Serum biomarkers have not been fully incorporated into clinical use for the diagnosis of renal cell carcinoma (RCC). The recent discovery of long noncoding RNAs (lncRNAs), which have been reported in a variety of cancer types, suggested a promising new class of biomarkers for tumour diagnosis. The aim of our study was to evaluate whether the levels of circulating lncRNAs could be used as a tumour marker to discriminate between clear cell RCC (ccRCC) patients and healthy controls. Serum samples were collected from 71 ccRCC patients including 62 age- and sex-matched healthy controls and 8 patients with benign renal tumours. Eighty-two cancer-associated lncRNAs were assessed by reverse transcription and quantitative polymerase chain reaction in paired tissues and serum. A 5-lncRNA signature, including lncRNA-LET, PVT1, PANDAR, PTENP1 and linc00963, were identified and validated in the training set and testing set, respectively. The receiver operating characteristic curves for this serum 5-lncRNA signature were 0.900 and 0.823 for the two sets of serum samples. Moreover, five-minus-one lncRNA signatures demonstrated that none of the lncRNAs had a higher area under the curve than the others in either set. A risk model for the serum 5-lncRNA signature also determined that benign renal tumours can be distinguished from ccRCC samples. This work may facilitate the detection of ccRCC and serve as the basis for further studies of the clinical value of serum lncRNAs in maintaining surveillance and forecasting prognosis.
Polyploidization is an evolutionarily rare but important mechanism in both plants and animals because it increases genetic diversity. Goldfish of the Carassius auratus species complex can be tetraploids, hexaploids and octaploids. Polyploidization events have occurred repeatedly in goldfish, yet the extent of this phenomenon and its phyletic history are poorly understood. We explore the origin, tempo and frequency of polyploidization in Chinese and Japanese goldfish using both mitochondrial (mtDNA) and nuclear DNA sequences from up to 1202 individuals including the outgroup taxon, Cyprinus carpio. Analyses of de novo nuclear gene data resolve two clusters of alleles and the pattern supports the prior hypothesis of an ancient allotetraploidization for Carassius. Alleles shared by tetraploid and hexaploid individuals indicate recent autoploidizations within the C. auratus complex. Sympatric tetraploids and hexaploids share mtDNA haplotypes and these frequently occur independently within six well-supported lineages and sublineages on a small spatial scale. Gene flow estimates (F st values) indicate that hexaploids differ only slightly from sympatric tetraploids, if at all. In contrast, allopatric populations of tetraploids and hexaploids differ from one another to a far greater extent. Gene flow between sampled localities appears to be limited. Coalescence-based time estimations for hexaploids reveal that the oldest lineage within any sampled locality is around one million years old, which is very young. Sympatric, recurrent autoploidization occurs in all sampled populations of the C. auratus complex. Goldfish experience polyploidization events more frequently than any other vertebrate.
Fermented cottonseed meal (FCSM) is widely used in poultry diets in China. This study was conducted to investigate the effect of FCSM on lipid-related gene expression in broilers. Initially, 180 broiler chickens (21-days-old, equal number of males and females) were randomly divided into three groups, with six pens per group and 10 birds per pen. The chickens in the control group were fed a diet containing unfermented cottonseed meal, and those in the treatment groups were fed with diets including either CSM fermented by Candida tropicalis (Ct group) or CSM fermented by Candida tropicalis plus Saccharomyces cerevisae (Ct-Sc group) until 64 days old. The results revealed that, compared with the control group (p<0.05), the mRNA expression of peroxisome proliferator-activated receptor alpha (PPAR-α) and lipoprotein lipase (LPL) were upregulated in the livers of Ct-Sc males. The expression of PPAR-α was also upregulated in the livers of Ct females. The expression levels of acetyl CoA carboxylase (ACC) and LPL in the liver of males and the expression of PPAR-α in the liver of females were significantly different between the Ct and Ct-Sc groups (p<0.05). However, gene expressions of fatty acid synthase (FAS) and liver fatty acid-binding protein (L-FABP) in the liver were not altered when the broilers were fed FCSM-supplemented diets (p>0.05). Likewise, the expressions of peroxisome proliferator-activated receptor gamma (PPAR-γ) and LPL in the abdominal fat were not altered by the FCSMsupplemented diets (p>0.05). The results in this study indicate that CSM fermented by Candida tropicalis and Saccharomyces cerevisiae effectively regulated the genes involved in fatty acid β-oxidation and triglyceride hydrolysis in male broiler chickens. Furthermore, the effects of the FCSM-supplemented diets were significantly different between bird sexes and between yeast strains used in the fermentation process.
