Objective: To validate the performance of the Procleix UltrioPlex E assay (Grifols, Spain) on the Procleix Panther automated nucleic acid detection platform, which employs the TMA method to simultaneously detect HIV-1/HIV-2/HCV/HBV/HEV viruses, and to evaluate its value for screening transfusion-associated infectious diseases. Methods: In accordance with the requirements of ISO15189"Application of the Guidelines for the Accreditation of Quality and Capabilities of Medical Laboratories in the Field of Molecular Diagnostics (CNAS-CL02-A009: 2018)", "Guidelines for Performance Validation of Molecular Diagnostic Testing Procedures (CNAS-GL039: 2019)", and the "Technical Operating Procedures for Blood Banks (2019 Edition)", this study validated the reagent's performance in terms of analytical sensitivity validation, performance consistency validation, interference resistance, and cross-contamination resistance. Results: Probit analysis revealed that the 95% detection limits (95% confidence interval) for HBV, HCV, HIV, and HEV were 2.0 IU/mL, 1.5 IU/mL, 18.0 IU/mL and 3.7 IU/mL, respectively, which were consistent with the minimum detection limits stated in the kit's package insert and were comparable to the Procleix Ultrio Elite kit. Both kits were used to test the performance validation serum plate simultaneously, yielding results consistent with the serum plate (Kappa=1), indicating stable performance. Detection of medium-and low-concentration lipemia and weakly positive hemolysis samples demonstrated good interference resistance. Cross-contamination performance validation showed that the kit exhibited excellent cross-contamination resistance. Conclusion: The Procleix UltrioPlex E nucleic acid detection kit enables combined detection of HIV-1, HIV-2, HCV, HBV, and HEV, allowing single-test screening for multiple viruses in donor blood. The kit's analytical performance is stable and meets basic laboratory requirements, making it suitable for screening transfusion-associated infectious diseases in blood banks.

Escherichia coli(E.coli)is a key pathogenic bacterium responsible for postpartum endo-metritis,with its colonization in the reproductive tract closely associated with endometrial damage and disruption of the ovarian cycle.This ultimately leads to infertility,causing significant economic losses to the dairy industry.Macrophages play a pivotal role in the inflammatory response.This study aims to investigate the mRNA expression profile of bovine bone marrow-derived macropha-ges following E.coli infection using RNA sequencing(RNA-seq)technology.Additionally,it seeks to identify the biological functions and signaling pathways of differentially expressed genes(DEGs)through Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.The results demonstrated that E.coli infection induced differential expression of 4 522 genes,with 2 141 upregulated and 2 381 downregulated.These genes were primarily asso-ciated with inflammatory responses,where TLR2,TLR4,NLRP3,and PTGS2 played pivotal roles.PGD2 synthesis was mediated by TLR2,TLR4,and NLRP3.Transcriptome sequencing of bovine bone marrow-derived macrophages infected with E.coli and treated with a PGD2 inhibitor revealed a marked downregulation of TLR2 and TLR4 gene expression.qPCR validation results were highly consistent with the RNA-seq findings.This study elucidates the interactive regulatory roles of TLR2,TLR4,and NLRP3 in conjunction with PGD2,which collectively modulate bovine endome-tritis.These findings offer significant molecular insights that enhance our understanding of the pathological mechanisms underlying bovine endometritis,thereby informing its prevention and treatment strategies.

Escherichia coli(E.coli)is a key pathogenic bacterium responsible for postpartum endo-metritis,with its colonization in the reproductive tract closely associated with endometrial damage and disruption of the ovarian cycle.This ultimately leads to infertility,causing significant economic losses to the dairy industry.Macrophages play a pivotal role in the inflammatory response.This study aims to investigate the mRNA expression profile of bovine bone marrow-derived macropha-ges following E.coli infection using RNA sequencing(RNA-seq)technology.Additionally,it seeks to identify the biological functions and signaling pathways of differentially expressed genes(DEGs)through Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.The results demonstrated that E.coli infection induced differential expression of 4 522 genes,with 2 141 upregulated and 2 381 downregulated.These genes were primarily asso-ciated with inflammatory responses,where TLR2,TLR4,NLRP3,and PTGS2 played pivotal roles.PGD2 synthesis was mediated by TLR2,TLR4,and NLRP3.Transcriptome sequencing of bovine bone marrow-derived macrophages infected with E.coli and treated with a PGD2 inhibitor revealed a marked downregulation of TLR2 and TLR4 gene expression.qPCR validation results were highly consistent with the RNA-seq findings.This study elucidates the interactive regulatory roles of TLR2,TLR4,and NLRP3 in conjunction with PGD2,which collectively modulate bovine endome-tritis.These findings offer significant molecular insights that enhance our understanding of the pathological mechanisms underlying bovine endometritis,thereby informing its prevention and treatment strategies.

