3'-epi-12ß-hydroxyfroside-mediated autophagy degradation of RIPK1/RIPK3 necrosomes leads to anergy of immunogenic cell death in triple-negative breast cancer cells.

Increasing studies have suggested that some cardiac glycosides, such as conventional digoxin (DIG) and digitoxin, can induce immunogenic cell death (ICD) in various tumors. We previously found that 3'-epi-12ß-hydroxyfroside (HyFS), a novel cardenolide compound isolated by our group, could induce cytoprotective autophagy through inactivation of the Akt/mTOR pathway. However, whether HyFS can induce ICD remains unknown. In this study, we extend our work to further investigate whether HyFS could induce both autophagy and ICD, and we investigated the relationship between autophagy and ICD in three TNBC cell lines. Unexpectedly, compared to DIG, we found that HyFS could induce complete autophagy flux but not ICD in three human triple-negative breast cancer (TNBC) cell lines and one murine TNBC model. Inhibition of HyFS-induced autophagy resulted in the production of ICD in TNBC MDA-MB-231, MDA-MB-436, and HCC38 cells. A further mechanism study showed that formation of RIPK1/RIPK3 necrosomes was necessary for ICD induction in DIG-treated TNBC cells, while HyFS treatment led to receptor-interacting serine-threonine kinase (RIPK)1/3 necrosome degradation via an autophagy process. Additionally, inhibition of HyFS-induced autophagy by the autophagy inhibitor chloroquine resulted in the reoccurrence of ICD and reversion of the tumor microenvironment, leading to more significant antitumor effects in immunocompetent mice than in immunodeficient mice. These findings indicate that HyFS-mediated autophagic degradation of RIPK1/RIPK3 necrosomes leads to inactivation of ICD in TNBC cells. Moreover, combined treatment with HyFS and an autophagy inhibitor may enhance the antitumor activities, suggesting an alternative therapeutic for TNBC treatment.

DIC/Oxyma-based accelerated synthesis and oxidative folding studies of centipede toxin RhTx.

Coupling reagents play crucial roles in the iterative construction of amide bonds for the synthesis of peptides and peptide-based derivatives. The novel DIC/Oxyma condensation system featured with the low risk of explosion displayed remarkable abilities to inhibit racemization, along with efficient coupling efficiency in both manual and automated syntheses. Nevertheless, an ideal reaction molar ratio in DIC/Oxyma condensation system and the moderate reaction temperature by manual synthesis remain to be further investigated. Herein, the synthetic efficiencies of different reaction ratios between DIC and Oxyma under moderate reaction temperature were systematically evaluated. The robustness and efficiency of DIC/Oxyma condensation system are validated by the rapid synthesis of linear centipede toxin RhTx. Different folding strategies were applied for the construction of disulfide bridges in RhTx, which was further confirmed in assays of circular dichroism and patch-clamp electrophysiology evaluation. This work establishes the DIC/Oxyma-based accelerated synthesis of peptides under moderate condensation conditions, which is especially useful for the manual synthesis of peptides. Besides, the strategy presented here provides robust technical supports for the large-scale synthesis and oxidative folding of RhTx.

The biological function of m6A reader YTHDF2 and its role in human disease.

N6-methyladenosine (m6A) modification is a dynamic and reversible post-transcriptional modification and the most prevalent internal RNA modification in eukaryotic cells. YT521-B homology domain family 2 (YTHDF2) is a member of m6A "readers" and its role in human diseases remains unclear. Accumulating evidence suggests that YTHDF2 is greatly implicated in many aspects of human cancers and non-cancers through various mechanisms. YTHDF2 takes a great part in multiple biological processes, such as migration, invasion, metastasis, proliferation, apoptosis, cell cycle, cell viability, cell adhesion, differentiation and inflammation, in both human cancers and non-cancers. Additionally, YTHDF2 influences various aspects of RNA metabolism, including mRNA decay and pre-ribosomal RNA (pre-rRNA) processing. Moreover, emerging researches indicate that YTHDF2 predicts the prognosis of different cancers. Herein, we focus on concluding YTHDF2-associated mechanisms and potential biological functions in kinds of cancers and non-cancers, and its prospects as a prognostic biomarker.