Metabolic stress is a common phenomenon in solid tumors. Compensatory mechanisms to overcome metabolic stress (glucose deprivation) are vital to tumor cell survival. The histone demethylase Jumonji domain-containing protein 2B (JMJD2B) is vital for the growth and progression of various cancers. However, the role of JMJD2B during glucose deprivation remains unclear. Our aim was to examine the function of JMJD2B in glucose-deprived colon cancer cells and the involvement of extracellular signal-regulated kinase (ERK) and glucose transporter 1 (GLUT1). Our study demonstrated that JMJD2B expression was upregulated via ERK phosphorylation during glucose deprivation. Further, the cell viability assay showed that the effect of p-ERK on the viability of colon cancer cells was at least partly dependent on JMJD2B expression. Glucose deprivation increased interaction between JMJD2B and p-ERK, as demonstrated by the co-immunoprecipitation and fluorescence assays. Glucose deprivation also resulted in phosphorylation of JMJD2B by p-ERK, as shown by the immunoprecipitation assay and western blotting analysis. In addition, the phosphorylation of JMJD2B via p-ERK at Thr305, Ser352, Ser566 and Thr1065 contribute to JMJD2B stability. p-ERK stabilizes the JMJD2B protein level by protecting JMJD2B from ubiquitination and proteasome degradation. We found that knockdown of JMJD2B significantly impaired colon cancer cell viability, which is accompanied by a significant reduction in glucose uptake and lactate production. Furthermore, knockdown of JMJD2B after glucose deprivation caused decreased level of GLUT1 through increasing H3K9 tri-methylation levels on its promoter, demonstrated by chromatin immunoprecipitation assay. Moreover, targeting JMJD2B in xenograft tumors also decreased the GLUT1 level. Finally, a positive correlation was observed between p-ERK, JMJD2B and GLUT1 expression in 85 human colon cancer tissue specimens. These results indicated that p-ERK-mediated phosphorylation and stabilization of JMJD2B during glucose deprivation contributes to its role in glucose uptake and cell viability, which may be modulated through epigenetically upregulation of GLUT1.
Multi-bolt joint distributed around the connecting members are generally adopted to meet the high-performance assembly requirements in aerospace, energy and power industries. However, the initial preload could be low due to non-optimized preload sequence and bolt stress relaxation, especially at elevated temperature. Thus, it is necessary to take elastic interaction and bolt stress relaxation into account before jointing. In this article, a general multi-bolt elastic interaction with bolt stress relaxation is modelled analytically. First, the multi-bolt joint is characterized by ‘spring-node’ model and elastic interaction stiffness. Second, the bolt residual preload can be estimated according to linear superposition of elastic interaction and bolt stress relaxation under the condition of node displacement equilibrium. Further, the influence of preloading sequence and bolt stress relaxation on residual preload distribution was numerically analyzed using a typical circular ring with 8-bolt joint. Two bolts’ preloading sequences were planned, star sequence and counterclockwise sequence, respectively. The bolt creep simulation time was set as 10 h using the power-model at intermediate temperature. From comparison, the predicted results using the developed model were consistent with the FE simulations both with and without bolt stress relaxation.
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Identification of genes that are specifically expressed in the adult testis or the fetal testis is important for the study of genes related to the development of the testis. In this study, a human testis cDNA microarray was established. PCR products of 9216 clones from a human testis cDNA library were dotted on a nylon membrane; mRNA from adult and fetal testes were purified and probes were prepared by a reverse transcription reaction with testis mRNA as template. The microarray was hybridized with probes of adult and fetal testes, and 96.8 and 95.4% of clones were positive, respectively. In total, 731 clones were differentially expressed: 592 were highly expressed in adult testis and 139 were highly expressed in fetal testis. Among these genes, a new reticulon (Rtn)-like gene was detected and named Rtn-T. Rtn-T was highly expressed in adult human testis. The cDNA of Rtn-T contains 3491 bp and the putative protein had 968 amino acids. This protein is homologous to the six known members of the Rtn family (KIAA0886, Rtn xL, reticulon 4a, Nogo-A, Nogo-A short form, and brain my043) but was different at the 5' end. All homologues originate from one gene, and result from both different promotor regions and different splicing. Rtn-T lacks the first exon and contains a second exon that is lacking in the other homologues. Rtn-T is shorter than KIAA0886, Rtn xL, reticulon 4a and Nogo-A, but longer than the Nogo-A short form and brain my043. Sequence analysis showed that Rtn-T protein has two hydrophobic regions that may be membrane-spanning domains. Expression profiles showed that Rtn-T is specifically and strongly expressed in testis. The results of the present study indicate that the Rtn-T gene is differentially expressed in adult and fetal testes and encodes a membrane protein that may have a function in testis development.
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