.l 16,P 〈 0.01）.The overall results were as follows：sensitivity,89.7%;specificity,56.37%;positive predictive value,74.75%;negative predictive value,79.11%;and accuracy,76.02%.Conclusions：High false-positive rates of T-SPOT.TB assays in the non-TB group limit the usefulness as a single test to diagnose active TB in China.We highly recommend that IGRAs not be used for the diagnosis of active TB in high-burden TB settings.

Identification and functional characterization of an odorant receptor in pea aphid,Acyrthosiphon pisum

The sensitive olfactory system is necessary for survival of insects.Odorant receptors (ORs)are located on the dendrites of olfactory receptor neurons and play a critical role in odor detection.Insect ORs are functionally analyzed via heterologous expression in a Xenopus oocyte system using a two-electrode voltage-clamp (TEVC)electrophysiological recording.Here,we have identified a novel OR in the pea aphid,Acyrthosiphon pisum,then we cloned and named it ApisOR4.We analyzed the ApisOR4 tissue expression patterns and found expression only in antennae tissues.Further functional analysis using TEVC revealed that ApisOR4 is broadly tuned to eight volatiles,which elicit electrophysiological response in pea aphid antennae.This study provides an initial functional analysis of aphid ORs and identifies candidate volatiles to be used in developing new strategies for aphid control.

A de novo transcriptomic analysis to reveal functional genes in Apolygus lucorum

The widespread planting of genetically engineered cotton producing the CrylAc toxin has led to significantly reduced pesticide applications since 1997. However, conse- quently, the number of green mirid bugs （GMB）, Apolygus lucorum （Meyer-Diir） has in- creased. So far the GMB, instead of the cotton bollworm Helicoverpa armigera （Hiibner）, has become the major pest in the transgenic Bt cotton field and has influenced cotton yield. Disproportionately, only a few studies on GMB at molecular level have been re- ported. Libraries from both third instar nymphs and adults were sequenced using Illumina technology, producing more than 106 million short reads and assembled into 63 029 uni- genes of mean length 597 nt and N50 813 nt, ranging from 300 nt to 9771 nt. BLASTx analysis against Nr, Swissprot, GO and COG was performed to annotate these unigenes. As a result, 26 478 unigenes （42.01%） matched to known proteins and 107 immune-related, 320 digestive-related and 53 metamorphosis-related genes were detected in these annotated unigenes. Additionally, we profiled gene expression using mapping based differentially expressed genes （DEGs） strategy between the two developmental stages： nymph and adult. The results demonstrated that thousands of genes were significantly differentially ex- pressed at different developmental stages. The transcriptome and gene expression data provided comprehensive and global gene resources of GMB. This transcriptome would improve our understanding of the molecular mechanisms of various underlying biological characteristics, including development, digestion and immunity in GMB. Therefore, these findings could help elucidate the intrinsic factors of the GMB resurgence, offering novel pest management targets for future transgenic cotton breeding.

Identification and localization of two sensory neuron membrane proteins from Spodoptera fitura （Lepidoptera： Noctuidae）

Sensory neuron membrane proteins （SNMPs）, which are located on the dendritic membrane of olfactory sensory neurons （OSNs）, are proposed to be associated with odor reception in insects. Recent studies have demonstrated that SNMP1 is essential for electrophysiological responses of OSNs to the sex pheromone, cis-vaccenyl acetate （cVA） in Drosophila melanogaster. To investigate the function ofLepidoptera SNMPs, we cloned two SNMP genes, SlituSNMP1 and SltiuSNMP2, from Spodoptera litura （Lepidoptera： Noctuidae）. Sequence alignment and phylogenetic analysis showed that both genes bear the general characteristics of SNMPs, including six conserved cysteine residues and two transmembrane domains. Further expression profile experiments showed that SIituSNMP1 is mainly expressed in the antenna, while SlituSNMP2 is broadly expressed in various tissues. By in situ hybridization experiments, it was found that SlituSNMP1 expressing cells are surrounded by the SlituSNMP2 expressing cells in the pheromone sensitive sensilla, suggesting different fimctions ＆the two SNMPs in insect olfaction.