Current Understanding of the Involvement of the Insular Cortex in Neuropathic Pain: A Narrative Review.

Neuropathic pain is difficult to cure and is often accompanied by emotional and psychological changes. Exploring the mechanisms underlying neuropathic pain will help to identify a better treatment for this condition. The insular cortex is an important information integration center. Numerous imaging studies have documented increased activity of the insular cortex in the presence of neuropathic pain; however, the specific role of this region remains controversial. Early studies suggested that the insular lobe is mainly involved in the processing of the emotional motivation dimension of pain. However, increasing evidence suggests that the role of the insular cortex is more complex and may even be related to the neural plasticity, cognitive evaluation, and psychosocial aspects of neuropathic pain. These effects contribute not only to the development of neuropathic pain, but also to its comorbidity with neuropsychiatric diseases. In this review, we summarize the changes that occur in the insular cortex in the presence of neuropathic pain and analgesia, as well as the molecular mechanisms that may underlie these conditions. We also discuss potential sex-based differences in these processes. Further exploration of the involvement of the insular lobe will contribute to the development of new pharmacotherapy and psychotherapy treatments for neuropathic pain.

[Neurophysiological mechanisms and effects of emotional regulation on time perception].

Time is an important element for cognitive processes. Timing and time perception have been investigated by neuroscientists and psychologists for many years. It is well accepted that emotions could alter our experience of time. Previous studies of the emotional modulation on temporal perception focus primarily on behavioral and psychological experiments. In recent years, studies about the neurophysiological mechanisms of time perception have made some progress. Therefore, researchers started to explore how emotions influence our sense of time on the aspects of neural networks, neurotransmitters and synaptic plasticity. In this paper, we tried to review current studies about the effects of emotional regulation on time perception and the relevant neurophysiological mechanisms. This review will help us to deeply understand the neural mechanisms of time perception.

Differential modulation of electrical stimulation of periaqueductal gray and thalamus on nociceptive behaviors of rats.

Deep brain stimulation (DBS) is a surgical treatment which has shown remarkable therapeutic benefits for patients with a variety of neurologic conditions. As an important application, DBS has been used to treat intractable pain for over 60 years. Clinical studies have revealed that the selection of the stimulation sites depended on the types of pain. In this study, we selected ventrolateral periaqueductal gray (vlPAG) and ventral posterior lateral nucleus (VPL) as the target brain areas, which were widely used in clinical treatment of refractory pain, to clarify and compare the effects of vlPAG and VPL stimulation on different models of pain. Acute pain was evoked by thermal stimulation. The chronic inflammatory pain was produced by complete Freund's adjuvant (CFA) injection, while neuropathic pain was induced by spinal nerve ligation (SNL) surgery. Some important results emerged from this study: (1) in the experiment of normal rats, we found that unilateral vlPAG stimulation could lead to a significant increase of the thermal withdrawal threshold in bilateral hindpaws of rats, which means a significant bilateral analgesic action; (2) in the CFA test, both contralateral vlPAG and VPL stimulation significantly alleviated the thermal hyperalgesia, which exhibited analgesic effects to the chronic inflammatory pain; (3) in the SNL experiment, the results revealed that contralateral VPL stimulation could significantly abolish the mechanical allodynia induced by SNL, indicating remarkable analgesic effect to neuropathic pain. But the vlPAG stimulation did not have any effect on the mechanical allodynia. These results suggest that the electrical stimulation of the PAG works more effectively on nociceptive pain, including acute pain and chronic inflammatory pain. Besides, the VPL stimulation is much more sensitive for chronic pain, including chronic inflammatory pain and neuropathic pain.

Involvement of abscisic acid and salicylic acid in signal cascade regulating bacterial endophyte-induced volatile oil biosynthesis in plantlets of Atractylodes lancea.

