This study investigated the full-genome molecular characteristics of a rotavirus vaccine-derived strain,G1P[8]geno-type A group rotavirus RVA/Human-wt/CHN/HN1140/2021/G1P[8](referred to as HN1140).The gene fragments of the HN1140 strain were amplified with reverse transcription-polymerase chain reaction(RT-PCR)combined with whole-genome primers to obtain the full genome sequence.Genotyping was performed with the online genotyping tool RotaC 2.0,and similarity and genetic evolution analyses for each gene segment were conducted in DNAstar5.1 and MEGA11.0 software.The genotype of the HN1140 strain was deter-mined to be G1-P[8]-I2-R2-C2-M2-A3-N2-T6-E2-H3.Phylogenetic analysis demonstrated that all 11 genomic segments clus-tered closely with the RotaTeq vaccine strains,sharing 99.7%-100%nucleotide sequence similarity.Notably,VP1,VP2,VP6,and NSP2-NSP5 segments showed 100%nucleotide identity with RotaTeq strains.Comparative genomic analysis identified 13 nucleotide and 8 amino acid substitutions between HN1140 and RotaTeq strains,localized within the VP7,VP4,VP1,VP2,VP3,and NSP1 segments.The HN1140 strain exhibited the genotype G1-P[8]-A3-T6-H3,which was consistent with the typical profile of a vaccine-derived reassortant.This strain demonstrated high genetic similarity to RotaTeq vaccine strains,with nucleotide sequence identity ranging from 99.7%to 100%.These findings suggested that HN1140 evolved from RotaTeq vaccine strains through genetic reassortment.

This study investigated the full-genome molecular characteristics of a rotavirus vaccine-derived strain,G1P[8]geno-type A group rotavirus RVA/Human-wt/CHN/HN1140/2021/G1P[8](referred to as HN1140).The gene fragments of the HN1140 strain were amplified with reverse transcription-polymerase chain reaction(RT-PCR)combined with whole-genome primers to obtain the full genome sequence.Genotyping was performed with the online genotyping tool RotaC 2.0,and similarity and genetic evolution analyses for each gene segment were conducted in DNAstar5.1 and MEGA11.0 software.The genotype of the HN1140 strain was deter-mined to be G1-P[8]-I2-R2-C2-M2-A3-N2-T6-E2-H3.Phylogenetic analysis demonstrated that all 11 genomic segments clus-tered closely with the RotaTeq vaccine strains,sharing 99.7%-100%nucleotide sequence similarity.Notably,VP1,VP2,VP6,and NSP2-NSP5 segments showed 100%nucleotide identity with RotaTeq strains.Comparative genomic analysis identified 13 nucleotide and 8 amino acid substitutions between HN1140 and RotaTeq strains,localized within the VP7,VP4,VP1,VP2,VP3,and NSP1 segments.The HN1140 strain exhibited the genotype G1-P[8]-A3-T6-H3,which was consistent with the typical profile of a vaccine-derived reassortant.This strain demonstrated high genetic similarity to RotaTeq vaccine strains,with nucleotide sequence identity ranging from 99.7%to 100%.These findings suggested that HN1140 evolved from RotaTeq vaccine strains through genetic reassortment.

