The correctness and efficiency of the adapter connection are important factors affecting the quality and yield of the libraries in next-generation sequencing (NGS). Lengthening the sticky ends of the adapters and the targets can effectively improve the efficiency of ligation, however, it may cause a new trouble of adapter self-ligation. In this study, a strategy based on nickase is designed to limit or abolish long sticky-end adapters self-ligation. We firstly explored the enzymatic characteristics of Nt.BsmAI, a typical nickase. Then based on the working properties of Nt.BsmAI to design Y-adapters for NGS and evaluated the efficiency of this strategy for reducing long sticky-end adapter dimers. Moreover, the feasibility of Nb.BtsI (another nickase) was verified using the same strategy. This study provides new application for nickase in NGS library construction, and the possibility of using multi-base sticky end adapters in the future. NGS adapters with long sticky-ends paves a new avenue for enhancing the application performance of NGS, particularly for methylation sequencing, in biomedicine, with a higher efficiency in library preparation.

BACKGROUND:PD-1 and PD-L1 inhibitors have been shown to synergise with anti-angiogenic agents in non-small-cell lung cancer (NSCLC). We aimed to compare benmelstobart plus anlotinib with pembrolizumab in patients with previously untreated, driver gene-negative, PD-L1-positive, advanced NSCLC. METHODS:The blinded, randomised, controlled, phase 3 CAMPASS trial was conducted in 79 centres across China. Patients aged 18-75 years with stage IIIB-IV squamous or non-squamous NSCLC, no previous systemic treatment for advanced, recurrent or metastatic diseases, a PD-L1 tumour proportion score of 1% or greater, a life expectancy of 3 months or longer, at least one measurable lesion, and an Eastern Cooperative Oncology Group performance status of 0 or 1 were randomly assigned (2:1) to receive intravenous benmelstobart (1200 mg once on day 1) plus oral anlotinib (12 mg daily on days 1-14) or intravenous pembrolizumab (200 mg once on day 1) plus placebo every 3 weeks. Randomisation was done centrally and stratified by tumour histology, PD-L1 tumour proportion score, and brain metastases. Treatment allocation was open label for investigators and masked to patients and statisticians. The primary endpoint was progression-free survival as assessed by a blinded independent review committee per Response Evalutation Criteria in Solid Tumours version 1.1 in the intention-to-treat population (all randomly assigned patients). Safety was assessed in all randomly assigned patients who received at least dose of study drug. Results reported here are from a preplanned final analysis for progression-free survival. This ongoing study is closed to recruitment and is registered with ClinicalTrials.gov, NCT04964479. FINDINGS:Between Aug 6, 2021, and Dec 14, 2022, 531 patients were randomly assigned (354 to the benmelstobart plus anlotinib group and 177 to the pembrolizumab plus placebo group). 449 (85%) patients were male, 82 (15%) were female, and 493 (93%) were of Han ethnicity. Two patients in the benmelstobart plus anlotinib group and one patients in the pembrolizumab plus placebo group were untreated and therefore excluded from the safety population. After a median follow-up of 11·4 months (95% CI 9·4-13·1) for the benmelstobart plus anlotinib group and 10·6 months (9·0-13·0) for the pembrolizumab plus placebo group, median progression-free survival was 11·0 months (9·2-12·6) and 7·1 months (5·8-9·5), respectively (hazard ratio [HR] 0·70 [95% CI 0·54-0·90]; log-rank p=0·0057). Grade 3 or worse treatment-related adverse events occurred in 206 (59%) of 352 patients in the benmelstobart plus anlotinib group and 51 (29%) of 176 patients in the pembrolizumab plus placebo group, and the most frequent one was hypertension (90 [26%] vs five [3%]). Serious treatment-related adverse events occurred in 89 (25%) patients in the benmelstobart plus anlotinib group and 37 (21%) patients in the pembrolizumab plus placebo group, the most common of which were haemoptysis (nine [3%] vs none) and immune-mediated pulmonary diseases (eight [2%] vs five [3%]). Five (1%) treatment-related deaths occurred in the benmelstobart plus anlotinib group (two due to haemoptysis and one each due to immune-mediated pulmonary disease, disease progression, and infection pneumonia) and four (2%) occurred in the pembrolizumab plus placebo group (one each due to respiratory failure, pulmonary inflammation, disease progression, and myocardial injury). INTERPRETATION:Benmelstobart plus anlotinib showed longer progression-free survival than pembrolizumab plus placebo and no unexpected safety signals were reported, suggesting benmelstobart plus anlotinib as a potential first-line option in driver gene-negative, PD-L1-positive, advanced NSCLC. Longer term follow-up is needed to establish effects on overall survival. FUNDING:Chia Tai Tianqing Pharmaceutical Group.

