The protective effects of azilsartan against oscillatory shear stress-induced endothelial dysfunction and inflammation are mediated by KLF6.

BACKGROUND AND PURPOSE: Atherosclerosis is a common cardiovascular disease with high morbidity and mortality. It is reported to be related to oscillatory shear stress (OSS)-induced endothelial dysfunction and excessive production of inflammatory factors. Azilsartan, a specific antagonist of the angiotensin II receptor, has been approved for the management of hypertensive subjects with diabetes mellitus type II (DMII). The present study will investigate the effects of azilsartan against OSS-induced endothelial dysfunction and inflammation, as well as the underlying mechanism. MATERIALS AND METHODS: Cell viability was detected using an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay were used to determine the expression levels of IL-6, TNF-α, IL-1ß, VCAM-1, and ICAM-1 in human aortic endothelial cells (HAECs). Generation of reactive oxygen species (ROS) was measured using 2'-7'dichlorofluorescin diacetate (DCFH-DA) staining, and the level of reduced glutathione (GSH) was evaluated using a commercial kit. The adhesion of THP-1 monocytes to HAECs was evaluated using calcein-AM staining. The expression level of KLF6 was determined using qRT-PCR and Western blot analysis. RESULTS: According to the result of the MTT assay, 5 and 10 µM azilsartan were considered as the optimized concentrations applied in the present study. The elevated production of IL-6, TNF-α, and IL-1ß, increased levels of ROS, decreased levels of reduced GSH, upregulated VCAM-1, ICAM-1, and E-selectin, and the aggravated adhesion of THP-1 cells to HAECs induced by OSS were all reversed by the introduction of azilsartan. The downregulation of KLF6 induced by OSS was significantly reversed by azilsartan. By knocking down the expression of KLF6, the suppressed adhesion of THP-1 cells to the HAECs, and the downregulation of VCAM-1 and ICAM-1 induced by azilsartan in OSS-stimulated HAECs were greatly reversed. CONCLUSION: The protective effects of azilsartan against OSS-induced endothelial dysfunction and inflammation might be mediated by KLF6.

CTLA-4 gene polymorphisms associate with efficacy of postoperative radioiodine-131 for differentiated thyroid carcinoma.

AIM: To investigate the association of CTLA-4 polymorphisms with efficacy of postoperative radioiodine-131 (I-131) treatment for differentiated thyroid carcinoma (DTC). METHODS: A total of 324 DTC patients and 350 healthy individuals were enrolled in our study. Patients received I-131 remnant ablation following surgical resection. Based on the treatment efficacy, patients were divided into the effective (n = 183) and ineffective groups (n = 141). CTLA-4 polymorphisms (+49A>G, CT60A>G and -318C>T) were genotyped by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: AG + AA genotype distribution and A allele frequency of +49A>G and CT60A>G polymorphisms were higher in the effective group than the ineffective group. CONCLUSION: +49A>G and CT60A>G polymorphisms were associated with the efficacy of postoperative I-131 treatment for DTC; and they might be bioindicators related to the prognosis of I-131 treatment.

Switching and emergence of CTL epitopes in HIV-1 infection.

BACKGROUND: Human Leukocyte Antigen (HLA) class I restricted Cytotoxic T Lymphocytes (CTLs) exert substantial evolutionary pressure on HIV-1, as evidenced by the reproducible selection of HLA-restricted immune escape mutations in the viral genome. An escape mutation from tyrosine to phenylalanine at the 135th amino acid (Y135F) of the HIV-1 nef gene is frequently observed in patients with HLA-A*24:02, an HLA Class I allele expressed in ~70% of Japanese persons. The selection of CTL escape mutations could theoretically result in the de novo creation of novel epitopes, however, the extent to which such dynamic "CTL epitope switching" occurs in HIV-1 remains incompletely known. RESULTS: Two overlapping epitopes in HIV-1 nef, Nef126-10 and Nef134-10, elicit the most frequent CTL responses restricted by HLA-A*24:02. Thirty-five of 46 (76%) HLA-A*24:02-positive patients harbored the Y135F mutation in their plasma HIV-1 RNA. Nef codon 135 plays a crucial role in both epitopes, as it represents the C-terminal anchor for Nef126-10 and the N-terminal anchor for Nef134-10. While the majority of patients with 135F exhibited CTL responses to Nef126-10, none harboring the "wild-type" (global HIV-1 subtype B consensus) Y135 did so, suggesting that Nef126-10 is not efficiently presented in persons harboring Y135. Consistent with this, peptide binding and limiting dilution experiments confirmed F, but not Y, as a suitable C-terminal anchor for HLA-A*24:02. Moreover, experiments utilizing antigen specific CTL clones to recognize endogenously-expressed peptides with or without Y135F indicated that this mutation disrupted the antigen expression of Nef134-10. Critically, the selection of Y135F also launched the expression of Nef126-10, indicating that the latter epitope is created as a result of escape within the former. CONCLUSIONS: Our data represent the first example of the de novo creation of a novel overlapping CTL epitope as a direct result of HLA-driven immune escape in a neighboring epitope. The robust targeting of Nef126-10 following transmission (or in vivo selection) of HIV-1 containing Y135F may explain in part the previously reported stable plasma viral loads over time in the Japanese population, despite the high prevalence of both HLA-A*24:02 and Nef-Y135F in circulating HIV-1 sequences.