BACKGROUND:Studies have found that glucagon-like peptide-1 and its analogues have a significant neuroprotective effect,and some drugs have been applied to the clinical stage Ⅲ study of Alzheimer's disease.However,the mechanism of its neuroprotective effect is still unclear,which needs to be further explored and clarified. OBJECTIVE:To screen out the genes related to the pathogenesis of Alzheimer's disease and the related targets of semaglutide for the treatment of Alzheimer's disease based on bioinformatics and network pharmacology analyses,to identify the potential target genes by comprehensive analysis of the two and to verify them at the cellular level. METHODS:Using DisGeNET database,differentially expressed genes between Alzheimer's disease patients and healthy population were screened out.The chemical structure formula and two-dimensional structure diagram of semaglutide were obtained using PubChem online database.GO/KEGG enrichment analysis was performed using DAVID online database.A protein-protein interaction network was constructed by using the STRING database.The HPA database was used to determine the distribution characteristics of the target proteins in various human tissues.Finally,western blot was used to detect relevant protein expression in HT22 cells after semaglutide intervention. RESULTS AND CONCLUSION:With the dataset in DisGeNET database,3 374 differentially expressed genes between Alzheimer's disease patients and healthy people were obtained,and meanwhile,101 target genes of semaglutide potential drugs were obtained.There were 23 intersection genes between them.Ten key genes were identified based on the protein-protein interaction network,which were silent information regulator 1(SIRT1),CASP9,CCND1,CASP1,KEAP1,DLG4,CASP4,GRB2,GRIA1,and EDNRA.The results of GO gene functional annotation analysis of key genes showed that the positive regulatory activity of cysteine endopeptidase,the positive regulation of proteolysis,and the positive regulation of cysteine endopeptidase involved the cytoplasmic part of the apoptotic activity process;AMPA glutamate receptor complex,inflammatory complex,CARD domain binding,cysteine endopeptidase activity,and cysteine endopeptidase activity were involved in the apoptotic process.The results of KEGG signaling pathway analysis indicated that colorectal cancer,non-small cell carcinoma,and endometrial carcinoma were related to immune infiltration,inflammation and autophagic apoptosis.In addition,according to the association ranking of key genes and their distribution in different tissues of HPA online database,SIRT1 was identified as the most significant differential gene.The expression level of SIRT1 protein was significantly down-regulated in HT22 cells after β-amyloid protein 1-42 treatment,but it could be significantly increased after being treated with semaglutide.To conclude,SIRT1 may be a target gene for semaglutide in the treatment of Alzheimer's disease.

Objective To use bioinformatics analysis to identify the key genes and signaling pathways involved in cardiac dysfunction after acute ischemic stroke.Methods The GSE102558 dataset was downloaded from the Gene Expression Omnibus(GEO)database,and genes with values of P<0.05 and|log2FC|>0.6 were identified as being differentially expressed.The MCODE plugin in Cytoscape software performed a functional module analysis of the protein-protein interaction(PPI)network,while the CytoHubba plugin screened for core genes.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were also performed.A middle cerebral artery occlusion model was constructed to verify core gene expression using real-time PCR.Results Among the screened differential genes,385 were upregulated and 354 were downregulated.The top ten core genes were Col1a1,Col1a2,Col3a1,Fbn1,Postn,Col5a1,Mmp3,Eln,Acta2,and Timp3.The GO enrichment mainly involved the extracellular matrix,the collagen fiber tissue,vascular development,and protease binding.KEGG was mainly enriched in protein digestion and absorption,the relaxin pathway,advanced gly-cation end products-receptor for advanced glycation end products,and platelet activation.Real-time PCR verified that Col3a1and Postn expressions decreased in the heart tissue after acute ischemic stroke.Conclusion Col3a1and Postnexpressions may be closely associ-ated with cardiac dysfunction occurrence and development after acute ischemic stroke.

Objective:To establish a rapid quantitative PCR (qPCR) technique for Mycobacterium marinum skin infections, and to analyze its clinical diagnostic efficiency. Methods:DNA was extracted from Mycobacterium marinum colonies and serially diluted (10 -1 to 10 -8). Twelve pairs of previously reported primers and probes, as well as 6 pairs of newly designed primers and probes in this study, were used for qPCR amplification to identify the most sensitive primers and probes for the detection of Mycobacterium marinum. Skin lesion tissues were collected from 72 patients with confirmed Mycobacterium marinum infections (experimental group) and 68 with other mycobacterial infections (control group) at Shandong Provincial Hospital for Skin Diseases & Shandong Provincial Institute of Dermatology and Venereology, Shandong First Medical University & Shandong Academy of Medical Sciences in 2021. These skin tissues were subjected to qPCR amplification, interferon-gamma release assay (IGRA), acid-fast staining, and tissue culture to evaluate the diagnostic efficacy. Results:The newly designed primers and probes targeting the mycobacterial enhanced infection locus 2 (Mel2) demonstrated the highest sensitivity, with a detection limit of 0.86 copies/μl (cycle threshold value = 37) ; the qPCR amplification with the Mel2 primers/probes did not yield positive results when used for the detection of other mycobacteria (including Mycobacterium leprae and Staphylococcus spp) . Among the 72 patients in the experimental group, 44 were positive for qPCR with a sensitivity of 61.1% (95% CI: 49.6% - 71.5%), and 47 were positive for culture with a sensitivity of 65.2% (95% CI: 53.8% - 75.3%) ; all the 68 controls were negative for both qPCR and culture, with their specificities both being 100%. Among 65 patients subjected to IGRA, 31 were positive with a sensitivity of 47.7% (95% CI: 36.0% - 59.6%), while 16 out of 25 controls were negative for IGRA with a specificity of 64.0% (95% CI: 44.5% - 79.8%). Among 58 patients subjected to acid-fast staining, 37 were positive with a sensitivity of 63.8% (95% CI: 50.9% - 74.9%), and 52 out of 66 controls were negative for acid-fast staining with a specificity of 78.8% (95% CI: 67.5% - 86.9%). The combination of qPCR and culture resulted in a sensitivity of 93% and a specificity of 100% for the detection of Mycobacterium marinum. Conclusion:In this study, a highly sensitive qPCR assay was developed for the detection of Mycobacterium marinum, and its combination with culture could further improve the detection sensitivity.

