Identification of polymorphisms associated with economic traits is important for successful marker-assisted selection in cattle breeding. The family of mammalian sirtuin regulates many biological functions, such as life span extension and energy metabolism. SIRT2, a most abundant sirtuin in adipocytes, acts as a crucial regulator of adipogenic differentiation and plays a key role in controlling adipose tissue function and mass. Here we investigated single nucleotide polymorphisms (SNPs) of bovine SIRT2 in 1226 cattle from five breeds and further evaluated the effects of identified SNPs on economically important traits of Nanyang cattle. Our results revealed four novel SNPs in bovine SIRT2, one was located in intronic region and the other three were synonymous mutations. Linkage disequilibrium and haplotype analyses based on the identified SNPs showed obvious difference between crossbred breed and the other four beef breeds. Association analyses demonstrated that SNPs g.17333C > T and g.17578A > G have a significantly effect on 18-months-old body weight of Nanyang population. Animals with combined genotype TTGG at the above two loci exhibited especially higher body weight. Our data for the first time demonstrated that polymorphisms in bovine SIRT2 are associated with economic traits of Nanyang cattle, which will be helpful for future cattle selection practices.
Japanese larch (Larix kaempferi) has been introduced in China at the end of the 19th century, and as one successful exotic species, is becoming the preferred coniferous in northern China and sub-tropical alpine region. The rotation age is about 25-28 years for L. kaempferi as pulpwood in Henan province. Waiting for even one-half rotation age for final evaluation will be inefficient due to accumulated testing costs and delayed return on investment, which suggests that selection at an early age is highly desirable for L. kaempferi improvement programs in Henan province. In this study, we determined age trends of genetic parameters and evaluated early selection efficiency for L. kaempferi in Henan province to find out the appropriate trait for early selection and its selection age.
To observe form and function changes of vascular endothelial cells (VEC) which were transplanted to the posterior surface of rhesus monkey cornea without Descemet's Membrane by anterior chamber injection, explore the feasibility of transplanting VEC to treat corneal endothelial injury, and find new method of corneal endothelial cell transplantation.
A fingerprint analysis method has been developed for characterization and discrimination of polysaccharides from different Ganoderma by high performance liquid chromatography (HPLC) coupled with chemometrics means. The polysaccharides were extracted under ultrasonic-assisted condition, and then partly hydrolyzed with trifluoroacetic acid. Monosaccharides and oligosaccharides in the hydrolyzates were subjected to pre-column derivatization with 1-phenyl-3-methyl-5-pyrazolone and HPLC analysis, which will generate unique fingerprint information related to chemical composition and structure of polysaccharides. The peak data were imported to professional software in order to obtain standard fingerprint profiles and evaluate similarity of different samples. Meanwhile, the data were further processed by hierarchical cluster analysis and principal component analysis. Polysaccharides from different parts or species of Ganoderma or polysaccharides from the same parts of Ganoderma but from different geographical regions or different strains could be differentiated clearly. This fingerprint analysis method can be applied to identification and quality control of different Ganoderma and their products.
This meta-analysis was designed to assess the overall performance of GnRHa in preserving the ovarian function in young women undergoing chemotherapy. Electronic literature databases including Pubmed, MEDLINE, Cochrane library, Embase, CNKI and Wanfang were searched for articles published till November, 2013. The articles written in both Chinese and English were considered. Only randomized controlled trials (RCTs) were selected. Main Outcome Measure was evaluated by assessing the post?chemotherapy ovarian function. A random-effects model was used to calculate the risk ratio (RR) and associated 95% confidence intervals (95% CI). Out of the eight RCTs including 621 patients, 321 women were treated with GnRHa during chemotherapy, 9.66% of whom suffered premature ovarian failure (POF). On the other hand, 26.67% of the remaining 300 women suffered POF. More women treated without GnRHa experienced post-chemotherapy POF, yielding an RR of 0.45 [chemotherapy plus GnRHa vs. chemotherapy alone, 95% confidence interval (CI) (0.22, 0.92)]. Based on the available studies, GnRHa plays an important role in the prevention of post-chemotherapy POF, but does not exhibit its protective effects in fertility.
Maxillary anterior segmental distraction osteogenesis (MASDO) is a recently used method for correction of severe maxillary retrusion in cleft lip and palate (CLP) patients. In this article, we evaluated the feasibility of MASDO using rigid external distraction (RED) and rapid orthodontic tooth movement to correct severe maxillary retrusion in CLP patients.