Objective:Bidirectional causal associations of 41 cytokines with atopic dermatitis(AD)were explored based on a Mendelian randomization(MR)approach.Methods:Pooled data from genome wide association study(GWAS)of 41 cytokines and AD were utilized for instrumental variable(IV)screening,and single nucleotide polymorphism(SNP)affecting the results of MR analyses was excluded by the MR-PRESSO outlier test as well as by the MR Steiger filtering method.Two-sample bidirectional MR analyses were performed using inverse variance weighting(IVW),MR-Egger regression,and weighted median methods(WM).MR-Egger intercept term test and Cochran's Q test were performed to test the pleiotropy and heterogeneity of IV,and MR results were visu-alized by scatterplots,funnel plots,and leave-one-out plots.Results:Forward MR analysis showed that MIG(IVW:OR=0.89;95%CI:0.81～0.97;P=0.006)reduced the risk of AD development.In contrast,IL-5(IVW:OR=1.17;95%CI:1.01～1.36;P=0.042)and IL-18(MR Egger:OR=1.17;95%CI:1.03～1.33;P=0.030)increased the risk of AD development.Inverse MR analysis showed a potential causal association between AD and increased MIG(IVW:Beta=0.10;95%CI:0.02～0.17;P=0.014).None of the sensitivity analyses indicated pleiotropy and heterogeneity of the included IV.Conclusion:MIG may be an important marker in the progression of AD with a potential bidirectional causal association with risk of morbidity.IL-5 and IL-18 have a potential positive causal association for AD.

Objective·To develop an early diagnostic model for small cell lung cancer(SCLC)based on differences in serum metabolite expression profiles between patients with SCLC and those with benign pulmonary diseases,using machine learning algorithms.Methods·Serum samples were collected from 29 SCLC patients and 67 patients with benign lung diseases at Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,as the training cohort.An independent external validation cohort included 20 SCLC patients and 40 patients with benign lung diseases from Gansu Provincial Cancer Hospital.A total of 69 serum metabolites were quantitatively analyzed using liquid chromatography-tandem mass spectrometry(LC-MS/MS).The XGBoost Classifier was employed to rank metabolite importance,and a forward feature selection strategy based on XGBoost was used to identify a subset of key metabolites.Diagnostic models were constructed using AdaBoost,random forest(RF),and light gradient boosting machine(LGBM)algorithms.Model performance was assessed using receiver operating characteristic(ROC)curves and the area under the curve(AUC),and validated on the external test cohort.Results·Principal component analysis(PCA)and orthogonal projections to latent structures-discriminant analysis(OPLS-DA)of the training cohort revealed distinct metabolic profiles between SCLC and benign lung disease patients.Based on feature importance rankings,six key metabolites were selected to construct the MTB-6 diagnostic model.Among the models,AdaBoost achieved the best performance,with an AUC of 0.943,sensitivity of 75.0%,and specificity of 90.9%in the training cohort.In the external test cohort,the model demonstrated robust performance with an AUC of 0.921,sensitivity of 80.0%,and specificity of 87.5%.Conclusion·The MTB-6 model,based on six serum metabolites and the AdaBoost algorithm,exhibits excellent diagnostic performance and holds potential for the differential diagnosis of SCLC and benign pulmonary diseases.

