Tethered peptide activation mechanism of the adhesion GPCRs ADGRG2 and ADGRG4.

Adhesion G protein-coupled receptors (aGPCRs) constitute an evolutionarily ancient family of receptors that often undergo autoproteolysis to produce α and ß subunits1-3. A tethered agonism mediated by the 'Stachel sequence' of the ß subunit has been proposed to have central roles in aGPCR activation4-6. Here we present three cryo-electron microscopy structures of aGPCRs coupled to the Gs heterotrimer. Two of these aGPCRs are activated by tethered Stachel sequences-the ADGRG2-ß-Gs complex and the ADGRG4-ß-Gs complex (in which ß indicates the ß subunit of the aGPCR)-and the other is the full-length ADGRG2 in complex with the exogenous ADGRG2 Stachel-sequence-derived peptide agonist IP15 (ADGRG2(FL)-IP15-Gs). The Stachel sequences of both ADGRG2-ß and ADGRG4-ß assume a U shape and insert deeply into the seven-transmembrane bundles. Constituting the FXφφφXφ motif (in which φ represents a hydrophobic residue), five residues of ADGRG2-ß or ADGRG4-ß extend like fingers to mediate binding to the seven-transmembrane domain and activation of the receptor. The structure of the ADGRG2(FL)-IP15-Gs complex reveals the structural basis for the improved binding affinity of IP15 compared with VPM-p15 and indicates that rational design of peptidic agonists could be achieved by exploiting aGPCR-ß structures. By converting the 'finger residues' to acidic residues, we develop a method to generate peptidic antagonists towards several aGPCRs. Collectively, our study provides structural and biochemical insights into the tethered activation mechanism of aGPCRs.

Involvement of the oncogenic small nucleolar RNA SNORA24 in regulation of p53 stability in colorectal cancer.

Colorectal cancer (CRC) is a common malignant cancer worldwide. Although the molecular mechanism of CRC carcinogenesis has been studied extensively, the details remain unclear. Small nucleolar RNAs (snoRNAs) have recently been reported to have essential functions in carcinogenesis, although their roles in CRC pathogenesis are largely unknown. In this study, we found that the H/ACA snoRNA SNORA24 was upregulated in various cancers, including CRC. SNORA24 expression was significantly associated with age and history of colon polyps in CRC patient cohorts, with high expression associated with a decreased 5-year overall survival. Our results indicated that the oncogenic function of SNORA24 is mediated by promoting G1/S phase transformation, cell proliferation, colony formation, and growth of xenograft tumors. Furthermore, SNORA24 knockdown induced massive apoptosis. RNA-sequencing and gene ontology (GO) enrichment analyses were performed to explore its downstream targets. Finally, we confirmed that SNORA24 regulates p53 protein stability in a proteasomal degradation pathway. Our study clarifies the oncogenic role of SNORA24 in CRC and advance the current model of the role of the p53 pathway in this process.

The Effect of Impact Angle and Fall Height on Skull Fracture Patterns in Infants.

Skull fracture is a common finding for both accidental and abusive head trauma in infants and young children, and may provide important clues as to the energy and directionality of the event leading to the skull fracture. However, little is understood regarding the mechanics of skull fracture in the pediatric skull, and how accidental fall parameters contribute to skull fracture patterns. The objectives of this research were to utilize a newly developed linear elastic fracture mechanics finite element model of infant skull fracture to investigate the effect of impact angle and fall height on the predictions of skull fracture patterns in infants. Nine impact angles of right parietal bone impacts were simulated from three different heights onto a rigid plate. The average ± standard deviation of the distance between the impact location and fracture initiation site was 8.0 ± 5.9 mm. Impact angle significantly affected the fracture initiation site (p < 0.0001) and orientation (p < 0.0001). A 15 deg variation in impact angle changed the initiation site up to 47 mm. The orientation of the fracture pattern was dependent on the impact location and ran either horizontal or vertical toward the ossification center of the bone. Fall height significantly affected the fracture length (p = 0.0356). Specifically, at the same impact angle, a 0.3 m increase in fall height increased the skull fracture length by 21.39 ± 34.26 mm. These data indicate that environmental variability needs to be carefully considered when evaluating infant skull fracture patterns from low-height falls.

