The continuous emergence of SARS-CoV-2 variants as well as other potential future coronavirus has challenged the effectiveness of current COVID-19 vaccines. Therefore, there remains a need for alternative antivirals that target processes less susceptible to mutations, such as the formation of six-helix bundle (6-HB) during the viral fusion step of host cell entry. In this study, a novel high-throughput screening (HTS) assay employing a yeast-two-hybrid (Y2H) system was established to identify inhibitors of HR1/HR2 interaction. The compound IMB-9C, which achieved single-digit micromolar inhibition of SARS-CoV-2 and its Omicron variants with low cytotoxicity, was selected. IMB-9C effectively blocks the HR1/HR2 interaction in vitro and inhibits SARS-CoV-2-S-mediated cell-cell fusion. It binds to both HR1 and HR2 through non-covalent interaction and influences the secondary structure of HR1/HR2 complex. In addition, virtual docking and site-mutagenesis results suggest that amino acid residues A930, I931, K933, T941, and L945 are critical for IMB-9C binding to HR1. Collectively, in this study, we have developed a novel screening method for HR1/HR2 interaction inhibitors and identified IMB-9C as a potential antiviral small molecule against COVID-19 and its variants.

Aim To detect the changes of serum cardiotrophin-1(CT-1)and angiopoietin-like protein 3(ANGPTL3)levels in patients with coronary heart disease(CHD)complicated with heart failure(HF)after percutaneous coronary intervention(PCI),and analyze their relationship with prognosis.Methods 199 patients with CHD compli-cated with HF who underwent PCI in the Second Affiliated Hospital of Zhengzhou University from March 2022 to March 2023 were selected.The serum CT-1 and ANGPTL3 levels of patients with different New York Heart Association(NYHA)cardiac function grades were compared before surgery.The prognosis was followed up after PCI,and the pa-tients who had major adverse cardiovascular event(MACE)were included in the poor prognosis group,and the rest were included in the good prognosis group.The general data and serum CT-1 and ANGPTL3 levels were compared between the poor prognosis group and the good prognosis group.Logistic regression model was used to analyze the influencing factors of poor prognosis after surgery in patients with CHD and HF.The predictive value of serum CT-1 and ANGPTL3 alone and in combination were analyzed.Results Compared with the patients with cardiac function grade Ⅰ,the serum CT-1 and ANGPTL3 levels of the patients with cardiac function grade Ⅱ,Ⅲ and Ⅳ were increased(P＜0.05).Compared with the patients with cardiac function grade Ⅱ,the serum CT-1 and ANGPTL3 levels of the patients with cardiac function grade Ⅲ and Ⅳ were increased(P＜0.05).Compared with the patients with cardiac function grade Ⅲ,the serum CT-1 and ANGPTL3 levels of the patients with acrdiac function grade Ⅳ were increased(P＜0.05).Spearman correlation a-nalysis showed that the serum CT-1 and ANGPTL3 levels were positively correlated with NYHA cardiac function grade(r=0.518,95％CI:0.408～0.613,P＜0.001,r=0.737,95％CI:0.666～0.794,P＜0.001).The poor prognosis rate of patients was 17.93％.Compared with the good prognosis group,the serum CT-1 and ANGPTL3 levels of the poor progno-sis group were increased(P＜0.05).Logistic regression model analysis showed that smoking,diabetes,lesion vessel number ≥3,irregular medication outside the hospital,serum CT-1 and ANGPTL3 levels were the influencing factors of poor prognosis in patients with CHD complicated with HF(P＜0.05).ROC curve analysis showed that the sensitivity and area under the curve(AUC)of combined serum CT-1 and ANGPTL3 levels for predicting poor prognosis of patients with CHD complicated with HF were higher than those of either marker alone,while the specificity was basically similar to that of sin-gle-marker prediction.Conclusion Serum CT-1 and ANGPTL3 levels are abnormally elevated in patients with CHD complicated with HF after PCI,and are closely related to the cardiac function and prognosis.