The enormous biological diversity of endophytes, coupled with their potential to enhance the production of bioactive metabolites in plants, has driven research efforts focusing on endophytes. However, limited information is available on the impacts of bacterial endophytes on plant secondary metabolism and signaling pathways involved. This work showed that an endophytic Acinetobacter sp. ALEB16, capable of activating accumulation of plant volatile oils, also induced abscisic acid (ABA) and salicylic acid (SA) production in Atractylodes lancea. Pre-treatment of plantlets with biosynthetic inhibitors of ABA or SA blocked the bacterium-induced volatile production. ABA inhibitors suppressed not only the bacterium-induced volatile accumulation but also the induced ABA and SA generation; nevertheless, SA inhibitors did not significantly inhibit the induced ABA biosynthesis, implying that SA acted downstream of ABA production. These results were confirmed by observations that exogenous ABA and SA reversed the inhibition of bacterium-induced volatile accumulation by inhibitors. Transcriptional activities of genes in sesquiterpenoid biosynthesis also increased significantly with bacterium, ABA and SA treatments. Mevalonate pathway proved to be the main source of isopentenyldiphosphate for bacterium-induced sesquiterpenoids, as assessed in experiments using specific terpene biosynthesis inhibitors. These results suggest that Acinetobacter sp. acts as an endophytic elicitor to stimulate volatile biosynthesis of A. lancea via an ABA/SA-dependent pathway, thereby yielding additional insight into the interconnection between ABA and SA in biosynthesis-related signaling pathways.

Chronic administration of 5-HT1A receptor agonist relieves depression and depression-induced hypoalgesia.

Previous studies have shown that depressed patients as well as animal models of depression exhibit decreased sensitivity to evoked pain stimuli, and serotonin is indicated to be involved in depression-induced hypoalgesia. The purpose of this study was to investigate the potential role of 5-HT1A receptor in the depression-induced hypoalgesia. Acute or chronic administration of 5-HT1A receptor agonist, 8-OH-DPAT, was performed in olfactory bulbectomy (OB) and sham-operated rats. The depression-like behavior and pain thresholds were measured using open-field test and radiant heat thermal pain test, respectively. We found that acute administration of 8-OH-DPAT increased locomotor activity and pain thresholds in the sham rats but had no effect on the OB rats. In contrast, chronic administration of 8-OH-DPAT reduced locomotor activity and pain thresholds and restored them to normal level. Increased pain thresholds were also observed in the sham rats after the chronic administration. These results demonstrated that chronic administration of 8-OH-DPAT reversed the depression-induced decrease in pain sensitivity in rats, suggesting that 5-HT1A receptor may play a role in the depression-associated hypoalgesia.

Assessing cognitive biases induced by acute formalin or hotplate treatment: an animal study using affective bias test.

Pain, a universal and burdensome condition, influences numerous individuals worldwide. It encompasses sensory, emotional, and cognitive facets, with recent research placing a heightened emphasis on comprehending pain's impact on emotion and cognition. Cognitive bias, which encompasses attentional bias, interpretation bias, and memory bias, signifies the presence of cognitive distortions influenced by emotional factors. It has gained significant prominence in pain-related research. Human studies have shown that individuals experiencing pain exhibit cognitive bias. Similarly, animal studies have demonstrated cognitive bias in pain-induced states across various species and disease models. In this study, we aimed to investigate the memory bias displayed by rats experiencing acute pain, using the affective bias test (ABT) as a tool and administering either hotplate or formalin to induce acute pain. Our data showed that rats demonstrated a significant preference for the control treatment-related substrate over the substrate associated with formalin treatment (p < 0.001), an indication of the prominent memory bias stimulated by acute formalin injections. However, when exposed to substrates related to hotplate treatment and control treatment, the acute pain induced by the hotplate treatment failed to generate a statistically significant choice bias in rats (p = 0.674). Our study demonstrates that the negative emotions associated with acute pain can be reflected by memory bias in ABT, at least for formalin-induced acute pain. This finding will augment our comprehension of the emotional and cognitive aspects of acute pain.

