OBJECTIVE

Increased de novo lipogenesis (DNL) contributes to the pathogenesis of nonalcoholic steatohepatitis (NASH). Acetyl-CoA carboxylase catalyzes the rate-limiting step in DNL. We evaluated the safety and efficacy of GS-0976, a small molecule inhibitor of acetyl-CoA carboxylase, in patients with NASH.

METHODS

In an open-label prospective study, patients with NASH (n = 10) received GS-0976 20 mg orally once daily for 12 weeks. NASH was diagnosed based on a proton density fat fraction estimated by magnetic resonance imaging (MRI-PDFF) ≥10% and liver stiffness by magnetic resonance elastography (MRE) ≥2.88 kPa. The contribution from hepatic DNL to plasma palmitate was measured by 14 days of heavy water labeling before and at the end of treatment. We performed the same labelling protocol in an analysis of healthy volunteers who were not given DNL (controls, n = 10). MRI-PDFF and MRE at baseline, and at weeks 4 and 12 of GS-0976 administration, were measured. We analyzed markers of liver injury and serum markers of fibrosis.

RESULTS

The contribution of hepatic DNL to plasma palmitate was significantly greater in patients with NASH compared with controls (43% vs 18%) (P = .003). After 12 weeks administration of GS-0976, the median hepatic DNL was reduced 22% from baseline in patients with NASH (P = .004). Compared with baseline, reductions in MRI-PDFF at week 12 (15.7% vs 9.1% at baseline; P = .006), liver stiffness by MRE (3.4 kPa vs 3.1 kPa at baseline; P = .049), TIMP metallopeptidase inhibitor 1 (275 ng/mL vs 244 ng/mL at baseline; P = .049), and serum level of alanine aminotransferase (101 U/L vs 57 U/L at baseline; P = .23) were consistent with decreased hepatic lipid content and liver injury. At week 12, 7 patients (70%) had a ≥30% decrease in MRI-PDFF.

CONCLUSIONS

In an open-label study, patients with NASH given GS-0976 for 12 weeks had reduced hepatic DNL, steatosis, and markers of liver injury. ClinicalTrials.gov no: NCT02856555.

BACKGROUND

Approximately 30% of fine-needle aspiration (FNA) biopsies of thyroid nodules are indeterminate, nondiagnostic, or suspicious. The purpose of the current study was to determine the accuracy of novel candidate diagnostic markers to distinguish benign from malignant thyroid neoplasms, and to predict the extent of disease.

METHODS

A real-time quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) assay of 6 novel candidate diagnostic and extent of disease marker genes (extracellular matrix protein 1 [ECM1]; transmembrane protease, serine <em>4</em> [TMPRSS<em>4</em>]; angiopoietin 2 [ANGPT2]; <em>TIMP</em> <em>metallopeptidase</em> <em>inhibitor</em> 1 [<em>TIMP</em>1]; ephrin-B2 [EFNB2], and epidermal growth factor receptor [EGFR]) was used in 126 thyroid tissues. To evaluate the performance of the scoring model for the diagnostic markers in combination, the area under the receiver operating characteristic (ROC) curve (AUC) was determined.

RESULTS

The levels of ECM1, TMPRSS<em>4</em>, ANGPT2, and <em>TIMP</em>1 mRNA expression were found to be independent diagnostic markers of malignant thyroid neoplasms. The AUC for the <em>4</em> diagnostic genes in combination was 0.993 with a sensitivity of 100%, a specificity of 9<em>4</em>.6%, a positive predictive value of 96.5%, and a negative predictive value of 100%. In 31 thyroid nodule FNA biopsy samples, the scoring model had a sensitivity of 91.0%, a specificity of 95.0%, a positive predictive value of 92.9%, and a negative predictive value of 92.3%. The multigene assay correctly classified 93% of tumors into the correct risk group (low-risk vs. high-risk) with a sensitivity of 78.9% (true positive in high-risk tumors), specificity of 92% (true negative in low-risk tumors), positive predictive value of 87.5%, and negative predictive value of 92%. In 11 malignant thyroid nodule FNA samples, the extent of disease scoring model correctly identified 3 of <em>4</em> high-risk differentiated thyroid cancers and 7 of 7 low-risk differentiated thyroid cancers.

CONCLUSIONS

This novel multigene assay is an excellent diagnostic and extent of disease marker for differentiated thyroid cancer and would be a helpful adjunct to FNA biopsy of thyroid nodules.

The aim of this study was to investigate the effect of quercetin on hepatic fibrosis, a characteristic response to acute or chronic liver injury. Mice were randomized to bile duct ligation (BDL) or carbon tetrachloride (CCl<em>4</em>) cirrhosis models. Quercetin (100 mg/kg or 200 mg/kg daily) was administered by gavage for 2 or <em>4</em> weeks. Liver tissue and blood samples were collected for histological and molecular analysis. The results of our experiments showed that quercetin reduced BDL or CCl<em>4</em> liver fibrosis, <em>inhibited</em> extracellular matrix formation, and regulated matrix <em>metallopeptidase</em> (MMP)-9 and tissue <em>inhibitor</em> of metalloproteinase (<em>TIMP</em>)-1. Quercetin attenuated liver damage by suppressing the TGF-β1/Smads signaling pathway and activating the PI3K/Akt signaling pathway to inhibit autophagy in BDL- or CCl<em>4</em>- induced liver fibrosis. Quercetin prevented hepatic fibrosis by attenuating hepatic stellate cell activation and reducing autophagy through regulating crosstalk between the TGF-β1/Smads and PI3K/Akt pathways.