Long noncoding RNA lnc-RI regulates DNA damage repair and radiation sensitivity of CRC cells through NHEJ pathway.

A percentage of colorectal cancer (CRC) patients display low sensitivity to radiotherapy, which affects its therapeutic effect. Cancer cells DNA double-strand breaks (DSBs) repair capacity is crucial for radiosensitivity, but the roles of long noncoding RNAs (lncRNAs) in this process are largely uncharacterized. This study aims to explore whether lnc-RI regulates CRC cell growth and radiosensitivity by regulating the nonhomologous end-joining (NHEJ) repair pathway. CRC cells in which lnc-RI has been silenced showed lower cell growth and higher apoptosis rates due to increased DSBs and cell cycle arrest. We found that miR-4727-5p targets both lnc-RI and LIG4 mRNA and inhibit their expression. CRC cells showed increased radiosensitivity when lnc-RI was silenced. These results reveal novel roles for lnc-RI in both DNA damage repair and radiosensitivity regulation in CRC cells. Our study revealed that lnc-RI regulates LIG4 expression through lnc-RI/miR-4727-5p/LIG4 axis and regulates NHEJ repair efficiency to participate in DNA damage repair. The level of lnc-RI was negatively correlated with the radiosensitivity of CRC cells, indicates that lnc-RI may be a potential target for CRC therapy. We also present the first report of the function of miR-4727-5p.

CEP55, serving as a diagnostic marker gene for osteosarcoma, triggers the JAK2-STAT3-MMPs axis.

Background: Osteosarcoma (OS) stands as the prevailing form of primary bone cancer in clinical practice. Lack of effective treatment options and an overall poor prognosis are caused by the disease's exceptionally rare occurrence and unclear rationale. Objective: This study's goal is to determine diagnostic marker genes involved in the progression of OS and investigate related pathways and mechanisms with the purpose of offering effective methods for OS diagnostics and therapy. Methods: The Gene Expression Omnibus database provided the gene microarray data. Core genes were identified through differential expression analysis and WGCNA. Three techniques for machine learning, random forest, least absolute shrinkage and selection operator regression, and support vector machine recursive feature elimination, were used to further screen the core genes and obtain diagnostic marker genes for OS. The specificity and sensitivity of the diagnostic marker genes for OS diagnosis were evaluated using receiver operating characteristic curves. Western blotting analysis was used for preliminary validation of the diagnostic marker genes and their related pathways. Results: Two diagnostic marker genes were identified through screening, including CEP55 and VWF. Receiver operating characteristic curves have been utilized to assess the diagnostic and therapeutic effects of CEP55 and VWF on OS. Western blotting analysis preliminarily validated the overexpression of CEP55 in OS and its capacity to control MMP2 and MMP9 levels by activating the JAK2/STAT3 signaling pathway. Conclusion: At the first time, this research shows that CEP55 and VWF are more powerful diagnostic and predictive indicators for OS. CEP55 holds the capacity to activate the JAK2/STAT3 signaling pathway and modulate MMP2 and MMP9 levels, thereby positioning it as a promising target in OS treatment.

Screening of diagnostic biomarkers for ferroptosis-related osteoarthritis and construction of a risk-prognosis model.