[This corrects the article DOI: 10.3389/fmicb.2022.1046832.].

Importance:Patients with epidermal growth factor receptor (EGFR) gene variant nonsquamous non-small cell lung cancer (NSCLC) who have disease progression after prior EGFR tyrosine kinase inhibitor (TKI) therapy have limited treatment options, creating a need for more effective subsequent therapies. Objective:To provide final overall results of a trial assessing whether adding ivonescimab (a bispecific antibody targeting programmed cell death protein 1 and vascular endothelial growth factor) to chemotherapy improves overall survival in this population. Design, Setting, and Participants:Randomized, double-blind, placebo-controlled phase 3 trial conducted at 55 sites in China. From January 25 to November 2, 2022, a total of 322 adult patients with locally advanced or metastatic EGFR-variant nonsquamous NSCLC who had received prior EGFR-TKI therapy were enrolled. The data cutoff date was April 12, 2025. Interventions:Patients were randomized 1:1 to receive ivonescimab (20 mg/kg; n = 161) or placebo (n = 161) plus chemotherapy with pemetrexed and carboplatin once every 3 weeks for 4 cycles, followed by maintenance therapy. Main Outcomes and Measures:This final results report focuses on overall survival, the key secondary end point, tested in a hierarchical manner (the primary end point was progression-free survival assessed by an independent radiology review committee). Results:The 322 enrolled patients had a median age of 59.4 years, and 51.6% were female. During a median follow-up of 32.5 months, ivonescimab plus chemotherapy improved overall survival compared with chemotherapy alone (median survival, 16.8 months vs 14.1 months; stratified hazard ratio, 0.74; 95% CI, 0.58-0.95; P = .02). The absolute difference in median overall survival was 2.7 months. Estimated 30-month survival rates were 29.1% (95% CI, 22.1%-36.4%) with ivonescimab and 18.4% (95% CI, 12.8%-24.8%) with placebo. Grade 3 or higher treatment-emergent adverse events occurred in 67.1% and 54.7% of patients receiving ivonescimab and placebo, respectively. Conclusions and Relevance:Ivonescimab plus chemotherapy provided a statistically significant and clinically meaningful improvement in overall survival with an acceptable safety profile in patients with EGFR-variant NSCLC after EGFR-TKI therapy. Trial Registration:ClinicalTrials.gov Identifier: NCT05184712.

BACKGROUND:Gastric cancer (GC) remains a leading cause of cancer-related mortality worldwide. Solute carrier family 38 member 7 (SLC38A7), a lysosome-localized glutamine transporter, has been implicated in poor cancer prognosis and chemoresistance, but its biological role in GC is not well understood. This study aimed to investigate the biological function of SLC38A7 in GC progression and to elucidate the underlying signaling mechanism. METHODS:The expression pattern and clinical relevance of SLC38A7 were analyzed using public datasets from The Cancer Genome Atlas (TCGA) and the Genotype-Tissue Expression (GTEx) project, and validated in 60 paired GC tissues collected at the Chinese People's Liberation Army (PLA) General Hospital. Gain- and loss-of-function experiments were conducted in MKN-28 and MGC-803 to evaluate the effects of SLC38A7 on cell proliferation, migration, and invasion under glutamine-replete and glutamine-deprived conditions. Protein-protein interactions were assessed by co-immunoprecipitation and mass spectrometry. Site-directed mutagenesis was performed to identify key residues mediating SLC38A7 function. Downstream signaling was analyzed by western blotting and ubiquitination assays. A xenograft mouse model was established to determine the role of SLC38A7 in tumor growth in vivo. RESULTS:SLC38A7 expression was significantly upregulated in GC tissues and cell lines and was associated with poor patient prognosis. Functional assays demonstrated that SLC38A7 promoted GC cell proliferation, migration, and invasion, particularly under glutamine-deprived conditions. Mechanistically, SLC38A7 interacted with myosin heavy chain 9 (MYH9), facilitating glycogen synthase kinase-3β (GSK-3β) ubiquitination and degradation, which led to activation of the wingless/int-1 (Wnt)/β-catenin signaling pathway. Mutation of key SLC38A7 residues (E12 and Q3) disrupted its interaction with MYH9 and attenuated downstream signaling. Pharmacological inhibition of GSK-3β partially rescued the effects of SLC38A7 silencing in vitro. In vivo, SLC38A7 knockdown significantly suppressed tumor growth. CONCLUSION:SLC38A7 promotes gastric cancer progression by activating the Wnt/β-catenin signaling pathway through MYH9-dependent regulation of GSK-3β ubiquitination.