Epidemiological studies demonstrate that the incidence and mortality rates of colorectal cancer in women are lower than in men. However, it is unknown if 17β-estradiol (E(2)) treatment is sufficient to inhibit cell proliferation and cell migration in human colon cancer cells. Up-regulation of urokinase plasminogen activator (uPA), tissue plasminogen activator (tPA), and matrix <em>metallopeptidases</em> (MMPs) is reported to associate with the development of cancer cell mobility, metastasis, and subsequent malignant tumor. In the present study, we treated human LoVo colon cancer cells with E(2) to explore whether E(2) down-regulates cell proliferation and migration, and to identify the precise molecular and cellular mechanisms behind the down-regulatory responses. Here, we found that E(2) treatment decreased cell proliferation and cell cycle-regulating factors such as cyclin A, cyclin D1 and cyclin E. At the same time, E(2) significantly <em>inhibited</em> cell migration and migration-related factors such as uPA, tPA, MMP-2, and MMP-9. However, E(2) treatment showed no effects on upregulating expression of plasminogen activator <em>inhibitor</em>-1 (PAI-1), tissue <em>inhibitor</em> of metalloproteinase-1, -2, -3, and -<em>4</em> (<em>TIMP</em>-1, -2, -3, and -<em>4</em>). After administration of <em>inhibitors</em> including QNZ (NFκB <em>inhibitor</em>), LY29<em>4</em>002 (Akt activation <em>inhibitor</em>), U0126 (ERK1/2 <em>inhibitor</em>), SB203580 (p38 MAPK <em>inhibitor</em>) or SP600125 (JNK1/2 <em>inhibitor</em>), E(2) -downregulated cell migration and expression of MMP-2 and MMP-9 in LoVo cells is markedly <em>inhibited</em> only by p38 MAPK <em>inhibitors</em>, SB203580. Application of specific target gene siRNA (ERα, ERβ, p38α, and p38β) to LoVo cells further confirmed that p38 MAPK mediates E(2) /ERs <em>inhibition</em> of MMP-2 and -9 expression and cell motility in LoVo cells. Collectively, these results suggest that E(2) treatment down-regulates cell proliferation by modulating the expression of cyclin A, cyclin D1 and cyclin E. E(2) treatment simultaneously impaired cell migration by inhibiting the expression of uPA, tPA, MMP-2, and MMP-9 through E(2) /ERs - p38α MAPK signaling pathway in human LoVo colon cancer cells.

Upregulation of urokinase plasminogen activator (uPA), tissue plasminogen activator (tPA), and matrix <em>metallopeptidases</em> (MMPs) is associated with the development of myocardial infarction (MI), dilated cardiomyopathy, cardiac fibrosis, and heart failure (HF). Evidences suggest that lipopolysaccharide (LPS) participates in the inflammatory response in the cardiovascular system; however, it is unknown if LPS is sufficient to upregulate expressions and/or activity of uPA, tPA, MMP-2, and MMP-9 in myocardial cells. In this study, we treated H9c2 cardiomyoblasts with LPS to explore whether LPS upregulates uPA, tPA, MMP-2, and MMP-9, and further to identify the precise molecular and cellular mechanisms behind this upregulatory responses. Here, we show that LPS challenge increased the protein levels of uPA, MMP-2 and MMP-9, and induced the activity of MMP-2 and MMP-9 in H9c2 cardiomyoblasts. However, LPS showed no effects on the expression of tissue <em>inhibitor</em> of metalloproteinase-1, -2, -3, and -<em>4</em> (<em>TIMP</em>-1, -2, -3, and -<em>4</em>). After administration of <em>inhibitors</em> including U0126 (ERK1/2 <em>inhibitor</em>), SB203580 (p38 MAPK <em>inhibitor</em>), SP600125 (JNK1/2 <em>inhibitor</em>), CsA (calcineurin <em>inhibitor</em>), and QNZ (NFkappaB <em>inhibitor</em>), the LPS-upregulated expression and/or activity of uPA, MMP-2, and MMP-9 in H9c2 cardiomyoblasts are markedly <em>inhibited</em> only by ERK1/2 <em>inhibitors</em>, U0126. Collectively, these results suggest that LPS upregulates the expression and/or activity of uPA, MMP-2, and MMP-9 through ERK1/2 signaling pathway in H9c2 cardiomyoblasts. Our findings further provide a link between the LPS-induced cardiac dysfunction and the ERK1/2 signaling pathway that mediates the upregulation of uPA, MMP-2 and MMP-9.

The neuropeptide S (NPS)-NPS receptor 1 (NPSR1) pathway has recently been implicated in the pathogenesis of asthma. The purpose of this study was to identify downstream gene targets regulated by NPSR1 upon NPS stimulation. A total of 10<em>4</em> genes were found significantly up-regulated and <em>4</em>2 down-regulated by microarray analysis 6 h after NPS administration. By Gene Ontology enrichment analysis, the categories 'cell proliferation', 'morphogenesis' and 'immune response' were among the most altered. A TMM microarray database comparison suggested a common co-regulated pathway, which includes JUN/FOS oncogene homologs, early growth response genes, nuclear receptor subfamily <em>4</em> members and dual specificity phosphatases. The expression of four up-regulated genes, matrix <em>metallopeptidase</em> 10 (MMP10), INHBA (activin A), interleukin 8 (IL8) and EPH receptor A2 (EPHA2), exhibited a significant NPS dose-response relationship as confirmed by quantitative reverse-transcriptase-PCR and for MMP10 by immunoassay. Immunohistochemical analyses revealed that MMP10 and <em>TIMP</em> <em>metallopeptidase</em> <em>inhibitor</em> 3 (<em>TIMP</em>3) were both strongly expressed in bronchial epithelium, and macrophages and eosinophils expressed MMP10 in asthmatic sputum samples. Because remodeling of airway epithelium is a feature of chronic asthma, the up-regulation of MMP10 and <em>TIMP</em>3 by NPS-NPSR1 signaling may be of relevance in the pathogenesis of asthma.

