The local cell-extrinsic and cell-intrinsic roles of endogenous interleukin-33 (IL-33) in tumor progression and metastasis have been controversial, which has delayed scrutiny of the systemic application of IL-33 in tumor immunoprevention and immunotherapy. A prominent concern regards its capacity in stabilizing the immunosuppressive phenotype of regulatory T (Treg) cells. Here, we report that systemic IL-33 treatment potently promotes the effects of prophylactic tumor vaccines and inhibits the progression of mouse solid tumors. Mechanistically, systemic IL-33 treatment reshapes the tumor immune microenvironment, including the increase in suppressor of tumorigenicity 2+ (ST2+) Treg cells. Unexpectedly, systemic IL-33 treatment constrains tumor growth in a Treg ST2-dependent manner, although the endogenous IL-33/ST2 axis in Treg cells promotes tumor growth. Indeed, IL-33 reprograms Treg cells toward a cytotoxic and inflammatory phenotype. Lastly, we fail to show that endogenous IL-33 promotes tumor growth. Thus, our findings elucidate a Treg ST2-dependent mechanism of the anti-tumor effect of systemic IL-33 application and support the use of IL-33 for cancer immunoprevention and immunotherapy.

Rabies virus (RABV) is a zoonotic pathogen with an almost 100% case-fatality rate and limited post-exposure prophylaxis options. mRNA-based antigen delivery represents a promising approach but requires further optimization. We prepared a cationic lipid nanoparticle (LNP) delivery system using the thin-film hydration method and characterized its physicochemical properties and delivery efficiency. An mRNA encoding a fusion antigen comprising rabies virus glycoprotein (G) and herpes simplex virus type 2 infected cell polypeptide 35 (ICP35), lacking canonical 5' and 3' untranslated region sequences, was constructed. In vitro protein expression of the G-ICP35 mRNA was confirmed. Immunogenicity was evaluated in BALB/c mice following three intramuscular immunizations with G-ICP35 mRNA, RABV G/N proteins, or their combination. Tail vein blood samples were collected on days 7, 21, and 39 after the final immunization, and rabies virus-specific antibody titers and T-cell responses were assessed. The cationic LNP formulation efficiently mediated intracellular delivery and expression of the G-ICP35 mRNA. In vivo, the mRNA-LNP formulation induced detectable humoral and cellular immune responses, including rabies virus-specific IgG production and antigen-specific T-cell activation. Notably, co-immunization with mRNA and subunit vaccines further enhanced immune responses compared with either vaccine alone, achieving peak rabies virus-specific IgG titers of 694.5 IU/mL. These findings demonstrate the feasibility of a DCChol-based mRNA-LNP formulation for fusion antigen delivery and support further investigation of combined mRNA and subunit vaccine strategies.

Herpes simplex virus type 1 (HSV-1) triggers significant inflammation and immune dysregulation, particularly in immunocompromised hosts. MicroRNA-221 (miR-221) is implicated in viral pathogenesis and inflammatory responses, yet its role in HSV-1 infection remains undefined. This study demonstrates that miR-221 suppresses HSV-1 replication, with 100 nM miR-221 reducing plaque formation by approximately 70

Herpes simplex virus type 1 (HSV-1) is a very concerning pathogen due to its ability to persist in the host’s nervous system and continuously interfere with the immune system, which complicates treatment. Therefore, the development of an effective HSV-1 vaccine is crucial. In this study, we focused on an HSV-1 mutant strain, M6, which includes several deleted genes associated with viral infection virulence and latent infection function, and explored its infection of macrophages and immunological characteristics. The study found that both the attenuated strain M6 and the wild-type strain infect macrophages through the binding of the gD protein to the HVEM receptor on the macrophage surface. Compared to the wild-type strain, the attenuated M6 strain induced a milder immune response, characterized by the lower expression of immune signaling molecules and inflammatory cytokine levels. Upon reintroducing macrophages infected with the two strains into mice, the M6 strain induced lower levels of inflammatory cytokines and higher levels of chemokines in spleen cells and also slightly lower humoral and cellular immune responses than the wild-type strain. Further histopathological analysis revealed that mice in the attenuated M6 group showed more stable body weight changes and milder pathological damage in immune organs such as the liver, spleen, and lymph nodes. In conclusion, the attenuated M6 strain exhibits good immunogenicity and mild pathological side effects, suggesting its potential as an effective immunogen.

