The prognostic value of tumor mutation burden (TMB) and its relationship with immune infiltration in breast cancer patients.

OBJECTIVE: Although the tumor mutation burden (TMB) was reported as a biomarker for immunotherapy of various cancers, whether it can effectively predict the survival prognosis in breast cancer patients remains unclear. In this study, the prognostic value of TMB and its correlation with immune infiltration were explored by using multigroup studies. METHODS: The somatic mutation data of 986 breast cancer patients were obtained from TCGA database. Breast cancer patients were divided into a low-TMB group and a high-TMB group according to the quartile of TMB scores. The differentially expressed genes (DEGs) were identified by the "limma" R program. The CIBERSORT algorithm was utilized to estimate the immune cell fraction of each sample. The TIMER database was utilized to evaluate the association between CNVs of immune genes and tumor immune cell infiltration and the prognostic value of the immune cells in breast cancer. RESULTS: In breast cancer, TP53, PIK3CA, TTN, CDH1 and other genes were the most important mutated genes. Higher survival rate of patients was found in the low-TMB group. Among the top 10 DEGs, three of them belong to the KRT gene family. GSEA enrichment analysis showed that MAPK, Hedgehog, mTOR, TGF-bate and GnRH signaling pathways were enriched in the low-TMB group. The infiltration levels of the most of immune cells were higher in the low-TMB group (P < 0.01). Higher expression of CCL18 and TRGC1 was correlated with poor prognosis. Breast cancer patients with CCL18 copy number variations, especially arm-level gains, showed significantly decreased immune cell infiltration. In the low B cell infiltration group, the survival prognosis of breast cancer patients was poor. CONCLUSIONS: TMB is a potential prognosis marker in breast cancer. Immune-related gene CCL18 and TRGC1 are biomarkers of poor prognosis while immune (B cell) infiltration is a biomarker of good prognosis.

Identification of genes and pathways leading to poor prognosis of non-small cell lung cancer using integrated bioinformatics analysis.

Background: Non-small cell lung cancer (NSCLC) is a common malignancy with a high morbidity and mortality rate worldwide, but the driver genes and signaling pathways involved are largely unclear. Herein, our study aimed to identify significant genes with poor outcome and underlying mechanisms in NSCLC using bioinformatics analyses. Methods: Gene expression profiles (GSE33532, GSE19188, GSE102287, GSE27262), including 319 NSCLC and 232 adjacent lung tissues, were downloaded from the GEO database. Differentially expressed genes (DEGs) were identified by the GEO2R online tool. Functional and pathway enrichment analyses were performed via the DAVID database. The protein-protein interactions (PPIs) of these DEGs were constructed by the STRING website and visualized by the Cytoscape software platform. The expression of hub genes in NSCLC was validated through the GEPIA database. Kaplan-Meier plotter was used to analyse the survival rate with multivariate Cox regression. The expression of protein tyrosine kinase 2 (PTK2) in NSCLC and adjacent lung tissues was evaluated on the UALCAN database platform. Results: A total of 225 significant DEGs were obtained between NSCLC and adjacent lung tissues, containing 52 upregulated genes and 173 downregulated genes. The DEGs were clustered based on functions and signaling pathways that may be closely associated with NSCLC occurrence. A total of 174 DEGs were identified from the PPI network complex. Top 10 hub genes were selected by CytoHubba plugin. As independent predictors, seven genes (COL1A1, ADAM12, VWF, OGN, EDN1, CAV1, ITGA8) were associated with poor prognosis in NSCLC via multivariate Cox regression (P<0.01). Four genes (VWF, CAV1, ITGA8, COL1A1) were found to be significantly enriched in the focal adhesion pathway (P=1.04E-04) and to be upstream regulators of PTK2. PTK2 was upregulated in NSCLC and associated with poor survival prognosis in lung squamous cell carcinoma (LUSC). Conclusions: Taken together, the important genes and pathways in NSCLC were identified by using integrated bioinformatics analysis. PTK2 could be a key gene associated with the biological process of NSCLC formation and progression and a potential therapeutic target for NSCLC treatment.

The Predictive Value of Red Cell Distribution Width and Red Cell Distribution Width to Erythrocyte Count Ratio for Adverse Cardiovascular Events During the Hospitalization of Patients of ST-segment Elevation Myocardial Infarction.

