The evolution of hepatocellular carcinoma (HCC) is a compound process which involves many kinds of genes and transductional pathways. The expression of the peptidyl-proplyl-isomerase PIN1 gene, the mutation in exon 3 of beta-catenin and its correspondent abnormal expression and their roles in the hepatocellular carcinogeneisis were investigated. Among 29 pair cases of HCC and non-carcinoma tissues, the expression of PIN1 gene was detected by immunochemical staining. Mutations in exon 3 of beta-catenin gene and differential expression of beta-catenin gene were investigated by the methods of PCR-SSCP, direct sequencing and immunohistochemical technique as well. The results indicated: (1) 44.8% (13/29) cases of HCC presented higher level of PIN1 gene expression than non-cancerous tissues (chi2=32.63, P<0.05), especially in cytoplasm and nucleus, while there was lower level of PIN1 expression in non-cancerous tissues; (2) 58.6% (17/29) HCC tissues showed beta-catenin protein accumulation in cytoplasm and nucleus. 46.2% (6/13) HCC tissues indicated beta-catenin protein accumulation with higher level of PIN1 expression, while 53.8% (7/13) HCC tissues indicated beta-catenin protein accumulation with lower level or trace of PIN1 expression (chi2=0.00, P>0.05); (3) 24.1% (7/29) of primary tumor lesions carried gene mutations in exon 3 of beta-catenin, and accompanied by beta-catenin protein accumulation. There was no mutation in non-cancerous tissues. All the mutation presented in tissues with low level of PIN1 expression. There was no mutation of beta-catenin gene in tissues with high PIN1 expression level (chi2=58.12, P<0.05). So it was postulated that the increase of PIN1 gene expression could promote hepatocellular carcinogenesis via a way different from beta-catenin gene mutation.

Objective:To explore the application effect of medical science competition in nursing interns whocontribute to "healthy China" , and to improve their health education awareness, ability, method and self-confidence.Methods:A total of 205 nursing interns who worked in Union Hospital of Tongji Medical College of Huazhong University of Science and Technology from 2019 to 2020 were selected as the research objects. They were divided into the control group (105 cases) and the experimental group (100 cases) according to whether they participated in the medical science competition. The control group learned the form and method of health education in clinical rotation according to the traditional practice teaching plan. The experimental group volunteered to participate in the medical science competition, which required the dissemination of health knowledge through various forms. Before and after the competition, the health education ability assessment scale was used for comparison.Results:Before the medical science competition, there was no significant difference in the total score of assessment, planning, implementation, evaluation and health education between the control group and the experimental group ( t values were 0.765 - 1.749, all P>0.05). After the medical science competition, the total scores of assessment, planning, implementation, evaluation and health education ability of nursing interns in the experimental group were (24.38 ± 4.72), (17.98 ± 3.98), (25.16 ± 5.36), (12.57 ± 2.96) and (80.09 ± 15.65) respectively, while those in the control group were (22.45 ± 6.29), (16.61 ± 4.77), (23.04 ± 6.55), (11.31 ± 3.46) and (73.41 ± 19.69).The differences between the two groups were statistically significant ( t values were 2.226 - 2.795, all P<0.05). Conclusions:The medical science competition can improve the health education ability of assessment, planning, implementation, evaluation of nursing interns and contribute to "healthy China" .

The evolution of hepatocellular carcinoma (HCC) is a compound process which involves many kinds of genes and transductional pathways. The expression of the peptidyl-proplyl- isomerase PIN1 gene, the mutation in exon 3 of β-catenin and its correspondent abnormal expression and their roles in the hepatocellular carcinogeneisis were investigated. Among 29 pair cases of HCC and non-carcinoma tissues, the expression of PIN1 gene was detected by immunochemical staining. Mutations in exon 3 of β-catenin gene and differential expression of β-catenin gene were investigated by the methods of PCR-SSCP, direct sequencing and immunohistochemical technique as well. The results indicated: (1) 44.8% (13/29) cases of HCC presented higher level of PIN1 gene expression than non-cancerous tissues (x2 =32.63, P＜0.05), especially in cytoplasm and nucleus, while there was lower level of PIN1 expression in non-cancerous tissues; (2) 58.6% (17/29) HCC tissues showed β-catenin protein accumulation in cytoplasm and nucleus. 46.2% (6/13) HCC tissues indicated β-catenin protein accumulation with higher level of PIN1 expression, while 53.8% (7/13) HCC tissues indicated β-catenin protein accumulation with lower level or trace of PIN1 expression (x2 =0.00, P＞0.05); (3) 24.1% (7/29) of primary tumor lesions carried gene mutations in exon 3 of β-catenin, and accompanied by β-catenin protein accumulation. There was no mutation in non-cancerous tissues. All the mutation presented in tissues with low level of PIN1 expression. There was no mutation of β-catenin gene in tissues with high PIN1 expression level (x2=58.12, P＜0.05). So it was postulated that the increase of PIN1 gene expression could promote hepatocellular carcinogenesis via a way different from β- catenin gene mutation.