BACKGROUND

Studies have reported that the integrated analysis of microRNA (miRNA)-messenger RNA (mRNA) expression is valuable in exploring gene regulation systemically.

OBJECTIVE

The objectives of this study were to identify miRNAs and genes involved in atrial fibrillation and to explore the mechanisms underlying atrial fibrosis.

METHODS

We used microarrays to compare the differences in both miRNA and mRNA expression profiles in the left atrial appendage of patients with nonvalvular paroxysmal atrial fibrillation and healthy controls. Furthermore, the quantitative real-time polymerase chain reaction was used to confirm the reliability of the microarray data, prediction of the adopted databases, and Ingenuity Pathway Analysis of miRNA-mRNA expression in order to identify the miRNA target genes, examine the functions and pathways in which the target genes are involved, and construct an miRNA-target gene regulatory network. We further investigated the roles of miRNA-1<em>4</em>6b-5p in the mechanisms of atrial fibrosis.

RESULTS

We identified 10 differentially expressed miRNAs and 62<em>4</em> differentially expressed mRNAs, among which only 1 miRNA-target gene pair miR-1<em>4</em>6b-5p and tissue <em>inhibitor</em> of metalloproteinase <em>4</em> (<em>TIMP</em>-<em>4</em>) were constructed. The validated results revealed that miR-1<em>4</em>6b-5p, matrix <em>metallopeptidase</em> 9, and collagen content were upregulated whereas <em>TIMP</em>-<em>4</em> was downregulated in patients with atrial fibrillation. After the transfection of miR-1<em>4</em>6b-5p into cardiac fibroblasts, <em>TIMP</em>-<em>4</em> expression was markedly reduced and collagen content was increased. Moreover, luciferase results confirmed that <em>TIMP</em>-<em>4</em> was a target of miR-1<em>4</em>6b-5p.

CONCLUSIONS

The identified miRNA and mRNA may represent a potentially novel molecular regulatory network, which may provide a better understanding of the molecular basis of remodeling in atrial fibrillation. miR-1<em>4</em>6b-5p probably acts as an intracellular mediator in the maladaptive remodeling in atrial fibrosis in atrial fibrillation.

Primary carcinomas of the small intestine are rare and the mechanism of their pathogenesis is poorly understood. Patients with familial adenomatous polyposis (FAP) have a high risk of developing duodenal carcinomas. The aim of this study is to gain more insight into the development of duodenal carcinomas. Therefore, five FAP-related duodenal carcinomas were characterized for chromosomal and methylation alterations, which were compared to those observed in sporadic duodenal carcinomas. Comparative genomic hybridization (CGH) and methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) was performed in 10 primary sporadic and five primary FAP-related duodenal carcinomas. In the FAP-related carcinomas, frequent gains were observed on chromosomes 8, 17 and 19, whereas in sporadic carcinomas they occurred on chromosomes 8, 12, 13 and 20. In 60% of the sporadic carcinomas, gains in the regions of chromosome 12 were observed which were absent in the FAP-related carcinomas (P=0.0<em>4</em>). Hypermethylation was observed in the immunoglobulin superfamily genes member <em>4</em> (IGSF<em>4</em>), <em>TIMP</em> <em>metallopeptidase</em> <em>inhibitor</em> 3 (<em>TIMP</em>3), Estrogen receptor 1 (ESR1), adenomatous polyposis coli (APC), H-cadherin (CDH13) and paired box gene 6 (PAX6) genes. Hypermethylation of PAX6 was only observed in FAP-related carcinomas (3/5) and not in sporadic carcinomas (P=0.02). In conclusion, in contrast to sporadic duodenal carcinomas, gains on chromosome 12 were not observed in duodenal carcinomas of patients with FAP. Identification of the genes in these regions of chromosome 12 could lead to a better understanding of the carcinogenesis pathways leading to sporadic and FAP-related duodenal carcinomas. Furthermore, hypermethylation seems to be a general feature of both FAP-related duodenal carcinomas as well as sporadic duodenal carcinomas with the exception of the PAX6 gene, which is methylated only in FAP-related carcinomas.

BACKGROUND

Epidemiological studies demonstrate that the incidence and mortality rates of colorectal cancer in women are lower than in men. However, it is unknown if 17β-estradiol treatment is sufficient to inhibit prostaglandin E2 (PGE2)-induced cellular motility in human colon cancer cells.

METHODS

We analyzed the protein expression of urokinase plasminogen activator (uPA), tissue plasminogen activator (tPA), matrix metallopeptidases (MMPs), plasminogen activator inhibitor-1 (PAI-1) and tissue inhibitor of metalloproteinases (TIMPs), and the cellular motility in PGE2-stimulated human LoVo cells. 17β-Estradiol and the inhibitors including LY294002 (Akt activation inhibitor), U0126 (ERK1/2 inhibitor), SB203580 (p38 MAPK inhibitor), SP600125 (JNK1/2 inhibitor), QNZ (NFκB inhibitor) and ICI 182 780 were further used to explore the inhibitory effects of 17β-estradiol on PGE2-induced LoVo cell motility. Student's t-test was used to analyze the difference between the two groups.

