A retrospective cohort study of Epstein-Barr virus infection status and systemic lupus erythematosus.

OBJECTIVES: Systemic lupus erythematosus (SLE) and the Epstein-Barr virus (EBV) are very closely related. This study estimated the impact of EBV infection status on clinical manifestations and disease remission in patients with SLE. METHOD: A retrospective study was performed using electronic health records of patients with SLE. The SLE disease activity index (SLEDAI-2 K) was used to assess disease activity. VCAIgM or EAIgM positive or EBVDNA copies ≥ 50 IU/mL were defined as lytic infection group, EBNA-IgG or VCAIgG-positive and who were negative for both VCAIgM and EAIgM with EBVDNA copies < 50 IU/mL were defined as the latent infection group. The endpoint (disease remission) was defined as a decrease in SLEDAI-2 K score of ≥ 1 grade or ≥ 4 points from baseline. The association between EBV infection status and disease remission was assessed using propensity score weighting and multivariable Cox regression models. RESULTS: There were 75 patients with SLE in the EBV lytic infection group and 142 patients in the latent infection group. The SLEDAI-2 K score was higher in the lytic infection group (10.00 (6.25, 16.00) vs. 8.00 (5.00, 10.00), Z = 3.96, P < 0.001). There was a significant difference in the effect of EBV lytic infection on disease remission compared to latent infection (HR 0.30, 95% CI 0.19-0.49, P < 0.001). CONCLUSIONS: Patients with SLE with lytic EBV infection have higher disease activity and take longer to achieve remission. Our study furthers our understanding of the relationship between SLE and EBV infection and may inform better treatment practices in the future.

The relationship between quality of life and health promotion behavior in patients with type B aortic dissection: a cross-sectional study.

BACKGROUND: To understand the current situation of health promotion behavior and quality of life among aortic dissection survivors and the correlation between them. METHODS: Sociodemographic characteristics were collected. T-test and variance analysis were applied for univariate analysis. Quality of life was measured using the SF-36 Questionnaire, and health-promoting behaviors were measured using the aortic dissection health promotion behavior questionnaire. The association between type B aortic dissection survivors' health promotion behavior and health status questionnaire (SF-36) scores was determined through Pearson's correlation coefficients. This association was analyzed through multivariable regression analysis. RESULTS: A total of 131 type B aortic dissection survivors were evaluated through the self-developed aortic dissection patient health promotion behavior scale and health status questionnaire (SF-36). Results showed that the health promotion behavior of Stanford B aortic dissection survivors (85.05 ± 11.28) correlated with their Mental Component Summary (MCS) (55.23 ± 30.72; r = 0.359, P < 0.01). The model showed 39.00% variance shared between behavior motivation and MCS (R2 = 0.390, F = 13.189, P < 0.01). CONCLUSION: Type B aortic dissection survivors in Zunyi, China had a lower quality of life. Medical staff can formulate intervention measures from behavioral motivation to improve the quality of life of aortic dissection survivors.

Altered Cellular Immunity and Differentially Expressed Immune-Related Genes in Patients With Systemic Sclerosis-Associated Pulmonary Arterial Hypertension.

Systemic sclerosis (SSc) is the most common connective tissue disease causing pulmonary hypertension (PAH). However, the cause and potential immune molecular events associated with PAH are still unclear. Therefore, it is particularly essential to analyze the changes in SSc-PAH-related immune cells and their immune-related genes. Three microarray datasets (GSE22356, GSE33463, and GSE19617) were obtained by the Gene Expression Omnibus (GEO). Compared with SSc, we found neutrophils have a statistically higher abundance, while T-cell CD4 naive and T-cell CD4 memory resting have a statistically lower abundance in peripheral blood mononuclear cells (PBMCs). Moreover, the results of Gene Set Enrichment Analysis (GSEA) showed there is a differential enrichment of multiple pathways between SSc and SSc-PAH. By combining differentiated expressed genes (DEGs) and immune-related genes (IRGs), fifteen IRGs were selected. In addition, we also analyzed the first five rich Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways and the most abundant Gene Ontology (GO)-molecular functional terms. Furthermore, interleukin-7 receptor (IL-7R), tyrosine-protein kinase (LCK), histone deacetylase 1 (HDAC1), and epidermal growth factor receptor (EGFR) genes were identified as hub genes via protein-protein interaction (PPI) network analysis. The Comparative Toxic Genomics Database (CTD) analysis result showed that LCK, HDAC1, and EGFR have a higher score with SSc. Coexpression network analysis confirmed that IL-7R, LCK, and HDAC1 are key genes related to immune regulation in SSc without PAH and are involved in T-cell immune regulation. Subsequently, using GSE22356 and GSE33463 as the test sets and GSE19617 as the verification set, it was verified that the mRNA expression levels of the three central genes of SSc-PAH were significantly lower than those of the SSc without PAH samples. Consistent with previous predictions, the expressions of IL-7R, LCK, and HDAC1 are positively correlated with the numbers of T-cell CD4 naive and T-cell CD4 memory, while the expressions of IL-7R and LCK are negatively correlated with the numbers of neutrophils in the peripheral blood. Therefore, this evidence may suggest that these three immune-related genes: IL-7R, LCK, and HDAC1, may be highly related to the immunological changes in SSc-PAH. These three molecules can reduce T cells in SSc-PAH PBMCs through the regulation of T-cell activation, which suggests that these three molecules may be involved in the development of SSc-PAH. Meanwhile, the low expression of IL-7R, LCK, and HDAC1 detected in the peripheral blood of SSc may indicate the possibility of PAH and hopefully become a biomarker for the early detection of SSc-PAH. Finally, 49 target miRNAs of 3 specifically expressed hub genes were obtained, and 49 mRNA-miRNA pairs were identified, which provided directions for our further research.