ObjectiveTo investigate the effect and underlying mechanism of the Wulao Qisun prescription on pathological new bone formation in ankylosing spondylitis （AS）. MethodsSynovial fibroblasts were isolated from the hip joints of AS patients and observed under a microscope to assess cell morphology. The cells were identified using immunofluorescence staining. The isolated AS fibroblasts were divided into blank group， low drug-containing serum group， medium drug-containing serum group， high drug-containing serum group， and positive drug group. After drug intervention， cell proliferation was measured using the cell counting kit-8 （CCK-8） assay to observe fibroblast growth and determine the optimal intervention time. Alkaline phosphatase （ALP） activity was measured using the alkaline phosphatase assay. Protein expression of osteocalcin （OCN）， osteopontin （OPN）， and runt-related transcription factor 2 （Runx2） was detected by Western blot. The mRNA expression levels of Wnt5a， β-catenin， and Dickkopf-1 （DKK-1） were measured by real-time quantitative polymerase chain reaction （Real-time PCR）. ResultsCompared with the blank group， each drug-containing serum group of Wulao Qisun prescription and the positive drug group inhibited the proliferation of AS fibroblasts and reduced ALP expression （P<0.01）. Compared with the blank group， the low drug-containing serum group of Wulao Qisun prescription downregulated β-catenin mRNA expression （P<0.05）. The medium and high drug-containing serum groups and the positive drug group significantly downregulated Wnt5a and β-catenin mRNA expression （P<0.05， P<0.01）， with the positive drug group showing the most pronounced effect （P<0.01）. The high drug-containing serum group and the positive drug group significantly upregulated DKK-1 mRNA expression （P<0.01）. Compared with the blank group， the low drug-containing serum group of Wulao Qisun prescription inhibited the expression of OPN and Runx2 proteins （P<0.05， P<0.01）， while the medium and high drug-containing serum groups and the positive drug group inhibited the expression of OCN， OPN， and Runx2 proteins （P<0.05， P<0.01）. ConclusionThe Wulao Qisun prescription can inhibit the proliferation and osteogenic differentiation of AS fibroblasts， thereby delaying the formation of pathological new bone in AS. The possible mechanism involves the regulation of Wnt/β-catenin-related gene expression， further inhibiting the transcription of downstream target genes.

Objective To explore the biological functions of pyroptosis-related genes in pneumonic plague using bioinformatics methods,and to evaluate their potential applicability as diagnostic markers.Methods The pneumonic plague-related dataset GSE220123 was retrieved from the Gene Expression Omnibus(GEO)database and screened for differentially expressed pyroptosis-related genes(DE-PRGs).The functions of DE-PRGs were studied via Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses,and immune infiltration analysis.The hub genes were identified via protein-protein interaction(PPI)network analysis,and further screened for key genes with sustained high expression characteristics based on differential expression analysis.The relative expression levels of the key genes were verified using the reverse transcription real-time quantitative PCR(qPCR)method.Results A total of 17 DE-PRGs were screened,and PPI network analysis revealed 7 Hub genes.Among them,Casp4 continued to be up-regulated during the course of pneumonic plague.The results of reverse transcription qPCR were consistent with the those of bioinformatic analyses.Conclusion DE-PRGs play a crucial role in the immune response of pneumonic plague,especially Casp4,which has significant applications as a diagnostic biomarker and potential therapeutic target for pneumonic plague.

Objective : To identify autophagy-related genes involved in pulmonary infection caused by the highly drug-resistant and virulent methicillin-resistant Staphylococcus aureus strain USA300 ( USA300) ，and to explore the underlying molecular mechanisms ， thereby providing potential targets for immunotherapy． Methods: The GSE220943 dataset of a USA300-induced pulmonary infection mouse model was obtained from the GEO database． Differentially expressed genes ( DEGs ) were identified using the DESeq2 package． Autophagy-related genes ( ARGs) were retrieved from the MSigDB and Autophagy databases．Weighted gene co-expression network analysis ( WGCNA) was performed to construct gene co-expression modules．Genes overlapping among DEGs，ARGs，and WGCNA modules were identified as autophagy-related DEGs．Gene Ontology ( GO) enrichment analysis was con- ducted using the clusterProfiler R package，while Kyoto Encyclopedia of Genes and Genomes ( KEGG) pathway en- richment analysis was performed via the Metascape platform．Immune cell infiltration was analyzed using the Immu- CellAI-mouse website．A protein - protein interaction ( PPI) network was constructed using the STRING database， and hub genes were identified through topological analysis in Cytoscape． Receiver operating characteristic curve ( ROC) curves were plotted via the website https: / /www．bioinformatics．com．cn． Finally，key gene expression was validated in mouse lung tissues by real-time quantitative reverse transcription PCR ( RT-qPCR) ． Results: A total of 6 135，4 075，3 680，and 2 342 differentially expressed genes ( DEGs) were identified at 12，24，48，and 96 hours post-infection，respectively．By integrating DEGs，autophagy-related genes ( ARGs) ，and WGCNA mod- ules，19 autophagy-related DEGs were identified． GO and KEGG enrichment analyses indicated that these genes were mainly involved in CD4 + T cell activation and regulation，innate immune responses，and autophagosome mem- brane formation．Immune infiltration analysis revealed that innate immune cells such as neutrophils and dendritic cells predominated during the early phase of infection，while γδ T cells and M2 macrophages became more promi- nent in the later stages．PPI network analysis identified 12 hub autophagy-related genes，among which three upreg- ulated key genes ( Eif2ak2，Ikbke，and Nfkbiz) were further confirmed．The area under the ROC curve for all three genes was 1. 000．RT-qPCR validation demonstrated significantly elevated expression of these three genes in lung tissues at 24 hours post-infection ( all P＜0. 05) ． Conclusion Eif2ak2，Ikbke，and Nfkbiz may be involved in the pulmonary infection caused by USA300 by promoting autophagy and hold promise as potential targets for immuno- therapy．