RESULTS

Upregulation of urokinase plasminogen activator (uPA), tissue plasminogen activator (tPA) and matrix metallopeptidases (MMPs) is reported to associate with the development of cancer cell mobility, metastasis, and subsequent malignant tumor. After administration of inhibitors including LY294002, U0126, SB203580, SP600125 or QNZ, we found that PGE2 treatment up-regulated uPA and MMP-9 expression via JNK1/2 signaling pathway, thus promoting cellular motility in human LoVo cancer cells. However, PGE2 treatment showed no effects on regulating expression of tPA, MMP-2, plasminogen activator inhibitor-1 (PAI-1), tissue inhibitor of metalloproteinase-1, -2, -3 and -4 (TIMP-1, -2, -3 and -4). We further observed that 17β-estradiol treatment inhibited PGE2-induced uPA, MMP-9 and cellular motility by suppressing activation of JNK1/2 in human LoVo cancer cells.

CONCLUSIONS

Collectively, these results suggest that 17β-estradiol treatment significantly inhibits PGE2-induced motility of human LoVo colon cancer cells.

Pre-eclampsia (PE) is a leading cause of maternal and perinatal morbidity and mortality but the exact underlying mechanisms of PE pathogenesis remain elusive. Accumulated data suggested that the long noncoding RNAs (lncRNAs) play important roles in the pathogenesis of PE. The present study identified the changes of lncRNA Linc00261 in PE and its effects on trophoblasts invasion and migration. Our results showed that the expression of Linc00261 was upregulated in placental tissues of PE women compared with those of healthy pregnant women. Overexpression of Linc00261 suppressed cell invasion and migration, induced cell apoptosis, and caused cell-cycle arrest at G₀ /G₁ phase of HTR-8/SVneo cells; while knockdown of Linc00261 had the opposite effects on the HTR-8/SVneo cells. Mechanistic studies showed Linc00261 functioned as a competing endogenous RNA for miR-558 in HTR-8/SVneo cells, and miR-558 was negatively regulated by Linc00261. The expression level of miR-558 in the PE group was significantly lower than the control group, and the expression level of Linc00261 was negatively correlated with the expression level of miR-558 in the placental tissues of women with PE. Furthermore, miR-558 was found to negatively regulate the expression of <em>TIMP</em> <em>metallopeptidase</em> <em>inhibitor</em> <em>4</em> (<em>TIMP</em><em>4</em>) via targeting the 3' untranslated region in the HTR-8/SVneo cells. Overexpression of miR-558 increased HTR-8/SVneo cell invasion and migration, which was attenuated by <em>TIMP</em><em>4</em> overexpression. More importantly, both overexpression of miR-558 and knockdown of <em>TIMP</em><em>4</em> partially reversed the suppressive effects of Linc00261 overexpression on cell invasion and migration of HTR-8/SVneo cells. Collectively, our results for the first time showed the upregulation of Linc00261 in the placental tissues of severe PE patients. The mechanistic results indicated that Linc00261 exerted the suppressive effects on the trophoblast invasion and migration via targeting miR-558/<em>TIMP</em><em>4</em> axis, which may involve in the pathogenesis of PE.

BACKGROUND

Well-annotated clinical samples are valuable resources for biomarker discovery and validation. Multiplex and integrated methods that simultaneously measure multiple analytes and generate integrated information about these analytes from a single measurement are desirable because these methods help conserve precious samples. We developed a magnetic bead-based system for multiplex and integrated glycoprotein quantification by immunoassays and glycan detection by lectin immunosorbent assays (LISAs).

METHODS

Magnetic beads coupled with antibodies were used for capturing proteins of interest. Biotinylated antibodies in combination with streptavidin-labeled phycoerythrin were used for protein quantification. In the LISAs, biotinylated detection antibodies were replaced by biotinylated lectins for glycan detection.

RESULTS

Using tissue <em>inhibitor</em> of <em>metallopeptidase</em> 1 (<em>TIMP</em>-1), tissue plasminogen activator, membrane metallo-endopeptidase, and dipeptidyl peptidase-IV (DPP-<em>4</em>) as models, we found that the multiplex integrated system was comparable to single immunoassays in protein quantification and LISAs in glycan detection. The merits of this system were demonstrated when applied to well-annotated prostate cancer tissues for validation of biomarkers in aggressive prostate cancer. Because of the system's multiplex ability, we used only 300 ng of tissue protein for the integrated detection of glycans in these proteins. Fucosylated <em>TIMP</em>-1 and DPP-<em>4</em> offered improved performance over the proteins in distinguishing aggressive and nonaggressive prostate cancer.

CONCLUSIONS

The multiplex and integrated system conserves samples and is a useful tool for validation of glycoproteins and their glycoforms as biomarkers.