BACKGROUND:Genes associated efferocytosis play a critical role in tumor invasion, metastasis, and clinical outcomes in several malignancies. However, their prognostic relevance in cervical squamous carcinoma (CESC) is currently unclear. METHODS:Data from the Cancer Genome Atlas (TCGA) database were used to create the TCGA-CESC cohort. Additional CESC-related datasets (GSE44001 and GSE168652) were retrieved from the Gene Expression Omnibus database. Single-cell analysis of the GSE168652 dataset was conducted to identify differentially expressed genes (DEGs), associated with efferocytosis (ER-DEGs). CESC samples from the TCGA cohort were classified into clusters, and prognosis-related DEGs (PR-DEGs) were identified by comparing these clusters. The intersection of ER-DEGs and PR-DEGs generated a subset of prognosis-efferocytosis related DEGs (PER-DEGs). Subsequently, a risk model was developed using univariate Cox regression and least absolute shrinkage and selection operator analysis. Independent prognostic factors were further analyzed using Cox regression analysis. Functional pathways were examined through gene set enrichment analysis. RESULTS:A total of 15 cell clusters were identified from the GSE168652 dataset and categorized into seven distinct cell types. Comparative analysis indicated 774 ER-DEGs between groups with high and low ERG expression levels. From these, 332 PER-DEGs were identified. A prognostic risk model was constructed based on seven characteristic genes: ITGA5, SNRPF, EGLN1, NDUFB7, NDUFA2, SRGAP3, and PCNA. Risk score and pathologic-N emerged as independent prognostic factors. These genes were found to be associated with multiple pathways implicated in the pathogenesis of CESC. CONCLUSION:Seven efferocytosis-related prognostic genes were identified as prognostic markers for CESC, providing a scientific basis for further research on their role in disease progression.

OBJECTIVE:To present reproductive and oncological outcomes of radical trachelectomy (RT) in patients with cervical adenocarcinomas (AC). METHODS:This retrospective study included 51 patients with cervical AC who underwent RT at Peking Union Medical Hospital from January 1, 2005 to June 1, 2023. RESULTS:Five patients (9.8%) experienced cervical stenosis following RT, which likely occurred in cases of abdominal RT (50%) and virginal prophylactic cerclage (33.33%) and those without copper T intrauterine devices during RT (20%). In total, 30 patients (58.82%) attempted to conceive, and 11 (36.67%) succeeded. Five patients (45.45%) achieved pregnancy with fertility assistance. The mean surgery-pregnancy interval was 27 months (range, 17-118). Two preterm and two full-term births were achieved. With a median follow-up of 50 months (range, 7-238), seven patients (13.73%) experienced recurrence and three (5.88%) died. Six of seven patients relapsed beyond the residual cervix. The cancer recurrence rate (CRR) was 5.88% for patients with pre-cervical conization and 17.65% for those with biopsy (p = 0.250); 11.63% had human papillomavirus-associated (HPVA) disease and 25% had non-HPVA (NHPVA) (p = 0.313). The cancer death rate (CDR) was 4.65% with HPVA and 12.50% with NHPVA (p = 0.386); 13.63% had the endogenous type and 0 had the exogenous type (p = 0.04). Chemotherapy in patients with risk factors resulted in better CRR and CDR than in those without (5.88% vs. 17.65%, 0% vs. 8.82%). The cumulative 5-year recurrence-free survival (RFS) and overall survival rates were 82.03% and 94.39%, respectively. CONCLUSION:RT in patients with AC led to an acceptable pregnancy rate but a higher CRR and lower 5-year RFS. Careful patient selection for RT, combined with adjuvant chemotherapy when indicated, is crucial to optimize the balance between reproductive and oncological outcomes in AC.