Objective To investigate the effects of group identification on death anxiety among military medical students and the chain mediating effects of self-esteem and collective self-esteem.Methods Cluster sampling was conducted to survey the students in a military medical university in July,2021,and finally survey data from 360 participants were collected through WeChat Mini Program,Questionnaire Star.The Questionnaires included Templer Death Anxiety Scale(T-DAS),Organizational Identification Questionnaire,Rosenberg Self-Esteem Scale-Revised(RSES-R),Collective Self-Esteem Scale and a self-designed general information questionnaire.Results ①The score of death anxiety in the participants ranged from 1.00 to 4.33(M=2.60).②The scores of group identification,self-esteem and collective self-esteem were negatively correlated with death anxiety(r=-0.56～-0.21,P＜0.01).Significantly positive correlations were observed in any 2 scores among the above 3 scores(r=0.42～0.68,P＜0.01).③ Group identification significantly negatively predicted death anxiety(b=-0.21,SE=0.02,P＜0.001).④ There were 3 mediation effects between group identification and death anxiety:group identification→self-esteem→death anxiety,group identification→ collective self-esteem→ death anxiety,group identification→ self-esteem→collective self-esteem→death anxiety,with a total indirect effect of-017.Conclusion Group identification can negatively predict death anxiety among military medical students,and self-esteem and collective self-esteem play a chain mediating role between them.

OBJECTIVETo establish two double-antigen sandwich ELISA systems to detect anti-Afmp1cr and Afmp2cr antibodies of Aspergillus fumigatus.

METHODSRecombinant 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.

RESULTSThe 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.

CONCLUSIONSTwo 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.

Animals ; Antibodies, Fungal ; isolation & purification ; Antigens, Fungal ; Aspergillosis ; diagnosis ; Aspergillus fumigatus ; Cross Reactions ; Enzyme-Linked Immunosorbent Assay ; Humans ; Pichia ; Rabbits ; Recombinant Proteins ; Sensitivity and Specificity

ObjectiveTo evaluate the supportive policies for caregivers issued in China, and analyze the structures and contents. MethodsThe national supportive policies for family caregivers in China within the past decade were text-mined using ROSTCM 6.0. Secondary indicators were established according to the Policy Modeling Consistency (PMC) index model combining with World Health Organization six blocks of health services. ResultsThe average PMC score was 7.38. There were seven good policies and three accepted policies. The policies covered well in publicity, recipients and functions, and needed to improve in incentive and restraint. ConclusionChina's policies have played a positive role in supporting family caregivers. It is needed to reduce the burden on family caregivers and improve their welfare.

Objective To establish two double-antigen sandwich ELISA systems to detect anti-Afmp1cr and Afmp2cr antibodies of Aspergillus fumigatus. Methods 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. Results 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. Conclusion 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.

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.

Objective To establish two double-antigen sandwich ELISA systems to detect anti-Afmp1cr and Afmp2cr antibodies of Aspergillus fumigatus. Methods 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. Results 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. Conclusion 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.

Objective To construct a universal influenza mRNA vaccine and evaluate its immunogenicity.Methods The antigen sequence of hemagglutinin(HA),nucleoprotein(NP)and matrix protein 2 ectodomain(M2e)in influenza A/California/04/2009 was optimized.HA,NP and 3 tandem M2e(3M2e)were cloned into pcDNA3.1 vector,respectively.Then the mRNAs were synthesized by linearization,in vitro transcription,enzymatic capping and enzymatic tailing,and named as mRNA-HA,mRNA-NP and mRNA-3M2e,respectively.The protein expression of the 3 kinds of mRNAs in 293T cells was detected by immunofluorescence assay.Comb-mRNA vaccine was prepared by enveloped mRNA-HA,mRNA-NP and mRNA-3M2e with lipid nanoparticles,respectively,and the particle size and potential were identified.Twenty-eight 6-week-old female BALB/c mice(18～22 g)were randomly divided into LNP group(n=14)and Comb-mRNA group(n=14).Hemagglutination inhibition(HI)method and microneutralization(MN)test were used to evaluate the serum antibody titer induced by Comb-mRNA vaccines.The mice were infected by 5LD50 wild-type H1 N1 influenza virus to evaluate the protective efficacy.Results The mRNA-HA,mRNA-NP and mRNA-3M2e were successfully constructed,and the 3 mRNAs could be expressed in 293T cells.The average size of mRNA encapsulated by lipid nanoparticles was 119.53±6.5 nm,and the average potential was-8.23±1.3 mV.The geometric mean titer(GMT)of HI and MN in the Comb-mRNA group were 179.6 and 201.6,compared with the LNP group.The ratio of IFN-γ+CD4+/CD8+Tcells was increased.The Comb-mRNA group could provide protection against 5LD50 wild type influenza H1 N1 virus after 2 weeks of booster immunization.Conclusion Comb-mRNA,an influenza vaccine candidate,can induce immune responses and protect mice from influenza virus challenge.