Neutrophil extracellular traps mediate cardiomyocyte ferroptosis via the Hippo-Yap pathway to exacerbate doxorubicin-induced cardiotoxicity.

Doxorubicin-induced cardiotoxicity (DIC), which is a cardiovascular complication, has become the foremost determinant of decreased quality of life and mortality among survivors of malignant tumors, in addition to recurrence and metastasis. The limited ability to accurately predict the occurrence and severity of doxorubicin-induced injury has greatly hindered the prevention of DIC, but reducing the dose to mitigate side effects may compromise the effective treatment of primary malignancies. This has posed a longstanding clinical challenge for oncologists and cardiologists. Ferroptosis in cardiomyocytes has been shown to be a pivotal mechanism underlying cardiac dysfunction in DIC. Ferroptosis is influenced by multiple factors. The innate immune response, as exemplified by neutrophil extracellular traps (NETs), may play a significant role in the regulation of ferroptosis. Therefore, the objective of this study was to investigate the involvement of NETs in doxorubicin-induced cardiomyocyte ferroptosis and elucidate their regulatory role. This study confirmed the presence of NETs in DIC in vivo. Furthermore, we demonstrated that depleting neutrophils effectively reduced the occurrence of doxorubicin-induced ferroptosis and myocardial injury in DIC. Additionally, our findings showed the pivotal role of high mobility group box 1 (HMGB1) as a critical molecule implicated in DIC and emphasized its involvement in the modulation of ferroptosis subsequent to NETs inhibition. Mechanistically, we obtained preliminary evidence suggesting that doxorubicin-induced NETs could modulate yes-associated protein (YAP) activity by releasing HMGB1, which subsequently bound to toll like receptor 4 (TLR4) on the cardiomyocyte membrane, thereby influencing cardiomyocyte ferroptosis in vitro. Our findings suggest that doxorubicin-induced NETs modulate cardiomyocyte ferroptosis via the HMGB1/TLR4/YAP axis, thereby contributing to myocardial injury. This study offers a novel approach for preventing and alleviating DIC by targeting alterations in the immune microenvironment.

Protective effect of low-intensity pulsed ultrasound on immune checkpoint inhibitor-related myocarditis via fine-tuning CD4+ T-cell differentiation.

PURPOSE: Immune checkpoint inhibitors (ICIs) have transformed traditional cancer treatments. Specifically, ICI-related myocarditis is an immune-related adverse event (irAE) with high mortality. ICIs activate CD4+ T-lymphocyte reprogramming, causing an imbalance between Th17 and Treg cell differentiation, ultimately leading to myocardial inflammatory damage. Low-intensity pulsed ultrasound (LIPUS) can limit inflammatory responses, with positive therapeutic effects across various cardiovascular inflammatory diseases; however, its role in the pathogenesis of ICI-related myocarditis and CD4+ T-cell dysfunction remains unclear. Accordingly, this study investigated whether LIPUS can alleviate ICI-related myocarditis inflammatory damage and, if so, aimed to elucidate the beneficial effects of LIPUS and its underlying molecular mechanisms. METHODS: An in vivo model of ICI-related myocarditis was obtained by intraperitonially injecting male A/J mice with an InVivoPlus anti-mouse PD-1 inhibitor. LIPUS treatment was performed via an ultrasound-guided application to the heart via the chest wall. The echocardiographic parameters were observed and cardiac function was assessed using an in vivo imaging system. The expression of core components of the HIPPO pathway was analyzed via western blotting. RESULTS: LIPUS treatment reduced cardiac immune responses and inflammatory cardiac injury. Further, LIPUS treatment alleviated the inflammatory response in mice with ICI-related myocarditis. Mechanistically, in the HIPPO pathway, the activation of Mst1-TAZ axis improved autoimmune inflammation by altering the interaction between the transcription factors FOXP3 and RORγt and regulating the differentiation of Treg and Th17 cells. CONCLUSION: LIPUS therapy was shown to reduce ICI-related myocarditis inflammatory damage and improve cardiac function, representing an exciting finding for irAEs treatment.

The ecdysis triggering hormone system is essential for reproductive success in Mythimna separata (Walker).