ABSTRACT To effectively control viral pandemics, a multifaceted approach incorporating diverse vaccination strategies may be imperative. Our study demonstrates that both mucosal and non-mucosal immunization of mice with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) subunit vaccine enriched with thymic stromal lymphopoietin (TSLP) results in potent vaccine-specific systemic IgG and IgG1 antibody responses, but only mucosal immunization can induce robust mucosal IgA antibody and neutralizing antibody against SARS-CoV-2 wild-type, Alpha B.1.1.7, and Delta B.1.617.2 variant strains. We found that the immune-enhancing effect of TSLP relies on promoting dendritic cell (DC) migration and activation, which in turn enhances the germinal center (GC) responses. Furthermore, intranasal administration of the TSLP-adjuvanted vaccine protected against challenges from both SARS-CoV-2 wild-type and Delta B.1.617.2 strains. Our findings offer valuable insights into the importance of TSLP as a critical mucosal adjuvant in enhancing mucosal adaptive immunity by modulating DC-GC crosstalk, hence enhancing vaccination effectiveness. IMPORTANCE The current development and implementation of subunit vaccines in clinical practice encounter a number of challenges. The most significant difficulties are the scarcity of effective adjuvants and the fact that most vaccination programs rely on non-mucosal injections, which frequently fail to establish mucosal adaptive immunity and hence restrict the potential to protect against respiratory viral infections. Our research indicates that TSLP can effectively boost SARS-CoV-2 subunit vaccine-specific systemic and mucosal humoral immunity in mice after intranasal immunization, protecting them from SARS-CoV-2 wild-type and Delta variant infection. The mucosal adjuvant effects of TSLP depend on DC migration and activation, which boosts germinal center responses. Therefore, supplementation of mucosal subunit vaccines with TSLP should be considered in vaccine development, particularly when the vaccine is administered to children and the elderly. Our findings provide new evidence for the development of mucosal adjuvants for infectious diseases, potentially facilitating future vaccine development.

Live attenuated and vero-cell-inactivated Japanese Encephalitis vaccines (LJEV, IJEV) have been in common use in young children in China since 1989 and 2004, associated with large reductions in Japanese encephalitis (JE) incidence. In 2013, northern China reported JE outbreaks among adults born before JE vaccine availability, a trend that worsened in 2017-2018. We conducted an open-label, randomized, controlled trial (ChiCTR2500103235) to assess the immunogenicity, immune persistence, and safety of three JE vaccine schedules in 40-69-year-olds to provide evidence for adult targeted JE immunization efforts. Outcomes were seroconversion proportions and seropositive prevalences; adverse events were monitored. The vaccines were immunogenic with no significant difference between vaccination groups. Seropositivity remained above 80% at one year post-vaccination. No serious adverse events occurred. All three schedules had good, persistent immunogenicity and favorable safety profiles in 40-69-year-old adults, providing evidence supporting vaccinating adults in response to the emergence of adult JE in northern China.

In this study, we aimed to assess the efficacy of polymerase chain reaction (PCR) targeting Leishmania kinetoplast minicircle DNA (kDNA) for diagnosing visceral leishmaniasis-associated hemophagocytic lymphohistiocytosis (VL-HLH). Three pediatric patients diagnosed with VL-HLH via the microscopic examination of bone marrow smears were selected to explore the diagnostic value of kDNA PCR in Leishmania species identification and parasite load quantification in three cases of visceral leishmaniasis. Total DNA was extracted from the bone marrow samples, and PCR testing targeting the Leishmania kDNA was conducted. The amplified PCR products were sequenced and analyzed by using National Center for Biotechnology Information Basic Local Alignment Search Tool for Leishmania species identification and homology cluster analysis. Furthermore, universal primers and probes were designed to quantify the kDNA load in patient samples through real-time quantitative PCR (qPCR) testing. Polymerase chain reaction testing targeting the kDNA gene yielded a specific 139 base pairs band in all samples. Real-time qPCR testing targeting the kDNA gene revealed significant amplification in all patients. The kDNA copy numbers for the three patients were 3,830/µL, 150/µL, and 77,168/µL, respectively. Homology analysis revealed 97.8% similarity between Patients 1 and 2, as well as 87.1% and 87.8% similarity between Patients 1 and 3 and Patients 2 and 3, respectively. All sequences were identified as belonging to the genus Leishmania, with Patients 1 and 2 clustering with Leishmania donovani sequences and Patient 3 clustering with Leishmania infantum. Polymerase chain reaction testing targeting the kDNA gene is a rapid, accurate, and sensitive method for detecting and quantifying Leishmania in VL-HLH patients. It also facilitates species identification through sequencing, demonstrating its potential as a valuable tool in clinical diagnostics.