Soreness Reminds Me of Grief: Patients With Chronic Pain Show Less Differentiated Representations of Emotional Feelings and Bodily States.

People experience similarities between emotional feelings and bodily states on a daily basis, but both the magnitude and pervasiveness of this experiential similarity vary across individuals. Inspired by previous findings that chronic pain (CP) is characterized by strengthened pain-affect coupling and reduced interoceptive accuracy, we conducted 2 cross-sectional studies to examine whether patients with CP would exhibit less differentiated perception and mental representation of emotional feelings and bodily states. In study 1 (N = 500), patients with CP and healthy controls (HCs) completed a self-report questionnaire that asked explicitly about the perceived similarity between 5 basic emotion categories and a series of bodily states. In study 2 (N = 73), a specially designed false memory test was administered to examine whether patients with CP would have reduced differentiation of concepts of negative emotion and somatic distress. We found that patients with CP perceived greater and more pervasive similarities between emotional feelings and bodily states, as indicated by higher questionnaire scores and denser, less specialized bipartite emotion-body networks, both associated with lower subjective interoceptive accuracy. Furthermore, patients with CP formed false memories of negative emotion words (eg, grief) more readily than HCs after memorizing somatic distress words (eg, soreness), as if they represented negative emotion and somatic distress as a single, enmeshed semantic category. Our findings extend previous literature by demonstrating reduced discrimination between emotional and bodily experiences in CP that is not restricted to pain-related emotional and sensory experiences and may be related to a fundamentally less differentiated interoception. PERSPECTIVES: This study shows that patients with chronic pain have a profoundly less differentiated perception and implicit conceptualization of emotional feelings and bodily states, which appears to be associated with altered interoception. These findings may provide new perspectives on why they often experience a stronger pain-affect coupling.

Differential relationships between autistic traits and anthropomorphic tendencies in adults and early adolescents.

Anthropomorphism, the attribution of human-like qualities (e.g., mental states) to nonhuman entities, is a universal but variable psychological experience. Adults with professionally diagnosed autism or high levels of subclinical autistic traits consistently show greater tendencies to anthropomorphize, which has been hypothesized to reflect 1) a compensatory mechanism for lack of social connectedness and 2) a persistence of childhood anthropomorphism into adulthood. Here, we directly tested these hypotheses in a general population sample consisting of both adults (N=685, 17-58 years old) and early adolescents (N=145, 12-14 years old) using the refined 9-item Anthropomorphism Questionnaire (AnthQ9), which measures both present and childhood anthropomorphic tendencies. We found that adults with heightened autistic traits reported increased present anthropomorphism (e.g., tend more to perceive computers as having minds), which held even after controlling for social connectedness. In contrast, adolescents with heightened autistic traits did not show increased present anthropomorphism, but rather reported reduced childhood anthropomorphism (e.g., less likely to perceive toys as having feelings) after controlling for social connectedness. We also found evidence that the present and childhood subscales of the AnthQ9 may tap into fundamentally different aspects of anthropomorphism. The results suggest that increased anthropomorphic tendencies in adults with heightened autistic traits cannot be explained solely by increased sociality motivation, but may be due to delayed development of anthropomorphism, although alternative possibilities of measurement problems cannot be ruled out. Implications for the measurement of anthropomorphism and its relation with theory of mind were also discussed.

Perioperative complication incidence and risk factors for retroperitoneal neuroblastoma in children: analysis of 571 patients.

