Aberrant up-regulation of LAMB3 and LAMC2 by promoter demethylation in gastric cancer

The LAMB3 and LAMC2 genes encode the laminin-5 β3 and γ2 chains, respectively, which are parts of laminin-5, one of the major components of the basement membrane zone. Here, we report the frequent up-regulation of LAMB3 and LAMC2 by promoter demethylation in gastric cancer. Gene expression data analysis showed that LAMB3 and LAMC2 were up-regulated in various tumor tissues. Combined analyses of DNA methylation and gene expression of both genes in gastric cancer cell lines and tissues showed that DNA hypomethylation was associated with the up-regulation of both genes. Treatment with a methylation inhibitor induced LAMB3 and LAMC2 expression in gastric cancer cell lines in which both genes were silenced. By chromatin immunoprecipitation assay, we showed the activation histone mark H3K4me3 was associated with the expression of both genes. The expression level of LAMB3 affected multiple malignant phenotypes in gastric cancer cell lines. These results suggest that epigenetic activation of LAMB3 and LAMC2 may play an important role in gastric carcinogenesis.     

Lipopolysaccharide-induced hepatic oxidative injury is not potentiated by knockout of GPX1 and SOD1 in mice

Knockout of copper, zinc-superoxide dismutase (SOD1) and (or) cellular glutathione peroxidase (GPX1) has been reported to have dual impacts on coping with free radical-induced oxidative injury. Because bacterial endotoxin lipopolysaccharide (LPS) triggers inflammatory responses involving the release of cytokines, nitric oxide and superoxide in targeted organs such as liver, in this study we used SOD1 knockout (SOD1−/−), GPX1 knockout (GPX1−/−), GPX1 and SOD1 double-knockout (DKO) and their wild-type (WT) mice to investigate the role of these two antioxidant enzymes in LPS-induced oxidative injury in liver. Mice of the four genotypes (2 month old) were killed at 0, 3, 6 or 12 h after an i.p. injection of saline or 5 mg LPS/kg body weight. The LPS injection caused similar increase in plasma alanine aminotransferase among the four genotypes. Hepatic total glutathione (GSH) was decreased (P < 0.05) compared with the initial values by the LPS injection at all time points in the WT mice, but only at 6 and 12 h in the other three genotypes. The GSH level in the DKO mice was higher (P < 0.05) than in the WT at 6 h. Although the LPS injection resulted in substantial increases in plasma NO in a time-dependent manner in all genotypes, the NO level in the DKO mice was lower (P < 0.05) at 3, 6 and 12 h than in the WT. The level in the GPX1−/− and SOD1−/− mice was also lower (P < 0.05) than in the WT at 3 h. The LPS-mediated hepatic protein nitration was detected in the WT and GPX1−/− mice at 3, 6 or 12 h, but not in the SOD1−/−. In conclusion, knockout of SOD1 and (or) GPX1 did not potentiate the LPS-induced liver injury, but delayed the induced hepatic GSH depletion and plasma NO production.     

Effect of recombinant growth hormone on expression of growth hormone receptor,insulin-like growth factor mRNA and serum level of leptin in growing pigs

Sixteen Large White × Landrace castrated male pigs were allotted into treatment and control group. The treatment group was injected intramuscularly with recombinant porcine growth hormone (rpGH, 4 mg d⁻¹) and the control group with vehicle for 28 days. Animals were slaughtered 4 h after final injection for liver, longissimus dorsi (LD) muscle and blood sampling. Serum concentration of insulin-like growth factor 1 (IGF-I) and leptin were determined by RIA. The total RNA was extracted from tissues to measure the abundance of growth hormone receptor (GHR), IGF-I mRNA by RT-PCR with 18S rRNA internal standard. Results showed that rpGH enhanced the average daily weight gain by 26.1% (P < 0.05), the serum IGF-I concentration by 70.94% (P < 0.01), decreased serum leptin by 34.8% (P < 0.01). The relative abundance of GHR and IGF-I mRNA in liver were increased by 24.45% (P < 0.05) and 45.30% (P < 0.01), respectively, but no difference of GHR (P > 0.05) and IGF-I mRNA (P > 0.05) in LD between GH treated and control group was found. These results suggest that rpGH can up-regulate hepatic GHR and IGF-I gene expression and improve animal growth. However the effect of rpGH on GHR and IGF-I gene expression are tissue-specific.     