Background: Osteoarthritis (OA) is the most prevalent and commonly chronic joint disease that frequently develops among the elderly population. It is not just a single tissue that is affected, but rather a pathology involving the entire joint. Among them, synovitis is a key pathological change in OA. Ferroptosis is a newly discovered form of cell death that results from the buildup of lipid peroxidation. However, the role and impact of it in OA are yet to be explored. Objective: The key to this work is to uncover the mechanisms of ferroptosis-related OA pathogenesis and develop more novel diagnostic biomarkers to facilitate the diagnostic and therapeutic of OA. Materials and methods: Download ferroptosis-related genes and OA synovial chip datasets separately from the FerrDB and Gene Expression Omnibus databases. Identify ferroptosis differentially expressed genes using R software, obtain the intersection genes through two machine learning algorithms, and obtain diagnostic biomarkers after logistic regression analysis. Verify the diagnostic and therapeutic efficacy of specific genes for OA through the construction of clinical risk prognostic models using ROC curves and nomogram. Simultaneously, correlations between specific genes and OA immune cell infiltration co-expression were constructed. Finally, verify the differential presentation of specific genes in OA and health control synovium. Results: Obtain 38 ferroptosis differentially expressed genes through screening. Based on machine learning algorithms and logistic regression analysis, select AGPS, BRD4, RBMS1, and EGR1 as diagnostic biomarker genes. The diagnostic and therapeutic efficacy of the four specific genes for OA has been validated by ROC curves and nomogram of clinical risk prognostic models. The analysis of immune cell infiltration and correlation suggests a close association between specific genes and OA immune cell infiltration. Further revealing the diagnostic value of specific genes for OA by the differential presentation analysis of their differential presentation in synovial tissue from OA and health control. Conclusion: This study identified four diagnostic biomarkers for OA that are associated with iron death. The establishment of a risk-prognostic model is conducive to the premature diagnosis of OA, evaluating functional recovery during rehabilitation, and guidance for subsequent treatment.

Mendelian randomization based on genome-wide association studies and expression quantitative trait loci, predicting gene targets for the complexity of osteoarthritis as well as the clinical prognosis of the condition.

Background: Osteoarthritis (OA) entails a prevalent chronic ailment, marked by the widespread involvement of entire joints. Prolonged low-grade synovial inflammation serves as the key instigator for a cascade of pathological alterations in the joints. Objective: The study seeks to explore potential therapeutic targets for OA and investigate the associated mechanistic pathways. Methods: Summary-level data for OA were downloaded from the genome-wide association studies (GWAS) database, expression quantitative trait loci (eQTL) data were acquired from the eQTLGen consortium, and synovial chip data for OA were obtained from the GEO database. Following the integration of data and subsequent Mendelian randomization analysis, differential analysis, and weighted gene co-expression network analysis (WGCNA) analysis, core genes that exhibit a significant causal relationship with OA traits were pinpointed. Subsequently, by employing three machine learning algorithms, additional identification of gene targets for the complexity of OA was achieved. Additionally, corresponding ROC curves and nomogram models were established for the assessment of clinical prognosis in patients. Finally, western blotting analysis and ELISA methodology were employed for the initial validation of marker genes and their linked pathways. Results: Twenty-two core genes with a significant causal relationship to OA traits were obtained. Through the application of distinct machine learning algorithms, MAT2A and RBM6 emerged as diagnostic marker genes. ROC curves and nomogram models were utilized for evaluating both the effectiveness of the two identified marker genes associated with OA in diagnosis. MAT2A governs the synthesis of SAM within synovial cells, thereby thwarting synovial fibrosis induced by the TGF-ß1-activated Smad3/4 signaling pathway. Conclusion: The first evidence that MAT2A and RBM6 serve as robust diagnostic for OA is presented in this study. MAT2A, through its involvement in regulating the synthesis of SAM, inhibits the activation of the TGF-ß1-induced Smad3/4 signaling pathway, thereby effectively averting the possibility of synovial fibrosis. Concurrently, the development of a prognostic risk model facilitates early OA diagnosis, functional recovery evaluation, and offers direction for further therapy.

Development and validation of a prognostic prediction model for cervical cancer patients treated with radical radiotherapy: a study based on TCGA database.