Objective:To express and block the VP7 protein of human group H rotavirus (group H rotavirus, RVH) J19, and to prepare the polyclonal adsorbent polyantibody function of VP7 for identification.Methods:The VP7 protein of human H rotavirus J19 strain was expressed by baculovirus system, and the protein was blocked by affinity chromatography and hydrophobic chromatography. The VP7 polyantiserum was prepared by immunizing the VP7 protein of human H group H rotavirus J19, and the VP7 polyantiserum was identified by ELISA, Western blot (WB) and immunofluorescence.Results:Soluble J19 VP7 protein was obtained. J19 VP7 rabbit polyclonal antibody was prepared. ELISA showed that VP7 polyclonal antibody had a good binding effect, binding to human RVH VP7 protein at 2 048 000-fold. WB showed that the polyclonal antibody could bind to J19 VP7 protein; Immunofluorescence assay showed that VP7 polyclonal antibody could specifically recognize human group H rotavirus J19.Conclusions:The J19 VP7 protein of human H rotavirus was successfully obtained, and the prepared VP7 polyclonal antibody could recognize human H rotavirus, which provided a basis for the detection of human H rotavirus and the study of the structure and function of the viral capsid.

Objective:To express and block the VP7 protein of human group H rotavirus (group H rotavirus, RVH) J19, and to prepare the polyclonal adsorbent polyantibody function of VP7 for identification.Methods:The VP7 protein of human H rotavirus J19 strain was expressed by baculovirus system, and the protein was blocked by affinity chromatography and hydrophobic chromatography. The VP7 polyantiserum was prepared by immunizing the VP7 protein of human H group H rotavirus J19, and the VP7 polyantiserum was identified by ELISA, Western blot (WB) and immunofluorescence.Results:Soluble J19 VP7 protein was obtained. J19 VP7 rabbit polyclonal antibody was prepared. ELISA showed that VP7 polyclonal antibody had a good binding effect, binding to human RVH VP7 protein at 2 048 000-fold. WB showed that the polyclonal antibody could bind to J19 VP7 protein; Immunofluorescence assay showed that VP7 polyclonal antibody could specifically recognize human group H rotavirus J19.Conclusions:The J19 VP7 protein of human H rotavirus was successfully obtained, and the prepared VP7 polyclonal antibody could recognize human H rotavirus, which provided a basis for the detection of human H rotavirus and the study of the structure and function of the viral capsid.

Objective:To express and purify VP7 protein of group A rotavirus (RVA) G1P[8]. The VP7 polyclonal antibody was prepared and its function was evaluated.Methods:The G1 VP7 protein was expressed by baculovirus expression system and purified by affinity chromatography. Polyclonal antibody against G1 VP7 was obtained by immunizing rabbits with G1 VP7 protein. The function of the G1 VP7 polyclonal antibody was verified by Western blotting (WB), enzyme-linked immunosorbent assay (ELISA), and immunofluorescence assay.Results:The soluble G1 VP7 protein of human RVA G1P[8] was obtained using the baculovirus expression system and the VP7 protein was mainly in trimer state. The G1 VP7 polyclonal antibody was prepared and displayed relatively high binding titer to G1 VP7 protein by ELISA. The VP7 polyclonal antibodies could recognize multiple G-type RVAs by WB and ELISA. Immunofluorescence assay further demonstrated that G1 VP7 polyclonal antibody can bind to different RVAs, including Wa (genotype G1P[8]), DS-1(genotype G2P[4]), SA11 (genotype G3P[2]), and human G9P[8] RV strains. In addition, double sandwich ELISA showed that VP7 polyclonal antibody could be used to detect rotavirus in clinical samples.Conclusions:The soluble G1 VP7 protein was successfully expressed and VP7 antibody was obtained. The G1 VP7 polyclonal antibody could bind to a variety of G-type rotaviruses, which lays a foundation for the establishment of detection method of different G type rotaviruses.