Background: Esophageal squamous cell carcinoma (ESCC) is a highly aggressive cancer with a poor prognosis, where immune evasion plays a central role in tumor progression and resistance to therapy. The underlying mechanisms of tumor-stroma interactions remain poorly understood, despite the relationship between epithelial-mesenchymal transition (EMT) and altered immune response having been suggested. This study aimed to investigate how phenotypic shifts in ESCC tumor cells contribute to immune modulation. Methods: We used multiplex immunofluorescence on 4-nitroquinoline 1-oxide (4-NQO)-induced multistage mouse ESCC models to characterize the local tumor microenvironment. Additionally, we integrated multiomics datasets, including spatial transcriptomics, single-cell RNA sequencing, and proteomics, from multistage human esophageal samples to investigate the underlying molecular mechanisms. These findings were further validated through in vitro cell line experiments and in vivo therapeutic models. Results: We identified an ESCC cell cluster with ectopic expression of CD11c (also known as integrin alpha X), in both mice and humans, probably formed via tumor protein p53 (TP53) inactivation, causing cancer cells to escape immune killing and gain malignant phenotypes. CD11c impaired cancer cell antigen presentation and fostered EMT through up-regulation of mothers against decapentaplegic homolog 3 (SMAD3) phosphorylation in human ESCC cell lines. Mechanistically, CD11c activated SMAD3 to suppress costimulatory factors CD80/CD86 and augmented immunosuppressive CD4+ T cell responses through aberrant major histocompatibility complex class II-mediated antigen presentation. Evaluation in humanized mouse models further confirmed that CD11c overexpression in ESCC resulted in immune evasion, tumor metastasis, and resistance to anti-programmed death ligand 1 (PD-L1) therapy, but could be rescued by combined treatment with anti-phospho-SMAD3. Conclusions: This study reveals a mechanism by which ectopic CD11c expression causes immunosuppression and contributes to the acquisition of malignant phenotypes in ESCC. Targeting the CD11c-SMAD3 axis may enhance the efficacy of existing immunotherapies, potentially improving the treatment outcomes of ESCC patients.