BACKGROUND: Surgery plays an important role in the treatment of neuroblastoma. Perioperative complications may impact the course of neuroblastoma treatment. To date, comprehensive analyses of complications and risk factors have been lacking. METHODS: Patients with retroperitoneal neuroblastoma undergoing tumor resection were retrospectively analyzed between 2014 and 2021. The data collected included clinical characteristics, operative details, operative complications and postoperative outcomes. Risk factors for perioperative complications of retroperitoneal neuroblastoma were analyzed. RESULTS: A total of 571 patients were enrolled in this study. Perioperative complications were observed in 255 (44.7%) patients. Lymphatic leakage (28.4%), diarrhea (13.5%), and injury (vascular, nerve and organ; 7.5%) were the most frequent complications. There were three operation-related deaths (0.53%): massive hemorrhage (n = 1), biliary tract perforation (n = 1) and intestinal necrosis (n = 1). The presence of image-defined risk factors (IDRFs) [odds ratio (OR) = 2.09, P < 0.01], high stage of the International Neuroblastoma Risk Group staging system (INRGSS) (OR = 0.454, P = 0.04), retroperitoneal lymph node metastasis (OR = 2.433, P = 0.026), superior mesenteric artery encasement (OR = 3.346, P = 0.003), and inferior mesenteric artery encasement (OR = 2.218, P = 0.019) were identified as independent risk factors for perioperative complications. CONCLUSIONS: Despite the high incidence of perioperative complications, the associated mortality rate was quite low. Perioperative complications of retroperitoneal neuroblastoma were associated with IDRFs, INRGSS, retroperitoneal lymph node metastasis and vascular encasement. Patients with high-risk factors should receive more serious attention during surgery but should not discourage the determination to pursue total resection of neuroblastoma. Video Abstract (MP4 94289 KB).

The opioid placebo analgesia is mediated exclusively through µ-opioid receptor in rat.

Placebo analgesia is one of the most robust and best-studied placebo effects. Recent researches suggest that placebo analgesia activated the µ-opioid receptor signalling in the human brain. However, whether other opioid receptors are involved in the placebo analgesia remains unclear. We have previously evoked placebo responses in mice (Guo et al. 2010, 2011) and these mice may serve as a model for investigating placebo analgesia. In the present study, we tried to explore the site of action and types of opioid receptors involved in placebo response. Male Sprague-Dawley rats were trained with 10 mg/kg morphine for 4 d to establish the placebo analgesia model. This placebo analgesia can be blocked by injection of 5 mg/kg dose naloxone or by microinjection with naloxone (1, 3 or 10 µg/rat) into rostral anterior cingulate cortex (rACC). Then, animals were tested after intra-rACC microinjection of D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH(2) (CTOP, a selective µ-opioid receptor antagonist) or naltrindole (NTI, a highly selective δ-opioid receptor antagonist) or nor-binaltorphimine (nor-BNI, a highly selective κ-opioid receptor antagonist). Our results showed that CTOP, but not NTI or nor-BNI, could reduce the pain threshold in placebo analgesia rats. It may be concluded that rACC is the key brain region involved in placebo analgesia and the opioid placebo analgesia is mediated exclusively through µ-opioid receptor in rat.

[Multiple analysis of the difference in intestinal absorption between the main components and the extract of Glycyrrhiza uralensis].

The aim of this study is to investigate the rat intestinal absorption behavior of two main active components, liquiritin, glycyrrhizin and the extract of Glycyrrhiza uralensis. The rat intestinal perfusion model was employed. Concentrations of the compounds of the interest in the intestinal perfusate, bile and plasma samples were determined by HPLC and UPLC. At the same time, the intestinal enzymes incubation test and the partition coefficient determination, the absorption of liquiritin and glycyrrhizin alone and the extract were multiple analyzed. The results showed that the P(eff) (effective permeability) of liquiritin or glycyrrhizin alone or the extract was less than 0.3, which suggested their poor absorption in the intestine. The P(eff) of the two main active components or the extract was not significantly different in duodenum, jejunum, colon and ileum segment. The P(eff) of the glycyrrhizin in the extract had no significant difference in the four intestinal segments compared with the glycyrrhizin alone. The absorption of the liquiritin displayed significant difference (P < 0.05) at ileum segment compared with the liquiritin alone, while it had no markedly change in the other three segments. This phenomenon indicated that some ingredients in the extract might improve the absorption of liquiritin. Moreover, no parent compounds and their metabolites were found in the intestinal perfusate, bile and the plasma samples. The results demonstrated that the influence of the other ingredients in the extract on the two components might not increase the amount of liquiritin and glycyrrhizin in the bile and plasma within the duration of the test.