Body composition change indices combined with Prognostic Nutritional Index predicts the clinical outcomes of patients with gastric cancer treated with immune checkpoint inhibitor.

OBJECTIVE: This study aimed to investigate the prognostic significance of the Prognostic Nutritional Index (PNI) in conjunction with body composition change indices, namely subcutaneous fat area (SFA) and skeletal muscle index (SMI), with regard to clinical outcomes in patients with gastric cancer (GC) undergoing immune checkpoint inhibitors (ICIs) treatment. METHODS: This retrospective investigation encompassed patients with comprehensive clinical and pathological data, inclusive of portal phase enhanced CT images. Continuous variables underwent analysis utilizing the Student t-test or Mann-Whitney U-test, while categorical variables were assessed employing the Pearson chi-squared test or Fisher test. Survival outcomes were evaluated using Kaplan-Meier survival curves and the Log-rank test. Independent prognostic indicators were determined through Cox regression analysis, and a nomogram predicting survival probability for progression-free survival (PFS) and overall survival (OS) was constructed. RESULTS: Within the PNI-SFA groups, patients in Group 1 exhibited inferior PFS and OS compared to the other two groups. Similarly, among the PNI-SMI groups, Group 1 patients demonstrated poorer PFS and OS. PNI-SMI and Eosi were identified as independent prognostic factors through Cox regression analysis. Furthermore, positive associations with patient prognosis were observed for BMI, SAF, SMI, and PNI. CONCLUSION: The comprehensive consideration of PNI-SFA and PNI-SMI proved to be a superior prognostic predictor for GC patients undergoing ICI treatment.

Influence of polymorphism in dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin-related (DC-SIGNR) gene on HIV-1 trans-infection.

The dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN) and DC-SIGN-related (DC-SIGNR) molecules on the cell surface are known to enhance human immunodeficiency virus type 1 (HIV-1) infection by capturing the virions and transmitting them to CD4+ T-cell, a process termed trans-infection. The neck region and carbohydrate recognition domain of the two proteins are important for efficient binding to the HIV-1 envelope protein. DC-SIGNR is polymorphic in Exons 4 and 5 that encode the neck region and carbohydrate recognition domain, respectively; the former contains a variable number of tandem repeats, and the latter the SNP (rs2277998). Since it remains unclear whether the DC-SIGNR polymorphism is related to the risk of HIV-1 infection, we tested possible effects of the polymorphism on HIV-1 trans-infection efficiency, by constructing six kinds of cDNAs encoding DC-SIGNR variants with various numbers of repeat units and various SNP. We were able to express the variants on the surface of Raji cells, a human B cell line. Flow cytometry showed that all the tested DC-SIGNR molecules were efficiently expressed on the cell surface at various levels; the assay for HIV trans-infection efficacy showed that all the tested variants had that activity with different efficacy levels. We found a correlation between the HIV trans-infection efficiency and the mean fluorescent intensity of DC-SIGNR expression (R(2)=0.95). Thus, our results suggest that the variation of the tested DC-SIGNR genotypes affects the efficacy of trans-infection by affecting the amounts of the protein expressed on the cell surface.

The prognostic value of systemic immune-inflammation index (SII) in patients after radical operation for carcinoma of stomach in gastric cancer.

