Study on the Prognostic Values of TTC36 Correlated with Immune Infiltrates and Its Methylation in Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) remains an incurable disease with a very poor clinical outcome. The purpose of this article was to investigate whether the expression or methylation of tetrapeptide repeat domain 36 (TTC36) could be used as a prognostic marker in hepatocellular carcinoma. TCGA database was used to obtain information on HCC gene expression and the associated clinical features of HCC patients. Differentially expressed genes (DEGs) were screened between 374 HCC specimens and 50 nontumor specimens. The expression and prognostic value of TTC36 were analyzed. The correlations between TTC36 and cancer immune infiltrates were investigated via TIMER. In this study, HCC specimens and nontumor specimens were compared and 35 DEGs were found between them. Among the 35 DEGs, the expression of TTC36 was significantly reduced in HCC samples compared with nontumor samples. Survival tests revealed that patients with low TTC36 expression had a shorter overall survival than patients with high TTC36 expression. TTC36 was found to be an independent predictive factor for HCC in both univariate and multivariate regression analyses. TTC36 was negatively regulated by TTC36 methylation, leading to its low expression in HCC tissues. Immune analysis revealed that TTC36 expression has significant correlations with B cell, T cell CD4+, neutrophil, macrophage, and myeloid dendritic cell. Finally, TTC36 expression was dramatically reduced in HCC cells, and overexpression greatly suppressed HCC cell proliferation and invasion, according to our experimental results. Overall, our data suggested that TTC36 could be applied as a prognostic marker for predicting outcome and immune infiltration in HCC.

Differential microRNA expression profile of Trichinella spiralis larvae after exposure to the host small intestinal milieu.

The process by which Trichinella spiralis muscle larvae are activated to infect the intestine after exposure to the host small intestinal milieu is crucial for the successful establishment of T. spiralis infection. However, the molecular mechanism underlying the invasion of intestinal epithelial cells by T. spiralis has not been elucidated. MicroRNAs are a class of small noncoding RNAs that participate in parasite growth and development, pathogenic processes, and host-parasite interactions. In the present study, the differential expression profile of miRNAs in T. spiralis after exposure to the mouse small intestinal milieu was analysed using Solexa high-throughput sequencing technology. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed to analyse the functions of miRNA target genes, and dual-luciferase reporter assays were subsequently applied to test the regulatory effects of one significantly decreased miRNA (let-7) on its four predicted target genes. In total, 2,000 known miRNAs (930 upregulated and 1070 downregulated) and 43 novel miRNAs (22 upregulated and 21 downregulated) were found to be differentially expressed in intestinal larvae, compared with muscle larvae. The KEGG pathway analysis showed that the predicted target genes of differentially expressed miRNAs were involved in 299 different pathways, and the top 10 pathways were metabolic pathways, biosynthesis of secondary metabolites, neuroactive ligand-receptor interaction, lysosome, focal adhesion, purine metabolism, starch and sucrose metabolism, tight junction, carbohydrate digestion and absorption, and pathways in cancer. As one of the most widely studied miRNA families, the expression of let-7 was significantly decreased in T. spiralis after exposure to host small intestinal milieu. A dual-luciferase reporter assay revealed that neuropeptide Y receptor type 6 and carboxypeptidase E were direct target genes of let-7, and were downregulated by binding with their 3' UTR. GO function analysis showed that carboxypeptidase E had multiple enzymatic activities, suggesting that it might participate in cell membrane damage and larval invasion. These data suggest that the differentially expressed miRNAs in T. spiralis might have a regulatory role in the invasion of host intestinal epithelial cells. This study provides a new insight into the molecular mechanisms of invasion by T. spiralis and the regulatory functions of miRNAs in host-Trichinella interactions.

Identification of key genes associated with progression and prognosis for lung squamous cell carcinoma.