Molecular characterization and polymorphisms of the caprine Somatostatin (<Emphasis Type="Italic">SST</Emphasis>) and SST Receptor 1 (<Emphasis Type="Italic">SSTR1</Emphasis>) genes that are linked with growth traits

Somatostatin (SST) and its receptors (SSTR1-5) appear to be important in central regulation of many metabolic systems that affect growth, adiposity and nutrient absorption. In this study, we investigated polymorphisms within the caprine SST and SSTR1 genes and determined their relationship with growth traits. As there were no sequence information of the caprine SST and SSTR1 genes, we explored their DNA sequence and genomic organizations. The caprine SST gene is organized in two exons and is transcribed into an mRNA containing 351 bp of sequence coding for a protein of 116 amino acids. Its protein sequences showed substantial similarity (97–99%) to its respective orthologs from cattle, human and mouse. We also cloned and sequenced a 1.2 kb DNA fragment which contained the major part of the coding region and 3′ UTR of the caprine SSTR1 gene. We then detected the polymorphisms in these determined sequences by PCR-SSCP and DNA sequencing methods in 459 goats from four breeds. Four SNPs (GU014693:g.647T>C, GU014693:g.844A>C, GU014693:g.970T>C, GU014693:g.1039T>A), segregating as two haplotypes (T-A-T-T and C-C-C-A), were identified in intron 1 of the caprine SST gene and showed the associations to body length and body height (P < 0.05). Two SNPs (GU014695:g.801 C>T, GU014695:g.948 C>T) were identified in the caprine SSTR1 gene. Significant associations between the three genotypes of GU014695:801 C>T and body length, body height, and chest circumference was observed (P < 0.05). These results suggest that the caprine SST and SSTR1 genes are strong candidate genes that influence growth traits in goat.     

Investigating the heat tolerance and production performance in local chicken breed having normal and dwarf size

Heat stress significantly impairs the growth performance of broilers, which causes serious losses to the poultry industry every year. Thus, understanding the performance of indigenous chicken breeds under such environment is crucial to address heat stress problem. The purpose of this study was to investigate the effects of heat stress (HS) on production performance, tissue histology, heat shock response (HSP70, HSP90), and muscle growth-related genes (GHR, IGF-1, and IGF-1R) of Normal yellow chicken (NYC) and Dwarf yellow chicken (DYC). Seventy-two female birds from each strain were raised under normal environmental conditions up to 84 days, with birds from each strain being divided into two groups (HS and control). In the HS group, birds were subjected to high temperature at 35 ± 1 °C for 8 h daily and lasted for a week, while in the control group, birds were raised at 28 ± 1 °C. At 91 days old, bird's liver, hypothalamus, and breast muscle tissues were collected to evaluate the gene expression, histological changes, and the production performance. The Feed intake, weight gain ratio, total protein intake and protein efficiency ratio showed a significant reduction in the treatments (P < 0.01) and treatment × strain interaction (P < 0.05) with breast muscle rate significantly reducing among the treatments (P < 0.01) after 7 days of HS. Correspondingly, total abdominal fat showed significant change among treatment and strain (P < 0.01, P < 0.05), respectively. Besides, HS markedly upregulated the mRNA expression of HSP70 and HSP90 in the pectoralis major of both chicken strains, but no significant increase (P < 0.05) was found in mRNA expression of HSP90 in liver and hypothalamus tissues of both chicken strains. Moreover, HS significantly upregulated (P < 0.05) the expression of lipogenic genes (FASN, ACC) in liver tissues of NYC, while mRNA expression of these genes showed no variation in DYC. Similarly, HS downregulated the mRNA expression of muscle growth-related genes (GHR, IGF-1, and IGF-1R). Consequently, the histopathological analysis showed that histological changes were accompanied by inflammatory cell infiltration in liver tissues of both chicken strains; however, histopathological changes were more severe in NYC than dwarf chicken strain. Conclusively, this study depicted that the production performance and growth rate varied significantly between treatment and control group of NYC. However, heat treatment in DYC has not shown significant damaging consequences as compared to the control group that signifies the vital role of the dwarf trait in thermal tolerance.     