Background: Radiotherapy or concurrent chemoradiotherapy is the standard treatment for patients with locally advanced or inoperable cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC). However, treatment failure for CESC patients treated with radical radiotherapy still occurs due to local recurrence and distant metastasis. The previous prediction models were focused on all CESC patients, neglecting the prognostic differences under different treatment modalities. Therefore, there is a pressing demand to explore novel biomarkers for the prognosis and sensitivity of radiotherapy in CESC patients treated with radical radiotherapy. As a single biomarker has limited effect in stratifying these patients, our objective was to identify radioresponse-related mRNAs to ameliorate forecast of the prognosis for CESC patients treated with radical radiotherapy. Methods: Sample data on CESC patients treated with radical radiotherapy were obtained from The Cancer Genome Atlas (TCGA) database. We randomly separated these patients into a training and test cohorts using a 1:1 ratio. Differential expression analysis was carried out to identify radioresponse-related mRNA sets that were significantly dysregulated between complete response (CR) and radiographic progressive disease (RPD) groups, and univariate Cox regression analyses, least absolute shrinkage and selection operator (LASSO) method and multivariate Cox regression were performed to identify the radioresponse-related signature in the training cohort. we adopted survival analysis to measure the predictive value of the radioresponse-related signature both in the test and entire cohorts. Moreover, we developed a novel nomogram to predict the overall survival (OS) of CESC patients treated with radical radiotherapy. In addition, immune infiltration analysis and Gene Set Enrichment Analysis (GSEA) were conducted to preliminarily explore possible mechanisms. Results: This study included a total of 92 CESC patients subjected to radical radiotherapy. We developed and verified a risk score model based on radioresponse-related mRNA. The radioresponse-related mRNA signature and International Federation of Gynecology and Obstetrics (FIGO) stage were served as independent prognostic factors for CESC patients treated with radical radiotherapy. Moreover, a nomogram integrating radioresponse-related mRNA signature with FIGO stage was established to perform better for predicting 1-, 3-, and 5-year survival rates. Mechanically, the low-risk group under the risk score of this model had a better survival status, and the distribution of CD4 T cells was potentially involved in the regulation of radiotherapy response in CESC, leading to a better survival outcome in the low-risk group. Conclusions: This study presents a new radioresponse-related mRNA signature that shows promising clinical efficacy in predicting the prognosis of CESC patients treated with radical radiotherapy.

Research on doctor-patient communication teaching for oncology residents: a new teaching model.

BACKGROUND: At present, there is no specific teaching method for doctor-patient communication for oncology residents. This study combined BOPPPS (bridge-in, learning objective, pretest, participatory learning, posttest, and summary) teaching model and SBAR (situation-background-assessment-recommendation) communication model to try a new teaching and assessment model of doctor-patient communication, aiming to explore and improve the teaching method of doctor-patient communication for oncology residents. METHODS: Ninety residents were randomly divided into two groups: the experimental group (n = 45) was trained with the BOPPPS teaching model combined with the SBAR communication model, the routine teaching method was adopted in the control group (n = 45). Indicators of assessment included doctor-patient communication skills, satisfaction with teaching, and patient satisfaction. RESULTS: The scores for doctor-patient communication ability were significantly better in experimental group than control group (p < 0.05). The degree of satisfaction degree (very satisfied + satisfied) of the experimental group were higher than control group (p < 0.05). The overall teaching satisfaction of the experimental group was 100.00%, the control group was 77.78%, p < 0.001. Patients' satisfaction with the residents in the experimental group (93.3%) was significantly higher than control group (80.0%), p = 0.043. CONCLUSION: The application of the BOPPPS combined with the SBAR can effectively enhance residents' doctor-patient communication ability and improve teaching satisfaction and patient satisfaction. This new model can effectively improve resident physician‒patient communication ability, which is worth actively promoting in clinical teaching work.

The Effect of Unhealthy Food Packaging Information Boundaries on Consumer Purchasing Intentions.

