Objective To explore the feasibility of applying equivalence testing to evaluate the similarity between reference standards and test samples in the biological activity assessment of glucagon like peptide-1(GLP-1) receptor agonist drugs,so as to improve the determination standard of biological activity of various products.Methods Historical release biological activity data from 25 batches of products were collected,and the parameter estimates and standard errors of a four-parameter logistic model were obtained.The slope ratio of test samples to reference standards and dynamic range ratio(ratio of the difference between upper and lower asymptotes of test samples to that of reference standards) were selected as similarity evaluation metrics,and tolerance interval method was used to set equivalence margins for these metrics.Subsequently,biological activity data from 86 additional batches of GLP-1 receptor agonist drugs(57 reference standards with similar biological behaviors to those of test samples and 29 reference standards with non-similar biological behaviors) were analyzed using equivalence testing to determine similarity.For comparison,an F-test was also performed to assess parallelism.The agreement between the two methods was evaluated using Kappa test.Results The equivalence intervals for the slope ratio and dynamic range ratio were 0.64-1.56 and 0.87-1.15,respectively.For the 86 batches,equivalence testing correctly identified 95%(54/57) of truly similar samples as similar and 86%(25/29) of non-similar samples as non-similar.In contrast,the F-test correctly identified 88%(50/57) of parallel samples as parallel and 72%(21/29) of non-parallel samples as non-parallel.Kappa test indicated moderate agreement between the two methods(Kappa=0.417).Conclusion Equivalence testing is suitable for evaluating similarity in the biological activity of GLP-1 receptor agonist drugs and outperforms the F-test in similarity assessment.

Objective To systematically analyze the research status,hotspots,and trends in the field of health-care-associated infection(HAI)in 2020-2025 globally,with a focus on exploring the development strategy of sci-entific and technological talents,and provide data support for talent cultivation,disciplinary layout,and evidence-based decision-making in this field.Methods Web of Science Core Collection was adopted as data source,and HAI-related literatures from January 2020 to January 2025 were retrieved.Bibliometric methods were employed with tools such as VOSviewer and Bibliometrix(R4.4.0)to conduct a cluster analysis on annual publication volume,countries/regions,authors,highly cited literatures,and keywords.The distribution characteristics,collaboration modes,and adaptability to research hotspots of scientific and technological talents in this field were also sorted out.Results A total of 9 566 literatures were included in the analysis.The annual publication volume peaked in 2021 and subsequently entered a high-level platform period with over 1 800 articles per year.China and the United States formed a dual-core research pattern,with Chinese scholars dominating the list of productive authors.Highly cited literatures were predominantly published in 2020-2021,mainly focusing on the clinical characteristics and HAI risks during the early stages of the COVID-19 pandemic.Research hotspots present a dual theme of"clinical epide-miology as well as management of prevention and control"and"antimicrobial resistance mechanisms of pathogens".However,the existing talent system shows deficiency in interdisciplinary adaptability and the depth of international collaboration.Conclusion Over the past five years,global research on HAI has entered a normalized stage,and Chinese scholars play vital role in the dual-core pattern of China and the United States.From the perspective of ta-lents,it is necessary to formulate a development strategy for scientific and technological talents around research hotspots,including improving transnational collaboration mechanisms,innovating interdisciplinary training models,and establishing a dynamic adaptability evaluation system,so as to provide action guidance and policy references for talent development in this field.

