Study on the function and mechanism of long non-coding RNA DMTF1v4 in the occurrence of colon cancer.

OBJECTIVE: To investigate the expression of long non-coding RNA (lncRNA) DMTF1v4 in colon cancer, and the relationship between its expression and disease occurrence. MATERIALS AND METHODS: Human colon cancer tissues and para-carcinoma tissues were harvested. The expression of lncRNA DMTF1v4 was measured by semi-quantitative PCR. The expression of DMTF1v4 in HT-29 colon cancer cells was downregulated using siRNA, and the effect of its downregulation on cell growth was determined by MTT assay and plate clone assay. The effect of DMTF1v4 downregulation on colon cancer cell migration was determined using a transwell assay and scratch wound assay. The effect of DMTF1v4 on colon cancer cell apoptosis was determined using Annexin V/PI double-staining. The changes in p-ERK, p-JNK, and p-p38 were measured by Western blot. HT-29 cells with downregulated DMTF1v4 expression were used to establish the subcutaneous heterotopic transplantation tumor model in nude mice to study the effect of DMTF1v4 on tumor growth in animals. RESULTS: Compared with para-carcinoma tissue, lncRNA DMTF1v4 in colon cancer tissue was highly expressed (p<0.001). Downregulating lncRNA DMTF1v4 in HT-29 cells showed that lncRNA DMTF1v4 promotes cell proliferation and migration, and suppresses apoptosis (p<0.05). The effect of lncRNA DMTF1v4 on the ERK/MAPK signaling pathway was evaluated. The expression of p-ERK, p-JNK, and p-p38 was increased significantly compared with the control group (p<0.01). The effect of downregulating DMTF1v4 on tumor growth in animals showed that tumor growth in nude mice was decreased, and the expression of apoptosis-related proteins was increased (p<0.01). CONCLUSIONS: The expression of lncRNA DMTF1v4 is elevated in colon cancer tissues; lncRNA DMTF1v4 promotes colon cancer cell proliferation and migration, and inhibits apoptosis by downregulating the expression of p-ERK, p-JNK, and p-p38, thus affecting the progression of colon cancer. This will provide a basis for the development of new clinical treatments for colon cancer.

IMP3 promotes TNBC stem cell property through miRNA-34a regulation.

OBJECTIVE: To explore the expression and function of insulin-like growth factor II (IGFII) mRNA binding protein (IMP3) in the Triple Negative Breast Cancer (TNBC). MATERIALS AND METHODS: According to previously reported gene expression array, we found that IMP3 had significantly higher expression in the CD44+CD24-ESA+ cell cluster, tumor initiating cell or cancer stem cell (CSCs), compared to other tumor cells. Based on the GEO database (GEO accession No. GSE6883), we detected the mRNA levels of IMP 1,2 and 3 by quantitative polymerase chain reaction (q-PCR) in CD44+CD24-ESA+ cell cluster and other breast tumor cell clusters. Besides, we measured IMP3 expression in microsphere of breast cancer, which exerted more significant tumor stem cell properties. The effects of IMP3 on breast cancer cell stem cell properties were studied by RNA interference and overexpression approaches in vitro. Furthermore, we predicted and identified microRNA, which could target and regulate IMP3 from bioinformatics analysis, and verified the interaction by luciferase assays and rescue experiments. RESULTS: Previously reported data showed that IMP3 expression was significantly upregulated in CD44+CD24-ESA+ cell cluster from breast cancer tissues. Besides, we found IMP3 had higher expression in mesenchymal cells rather than epithelial cells, which was also significantly elevated in SUM159 and T49D cell lines cultured as microsphere rather than adherent cells or differentiated cells. CD44+CD24-ESA+ cell cluster proportion was significantly decreased after silencing IMP3 in SUM1315, and its ability to develop into microsphere was significantly inhibited. By re-expressing IMP3 in SUM315, we restored the self-renewal capacity and tumorigenesis potential of SUM315. Through relative predicting website, we found several miRNAs which could regulate IMP3. miR-34a with highest score was chosen for further analysis. Mimicking miR-34a significantly downregulated IMP3 expression and inhibited its ability to develop into microsphere, while overexpressing IMP3 could rescue this process. CONCLUSIONS: IMP3 plays a vital role in maintaining stem cell properties of breast cancer cells, which could be regulated by mir-34a.