Existing studies have examined unhealthy food packaging information, mainly focusing on aspects such as the content, color, and text, whilst paying less attention to the boundaries of information. This paper investigates unhealthy foods through three experiments, revealing that the presence (vs. absence) of packaging information boundaries on unhealthy foods has a negative impact on consumers' purchasing intentions (p = 0.040) (Experiment 1). The feeling of constraint mediates this effect (ß = -0.078, CI: [-0.1911, -0.0111]) (Experiment 2). Additionally, consumers with an independent self-construal exhibit reduced purchasing intentions when unhealthy food packaging information boundaries are present (vs. absent) (p < 0.001), whereas those with an interdependent self-construal show increased purchasing intentions under the same conditions (p = 0.024) (Experiment 3). This paper reveals the psychological mechanism and boundary conditions of unhealthy food packaging information boundaries affecting consumers' purchasing intention and provides practical inspiration for government policy-making related to unhealthy food packaging.

Molecular and circuit mechanisms underlying avoidance of rapid cooling stimuli in C. elegans.

The mechanisms by which animals respond to rapid changes in temperature are largely unknown. Here, we found that polymodal ASH sensory neurons mediate rapid cooling-evoked avoidance behavior within the physiological temperature range in C. elegans. ASH employs multiple parallel circuits that consist of stimulatory circuits (AIZ, RIA, AVA) and disinhibitory circuits (AIB, RIM) to respond to rapid cooling. In the stimulatory circuit, AIZ, which is activated by ASH, releases glutamate to act on both GLR-3 and GLR-6 receptors in RIA neurons to promote reversal, and ASH also directly or indirectly stimulates AVA to promote reversal. In the disinhibitory circuit, AIB is stimulated by ASH through the GLR-1 receptor, releasing glutamate to act on AVR-14 to suppress RIM activity. RIM, an inter/motor neuron, inhibits rapid cooling-evoked reversal, and the loop activities thus equally stimulate reversal. Our findings elucidate the molecular and circuit mechanisms underlying the acute temperature stimuli-evoked avoidance behavior.

Short-term effects of cupping and scraping therapy for chronic nonspecific low-back pain: A prospective, multicenter randomized trial.

BACKGROUND: As one of the most common musculoskeletal ailments, chronic nonspecific low-back pain (CNLBP) causes persistent disability and substantial medical expenses. Epidemiological evidence shows that the incidence rate of CNLBP in young and middle-aged people who are demanded rapidly recovery and social contribution is rising. Recent guidelines indicate a reduced role for medicines in the management of CNLBP. OBJECTIVE: The present study investigates the short-term effects of cupping and scraping therapy using a medicated balm, compared to nonsteroidal anti-inflammatory drug (NSAID) with a capsaicin plaster, in the treatment of CNLBP. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: We designed a prospective multicenter randomized clinical trial enrolling patients from January 1, 2022 to December 31, 2022. A total of 156 patients with CNLBP were randomized into two parallel groups. Diclofenac sodium-sustained release tablets were administered orally to participants in the control group for one week while a capsaicin plaster was applied externally. Patients in the test group were treated with cupping and scraping using a medical device and medicated balm. MAIN OUTCOME MEASURES: Primary outcome was pain recorded using the visual analogue scale (VAS). Two secondary outcomes were recorded using the Japanese Orthopedic Association low-back pain scale (JOA) and the traditional Chinese medicine (TCM) syndrome integral scale (TCMS) as assessment tools. RESULTS: Between baseline and postintervention, all changes in outcome metric scales were statistically significant (P < 0.001). Compared to the control group, patients in the test group had a significantly greater treatment effect in all outcome variables, as indicated by lower VAS and TCMS scores and higher JOA scores, after the one-week intervention period (P < 0.001). Further, according to the findings of multivariate linear regression analysis, the participants' pain (VAS score) was related to their marital status, age, smoking habits and body mass index. No adverse reactions were reported for any participants in this trial. CONCLUSION: The effectiveness of TCM combined with the new physiotherapy tool is superior to that of NSAID combined with topical plasters, regarding to pain intensity, TCM symptoms and quality of life. The TCM plus physiotherapy also showed more stable and long-lasting therapeutic effects. TRIAL REGISTRATION: This study was registered at Chinese Clinical Trial Registry (ChiCTR2200055655). Please cite this article as: He JY, Tu XY, Yin ZF, Mu H, Luo MJ, Chen XY, Cai WB, Zhao X, Peng C, Fang FF, Lü C, Li B. Short-term effects of cupping and scraping therapy for chronic nonspecific low-back pain: A prospective, multicenter randomized trial. J Integr Med. 2024; 22(1): 39-45.