Objective To reveal the detailed histological characteristics of pyramidal neuron cell bodies and their axonal projections along the corticospinal tract in the primary motor cortex(M1)of the brain,by using the biotinylated dextran amine(BDA)neural tracing technique combined with panoramic and local microscopic imaging technologies.Methods A total of 100 nL of 10％BDA(10,000 molecular weight)was injected into M1 region using stereotaxic system.The distribution of BDA labeling along the corticospinal tract was continuously tracked with panoramic tissue scanning analysis system.Detailed observations of the histological characteristics of BDA labeling were carried out with laser confocal microscope.Results It is more convenient to observe the overall distribution of BDA neural labeling by using the panoramic tissue scanning analysis system.Around the injection site in M1,the BDA labeling was shown in the somas of pyramidal neurons in layer V.In the M1 region corresponding to the contralateral site of the injection site and ipsilateral primary sensory cortex,BDA showed predominantly the anterograde labeled nerve fibers accompanied by a few retrograde labeled neurons.Besides,BDA labeled nerve fibers-including bundles and terminals-projecting to regions such as the ipsilateral striatum,thalamus,internal capsule,cerebral peduncle,and pons,and further reaching the contralateral spinal cord via the brainstem pyramidal decussation.Confocal microscopy and its 3D reconstruction system facilitated detailed analysis of the local microscopic features of BDA labeling,revealing retrograde labeled neuron cell bodies,dendrites and their spines,as well as anterograde labeled nerve fibers and their terminals.Conclusions These findings demonstrated that the integration of traditional BDA neural tracing with panoramic tissue scanning analysis and confocal microscopy provided an effective approach to the observation and analysis of long-projection neural circuits from panoramic to local perspectives,with broad application prospects.

Aim To detect the changes of serum cardiotrophin-1(CT-1)and angiopoietin-like protein 3(ANGPTL3)levels in patients with coronary heart disease(CHD)complicated with heart failure(HF)after percutaneous coronary intervention(PCI),and analyze their relationship with prognosis.Methods 199 patients with CHD compli-cated with HF who underwent PCI in the Second Affiliated Hospital of Zhengzhou University from March 2022 to March 2023 were selected.The serum CT-1 and ANGPTL3 levels of patients with different New York Heart Association(NYHA)cardiac function grades were compared before surgery.The prognosis was followed up after PCI,and the pa-tients who had major adverse cardiovascular event(MACE)were included in the poor prognosis group,and the rest were included in the good prognosis group.The general data and serum CT-1 and ANGPTL3 levels were compared between the poor prognosis group and the good prognosis group.Logistic regression model was used to analyze the influencing factors of poor prognosis after surgery in patients with CHD and HF.The predictive value of serum CT-1 and ANGPTL3 alone and in combination were analyzed.Results Compared with the patients with cardiac function grade Ⅰ,the serum CT-1 and ANGPTL3 levels of the patients with cardiac function grade Ⅱ,Ⅲ and Ⅳ were increased(P＜0.05).Compared with the patients with cardiac function grade Ⅱ,the serum CT-1 and ANGPTL3 levels of the patients with cardiac function grade Ⅲ and Ⅳ were increased(P＜0.05).Compared with the patients with cardiac function grade Ⅲ,the serum CT-1 and ANGPTL3 levels of the patients with acrdiac function grade Ⅳ were increased(P＜0.05).Spearman correlation a-nalysis showed that the serum CT-1 and ANGPTL3 levels were positively correlated with NYHA cardiac function grade(r=0.518,95％CI:0.408～0.613,P＜0.001,r=0.737,95％CI:0.666～0.794,P＜0.001).The poor prognosis rate of patients was 17.93％.Compared with the good prognosis group,the serum CT-1 and ANGPTL3 levels of the poor progno-sis group were increased(P＜0.05).Logistic regression model analysis showed that smoking,diabetes,lesion vessel number ≥3,irregular medication outside the hospital,serum CT-1 and ANGPTL3 levels were the influencing factors of poor prognosis in patients with CHD complicated with HF(P＜0.05).ROC curve analysis showed that the sensitivity and area under the curve(AUC)of combined serum CT-1 and ANGPTL3 levels for predicting poor prognosis of patients with CHD complicated with HF were higher than those of either marker alone,while the specificity was basically similar to that of sin-gle-marker prediction.Conclusion Serum CT-1 and ANGPTL3 levels are abnormally elevated in patients with CHD complicated with HF after PCI,and are closely related to the cardiac function and prognosis.