BACKGROUND: Lung squamous cell carcinoma (LUSC) is a major subtype of lung cancer with limited therapeutic options and poor clinical prognosis. METHODS: Three datasets (GSE19188, GSE33532 and GSE33479) were obtained from the gene expression omnibus (GEO) database. Differentially expressed genes (DEGs) between LUSC and normal tissues were identified by GEO2R, and functional analysis was employed using the Database for Annotation, Visualization and Integrated Discovery (DAVID) online tool. Protein-protein interaction (PPI) and hub genes were identified via the Search Tool for the Retrieval of Interacting Genes (STRING) and Cytoscape software. Hub genes were further validated in The Cancer Genome Atlas (TCGA) database. Subsequently, survival analysis was performed using the Kapla-Meier curve and Cox progression analysis. Based on univariate and multivariate Cox progression analysis, a gene signature was established to predict overall survival. Receiver operating characteristic curve was used to evaluate the prognostic value of the model. RESULTS: A total of 116 up-regulated genes and 84 down-regulated genes were identified. These DEGs were mainly enriched in the two pathways: cell cycle and p53 signaling way. According to the degree of protein nodes in the PPI network, 10 hub genes were identified. The mRNA expression levels of the 10 hub genes in LUSC were also significantly up-regulated in the TCGA database. Furthermore, a novel seven-gene signature (FLRT3, PPP2R2C, MMP3, MMP12, CAPN8, FILIP1 and SPP1) from the DEGs was constructed and acted as a significant and independent prognostic signature for LUSC. CONCLUSIONS: The 10 hub genes might be tightly correlated with LUSC progression. The seven-gene signature might be an independent biomarker with a significant predictive value in LUSC overall survival.

[Analysis of (DPY19L2 gene variant in two brothers affected with globozoospermia].

OBJECTIVE: To explore the molecular basis for two brothers affected with globozoospermia. METHODS: Whole exome sequencing was carried out for both patients. Candidate variant was verified by Sanger sequencing and quantitative real-time PCR (qRT-PCR). RESULTS: Whole exome sequencing, Sanger sequencing and qRT-PCR verification revealed a heterozygous c.384dup (p.Glu129*) variant in the DPY19L2 gene in the two brothers and their mother. A large heterozygous deletion, spanning approximately 164.5 kb and encompassing the entire DPY19L2 gene, was detected on chromosome 12 of the two patients and their father. CONCLUSION: The c.384dup (p.Glu129*) variant and deletion of the DPY19L2 gene probably underlie the pathogenesis of globozoospermia in the two patients, which was in keeping with the autosomal recessive inheritance of disease in this pedigree.

Differential Expression and Diagnostic Significance of Pre-Albumin, Fibrinogen Combined with D-Dimer in AFP-Negative Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is one of the most malignant cancers with high morbidity and mortality. Nowadays, AFP-negative hepatocellular carcinoma (AFP-NHCC) has been found in many HCC patients and AFP analysis can't be used to screen HCC in these cases. In this study, we have examined the expression patterns of pre-albumin (PA), fibrinogen, D-Dimer and their clinical significance in AFP-NHCC. We recruited 214 AFP-NHCC patients and 210 controls in the study. PA, fibrinogen and D-Dimer levels were detected by turbidimetry, clauss and immunoturbidimetry methods, respectively. Serum PA levels were significantly lower in AFP-NHCC (84.5 ± 24.7 mg/L) than that in the controls (240.6 ± 59.4 mg/L, P < 0.05). For plasma fibrinogen levels, there was no difference between the controls (2.9 ± 0.7 g/L) and AFP-NHCC (2.5 ± 0.7 g/L). Compared with AFP-NHCC (0.8 ± 0.2 mg/L), plasma D-Dimer levels were significantly lower in controls (0.1 ± 0.0 mg/L, P < 0.05). The levels of PA, fibrinogen and D-Dimer were significantly correlated with differentiation (P < 0.01), and the PA and D-Dimer values were correlated with TNM stage (P < 0.05). Moreover, PA levels were correlated with tumor size (P = 0.034). Receiver operating characteristic curve (ROC) analyses elaborated that combination of PA, fibrinogen and D-Dimer possessed a higher sensitivity (93.4%) for differentiating AFP-NHCC from the controls, but the diagnostic specificity was reduced due to the combination of fibrinogen. After adjusting for all significant outcome predictors of the univariate logistic regression analysis, low levels of PA and high levels of D-Dimer were remained independent unfavorable outcome predictors (P < 0.05). Our data suggested that the expression levels of PA, fibrinogen and D-Dimer played critical roles in AFP-NHCC tumorigenesis. Moreover, PA and D-Dimer might be considered as potential diagnostic indicators in AFP-NHCC.