QuestionDoes adding ivonescimab to chemotherapy improve overall survival in patients with epidermal growth factor receptor (EGFR) gene variant non-small cell lung cancer after disease progression with EGFR tyrosine kinase inhibitor (TKI) therapy?FindingsIn this phase 3, randomized, double-blind trial, ivonescimab plus chemotherapy significantly improved overall survival (median, 16.8 vs 14.1 months) and the 30-month survival rate (29.1% vs 18.4%) compared with chemotherapy alone.MeaningThis regimen provides an effective post-EGFR-TKI treatment option, with a statistically significant survival benefit, a modest absolute improvement in median overall survival, and a more pronounced separation in long-term survival rate. ImportancePatients with epidermal growth factor receptor (EGFR) gene variant nonsquamous non-small cell lung cancer (NSCLC) who have disease progression after prior EGFR tyrosine kinase inhibitor (TKI) therapy have limited treatment options, creating a need for more effective subsequent therapies.ObjectiveTo provide final overall results of a trial assessing whether adding ivonescimab (a bispecific antibody targeting programmed cell death protein 1 and vascular endothelial growth factor) to chemotherapy improves overall survival in this population.Design, Setting, and ParticipantsRandomized, double-blind, placebo-controlled phase 3 trial conducted at 55 sites in China. From January 25 to November 2, 2022, a total of 322 adult patients with locally advanced or metastatic EGFR-variant nonsquamous NSCLC who had received prior EGFR-TKI therapy were enrolled. The data cutoff date was April 12, 2025.InterventionsPatients were randomized 1:1 to receive ivonescimab (20 mg/kg; n = 161) or placebo (n = 161) plus chemotherapy with pemetrexed and carboplatin once every 3 weeks for 4 cycles, followed by maintenance therapy.Main Outcomes and MeasuresThis final results report focuses on overall survival, the key secondary end point, tested in a hierarchical manner (the primary end point was progression-free survival assessed by an independent radiology review committee).ResultsThe 322 enrolled patients had a median age of 59.4 years, and 51.6% were female. During a median follow-up of 32.5 months, ivonescimab plus chemotherapy improved overall survival compared with chemotherapy alone (median survival, 16.8 months vs 14.1 months; stratified hazard ratio, 0.74; 95% CI, 0.58-0.95; P = .02). The absolute difference in median overall survival was 2.7 months. Estimated 30-month survival rates were 29.1% (95% CI, 22.1%-36.4%) with ivonescimab and 18.4% (95% CI, 12.8%-24.8%) with placebo. Grade 3 or higher treatment-emergent adverse events occurred in 67.1% and 54.7% of patients receiving ivonescimab and placebo, respectively.Conclusions and RelevanceIvonescimab plus chemotherapy provided a statistically significant and clinically meaningful improvement in overall survival with an acceptable safety profile in patients with EGFR-variant NSCLC after EGFR-TKI therapy.Trial RegistrationClinicalTrials.gov Identifier: NCT05184712 This randomized trial assesses the effect of ivonescimab added to chemotherapy on overall survival among patients with EGFR-variant non-small cell lung cancer (NSCLC) who had disease progression with epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) therapy.

The Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway plays a crucial role in innate immunity by inducing antiviral proteins in response to interferon signals. Marek's disease virus (MDV), a member of the alphaherpesvirus family, exerts potent tumorigenic and immunosuppressive effects. Recent studies have primarily focused on the tumorigenic mechanisms of MDV, and the mechanism of immune evasion has not been fully understood. In this study, we showed that MDV reduced the production of interferon-stimulated genes (ISGs) by inhibiting the phosphorylation and nuclear translocation of STAT1. Using a dual-luciferase reporter system, we screened for viral proteins that significantly suppress interferon-stimulated response element (ISRE) promoter activity. Meq overexpression markedly reduced ISRE promoter activity and ISG expression, whereas infection with Meq-deficient MDV induced higher ISG production in vitro and in vivo than infection with wild-type MDV. Meq also inhibited the phosphorylation and nuclear translocation of STAT1. Further experiments showed that Meq interacted with JAK1 and tyrosine kinase 2 (TYK2) and thereby inhibited JAK1-STAT1 interactions. Meq degraded TYK2 via a caspase-mediated pathway. The Meq-deficient MDV mutant replicated less efficiently than the wild-type MDV, both in vitro and in vivo. Collectively, these findings demonstrate that Meq played an immunosuppressive role in MDV by attenuating the JAK-STAT signaling pathway, which facilitated escape from innate immune surveillance mechanisms.

This study aimed to elucidate the global temporal and geographic characteristics of 29 cancers in older people aged ≥ 65 years, based on data from the Global Burden of Disease Study 2021. The average annual percentage changes (AAPCs) were calculated to estimate temporal trends of age-standardized incidence rates (ASIRs) and age-standardized mortality rates (ASMRs). Globally, there was an increase of 8.52 million cancer cases and 3.16 million cancer deaths among older people from 1990 to 2021. The ASIR of cancers combined presented an annually increased trend (AAPC: 0.49%), and regions with high sociodemographic index (SDI) experienced the highest increase (AAPC: 0.94%). Over the same period, the ASMR of cancers combined annually decreased (AAPC: -0.40%) globally, whereas regions with low SDI (AAPC: 0.32%) and low-middle SDI (AAPC: 0.48%) exhibited significantly increased ASMRs. Prostate cancer, lung cancer, and colorectal cancer were the three most common cancers for older people globally, and decreased relative inequalities were observed in higher-SDI countries from 1990 to 2021. For these three cancers, concentration index of ASMR respectively decreased from 0.26 to 0.06, from 0.20 to 0.17, and from 0.24 to 0.18. In contrast, the ASIR and ASMR of these cancers exhibited significant upward trends in lower-SDI regions. Our findings revealed that cancer burden for older people presented disparities globally, where higher-SDI countries faced a greater burden of cancer incidence and lower-SDI countries experienced an upward trend in cancer mortality. More attention should be given to prostate cancer, lung cancer, female breast cancer, and gastrointestinal cancers, especially in lower-SDI regions.