Ecdysis triggering hormone (ETH) was originally discovered as a key hormone that regulates insect moulting via binding to its receptor, ETH receptor (ETHR). However, the precise role of ETH in moth reproduction remains to be explored in detail. ETH function was verified in vivo using Mythimna separata (Walker), an important cereal crop pest. RT-qPCR analysis revealed that transcriptional expression profiles of MsepETH showed evident sexual dimorphism in the adult stage. MsepETH expression increased in the females on day 3 and persisted thereafter till day 7, consistent with female ovarian maturation, and was merely detectable in males. Meanwhile, MsepETH expression levels were significantly higher in the trachea than in other tissues. MsepETHR-A and MsepETHR-B were expressed in both sexes and were significantly higher in the antennae than in other tissues. MsepETH and MsepETHR knockdown in females by RNA interference significantly reduced the expression of MsepETH, MsepETHR-A, MsepETHR-B, MsepJHAMT, and MsepVG, which delayed egg-laying and significantly reduced egg production. RNAi 20-hydroxyecdysone (20E) receptor (EcR) decreased MsepETH expression whereas injecting 20E restored egg production that had been disrupted by MsepETH interference. Meanwhile, RNAi juvenile hormone (JH) methoprene tolerant protein (Met) also decreased MsepETH expression and smearing JH analog methoprene (Meth) restored egg production. In conclusion, the reproduction roles of ETH, JH, and 20E were investigated in M. separata. These findings will lay the foundation for future research to develop an antagonist that reduces female reproduction and control strategies for pest insects.

Morph-specific fitness throughout the life cycle of the grain aphid, nonhost-alternating, holocyclic Sitobion avenae (Hemiptera: Aphididae).

Aphids exhibit seasonally alternating asexual and sexual reproductive modes. Different morphs are produced throughout the life cycle. To evaluate morph-specific fitness during reproductive switching, holocyclic Sitobion avenae were induced continuously under short light conditions, and development and reproduction were compared in each morph. Seven morphs, including apterous and alate virginoparae, apterous and alate sexuparae, oviparae, males, and fundatrices, were produced during the life cycle. The greatest proportions of sexuparae, oviparae, males, and virginoparae were in the G1, G2, G3, and G4 generations, respectively. Regardless of asexual or sexual morphs, alate morphs exhibited a marked delay in age at maturity compared with that of apterous morphs. Among the alate morphs, males had the longest age at maturity, followed by sexuparae and virginoparae. Among the apterous morphs, sexuparae were older at maturity than the fundatrices, virginoparae, and oviparae. The nymphs of each morph had equal survival potentials. For the same wing morphs, apterous sexuparae and oviparae exhibited substantial delays in the pre-reproductive period and considerable reductions in fecundity, compared with those of apterous virginoparae and fundatrices, whereas alate sexuparae and alate virginoparae had similar fecundity. The seven morphs exhibited Deevey I survivorship throughout the life cycle. These results suggest that sexual production, particularly in males, has short-term development and reproduction costs. The coexistence of sexual and asexual morphs in sexuparae offspring may be regarded as an adaptive strategy for limiting the risk of low fitness in winter.

Melatonin alleviates doxorubicin-induced mitochondrial oxidative damage and ferroptosis in cardiomyocytes by regulating YAP expression.

Doxorubicin (Dox) is a high-efficiency agent for cancer therapy. However, it causes cardiotoxicity which limits its clinical application. Despite more efforts has been made to seek protective decisions, unfortunately, the poor prognosis suggests the need for new treatments. As a powerful mitochondrial antioxidant, melatonin (Mel) has been found to confer cardioprotection against various cardiovascular diseases. Currently, the mechanism through which Mel confers protection is not well understood. In this study, we established a Dox-induced cardiotoxicity model in H9c2 cardiomyocytes, zebrafish, and SD rats to explore the mechanism by which Mel alleviates Dox-induced cardiotoxicity. In vivo and in vitro experiments showed that Dox significantly decreased the viability of H9c2 cells, induced apoptosis, myocardial injury, and effectively up-regulated the expression of p-YAP but down-regulated the expression of YAP. Furthermore, we found that Dox significantly up-regulated the expression of ferroptosis-associated protein ACSL4 and down-regulated expression of GPX4. Interestingly, these effects of Dox were reversed following treatment with Mel, indicating that ferroptosis mediated the protective effects of Mel against Dox-induced cardiomyocyte injury. Furthermore, we used YAP-siRNA in vitro and verteporfin (Ver) in vivo to down-regulate the expression level of YAP. The results showed that YAP down-regulation abolished the protective effects of Mel including apoptosis, mitochondrial lipid peroxidation, and ferroptosis. Collectively, these results show that Mel regulates ferroptosis by modulating YAP expression to counteract Dox-induced cardiotoxicity.