Preoperative Imaging and Microscopic Navigation During Surgery Can Avoid Unnecessarily Opening the Mastoid Air Cells Through Craniotomy Using the Retrosigmoid Approach.

OBJECTIVE: To analyze treatment of microvascular decompression using the retrosigmoid approach (RA) in primary trigeminal neuralgia and hemifacial spasm using preoperative images combined with intraoperative microscopic navigation to avoid unnecessarily opening the mastoid air cells (MACs). METHODS: Ten patients with primary trigeminal neuralgia and 20 patients with hemifacial spasm (test group) were treated using RA for microvascular decompression. Preoperative head magnetic resonance angiography and temporal bone computed tomography were performed and the images registered using SPM12 and fused with MRIcron to determine the relationship between MACs and sigmoid sinuses. An O-arm was used for navigation, and the transverse sigmoid sinus was projected under a microscope to guide RA. A control group comprised 139 patients who had the same surgical procedure as the test group but without image processing or intraoperative navigation. RESULTS: The relationship between MACs and the ipsilateral sigmoid sinus was classified as follows: I, MACs did not exceed the lateral edge of the ipsilateral sigmoid sinus (10/60); II, MACs exceeded the ipsilateral lateral edge of the sigmoid sinus but did not exceed the medial edge (42/60); and III, MACs exceeded the medial edge of the ipsilateral sigmoid sinus (8/60). Test and control groups showed significant differences in the incidences of opening MACs (P = 0.003). There was no cerebrospinal fluid leakage or scalp and intracranial infection at follow-up. CONCLUSIONS: Image processing and intraoperative microscopic navigation can avoid unnecessarily opening MACs and might reduce postoperative cerebrospinal leakage and scalp infection after RA craniotomy.

miR-135a-5p and miR-124-3p Inhibit Malignancy of Glioblastoma by Downregulation of Syndecan Binding Protein.

Due to its high invasiveness, glioblastoma is difficult to treat by surgery, radiotherapy, chemotherapy or any combination therapy. Syndecan binding protein (SDCBP), a widely distributed intracellular scaffold protein, has an important role in both physiological and pathological process. In the current work, we have identified target sequences for miR-135a-5p and miR-124-3p in the 3'-untranslated region of the SDCBP mRNA. Therefore, we have investigated the relationship between SDCBP, miR-135a-5p and miR-124-3p in glioblastoma multiforme cells T98G and U87 in vitro and in vivo. Dual luciferase activity assay documented that SDCBP is, in fact, the target of miR-135a-5p, miR-124-3. Western blot, qRT-PCR, proliferation, migration, and invasion assays have demonstrated that of silencing SDCBP or overexpressing miR-135a-5p/miR-124-3p significantly interferes with the malignant properties of glioblastoma cells. In vivo studies have shown that silencing SDCBP or overexpressing miR-135a-5p/miR-124-3p result in a marked decrease of tumor size and prolong life of tumor-bearing mice. A therapy combining the three treatments inhibits synergistically subcutaneous tumor growth in nude mice. In conclusion, proliferation, migration and invasion of glioblastoma can be inhibited by targeted regulation of SDCBP through upregulation of miR-135a-5p and miR-124-3p.

The diagnostic and prognostic significance of long noncoding RNAs expression in thyroid cancer: A systematic review and meta-analysis.