Feline coronavirus (FCoV) poses a significant threat to the lives of cats, and there is currently no commercial vaccines available. In the present study, a vaccine was developed using a human type 5 adenovirus vector to express the FCoV-I S protein (rAd5-FCoV-S) to induce the immunogenicity of rAd5-FCoV-S through oral and intramuscular immunization in mice and cats. Both vaccination methods stimulated a higher IgG antibody response. However, oral vaccination led to a significantly higher SIgA antibody level, which was 4.8 times and 2.4 times greater than that induced by intramuscular vaccination in mice and cats, respectively, with the highest level reaching 1:128. In addition, oral vaccination increased the count of IFN-γ-producing and IL-4-producing splenocytes in mice, effectively boosting cellular immune responses. Challenge protection experiments in cats showed that oral vaccination with rAd5-FCoV-S provided 100 % protection compared to a survival rate of only 33 % for unvaccinated cats. Compared to the PBS group, oral rAd5-FCoV-S administration substantially decreased the FCoV viral load within the feces, rectal tissues, and colon tissues of cats. Hematoxylin and eosin (HE) staining and immunohistochemical analysis of rectal and colonic tissues revealed that cats in the oral group exhibited minimal intestinal damage, whereas PBS cats presented significant inflammatory cell infiltration and shedding of intestinal epithelial cells. These findings demonstrate that oral administration of rAd5-FCoV-S induces a robust humoral immune response and a strong cell-mediated immune response in cats, thereby conferring immunity against FCoV infection.

BackgroundUntil now there has been no comprehensive data from extensive samples to evaluate the therapeutic potential of inetetamab in metastatic breast cancer (MBC) patients who had a history of trastuzumab treatment. Some previous studies had either small sample sizes from single-center, or partial enrolled patients without prior exposure to trastuzumab. This study aimed to provide a deep analysis of inetetamab-based therapy in this specific population.MethodsA multicenter retrospective study collected clinicopathological data from a total of 500 patients between Jul 2020 and Oct 2023. Progression-free survival (PFS) was estimated as the primary endpoint. Secondary endpoints included objective response rate (ORR), disease control rate (DCR) and adverse events (AEs). The association of risk factors with the effect of inetetamab treatment on PFS was evaluated using Cox proportional hazards regression analysis.ResultsIn the overall cohort, we observed a median PFS of 8.0 months, an ORR of 28.6% and a DCR of 89.2% (median treatment line: third line). Meanwhile, patients who received a combination treatment regime of inetetamab + tyrosine kinase inhibitors (TKIs) + chemotherapy achieved the most prolonged PFS of 9.0 months and the higher ORR of 29.3%. Ki67 index, distant lymph nodes metastasis and treatment strategy were independent predictors of PFS. The most common all-grade AEs were neutropenia (182/500, 36.4%) and leukopenia (157/500, 31.4%).ConclusionsInetetamab was promising in HER2-positive MBC patients who had prior exposure to trastuzumab, offering a new option for patients with a prior failure of trastuzumab.

BackgroundImmunotherapy plays a crucial role in the treatment of tumors. However, few studies have investigated the relationship between the expression of Programmed Cell Death Ligand 1 (PD-L1, CD274) in microvascular endothelial cells (MECs), including blood endothelial cells (BECs) and lymphatic endothelial cells (LECs), and immune cell infiltration within tissues.MethodsIn our study, we utilized data from The Cancer Genome Atlas, a mouse subcutaneous xenograft model, and immunofluorescence and immunohistochemical staining to investigate the relationship between PD-L1 expression in melanoma MECs at different tumor stages and the infiltration of CD8+ T cells in tumor and normal organs, under conditions with and without anlotinib treatment.ResultsWe found that PD-L1 expression was upregulated in tumor MECs, while anlotinib downregulated PD-L1 expression in both tumor and normal tissue MECs, corresponding with increased infiltration of CD8+ T cells in the tissues. Additionally, the antitumor effect of anlotinib was most pronounced when administered during the mid-stage of tumor development.ConclusionsThis study evaluated the most effective timing for anlotinib to downregulate PD-L1 expression in tumor and normal tissues to promote immune infiltration. Our findings may offer valuable insights for the clinical use of anlotinib and its potential side effects.