不同产地浙贝母生物碱含量及其合成相关基因表达研究

为了探究浙贝母(Fritillaria thunbergii)药效成分积累量与生物碱合成相关基因表达水平之间的关系,该研究采用UPLC-MS、qPCR技术分别测定不同产地11个样品中总生物碱(贝母素甲和贝母素乙之和)含量和3个参与生物碱合成途径相关基因(HMGR、FPS和DXR)的表达量,同时运用生物统计学方法分析成熟期鳞茎生物碱含量与各基因表达量之间的相关性。结果表明:不同产地浙贝母成熟期鳞茎总生物碱含量存在显著差异(P<0.05),为0.2105% ～ 0.4612%; HMGR和FPS基因在盛花期组织表达、盛花期至成熟期鳞茎表达变化趋势同生物碱含量变化趋势基本一致; DXR基因在成熟期鳞茎中表达量最高,盛花期组织表达、盛花期至成熟期鳞茎表达变化趋势同生物碱含量变化趋势大体不一致; HMGR、FPS基因表达量分别与贝母素甲、贝母素乙和总生物碱含量呈显著或极显著正相关性(P<0.05或P<0.01),以FPS基因表达量与生物碱含量相关系数为最高,相关系数分别为0.672,0.631,0.664,DXR基因与生物碱含量呈低度相关性。由此可以推断,生物碱的积累受MVA途径中HMGR、FPS基因协同调控或者修饰作用明显,受MEP途径中DXR基因调控作用不明显。     

Characterization of the expression profiles of calpastatin (CAST) gene in chicken

The calpain system, a Ca²⁺-activated protease family, plays an important role in postmortem tenderization of skeletal muscle due to its involvement in the degradation of important myofibrillar and associated proteins, as well as in cytoskeletal remodeling and regulation of muscle growth. In this study, we quantified the expression of calpastatin (CAST) in two Chinese chicken breeds (mountainous black-bone chicken breed (MB) and a commercial meat type chicken breed (S01)), to discern the tissue and age-related specific expression pattern and its potential role on muscle tissue metabolism. Real-time quantitative PCR (RT-qPCR) assay was developed for accurate measurement of CAST mRNA levels in various tissues from chicken with different ages (0, 2, 4, 6, 8, 10, and 12 week). CAST mRNA was detected in collected organs. The heart and leg muscle tissues had the highest expression of CAST than other tissues from the same chicken (P < 0.01). Age-related expression pattern of CAST gene was evident in breast muscle, liver, and brain tissues (P < 0.05), but not in heart and leg muscle tissues (P > 0.05). Overall, the CAST mRNA level exhibited a “rise-decline-rise-decline” developmental change in breast muscle and liver, with the highest expression at 2 weeks and the lowest expression at 8 weeks. The S01 chicken had significantly higher expression of CAST in breast muscle and heart than the MB chicken (P < 0.05) at 10 weeks. Our results suggested the CAST expression may be related to muscle fiber development.     