Transcriptional regulation of the vitellogenin gene through a fecundity-related single nucleotide polymorphism within a GATA-1 binding motif in the brown planthopper, Nilaparvata lugens.

Identifying the Single Nucleotide Polymorphisms (SNPs) with functions in insect fecundity promises to provide novel insight into genetic mechanisms of adaptation and to aid in effective control of insect populations. We previously identified several SNPs within the vitellogenin (Vg) promoter region between a high-fecundity population (HFP) and a low-fecundity population (LFP) of the brown planthopper, Nilaparvata lugens Stål (Hemiptera: Delphacidae). Here, we found that an A-to-T (HFP allele to LFP allele) transversion at nucleotide -953 upstream of Vg in a Nilaparvata lugens GATA-1 (NlGATA-1) binding motif is associated with the level of Vg transcription. We also characterized NlGATA-1, containing a double CX2 CX17 CX2 C zinc finger, which has been implicated in the activation of Vg gene expression. Knockdown of the NlGATA-1 gene results in a reduced basal level of expression of the Vg gene and fewer offspring of N. lugens in vivo, whereas overexpression of NlGATA-1 in cells increased Vg promoter activity. Moreover, upon cotransfection with NlGATA-1 expression vector, the luciferase activities of Vg reporter vectors with the A allele were significantly higher than those with the T allele. These findings support a mechanism in which a SNP within the promoter of Vg is associated with the level of Vg transcription by altering the binding activity of NlGATA-1 and subsequently affecting fecundity in N. lugens.

A diagnostic algorithm for assessment of liver fibrosis by liver stiffness measurement in patients with chronic hepatitis B.

Steatosis could affect liver stiffness measurement in patients with nonalcoholic fatty liver disease and chronic hepatitis C. In this study, we aimed to investigate the impact of steatosis on liver stiffness in hepatitis B virus (HBV)-infected patients and develop a diagnostic algorithm for prediction of liver fibrosis by liver stiffness based on the controlled attenuation parameter. A total of 488 HBV-infected patients who underwent clinical examination, Fibroscan and liver biopsy were prospectively enrolled. The best liver stiffness measurement (kPa) cut-offs for significant fibrosis (S≥3) and advanced fibrosis (S≥4) were 8.1 and 10.9, respectively. The best controlled attenuation parameter cut-off for severe steatosis (≥30%) was 287 dB/m. Among patients with low-grade fibrosis (S0-S2/S0-S3), mean liver stiffness values were significantly higher in subjects with severe steatosis or controlled attenuation parameter ≥287 dB/m compared with those without. Moreover, in subjects with low-grade fibrosis, a higher rate of false-positive rate was observed in patients with severe steatosis than those in patients without (F0-F2: 28.2% vs 9.7%; F0-F3: 17.0% vs 5.3%), and in patients with CAP≥287 dB/m compared with their counterpart (F0-F2: 23.7% vs 9.2%; F0-F3: 14.1% vs 4.8%). Low-grade fibrosis was accurately identified by γ-glutamyl transpeptidase-to-platelet ratio (GPR) with a cut-off value of 0.17. In patients with GPR<0.17, similar results were observed. The presence of steatosis may lead to overestimation of fibrosis assessed by liver stiffness measurement in patient with chronic hepatitis B. A diagnostic algorithm for assessing fibrosis using liver stiffness was developed by combining both controlled attenuation parameter and GPR values.

A nomogram for predicting prognostic value of inflammatory response biomarkers in decompensated cirrhotic patients without acute-on-chronic liver failure.

