VNN3 is a potential novel biomarker for predicting prognosis in clear cell renal cell carcinoma

Although pathological observations provide approximate prognoses, it is difficult to achieve prognosis in patients with existing prognostic factors. Therefore, it is very important to find appropriate biomarkers to achieve accurate cancer prognosis. Renal cell carcinoma (RCC) has several subtypes, the discrimination of which is crucial for proper treatment. Here, we present a novel biomarker, VNN3, which is used to prognose clear cell renal cell carcinoma (ccRCC), the most common and aggressive subtype of kidney cancer. Patient information analyzed in our study was extracted from The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) cohorts. VNN3 expression was considerably higher in stages III and IV than in stages I and II. Moreover, Kaplan–Meier curves associated high VNN3 expression with poor prognoses (TCGA, p?p?=?.00076), confirming that ccRCC prognosis can be predicted via VNN3 expression patterns. Consistent with all patient results, the prognosis of patients with higher VNN3 expression was worse in both low stage (I and II) and high stage (III and IV) (TCGA, p < 0.0001 in stage I and II; ICGC, p = 0.028 in stage I and II; TCGA, p = 0.005 in stage III and IV). Area under the curve and receiver operating characteristic curves supported our results that highlighted VNN3 expression as a suitable ccRCC biomarker. Multivariate analysis also verified the prognostic performance of VNN3 expression (TCGA, p?p?=?.017). Altogether, we suggest that VNN3 is applicable as a new biomarker to establish prognosis in patients with ccRCC.     

Light Exposure Among Adolescents With Delayed Sleep Phase Disorder: A Prospective Cohort Study

The objective of this study was to compare light exposure and sleep parameters between adolescents with delayed sleep phase disorder (DSPD; n?=?16, 15.3?±?1.8 yrs) and unaffected controls (n?=?22, 13.7?±?2.4 yrs) using a prospective cohort design. Participants wore wrist actigraphs with photosensors for 14 days. Mean hourly lux levels from 20:00 to 05:00?h and 05:00 to 14:00?h were examined, in addition to the 9-h intervals prior to sleep onset and after sleep offset. Sleep parameters were compared separately, and were also included as covariates within models that analyzed associations with specified light intervals. Additional covariates included group and school night status. Adolescent delayed sleep phase subjects received more evening (p?<?.02, 22:00–02:00?h) and less morning (p?<?.05, 08:00–09:00?h and 10:00–12:00?h) light than controls, but had less pre-sleep exposure with adjustments for the time of sleep onset (p?<?.03, 5–7?h prior to onset hour). No differences were identified with respect to the sleep offset interval. Increased total sleep time and later sleep offset times were associated with decreased evening (p?<?.001 and p?=?.02, respectively) and morning (p?=?.01 and p?<?.001, respectively) light exposure, and later sleep onset times were associated with increased evening exposure (p?<?.001). Increased total sleep time also correlated with increased exposure during the 9?h before sleep onset (p?=?.01), and a later sleep onset time corresponded with decreased light exposure during the same interval (p?<?.001). Outcomes persisted regardless of school night status. In conclusion, light exposure interpretation requires adjustments for sleep timing among adolescents with DSPD. Pre- and post-sleep light exposures do not appear to contribute directly to phase delays. Sensitivity to morning light may be reduced among adolescents with DSPD. (Author correspondence: auger.raymond1@mayo.edu)     

Forward head posture is associated with pressure pain threshold and neck pain duration in university students with subclinical neck pain

Abstract

Objective: The aims of this study are to investigate the association between: (i) forward head posture (FHP) and pressure pain thresholds (PPTs); (ii) FHP and maladaptive cognitive processes; and (iii) FHP and neck pain characteristics in university students with subclinical neck pain.

Materials/methods: A total of 140 university students, 90 asymptomatic and 50 with subclinical neck pain, entered the study. Demographic data, anthropometric data, FHP, and PPTs were collected for both groups. In addition, pain characteristics, pain catastrophizing, and fear of movement were assessed for participants with neck pain. FHP was characterized by the angle between C7, the tragus of the ear, and the horizontal line. Correlation analysis and multivariate regression analysis were conducted.

Results: Participants with subclinical neck pain showed significantly lower PPTs than participants without neck pain (p?<?.05), but similar FHP (p?>?.05). No significant association was found between FHP and PPTs in the asymptomatic group. In the group of participants with subclinical neck pain, PPTs at the right trapezius and neck pain duration explained 19% of the variance of FHP (R²?=?0.23; adjusted R²?=?0.19; p?<?.05).