BACKGROUND: Durvalumab after chemoradiotherapy (CRT) failed to bring survival benefits to patients with epidermal growth factor receptor ( EGFR ) mutations in PACIFIC study (evaluating durvalumab in patients with stage III, unresectable NSCLC who did not have disease progression after concurrent chemoradiotherapy). We aimed to explore whether locally advanced inoperable patients with EGFR mutations benefit from tyrosine kinase inhibitors (TKIs) and the optimal treatment regimen. METHODS: We searched the PubMed, Embase, the Cochrane Central Register of Controlled Trials, and ClinicalTrials.gov databases from inception to December 31, 2022 and performed a meta-analysis based on a Bayesian framework, with progression-free survival (PFS) and overall survival (OS) as the primary endpoints. RESULTS: A total of 1156 patients were identified in 16 studies that included 6 treatment measures, including CRT, CRT followed by durvalumab (CRT-Durva), TKI monotherapy, radiotherapy combined with TKI (RT-TKI), CRT combined with TKI (CRT-TKI), and TKI combined with durvalumab (TKI-Durva). The PFS of patients treated with TKI-containing regimens was significantly longer than that of patients treated with TKI-free regimens (hazard ratio [HR] = 0.37, 95% confidence interval [CI], 0.20-0.66). The PFS of TKI monotherapy was significantly longer than that of CRT (HR = 0.66, 95% CI, 0.50-0.87) but shorter than RT-TKI (HR = 1.78, 95% CI, 1.17-2.67). Furthermore, the PFS of RT-TKI or CRT-TKI were both significantly longer than that of CRT or CRT-Durva. RT-TKI ranked first in the Bayesian ranking, with the longest OS (60.8 months, 95% CI = 37.2-84.3 months) and the longest PFS (21.5 months, 95% CI, 15.4-27.5 months) in integrated analysis. CONCLUSIONS: For unresectable stage III EGFR mutant NSCLC, RT and TKI are both essential. Based on the current evidence, RT-TKI brings a superior survival advantage, while CRT-TKI needs further estimation. Large randomized clinical trials are urgently needed to explore the appropriate application sequences of TKI, radiotherapy, and chemotherapy. REGISTRATION PROSPERO; https://www.crd.york.ac.uk/PROSPERO/ ; No. CRD42022298490.
Humans ; Carcinoma, Non-Small-Cell Lung/therapy* ; ErbB Receptors/genetics* ; Lung Neoplasms/drug therapy* ; Mutation/genetics* ; Protein Kinase Inhibitors/therapeutic use* ; Chemoradiotherapy ; Antibodies, Monoclonal/therapeutic use*

ObjectiveTo investigate the triglyceride-glucose (TyG) index and the level of serum homocysteine (Hcy) in association with the incidence of stroke in type 2 diabetes mellitus (T2DM) patients. MethodsBased on the chronic disease risk factor surveillance cohort in Pudong New Area, Shanghai, excluding those with stroke in baseline survey, T2DM patients who joined the cohort from January 2016 to October 2020 were selected as the research subjects. During the follow-up period, a total of 318 new-onset ischemic stroke patients were selected as the case group, and a total of 318 individuals matched by gender without stroke were selected as the control group. The Cox proportional hazards regression model was used to adjust for confounding factors and explore the serum TyG index and the Hcy biochemical indicator in association with the risk of stroke. ResultsThe Cox proportional hazards regression results showed that after adjusting for confounding factors, the risk of stroke in T2DM patients with 10 μmol·L⁻¹-¹ and Hcy>15 μmol·L⁻¹ was 1.532 times (95%CI: 1.017‒2.309 times, P=0.041) and 1.738 times (95%CI: 1.119‒2.699 times, P=0.014) higher respectively, compared to T2DM patients with Hcy≤10 μmol·L⁻¹. T2DM patients with TyG>9.074 had 1.396 times higher risk of stroke (95%CI: 1.091‒1.787, P=0.008) compared to those with TyG≤9.074. The study found that urea and α1-antitrypsin (α1-AT) were independent risk factors for the incidence of stroke in T2DM patients. For every unit increase in urea andα1-AT, the risk of stroke in T2DM patients increased by 233.6% (HR=3.336, 95%CI: 1.588‒7.009, P=0.001) and 11.3% (HR=1.113, 95%CI: 1.028‒1.205, P=0.008), respectively. Cumulative risk curves showed that Hcy>10 μmol·L⁻¹ and TyG>9.074 accelerated the time to stroke occurrence in T2DM patients. ConclusionElevated levels of Hcy>10 μmol·L⁻¹ and a higher TyG index are risk factors for stroke in T2DM patients. Effective interventions for Hcy and TyG should be conducted for diabetic patients to reduce the incidence of stroke.