Polyphenism in Antennal Sensilla Among Different Adult Morphs of Nonhost-Alternating, Holocyclic Sitobion avenae (Hemiptera: Aphididae).

Aphids, mainly distributed in temperate zones, exhibit seasonal generation-alternating phenomena. Across the life cycle, different morphs are produced. Sitobion avenae (Fabricius 1775) is a major pest of wheat worldwide. To elucidate olfactory perception of morph-specific behavior across their life cycle, we investigated antennal sensilla among seven morphs using scanning electron microscopy. Trichoid, placoid, coeloconic, and campaniform sensilla were identified. Trichoid sensilla, big multiporous placoid sensilla (primary rhinarium), a group of sensilla (primary rhinaria), and campaniform sensilla showed similar distribution and resemblance among morphs, whereas small multiporous placoid sensilla (secondary rhinaria) exhibited obvious differences. Compared to apterous morphs, alate morphs possessed a greater abundance of secondary rhinaria, with the greatest found in males on antennal segments III-V. Alate virginoparae and alate sexuparae ranged from six to fourteen rhinaria on antennal segment III. Fundatrices, apterous virginoparae and apterous sexuparae only had one or two secondary rhinaria on antennal segment III while they disappeared in oviparae. Secondary rhinaria, lying in a cuticle cavity, are convex or concave in their central part. In males, both forms were present, with a greater proportion of convex form than that of the concave form. Fundatrices and virginoparae had the convex form while sexuparae had the concave form. Polyphenism of secondary rhinaria might suggest their association with the olfactory functions of morph-specific behavior. These results have improved our understanding of the adaptive evolution of the antennal sensilla in nonhost-alternating, holocyclic aphids.

Demography of Mythimna separata (Lepidoptera: Noctuidae) at outdoor fluctuating temperatures.

The oriental armyworm Mythimna separata (Walker) (Lepidoptera: Noctuidae) is a major migratory pest of cereal crops in East Asia, South Asia and Australia. To comprehensively understand the ecological tolerance of M. separata, we collected life table data of individuals from four consecutive generations reared under outdoor natural fluctuating temperatures from 15 April to 17 October 2018 in Yangling, Shaanxi, China. The results showed that the immature stage in early summer and summer were shorter than in spring and autumn. High mortality in late larval instar and pupal stages was observed in the summer generation. The adult pre-oviposition period in autumn was longer than the other seasons. The population in the earlier two seasons had heavier pupae and higher fecundity than the population in the latter two seasons. The intrinsic rate of increase and the finite rate of increase was the highest in early summer (r = 0.1292 day-1, λ = 1.1391 day-1), followed by spring (r = 0.1102 day-1, λ = 1.1165 day-1), and was the lowest in summer (r = 0.0281 day-1, λ = 1.0293 day-1). The results of this study would be useful to predict the population dynamics of M. separata and deepen our standing of the adaptiveness of this migratory pest in natural fluctuating ambient environments.

Mucosal immune response of Nile tilapia Oreochromis niloticus during Gyrodactylus cichlidarum infection.

Monogenean Gyrodactylus cichlidarum can cause severe mortality of Nile tilapia (Oreochromis niloticus) fry. To date, reports about mucosal immunity of O. niloticus against this parasite have been rare. In order to explore the mucosal immunity of Nile tilapia against G. cichlidarum infection, the expressions of six adaptive immune-related genes and the contents of specific immunoglobulin IgM and IgT in the skin-associated lymphoid tissues (SALT) were dynamically analyzed after primary and secondary infections. The abundances of G. cichlidarum on the hosts after secondary infection were lower than those after primary parasite infection, which implied that hosts could initiate immune protection against G. cichlidarum reinfection to some degree. The transcription levels of TCR-ß and CD4 genes in the skin tissue were significantly up-regulated after primary G. cichlidarum infection, while genes pIgR and IgT were only detected with significant up-regulations during secondary infection. With the exception of pIgR, which had remarkably higher expression in the fish with low parasite loads, all other genes studied tended to have higher mRNA level in the fish with higher parasite loads. The specific IgM content in the skin mucus increased significantly on the 2nd day after the primary exposure, higher than the corresponding value during the secondary exposure, and had significantly positive correlation with the parasite loads during the first parasite infection. These results manifested that acquired immune responses in the SALT of Nile tilapia participated in the resistance against G. cichlidarum infection, underscoring the involvement of mucosal immunity in fish against monogenean infection, and suggesting potential prophylactic treatment of gyrodactylid disease of tilapia.