BACKGROUND: Inflammation plays a vital role in liver cirrhosis progression and prognosis. AIM: To investigate the prognostic significance of inflammatory response markers in decompensated cirrhotic patients without acute-on-chronic liver failure (ACLF). METHODS: Independent predictors were identified using multivariate Cox model and then assembled into a nomogram to predict survival. Concordance index (C-index) and time-dependent receiver operating characteristics (td-ROC) analysis were adopted to evaluate and compare the performance of nomogram, model for end-stage liver disease (MELD) scores, MELD-Na and Chronic Liver Failure-consortium score for acute decompensated (CLIF-C ADs). RESULTS: A total of 902 decompensated cirrhotic patients with different aetiologies were enrolled, with 6-month, 1-year and 3-year mortality of 18.6%, 24.4% and 34.8%, respectively. The cut-off values for neutrophil-to-lymphocyte ratio (NLR) and lymphocyte-to-monocyte ratio (LMR) determined by X-tile program were 5.7 and 1.1 respectively. Patients with NLR>5.7 or LMR≤1.1 had significantly higher mortality (P < 0.001). Independent factors derived from multivariable Cox analysis of development cohort to predict mortality were age, NLR and LMR (hazard ratio (HR): 1.064, 95% confidence interval (CI): 1.045-1.084, P < 0.001; HR: 1.124, 95%CI: 1.091-1.158, P < 0.001; HR: 0.794, 95%CI: 0.702-0.898, P < 0.001, respectively). The C-indexes of nomogram were higher than that of MELD score, MELD-Na and CLIF-C ADs for predicting survival. The tdROC and decision curves showed that nomogram was superior to MELD score, MELD-Na and CLIF-C ADs. Similar results were observed in validation cohort. CONCLUSION: The proposed nomogram with neutrophil-to-lymphocyte ratio and lymphocyte-to-monocyte ratio resulted in accurate prognostic prediction for decompensated cirrhotic patients without ACLF.

Usefulness of albumin-bilirubin grade for evaluation of long-term prognosis for hepatitis B-related cirrhosis.

Long-term prognosis varies widely among patients with hepatitis B virus (HBV)-related liver cirrhosis. Our study aimed to investigate the applicability of albumin-bilirubin (ALBI), Child-Pugh and model for end-stage liver disease (MELD) scores to the long-term prognosis prediction of HBV-related cirrhosis. Patients diagnosed with HBV-associated cirrhosis from the First Affiliated Hospital of Wenzhou Medical University between January 2010 and December 2015 were enrolled in this study. The patients were followed up every 3 months. The prognostic performance of ALBI in long-term outcome prediction for HBV-related cirrhosis was compared with Child-Pugh and MELD scores using time-dependent receiver operating characteristic curve (tdROC) and decision curve analysis. A total of 806 patients were included in our study with 275 (34.1%) deceased during the follow-up. Multivariate Cox regression analysis showed that ALBI grade was an independent predictor associated with mortality. The tdROC analysis showed that ALBI score (0.787, 0.830 and 0.833) was superior to MELD (0.693, P=.003; 0.717, P<.001; 0.744, P<.001) and Child-Pugh score (0.641, P<.001; 0.649, P<.001; 0.657, P<.001) for predicting 1-year, 2-year and 3-year mortality. Additionally, decision curves also got the similar results. In addition, patients with lower ALBI score had a longer life expectancy, even among patients within the same Child-Pugh class. Thus, ALBI score was effective in predicting the long-term prognosis for patients with HBV-related cirrhosis and more accurate than Child-Pugh and MELD scores.

Effect of FUT3 gene silencing with miRNA on proliferation, invasion and migration abilities of human KATO-III gastric cancer cell line.