BACKGROUND: High red cell distribution width (RDW) is correlated with poor prognosis in acute coronary syndromes (ACS), including ST-segment elevation myocardial infarction (STEMI). However, the association of red cell distribution width to erythrocyte count ratio (RER) with STEMI undergoing percutaneous coronary intervention (PCI) during hospitalization has not been investigated. Therefore, we performed a retrospective study to investigate whether RER is associated with STEMI patients after PCI during hospitalization. METHODS: A total of 331 patients, who were hospitalized for STEMI and underwent PCI, were enrolled. Receiver operating characteristic curve (ROC) analyses were used to find the cutoff value of RER and classify the patients into two groups including higher RER group and lower RER group by cutoff value. Differences between measured parameters in higher RER and lower RER groups were analyzed by the Mann-Whitney U test. The evaluation correlation of RDW, red blood cell, and RER with major adverse cardiovascular events was determined by bivariate regression analysis. Univariate logistic regression analysis was used to determine the factors associated with adverse cardiovascular events during the hospitalization of STEMI patients. Multivariate logistic regression analysis was performed to evaluate the potential independent predictors of STEMI. RESULTS: According to ROC analysis, the cutoff value of RER and RDW is 3.10 and 13.9, the sensitivity is 51% and 35%, the specificity is 76% and 80%, respectively. RER showed improved diagnostic capacity compared to RDW in correlation with adverse cardiovascular events during hospitalization in STEMI patients (p < 0.001). Compared with the lower RER group, the incidence of adverse cardiovascular events in STEMI patients is elevated in the higher RER group (75% vs. 64.5%, p < 0.05). Bivariate regression analysis indicated that RER and RDW showed a good correlation with adverse cardiovascular events, and the difference was statistically significant (R = 0.10 p < 0.05 vs. R = 0.05 p < 0.05). Univariate logistic regression analysis showed that age, heart rate, left ventricular ejection fraction, hyperlipidemia, RDW, mean platelet volume, total cholesterol, and RER were correlated with the occurrence of adverse cardiovascular events during the hospitalization of STEMI patients (p < 0.05). Multivariate logistic regression analysis demonstrated that RER could be an independent predictor of adverse cardiovascular events in STEMI patients (B: 0.574, OR: 1.776, 95% CI: 1.043 ~ 3.023, p < 0.05). CONCLUSIONS: RER and RDW demonstrated good correlation with adverse cardiovascular events during hospitalization in STEMI patients. RER is a potential independent predictor of adverse cardiovascular events during hospitalization in STEMI patients.

Prediction of MiR-21-5p in Promoting the Development of Lung Adenocarcinoma via PDZD2 Regulation.

BACKGROUND Lung adenocarcinoma currently accounts for the highest cancer-related mortality rate worldwide. MiR-21-5p has a vital role in various types of cancers. We have analyzed the miR-21-5p expression level, prognosis, and associated molecular pathways in lung adenocarcinoma with multiple bioinformatics databases. MATERIAL AND METHODS The Cancer Genome Atlas (TCGA) database was employed to fetch the miR-21-5p expression profile in multiple tumors. We used the UALCAN platform to assess the differential regulation of the miR-21-5p in healthy tissue and lung adenocarcinoma. Also, the survival prognosis of the miR-21-5p in each stage of lung adenocarcinoma was done by the Kaplan-Meier database. The STARBASE and UALCAN databases were employed to predict the miR-21-5p target genes, and the levels of target genes and their prognostic value were analyzed. RESULTS MiR-21-5p was overexpressed in the majority of human cancers. MiR-21-5p demonstrated escalated expression in the lung adenocarcinoma tissue in contrast to the normal tissue (P<0.05). Poor prognosis was witnessed in the miR-21-5p high expression group as compared to the low expression group (hazard ratio [HR]= 1.59, P<0.05). PDZD2 was predicted as a miR-21-5p potential target. We found a negative correlation between PDZD2 and miR-21-5p (r=-0.255, P<0.05). PDZD2 was downregulated in lung adenocarcinoma (P<0.05). Overexpression of PDZD2 was associated with a better prognosis of survival in lung adenocarcinoma patients (HR=0.45, P<0.05). CONCLUSIONS MiR-21-5p exhibits the potential to act as a biomarker for the survival prognosis of lung adenocarcinoma. It might be responsible for the onset and progression of lung adenocarcinoma through PDZD2 regulation.