Fluoxetine, 17-β estradiol or folic acid combined with intra-lateral septal infusions of neuropeptide Y produced antidepressant-like actions in ovariectomized rats forced to swim

Folic acid is antidepressant, either alone or combined with several antidepressant drugs. However, the antidepressant-like actions of folic acid combined with intra-lateral septal (LSN) infusions of neuropeptide Y (NPY) in the forced swimming test (FST) have not been tested before. Thus, systemic injections of fluoxetine (20.0 mg/kg, P < 0.05; s.c.) or 17-β estradiol (10.0 μg/rat, P < 0.05; s.c.) or oral administrations of folic acid (50.0 mg/kg, P < 0.05; 75.0 mg/kg, P < 0.05) or NPY intra-LSN (3.0 μg, P < 0.05; 3.5 μg, P < 0.05) reduced immobility of ovariectomized Wistar rats. Subthreshold doses of: folic acid (25.0 mg/kg) or 17-β estradiol (5.0 μg/rat, P < 0.05) or fluoxetine (15.0 mg/kg, P < 0.05; s.c.) combined with subthreshold doses of NPY (2.5 μg/rat, P < 0.05; intra-LSN) and these combinations produced antidepressant-like actions; which were canceled by BIBP 3226 (a NPY-Y1 receptor antagonist). It is concluded that folic acid produced antidepressant-like effects probably through the participation of the NPY Y1 receptors found in the lateral septal nuclei.     

Effects of chromium picolinate supplementation on the growth,carcass quality and gene expression of beef during the finishing period

Four groups of 12 young beef, as similar as possible with respect to age and weight, were fed a basic diet. The addition fed to four groups was 0, 200, 600, and 1,200 mg of organic chromium (chromium picolinate CrPic) per kg concentrated feed. The results showed that there was no effect on overall growth performance and dressing percentage and pure meat percentage when adding different CrPic content, but significant difference was found between 0 group and other three groups in percentage of high grade cuts (P < 0.05). The Cr content in Kidney, Musculus diaphragm, Semitendinosus muscles and Longissimusdorsi tissues have no difference in four groups (P > 0.05), but there was significant difference in liver tissues between 0, 200, 600 groups and 1200 group (P < 0.05). Gene expression analysis indicated that there were no differences in five genes expression in liver and muscle tissues in four groups (P > 0.05), but mRNA expression amount of FOX1, FSTL and MATR3 always had same trends whatever in liver or muscle tissues. However the RPLOP gene expression amount has large difference between liver and muscle.     

脑红蛋白在赛加羚羊主要内脏器官中的表达与定位

赛加羚羊(Saiga tatarica)属于我国一级重点保护野生动物,其原产地主要为高寒低氧地区,现存种群则主要栖息于中亚地区的荒漠及半荒漠草原上。脑红蛋白是一种存在于脊椎动物体内具有运输和储存血氧能力的球蛋白,在动物适应低氧过程中具有重要的生理功能。为了初步探究赛加羚羊对低氧环境的耐受性机制,运用免疫组织化学染色法与实时荧光定量PCR技术,对脑红蛋白及脑红蛋白基因(NGB)在赛加羚羊的心、肝、脾、肺、肾等5种主要内脏器官中的分布规律与表达情况进行了探究。免疫组织化学染色结果显示,脑红蛋白在赛加羚羊的心、肝、脾、肺、肾中均有分布,阳性表达主要分布在其心肌细胞、肝细胞、脾白髓区中的淋巴细胞、肺泡细胞以及肾小球内皮细胞。实时荧光定量PCR结果显示,脑红蛋白基因在赛加羚羊心、肝、脾、肺、肾中的表达量不同,脾的表达量最高,心的表达量次之,两者均显著高于肝、肺和肾(P < 0.05);其后依次为肝、肾、肺,其中,肝的表达量显著高于肾和肺(P < 0.05),肾和肺之间表达量差异不显著(P > 0.05),肺的表达量最低。上述研究表明,脑红蛋白在赛加羚羊的主要内脏器官中均有阳性表达,不同内脏器官中的表达量不同,这表明脑红蛋白可能参与了这些内脏器官的氧利用过程,具体机制有待进一步探讨。     

Liver dominant expression of fatty acid synthase (FAS) gene in two chicken breeds during intramuscular-fat development