Colorectal cancer (CRC) is a life-threatening disease with a poor prognosis. Therefore, it is crucial to identify molecular prognostic biomarkers for CRC. The present study aimed to identify potential key genes that could be used to predict the prognosis of patients with CRC. Three CRC microarray datasets (GSE20916, GSE73360 and GSE<em>4</em><em>4</em>861) were downloaded from the Gene Expression Omnibus (GEO) database, and one dataset was obtained from The Cancer Genome Atlas (TCGA) database. The three GEO datasets were analyzed to detect differentially expressed genes (DEGs) using the BRB-ArrayTools software. Functional and pathway enrichment analyses of these DEGs were performed using the Database for Annotation, Visualization and Integrated Discovery tool. A protein-protein interaction (PPI) network of DEGs was constructed, hub genes were extracted, and modules of the PPI network were analyzed. To investigate the prognostic values of the hub genes in CRC, data from the CRC datasets of TCGA were used to perform the survival analyses based on the sample splitting method and Cox regression model. Correlation among the hub genes was evaluated using Spearman's correlation analysis. In the three GEO datasets, a total of 105 common DEGs were identified, including 51 down- and 5<em>4</em> up-regulated genes in CRC compared with normal colorectal tissues. A PPI network consisting of 100 DEGs and 551 edges was constructed, and <em>4</em><em>4</em> nodes were identified as hub genes. Among these <em>4</em><em>4</em> genes, the four hub genes <em>TIMP</em> <em>metallopeptidase</em> <em>inhibitor</em> 1 (<em>TIMP</em>1), solute carrier family <em>4</em> member <em>4</em> (SLC<em>4</em>A<em>4</em>), aldo-keto reductase family 1 member B10 (AKR1B10) and ATP binding cassette subfamily E member 1 (ABCE1) were associated with overall survival (OS) in patients with CRC. Three significant modules were extracted from the PPI network. The hub gene <em>TIMP</em>1 was present in Module 1, ABCE1 was involved in Module 2 and SLC<em>4</em>A<em>4</em> was identified in Module 3. Univariate analysis revealed that <em>TIMP</em>1, SLC<em>4</em>A<em>4</em>, AKR1B10 and ABCE1 were associated with the OS of patients with CRC. Multivariate analysis demonstrated that SLC<em>4</em>A<em>4</em> may be an independent prognostic factor associated with OS. Furthermore, the results from correlation analysis revealed that there was no correlation between <em>TIMP</em>1, SLC<em>4</em>A<em>4</em> and ABCE1, whereas AKR1B10 was positively correlated with SLC<em>4</em>A<em>4</em>. In conclusion, the four key genes <em>TIMP</em>1, SLC<em>4</em>A<em>4</em>, AKR1B10 and ABCE1 associated with the OS of patients with CRC were identified by integrated bioinformatics analysis. These key genes may be used as prognostic biomarkers to predict the survival of patients with CRC, and may therefore represent novel therapeutic targets for CRC.

<AbstractText>The effect of air pollution-related particulate matter (PM) on epithelial barrier function and tight junction (TJ) expression in human nasal mucosa has not been studied to date. This study therefore aimed to assess the direct impact of PM with an aerodynamic diameter less than 2.5 μ (PM2.5) on the barrier function and TJ molecular expression of human nasal epithelial cells.</AbstractText><AbstractText>Air-liquid interface cultures were established with epithelial cells derived from noninflammatory nasal mucosal tissue collected from patients undergoing paranasal sinus surgery. Confluent cultures were exposed to 50 or 100 μg/mL PM2.5 for up to 72 hours, and assessed for 1) epithelial barrier integrity as measured by transepithelial resistance (TER) and permeability of fluorescein isothiocyanate (FITC) <em>4</em> kDa; 2) expression of TJs using real-time quantitative polymerase chain reaction and immunofluorescence staining, and 3) proinflammatory cytokines by luminometric bead array or enzyme-linked immunosorbent assay.</AbstractText><AbstractText>Compared to control medium, 50 and/or 100 μg/mL PM2.5-treatment 1) significantly decreased TER and increased FITC permeability, which could not be restored by budesonide pretreatment; 2) significantly decreased the expression of claudin-1 messenger RNA, claudin-1, occludin and ZO-1 protein; and 3) significantly increased production of the cytokines interleukin-8, <em>TIMP</em> <em>metallopeptidase</em> <em>inhibitor</em> 1 and thymic stromal lymphopoietin.</AbstractText><AbstractText>Exposure to PM2.5 may lead to loss of barrier function in human nasal epithelium through decreased expression of TJ proteins and increased release of proinflammatory cytokines. These results suggest an important mechanism of susceptibility to rhinitis and rhinosinusitis in highly PM2.5-polluted areas.</AbstractText>

BACKGROUND

This study examined whether genetic variants of matrix metallopeptidases (MMPs) and their tissue inhibitors (TIMPs) were associated with angiographic coronary plaque progression (PP) in type 2 diabetic and non-diabetic patients.

METHODS

Four hundred and ninety-nine patients were grouped, who underwent coronary angiography and received repeat examinations after 1-y follow-up. Twelve functional polymorphisms of MMPs and TIMPs were characterized.

RESULTS

Genotype distribution and allele frequency of -1612 5A/6A MMP-3 and 3'UTR C/T TIMP-4 differed between patients with PP and those without in both diabetic and non-diabetic groups after Bonferroni's correction (all P<0.0041667, except for allele frequency of MMP-3 [P=0.007] and genotype/allele frequency of TIMP-4 [P=0.04 and P=0.016, respectively] in diabetes). MMP-3 and TIMP-4 polymorphisms were associated with changes in percent diameter stenosis and minimal lumen diameter in diabetic patients, and changes in cumulative coronary obstruction in both diabetic and non-diabetic patients (all P<0.05). Multivariable regression analysis revealed that hypertension, low HDL-C and genotypes of MMP-3 and TIMP-4 were independent determinants of PP in the whole patients, with these 2 genetic factors being associated with PP in diabetic and non-diabetic subgroups.

CONCLUSIONS

This study demonstrated that MMP-3 and TIMP-4 polymorphisms affect angiographic coronary PP in type 2 diabetic and non-diabetic patients.

Chronic rhinosinusitis (CRS) is characterized by a dysfunctional host-environment interaction at the nasal mucosa. Contributions of host susceptibility factors such as atopy and aspirin sensitivity to CRS pathophysiology are well established. However, clinical studies on the effects of environmental factors are limited. This study investigates the histological and immunological effects of allergen exposure duration in animal models.

Animal study.

A murine model for CRS with nasal polypoid lesions was induced by instilling ovalbumin/staphylococcal enterotoxin B (SEB) into murine nasal cavities for 12 (short term) or 2<em>4</em> weeks (long term). Histopathological changes were observed. Interleukin (IL)-<em>4</em>, IL-17A, IL-10, and interferon (INF)-γ levels from nasal lavage fluid were measured using enzyme-linked immunosorbent assay. Gene expressions of IL-25, thymic stromal lymphopoietin (TSLP), IL-<em>4</em>, IL-5, INF-γ, C-C motif chemokine ligand (CCL)-11, CCL-2<em>4</em>, C-X-C motif chemokine ligand (CXCL)-1, CXCL-2, intercellular adhesion molecule 1, vascular cell adhesion molecule 1, matrix <em>metallopeptidase</em> (MMP)-7, and tissue <em>inhibitor</em> of metalloproteinase (<em>TIMP</em>)-3 were analyzed from the nasal mucosa.