Application of Cognitive Bias Testing in Neuropsychiatric Disorders: A Mini-Review Based on Animal Studies.

Cognitive biases can arise from cognitive processing under affective states and reflect the impact of emotion on cognition. In animal studies, the existing methods for detecting animal emotional state are still relatively limited, and cognitive bias test has gradually become an important supplement. In recent years, its effectiveness in animal research related to neuropsychiatric disorders has been widely verified. Some studies have found that cognitive bias test is more sensitive than traditional test methods such as forced swimming test and sucrose preference test in detecting emotional state. Therefore, it has great potential to become an important tool to measure the influence of neuropsychiatric disorder-associated emotions on cognitive processing. Moreover, it also can be used in early drug screening to effectively assess the potential effects or side effects of drugs on affective state prior to clinical trials. In this mini-review, we summarize the application of cognitive bias tests in animal models of neuropsychiatric disorders such as depression, anxiety, bipolar disorder, and pain. We also discussed its critical value in the identification of neuropsychiatric disorders and the validation of therapeutic approaches.

Identification and expression profile of odorant-binding proteins in the parasitic wasp Microplitis pallidipes using PacBio long-read sequencing.

Microplitis pallidipes Szépligeti (Hymenoptera: Braconidae) is an important parasitic wasp of second and third-instar noctuid larvae such as the insect pests Spodoptera exigua, Spodoptera litura, and Spodoptera frugiperda. As in other insects, M. pallidipes has a chemosensory recognition system that is critical to foraging, mating, oviposition, and other behaviors. Odorant-binding proteins (OBPs) are important to the system, but those of M. pallidipes have not been determined. This study used PacBio long-read sequencing to identify 170,980 M. pallidipes unigenes and predicted 129,381 proteins. Following retrieval of possible OBP sequences, we removed those that were redundant or non-full-length and eventually cloned five OBP sequences: MpOBP2, MpOBP3, MpOBP8, MpOBP10, and MpPBP 429, 429, 459, 420, and 429 bp in size, respectively. Each M. pallidipes OBP had six conserved cysteine residues. Phylogenetic analysis revealed that the five OBPs were located at different branches of the phylogenetic tree. Additionally, tissue expression profiles indicated that MpOBP2 and MpPBP were mainly expressed in the antennae of male wasps, while MpOBP3, MpOBP8, and MpOBP10 were mainly expressed in the antennae of female wasps. MpOBP3 was also highly expressed in the legs of female wasps. Temporal profiles revealed that the expression of each M. pallidipes OBP peaked at different days after emergence to adulthood. In conclusion, we identified five novel odorant-binding proteins of M. pallidipes and demonstrated biologically relevant differences in expression patterns.

Title: Identification et profil d'expression des protéines de liaison aux odeurs chez la guêpe parasite Microplitis pallidipes à l'aide du séquençage à lecture longue PacBio. Abstract: Microplitis pallidipes Szépligeti (Hymenoptera : Braconidae) est une importante guêpe parasite des larves de noctuelles de deuxième et troisième stades telles que les insectes ravageurs Spodoptera exigua, Spodoptera litura et Spodoptera frugiperda. Comme d'autres insectes, M. pallidipes possède un système de reconnaissance chimiosensoriel, essentiel à la recherche de nourriture, à l'accouplement, à la ponte et à d'autres comportements. Les protéines de liaison aux odeurs (PLO) sont importantes pour le système, mais celles de M. pallidipes n'ont pas été déterminées. Cette étude a utilisé le séquençage à lecture longue PacBio pour identifier 170 980 unigènes de M. pallidipes et prédit 129 381 protéines. Après la récupération des séquences de PLO possibles, nous avons supprimé celles qui étaient redondantes ou pas de pleine longueur et avons finalement cloné cinq séquences de PLO, MpOBP2, MpOBP3, MpOBP8, MpOBP10 et MpPBP, respectivement de taille 429, 429, 459, 420 et 429 pb. Chaque PLO de M. pallidipes avait six résidus de cystéine conservés. L'analyse phylogénétique a révélé que les cinq PLO étaient situés à différentes branches de l'arbre phylogénétique. De plus, les profils d'expression tissulaire ont indiqué que MpOBP2 et MpPBP étaient principalement exprimés dans les antennes des guêpes mâles, tandis que MpOBP3, MpOBP8 et MpOBP10 étaient principalement exprimés dans les antennes des guêpes femelles. MpOBP3 était également fortement exprimé dans les pattes des guêpes femelles. Les profils temporels ont révélé que l'expression de chaque PLO de M. pallidipes culminait à différents jours après l'émergence à l'âge adulte. En conclusion, nous avons identifié cinq nouvelles protéines de liaison aux odeurs de M. pallidipes et démontré des différences biologiquement pertinentes dans les profils d'expression.

