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Microbiology and Immunology is one of core basic curriculums for medical college undergraduates.Combining with characteristics of this course,our teaching comprehension and background calling for curriculum ideological and political education,we consider blending traditional sinology classic"Thirty-Six Stratagems"in teaching of this curriculum.Meanwhile,its practical signif-icance on many aspects will be elucidated.This paper will provide useful reference for blending other sinology classics into ideological and political education of specialized curriculums.
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Objective:To explore a method to obtain more mouse bone marrow cells more quickly.Methods:Take both lower limbs of the same mouse,the bone marrow cells were obtained by either the tube-nesting EP method or the traditional syringe irrigation method,recorded the time and quantity of bone marrow cells,to analyze the proportion of viable cells by flow cytometry,and to detect the proliferation of these bone marrow cells used CCK-8 method.Results:Compared with the traditional syringe irrigation method,the time of obtaining mouse bone marrow cells by tube-nesting EP method was shorter(P<0.000 1).Compared with the traditional syringe irrigation method,the number of bone marrow cells obtained by tube-nesting EP method was more(P<0.05).The proportion of live cells and proliferation ability of bone marrow cells obtained by the two methods were identical.Conclusion:Tube-nesting EP method is a new faster method to get more mouse bone marrow cells.
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Objective:To identify the morphological alterations in the Golgi apparatus of skin fibroblasts in spinocerebellar ataxia type 3 (SCA3) patients.Methods:In this study, 3 SCA3 patients and 3 healthy volunteers were obtained in the First Affiliated Hospital of Zhengzhou University from 2016 to 2020. The cytosine, adenine, and guanine repeats of 3 SCA3 patients were 14/76, 20/80 and 21/82, respectively. Tissue mass culture was used to amplify skin fibroblasts derived from SCA3 patients and healthy volunteers. Cell viability and apoptosis were detected using cell counting kit-8 assay and flow cytometry, respectively. Western blotting and immunofluorescence assay were used to detect the protein expression of ataxin-3, Golgi reassembly stacking protein 2 (GORASP2), and Golgi matrix protein 130 (GM130) in the skin fibroblasts. The morphology of the Golgi apparatus in skin fibroblasts was detected using transmission electron microscopy.Results:Tissue culture of skin fibroblasts of both SCA3 patients and healthy volunteers was successfully established. The patient-derived dermal fibroblasts expressing mutant ataxin-3 protein exhibited reduced cell viability ( t=5.06,P=0.007), increased apoptosis ( t=3.77, P=0.020), fragmentation of the Golgi apparatus, increased expression of GM130 ( t=5.23, P=0.006), and decreased expression of GORASP2 ( t=4.35, P=0.012). Transmission electron microscopy revealed that the Golgi apparatus was disorganized in skin fibroblasts. Conclusion:Fragmentation of the Golgi apparatus occurs in the skin fibroblasts of SCA3 patients, and abnormal morphology and structure of the Golgi apparatus may be involved in the pathogenesis of SCA3.
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@#Objective To investigate differential expression genes （DEGs） between Alzheimer’s disease （AD） and normal controls by bioinformatics analysis.Methods The microarray dataset GSE5281 was download from GEO database，which included brain tissue in AD and normal controls. The DEGs were obtained by R project.Analysis of DEGs based in DAVID database was used to obtain gene ontology（GO）and kyoto encyclopedia of genes and genomes （KEGG） pathway.The protein protein interaction network （PPI） was established using STRING database to identify hub genes. and core genes.Moreover，the existing drugs target to these core genes were screen to explore the therapeutic effect for AD.Results A total 863 DEGs were obtained，of which 246 genes were up-regulated and 617 genes were down-regulated in AD group.GO showed that DEGs were mainly involved in ATP binding，and KEGG pathway involved several neurodegenerative diseases including Parkinson’s disease and prion disease，long-term potentiation and axon guidance.5 core genes（PSMA7，PSMA3，PSMB7，PSMC5 and PSMC3） and 31 hub genes including 23 up regulated genes and 8 down-regulated genes were obtained by PPI analysis.Several existing drugs have targeted to core genes. Some common differential expression genes were obtained by paired comparison of 3 groups of gene microarrays.Conclusion Bioinformatics analysis based on GEO database showed that there were DEGs between Alzheimer’s disease（AD）and normal controls，and 8 existing drugs were identified.
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Objective To investigate the real-time regulatory effects of IFN-γ, programed death ligand 2(PDL2) and janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway on the adherence, proliferation and migration of human placenta-derived mesenchymal stem cells(hPMSCs) based on a finding that IFN-γ could enhance the expression of PDL2 in hPMSCs through JAK/STAT signaling pathway.Methods hPMSCs were isolated by using enzyme digestion method and then co-cultured with IFN-γ, anti-PDL2 monoclonal antibody (anti-PDL2 McAb) and JAK inhibitor, respectively.Real-time cell analysis (RTCA) was used to detect the dynamic changes in the adherence, proliferation and migration of hPMSCs following various interventions.Results IFN-γ remarkably suppressed hPMSCs proliferation during the period from 40 hours to 80 hours after intervention and also inhibited the non-targeted migration of hPMSCs.However, hPMSCs adherence was not affected by IFN-γ.Co-culturing hPMSCs with anti-PDL2 McAb significantly enhanced hPMSCs adhesion and inhibited their non-targeted migration, but had no significant effect on hPMSCs proliferation.Furthermore, the proliferation of hPMSCs co-cultured with IFN-γ and anti-PDL2 McAb was significantly inhibited as compared with that of anti-PDL2McAb treatment group.The adhesion, migration and proliferation of hPMSCs were significantly inhibited after co-culturing them with JAK inhibitor.Conclusion IFN-γ can remarkably suppress the proliferation and migration of hPMSCs.PDL2 can enhance the migration and inhibit the adhesion of hPMSCs.JAK/STAT signaling pathway is involved in regulating the adhesion, migration and proliferation of hPMSCs.
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<p><b>OBJECTIVE</b>To establish two double-antigen sandwich ELISA systems to detect anti-Afmp1cr and Afmp2cr antibodies of Aspergillus fumigatus.</p><p><b>METHODS</b>Recombinant Afmp1cr and Afmp2cr proteins of A.fumigatus expressed in Pichia pastoris were obtained. Double-antigen sandwich ELISA systems for detecting specific anti-Afmp1cr and anti-Afmp2cr antibodies were developed after chessboard titrating to determine the appropriate concentrations of the recombinant proteins and HRP-labeled proteins. The sensitivity of the assay was evaluated using serum samples of rabbits immunized with Afmp1cr and Afmp2cr. The specificity of the assay was evaluated by detecting serum samples from healthy donors and patients with other pathogenic fungal and baterial infections. The performance of the two ELISA kits was furthered evaluated using serum samples from patients with suspected Aspergillus infection.</p><p><b>RESULTS</b>The established ELISA kits were capable of detecting anti-Afmp1cr and anti-Afmp2cr antibodies in immunized rabbit serum at the maximum dilutions of 800 and 3200, respectively. No cross-reactivity was observed in detecting serum from patients with other pathogenic fungal or bactetial infections. Both of the two kits yielded positive results in sera from two established Aspergillus-infected cases and a suspected case.</p><p><b>CONCLUSIONS</b>Two antibody-capture ELISA kits were developed for the laboratory diagnosis of A.fumigatus infection and can be potentially useful in the clinical diagnosis of Aspergillosis infections.</p>