Conclusion: This study suggests that FHP is not associated with PPTs in asymptomatic university students. In university students with subclinical neck pain, increased FHP was associated with right trapezius hypoalgesia and with neck pain of shorter duration. These findings are in contrast with current assumptions on the association between neck pain and FHP.     

Curcumin and its derivatives inhibit 2,3,7,8,-tetrachloro-dibenzo-p-dioxin-induced expression of drug metabolizing enzymes through aryl hydrocarbon receptor-mediated pathway

Abstract

Certain dioxins, including 2,3,7,8,-tetrachloro-dibenzo-p-dioxin (TCDD), are exogenous ligands for an aryl hydrocarbon receptor (AhR) and induces various drug-metabolizing enzymes. In this study, we examined the effect of curcumin on expression of drug-metabolizing enzymes through the AhR and NF-E2 related factor 2 (Nrf2) pathways. Curcumin dose-dependently inhibited TCDD-induced expression of phase I enzyme cytochrome P450 1A1 (CYP1A1) and phase II enzymes NAD(P)H:quinone oxidoreductase-1 (NQO1) and heme oxygenase 1 (HO-1) but not tert-butyl hydroquinone-induced NQO1 and HO-1, suggesting that curcumin inhibited only AhR pathway, but not Nrf2 one directly. Furthermore, we used 14 curcumin derivatives and obtained the correlation between hydrophobicity of the compounds and suppressive effect against AhR transformation. Results from the quantitative structure active correlative analysis indicated that methoxy groups and β-diketone structure possessing keto-enol tautomerism in curcumin were necessary to inhibit AhR transformation, and the addition of methyl and methoxy group(s) to the curcumin increased the inhibition effect.     

SIRT1 and circadian gene expression in pancreatic ductal adenocarcinoma: Effect of starvation

Pancreatic cancer (PC), the fourth leading cause of cancer-related deaths, is characterized by high aggressiveness and resistance to chemotherapy. Pancreatic carcinogenesis is kept going by derangement of essential cell processes, such as proliferation, apoptosis, metabolism and autophagy, characterized by rhythmic variations with 24-h periodicity driven by the biological clock. We assessed the expression of the circadian genes ARNLT, ARNLT2, CLOCK, PER1, PER2, PER3, CRY1, CRY2 and the starvation-activated histone/protein deacetylase SIRT1 in 34 matched tumor and non-tumor tissue specimens of PC patients, and evaluated in PC derived cell lines if the modulation of SIRT1 expression through starvation could influence the temporal pattern of expression of the circadian genes. We found a significant down-regulation of ARNLT (p?=?0.015), CRY1 (p?=?0.013), CRY2 (p?=?0.001), PER1 (p?<?0.0001), PER2 (p?<?0.001), PER3 (p?=?0.001) and SIRT1 (p?=?0.017) in PC specimens. PER3 and CRY2 expression levels were lower in patients with jaundice at diagnosis (?<?0.05). Having adjusted for age, adjuvant therapy and tumor stage, we evidenced that patients with higher PER2 and lower SIRT1 expression levels showed lower mortality (p?=?0.028). Levels and temporal patterns of expression of many circadian genes and SIRT1 significantly changed upon serum starvation in vitro, with differences among four different PC cell lines examined (BXPC3, CFPAC, MIA-PaCa-2 and PANC-1). Serum deprivation induced changes of the overall mean level of the wave and amplitude, lengthened or shortened the cycle time and phase-advanced or phase-delayed the rhythmic oscillation depending on the gene and the PC cell line examined. In conclusion, a severe deregulation of expression of SIRT1 and circadian genes was evidenced in the cancer specimens of PC patients, and starvation influenced gene expression in PC cell lines, suggesting that the altered interplay between SIRT1 and the core circadian proteins could represent a crucial player in the process of pancreatic carcinogenesis.     