Objective To reveal the detailed histological characteristics of pyramidal neuron cell bodies and their axonal projections along the corticospinal tract in the primary motor cortex(M1)of the brain,by using the biotinylated dextran amine(BDA)neural tracing technique combined with panoramic and local microscopic imaging technologies.Methods A total of 100 nL of 10％BDA(10,000 molecular weight)was injected into M1 region using stereotaxic system.The distribution of BDA labeling along the corticospinal tract was continuously tracked with panoramic tissue scanning analysis system.Detailed observations of the histological characteristics of BDA labeling were carried out with laser confocal microscope.Results It is more convenient to observe the overall distribution of BDA neural labeling by using the panoramic tissue scanning analysis system.Around the injection site in M1,the BDA labeling was shown in the somas of pyramidal neurons in layer V.In the M1 region corresponding to the contralateral site of the injection site and ipsilateral primary sensory cortex,BDA showed predominantly the anterograde labeled nerve fibers accompanied by a few retrograde labeled neurons.Besides,BDA labeled nerve fibers-including bundles and terminals-projecting to regions such as the ipsilateral striatum,thalamus,internal capsule,cerebral peduncle,and pons,and further reaching the contralateral spinal cord via the brainstem pyramidal decussation.Confocal microscopy and its 3D reconstruction system facilitated detailed analysis of the local microscopic features of BDA labeling,revealing retrograde labeled neuron cell bodies,dendrites and their spines,as well as anterograde labeled nerve fibers and their terminals.Conclusions These findings demonstrated that the integration of traditional BDA neural tracing with panoramic tissue scanning analysis and confocal microscopy provided an effective approach to the observation and analysis of long-projection neural circuits from panoramic to local perspectives,with broad application prospects.

Objective:To study the application of 3D printing technology in multi-center fenestrated/branched endovascular repair (F/B-EVAR) for endovascular repair of abdominal aortic diseases.Methods:From Feb 2018 to Mar 2023, The clinical and followup data of 316 cases of abdominal aortic lesions undergoing repair with F/B-EVAR at 69 medical centers nationwide using 3D printing technology to guide physician-modified stent graft were retrospectively analyzed.Results:The mean follow-up time of the patients was 23 months (2-60 months), and 24 cases were lost to follow up, the follow-up rate was 92.4% (292/316), the mean postoperative hospitalization time was (8.2±4.9) days. A total of 944 main abdominal branch arteries were reconstructed. Intraoperative reconstruction of 11 branches failed, with a success rate of 98.8% (933/944). Within 30 days after surgery, 8 patients died (2.5%), and 6 patients died during follow-up, a total of 14 patients died (4.4%). There were 11 cases (3.5%) of spinal cord ischemia and no patient suffered from permanent paraplegia. There were 19 patients (6.0%) with postoperative renal function injury. Internal leakage was found in 26 patients, and the rate of internal leakage was 8.2%.Conclusion:3D printing technology can accurately locate the location of branch arteries, simplifing the surgical process, shortening the learning curve , and improving clinical efficacy.

This article reported a patient who initially presented with insomnia complaints and was subsequently diagnosed with severe obstructive sleep apnea （OSA） on polysomnography （PSG）. The patient tried continuous positive airway pressure （CPAP）but gave up because wear the ventilator made it more difficult to fall asleep. Then the patient only received group cognitive behavioral therapy for insomnia （CBT-I）， which not only alleviated insomnia severity but also promoted severe OSA into mild status. Such case suggested that， firstly， due to the high comorbidity of insomnia and OSA， evaluation of OSA should be considered a part worth enough attention of the clinical diagnosis and treatment of insomnia patients. Secondly， by relieving insomnia， CBT-I can alleviate both nocturnal apnea and daytime somnolence in patients with comorbid insomnia and sleep apnoea （COMISA）， so the application of CBT-I should be emphasized in the treatment of such patients. ［Funded by the Central Government-guided Local Science and Technology Development Fund Project （number， 2022ZY0028）］