To develop a method for determining the antibody-dependent cell-mediated phagocytosis (ADCP) activity of human epidermal growth factor receptor 2 (HER2)-targeted antibody drug conjugate (ADC) based on the reporter gene assay, we established an ADCP activity assay with Jurkat/NFAT/FcγRIIa cells as the effector cells and BT474 as the target cells. Then, the target cell density, the ratio of effector to target cells, the target cell adhesion time, the incubation time for drug administration, and the induction time after adding effector cells were optimized by the method of design of experiment (DOE). The method showed a significant dose-response relationship, which was complied with the four-parameter equation: y=(A-D)/[1+(x/C)^B]+D. The durability ranges of the target cell density, the ratio of effector to target cells, the target cell adhesion time, the incubation time for drug administration, and the induction time after adding effector cells were (2.5-4.0)×10⁵ cells/mL, 3-5, 1.0-2.0 h, 0 h, and 5.0-6.0 h, respectively. The results of the methodological validation showed that the linear equation was y=1.106 8x-0.011 6, r=0.969 2. The established method showed the relative accuracy ranging from -6.59% to 2.98% and the geometric coefficient of variation less than 11% in the intermediate precision test. Furthermore, the method was target-specific. The method was then applied to the determination of ADCP activity of HER2-targeted ADC, demonstrating the result of (103.5±5.7)%. We developed a reporter gene assay for determining the ADCP activity of HER2-targeted ADC and the assay demonstrated high accuracy and good reproducibility, which proposes a highly efficient and approache for evaluating ADCP effect of this HER2-targeted ADC, and also provides a referable technique for characterizing the Fc effector functions of ADCs with diverse targets.
Humans ; Receptor, ErbB-2/immunology* ; Phagocytosis/drug effects* ; Immunoconjugates/immunology* ; Genes, Reporter ; Antibody-Dependent Cell Cytotoxicity ; Jurkat Cells

Objective To establish a prediction model for futile recanalization after mechanical thrombectomy(MT)in acute anterior circulation large vessel occlusion(ACLVO)stroke patients based on multimodal imaging features,and to evaluate its predictive performance.Methods Retrospectively enrolled consecutive ACLVO patients who underwent MT with successful recanalization(modified thrombolysis in cerebral infarction[mTICI]grade≥2b)at the Department of Neurology of Fuyang People's Hospital between June 2023 and December 2024.Demographic and clinical data were collected,including age,gender,hypertension,diabetes mellitus,atrial fibrillation,smoking history,alcohol consumption history,National Institutes of Health stroke scale(NIHSS)score upon admission,intravenous thrombolysis,wake-up stroke,onset-to-puncture time(OPT),puncture-to-recanalization time(PRT),occlusion vessel(internal carotid artery,middle cerebral artery),treatment method(suction thrombectomy,stent thrombectomy,suction+stent thrombectomy).All patients underwent pre-procedural CT perfusion(CTP)+CT angiography(CTA)of the head and neck.Imaging parameters included hypoperfusion(defined as time to peak＞6 s)volume(HPV),core infarct(defined as cerebral blood flow＜30%)volume(CIV),mismatch ratio(MMR;HPV/CIV),and Tan collateral score(poor collaterals:0-1 score,good collaterals:2-3 score).Patients were followed up at 90 days post-procedure via outpatient clinic or re-admission.Patients with a modified Rankin scale(mRS)score≤2 were classified into the effective recanalization group,while those with mRS score≥3 were classified into the futile recanalization group.Imaging variables with statistically significant differences between the futile recanalization and effective recanalization groups were included in multivariate Logistic regression analysis to identify independent predictors of futile recanalization and construct a nomogram model.The predictive value of the model was assessed using the receiver operating characteristic(ROC)curve.Model calibration was evaluated using the Hosmer-Lemeshow test(goodness-of-fit defined as P＞0.50).Results(1)A total of 105 ACLVO patients with successful MT recanalization were included(65 males,40 females,mean age[66±11]years,ranged 31～87 years).There were 60 patients in the effective recanalization group and 45 in the futile recanalization group.Compared to the effective recanalization group,the futile recanalization group had significantly higher age([69±11]years vs.[63±11]years,P=0.012),higher proportion of diabetes mellitus(33.33%[15/45]vs.16.67%[10/60],P=0.047),higher pre-treatment NIHSS score([15.51±2.73]vs.[13.25±2.71],P＜0.01),longer OPT([516.40±192.48]min vs.[322.98±171.22]min,P＜0.01)and PRT([94.96±17.37]min vs.[87.58±15.99]min,P=0.026),larger CIV([74.00±12.76]ml vs.[24.28±14.72]ml,P＜0.01)and HPV([121.43±22.21]ml vs.[91.62±11.34]ml,P＜0.01),smaller MMR([1.65±0.15]vs.[9.42±1.91],P＜0.01),higher 90-day mRS score([3.60±0.54]score vs.[1.22±0.83]score,P＜0.01),and a significantly different distribution of Tan collateral scores(P＜0.01).(2)Multivariate Logistic regression analysis was performed with futile recanalization as the dependent variable,identified the following independent predictors of futile recanalization:HPV(OR,2.042,95%CI 1.296-3.218,P=0.002),CIV(OR,2.373,95%CI 1.315-4.280,P=0.004),MMR(OR,1.758,95%CI 1.135-2.721,P=0.011),and Tan collateral score(OR,5.166,95%CI 2.100-12.651,P＜0.01).(3)A nomogram prediction model for futile recanalization after MT in ACLVO stroke was constructed based on the four imaging parameters as aforementioned.ROC curve analysis demonstrated that the area under the curve for this model in predicting futile recanalization after MT was 0.846(95%CI 0.739-0.912),with a sensitivity of 0.844 and a specificity of 0.817.The calibration curve and the Hosmer-Lemeshow test indicated the goodness-of-fit was high(P=0.617),and the overall stability of the model was good.Conclusion The predictive model for futile recanalization after MT for acute ACLVO constructed base on HPV,CIV,MMR and Tan collateral score facilitates the identification patients with high-risk of futile recanalization.