Fatty acid synthase (FAS) is a key enzyme of lipogenesis. In this study, the FAS mRNA expression patterns were examined in three fat related tissues (liver, breast and thigh) at different developmental stages in two chicken breeds (Beijing-You, BJY and Arbor Acres broiler, AA). Results of the Real time-qPCR showed that the expression of FAS mRNA level in liver was significantly higher (P < 0.01) than that in breast and thigh in both two chicken breeds. Significant differences of FAS mRNA expression in liver were found between BJY and AA chickens during different developmental stages. After the contents of intramuscular-fat (IMF) and the liver fat were measured, the correlation analysis was performed. In liver, the FAS mRNA level was highly correlated with hepatic fat content (r = 0.891, P < 0.01 for BJY; r = 0.901, P < 0.01 for AA). On the contrary, the FAS expression level in both breast and thigh tissues were relatively low, stable and there was no correlation between the FAS mRNA level and IMF content in breast and thigh tissues of each breed. The results here can contribute to the knowledge on the developmental expression pattern of FAS mRNA and facilitate the further research on the molecular mechanism underlying IMF deposition in chicken.     

Caecal microbiota could effectively increase chicken growth performance by regulating fat metabolism

It has been established that gut microbiota influences chicken growth performance and fat metabolism. However, whether gut microbiota affects chicken growth performance by regulating fat metabolism remains unclear. Therefore, seven-week-old chickens with high or low body weight were used in the present study. There were significant differences in body weight, breast and leg muscle indices, and cross-sectional area of muscle cells, suggesting different growth performance. The relative abundance of gut microbiota in the caecal contents at the genus level was compared by 16S rRNA gene sequencing. The results of LEfSe indicated that high body weight chickens contained Microbacterium and Sphingomonas more abundantly (P < 0.05). In contrast, low body weight chickens contained Slackia more abundantly (P < 0.05). The results of H & E, qPCR, IHC, WB and blood analysis suggested significantly different fat metabolism level in serum, liver, abdominal adipose, breast and leg muscles between high and low body weight chickens. Spearman correlation analysis revealed that fat metabolism positively correlated with the relative abundance of Microbacterium and Sphingomonas while negatively correlated with the abundance of Slackia. Furthermore, faecal microbiota transplantation was performed, which verified that transferring faecal microbiota from adult chickens with high body weight into one-day-old chickens improved growth performance and fat metabolism in liver by remodelling the gut microbiota. Overall, these results suggested that gut microbiota could affect chicken growth performance by regulating fat metabolism.     

Trypanosoma evansi: Activities of adenine nucleotide degradation enzymes in cerebral cortex of infected rats

This study aimed to evaluate the activities of the ectoenzymes NTPDase and 5′-nucleotidase in synaptosomes from cerebral cortex of rats experimentally infected with Trypanosoma evansi. The animals were divided in four groups (n = 10) according to the time and degree of parasitemia (groups A, B, C and D). The animals from group A were euthanized on day 3 (low parasitemia), group B on day 5 (high parasitemia) and group C on day 15 (low parasitemia). Group D consisted of healthy rats (not-infected, n = 15) and were divided in three periods (n = 5) in order to compare with the infected groups. After euthanasia, cerebral cortex was removed for the preparation of synaptosomes and enzymatic assays. Group A showed no changes in enzymatic activities compared with control. The hydrolysis of ATP, ADP and AMP by the enzymes NTPDase and 5′-nucleotidase were increased (P < 0.05) in group B (38%, 140% and 61%, respectively) when compared with control. In the group C it was observed a decreased (22%) hydrolysis of ATP when compared with control group. The activities of NTPDase and 5′-nucleotidase in synaptosomes alters the acute phase of the disease when the number of circulating parasites is high, thus the change observed is probably due to the parasitemia.     