Objective:To investigate the diagnosis and treatment of pilots with patent foramen ovale resulting in encephalomalacia foci and discuss the aeromedical assessment principles.Methods:The clinical manifestations, cranial MRI, cardiac ultrasound and transesophageal echocardiography of a pilot with patent foramen ovale and encephalomalacia foci were retrospectively analyzed, and the relevant literatures were reviewed.Results:The patient was a male fighter pilot, 28 years old, without clinical symptoms and complaints of discomfort. A left temporal pole arachnoid cyst (2.18 cm×1.11 cm) and a left frontal brain softening foci (1.80 cm×3.50 cm×3.30 cm) were detected by cranial MRI during physical examination, and patent foramen ovale was detected by transesophageal echocardiography and foam test after admission. The oval foramen was occluded under local anesthesia. The patient was well-recovered after surgery and without discomfort. The cardiac ultrasound recheck showed normal myocardial echoes in the septum and left ventricular wall, no abnormalities in wall thickness and motion amplitude, and coordinated ventricular wall motion. Strong echogenicity of the blocker was seen in the middle of the atrial septum, and the position of the blocker was normal. The aeromedical assessment conclusion of the patient was waiver to flight.Conclusions:The patent foramen ovale should be screened when pilot is diagnosed as encephalomalacia foci or cerebral infarction. The regular reexamination and follow-up should be conducted for the transcatheter occlusion of patent foramen ovale. Pilots could be waivered when they have no clinical symptoms and signs, no arrhythmias and at proper position in dynamic electrocardiogram and submaximal treadmill exercise test, no residual shunt, valve regurgitation, or occluder erosion in imaging examination, and normal cardiac function, reserve function, and regulatory function, normal pressurized breathing and without arrhythmias in centrifuge run, and the grounding observation period is not less than 6 months.

Objective:To investigate the diagnosis and treatment of pilots with patent foramen ovale resulting in encephalomalacia foci and discuss the aeromedical assessment principles.Methods:The clinical manifestations, cranial MRI, cardiac ultrasound and transesophageal echocardiography of a pilot with patent foramen ovale and encephalomalacia foci were retrospectively analyzed, and the relevant literatures were reviewed.Results:The patient was a male fighter pilot, 28 years old, without clinical symptoms and complaints of discomfort. A left temporal pole arachnoid cyst (2.18 cm×1.11 cm) and a left frontal brain softening foci (1.80 cm×3.50 cm×3.30 cm) were detected by cranial MRI during physical examination, and patent foramen ovale was detected by transesophageal echocardiography and foam test after admission. The oval foramen was occluded under local anesthesia. The patient was well-recovered after surgery and without discomfort. The cardiac ultrasound recheck showed normal myocardial echoes in the septum and left ventricular wall, no abnormalities in wall thickness and motion amplitude, and coordinated ventricular wall motion. Strong echogenicity of the blocker was seen in the middle of the atrial septum, and the position of the blocker was normal. The aeromedical assessment conclusion of the patient was waiver to flight.Conclusions:The patent foramen ovale should be screened when pilot is diagnosed as encephalomalacia foci or cerebral infarction. The regular reexamination and follow-up should be conducted for the transcatheter occlusion of patent foramen ovale. Pilots could be waivered when they have no clinical symptoms and signs, no arrhythmias and at proper position in dynamic electrocardiogram and submaximal treadmill exercise test, no residual shunt, valve regurgitation, or occluder erosion in imaging examination, and normal cardiac function, reserve function, and regulatory function, normal pressurized breathing and without arrhythmias in centrifuge run, and the grounding observation period is not less than 6 months.