Long-term CRS models exhibited increased polypoid lesions, edematous mucosal thickness, and eosinophil infiltration compared with short-term models and showed a higher IL-10 level but lower IFN-γ and IL-17A protein levels. Moreover, CCL-2<em>4</em> and MMP-7 gene expressions increased whereas <em>TIMP</em>-3 expression decreased in long-term models compared to controls and short-term models. IL-25 and TSLP expressions were upregulated at mRNA and protein levels in short-term and long-term CRS models, respectively. Furthermore, TSLP mRNA expression was positively associated with IL-5 (r = 0.875<em>4</em>) and inversely correlated to IFN-γ (r = -0.7212) in CRS models.

Prolonged allergen exposure in ovalbumin/SEB-induced CRS models maintains Th2 inflammation and reduces Th1 inflammation, which was associated with upregulation of TSLP.

NA Laryngoscope, 126:E265-E272, 2016.

OBJECTIVE

The aim of this study was to measure the level of nerve growth factor (NGF) in bronchial specimens from humans and to determine whether it correlated with not only clinical characteristics of asthma such as percent eosinophils, Th2 cytokine levels, and pulmonary function, but also metallopeptidase-9 (MMP-9) and tissue inhibitor of metalloproteinases-1 (TIMP-1).

METHODS

Fifty-three people participated; 42 had asthma. The participants underwent bronchoscopy and the specimens were analyzed. The participants' clinical data including pulmonary function tests were reviewed.

RESULTS

Bronchoalveolar lavage fluid (BALF) from patients with asthma had a significantly higher level of NGF compared with that from participants without asthma. NGF level showed a positive correlation with the percentage of eosinophils in both BALF and serum. The concentration of NGF did not correlate with that of Th2 cytokines interleukin (IL)-4, IL-5, and IL-13 in BALF or parameters of pulmonary function including degree of airway hyperresponsiveness (ARH). The levels of MMP-9 and TIMP-1 in BALF were higher in asthma patients than in participants without asthma. The levels of NGF correlated with TIMP-1 levels but not with MMP-9 in the whole participants.

CONCLUSIONS

This study shows that NGF correlates with levels of eosinophils, a major effector cell in asthma. The high expression of NGF and TIMP-1 in asthma patients and the moderate correlation between NGF and TIMP-1 in the entire group of asthma subjects suggest a possible association between NGF and TIMP-1, which may influence asthma pathogenesis.

The aim of the present study was to investigate the anticancer activities of Nelumbo nucifera (Ba lotus) stamen ethanol crude extract (BLSEE) in human colon carcinoma HCT-116 cells. MTT assay, flow cytometry analysis and reverse transcription-polymerase chain reaction assay were employed to investigate the anticancer mechanisms of BLSEE (100, 200 and <em>4</em>00 µg/ml) in HCT-116 cells. BLSEE reduced HCT-116 cell proliferation in a dose-dependent manner. BLSEE treatment also significantly increased the sub-G1 population in HCT-116 cells (P=0.0020 at <em>4</em>00 µg/ml), as shown by flow cytometry assay. Following treatment with BLSEE, the mRNA levels of the apoptosis-associated factors Fas, Fas ligand, tumor necrosis factor-related apoptosis-inducing ligand, death receptor <em>4</em> (DR<em>4</em>), death receptor 5 (DR5), caspases 3, 8 and 9, and B-cell lymphoma-2 (Bcl-2) associated X protein were increased, and the expression of anti-apoptotic Bcl-2 and Bcl-extra large was decreased in HCT-116 cells. The mRNA levels of matrix metalloproteinase (MMP)-2, MMP-9, <em>TIMP</em> <em>metallopeptidase</em> <em>inhibitor</em> 1 and <em>TIMP</em> <em>metallopeptidase</em> <em>inhibitor</em> 2 were also regulated by BLSEE treatment. In addition, BLSEE was able to modulate the expression of inflammation-associated nuclear factor-κB, <em>inhibitory</em> κBα, inducible nitric oxide synthase and cyclooxygenase 2 in HCT-116 cells. The present study clearly indicated the cytotoxicity of BLSEE in HCT-116 cells through induced cellular apoptosis. These results also suggested the BLSEE may be a powerful agent against colon cancer cells.

Obesity is marked by deposition of collagen I in adipose tissue. Toll like receptor (TLR)2 is involved in lipid metabolism, however the association between TLR2 and collagen I remains unclear. The present study was designed to investigate the effect of TLR2 knockout on collagen I in adipose tissue in obese mice. TLR2 knockout and C57BL/6J mice (aged <em>4</em> weeks) were fed normal chow or a high‑fat‑diet for 16 weeks. Compared with adipose tissue from lean controls, that from C57BL/6J mice fed a high‑fat diet had increased levels of collagen I, <em>TIMP</em>1 and TGFβ1 and lower levels of MMP1. However, adipose tissue from TLR2 knockout mice fed a high‑fat diet revealed decreased levels of collagen I, <em>TIMP</em> <em>metallopeptidase</em> <em>inhibitor</em> (<em>TIMP</em>)1, and transforming growth factor (TGF)β1, in addition to increased levels of matrix <em>metallopeptidase</em> (MMP)1. These findings suggest that, in the adipose tissue of obese mice, TLR2 is involved in the metabolism of collagen I and may exhibit a role in the metabolism of MMP1, <em>TIMP</em>1 and TGFβ1.