Vaccination is an effective strategy for preventing infectious diseases. Subunit vaccines offer more precise targeting and safer protection compared with traditional inactivated virus vaccines. However, due to their poor immunogenicity, subunit vaccines necessitate the use of adjuvants to stimulate the immune system. Adjuvants have long been incorporated into vaccines to enhance the body's immune response, allowing for reduced dosage and lower production costs. Despite the development of numerous vaccine adjuvants, few exhibit the necessary potency and low toxicity for clinical use, often due to limited efficacy or adverse side effects. This underscores the urgent need for novel human vaccine adjuvants that are safe, effective, and cost-efficient. Recent studies have identified certain natural polysaccharides as promising human vaccine adjuvants due to their immunostimulatory properties, low toxicity, and high safety profiles, which enhance both humoral and cellular immunity. These natural polysaccharides are primarily derived from traditional Chinese medicine (TCM) plants, bacteria, and yeast. This review comprehensively analyzes several promising polysaccharide adjuvants, discussing their clinical applications, market potential, and immunoregulatory activities. In summary, the future prospects of polysaccharides provide valuable insights for the application and development of vaccine adjuvants.

Immune checkpoint inhibitors (ICIs), principally represented by programmed death 1/programmed death ligand 1 (PD-1/PD-L1) inhibitors have been used in the first-line treatment of advanced lung cancer by Chinese Society of Clinical Oncology (CSCO) since 2018. Biomarkers like PD-L1 and tumor mutational burden (TMB) help to differentiate patients who benefited from ICI therapy. This study aimed to confirm and assess the predictive value of inflammatory markers in advanced lung cancer patients with the treatment of ICIs plus chemotherapy in the real world. The parameters of patients were collected in this retrospective study. Inflammatory markers were mainly calculated at different treatment periods. Hazard ratios (HR) and 95% confidence intervals (CI) were calculated with the univariate and multivariate Cox proportional hazard model. The Cox multivariate regression analysis indicated that a higher eosinophil-to-lymphocyte ratio (ELR) (HR 0.399, 95% CI: 0.189-0.842, P=0.02) and a higher monocyte-to-lymphocyte ratio (MLR) (HR 2.162, 95% CI: 1.116-4.191, P=0.02) at the time of maximizing therapeutic benefits, a higher lactate dehydrogenase (LDH) (HR 2.456, 95% CI: 1.334-4.523, P=0.004) at the time of disease progression were independent risk factors. A higher ELR and a lower MLR at maximizing therapeutic benefits and a lower LDH at disease progression were positive protectors for the progression-free survival (PFS) of advanced lung cancer treated with ICI combined with platinum-based chemotherapy.

Marek's disease virus (MDV) is an avian alphaherpesvirus associated with Marek's disease, an immunosuppressive and lymphoproliferative disease in chickens. The DNA sensing pathway mediates innate immune defense against infection by many DNA-containing pathogens, while viruses have evolved multiple strategies to evade the host immune response to survive in host cells. This study found that ectopic expression of MDV protein kinase US3 inhibited beta interferon (IFN-β) and interleukin-6 (IL-6) production induced by interferon-stimulatory and viral DNA. US3 was further shown to abolish the nuclear factor κB (NF-κB) activation. The US3 kinase activity was indispensable for its inhibitory function, as the kinase-dead US3 mutant (US3K220A) did not inhibit NF-κB activation. Further studies showed that US3 interacted with the Rel homology domains of the NF-κB subunits p65 and p50, which phosphorylated these transcription factors and blocked their nuclear translocation. Finally, US3 deficiency promoted IFN-β and IL-6 production, resulting in reduced viral replication and lower MDV-specific lesion incidence during MDV infection in chickens. Altogether, these findings reveal a novel mechanism for MDV to evade host antiviral immunity.IMPORTANCEMarek's disease virus (MDV) is an oncogenic avian alphaherpesvirus that causes an economically important disease affecting the health and welfare of poultry worldwide. Whereas human herpesviruses have been shown to evolve various strategies to inhibit the DNA sensing signaling for the evasion of the host's innate immunity, little is known regarding the mechanism for MDV to regulate this pathway. In this study, MDV US3 protein kinase was demonstrated to inhibit the activation of NF-κB in the DNA sensing pathway via binding to the Rel homology domains of the NF-κB subunits p65 and p50, which hyperphosphorylated these transcription factors and abolished their nuclear translocation. This is an important finding toward a better understanding of the functions of avian alphaherpesviruses encoded US3 protein kinase.