Expression of immune-related genes of Nile tilapia Oreochromis niloticus after Gyrodactylus cichlidarum and Cichlidogyrus sclerosus infections demonstrating immunosupression in coinfection.

Gyrodactylus cichlidarum and Cichlidogyrus sclerosus, two monogenean ectoparasite species commonly found on the body surface and gills of Nile tilapia (Oreochromis niloticus) respectively, inflicted considerable economic losses in intensive tilapia farming. In order to explore the immune response of tilapia against these two species of monogeneans, expression patterns of five immune-related genes were studied after singular G. cichlidarum or C. sclerosus infection and their coinfection. The transcription levels of IL-1ß were up-regulated in the skin after G. cichlidarum infection, reaching a peak at day 5 PI, and in the gills after C. sclerosus infection (peaking at day 8 PI), with significant elevation only detected in the gills after high-dose C. sclerosus infection. A trend favoring increased gill TNF-α expression at day 8 PI of C. sclerosus infection was statistically significant only in the low-dose infection group. TNF-α expression in the skin did not change significantly after G. cichlidarum infection. TGF-ß had extremely up-regulated expressions in the gills at day 8 PI after both high- and low-dose C. sclerosus infections, but its significantly promoted expression in the skin was observed only after infection of high-dose G. cichlidarum. Significantly increased expressions of HSP70 and COX-2 in the skin were detected after high-dose G. cichlidarum infections. In comparison to singular infection with either G. cichlidarum or C. sclerosus, concurrent infection resulted in significantly advanced expression of TGF-ß in both skin and gills, and lower expressions at day 8 PI, and similar patterns were observed in the expression of IL-1ß and TNF-α in the gills. G. cichlidarum infection on the body surface significantly down-regulated the expressions of TNF-α, TGF-ß and COX-2 in the gills. In addition, the intensity of G. cichlidarum was significantly positively correlated with that of C. sclerosus (correlation index 0.922, p = 0.000) at day 2 PI under concurrent infection. These results contribute to the understanding of mucosal immunity of fish against monogenean infection, particularly when two monogenean species infect concurrently.

Degradation of sex pheromone and plant volatile components by an antennal glutathione S-transferase in the oriental fruit moth,Grapholita molesta Busck (Lepidoptera: Tortricidae).

Insect antennae have a primary function of perceiving and discerning odorant molecules including sex pheromones and host plant volatiles. The assumption that genes highly expressed in the antennae may have an olfactory-related role associated with signal transduction. Here, one delta subfamily glutathione S-transferase (GST) gene (GmolGSTD1) was obtained from an antennal transcriptome of Grapholita molesta. Quantitative real-time polymerase chain reaction results revealed that GmolGSTD1 was mainly expressed in antennae and the expression levels were significantly higher in female antennae than in male antennae. The recombinant GmolGSTD1 (rGmolGSTD1) showed glutathione-conjugating activity toward 1-chloro-2,4-dinitrobenzene (CDNB) as substrates. The pH range for optimal rGmolGSTD1 enzyme activity was 6.0-6.5, and rGmolGSTD1 enzyme activity had maximal peaks at 35-40°C. Spectrophotometric analysis indicated that insecticides had weak inhibitory effects on the activity of rGmolGSTD1 with the inhibitory rates of 28.82% for chlorpyrifos, 22.27% for lambda-cyhalothrin, 18.07% for bifenthrin, 20.42% for acetamiprid, 17.57% for thiamethoxam, 25.67% for metaflumizone, 27.43% for abamectin, and 7.24% for chlorbenzuron. rGmolGSTD1 exhibited high degradation activity to the sex pheromone component (Z)-8-dodecenyl alcohol and the host plant volatile butyl hexanoate with the degradation efficiency of 75.01% and 48.54%, respectively. We speculate that GmolGSTD1 works in inactivating odorant molecules and maintaining sensitivity to olfactory communication of G. molesta.