Identification and Validation of a Necroptosis-Related Prognostic Signature for Kidney Renal Clear Cell Carcinoma.

Background: Necroptosis is progressively becoming an important focus of research because of its role in the pathogenesis of cancer and other inflammatory diseases. Our study is designed to anticipate the survival time of kidney renal clear cell carcinoma (KIRC) by constructing a prognostic signature of necroptosis-related genes. Materials: Clinical information and RNA-seq data were acquired from Renal Cell Cancer-European Union (RECA-EU) and The Cancer Genome Atlas- (TCGA-) KIRC, respectively. ConsensusClusterPlus was used to identify molecular subtypes, and the distribution of immune cell infiltration, anticancer drug sensitivity, and somatic gene mutations was studied in these subtypes. Subsequently, LASSO-Cox regression and univariate Cox regression were also carried out to construct a necroptosis-related signature. Cox regression, survival analysis, clinicopathological characteristic correlation analysis, nomogram, cancer stem cell analysis, and receiver operating characteristic (ROC) curve were some tools employed to study the prognostic power of the signature. Results: Based on the expression patterns of 66 survival-related necroptosis genes, we classified the KIRC into three subtypes (C1, C2, and C3) that are associated with necroptosis, which had significantly different tumor stem cell components. Among these, C2 patients had a longer survival time and enhanced immune status and were more sensitive to conventional chemotherapeutic drugs. Moreover, in order to predict the prognosis of KIRC patients, five genes (BMP8A, TLCD1, CLGN, GDF7, and RARB) were used to develop a necroptosis-related prognostic signature, which had an acceptable predictive potency. The results from Cox regression and stratified survival analysis revealed that the signature was an independent prognostic factor, whereas the nomogram and calibration curve demonstrated satisfactory survival time prediction based on the risk score. Conclusions: Three molecular subtypes and five necroptosis-related genes were discovered in KIRC using data from TCGA-KIRC and RECA-EU. Thus, a new biomarker and a potentially effective therapeutic approach for KIRC patients were provided in the current study.

SNORD15B and SNORA5C: Novel Diagnostic and Prognostic Biomarkers for Colorectal Cancer.

Colorectal cancer (CRC) is presenting a global public health problem with high incidence and mortality. Early diagnosis and treatment are the most important strategies to improve prognosis of this disease. Besides fecal occult blood test (FOBT) and colonoscopy, the most widely used methods for CRC screening currently, more effective methods for early diagnosis or prognostic prediction for CRC are needed. Small nucleolar RNAs (snoRNAs) is a class of noncoding RNAs (ncRNAs) playing crucial roles in carcinogenesis and considered to be promising tumor biomarker. In this study, we found that SNORD15B, SNORD48, and SNORA5C were significantly upregulated in CRC tissues. High levels of SNORD15B, SNORD48, or SNORA5C predicted poor clinical outcomes of CRC patients. Forced expression of SNORD15B or SNORA5C in CRC cells promoted proliferation and colony formation. In a further investigation, association between the level of SNORD15B/SNORA5C and clinicopathological parameters of CRC patient cohorts was analyzed based on data from The Cancer Genome Atlas (TCGA). We found that high expressions of SNORD15B and SNORA5C were significantly associated with age, lymphatic invasion, and history of colon polyps, and they were proved to be independent risk factors for survival of CRC patients. This study confirms that SNORD15B and SNORA5C have oncogenic effects in carcinogenesis of CRC. The findings suggest the two genes as potential diagnostic and prognostic biomarkers for CRC.

ALDH1B1 predicts poor survival for locally advanced nasopharyngeal carcinoma patients.