Research on association between variants and haplotypes of TRPV1 and TRPA1 genes with growth traits in three cattle breeds

The objective of this study was to examine the association of transient receptor potential vanilloid 1 (TRPV1) and transient receptor potential ankyrin 1 (TRPA1) genes polymorphisms with growth traits in three Chinese cattle breeds (Jiaxian red cattle, Qinchuan cattle and Luxi cattle). Through experiments we identified three single nucleotide polymorphisms (SNPs) in these three cattle breeds TRPV1 and TRPA1 genes using PCR-SSCP, (forced) PCR-RFLP methods. Three of these polymorphisms are all synonymous mutation which includes (NW_003104493.1: 30327 C?>?T), (NW_003104493.1: 33394 A?>?G) and (NW_003104493.1: 38471?G?>?A) are in exons. The other three polymorphisms are located at 3'UTR. Furthermore, we evaluated the haplotype frequency and the statistical analyses indicated that these SNPs of TRPV1 and TRPA1 genes were associated with bovine body height, body length, waist angle width, hucklebone width, cross ministry height, chest qingwidth (p?<?0.05) and recommendation height, cannon circumference (p?<?0.01) of Qingchuan cattle; body length, waist angle width (p?<?0.05) of Jiaxian red cattle; body weight, Body length, cannon circumference, chest circumference (p?<?0.05) and body height (p?<?0.01) of Luxi cattle. Our result confirms the polymorphisms in the TRPV1 and TRPA1 genes are associated with growth traits that may be used for marker-assisted selection (MAS) in three beef breeds programs.     

The severity of chewing disorders is related to gross motor function and trunk control in children with cerebral palsy

Abstract

Purpose: The frequency of chewing disorders increases with decreasing level of gross motor function in children with cerebral palsy (CP). Besides its frequency, the severity of chewing disorders is also important. The aim of this study was to determine the relationship between chewing performance level and gross motor function, and trunk postural control in children with CP.

Materials and methods: The study included 119 children with CP (age 2–10 years). Chewing performance level was determined by the Karaduman Chewing Performance Scale (KCPS). The Gross Motor Function Classification System (GMFCS) was used to determine the level of gross motor function. Segmental Assessment of Trunk Control (SATCo) was used to measure trunk control.

Results: Children with spastic CP with a median age of 4?years were evaluated, of which 50.4% were male. The percentages of patients classified to GMFCS levels I to V were 43.7%, 6.7%, 9.2%, 5.0%, and 35.3%, respectively. The median KCPS score was 3 (min?=?0, max?=?4). A good correlation was found between KCPS and GMFCS (p?<?.001, r?=?0.70). Negative, excellent correlations between KCPS and SATCo static, SATCo active, and SATCo reactive postural controls were found (p?<?.001, r?=?–0.75, r?=?–0.77, r?=?–0.79; respectively).

Conclusions: The severity of chewing disorders is related to the level of gross motor function and trunk postural control in children with CP.

Clinical trial number: NCT03241160     

应用慢病毒shRNA文库结合二代测序筛选白血病细胞系增殖相关lncRNA北大核心CSCD

长链非编码RNA(long non-coding RNA,lncRNA)是包括细胞增殖在内的许多细胞过程的重要调节因子。虽然已有研究表明多种lncRNA在造血系统恶性肿瘤的发生发展过程中发挥重要作用,但是缺少一个更全面和无偏倚的方法同时研究多个lncRNA中对白血病细胞系产生功能性影响的lncRNA。在此,我们利用短发夹RNA(short hairpin RNA,shRNA)文库结合高通量测序的方法,筛选对白血病细胞系增殖有影响的lncRNA,确定了74个候选lncRNAs。从中选取lncRNA C20orf204-203作为验证研究对象,发现C20orf204-203在K562和THP-1细胞系中均定位于胞质,敲降C20orf204-203的K562和THP-1细胞系增殖能力降低,早期凋亡细胞增加,BAD基因在mRNA水平上表达量增加,TP53、BCL2蛋白表达量下降,在THP-1细胞系中Caspase 3蛋白表达量减少,激活型Caspase 3蛋白表达量上升,但是二者变化在两种细胞系中不一致。结果表明,在白血病细胞系中敲降lncRNA C20orf204-203会使细胞增殖能力降低。但其在不同细胞系作用途径和机制可能存在差异。这一研究表明了利用shRNA文库结合高通量测序大规模研究lncRNA在白血病细胞系中发挥作用的可行性。     

Comparison of unpleasant and pain thresholds of thermal stimuli in the orofacial regions: a psychophysical study using quantitative sensory testing in healthy young men

Abstract

Purpose/Aim: To gain a better understanding of the psychophysics of thermal pain perception in a clinical setting, this study investigated whether thermal thresholds of unpleasantness are different from pain thresholds of cold and heat stimuli. Of particular interest was the relationship between unpleasantness and pain thresholds for cold vs heat stimuli.