Immunotherapy combining tumor and endothelium cell lysis with immune enforcement by recombinant MIP-3α Newcastle disease virus in a vessel-targeting liposome enhances antitumor immunity.

BACKGROUND: Several agents for oncolytic immunotherapy have been approved for clinical use, but monotherapy is modest for most oncolytic agents. The combination of several therapeutic strategies through recombinant and nanotechnology to engineer multifunctional oncolytic viruses for oncolytic immunotherapy is a promising strategy. METHODS: An endothelium-targeting iRGD-liposome encapsulating a recombinant Newcastle disease virus (NDV), which expresses the dendritic cell (DC) chemokine MIP-3α (iNDV3α-LP), and three control liposomes were constructed. MIP-3α, HMGB1, IgG, and ATP were detected by western blotting or ELISA. The chemotaxis of DCs was examined by Transwell chambers. The phenotypes of the immune cells were analyzed by flow cytometry. The antitumor efficiency was investigated in B16 and 4T1 tumor-bearing mice. Immunofluorescence and immunohistochemistry were used to observe the localization of liposomes, molecular expression and angiogenesis. Synergistic index was calculated using the data of tumor volume, tumor angiogenesis and tumor-infiltrating lymphocytes. RESULTS: Compared with NDV-LP, treatment with iNDV3α-LP and NDV3α-LP induced stronger virus replication and cell lysis in B16 and 4T1 tumor cells and human umbilical vein endothelial cells (HUVECs) with the best response observed following iNDV3α-LP treatment. B16 and 4T1 cells treated with iNDV3α-LP produced more damage-associated molecular pattern molecules, including secreted HMGB1, ATP, and calreticulin. Moreover, iNDV3α-LP specifically bound to αvß3-expressing 4T1 cells and HUVECs and to tumor neovasculature. Tumor growth was significantly suppressed, and survival was longer in iNDV3α-LP-treated B16-bearing and 4T1-bearing mice. A mechanism study showed that iNDV3α-LP treatment initiated the strongest tumor-specific cellular and humoral immune response. Moreover, iNDV3α-LP treatment could significantly suppress tumor angiogenesis and reverse the tumor immune suppressive microenvironment in both B16-bearing and 4T1-bearing mice. CONCLUSIONS: In this study, iNDV3α-LP had several functions, such as tumor and vessel lysis, MIP-3α immunotherapy, and binding to αvß3-expressing tumor and its neovasculature. iNDV3α-LP treatment significantly suppressed tumor angiogenesis and reversed the tumor immunosuppressive microenvironment. These findings offer a strong rationale for further clinical investigation into a combination strategy for oncolytic immunotherapy, such as the formulation iNDV3α-LP in this study.

Ensemble encoding of nociceptive stimulus intensity in the rat medial and lateral pain systems.

