Downregulation of long non-coding RNA MEG3 in nasopharyngeal carcinoma.

In our previous whole-transcriptome sequencing analysis, downregulation of a long non-coding RNA, maternally expressed gene 3 (MEG3), was identified in NPC samples. This finding suggests the possible role of MEG3 as a tumor suppressor in this distinctive disease. In the present study, two MEG3 variants, AF119863 (MEG3-AF) and BX247998 (MEG3-BX), were found abundantly expressed in a normal nasopharyngeal epithelial cell line, NP69. Significant downregulation of MEG3-AF was further verified in a panel of NPC samples including xenografts and primary biopsies. MEG3 is an imprinted gene located within chromosome 14q32, a common deleted region in NPC. Both DNA copy number loss and aberrant promoter methylation contributed to MEG3 inactivation. Interestingly, MEG3 expression could successfully be rescued by the treatment of a demethylation agent. Besides, ectopic expression of MEG3 in NPC cell lines resulted in considerable repression of in vitro anchorage-independent growth and in vivo tumorigenicity, in addition to significant inhibition in cell proliferation, colony formation, and induction of cell cycle arrest. Finally, we revealed the association between MEG3 activity and the p53 signaling cascade. Our findings characterize MEG3 as a tumor suppressive long non-coding RNA in NPC and encourage the development of precise long non-coding RNA-targeted epigenetic therapy against this malignancy. © 2016 Wiley Periodicals, Inc.

Considerations for the Selection of Time-Limited Walk Tests Poststroke: A Systematic Review of Test Protocols and Measurement Properties.

BACKGROUND AND PURPOSE: Systematic reviews of research evidence describing the quality and methods for administering standardized outcome measures are essential to developing recommendations for their clinical application. The purpose of this systematic review was to synthesize the research literature describing test protocols and measurement properties of time-limited walk tests in people poststroke. METHODS: Following an electronic search of 7 bibliographic data-bases, 2 authors independently screened titles and abstracts. One author identified eligible articles, and performed quality appraisal and data extraction. RESULTS: Of 12 180 records identified, 43 articles were included. Among 5 walk tests described, the 6-minute walk test (6MWT) was most frequently evaluated (n = 36). Only 5 articles included participants in the acute phase (<1 month) poststroke. Within tests, protocols varied. Walkway length and walking aid, but not turning direction, influenced 6MWT performance. Intraclass correlation coefficients for reliability were 0.68 to 0.71 (12MWT) and 0.80 to 1.00 (2-, 3-, 5- and 6MWT). Minimal detectable change values at the 90% confidence level were 11.4 m (2MWT), 24.4 m (5MWT), and 27.7 to 52.1 m (6MWT; n = 6). Moderate-to-strong correlations (≥0.5) between 6MWT distance and balance, motor function, walking speed, mobility, and stair capacity were consistently observed (n = 33). Moderate-to-strong correlations between 5MWT performance and walking speed/independence (n = 1), and between 12MWT performance and balance, motor function, and walking speed (n = 1) were reported. DISCUSSION AND CONCLUSIONS: Strong evidence of the reliability and construct validity of using the 6MWT poststroke exists; studies in the acute phase are lacking. Because protocol variations influence performance, a standardized 6MWT protocol poststroke for use across the care continuum is needed.Video Abstract available for more insights from the authors (see Supplemental Digital Content 1, http://links.lww.com/JNPT/A150).

Speed and distance requirements for community ambulation: a systematic review.

OBJECTIVE: To provide an overview of the research literature on distance and speed requirements for adults to walk outside the home. DATA SOURCES: We conducted a systematic review and searched PubMed, MEDLINE (Ovid), EMBASE, CINAHL, Scopus, PEDro, and The Cochrane Library from 1948 to May 2012, and other sources. Search terms included communities, walk, ambulation, and neighborhood. STUDY SELECTION: Full-text peer-reviewed articles written in English, French, or Spanish reporting distance and/or speed requirements for individuals walking outside the home were considered eligible. Two authors independently screened titles and abstracts. One author reviewed full-text articles to determine inclusion. Of the 3191 titles and abstracts screened, 15 studies (.47%) were selected for detailed review. One author appraised methodological quality. Inadequate description of the reliability of the measurement methods and the population of the town/city assessed was noted. DATA EXTRACTION: One author extracted data from included studies. A second reviewer independently verified extracted data for accuracy. DATA SYNTHESIS: Seven studies examining 24 community sites and crosswalks in the United States, Australia, and Singapore were included. Three sites with the largest mean distance requirements for adults to walk were club warehouses (677m), superstores (183-607m), and hardware stores (566m). Three sites with the lowest mean distance requirements were walking at the front (16m) and back (19m) of the house, and at cemeteries (18m). The average speed required to cross the street in the time of a walk signal varied from .44 to 1.32m/s. CONCLUSIONS: Distance and speed requirements for adults to walk in the community environment vary widely. Findings are relevant to judging capacity for community ambulation to carry out essential activities of daily living, educating patients, and setting rehabilitation goals.