Objective To systematically analyze the research status,hotspots,and trends in the field of health-care-associated infection(HAI)in 2020-2025 globally,with a focus on exploring the development strategy of sci-entific and technological talents,and provide data support for talent cultivation,disciplinary layout,and evidence-based decision-making in this field.Methods Web of Science Core Collection was adopted as data source,and HAI-related literatures from January 2020 to January 2025 were retrieved.Bibliometric methods were employed with tools such as VOSviewer and Bibliometrix(R4.4.0)to conduct a cluster analysis on annual publication volume,countries/regions,authors,highly cited literatures,and keywords.The distribution characteristics,collaboration modes,and adaptability to research hotspots of scientific and technological talents in this field were also sorted out.Results A total of 9 566 literatures were included in the analysis.The annual publication volume peaked in 2021 and subsequently entered a high-level platform period with over 1 800 articles per year.China and the United States formed a dual-core research pattern,with Chinese scholars dominating the list of productive authors.Highly cited literatures were predominantly published in 2020-2021,mainly focusing on the clinical characteristics and HAI risks during the early stages of the COVID-19 pandemic.Research hotspots present a dual theme of"clinical epide-miology as well as management of prevention and control"and"antimicrobial resistance mechanisms of pathogens".However,the existing talent system shows deficiency in interdisciplinary adaptability and the depth of international collaboration.Conclusion Over the past five years,global research on HAI has entered a normalized stage,and Chinese scholars play vital role in the dual-core pattern of China and the United States.From the perspective of ta-lents,it is necessary to formulate a development strategy for scientific and technological talents around research hotspots,including improving transnational collaboration mechanisms,innovating interdisciplinary training models,and establishing a dynamic adaptability evaluation system,so as to provide action guidance and policy references for talent development in this field.