CLONING, EXPRESSION, AND FUNCTIONAL ANALYSIS OF THREE ODORANT-BINDING PROTEINS OF THE ORIENTAL FRUIT MOTH, Grapholita molesta (BUSCK) (LEPIDOPTERA: TORTRICIDAE).

Odorant-binding proteins (OBPs) act in insect olfactory processes. OBPs are expressed in the olfactory organs and serve in binding and transport of hydrophobic odorants through the sensillum lymph to olfactory receptor neurons within the antennal sensilla. In this study, three OBP genes were cloned from the antennal transcriptome database of Grapholita molesta via reverse-transcription PCR. Recombinant GmolOBPs (rGmolOBPs) were expressed in a prokaryotic expression system and enriched via Ni ion affinity chromatography. The binding properties of the three rGmolOBPs to four sex pheromones and 30 host-plant volatiles were investigated in fluorescence ligand-binding assays. The results demonstrated that rGmolOBP8, rGmolOBP11, and rGmolOBP15 exhibited high binding affinities with the major sex pheromone components (E)-8-dodecenyl acetate, (Z)-8-dodecenyl alcohol, and dodecanol. The volatiles emitted from peach and pear, decanal, butyl hexanoate, and α-ocimene, also showed binding affinities to rGmolOBP8 and rGmolOBP11. Hexanal, heptanal, and α-pinene showed strong binding affinities to rGmolOBP15. Results of the electrophysiological recording experiments and previous behavior bioassays indicated that adult insects had strong electroantennogram and behavioral responses toward butyl hexanoate, hexanal, and heptanal. We infer that the GmolOBP8 and GmolOBP11 have dual functions in perception and recognition of host-plant volatiles and sex pheromones, while GmolOBP15 was mainly involved in plant volatile odorants' perception.

Effect of short neuropeptide F signaling on larval feeding in Mythimna separata.

Mythimna separata is a notorious phytophagous pest which poses serious threats to cereal crops owing to the gluttony of the larvae. Because short neuropeptide F (sNPF) and its receptor sNPFR are involved in a diversity of physiological functions, especially in functions related to feeding in insects, it is a molecular target for pest control. Herein, an sNPF and 2 sNPFRs were identified and cloned from M. separata. Bioinformatics analysis revealed that the sNPF and its receptors had a highly conserved RLRFamide C-terminus and 7 transmembrane domains, respectively. The sNPF and its receptor genes were distributed across larval periods and tissues, but 2 receptors had distinct expression patterns. The starvation-induced assay elucidated that sNPF and sNPFR expression levels were downregulated under food deprivation and recovered with subsequent re-feeding. RNA interference knockdown of sNPF, sNPFR1, and sNPFR2 by injection of double-stranded RNA into larvae not only suppressed food consumption and increased body size and weight, but also led to decrease of glycogen and total lipid contents, and increase of trehalose compared with double-stranded green fluorescent protein injection. Furthermore, molecular docking was performed on the interaction mode between sNPFR protein and its ligand sNPF based on the 3-dimensional models constructed by AlphaFold; the results indicated that both receptors were presumably activated by the mature peptide sNPF-2. These results revealed that sNPF signaling played a considerably vital role in the feeding regulation of M. separata and represents a potential control target for this pest.

Identification of the neuropeptide gene family and feeding regulation by neuropeptide Y in Mythimna separata (Lepidoptera: Noctuidae).

Mythimna separata is a crucial agricultural pest with polyphagous characteristic. Neuropeptide signaling, especially neuropeptide Y (NPY) involved in feeding regulation, is considered as a potential pest control target. In this study, we identified 50 neuropeptides including NPY of M. separata using transcriptome sequencing. Bioinformatics analysis showed that NPY had a conserved carboxyterminal motif of RGRYamide and shared very high homology across lepidopteran insects. Spatio-temporal expression profile articulated that NPY was mainly expressed in the larval stage, and with the highest expression in larval midgut and adult alimentary canal. RNAi knockdown by the injection of dsNPY into larvae significantly inhibited food uptake and body weight, delayed developmental duration, and also caused increase of trehalose and decrease of glycogen and total lipid compared to dsGFP. This study identified nearly the entire neuropeptide gene family of M. separata and further indicated that NPY signaling pathway might played a vital role in the feeding regulation and related to energy metabolism. Our results not only laid a preliminary foundation for functional studies on neuropeptide of M. separata, but also provided a potential molecular target for future pesticide development.