BACKGROUND: The ability to encode noxious stimulus intensity is essential for the neural processing of pain perception. It is well accepted that the intensity information is transmitted within both sensory and affective pathways. However, it remains unclear what the encoding patterns are in the thalamocortical brain regions, and whether the dual pain systems share similar responsibility in intensity coding. RESULTS: Multichannel single-unit recordings were used to investigate the activity of individual neurons and neuronal ensembles in the rat brain following the application of noxious laser stimuli of increasing intensity to the hindpaw. Four brain regions were monitored, including two within the lateral sensory pain pathway, namely, the ventral posterior lateral thalamic nuclei and the primary somatosensory cortex, and two in the medial pathway, namely, the medial dorsal thalamic nuclei and the anterior cingulate cortex. Neuron number, firing rate, and ensemble spike count codings were examined in this study. Our results showed that the noxious laser stimulation evoked double-peak responses in all recorded brain regions. Significant correlations were found between the laser intensity and the number of responsive neurons, the firing rates, as well as the mass spike counts (MSCs). MSC coding was generally more efficient than the other two methods. Moreover, the coding capacities of neurons in the two pathways were comparable. CONCLUSION: This study demonstrated the collective contribution of medial and lateral pathway neurons to the noxious intensity coding. Additionally, we provide evidence that ensemble spike count may be the most reliable method for coding pain intensity in the brain.

The Effect of Electrical Stimulation-Induced Pain on Time Perception and Relationships to Pain-Related Emotional and Cognitive Factors: A Temporal Bisection Task and Questionnaire-Based Study.

Pain has not only sensory, but also emotional and cognitive, components. Some studies have explored the effect of pain on time perception, but the results remain controversial. Whether individual pain-related emotional and cognitive factors play roles in this process should also be explored. In this study, we investigated the effect of electrical stimulation-induced pain on interval timing using a temporal bisection task. During each task session, subjects received one of five types of stimulation randomly: no stimulus and 100 and 300 ms of non-painful and painful stimulation. Pain-related emotional and cognitive factors were measured using a series of questionnaires. The proportion of "long" judgments of a 1,200-ms visual stimulus duration was significantly smaller with 300 ms painful stimulation than with no stimulus (P < 0.0001) and 100 ms (P < 0.0001) and 300 ms (P = 0.021) non-painful stimulation. The point of subjective equality (PSE) did not differ among sessions, but the average Weber fraction (WF) was higher for painful sessions than for no-stimulus session (P = 0.022). The pain fear score correlated positively with the PSE under 100 ms non-painful (P = 0.031) and painful (P = 0.002) and 300 ms painful (P = 0.006) stimulation. Pain catastrophizing and pain anxiety scores correlated significantly with the WF under no stimulus (P = 0.005) and 100 ms non-painful stimulation (P = 0.027), respectively. These results suggest that electrical stimulation-induced pain affects temporal sensitivity, and that pain-related emotional and cognitive factors are associated with the processing of time perception.

Plants impact cellular immunity of caterpillars to an entomovirus.

BACKGROUND: Tri-trophic interactions among plants, insect herbivores and entomopathogens are one of the hot topics in ecology. Although plants have been shown to impact the interactions between herbivores and entomopathogens, it is still unclear how plants affect the cellular immunity of herbivores to entomopathogens. RESULTS: The number of hemocytes and the proportion of two main cell types (granular hemocytes and plasmatocytes), plasmatocyte-spreading rate, apoptosis rate, two Spodoptera exigua caspase (SeCasp-1, SeCasp-5) activities and gene expressions were all higher and the activities and gene expression of S. exigua inhibitor of apoptosis protein (SeIAP) were lower in nucleopolyhedrovirus (NPV)-infected caterpillars fed Ipomoea aquatica than those fed other plants or artificial diet. Scanning electron microscopy images were consistent with molecular patterns of immune responses. CONCLUSION: This study suggests that host plants affect the immune responses of herbivores to entomopathogens by manipulating the composition, morphology and apoptosis of herbivore hemocytes, which sheds light on the mechanisms that allow host plants to influence multi-trophic interactions. © 2021 Society of Chemical Industry.