Men who have sex with men (MSM) face a heightened risk of HIV, presenting a significant public health issue. This meta-analysis evaluates the effectiveness of HIV health education for MSM by examining changes in knowledge, attitudes, and behaviors across different cultures and regions. The study also explores heterogeneity sources through subgroup and sensitivity analyses to guide future research. Searches were conducted in databases including China Knowledge, Wipo, Wanfang, PubMed, and Web of Science, covering studies up to December 1, 2023. Included were controlled pre-post trials on HIV education's impact on MSM. Independent data extraction and analysis were completed by two researchers using RevMan 5.7. Pre-registration was done with PROSPERO. Twenty-seven studies met criteria. Post-intervention, significant improvements were noted in HIV knowledge (OR: 0.49; 95% CI: 0.44-0.56), condom use over six months (OR: 0.47; 95% CI: 0.38-0.58), and condom use in recent same-sex encounters (OR: 0.44; 95% CI: 0.29-0.66). Larger samples (>500) or long-term interventions (>1 year) enhanced homogeneity (I2 = 0%). In conclusion, these interventions significantly boost knowledge and condom use, identifying sample size and intervention duration as primary heterogeneity influences, guiding future strategy optimization.

Ataxia-telangiectasia (A-T) is a hereditary neurodegenerative disorder caused by mutations in the ATM (ataxia-telangiectasia mutated) gene. Although existing rodent models reproduce some of the multi-systemic features of A-T, they notably fail to recapitulate the severe neurological manifestations, particularly the profound cerebellar atrophy and associated ataxia. To address this limitation, we have generated ATM-deficient rhesus macaques using CRISPR-Cas9. These macaques exhibit hallmark features of A-T, including growth retardation, lymphopenia, elevated a-fetoprotein levels, oculocutaneous telangiectasias, heightened sensitivity to ionizing radiation, and most critically, cerebellar atrophy, Purkinje cell loss, and early-stage cerebellar neurodegeneration leading to significant motor impairments. Single-nucleus transcriptomic profiling of the cerebellum revealed pronounced gene expression changes associated with ATM deficiency, particularly in molecular layer interneurons (MLIs), which are implicated in Purkinje cell loss. This non-human primate model provides deeper insights into the pathogenesis of A-T and represents a promising and valuable platform for developing therapeutic strategies.

Novel coronavirus pneumonia, also known as coronavirus disease 2019 (COVID-19), is caused by sub-severe acute respiratory syndrome type 2 coronavirus (SARS-CoV-2) infection. The spike (S) protein of SARS-CoV-2 binds to angiotensin-converting enzyme 2 (ACE2) receptors widely expressed on the surface of human cells leading to life-threatening respiratory infections. A serious hazard to human health is posed by the lack of particular treatment medications for this virus infection. We advocate the creation of high-affinity antibodies using the receptor binding domain (RBD) of S protein as a specific antigenic epitope to develop a drug that can precisely target therapy COVID-19 because SARS-CoV-2 infection of the host cells is dependent on S protein binding to ACE2. Finally, we obtained high-affinity antibodies 14F4HL and 14E3HL that have high affinity with RBD and well-drug-forming properties, suitable for further humanization studies. Thus, monoclonal antibodies that neutralize the S protein were identified in our study, which may provide new insights for the development of COVID-19 therapeutic drugs.