A Potential Serum Biomarker, Albumin-to-Glutamyltransferase Ratio, Suggests the Severity of Liver Disease.

BACKGROUND: We aimed to investigate the utility of albumin-to-glutamyltransferase ratio (AGR) as a new biomarker to distinguish hepatic carcinoma from hepatitis, as early disease diagnosis, prognosis or monitoring could improve patient management and outcomes. METHODS: Clinical characteristics of 34 hepatitis (women 19), 88 cirrhosis (women 22) and 52 hepatic carcinoma (women 9) cases were retrospectively reviewed. Patients diagnosed with cirrhosis were classified by Child-Pugh score and the presence of ascites. The differences among groups were evaluated by the Kruskal-Wallis test and Mann-Whitney U. The linear correlation between variables was assessed by Spearman's correlation analysis. The diagnostic value of albumin-to-glutamyltransferase (AGR) was considered using receiver operating characteristic (ROC) curves. Multiple logistic regression analysis and univariate logistic regression analysis were used to identify AGR as an independent predictor in liver disease progression. RESULTS: The significant differences among the hepatitis vs. cirrhosis vs. and hepatic carcinoma were AST (108.50 ± 184.00 vs. 38.00 ± 21.50 vs. 47.00 ± 71.00, p < 0.01), TP/AST (TAR, 0.67 ± 0.69 vs. 1.77 ± 0.87 vs. 1.36 ± 0.95, p < 0.01), and ALB/GGT (AGR, 0.32 ± 0.27 vs. 0.67 ± 0.43 vs. 0.20 ± 0.26, p < 0.05). At the same time, AST (32.00 ± 13.50 vs. 53.00 ± 23.00 vs. 114.50 ± 42.50, p < 0.05) and TAR (2.15 ± 0.72 vs. 1.28 ± 0.74 vs. 0.64 ± 0.39, p < 0.05) were higher but AGR (0.86 ± 0.54 vs. 0.46 ± 0.32 vs. 0.26 ± 0.22, p < 0.05) was lower in Child-Pugh class C group compared with group B and C. TAR (1.92 ± 0.73 vs. 0.98 ± 0.89, p < 0.01) and AGR (0.79 ± 0.52 vs. 0.46 ± 0.28, p < 0.05) were significantly elevated in the serum of cirrhosis with no ascites compared with the cirrhosis patients suffered from ascites, while AST (35.00 ± 14.50 vs. 63.00 ± 44.50, p < 0.01) was reduced in cirrhosis patients with no ascites. Furthermore, AST (r = 0.4490, p<0.01) was positively correlated with AFP, TAR (r = -0.4393, p < 0.01) and AGR (r = -0.4395, p < 0.01) were negatively correlated with AFP. The ROC curve analysis for AST had an area under the curve (AUC) ranging from 0.66 to 0.82, TAR ranged from 0.64 to 0.80 and AGR ranged from 0.54 to 0.72. Multiple logistic regression analysis revealed AGR as an independent parameter to distinguish liver can¬cer to hepatitis, and AGR was associated with the presence of ascites and the progression in cirrhosis patients. CONCLUSIONS: AGR is a potential biomarker for diagnosis of liver disease progression.

The Relationship between Erythrocyte Sedimentation Rate and Erythrocyte Sedimentation/Red Blood Cell Ratio and Disease Activity in Systemic Lupus Erythematosus.