Retinal nerve fiber layer measurements in myopia: An optical coherence tomography study.

PURPOSE: To evaluate the relationship between retinal nerve fiber layer (RNFL) thickness measured by optical coherence tomography (OCT) and the axial length/refractive error of the eye. METHODS: A total of 115 eyes of 115 healthy subjects, comprising 75 eyes with high myopia (spherical equivalent [SE] < -6.0 D) and 40 eyes with low to moderate myopia (SE between -6.0 D and -0.5D), were analyzed in this cross-sectional study. Total average and mean clock hour RNFL thicknesses were measured by OCT and compared between the two myopia groups. Associations between RNFL measurements and axial length and spherical equivalent were evaluated by linear regression analysis. RESULTS: The RNFL measurements were significantly lower in the high myopia group compared with those of the low-to-moderate myopia group at 12, 1, and 7 o'clock (right eye orientation). Apart from the temporal clock hours, significant correlations were evident between RNFL measurements and the axial length and spherical equivalent. The average RNFL thickness decreased with increasing axial length (r = -0.314, P = 0.001) and negative refractive power (r = 0.291, P = 0.002). A significant proportion of myopic eyes were classified as outside normal limits, with reference to the normative database. The most frequently abnormal sector was at 2 o'clock, where 16.5% of myopic eyes were outside normal limits. CONCLUSIONS: RNFL measurements vary with the axial length/refractive error of the eye. Analysis of RNFL thickness in the evaluation of glaucoma should always be interpreted with reference to the refractive status. Although the normative database provided by OCT has been helpful in identifying ocular diseases involving the RNFL, it may not be reliable in the analysis of myopic eyes.

Reference values for standardized tests of walking speed and distance: a systematic review.

OBJECTIVE: To provide an overview of the reference values and methodology used to obtain them for time- and distance-limited walk tests. METHODS: We performed a systematic review and searched PubMed, MEDLINE (Ovid), EMBASE, CINAHL, Scopus, PEDro, and The Cochrane Library from 1946 to May 2013. Full-text peer-reviewed articles written in English, French or Spanish were considered eligible. Two authors independently screened titles and abstracts. One author determined eligibility of full-text articles, appraised methodological quality, and extracted data. A second author independently verified the accuracy of extracted data. RESULTS: Of the 41 eligible studies reviewed, 25 failed to describe the method used to select participants and 10 had an inadequate sample size. Twenty-five studies provided reference values for one time-limited walk test (6-min walk test (6 MWT)) and 18 studies provided reference values for 15 distance-limited walk tests. Across studies, walk test distances ranged from 3m to 40m. Descriptive values and reference equations for the 6 MWT were reported in 15 and 20 studies, respectively. Across 43 regression equations (median R(2)=0.46), age (98%) and sex (91%) were most frequently included. The equation yielding the maximum R(2) value (0.78) included age, height, weight and percentage of predicted maximum heart rate. Among six unique regression equations for distance-limited walk tests (median R(2)=0.17), sex (83%), age (67%) and weight (67%) were most frequently included. The equation yielding the maximum R(2) value (0.25) included age and sex. CONCLUSIONS: Reference values reported for these tests provide a basis for classifying walking capacity as within normal limits, determining the magnitude of deficit, educating clients, setting rehabilitation goals, and planning studies.

Analysis of pesticides and PCB congeners in serum by GC/MS with SPE sample cleanup.

This paper describes a simple procedure for the quantitative analysis of 25 polychlorinated biphenyl congeners (PCB) in serum. They are PCB 70, 74, 77, 87, 99, 101, 118, 126, 128, 138, 151, 153, 156, 169, 170, 180, 183, 187, 191, 194, 205, 206, 208 and 209. In addition, 15 pesticides are analyzed in the same extract: aldrin, p,p'-DDE, p,p'-DDT, dieldrin, endrin, endosulfan I, heptachlor, hexachlorobenzene, alpha-hexachlorocyclohexane, beta-hexachlorocyclohexane, lindane, methoxychlor, mirex, pentachlorobenzene, and 1,2,3,4-tetrachlorobenzene. The procedure involved sample cleanup using a combination of C18 and NH(2) solid phase extraction cartridges. The instrumental technique was gas chromatography/mass spectrometry with both electron ionization and negative chemical ionization in the selected ion monitoring mode. Concentration of sub-ng/ml level was attainable with 2 ml of serum sample.