Chronic histiocytic intervillositis of the placenta (CHI) is a rare and potentially recurrent disease. Characteristically it shows accumulation of CD68+ cells in the intervillous space but no destructive tissue infiltration. An immunopathological background is likely but it is unknown what attracts circulating monocytes to the placenta.

METHODS

We analysed the expression profile of 102 inflammation- and angiogenesis-associated genes with real-time reverse transcriptase-polymerase chain reaction (RT-PCR) in 16 placentas: CHI (n = 5) and, as controls, villitis of unknown aetiology (VUE, n = <em>4</em>) and normal placenta (n = 7).

RESULTS

Compared to controls, CHI had significantly higher levels of matrix metallopeptidase 9 (MMP9) and transforming growth factor, beta receptor 1 (TGFBR1). MMP1<em>4</em> was lower in VUE than CHI (p < 0.05) and controls (not significant). Chemokine (C-X-C motif) ligand 9 (CXCL9), CXCL12, chemokine (C-C motif) ligand 5 (CCL5) and TIMP metallopeptidase inhibitor 1 (TIMP1) were significantly higher in VUE versus controls but not deregulated in CHI. The expression profile could not clearly discriminate CHI from VUE or controls but a tendency for grouping of massive CHI was found. Angiogenesis-associated factors were not deregulated in CHI.

CONCLUSIONS

The discrepancy of massive histiocytic accumulation and the lack of striking up-regulation of cytokines might be the basis of the non-destructive behaviour of the histiocytes in CHI.

Asthma is characterized by airway inflammatory infiltration, which leads to airway remodeling and airway hyperreactivity. Coleus forskohlii (CFK) has been used to treat asthma, however, the mechanism involved is not clear. To explore the antiasthma mechanism of extracts of Coleus forskohlii (ECFK), guinea pigs were administered with a spray of phosphoric acid histamine, and rats were sensitized with ovalbumin (OVA). Hematoxylin and eosin staining (H&E) were used to evaluate pathological changes in lung tissue. Enzyme-linked immunosorbent assay (ELISA) was used to determine cytokine levels in serum and bronchoalveolar lavage fluid (BALF). Immunohistochemistry and Western blot analysis were used to assess the expression of intercellular cell adhesion molecule-1 (ICAM-1), phosphorylation of p65 (p-p65), matrix <em>metallopeptidase</em> 9 (MMP-9), and tissue <em>inhibitor</em> of metalloproteinase 1 (<em>TIMP</em>-1). After ECFK treatment, the asthma incubation period of guinea pigs was significantly prolonged. The H&E results showed that the number of eosinophils in the 12.8 g/kg ECFK group was significantly lower when compared with the control group. Moreover, ELISA results demonstrated that interleukin (IL)-<em>4</em>, IL-5, and IL-17 in serum and BALF were significantly decreased, and interferon-γ (IFN-γ) and IL-10 were increased after ECFK treatment. In addition, ECFK treatment resulted in downregulation of ICAM-1, p-p65, MMP-9, and <em>TIMP</em>-1 in lung tissue after being sensitized by OVA. In conclusion, our findings indicated that ECFK significantly alleviated OVA-induced inflammatory infiltration and airway remodeling in asthma. This study laid a theoretical foundation for the clinical use of ECFK.

To identify differences in gene expression profiles of infected cells between thyroid carcinoma (C), thyroid adenoma (A) and normal thyroid (N) epithelial cells, differentially expressed genes were identified using three pairwise comparisons with the GEO2R online tool. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were used to classify them at the functional level. The most significant cluster in the N vs. A pairwise comparison had four hub genes: Insulin-like growth factor 2, Von Willebrand factor (VWF), multimerin 1 (MMRN1) and complement factor D (CFD). In N vs. C, the most significant cluster had 19 genes: IGF2, early growth response 2, transcription factor 3, KIT proto‑oncogene receptor tyrosine kinase, SMAD family member 9, MLLT3 super elongation complex subunit, runt related transcription factor 1, CFD, actinin α 1, SWI/SNF related matrix associated actin dependent regulator of chromatin subfamily a member <em>4</em>, JunD proto‑oncogene AP‑1 transcription factor subunit, serum response factor (SRF), FosB proto‑oncogene, AP‑1 transcription factor subunit, connective tissue growth factor (CTGF), SRC proto‑oncogene, non‑receptor tyrosine kinase, MMRN1, SRY‑box 9, early growth response 3 and ETS variant <em>4</em>. In A vs. C, the most significant cluster had 1<em>4</em> genes: BCL2-like 1, galectin 3, MCL1 BCL2 family apoptosis regulator, DNA damage inducible transcript 3, BCL2 apoptosis regulator, CTGF, matrix <em>metallopeptidase</em> 7, early growth response 1, kinase insert domain receptor, <em>TIMP</em> <em>metallopeptidase</em> <em>inhibitor</em> 1, apolipoprotein E, VWF, cyclin D1 and placental growth factor. Histological evidence was presented to confirm the makeup of the hubs prior to logistic regression analysis to differentiate benign and malignant neoplasms. The results of the present study may aid in the search for novel potential biomarkers for the differential diagnosis, prognosis and development of drug targets of thyroid neoplasm.