BACKGROUND: The inflammatory reaction, which plays a key role in tumor microenvironment, is associated with the tumor development, progression and metastasis. We aim to estimate the potential prognostic value of systemic immune-inflammation index (SII) in patients after radical operation for carcinoma of stomach in gastric cancer. METHODS: One hundred and eighty-two patients with gastric cancer who underwent radical operation for carcinoma of stomach from January 2009 to December 2012 were enrolled and divided into a low SII group (<600×109 cells/L) and a high SII group (SII ≥600×109 cells/L) based on the optimal cutoff value of SII. The clinicopathological features were analyzed and compared between the two groups of patients. We analyzed the disease-free survival (DFS) and the overall survival (OS) by Kaplan Meier survival curve and log-rank test. The prognostic factors were used to evaluate by univariate analysis, and the independent prognostic factors were assessed by using multivariate Cox proportional hazards regression model. RESULTS: The median DFS and OS of all patients were 18.67 and 22.83 months, respectively. According to the univariate and multivariate analysis, the optimal cutoff value of SII had prognostic significance on DFS and OS. The median DFS and OS in low SII group were longer than those in high SII group (21.43 vs. 17.57 months, 28.03 vs. 18.30 months, respectively). The radical resection, type of surgery, metastatic lymph nodes ratio (MLNR), lymphocyte, SII, carcinoembryonic antigen (CEA) and Borrmann classification were the independent prognostic factors in gastric carcinoma. At the same time, the patients with MLNR in low SII group had longer DFS and OS than those with MLNR in high SII group, especially in patients with high MLNR. CONCLUSIONS: SII may serve as a convenient, low-cost and noninvasive prognostic marker for patients after radical operation for carcinoma of stomach in gastric cancer.

VWF, CXCL8 and IL6 might be potential druggable genes for acute coronary syndrome (ACS).

OBJECTIVE: Acute coronary syndrome (ACS) is currently a leading cause of morbidity and mortality worldwide. This study aimed to screen critical genes and miRNAs involved in ACS. MATERIALS AND METHODS: Microarray data (access number GSE19339) was downloaded from Gene Expression Omnibus (GEO) database. After data preprocessing, we screened the differentially expressed genes (DEGs) using limma package and subsequently performed enrichment analysis using DAVID tool. The protein-protein interaction (PPI) network and transcription factor (TF)-miRNA-target gene regulatory network were visualized using Cytoscape software. Finally, the drug-gene interactions were predicted using DGIdb database. RESULTS: A total of 425 DEGs were identified in ACS samples compared with healthy control samples. Functional enrichment analysis showed that DEGs were mainly involved in angiogenesis, inflammatory response and PI3K-Akt signaling pathway. IL6 and VEGFA were key nodes in PPI network. In addition, hsa-miR-29, hsa-miR-1, NFIC, NFKB1 and RELA were identified as key factors in TF-miRNA-target gene network. Finally, the prediction results revealed that VWF, CXCL8 and IL6 had higher degree than other genes. CONCLUSION: IL6 and VEGFA might play major roles in ACS progression. Two miRNAs (hsa-miR-29 and hsa-miR-1) and three TFs (NFIC, NFKB1 and RELA) were critical genes involved in pathological process of ACS. VWF, CXCL8 and IL6 might be potential druggable genes for ACS therapy.

[Marine Aerosol Using On-board Aerosol Mass Spectrometry].

Marine aerosols were measured in real-time by an on-board signal particle aerosol mass spectrometer(SPAMS) over the Southeast China Sea. The chemical compositions and size distribution characteristics of aerosol particles were obtained, and the sources and ion spectra were analyzed. The results showed that particle number concentration decreased with the distance apart from the coastal area. In the coastal area, the aerosol compositions were mainly determined by the emissions of industry, such as vessel, traffic and coal combustion etc. When it was far from the continent, aerosols were mainly affected by the sea sources. Aerosol particles during the observation period disturbed singly with a peak diameter value of 0.5 µm. Most of the particles were in the size range of 0.2 µm to 0.8 µm. High signal intensity of EC with high K+ intensity in the positive spectrum and HSO4- intensity in negative spectrum was present in the marine aerosol over the coastal area. However, the signals of NO3- and NO2- were absent in the negative spectrum. The signal intensity of EC was weak in the marine aerosol over the sea area far from the coastal area. High signal intensity of Na+ and weak Mg+,Ca+ and NaCl+ signals were present in the positive spectrum, while high signal intensity of MSA-,CN-,O- and HSO4- were present in negative spectrum which was considered to be the special ions spectrum of marine biological sources. It indicated that ambient aerosols over the observation area were influenced not only by the anthropogenic emission sources but also affected by the marine aerosol formation.

Rapid HIV-1 Disease Progression in Individuals Infected with a Virus Adapted to Its Host Population.