INTRODUCTION: Paediatric sexual assault (SA) victims should be assessed for post-exposure prophylaxis (PEP) to mitigate the risk of sexually transmitted infections (STIs). We describe the clinical characteristics of children and young persons (CYPs) presenting with SA at KK Women's and Children's Hospital in Singapore, viral PEP (human immunodeficiency virus [HIV] and hepatitis B virus [HBV]) prescribing practices, and STI evaluation at follow-up. METHOD: Medical records of CYPs ≤16 years who presented with SA between January 2022 and August 2023 were reviewed, including assault and assailant characteristics, baseline and follow-up STI screening, PEP prescription, adherence and follow-up attendance. CYPs with SA in the preceding 72 hours by HIV-positive or HIV-status unknown assailants with high-risk characteris-tics were eligible for HIV PEP. RESULTS: We analysed 278 CYPs who made 292 SA visits. There were 40 (13.7%) CYPs eligible for HIV PEP, of whom 29 (82.9%) received it. Among those tested at baseline, 9% and 34.9% of CYPs tested positive for Chlamydia trachomatis and Gardnerella vaginalis, respectively. None tested positive for Neisseria gonorrhoeae, Trichomonas vaginalis, HIV, HBV or hepatitis C. Majority of CYPs tested were HBV non-immune (n=167, 67.6%); only 77 (46.1%) received the vaccine. Out of 27 CYPs eligible for HBV PEP with immunoglobulin, only 21 (77.7%) received immunoglobulin. A total of 37 CYPs received HIV PEP, including 8 who were retrospectively deemed ineligible. Only 10 (27%) completed the course. Overall, 153 (57.7%) CYPs attended follow-up, and none seroconverted for HIV or HBV. CONCLUSION We report suboptimal rates of HBV post-exposure vaccination, and low compliance to HIV PEP and follow-up among paediatric SA victims. Factors contri-buting to poor compliance should be examined to optimise care for this vulnerable population.
Humans ; Post-Exposure Prophylaxis/methods* ; Female ; Child ; Adolescent ; Singapore/epidemiology* ; HIV Infections/prevention & control* ; Male ; Sexually Transmitted Diseases/epidemiology* ; Retrospective Studies ; Hepatitis B/prevention & control* ; Follow-Up Studies ; Child, Preschool ; Sex Offenses/statistics & numerical data* ; Child Abuse, Sexual

Ischemic stroke is more common in the elderly, characterized by rapid onset, high incidence, high disability rate, and high mortality rate.It often causes severe cellular dysfunction and brain damage, imposing a significant burden on families and society.Death-associated protein kinase 1(DAPK1)is a calcium/calmodulin(Ca 2+ /CaM)-dependent serine/threonine(Ser/Thr)protein kinase that plays a crucial role in regulating brain damage caused by ischemic stroke.Studies have shown that DAPK1 activity is regulated by phosphorylation and mediates the pathophysiological processes of ischemic stroke through multiple signaling pathways, including DAPK1-N-methyl-D-aspartate(DAPK1-NMDA), DAPK1-NMDA), DAPK1-tumor suppressor protein p53(DAPK1-p53), DAPK1-microtubule-associated protein Tau(DAPK1-Tau), and DAPK1-dangerous signaling(DAPK1-DANGER), among others.Blocking these signaling pathways can significantly improve neuronal damage following ischemic stroke.This article reviews the related signaling pathways, mechanisms of action, and progress in targeted drug development of DAPK1 in ischemic stroke, aiming to provide references for future research and treatment of ischemic stroke.