BACKGROUND: The treatment options of systemic lupus erythematosus (SLE) patients in active and inactive phases are very different clinically, and the prognosis of patients with active SLE is much worse than inactive patients. However, the present indicators for diagnosis of SLE in activity are limited and inefficient. METHODS: Three hundred thirty patients with SLE were included. All patients are classified as SLEDAI (systemic lupus erythematosus disease activity index) > 4 as active and SLEDAI ≤ 4 as inactive. The linear correlation between variables was assessed by Pearson's correlation analysis. The difference between parameters in active and inactive patients was evaluated by the Mann-Whitney U test. The evaluation capacity of erythrocyte sedimenta-tion/red blood cell (ERR) and red blood cell/albumin ratio (RAR) on SLE activity was determined by bivariate regression analysis. Sensitivity and specificity are assessed by receiver operating characteristic curve (ROC). RESULTS: Compared with the inactive SLE, ESR (52.97 ± 35.66 vs. 32.38 ± 29.16 p < 0.001), ERR (15.40 ± 12.41 vs. 8.19 ± 8.10 p < 0.001) and RAR (0.13 ± 0.10 vs. 0.11 ± 0.20 p = 0.038) are all elevated in active SLE (52.97 ± 35.66 vs. 32.38 ±2 9.16 p < 0.001). ERR shows better correlation than RAR with ESR (p < 0.001 vs. p = 0.911). Patients with active SLE exhibited higher SLEDAI than those with inactive SLE (8.67 ± 2.67 vs. 3.27 ± 1.36, p < 0.001). According to ROC analysis, when ESR levels > 58.5 and ERR levels > 13.18, the sensitivity is 37.6% and 45.2%, the specificity is 83.0% and 83.2%. CONCLUSIONS: ESR and ERR are potential indicators for diagnosis of active and inactive SLE.

[Plasma miRNA-23a and miRNA-451 as candidate biomarkers for early diagnosis of nonsmall cell lung cancer: a case-control study].

OBJECTIVE: To study the value of plasma miRNA23-a and miRNA-451 as potential biomarkers for early diagnosis of non-small cell lung cancer (NSCLC). METHODS: Fifty patients with NSCLC and 50 healthy control subjects were recruited for testing the plasma levels of miRNA23-a and miRNA-451 and their expression levels in the tumor tissues using qRT-PCR. The correlations of the plasma levels of miRNA23-a and miRNA-451 with their expressions in the tumor tissues were analyzed. The diagnostic power of CEA, miRNA23-a and miRNA-451 for NSCLC was evaluated using the receiver-operating characteristics (ROC) curves and the area under the ROC curves (AUC). In the NSCLC cell line A549, we tested the effect of inhibition of miRNA-23a and miRNA-451 on the expression levels of SPRY2 and MIF mRNA using qRT-PCR. RESULTS: The expression levels of miRNA-23a and miRNA-451 in NSCLC tissues was significantly associated with smoking, tumor size, lymph node metastasis and TNM stage (P < 0.05). Compared with those in the control group, miRNA-23a level was significantly increased while miRNA-451 was significantly down-regulated in the tumor tissues and plasma of NSCLC patients. The plasma levels of miRNA-23a and miRNA-45 were strongly correlated with their expression levels in the tumor tissues. ROC analysis showed that for the diagnosis of NSCLC, the AUC, sensitivity and specificity of either miRNA-23a or miRNA-451 were significantly higher than those of CEA (P < 0.05). The combination of miRNA23-a and miRNA-451 markedly improved the AUC (0.900), sensitivity (78%) and specificity (86%) for the diagnosis. In A549 cells, inhibition of miRNA23-a and miRNA-451 resulted in significantly increased expression levels of SPRY2 mRNA and MIF mRNA, respectively. CONCLUSIONS: miRNA-23a and miRNA-451 can be used as potential biomarkers for early diagnosis of NSCLC, and their combined detection can be more effective for the diagnosis.

MicroRNA-194 inhibition improves dietary-induced non-alcoholic fatty liver disease in mice through targeting on FXR.