Deep learning-based classification of breast lesions using dynamic ultrasound video.

PURPOSE: We intended to develop a deep-learning-based classification model based on breast ultrasound dynamic video, then evaluate its diagnostic performance in comparison with the classic model based on ultrasound static image and that of different radiologists. METHOD: We collected 1000 breast lesions from 888 patients from May 2020 to December 2021. Each lesion contained two static images and two dynamic videos. We divided these lesions randomly into training, validation, and test sets by the ratio of 7:2:1. Two deep learning (DL) models, namely DL-video and DL-image, were developed based on 3D Resnet-50 and 2D Resnet-50 using 2000 dynamic videos and 2000 static images, respectively. Lesions in the test set were evaluated to compare the diagnostic performance of two models and six radiologists with different seniority. RESULTS: The area under the curve of the DL-video model was significantly higher than those of the DL-image model (0.969 vs. 0.925, P = 0.0172) and six radiologists (0.969 vs. 0.779-0.912, P < 0.05). All radiologists performed better when evaluating the dynamic videos compared to the static images. Furthermore, radiologists performed better with increased seniority both in reading images and videos. CONCLUSIONS: The DL-video model can discern more detailed spatial and temporal information for accurate classification of breast lesions than the conventional DL-image model and radiologists, and its clinical application can further improve the diagnosis of breast cancer.

Construction and validation of a nomogram prediction model for axillary lymph node metastasis of cT1 invasive breast cancer.

OBJECTIVE: Based on the ultrasonic characteristics of the breast mass and axillary lymph nodes as well as the clinicopathological information, a model was developed for predicting axillary lymph node metastasis in cT1 breast cancer, and relevant features associated with axillary lymph node metastasis were identified. METHODS: Our retrospective study included 808 patients with cT1 invasive breast cancer treated at the Second Affiliated Hospital and the Cancer Hospital Affiliated with Harbin Medical University from February 2012 to August 2021 (250 cases in the positive axillary lymph node group and 558 cases in the negative axillary lymph node group). We allocated 564 cases to the training set and 244 cases to the verification set. R software was used to compare clinicopathological data and ultrasonic features between the two groups. Based on the results of multivariate logistic regression analysis, a nomogram prediction model was developed and verified for axillary lymph node metastasis of cT1 breast cancer. RESULTS: Univariate and multivariate logistic regression analysis indicated that palpable lymph nodes (P = 0.003), tumor location (P = 0.010), marginal contour (P < 0.001), microcalcification (P = 0.010), surrounding tissue invasion (P = 0.046), ultrasonic detection of lymph nodes (P = 0.001), cortical thickness (P < 0.001) and E-cadherin (P < 0.001) are independently associated with axillary lymph node metastasis. Using these features, a nomogram was developed for axillary lymph node metastasis. The training set had an area under the curve of 0.869, while the validation set had an area under the curve of 0.820. Based on the calibration curve, the model predicted axillary lymph node metastases were in good agreement with reality (P > 0.05). Nomogram's net benefit was good based on decision curve analysis. CONCLUSION: The nomogram developed in this study has a high negative predictive value for axillary lymph node metastasis in invasive cT1 breast c ancer. Patients with no axillary lymph node metastases can be accurately screened using this nomogram, potentially allowing this group of patients to avoid invasive surgery.

Metabolic engineering of a 1,8-cineole synthase enhances aphid repellence and increases trichome density in transgenic tobacco (Nicotiana tabacum L.).