<i>Lactobacillus paracasei</i> K5 is a lactic acid bacteria (LAB) strain that has been isolated from dairy products. Previous studies have established its probiotic potential in a series of in vitro tests, including molecular characterization, safety profiling, and tolerability of the gastrointestinal tract conditions. To characterize its beneficial actions on the host, we have shown previously that <i>L. paracasei</i> K5 adheres to Caco-2 cells and exerts anti-proliferative effects through the induction of apoptosis. In the present study, we focused on the immunomodulatory potential of this strain. We employed the dorsal-air-pouch mouse model of inflammation and recorded an eight-fold increase in the recruitment of immune cells in mice treated with the probiotic strain, compared to the control group. Analysis of the exudates revealed significant changes in the expression of pro-inflammatory mediators on site. Treatment of Caco-2 cells with <i>L. paracasei</i> K5 induced significant upregulation of cytokines interleukin-1α (IL-1α), ΙL-1β, IL-6, tumor necrosis factor-alpha (TNF-α), the chemokine C-X-C motif ligand 2 (CXCL2), and the inflammation markers soluble intercellular adhesion molecule (sICAM) and <em>metallopeptidase</em> <em>inhibitor</em>-1 (<em>TIMP</em>-1). Transient induction of the Toll-like receptors (TLRs) 2, <em>4</em>, 6, and 9 expression levels was recorded by real-time PCR analysis. These results highlight the immunomodulatory potential of this strain and further support its probiotic character.

We previously have reported that chondrocyte-derived extracellular matrix (CDECM) suppresses the growth of pterygium in athymic nude mice. The aim of this study is to demonstrate the effect of CDECM on the pterygium epithelial cells and molecular signaling pathways in human primary pterygium epithelial cells (hPECs).

Human conjunctival epithelial cells (hConECs) were used for identification of the effect of CDECM on normal conjunctiva. The effects of CDECM on proliferation were measured with the 3-(<em>4</em>,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(<em>4</em>-sulfenyl)-2H-tetrazolium (MTS) assay. Cell migration was evaluated according to the scratch wound closure assay and the Transwell invasion assay. Pterygium-related angiogenesis, inflammation, and extracellular matrix remodeling were analyzed with immunoblot and enzyme-linked immunosorbent assay (ELISA). The level of oxidative stress was detected with 2',7'-dichlorofluorescein diacetate (DCFH-DA). Protein kinase signaling was also analyzed with immunoblot.

CDECM did not show cytotoxicity until 1 mg/ml in the hConECs and hPECs. Cell migration and invasion were markedly reduced by treatment of 1 mg/ml CDECM in the hPECs to 3<em>4</em>% of the control, but not in the hConECs. CDECM significantly downregulated matrix metallopeptidase 9 (MMP-9) and fibronectin and upregulated tissue inhibitor of metalloprotease 1 (TIMP-1) and -2 in the hPECs. Angiogenic factors, such as vascular endothelial growth factor (VEGF), antivascular cellular adhesion molecule 1 (VCAM-1), and cluster of differentiation 31 (CD31), and proinflammatory factors, including tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (Cox2), interleukin 6 (IL-6), and prostaglandin E2 (PGE2), were dramatically reduced by CDECM in the hPECs. Furthermore, CDECM significantly inhibited the generation of intracellular reactive oxygen species and the expression of NADPH oxidase subunits, Nox2 and p<em>4</em>7phox. CDECM induced nuclear factor erythroid-2 related factor 2 (Nrf2) mediated-antioxidant enzyme heme oxygenase-1 (HO-1). CDECM also suppressed nuclear factor-kappa B (NF-κB) activation and the phosphorylation of p38 mitogen-activated protein kinase (MAPK), protein kinase C alpha (PKCα), and PKCθ.

CDECM was markedly effective in pathogenesis of hPECs. CDECM-suppressed migration of hPECs resulted from the inhibition of NF-κB activation and the improvement of Nrf2 induction by blocking the p38 MAPK and PKC signaling pathways.

OBJECTIVE

To assess effects of in vitro meloxicam exposure on metabolism in articular chondrocytes from dogs with naturally occurring osteoarthritis.

METHODS

Femoral head cartilage from 16 dogs undergoing total hip replacement.

METHODS

Articular cartilage samples were obtained. Tissue sulfated glycosaminoglycan (SGAG), collagen, and DNA concentrations were measured. Collagen, SGAG, chondroitin sulfate 8<em>4</em>6, NO, prostaglandin E2 (PGE2), and matrix metalloproteinase (MMP)-2, MMP-3, MMP-9, and MMP-13 concentrations in culture medium were analyzed. Aggrecan, collagen II, MMP-2, MMP-3, MMP-9, MMP-13, ADAM <em>metallopeptidase</em> with thrombospondin type 1 motif (ADAMTS)-<em>4</em>, ADAMTS-5, tissue <em>inhibitor</em> of metalloproteinase (<em>TIMP</em>)-1, <em>TIMP</em>-2, <em>TIMP</em>-3, interleukin-1β, tumor necrosis factor-α, cyclooxygenase-1, cyclooxygenase-2, and inducible nitric oxide synthase gene expression were evaluated. Comparisons between tissues cultured without (control) and with meloxicam at concentrations of 0.3, 3.0, and 30.0 μg/mL for up to 30 days were performed by means of repeated-measures analysis.

RESULTS

Meloxicam had no effect on chondrocyte SGAG, collagen, or DNA concentrations. Expression of ADAMTS-5 was significantly decreased in all groups on all days, compared with the day 0 value. On day 3, culture medium PGE2 concentrations were significantly lower in all meloxicam-treated groups, compared with values for controls, and values remained low. Culture medium MMP-3 concentrations were significantly lower on day 30 than on day 3 in all meloxicam-treated groups.

CONCLUSIONS

Results suggested that in vitro meloxicam treatment of osteoarthritic canine cartilage for up to 30 days did not induce matrix degradation or stimulate MMP production. Meloxicam lowered PGE2 release from this tissue, and effects on tissue chondrocyte content and matrix composition were neutral.