Comprehensive Analysis to Identify MAGEA3 Expression Correlated With Immune Infiltrates and Lymph Node Metastasis in Gastric Cancer.

Gastric cancer (GC) is an aggressive malignant tumor and causes a significant number of deaths every year. With the coming of the age of cancer immunotherapy, search for a new target in gastric cancer may benefit more advanced patients. Melanoma-associated antigen-A3 (MAGEA3), one of the members of the cancer-testis antigen (CTA) family, was considered an important part of cancer immunotherapy. We evaluate the potential role of MAGEA3 in GC through the TCGA database. The result revealed that MAGEA3 is upregulated in GC and linked to poor OS and lymph node metastasis. MAGEA3 was also correlated with immune checkpoints, TMB, and affected the tumor immune microenvironment and the prognosis of GC through CIBERSORT, TIMER, and Kaplan-Meier plotter database analysis. In addition, GSEA-identified MAGEA3 is involved in the immune regulation of GC. Moreover, the protein-protein interaction (PPI) networks of MAGEA3 were constructed through STRING database and MAGEA3-correlated miRNAs were screened based on the joint analysis of multiple databases. In terms of experimental verification, we constructed pET21a (+)/MAGEA3 restructuring plasmids and transformed to Escherichia coli Rosetta. MAGEA3 protein was used as an antigen after being expressed and purified and can effectively detect the specific IgG in 93 GC patients' serum specimens with 44.08% sensitivity and 92.54% specificity. Through further analysis, the positive rate of MAGEA3 was related to the stage and transfer number of lymph nodes. These results indicated that MAGEA3 is a novel biomarker and correlated with lymph node metastasis and immune infiltrates in GC, which could be a new target for immunotherapy.

Analysis of lncRNA expression profiles by sequencing reveals that lnc-AL928768.3 and lnc-AC091493.1 are novel biomarkers for disease risk and activity of rheumatoid arthritis.

BACKGROUND: This study aimed to analyze long non-coding RNA (lncRNA) expression profiles in synovium tissue of patients with RA using RNA sequencing, and to further assess the clinical values of dysregulated lncRNAs in RA diagnosis and monitoring. METHODS: Thirty patients with RA who underwent knee arthroscopy and 30 controls with knee trauma who underwent surgery were consecutively enrolled and synovium tissue samples of both groups were obtained during surgery. In the exploration stage, lncRNA and mRNA expression profiles in three RA samples and three control samples were detected by RNA sequencing and bioinformatic analyses were then performed. In the validation stage, quantitative polymerase chain reaction (qPCR) was subsequently used to detect expression of five candidate lncRNAs in 30 patients with RA and 30 control patients. RESULTS: A total of 349 lncRNAs and 1582 mRNAs were upregulated and 806 lncRNAs and 1295 mRNAs were downregulated in patients with RA compared with controls. Enrichment analyses revealed that these dysregulated lncRNAs and mRNAs were mainly involved in regulating immune response, leukocyte migration, complement activation, and B cell receptor signaling pathway. Subsequent qPCR validation discovered that lnc-AL928768.3 (P < 0.001) and lnc-AC091493.1 (P < 0.001) were elevated in patients with RA compared with controls and afford good predictive values for RA risk by receiver operating characteristic (ROC) curve analysis. Additionally, the two lncRNAs were positively associated with C-reactive protein level and disease activity score in 28 joints (ESR) (all P < 0.05). CONCLUSION: Analysis of lncRNA expression profiles by sequencing reveals that lnc-AL928768.3 and lnc-AC091493.1 are novel biomarkers for RA risk and activity.

MiR-5571-3p and miR-135b-5p, derived from analyses of microRNA profile sequencing, correlate with increased disease risk and activity of rheumatoid arthritis.