三月竹开花前后几种重要代谢物差异研究

三月竹(Chimonobambusa opienensis)为禾本科竹亚科寒竹属植物,是国家一级保护动物大熊猫主食竹种,也是当地农民和笋竹加工企业的一大重要经济来源。该研究以开花与未开花三月竹为材料,采用吸光光度法、差量法、苯酚硫酸法、考马斯亮蓝G-250染色法、愈创木酚显色法、邻苯三酚自氧化法,对三月竹开花前后各部位多种重要代谢物进行测定,并用统计方法进行差异分析。结果表明:三月竹开花后,叶与次生枝叶绿素含量分别下降14.42%和71.39%(P0.05);次生枝与主枝油脂含量分别下降20.93%和26.04%(P0.05);叶与次生枝可溶性糖含量分别上升21.04%和17.81%,淀粉上升8.33%和8.21%,纤维素上升17.62%和8.52%(P0.05);叶与次生枝可溶性蛋白质含量分别下降30.07%和37.31%(P0.05);上叶POD活性上升122.01%(P0.05)。这说明三月竹开花与多种有机营养和生化指标的变化关联,可通过监测叶与次生枝叶绿素含量,次生枝与主枝油脂含量,叶与次生枝可溶性糖、淀粉、纤维素含量,叶与次生枝可溶性蛋白质含量,上叶POD活性来实现对三月竹生长的管理调控。该研究结果可为进一步研究三月竹开花衰老机理和延期开花提供参考。     

Expression profiling of candidate genes for abdominal fat mass in domestic chicken <Emphasis Type="Italic">Gallus gallus</Emphasis>

The quantitative traits of mass and percentage of abdominal fat in chicken and various types of obesity in mammals are homologous and functionally similar. Therefore, the genes involved in obesity development in humans and laboratory rodents as well as those responsible for pig lard thickness could be involved in abdominal fat deposition in broilers. Expression of candidate genes FABP1, FABP2, FABP3, HMGA1, MC4R, PPARG, PPARGC1A, POMC and PTPN1 was studied in fat, liver, colon, muscle, pituitary gland, and brain in chicken (broilers) using real-time PCR. Significant difference in the HMGA1 gene expression in the liver of broiler chicken with high (3.5 ± 0.18%) and low (1.9 ± 0.56%) abdominal fat concentration has been revealed. The expression of this gene was been shown to correlate with the amount (0.7, P ≤ 0.01) and mass (0.7, P ≤ 0.01) of abdominal fat. The PPARG gene expression in liver in the same chicken subsets was also significantly different. Correlation coefficients of the gene expression with the abdominal fat amount and mass were respectively 0.55 (P ≤ 0.05) and 0.57 (P ≤ 0.01). Based on these results, we suggest that the HMGA1 and PPARG genes are involved in abdominal fat deposition. The search for single nucleotide polymorphisms (SNPs) in the HMGA and PPARG regulatory regions could facilitate identifying genetic markers for broiler breeding according to the mass and percentage of abdominal fat.     

Differential allelic expression of c.1568C > A at UGT2B15 is due to variation in a novel cis-regulatory element in the 3'UTR

Differential allelic expression (DAE) is a powerful tool to identify cis-regulatory elements for gene expression. The UDP-glucuronosyltransferase 2 family, polypeptide B15 (UGT2B15), is an important enzyme involved in the metabolism of multiple endobiotics and xenobiotics. In the present study, we measured the relative expression of two alleles at SNP c.1568C>A (rs4148269) in this gene, which causes an amino acid substitution (T523K). An excess of the C over the A allele was consistently observed in both liver (P = 0.0021) and breast (P = 0.012) samples, suggesting that SNP(s) in strong linkage disequilibrium (LD) with c.1568C>A can regulate UGT2B15 expression in both tissues. By resequencing, one such SNP, c.1761T>C (rs3100) in 3′ untranslated region (UTR), was identified. Reporter gene assays showed that the 1761T allele results in a significantly higher gene expression level than the 1761C allele in HepG2, MCF-7, LNCaP, and Caco-2 cell lines (all P < 0.001), thus indicating that this variation can regulate UGT2B15 gene expression in liver, breast, colon, and prostate tissues. Considering its location, we postulated that this SNP is within an unknown microRNA binding site and can influence microRNA targeting. Considering the importance of UGT2B15 in metabolism, we proposed that this SNP might contribute to multiple cancer risk and variability in drug response.