Sensitive and effective detection of epidermal growth factor receptor ( EGFR ) mutations is crucial for the early screening and diagnosis of non-small cell lung cancer (NSCLC). In this study, we assessed the sensitivity and specificity of the molecular switch technology combined with blocker primers for detecting EGFR exon 19 mutations. We demonstrated that this novel method allows real-time detection of mutated templates on a qPCR platform. Moreover, applying this method to cell-free DNA samples enhances the mutation detection rate.

LBA8502 Background: Anti-PD-(L) 1 monotherapy has been the standard first-line treatment for PD-L1 positive NSCLC, but its clinical benefit remains unsatisfactory. Benmelstobart (TQB2450) is a humanized monoclonal antibody against PD-L1 and anlotinib is a multikinase inhibitor that has been approved as the standard of care in the third-line treatment of NSCLC in China. This phase 3 study aimed to compare the efficacy of benmelstobart in combination with anlotinib and pembrolizumab as first-line treatment of PD-L1 positive aNSCLC. Methods: This is a randomized, single-blind, multicenter phase Ⅲ study (NCT04964479). Eligible patients (pts) were previous systemic treatment naïve, diagnosed with locally advanced or recurrent/metastatic NSCLC and had PD-L1 positive expression (defined as TPS ≥1%). Pts were randomized in a 2:1 ratio to receive either benmelstobart plus anlotinib (benmel+ anlo) or pembrolizumab plus placebo (pem+placebo). Anlotinib or placebo was administered orally at a dose of 12/0mg QD on days 1-14 of a 21-day cycle, while benmelstobart or pembrolizumab was given intravenously at a dose of 1200mg or 200mg on the first day of each cycle. The primary endpoint was progression-free survival (PFS) assessed by independent review committee (IRC). Results: Between August 2021 and December 2022, 531 pts were randomized (528 treated). At the data cutoff date of 20 May 2023, the median follow-up for PFS was 11.4 months for the benmel+anlo arm and 10.6 months for the pem+placebo arm. The study met its primary endpoint that the median PFS was significantly prolonged to 11.0 months (95% CI 9.2-12.6) in the benmel+anlo arm compared with 7.1 months (95% CI 5.8-9.5) in the pem+placebo arm (P = 0.007). The hazard ratio (HR) was 0.70 (95% CI 0.55-0.91). The HR for pts with squamous cell carcinoma and PD-L1 expression ≥50% was 0.63 (95% CI 0.46-0.86) and 0.60 (95% CI 0.41-0.88). The confirmed objective response rate was also obviously higher with combination therapy (57.3% vs. 39.6%; P < 0.001). The data for overall survival (OS) was immature. In total, 98.3% of pts in the benmel + anlo arm and 88.1% in the pem + placebo arm experienced at least one treatment-related adverse event (TRAE). The incidence of grade ≥3 TRAE was 58.5% and 29.0% in each group, respectively. Only 5.7%/3.7% of pts permanently discontinued benmelstobart/anlotinib since TRAE, while termination of pembrolizumab/placebo due to TRAE occurred in 8.0%/2.3% of pts. Conclusions: To our knowledge, this is the first phase III study to demonstrate the significant PFS benefit of a multikinase inhibitor plus an anti-PD-L1 mAb in the first-line treatment of PD-L1-positive aNSCLC compared to pembrolizumab. Tolerability is favourable with a lower incidence of treatment discontinuation due to TRAE. The data support this combination as a new option for these pts. Clinical trial information: NCT04964479 .