OBJECTIVE: Thyroid cancer (TC) is the most common malignant endocrine-related cancer with an increasing trend worldwide. Therefore, it's in urgent need to find new markers for prognosis and diagnosis. Many long noncoding RNAs (lncRNAs) have been reported to be aberrantly expressed in TC, and may serve as biomarkers. Therefore, we performed this meta-analysis to systematically summarize the relationship between lncRNA expressions and TC. METHODS: Sources from PubMed, Embase and Web of Science were searched. A total of 16 eligible studies including 15 on clinicopahology, 5 on prognosis and 6 on diagnosis were enrolled in our meta-analysis. Revman5.3 and Stata11.0 Software were used to conduct the meta-analysis. RESULTS: For diagnostic value, lncRNAs could discriminate between TC and the normal, and yield a high overall sensitivity and specificity (0.80, 95% CI: 0.75-0.84; 0.80, 95% CI: 0.70-0.87). Meanwhile, their sensitivity and specificity were 0.74 (95% CI: 0.59-0.85) and 0.81 (95% CI: 0.73-0.88) respectively, when used to differentiate patients with lymph node metastasis (LNM) from without LNM. The summary receiver operator characteristic curve (sROC) showed that lncRNAs could be considered as valuable diagnostic markers for distinguishing TC patients from the normal (AUC = 0.84) and TC patients with LNM from TC patients without LNM (AUC = 0.85). CONCLUSIONS: In summary, our meta-analysis suggested that lncRNAs could function as potential diagnostic markers for TC and predict the LNM. In addition, the systematic review elaborated that lncRNAs might be as prognostic indicators in TC.

The platelet-to-lymphocyte ratio, superior to the neutrophil-to-lymphocyte ratio, correlates with hepatitis C virus infection.

OBJECTIVES: The platelet-to-lymphocyte ratio (PLR) and neutrophil-to-lymphocyte ratio (NLR) have been studied widely in cancer diseases. However, their correlation with hepatitis C virus (HCV) infection is unknown. The aim of this study was to investigate the correlation of PLR and NLR with disease severity in patients with HCV-related liver disease and the virological response in chronic hepatitis C (CHC) patients. METHODS: The clinical data of 120 HCV-infected patients and 40 healthy controls were analyzed. The clinical data of 24 CHC patients who had been followed up regularly were collected for the following time points: before treatment (week 0) and weeks 4, 48, and 72 during treatment. These data were also analyzed. All data were collected from the database of the hospital patient electronic medical record system. RESULTS: The HCV-related cirrhosis group and HCV-related hepatocellular carcinoma group were found to have lower PLRs (61±31 and 51±23) than the healthy controls (115±23). The PLR of the HCV cleared group (154±85) was significantly higher than that of the HCV untreated group and HCV uncleared group (90±28 and 88±40, respectively). Receiver operating characteristics curve analysis for the PLR showed an area under the curve of 0.772 (95% confidence interval 0.674-0.869, p<0.000); for NLR, the area under the curve was 0.612 (95% confidence interval 0.495-0.730, p=0.063). Furthermore, an increasing PLR in CHC patients indicated a good virological response, and a stable PLR or a downward trend in PLR could predict no rapid virological response being achieved by week 4, and even no sustained virological response by week 72. CONCLUSIONS: The PLR is closely related to disease severity in patients with HCV-related liver disease and to the virological response in CHC patients. Dynamic continuous monitoring of the PLR will contribute to disease surveillance, with an increasing tendency predicting a good virological response.

Targeting PDK1 with dichloroacetophenone to inhibit acute myeloid leukemia (AML) cell growth.

Pyruvate dehydrogenase kinase-1 (PDK1), a key metabolic enzyme involved in aerobic glycolysis, is highly expressed in many solid tumors. Small molecule compound DAP (2,2-dichloroacetophenone) is a potent inhibitor of PDK1. Whether targeting PDK1 with DAP can inhibit acute myeloid leukemia (AML) and how it works remains unknown. In this study, we evaluated the effect of inhibition of PDK1 with DAP on cell growth, apoptosis and survival in AML cells and identified the underlying mechanisms. We found that treatment with DAP significantly inhibited cell proliferation, increased apoptosis induction and suppressed autophagy in AML cells in vitro, and inhibited tumor growth in an AML mouse model in vivo. We also showed that inhibition of PDK1 with DAP increased the cleavage of pro-apoptotic proteins (PARP and Caspase 3) and decreased the expression of the anti-apoptotic proteins (BCL-xL and BCL-2) and autophagy regulators (ULK1, Beclin-1 and Atg). In addition, we found that DAP inhibited the PI3K/Akt signaling pathway. Furthermore, we demonstrated that PDK1 interacted with ULK1, BCL-xL and E3 ligase CBL-b in AML cells, and DPA treatment could inhibit the interactions. Collectively, our results indicated that targeting PDK1 with DAP inhibited AML cell growth via multiple signaling pathways and suggest that targeting PDK1 may be a promising therapeutic strategy for AMLs.