OBJECTIVES: This study aimed to investigate microRNA (miRNA) expression profiles in synovium tissues of rheumatoid arthritis (RA) patients by RNA sequencing and to evaluate the values of dysregulated miRNAs in RA diagnosis and monitoring. METHODS: Thirty RA patients who underwent knee arthroscopy and 30 controls with knee trauma who underwent surgery were consecutively recruited, and synovium tissue samples of both groups were obtained during surgeries. In the exploration part, miRNA and mRNA expression profiles of 3 RA samples and 3 control samples were detected using RNA sequencing then followed by bioinformatic analyses. In the validation part, 5 candidate miRNA levels were detected by quantitative polymerase chain reaction (qPCR) in 30 RA patients and 30 control patients. RESULTS: In the exploration part, 78 miRNAs and 1582 mRNAs were upregulated while 40 miRNAs and 1295 mRNAs were downregulated in synovium tissue samples of RA patients compared with those of controls. Furthermore, enrichment analyses revealed that these dysregulated miRNAs and mRNAs were mainly implicated in immune activities and inflammatory diseases such as leukocyte migration, complement activation, and RA. In the validation part, qPCR assay revealed that miR-5571-3p and miR-135b-5p expressions were increased in RA patients compared with those in controls and disclosed good predictive values for RA risk with high area under the curves (AUCs). Besides, both miR-5571-3p and miR-135b-5p levels were positively correlated with disease activity and inflammation level of RA. CONCLUSIONS: Analyses of miRNA expression profiles by sequencing indicate that miR-5571-3p and miR-135b-5p correlate with increased RA risk and activity.

The Possible Effect of B-Cell Epitopes of Epstein-Barr Virus Early Antigen, Membrane Antigen, Latent Membrane Protein-1, and -2A on Systemic Lupus Erythematosus.

This study was aimed to evaluate the role of B-cell epitopes of Epstein-Barr virus (EBV) Early antigen protein D (EA), envelope glycoprotein GP340/membrane antigen (MA), latent membrane protein (LMP)-1, and LMP-2A in systemic lupus erythematosus (SLE). B-cell epitopes were predicted by analyzing secondary structure, transmembrane domains, surface properties, and homological comparison. 60 female mice were randomized equally into 12 groups: 1-10 groups were immunized by epitope peptides (EPs) 1-10, respectively, while 11 and 12 groups were PBS and Keyhole limpet hemocyanin (KLH) control groups. Immunoglobulin G (IgG) and autoantibody to nuclear antigen (ANA) concentrations in mice serum were determined at week 8. Indirect levels of EP1-10 were further detected by enzyme-linked immuno sorbent assay (ELISA) in 119 SLE patients and 64 age- and gender-matched health controls (HCs). 10 probable EBV EA, MA, LMP-1, and LMP-2A B-cell epitopes related to SLE self-antigens were predicted and corresponding EP1-10 were synthesized. IgG concentrations at week 8 were increased in EP1-10 and KLH groups compared with PBS group in mice; while ANA levels were elevated in only EP1-4, EP6-7, and EP10 groups compared to KLH group by ELISA, and ANA-positive rates were increased in only EP1, EP2, EP4, EP6, and EP10 groups by indirect immunofluorescence assay. EP1-4, EP6, and EP10 indirect levels were increased in SLE patients than HCs, while EP1, EP3, EP6, and EP9 were correlated with SLE disease activity index score. In conclusion, EBV EA, MA, LMP-1, and LMP-2A B-cell EPs increased SLE-related autoantibodies in mice, and their indirect levels might be served as potential biomarkers for SLE diagnosis and disease severity.

[Expression of HPV16 E7 protein and preparation of its polyclonal antibody].

OBJECTIVE: To prepare a prokaryotic expression vector carrying E7 protein of human papillomavirus (HPV) type 16 and a polyclonal antibody against it. METHODS: The HPV16 E7 gene was amplified by PCR from the tissue samples of cervical cancer and cloned into the pET21a(+) prokaryotic expression vector. The constructed pET21a(+)/HPV16 E7 recombinant plasmid was transformed into E.coli BL21(DE3) and induced by isopropyl beta-D-1-thiogalactopyranoside (IPTG). The recombinant protein of HPV16 E7 was purified by Ni-NTA affinity chromatography and confirmed by SDS-PAGE and Western blot analysis. To prepare the polyclonal antibody, the rabbits were immunized with the purified recombinant protein HPV16 E7. The titers of specific antibodies were detected by ELISA. The specificity activity was further detected by Western blotting and immunofluorescence analysis. RESULTS: HPV16 E7 recombinant protein was expressed and purified in the prokaryotic expression system. The polyclonal antibodies were produced in the rabbits immunized with the recombinant protein. The titer of specific antibodies was up to 1:30 000. Western blotting and immunofluorescence analysis indicated that the polyclonal antibody could specifically recognize the HPV16 E7 protein. CONCLUSION: HPV16 E7 recombinant protein was successfully expressed and its polyclonal antibody was prepared.