Objective To establish a prediction model for futile recanalization after mechanical thrombectomy(MT)in acute anterior circulation large vessel occlusion(ACLVO)stroke patients based on multimodal imaging features,and to evaluate its predictive performance.Methods Retrospectively enrolled consecutive ACLVO patients who underwent MT with successful recanalization(modified thrombolysis in cerebral infarction[mTICI]grade≥2b)at the Department of Neurology of Fuyang People's Hospital between June 2023 and December 2024.Demographic and clinical data were collected,including age,gender,hypertension,diabetes mellitus,atrial fibrillation,smoking history,alcohol consumption history,National Institutes of Health stroke scale(NIHSS)score upon admission,intravenous thrombolysis,wake-up stroke,onset-to-puncture time(OPT),puncture-to-recanalization time(PRT),occlusion vessel(internal carotid artery,middle cerebral artery),treatment method(suction thrombectomy,stent thrombectomy,suction+stent thrombectomy).All patients underwent pre-procedural CT perfusion(CTP)+CT angiography(CTA)of the head and neck.Imaging parameters included hypoperfusion(defined as time to peak＞6 s)volume(HPV),core infarct(defined as cerebral blood flow＜30%)volume(CIV),mismatch ratio(MMR;HPV/CIV),and Tan collateral score(poor collaterals:0-1 score,good collaterals:2-3 score).Patients were followed up at 90 days post-procedure via outpatient clinic or re-admission.Patients with a modified Rankin scale(mRS)score≤2 were classified into the effective recanalization group,while those with mRS score≥3 were classified into the futile recanalization group.Imaging variables with statistically significant differences between the futile recanalization and effective recanalization groups were included in multivariate Logistic regression analysis to identify independent predictors of futile recanalization and construct a nomogram model.The predictive value of the model was assessed using the receiver operating characteristic(ROC)curve.Model calibration was evaluated using the Hosmer-Lemeshow test(goodness-of-fit defined as P＞0.50).Results(1)A total of 105 ACLVO patients with successful MT recanalization were included(65 males,40 females,mean age[66±11]years,ranged 31～87 years).There were 60 patients in the effective recanalization group and 45 in the futile recanalization group.Compared to the effective recanalization group,the futile recanalization group had significantly higher age([69±11]years vs.[63±11]years,P=0.012),higher proportion of diabetes mellitus(33.33%[15/45]vs.16.67%[10/60],P=0.047),higher pre-treatment NIHSS score([15.51±2.73]vs.[13.25±2.71],P＜0.01),longer OPT([516.40±192.48]min vs.[322.98±171.22]min,P＜0.01)and PRT([94.96±17.37]min vs.[87.58±15.99]min,P=0.026),larger CIV([74.00±12.76]ml vs.[24.28±14.72]ml,P＜0.01)and HPV([121.43±22.21]ml vs.[91.62±11.34]ml,P＜0.01),smaller MMR([1.65±0.15]vs.[9.42±1.91],P＜0.01),higher 90-day mRS score([3.60±0.54]score vs.[1.22±0.83]score,P＜0.01),and a significantly different distribution of Tan collateral scores(P＜0.01).(2)Multivariate Logistic regression analysis was performed with futile recanalization as the dependent variable,identified the following independent predictors of futile recanalization:HPV(OR,2.042,95%CI 1.296-3.218,P=0.002),CIV(OR,2.373,95%CI 1.315-4.280,P=0.004),MMR(OR,1.758,95%CI 1.135-2.721,P=0.011),and Tan collateral score(OR,5.166,95%CI 2.100-12.651,P＜0.01).(3)A nomogram prediction model for futile recanalization after MT in ACLVO stroke was constructed based on the four imaging parameters as aforementioned.ROC curve analysis demonstrated that the area under the curve for this model in predicting futile recanalization after MT was 0.846(95%CI 0.739-0.912),with a sensitivity of 0.844 and a specificity of 0.817.The calibration curve and the Hosmer-Lemeshow test indicated the goodness-of-fit was high(P=0.617),and the overall stability of the model was good.Conclusion The predictive model for futile recanalization after MT for acute ACLVO constructed base on HPV,CIV,MMR and Tan collateral score facilitates the identification patients with high-risk of futile recanalization.

Objective To explore the possible mechanism of apoptosis induced by SARS-CoV-2 targeting cyclophilin A(CypA)/CD147 receptor pathway.Methods The pEncmv-SARS-CoV-2 S-3xflag recombinant plasmid was constructed and transfected into H9C2 cells.Apoptosis was analyzed by TUNEL assay,flow cytometry and DNA ladder.The interaction between CD147 and SARS-CoV-2 S protein was verified by CoIP.The expressions of apoptosis-related proteins were detected by Western blotting.Results Compared with the control group,TUNEL assay and flow cytometry showed that apoptosis was significantly increased in transfection group(P＜0.05),and DNA ladder analysis showed that DNA degradation was obvious.The expression levels of Caspase12 and DR3 were significantly increased(P＜0.001),the expression level of FAS was slightly increased(P＜0.01),while the expression levels of TRF1 and DR4 were decreased(P＜0.01).Conclusion SARS-CoV-2 S protein targets CypA/CD147 receptor to invade host cells,activate endoplasmic network pathway(Caspase12)and death receptor pathway(DR3,FAS),and induce apoptosis of cardiomyocytes through endogenous and exogenous apoptotic pathways.