This study investigated the effects of FUT3 gene expression inhibition with miRNA on the proliferation, invasion and migration abilities of KATO-III cells. KATO-III cells were transfected with plasmid pcDNA™6.2-GW/EmGFP-FUT3-miR(FUT3-miRNA) and negative control plasmid in mediation of liposome, respectively, using untransfected cells as blank controls. Forty-eight hours after transfection, FUT3 mRNA levels were tested by RT-PCR. Levels of sLeA proteins were assayed by Western blot. The effects of FUT3-miRNA on the proliferation, invasion and migration of KATO-III cells were determined by CCK8 testing and Transwell assays, respectively. Results indicate that the transfection of FUT3-miRNA may down-regulate sLeA protein expression on the surface of KATO-III cells, and significantly inhibit cell proliferation (p<0.05). As compared to the negative and blank control groups, the number of invasion and migration cells in the FUT3-miRNA group decreased significantly (each p<0.05). Experimental results indicate that the miRNA expression vector which targets the FUT3 gene can effectively inhibit the proliferation, migration and invasion abilities of KATO-III cells.

Relationship between Serum Aminotransferase Levels and Metabolic Disorders in Northern China.

BACKGROUND: Increasing evidence suggests an association between elevated serum aminotransferase levels and metabolic disorders (metabolic syndrome, hyperlipemia and diabetes mellitus). However, the significance of relatively low levels of aminotransferases in relation to metabolic disorders has not been fully investigated in the general population. We investigated the association between serum amiontransferase levels and metabolic disorders using data from a survey in Jilin province, China. METHODS: In 2007, a survey was conducted throughout Jilin, China, covering both urban and rural areas. A total of 3835 people, 18 to 79 years old including 1761 men and 2074 women, underwent real-time ultrasonography, blood tests including aspartate aminotransferase and alanine aminotransferase, and had interviews with a structured questionnaire. RESULTS: Serum aminotransferase levels within the normal range were associated with metabolic syndrome independent of age, occupation, cultural and educational level, income, body mass index, waist circumference, smoking, and alcohol intake. Compared with the lowest level (<20 IU/L), the adjusted odds ratios for ALT levels of 20-29, 30-39, 40-49 and >50 IU/L were 1.92, 2.50, 2.97, and 3.52 in men, and 1.38, 1.54, 3.06, and 2.62 in women, respectively. Near-normal serum aminotransferase levels associated with hyperlipemia, NAFLD, DM were also found in the study. CONCLUSIONS: Normal to near-normal serum aminotransferase levels are associated with metabolic disorders. Serum ALT levels of 21-25 IU/L for men, and 17-22 IU/L for women are suggested as cutoff levels that detect metabolic disorders affecting the liver.

Fermentability of the hemicellulose-derived sugars from steam-exploded softwood (douglas fir)

Steam explosion ofDouglas fir wood chips under low-severity conditions (log Ro = 3.08 corresponding to 175 degrees C, 7.5 min, and 4.5% SO2) resulted in the recovery of around 87% of the original hemicellulose component in the water-soluble stream. More than 80% of the recovered hemicellulose was in a monomeric form. As the pretreatment severity increased from 3.08 to 3.76, hemicellulose recovery dropped to 43% of the original hemicellulose found in Douglas fir chips while the concentration of glucose originating from cellulose hydrolysis increased along with the concentration of sugar degradation products such as furfural and hydroxymethylfurfural. Despite containing a higher concentration of hexose monomers (mainly glucose originating from cellulose degradation), the water-soluble fraction prepared under high-severity conditions (log Ro = 3.73 corresponding to 215 degrees C, 2.38 min, and 2.38% SO2) was not readily fermented. Only the two hydrolyzates obtained at low and medium (195 degrees C, 4.5 min, and 4.5% SO2) severities were fermented to ethanol using a spent sulfur liquor adapted strain of Saccharomyces cerevisiae. High ethanol yields were obtained for these two hydrolyzates with 0.44 g of ethanol produced per gram of hexose utilized (86% of theoretical). However, the best results of hemicellulose recovery and fermentability were obtained for the low-severity water-soluble fraction which was fermented significantly faster than the fraction obtained after medium-severity treatment probably because it contained higher amounts of fermentation inhibitors. Copyright 1999 John Wiley & Sons, Inc.