Background: Distant metastasis and local recurrence remain the major reasons of treatment failure in locally advanced nasopharyngeal carcinoma (NPC). Therefore, exploring novel biomarkers for prognosis and sensitivity of radiotherapy in locally advanced NPC is crucial. This retrospective study evaluates the expression and prognostic value of aldehyde dehydrogenase 1B1 (ALDH1B1) for locally advanced NPC patients. Methods: Sixty-seven locally advanced NPC patients and 22 chronic nasopharyngitis patients between September 2012 to November 2016 at The First Affiliated Hospital of University of South China were enrolled in this study. The expression of ALDH1B1 in tumor tissues were detected by using immunohistochemistry (IHC). Results: Significant difference was observed between NPC groups and Pharyngitis tissues groups, and NPC groups has a higher ratio of high ALDH1B1 expression. ALDH1B1 expression were significantly associated with age and radiotherapy response. The Kaplan-Meier analysis indicated that patients with high ALDH1B1 expression had a poor prognosis both in overall survival (OS) and progression-free survival (PFS). Univariate analysis found that age, radiotherapy response and ALDH1B1 expression were correlated with OS. Besides, factors affecting PFS are radiotherapy response and ALDH1B1 expression. Multivariate analysis revealed that radiotherapy response and ALDH1B1 expression were the independent prognostic factors for OS, whereas radiotherapy response was for PFS. Conclusions: The expression of ALDH1B1 was correlated with age and radiotherapy response. Patients with high ALDH1B1 expression show a poor prognosis both in OS and DFS. ALDH1B1 expression were the independent prognostic factors for OS.

Expression of Programmed Death Ligand-2 is associated with Prognosis in Nasopharyngeal Carcinoma Microenviroment.

Background: Although immune checkpoint inhibitors have opened a new mode of treatment for solid tumors, their efficacy in nasopharyngeal carcinoma (NPC) needs to be further investigated. Inhibitors of the PD-1/PD-L1 immune checkpoint are one of the hot topics in tumor immunotherapy. Programmed death ligand-2 (PD-L2) is a less studied ligand of PD-1 and has not yet been fully explored, especially in NPC. Understanding the clinical significance of PD-L2 expression, together with immune cell infiltration, might provide clues for biomarker screening in NPC immunotherapy. This study aimed to evaluate the role of PD-L2 as a prognostic factor for NPC patients as well as its role in immune regulation. Methods: Immunohistochemistry (IHC) was performed on a tissue microarray including 557 NPC specimens using PD-L2 antibody. The immune cell markers CD4, FOXP3 and CD68 were also stained and quantified. The expression of PD-L2 exhibited different spatial patterns among NPC tumor and stromal tissues. Results: A total of 90.8% of the cases showed membranous PD-L2 expression in tumors, and 80.8% showed membranous PD-L2 expression in stromal tissue. High stromal expression of PD-L2 predicted favorable overall and disease-free survival of NPC patients and was negatively correlated with tumor size, recurrence or metastasis and clinical stage. In contrast, high tumor abundance of PD-L2 correlated with poor disease-free survival, but had no obvious correlation with clinicopathological parameters. Multivariate analysis indicated that stromal PD-L2 was an independent and favorable prognostic factor. Furthermore, we found a positive correlation between stromal PD-L2 expression and the infiltration of CD68+ macrophages and CD4+Foxp3+ Treg cells in NPC stromal tissues (Pearson correlation=0.181 and 0.098, respectively). Conclusions: Our results suggest that different PD-L2 expression patterns have distinct predictive values. PD-L2 expressed on stromal cells might play a role in the regulation of NPC progression, and involve in immune activation in the tissue microenvironment and have an independent good prognosis for NPC patients.

MGSeg: Multiple Granularity-Based Real-Time Semantic Segmentation Network.