Material and methods: Thirty healthy male volunteers (mean age 26.1?years, range 23 to 32?years) participated. Thermal detection, cold pain (CPT) and heat pain (HPT) thresholds were measured at 5 trigeminal sites by the method of limits using quantitative sensory testing (QST), followed by cold unpleasant (CUT) and heat unpleasant (HUT) thresholds.

Results: The temperatures at which individuals first reported thermal sensations as unpleasant or painful substantially differed among subjects. CUT exhibited a higher mean value with less variability than CPT, and HUT presented a lower mean than HPT (p?<?.001). As with CPT, CUT did not show any significant difference between the test sites. On the other hand, HUT, like HPT, exhibited site differences (p?<?.001). There was moderate correlation between CUT and CPT, whereas HUT and HPT were strongly correlated. The relationship between unpleasant and pain thresholds of cold vs heat stimuli was significantly different even when controlling for test site variability (p?<?.001).

Conclusion: These findings indicate that unpleasant and pain thresholds to thermal stimuli differ in healthy young men. Of particular note is the distinct relationship of unpleasant and pain thresholds of cold vs heat stimuli, revealing the thermal difference in temperature transition from unpleasantness to pain.     

Downregulation of the circadian Period family genes is positively correlated with poor head and neck squamous cell carcinoma prognosis

ABSTRACT

We investigated the relationship between head and neck squamous cell carcinoma (HNSCC) and the mRNA and protein expression levels of the circadian genes of the Period (Per) family, Per1, Per2 and Per3. Tissue sections of HNSCC and normal head and neck tissues from two patient cohorts from two different hospitals were collected to assess the mRNA and protein expressions of the three Per family genes using real-time quantitative PCR (RT-PCR) and immunohistochemistry (IHC). The clinicopathological features and disease prognosis for the latter cohort were analyzed through IHC and statistical methods. Protein positive expression levels of the three Per family genes in HNSCC tissues was found to be approximately two times lower than that in normal tissues (p < .01). Moreover, patients with locally advanced HNSCC showed significantly greater downregulation of Per1, Per2 and Per3 mRNA expression levels as compared to patients with early-stage cancer (p < .05). Immunohistochemical examination of HNSCC patient tissues revealed a positive correlation between the Per family protein expression and the clinical tumor staging (p < .05). In addition, the Per protein-positive expression group showed higher 3-year survival rates [overall survival (OS) and progression-free survival (PFS)] as assessed by Kaplan-Meier plots and statistical analysis (p < .05). Our findings confirm the positive correlation between Per family gene expression and survival outcomes and support their role as prognostic markers for HNSCC.     

Alternation of extracellular matrix remodeling and apoptosis by activation of the aryl hydrocarbon receptor pathway in human periodontal ligament cells

It is well known that the aryl hydrocarbon receptor (AhR) is involved in the toxicity of halogenated aromatic hydrocarbons (HAH) and polycyclic aromatic hydrocarbons (PAH). Recent experiments have shown the induction of impaired tooth and hard‐tissue formation by AhR pathway activation, however, the effect on periodontal ligament (PDL) tissue remains unclear. Here, we investigated the effects of benzo(a)pyrene (BaP), a member of PAH, on the extracellular matrix (ECM) remodeling‐related molecules, collagen type I (COL‐I), matrix metalloproteinase‐1 (MMP‐1), alpha‐smooth muscle actin (α‐SMA) expression, and apoptosis in two different human periodontal ligament cells (HPDLCs). The transduction of AhR from the cytoplasm to the nucleus and the increase of AhR‐responsive genes; that is, cytochrome P450 1A1 (CYP1A1), cytochrome P450 1B1 (CYP1B1), and aryl‐hydrocarbon receptor repressor (AhRR), expression was induced by BaP exposure in both HPDLCs. BaP treatment significantly enhanced MMP‐1 mRNA expression and MMP‐1 protein production, while markedly suppressing COL‐I and a‐SMA mRNA expression in both HPDLCs. Furthermore, these BaP‐treated HPDLCs fell into apoptotic cell death as evidenced by induction in annexin V and caspase‐3/7 staining and reduction of total cell number and Bcl‐2 mRNA expression. Thus, BaP exposure altered the expression of ECM‐related molecules and induced apoptosis in HPDLCs through activation of the AhR pathway. Overactivity of the AhR pathway may induce an inappropriate turnover of PDL tissue via disordered ECM remodeling and apoptosis in PDL cells. J. Cell. Biochem. 113: 3093–3103, 2012. © 2012 Wiley Periodicals, Inc.     