As the master regulator of adipogenesis, peroxisome proliferator-activated receptor gamma (PPARG) is required for the accumulation of adipose tissue and hence contributes to obesity. A previous study showed that the substitution of +20A>G in PPARG changed the 7(th) amino acid from Asp to Gly, creating a mutant referred to as PPARG Asp7Gly. In this study, association analysis indicated that PPARG Asp7Gly was associated with lower body height, body weight and heart girth in cattle (P<0.05). Overexpression of PPARG in NIH3T3-L1 cells showed that the Asp7Gly substitution may cause a decrease in its adipogenic ability and the mRNA levels of CIDEC (cell death-inducing DFFA-like effector c) and aP2, which are all transcriptionally activated by PPARG during adipocyte differentiation. A dual-luciferase reporter assay was used to analyze the promoter activity of CIDEC. The results confirmed that the mutant PPARG exhibited weaker transcriptional activation activity than the wild type (P<0.05). These findings likely explain the associations between the Asp7Gly substitution and the body measurements. Additionally, the Asp7Gly mutation may be used in molecular marker assisted selection (MAS) of cattle breeding in the future.
Streptococcus suis (S. suis) is a major pathogen in the pig industry and an important zoonotic agent that causes severe invasive diseases in humans. Previous studies based on multilocus sequence typing (MLST) and the associations between sequence types and genotypes or virulence suggested that North American S. suis serotype 2 isolates are composed of multiple populations. This study investigated the population structure of S. suis serotype 2 isolates in China. We constructed a phylogenetic tree for S. suis serotype 2 isolates based on 16S rRNA gene typing and MLST, studied associations between clades and sources, analyzed the genotype distributions of virulence markers [muramidase-released protein (MRP), extracellular protein factor (EF), and suilysin (SLY)] in different clades, computed the selection pressures for these virulence marker genes, and verified the associations between clades and virulence. There were two primary clades (populations) in the phylogenetic tree of S. suis serotype 2. The two populations were associated with different tissue tropisms. The genotypic distributions and selection pressures of MRP, EF, and SLY were different between the two populations, which suggested that they had different evolutionary paths. The two populations also displayed differences in virulence in experimentally infected mice. The results provide insights into the population structure of S. suis serotype 2 isolates in China and suggest that S. suis serotype 2 clade 1 is an overlooked population that deserves further evaluation.
Growth is under complex genetic control and uncovering the molecular mechanisms how the genes and polymorphisms affect economic growth traits, are important for successful marker-assisted selection and more efficient management strategies in commercial cattle populations. SIRT1 is a NAD(+)-dependent deacetylase that belongs to the class III histone deacetylases. It plays an important role in numerous fundamental cellular processes including gene silencing, DNA repair, and metabolic regulation. In addition, SIRT1 acts as an inhibitor of adipogenesis and has been associated with body weight regulation. The objective of the present study was to identify single nucleotide polymorphisms (SNPs) of bovine SIRT1 using 1255 animals representing the five main Chinese breeds and to determine if these SNPs are associated with economically important traits in Nanyang cattle. The approach consisted of resequencing SIRT1 using a panel of DNA from unrelated animals of five different breeds and the process revealed five novel SNPs. SNPs g.17324T>C and g.17491G>A exhibited a high degree of linkage disequilibrium in all tested breeds. Seven major haplotypes accounting for 91.2% of the alleles were observed and the haplotype GCCGA was the most common haplotype in NY, QC, LX and JX breeds. An association analysis was performed between the five SNPs and six performance traits. SNP g.-274C>G was demonstrated to have a strong effect on 24-months-old body weight and g.17379A>G polymorphism was related to 6 and 12-months-old body weight in NY population, although these effects did not remained significant after the Bonferroni correction. Our results provide evidence that polymorphisms in SIRT1 are associated with growth efficiency traits, and may be used for marker-assisted selection and management in feedlot cattle.
Deleted in breast cancer 1 (DBC1, KIAA1967, p30 DBC) is a novel protein that has been recently shown to bind and regulate SIRT1. Loss of function of DBC1 increased SIRT1 deacetylase activity, which promotes "browning" of WAT by deacetylating peroxisome proliferator-activated receptor (PPAR?) on Lys268 and Lys293. In the present study, we have cloned and characterized the bovine DBC1 gene. Two transcript variants of bovine DBC1 were identified, designated DBC1-A and DBC1-B, respectively, which were both located in nucleus. Protein sequence analysis showed that DBC1-A was well conserved across species. Expression analysis of DBC1 in seven different tissues of calves and bulls by RT-PCR indicated that the two transcripts were ubiquitously expressed. However, the relatively level of DBC1-A was higher when compared to DBC1-B in all examined tissues. Surprisingly, the expression of DBC1-A was extraordinary high in calves adipose tissue, which implicated its potential key role in regulating calve adipocyte development. These findings provide new insight into our understanding of the biochemical characteristics and physiological role of bovine DBC1.