Recent studies have indicated that sequentially administering SARS-CoV-2 vaccines can result in increased antibody and cellular immune responses. In this study, we compared homologous and heterologous immunization strategies following two doses of inactivated vaccines in a mouse model. Our research demonstrates that heterologous sequential immunization resulted in more immune responses displayed in the lymph node germinal center, which induced a greater number of antibody-secreting cells (ASCs), resulting in enhanced humoral and cellular immune responses and increased cross-protection against five variant strains. In further single B-cell analysis, the above findings were supported by the presence of unique B-cell receptor (BCR) repertoires and diversity in CDR3 sequence profiles elicited by a heterologous booster immunization strategy.

The circulating severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) variant presents an ongoing challenge for surveillance and detection. It is important to establish an assay for SARS-CoV-2 antibodies in vaccinated individuals. Numerous studies have demonstrated that binding antibodies (such as S-IgG and N-IgG) and neutralizing antibodies (Nabs) can be detected in vaccinated individuals. However, it is still unclear how to evaluate the consistency and correlation between binding antibodies and Nabs induced by inactivated SARS-CoV-2 vaccines. In this study, serum samples from humans, rhesus macaques, and hamsters immunized with inactivated SARS-CoV-2 vaccines were analyzed for S-IgG, N-IgG, and Nabs. The results showed that the titer and seroconversion rate of S-IgG were significantly higher than those of N-IgG. The correlation between S-IgG and Nabs was higher compared to that of N-IgG. Based on this analysis, we further investigated the titer thresholds of S-IgG and N-IgG in predicting the seroconversion of Nabs. According to the threshold, we can quickly determine the positive and negative effects of the SARS-CoV-2 variant neutralizing antibody in individuals. These findings suggest that the S-IgG antibody is a better supplement to and confirmation of SARS-CoV-2 vaccine immunization.

Objective This study aimed to reveal the urinary and serum metabolic pattern of endometrial cancer (EC) and establish diagnostic models to identify EC from controls, high-risk from low-risk EC, and type II from type I EC. Method This study included 146 EC patients (comprising 79 low-risk and 67 high-risk patients, including 124 type I and 22 type II) and 59 controls. The serum and urine samples were analyzed using ultraperformance liquid chromatography mass spectrometry. Analysis was used to elucidate the distinct metabolites and altered metabolic pathways. Receiver operating characteristic (ROC) analyses were employed to discover and validate the potential biomarker models. Results Serum and urine metabolomes displayed significant differences between EC and controls, with metabolites related to amino acid and nicotinamide metabolisms. The serum and urine panels distinguished these two groups with Area Under the Curve (AUC) of 0.821 and 0.902, respectively. The panel consisting of serum and urine metabolites demonstrated the best predictive ability (AUC = 0.953 and 0.976 in discovering and validation group). In comparing high-risk and low risk EC, differential metabolites were enriched in purine and glutamine metabolism. The AUC values for serum and urine panels were 0.818, and 0.843, respectively. The combined panel exhibited better predictive accuracy (0.881 in discovering group and 0.936 in external validation). In the comparison between type I and type II group, altered folic acid metabolism was identified. The serum, urine and combined panels discriminated these two groups with the AUC of 0.829, 0.913 and 0.922, respectively. Conclusion The combined urine and serum metabolome effectively revealed the metabolic patterns in EC patients, offering valuable diagnostic models for EC diagnosis and classification.