JTC-801 exerts anti-proliferative effects in human osteosarcoma cells by inducing apoptosis

Background: The research of G protein-coupled receptors (GPCRs) is a promising strategy for drug discovery. In cancer therapy, there is a need to discover novel agents that can inhibit proliferation and induce apoptosis in cancer cells. JTC-801 is a novel GPCR antagonist with the function of reversing pain and anxiety symptoms. This study aims to investigate the antitumor effects of JTC-801 on human osteosarcoma cells (U2OS) and elucidate the underlying mechanism.

Materials and methods: The Cell Counting Kit-8 assay was used to detect the viability of U2OS cells treated with JTC-801 in vitro. The cell apoptosis was evaluated using a flow cytometry assay with Annexin V-FITC/PI double staining. The inhibitory effect of JTC-801 on invasion and migration of U2OS cells were determined by the Transwell assays. Western blot assay was performed to measure the levels of proteins related to cell apoptosis and its mechanism.

Results: The JTC-801 significantly decreased the viability of U2OS cells (p?p?p?Conclusions: JTC-801 may exert osteosarcoma cell growth inhibition by promoting cell apoptosis, through PI3K/AKT signaling pathway participation.     

Thioredoxin-1 is required for the cardioprotecive effect of sildenafil against ischaemia/reperfusion injury and mitochondrial dysfunction in mice

Abstract

Sildenafil is a phosphodiesterase type 5 inhibitor which confers cardioprotection against myocardial ischaemia/reperfusion (I/R) injury. The aim of this study was to determine if Trx1 participates in cardioprotection exerted by sildenafil in an acute model of I/R, and to evaluate mitochondrial bioenergetics and cellular redox status. Langendorff-perfused hearts from wild type (WT) mice and a dominant negative (DN-Trx1) mutant of Trx1 were assigned to placebo or sildenafil (0.7?mg/kg i.p.) and subjected to 30?min of ischaemia followed by 120?min of reperfusion. WT?+?S showed a significant reduction of infarct size (51.2?±?3.0% vs. 30?±?3.0%, p < .001), an effect not observed in DN-Trx. After I/R, sildenafil preserved state 3 oxygen consumption from WT, but had a milder effect in DN-Trx1 only partially protecting state 3 values. Treatment restored respiratory control (RC) after I/R, which resulted 8% (WT) and 24% (DN-Trx1) lower than in basal conditions. After I/R, a significant increase in H₂O₂ production was observed both for WT and DN-Trx (WT: 1.17?±?0.13?nmol/mg protein and DN-Trx: 1.38?±?0.12?nmol/min mg protein). With sildenafil, values were 21% lower only in WT I/R. Treatment decreased GSSG levels both in WT and DN-Trx1. In addition, GSSG/GSH² ratio was partially restored by sildenafil. Also, an increase in p-eNOS/eNOS even before the myocardial ischaemia was observed with sildenafil, both in WT (14%, p > .05) and in DN-Trx (35%, p < .05). Active Trx1 is required for the onset of the cardioprotective effects of sildenafil on I/R injury, together with the preservation of cellular redox balance and mitochondrial function.     

Design,synthesis and biological evaluation of 4-bromo-N-(3,5-dimethoxyphenyl)benzamide derivatives as novel FGFR1 inhibitors for treatment of non-small cell lung cancer

A series of 4-bromo-N-(3,5-dimethoxyphenyl)benzamide derivatives were designed and synthesised as novel fibroblast growth factor receptor-1 (FGFR1) inhibitors. We found that one of the most promising compounds, C9, inhibited five non-small cell lung cancer (NSCLC) cell lines with FGFR1 amplification, including NCI-H520, NCI-H1581, NCI-H226, NCI-H460 and NCI-H1703. Moreover, the IC₅₀ values for the compound C9 were 1.36?±?0.27?µM, 1.25?±?0. 23?µM, 2.31?±?0.41?µM, 2.14?±?0.36?µM and 1.85?±?0.32?µM, respectively. The compound C9 arrested the cell cycle at the G2 phase in NSCLC cell lines. The compound C9 also induced cellular apoptosis and inhibited the phosphorylation of FGFR1, PLCγ1 and ERK in a dose-dependent manner. In addition, molecular docking experiments showed that compound C9 binds to FGFR1 to form six hydrogen bonds. Taken together, our data suggested that the compound C9 represented a promising lead compound-targeting FGFR1.