Recent works on semantic segmentation witness significant performance improvement by utilizing global contextual information. In this paper, an efficient multi-granularity based semantic segmentation network (MGSeg) is proposed for real-time semantic segmentation, by modeling the latent relevance between multi-scale geometric details and high-level semantics for fine granularity segmentation. In particular, a light-weight backbone ResNet-18 is first adopted to produce the hierarchical features. Hybrid Attention Feature Aggregation (HAFA) is designed to filter the noisy spatial details of features, acquire the scale-invariance representation, and alleviate the gradient vanishing problem of the early-stage feature learning. After aggregating the learned features, Fine Granularity Refinement (FGR) module is employed to explicitly model the relationship between the multi-level features and categories, generating proper weights for fusion. More importantly, to meet the real-time processing, a series of light-weight strategies and simplified structures are applied to accelerate the efficiency, including light-weight backbone, channel compression, narrow neck structure, and so on. Extensive experiments conducted on benchmark datasets Cityscapes and CamVid demonstrate that the proposed method achieves the state-of-the-art performance, 77.8%@50fps and 72.7%@127fps on Cityscapes and CamVid datasets, respectively, having the capability for real-time applications.

Kinetics model of DNA double-strand break repair in eukaryotes.

This manuscript outlines the kinetics of two main repair pathways of DNA double-strand break (DSB) in eukaryotes: non-homologous end joining (NHEJ) and homologous recombination repair (HRR). In this review, we discuss the precise study of recruitment kinetics of repair proteins based on the latest technologies in the past two decades. Then we simulate the theoretical description of the DNA repair process by mathematical models. In our study, the consecutive reactions chain (CRC) model and continuous-time random walk (CTRW) model have been unified by us, so that we can obtain the function of the number of intermediates with time in the same framework of equations, overcome the incompatibility between the two models. On this basis, we propose a data fitting workflow using these both models. Finally, we give an overview of different real-time quantitative methods and the new mechanism complexity that can be found from the corresponding dynamic models.

Cerebellar spreading depolarization mediates paroxysmal movement disorder.

Paroxysmal kinesigenic dyskinesia (PKD) is the most common paroxysmal dyskinesia, characterized by recurrent episodes of involuntary movements provoked by sudden changes in movement. Proline-rich transmembrane protein 2 (PRRT2) has been identified as the major causative gene for PKD. Here, we report that PRRT2 deficiency facilitates the induction of cerebellar spreading depolarization (SD) and inhibition of cerebellar SD prevents the occurrence of dyskinetic movements. Using Ca2+ imaging, we show that cerebellar SD depolarizes a large population of cerebellar granule cells and Purkinje cells in Prrt2-deficient mice. Electrophysiological recordings further reveal that cerebellar SD blocks Purkinje cell spiking and disturbs neuronal firing of the deep cerebellar nuclei (DCN). The resultant aberrant firing patterns in DCN are tightly, temporally coupled to dyskinetic episodes in Prrt2-deficient mice. Cumulatively, our findings uncover a pivotal role of cerebellar SD in paroxysmal dyskinesia, providing a potent target for treating PRRT2-related paroxysmal disorders.

Ligand recognition, unconventional activation, and G protein coupling of the prostaglandin E2 receptor EP2 subtype.

Selective modulation of the heterotrimeric G protein α S subunit-coupled prostaglandin E2 (PGE2) receptor EP2 subtype is a promising therapeutic strategy for osteoporosis, ocular hypertension, neurodegenerative diseases, and cardiovascular disorders. Here, we report the cryo-electron microscopy structure of the EP2-Gs complex with its endogenous agonist PGE2 and two synthesized agonists, taprenepag and evatanepag (CP-533536). These structures revealed distinct features of EP2 within the EP receptor family in terms of its unconventional receptor activation and G protein coupling mechanisms, including activation in the absence of a typical W6.48 "toggle switch" and coupling to Gs via helix 8. Moreover, inspection of the agonist-bound EP2 structures uncovered key motifs governing ligand selectivity. Our study provides important knowledge for agonist recognition and activation mechanisms of EP2 and will facilitate the rational design of drugs targeting the PGE2 signaling system.