Functional characterization of pheromone receptor candidates in codling moth Cydia pomonella(Lepidoptera:Tortricidae)

Sex pheromones serve a critical role in Lepidopterans finding mates.Male moths perceive and react to sex pheromones emitted by conspecific females through a delicate pheromone communication system.Pheromone receptors(PRs)are the key sensory elements at the beginning of that process.The codling moth(Cydia ponwonella)is an important pome fruit pest globally and a serious invasive species in China.Pheromonebased techniques have been used successfully in monitoring and controlling this species.We conducted ribonucleic acid sequencing analysis of the codling moth antennal transcriptome and identified 66 odorant receptors(ORs)in a population from Xinjiang province,China,of which 14 were PRs,including two novel PRs(CpomOR2e and CpomOR73).Four PRs that contain full-length open reading frames(CpomORl,OR2a,OR5,OR7)and four PRs with ligands that have not been reported previously(CpomORl,OR2a,OR5,OR7)were selected to deorphanize in the heterologous Xenopus oocyte expression system.Specifically,we found that CpomOR2a and CpomOR5 responded to(£,£)-8,10-dodecadien-l-yl acetate(codlemone acetate).Furthermore,CpomOR5(EC50=1.379 x 10-8 mol/L)was much more sensitive to codlemone acetate than CpomOR2a(EC50=1.663×10^(-6) mol/L).Since codlemone acetate is an important component of C.pomonella sex pheromone,our results improve the current understanding of pheromone communication in codling moths and will be helpful for the development of pest management strategies.

The general odorant receptor GmolOR9 from Grapholita molesta (Lepidoptera: Tortricidae) is mainly tuned to eight host-plant volatiles

Grapholita molesta is one of the most destructive fruit pests distributed worldwide.Odorant receptors(ORs)located on the dendritic membrane of chemosensory neurons are deemed to be key molecules for sensing exogenous chemical signals.In this study,GmolOR9,a general OR from G.molesta,was functionally characterized.Quantitative real-time polymerase chain reaction revealed that GniolOR9 was more highly expressed in adults than in other stages,including eggs,larvae,and pupae.GmolOR9 expression was highly significantly more in the antennae of females than in those of males,and the highest level occurred in the antennae of 3-day-old female adults.GmolOR9 was broadly tuned to eight of 47 odorant components tested including(Z)-3-hexenyl acetate,butyl propionate,ethyl hexanoate,ethyl heptanoate,1-hexanol,(Z)-3-hexenol,2-ethyl-1-hexanol,and linalool,by in vitro heterologous expression.Furthermore,electroantennogram responses indicated that the efTects of dsOR9-injected females to(Z)-3-hexenyl acetate dramatically decreased.These results suggested that GmolOR9 might be involved in detecting hostplant volatiles.Moreover,(Z)-3-hexenyl acetate might serve as a potential attractant for the biological control of G.molesta.

Neuronal architecture and functional mapping of the taste center of larval Helicoverpa armigera(Lepidoptera:Noctuidae)

The sense of taste plays a crucial role in herbivorous insects by discriminating nutrients from complex plant metabolic compounds.The peripheral coding of taste has been thoroughly studied in many insect species,but the central gustatory pathways are poorly described.In the present study,we characterized single neurons in the gnathal ganglion of Helicoverpa armigera larvae using the intracellular recording/staining technique.We identified different types of neurons,including sensory neurons,interneurons,and motor neurons.The morphologies of these neurons were largely diverse and their arborizations seemingly covered the whole gnathal ganglion.The representation of the single neurons responding to the relevant stimuli of sweet and bitter cues showed no distinct patterns in the gnathal ganglion.We postulate that taste signals may be processed in a manner consistent with the principle of population coding in the gnathal ganglion of H.armigera larvae.

Ionotropic receptor 8a is involved in the attraction of Helicoverpa armigera to acetic acid

Ionotropic receptors(IRs)were first found in Drosophila melanogaster,and derive from ionotropic glutamate receptors(iGluRs),which are implicated in detecting acids,ammonia,amine,temperature and humidity.Although IRs are involved in sensing acid odors in a few insects,such as D.melanogaster,Aedes aegypti,and Manduca sexta,the function of IRs in Helicoverpa armigera is still unknown.IR8a was confirmed to be a co-receptor associated with acid detection.From the results of phylogenetic analysis,HarmIR8a displayed high similarity compared to homologs in D.melanogaster,M.sexta,and A.aegypti,suggesting that HarmIR8a might have a consistent function as a co-receptor for acid detection.In this study,clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)-mediated genome editing was implemented to knockout HarmIR8a for in vivo functional analysis.Electrophysiological and behavioral assays were performed to compare the differences between HarmIR8a knockout mutants and wild type individuals.From electroantennogram(EAG)analysis,we found that wild type H.armigera adults could detect short-chain carboxylic acids.In addition,wind tunnel experiments showed that 1%acetic acid attracted wild type H.armigera adults.However,acid sensing and attraction were reduced or abolished in the HarmIR8a knockout mutants.Our data suggest that HarmIR8a is important for H.armigera to detect short-chain carboxylic acids and mediate attraction behavior to acetic acid.