HIV-1 escape from CTL is predictable based on the Human Leukocyte Antigen (HLA) class I alleles expressed by the host. As such, HIV-1 sequences circulating in a population of hosts will harbor escape mutations specific to the HLA alleles of that population. In theory, this should increase the frequency of escape mutation transmission to persons expressing the restricting HLA allele, thereby compromising host immunity to the incoming HIV-1 strain. However, the clinical impact of infection with HIV-1 containing immune escape mutations has not conclusively been demonstrated. Japan's population features limited HLA diversity which is driving population-level HIV adaptation: for example, >60% of Japanese express HLA-A*24:02 and its associated Nef-Y135F escape mutation represents the population consensus. As such, Japan is an ideal population in which to examine this phenomenon. Here, we combine genetic and immunological analyses to identify A*24:02-positive individuals likely to have been infected with Y135F-containing HIV-1. Over a ~5 year follow-up, these individuals exhibited significantly lower CD4 counts compared to individuals inferred to have been infected with wild-type HIV-1. Our results support a significant negative clinical impact of pathogen adaptation to host pressures at the population level.

Influence of single-nucleotide polymorphisms in the multidrug resistance-1 gene on the cellular export of nelfinavir and its clinical implication for highly active antiretroviral therapy.

Protease inhibitors (PIs) such as nelfinavir (NFV) suppress HIV replication. PIs are substrates of P-glycoprotein (P-gp), the product of the multidrug-resistance-1 (MDR1) gene. Three single-nucleotide polymorphisms (SNPs) are present in exons of the MDR1 gene: MDR1 1236, MDR1 2677 and MDR1 3435. We speculated that these genetic polymorphisms affected PI concentration in the cell. To verify this hypothesis, we first genotyped these SNPs in 79 Japanese patients by the SNaPshot method and found incomplete linkage disequilibrium between the SNPs. Because the SNP at MDR1 3435 has been reported to be associated with P-gp expression, we evaluated the effect of that SNP on the export of NFV from HIV-positive patients' lymphoblastoid cell lines by measuring time-dependent decrease in the amount of intracellular NFV by high-performance liquid chromatography. We found the intracellular concentration of NFV in lymphoblastoid cell lines (LCLs) with the homozygous T/T genotype at MDR1 3435 were higher than that with C/C genotype with statistical significance. This suggests that the activity of P-gp in patients' LCL cells with the MDR1 3435 T/T genotype was lower. In a retrospective study we evaluated the effect of the SNPs on CD4 cell count recovery in response to antiretroviral treatment with PIs, and obtained statistically significant evidence that suggested marginal association of the SNP at MDR1 1236 but not at MDR1 2677 or MDR1 3435. As in vitro results were not consistent with the clinical evaluation, clinical importance of MDR1 genotyping for antiretroviral therapy remains to be investigated in a larger, case-controlled study.

Structure of TCR and antigen complexes at an immunodominant CTL epitope in HIV-1 infection.

We investigated the crystal structure of an HLA-A*2402-restricted CTL epitope in the HIV-1 nef gene (Nef134-10) before (pHLA) or after TCR docking. The wild type epitope and two escape mutants were included in the study. Y135F was an early-appearing major mutation, while F139L was a late-appearing mutation which was selected in the patients without Y135F. F139 was an eminent feature of the Nef134-10 epitope. Wild type-specific TCR was less fit to F139L mutant suggesting that F139L is an escape from the CTL against the wild type epitope. Although Y135F mutation disrupted the hydrogen bond to HLA-A*2402 His70, newly formed hydrogen bond between T138 and His70 kept the conformation of the epitope in the reconstituted pMHC. TCR from Y135F- or dually-specific CTL had unique mode of binding to the mutant epitope. Y135F has been reported as a processing mutant but CTL carrying structurally adequate TCR can be found in the patients.

Generation of rejuvenated antigen-specific T cells by reprogramming to pluripotency and redifferentiation.

Adoptive immunotherapy with functional T cells is potentially an effective therapeutic strategy for combating many types of cancer and viral infection. However, exhaustion of antigen-specific T cells represents a major challenge to this type of approach. In an effort to overcome this problem, we reprogrammed clonally expanded antigen-specific CD8(+) T cells from an HIV-1-infected patient to pluripotency. The T cell-derived induced pluripotent stem cells were then redifferentiated into CD8(+) T cells that had a high proliferative capacity and elongated telomeres. These "rejuvenated" cells possessed antigen-specific killing activity and exhibited T cell receptor gene-rearrangement patterns identical to those of the original T cell clone from the patient. We also found that this method can be effective for generating specific T cells for other pathology-associated antigens. Thus, this type of approach may have broad applications in the field of adoptive immunotherapy.