Fibrogenic mesenchymal cells including fibroblasts and myofibroblasts play a key role in intestinal fibrosis, however, their precise role is largely unknown. To investigate their role in intestinal fibrosis, we analyzed the lesions of chronic colitis in C57BL/6 (B6) mice induced by dextran sulfate sodium (DSS). B6 mice exposed to single cycle administration of DSS for 5 days developed acute colitis that progressed to severe chronic inflammation with dense infiltrates of mononuclear cells, irregular epithelial structure, thickening of colonic wall, and persistent deposits of collagen. Increased mRNA expressions of proinflammatory cytokines are correlated with extensive cellular infiltration, and the mRNA expressions of collagen 1, transforming growth factor (TGF)-?, and matrix metalloproteinases were also enhanced in the colon. In the colon of chronic DSS colitis, fibroblasts (vimentin(+), ?-smooth muscle actin (?-SMA)(-)) were increased in both mucosal and submucosal layers, while myofibroblasts (vimentin(+), ?-SMA(+)) were increased in mucosal but not in submucosal layers. Primary mouse subcutaneous fibroblast cultures experiments revealed that exogenously added TGF-? 1 substantially augmented the expressions of both vimentin and ?-SMA proteins with increased production of collagen. In conclusion, profibrogenic mesenchymal cells play an important role in the development of intestinal fibrosis in this chronic DSS-induced colitis model.
Mast cells play a key role in the pathophysiology of inflammatory bowel disease (IBD). Tranilast, a mast cell stabilizer, has been empirically used for IBD in Japan, but its precise role in the treatment of IBD is largely unknown. To investigate the role of tranilast for the treatment of IBD, tranilast was administered intrarectally to mice with dextran sulfate sodium (DSS)-induced colitis. Tranilast ameliorated DSS colitis clinically and pathologically, as demonstrated by decreased number and degranulation of mast cells in the colon. mRNA expression was increased for tumor necrosis factor-alpha, interferon-gamma and interleukin (IL)-6, and decreased for IL-10 in the colon of DSS colitis mice. In contrast, tranilast markedly decreased expression of mRNAs for the pro-inflammatory cytokines, and increased that of the anti-inflammatory cytokines. Moreover, tranilast increased heme oxygenase (HO)-1 expression on colonic epithelial cells as well as on colon-infiltrating cells of DSS colitis. In conclusion, tranilast ameliorated DSS colitis by regulating mast cell degranulation, decreasing inflammatory cytokines and increasing anti-inflammatory cytokines. Tranilast might exert these effects partly through enhanced HO-1 expression in the colon, suggesting a potential adjunctive therapy for IBD.
Cardiac myosin is the most abundant and bioactive protein presented in heart tissue. In this work, a simple method was described for preparation of high purity cardiac myosin for further research. With the use of a domestic pig left ventricle as the starting tissue, this method yielded 4.5 mg myosin per gram of wet tissue with a specific activity of 0.293 U/mg. The purification factor was 4.9-fold with an activity recovery of 42.1%. In addition, the investigation of fluorescence spectrum shown that the emission intensity of cardiac myosin has a dramatic decrease under the influence of an acidic environment or at temperatures higher than 30 degrees C, which could influence the normal physiological function of cardiac myosin.
Understanding the mechanisms controlling transcription of a gene requires the identification and characterization of its cis-acting regulatory elements. A highly useful approach to the identification and characterization of cis-acting elements has been the systematic coupling of genomic fragments to reporter constructs, so called "promoter bashing". The expression from such reporters must be normalized for differences in transient transfection efficiency between cells and replicates. A novel dual color fluorescent reporter system to assay the promoter activity of a genomic DNA fragment of interest was established by cloning a Discosoma red fluorescent protein gene and a green fluorescent protein gene into a single vector, giving a system in which the ratio between red and green fluorescence is proportional to promoter activity. This system allows real time quantitative monitoring of promoter activity. We validated this approach by assaying the cis-acting regulatory potential of the peroxisome proliferators-activated receptor gamma2 gene.
Related JoVE Video
Journal of Visualized Experiments
What is Visualize?
JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.
How does it work?
We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.
Video X seems to be unrelated to Abstract Y...
In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.