Over-expression of miR-106b promotes cell migration and metastasis in hepatocellular carcinoma by activating epithelial-mesenchymal transition process.

Hepatocellular carcinoma (HCC) is one the the most fatal cancers worldwide. The poor prognosis of HCC is mainly due to the developement of distance metastasis. To investigate the mechanism of metastasis in HCC, an orthotopic HCC metastasis animal model was established. Two sets of primary liver tumor cell lines and corresponding lung metastasis cell lines were generated. In vitro functional analysis demonstrated that the metastatic cell line had higher invasion and migration ability when compared with the primary liver tumor cell line. These cell lines were subjected to microRNA (miRNAs) microarray analysis to identify differentially expressed miRNAs which were associated with the developement of metastasis in vivo. Fifteen human miRNAs, including miR-106b, were differentially expressed in 2 metastatic cell lines compared with the primary tumor cell lines. The clinical significance of miR-106b in 99 HCC clinical samples was studied. The results demonstrated that miR-106b was over-expressed in HCC tumor tissue compared with adjacent non-tumor tissue (p = 0.0005), and overexpression of miR-106b was signficantly correlated with higher tumor grade (p = 0.018). Further functional studies demonstrated that miR-106b could promote cell migration and stress fiber formation by over-expressing RhoGTPases, RhoA and RhoC. In vivo functional studies also showed that over-expression of miR-106b promoted HCC metastasis. These effects were related to the activation of the epithelial-mesenchymal transition (EMT) process. Our results suggested that miR-106b expression contributed to HCC metastasis by activating the EMT process promoting cell migration in vitro and metastasis in vivo.

14-3-3sigma expression as a prognostic marker in undifferentiated nasopharyngeal carcinoma.

In this study, we investigated the expression of 14-3-3sigma tumor suppressor gene in a panel of NPC cell lines, xenografts and primary tumors. Our objective was to determine the correlation between 14-3-3sigma expression and clinical outcome in NPC. We detected reduced 14-3-3sigma expression in 5/6 NPC tumor lines by quantitative RT-PCR and Western blotting. By immunohistochemical staining, significant down-regulation of 14-3-3sigma was also found in 26/72 (36.1%) primary tumors of NPC patients, who were treated with curative radiotherapy. Promoter methylation was confirmed in a subset of primary tumors by methylation-specific PCR analysis. Importantly, we demonstrated that 14-3-3sigma expression is significantly associated with both overall survival (OS) and cancer-specific survival (CSS), but not with the clinical staging of NPC patients. The low 14-3-3sigma expression was associated with improved overall (p=0.029) and cancer-specific survival (p=0.042) on univariate analysis. 14-3-3sigma expression and staging were also independent variables to all the prognostic factors under multivariate analysis. In conclusion, low expression of 14-3-3sigma appears to be a valuable marker for better survival in patient with NPC. These results provide the evidence that 14-3-3sigma expression is a significant prognostic factor for NPC patients.

Epigenetic inactivation of TSLC1 gene in nasopharyngeal carcinoma.

Deletion of 11q23 is a common genetic aberration in nasopharyngeal carcinoma (NPC). Multiple candidate tumor suppressor genes (TSG) were mapped to this region but few of them were investigated in NPC. TSLC1 (tumor suppressor in lung cancer) is recently reported to be a putative TSG on 11q23. This gene was found to be inactivated by promoter hypermethylation in non-small cell lung carcinoma (NSCLC), liver cancer, and breast cancer. To study the role of TSLC1 gene in NPC tumorigenesis, we screened for mutations and aberrant methylation of TSLC1 gene in 5 NPC cell lines, 3 NPC xenografts, and 38 primary NPC cases. No somatic mutations of TSLC1 were detected in the NPC samples, but a 9-bp (CCACCACCA) deletion in exon 8 was found in a primary NPC and its corresponding blood sample. Bisulfite sequencing revealed aberrant methylation of TSLC1 promoter in four NPC cell lines. Loss of TSLC1 gene expression was found in two cell lines (HK-1 and CNE-2) with dense methylation. Expression of this gene was restored in these cell lines after treatment with demethylating agent 5-aza-2'-deoxycytidine. Our results showed that silencing of TSLC1 gene expression in NPC was associated with promoter hypermethylation. Promoter hypermethylation of TSLC1 gene was further illustrated in 34.2% (13/38) of primary NPCs. No aberrant promoter methylation was found in any of the four investigated normal nasopharyngeal epithelia. Frequent epigenetic inactivation of TSLC1 gene in NPC suggested that this gene is one of the target tumor suppressor genes of this endemic cancer.