BACKGROUND: The green peach aphid (Myzus persicae Sulzer) is a harmful agricultural pest that causes severe crop damage by directly feeding or indirectly vectoring viruses. 1,8-cineole synthase (CINS) is a multiproduct enzyme that synthesizes monoterpenes, with 1,8-cineole dominating the volatile organic compound profile. However, the relationship between aphid preference and CINS remains elusive. RESULTS: Here, we present evidence that SoCINS, a protein from garden sage (Salvia officinalis), enhanced aphid repellence and increased trichome density in transgenic tobacco. Our results demonstrated that overexpression of SoCINS (SoCINS-OE) led to the emission of 1,8-cineole at a level of up to 181.5 ng per g fresh leaf. Subcellular localization assay showed that SoCINS localized to chloroplasts. A Y-tube olfactometer assay and free-choice assays revealed that SoCINS-OE plants had a repellent effect on aphids, without incurring developmental or fecundity-related penalties. Intriguingly, the SoCINS-OE plants displayed an altered trichome morphology, showing increases in trichome density and in the relative proportion of glandular trichomes, as well as enlarged glandular cells. We also found that SoCINS-OE plants had significantly higher jasmonic acid (JA) levels than wild-type plants. Furthermore, application of 1,8-cineole elicited increased JA content and trichome density. CONCLUSION: Our results demonstrate that SoCINS-OE plants have a repellent effect on aphids, and suggest an apparent link between 1,8-cineole, JA and trichome density. This study presents a viable and sustainable approach for aphid management by engineering the expression of 1,8-cineole synthase gene in plants, and underscores the potential usefulness of monoterpene synthase for pest control. © 2023 Society of Chemical Industry.

Identification and characterization of two general odorant binding protein genes from the oriental fruit moth, Grapholita molesta (busck).

Two novel general odorant-binding protein (GOBP) cDNAs (GmolGOBP1 and GmolGOBP2) were cloned and characterized from female antennal tissue of the oriental fruit moth, Grapholita molesta. We focused our investigation on this olfactory protein family by using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends-PCR (RACE-PCR). The full-length open reading frames of GmolGOBP1 and GmolGOBP2 were 492 and 483 nucleotides long, which encode 164 and 161 amino acid residue peptides, respectively. Protein signature analyses revealed that they each contained six conserved cysteines with an N-terminal signal sequence of 20 amino acids. The alignment of the two deduced protein sequences with other Lepidoptera GOBPs showed high sequence similarity (70-80%) with other full-length sequences from GenBank. Sequence similarity between the two GOBPs was only 48%, suggesting that the two proteins belong to different classes of lepidopteran GOBPs. RT-PCR analysis revealed that the two GOBP genes were expressed only in antennae of both sexes. Real-time PCR analysis further indicated that the transcript level of GmolGOBP1 was higher in males than in females, whereas the transcript level of GmolGOBP2 was higher in females than in males. Temporally, the two GOBP genes were expressed during the complete photoperiod (15L:9D). The highest transcript levels of GmolGOBP1 in both sexes and GmolGOBP2 in females were detected at the end of photophase and during scotophase. The expression of GmolGOBP2 in males remained at similar levels during the complete photoperiod. Based on these results, the possible physiological functions of GmolGOBPs are discussed.

Analysis of diastolic left ventricular wall shear stress in normal people of different age groups.

Background: Diastolic wall shear stress (WSS), assessed by using vector flow mapping (VFM), is the result of the interaction between the blood flow and the ventricular wall. This study aimed to evaluate the trend of left ventricular (LV) WSS in normal subjects. Methods and results: A total of 371 healthy volunteers were recruited and divided into four age groups (group I: 18-30 years; group II: 31-43 years; group III: 44-56 years; group IV: 57-70 years). LV WSS of different age groups was measured at each diastolic phase (P1: isovolumic diastolic period, P2: rapid filling period, P3: slow filling period, and P4:atrial contraction period) to evaluate the change trend of LV WSS. In each age group, LV WSS coincided with a trend of increasing-decreasing-increasing during P1-P4 (P < 0.05). Besides, among groups I, II, III, and IV, WSS of anterolateral, inferoseptal, and anteroseptal in P1 and WSS of inferolateral, inferoseptal, and anteroseptal in P4 all showed an increasing trend with age (P < 0.05). Regarding sex differences, women had greater diastolic WSS compared to men (P < 0.05). Conclusion: LV WSS showed a regular variation and had specific age- and sex-related patterns in different diastolic phases.

Peripheral Serum Exosomes Isolated from Patients with Acute Myocardial Infarction Promote Endothelial Cell Angiogenesis via the miR-126-3p/TSC1/mTORC1/HIF-1α Pathway.