Objective To analyze the correlation of inflammatory markers C-reactive proten(CRP),serum amyloid protein A(SAA),interleukin-6(IL-6)levels and subjective sleep characteristics in patients with first-episode acute cerebral infarction.Methods A total of 113 patients with first-episode cerebral infarction admitted to the Department of Neurology,the People's Hospital of Fuyang from March 2022 to April 2023 were prospectively and continuously selected as subjects.According to the Pittsburgh sleep quality index(PSQI),they were divided into insomnia group(PSQI＞7 points)and non-insomnia group(PSQI ≤7 points).General demo-graphic data and differences in CRP,SAA,IL-6 levels,Hamilton anxiety scale(HAMA)and Hamilton depression scale(HAMD)scores were compared between the two groups.Partial correlation analysis was used to analyze the correlation between three serum markers and PSQI effect factors.Results There were no significant differences in age,gender,baseline NIHSS score and mRS score between the two groups(P＞0.05).HAMD scores(z=-3.993,P＜0.001),HAMA scores(z=-3.806,P＜0.001),CRP,IL-6,SAA(P＜0.001)in insomnia group were significantly higher than those in non-insomnia group.The history of hyperlipidemia between the two groups was statistically significant(z=5.913,P=0.015).Multivariate Logisitic regression analysis showed that CRP(OR=1.55,P＜0.01),HAMD scores,HAMA scores and hyperlipidemia were independent risk factors for chronic insomnia in patients with first-episode cerebral infarction,and HAMD scores had a greater effect than HAMA scores(OR:1.10 vs 1.04).Partial correlation analysis showed that IL-6 and CRP levels were significantly correlated with the total score of PSQI(P＜0.05),while SAA was not significantly correla-ted with the total score of PSQI(P＞0.05).IL-6 level was positively correlated with sleep quality(r=0.231)sleep efficiency(r=0.322)and sleep duration(r=0.221).SAA level was positively correlated with sleep efficiency(r=0.242),while CRP level was posi-tively correlated with sleep latency(r=0.194),sleep duration(r=0.247)and sleep efficiency(r=0.225).Conclusion The inflam-matory markers CRP,IL-6 and SAA levels were elevated in the patients with first-episode cerebral infarction accompanied by insomnia,which were correlated with the severity of insomnia.The correlation between CRP and IL-6 levels and sleep characteristics was consistent with each other.

Objective: To explore the hemorrhagic transformation(HT) and its effect on prognosis in patients with acute ischemic stroke(AIS) after mechanical thrombectomy(MT). Methods: A total of 114 patients with AIS received MT were enrolled. The modified Rankin Scale was used to evaluate the clinical outcome at 90 days of onset(0-2 points were good prognosis; 3-6 points were poor prognosis). The patients were divided into HT group(n=25) and non-HT group(n=89) according to their HT conditions. Binomial Logistic regression analysis was performed to determine the vascular risk factors of HT after MT and the effect of HT on prognosis. Results: Among 114 patients, there were 25 cases of HT and 89 cases of non HT. The proportion of patients with diabetes in HT group was significantly higher than that in non-HT group. The NIHSS score of HT group at discharge was significantly higher than that in non-HT group. The proportion with good prognosis at 90 days in HT group was significantly lower than that in non-HT group(allP<0.05). Binomial Logistic regression analysis showed that diabetes, high levels of cholesterol and smoking were the major vascular risk factors for HT after thrombectomy(allP<0.05). HT was an important factor affecting the poor prognosis after arterial thrombectomy(P=0.026). Conclusion Diabetes, high levels of cholesterol and smoking are the main risk factors of HT after MT for AIS. HT is an independent risk factor for poor prognosis after MT.

Lipopolysaccharide(LPS),lipteichoic acid(LTA) and zymosan can be recognized by Toll-like receptors (TLRs) ex-pressing in the inflammatory cells membrane,such as macrophages,monocytes and endothelial cells,and then induce the secretion of pyrogenic cytokines followed by triggering inflammatory reaction. The signaling pathway mechanisms were summarized, including the extracellular signaling cascades from pyrogens to individual TLRs,and intracellular signaling cascades from TLRs to nucleus.