Non-alcoholic fatty liver disease (NAFLD) affects obesity-associated metabolic syndrome, which exhibits hepatic steatosis, insulin insensitivity and glucose intolerance. Previous studies indicated that hepatic microRNAs (miRs) play critical roles in the development of NAFLD. In this study, we aim to explore the pathophysiological role of miR-194 in obesity-mediated metabolic dysfunction. Our findings show that the high fat diet or palmitic acid treatment significantly increase hepatic miR-194 levels in vivo and in vitro. Silence of miR-194 protects palmitic acid-induced inflammatory response in cultured hepatocytes, and attenuates structural disorders, lipid deposits and inflammatory response in fatty liver. MiR-194 inhibitor also improves glucose and insulin intolerance in obese mice. Through dual luciferase assay, we demonstrate that miR-194 directly binds to FXR/Nr1h4 3'-UTR, and inhibits gene expression of FXR/Nr1h4. Furthermore, overexpression of miR-194 downregulates FXR/Nr1h4 in cultured hepatocytes, but miR-194 inhibitor reversely increases FXR/Nr1h4 expression in obese mouse liver tissues. On the contrast, silence of FXR/Nr1h4 abolishes the hepatic benefits in obese mice treated with miR-194 inhibitor. Present study provides a novel finding that suppression of miR-194 attenuates dietary-induced NAFLD via upregulation of FXR/Nr1h4. The findings suggest miR-194/FXR are potential diagnostic markers and therapeutic targets for NAFLD.

Discriminating Value of Calprotectin in Disease Activity and Progression of Nonradiographic Axial Spondyloarthritis and Ankylosing Spondylitis.

It has been controversial whether ankylosing spondylitis (AS) and nonradiographic axial spondyloarthritis (nr-axSpA) are separate or different phases of radiographic progression. We determined that serum calprotectin level (ng/ml) was higher in AS (15.30 ± 6.49) and nr-axSpA (17.76 ± 8.59) patients than in healthy individuals (7.40 ± 2.67). No difference was observed in calprotectin level between these two groups. Elevated calprotectin was positively correlated with ESR, CRP, BASDAI, and ASDAS as well as SPARCC scoring and had no correlation with BASFI and mSASSS. No correlation was observed between calprotectin and Wnt/ß-catenin pathway markers. Serum calprotectin can be used as a marker for inflammation in both nr-axSpA and AS, while it does not contribute to the discrimination of AS and nr-axSpA. Calprotectin-mediated inflammation was not correlated with principle effectors of Wnt/ß-catenin pathway, indicating that inflammation and bone fusion might be separate processes of the disease.

[Association of colorectal adenoma and metabolic syndrome and relevant parameters].

OBJECTIVE: To evaluate the association of colorectal adenoma with metabolic syndrome (MS) and relevant parameters. METHODS: Clinical data of 289 subjects who underwent screening colonoscopy in the University of Hong Kong-Shenzhen Hospital from January 2014 to June 2015 were retrospectively analyzed, including 130 normal subjects (normal group) and 159 cases with colorectal adenoma confirmed by pathology(adenoma group). Levels of MS-associated parameters were compared between the two groups, and the association of metabolic diseases with colorectal adenoma was examined. RESULTS: The gender, smoking and drinking habit, regular physical activity, family history of colorectal cancer, and consumption history of long-term non-steroidal anti-inflammatory drugs were not significantly different between two groups (all P>0.05). As compared to normal group, adenoma group had higher body mass index (BMI) [(23.5±3.2) kg/m(2) vs. (22.7±2.8) kg/m(2), t=1.97, P=0.050], larger abdominal circumference [(83.4±10.3) cm vs. (79.6±13.8) cm, t=2.46, P=0.015], higher serum high-density lipoprotein level [(1.3±0.3) mmol/L vs. (1.2±0.3) mmol/L, t=2.03, P=0.044], and higher serum cholesterol [(5.4±1.0) mmol/L vs. (5.0±1.1) mmol/L, t=2.39, P=0.018]. No significant difference was demonstrated in comparing hip circumference and waist-hip ratio, as well as serum fasting glucose and triglyceride(all P>0.05). Higher incidence of colorectal adenoma was found in subjects with MS [69.8%(37/53) vs. 1.7%(122/236), P=0.017], overweight or obesity [65.1% (56/86) vs. 50.7%(103/203), P=0.025], hypertension [67.3%(37/55) vs. 52.1%(122/234), P=0.046] and hypercholesterolemia [66.7%(64/96) vs. 49.2%(95/193), P=0.005]. CONCLUSIONS: Metabolic syndrome increased the risk of developing colorectal adenoma. The mechanism may be related to higher serum cholesterol and high density lipoprotein, which may lead to the elevated catabolism of serum cholesterol. Screening colonoscopy should be performed for patients diagnosed as metabolic syndrome, especially for those with central obesity and hypercholesterolemia, thus early diagnosis and treatment of colorectal adenoma may be available.