Purpose: Angiogenesis is required for improving myocardial function and is a key factor in long-term prognosis after an acute myocardial infarction (AMI). Although exosomes are known to play a crucial role in angiogenesis, the role of peripheral exosomes in angiogenic signal transduction in patients with AMI remains unclear. Here, we explored the effect of exosomes extracted from the peripheral serum of AMI patients on angiogenesis and elucidated the downstream pathways. Patients and Methods: Serum exosomes were obtained from patients with AMI (AMI-Exo) and healthy individuals (Con-Exo). The exosomes were cocultured with human umbilical vein endothelial cells (HUVECs) in vitro, with aortic rings ex vivo, and were used to treat mouse hind-limb ischemia and mouse AMI model in vivo. Results: AMI-Exo raised HUVEC proliferation, tube formation, and migration, and enhanced microvessel sprouting from aortic rings compared to Con-Exo, both in vitro and ex vivo. Quantitative reverse transcription-polymerase chain reaction revealed that the abundance of miR-126-3p, a crucial regulator of angiogenesis, was increased in AMI-Exo. The inhibition of miR-126-3p decreased the benefits of AMI-Exo treatment, and miR-126-3p upregulation enhanced the benefits of Con-Exo treatment in HUVECs, aortic rings, the mouse hind-limb ischemia model, and the mouse AMI model. Knockdown and overexpression analyses revealed that miR-126-3p regulated angiogenesis in HUVECs by directly targeting tuberous sclerosis complex 1 (TSC1). Moreover, we found that miR-126-3p could inhibit TSC1 expression, which further activated mTORC1 signaling and increased HIF-1α and VEGFA expression, ultimately promoting angiogenesis. Conclusion: Collectively, our results provide a novel understanding of the function of exosomes in angiogenesis post AMI. We demonstrated that exosomes from the peripheral serum of AMI patients promote angiogenesis via the miR-126-3p/TSC1/mTORC1/HIF-1α signaling pathway.

Deep learning for the diagnosis of suspicious thyroid nodules based on multimodal ultrasound images.

Objectives: This study aimed to differentially diagnose thyroid nodules (TNs) of Thyroid Imaging Reporting and Data System (TI-RADS) 3-5 categories using a deep learning (DL) model based on multimodal ultrasound (US) images and explore its auxiliary role for radiologists with varying degrees of experience. Methods: Preoperative multimodal US images of 1,138 TNs of TI-RADS 3-5 categories were randomly divided into a training set (n = 728), a validation set (n = 182), and a test set (n = 228) in a 4:1:1.25 ratio. Grayscale US (GSU), color Doppler flow imaging (CDFI), strain elastography (SE), and region of interest mask (Mask) images were acquired in both transverse and longitudinal sections, all of which were confirmed by pathology. In this study, fivefold cross-validation was used to evaluate the performance of the proposed DL model. The diagnostic performance of the mature DL model and radiologists in the test set was compared, and whether DL could assist radiologists in improving diagnostic performance was verified. Specificity, sensitivity, accuracy, positive predictive value, negative predictive value, and area under the receiver operating characteristics curves (AUC) were obtained. Results: The AUCs of DL in the differentiation of TNs were 0.858 based on (GSU + SE), 0.909 based on (GSU + CDFI), 0.906 based on (GSU + CDFI + SE), and 0.881 based (GSU + Mask), which were superior to that of 0.825-based single GSU (p = 0.014, p< 0.001, p< 0.001, and p = 0.002, respectively). The highest AUC of 0.928 was achieved by DL based on (G + C + E + M)US, the highest specificity of 89.5% was achieved by (G + C + E)US, and the highest accuracy of 86.2% and sensitivity of 86.9% were achieved by DL based on (G + C + M)US. With DL assistance, the AUC of junior radiologists increased from 0.720 to 0.796 (p< 0.001), which was slightly higher than that of senior radiologists without DL assistance (0.796 vs. 0.794, p > 0.05). Senior radiologists with DL assistance exhibited higher accuracy and comparable AUC than that of DL based on GSU (83.4% vs. 78.9%, p = 0.041; 0.822 vs. 0.825, p = 0.512). However, the AUC of DL based on multimodal US images was significantly higher than that based on visual diagnosis by radiologists (p< 0.05). Conclusion: The DL models based on multimodal US images showed exceptional performance in the differential diagnosis of suspicious TNs, effectively increased the diagnostic efficacy of TN evaluations by junior radiologists, and provided an objective assessment for the clinical and surgical management phases that follow.