OBJECTIVE:Non-platinum chemotherapy is used in platinum resistant/refractory ovarian cancer patients but offers limited efficacy, especially in those who develop platinum resistance after ≤2 lines of platinum based chemotherapy. This phase II study aimed to evaluate the efficacy and safety of oral niraparib plus etoposide in platinum resistant/refractory ovarian cancer. METHODS:Platinum resistant/refractory ovarian cancer patients after ≤2 lines of platinum based chemotherapy, histologically confirmed as non-mucinous epithelial ovarian cancer, regardless of biomarker status, were eligible. Patients received niraparib with a starting dose of 200 mg/100 mg alternate once a day, and oral etoposide of 50 mg once a day, on days 1-20 of 30 days per cycle for a maximum of 6-8 cycles, followed by niraparib until disease progression or intolerable toxicity. The primary endpoint was investigator assessed progression free survival. RESULTS:29 patients were enrolled from 22 May 2020 to 3 February 2023; 26 patients were included in the efficacy analysis set as per protocol. Median progression free survival was 4.2 months (95% confidence interval (CI) 3.9 to 4.4). Overall response rate was 26.9% (95% CI 8.7 to 45.2). Disease control rate was 57.7% (95% CI 37.3 to 78.0). Overall response rate in patients with a BRCA mutation and homologous recombination deficiency was 50% and 41.7%, respectively. Median progression free survival in patients with primary platinum resistance was 4.5 months (95% CI 3.6 to 5.3). 29 patients were included in the safety analysis set, and 8 (28%) patients experienced treatment related adverse events of grade ≥3. There was no treatment related discontinuation. CONCLUSIONS:Niraparib combined with etoposide showed evidence of antitumor activity in platinum resistant/refractory ovarian cancer after ≤2 lines of platinum based chemotherapy, particularly in patients with a BRCA mutation, homologous recombination deficiency, or primary platinum resistance. This once-a-day oral combination was a convenient option. TRIAL REGISTRATION NUMBER:NCT04217798.

Hand, foot, and mouth disease (HFMD) is caused by the enterovirus family, which includes EV-A71 and more than 10 other members. The disease involves a complex pathological mechanism that includes intricate and finely-tuned interactions between these pathogens and the host. The research on vaccines that can effectively be used to prevent HFMD caused by major pathogens suggested that the viruses presenting with the same structure but different antigenic traits in response to the immune system interactions enable to interact dynamically to cell surface receptors and to lead to similar pathological outcome through diverse mechanisms. This suggests that further understanding of the whole process of signal stimulation by viral antigen molecules and innate immune receptor molecules could improve our recognition about the events of pathological injury to the body and the characterization of antiviral immune responses with phenotypic differences during the pathogenesis. The accumulated data about process of interaction between virus structure and host in molecular level might provide the theoretical and technical support for next generation of vaccine against HFMD for public health initiatives.

Epithelial ovarian cancer (EOC) is a lethal form of gynecological malignancy. Some EOC patients experience relapse after standard primary debulking surgery (PDS) and adjuvant chemotherapy (ACT). Identifying molecular residual disease (MRD) by circulating tumor DNA (ctDNA) detection can timely signal the potential for relapse. However, research on the usage of ctDNA for MRD detection in EOC is limited. Fifty-one EOC patients who received standard PDS and ACT were included. Targeted sequencing based on a panel of 1021 cancer-related genes, along with further validation using Enrich-rare-mutation sequencing, was performed on tumor tissues acquired during PDS and on plasma samples collected before and after PDS/ACT to identify variants reflecting tumor signals. Post-surgery MRD was associated with relapse (Log-rank p = 0.0006) and was identified as an independent prognostic factor (HR, 3.4; 95

Since the mid-twentieth century, hand, foot, and mouth disease (HFMD) was seen as a common cause of viral rash, typically self-limited syndrome in children and adults with classic skin findings. Till the past two decades, HFMD has received new attention because it has led to millions of attacks and several outbreaks across the world. This disease may have completely different clinical epidemiological and etiological characteristics from what was initially believed. Especially, HFMD can be associated with severe complications, such as brainstem encephalitis, meningitis, acute flaccid paralysis (AFP), pulmonary edema (NPE), severe neurological sequelae, and high case-fatality rates. In recent years, it has become a serious health threat and economic burden across the Asia–Pacific region. Historically, outbreaks of HFMD were mainly caused by various enteroviruses. Different pathogens are prevalent in different countries. Vaccines have been developed to provide protection against the most common pathogens in specific countries (e.g., vaccine against enterovirus 71 in China). However, the epidemic of HFMD is complex, such as simultaneous circulation of more than one causative virus and modification of the molecular epidemiology of infectious agents. Awareness of the epidemiological situation and patterns may lead providers to appropriate diagnosis and management.