黄芪多糖分级组分对鸡脾淋巴细胞增殖影响

目的：探讨黄芪多糖分级组分对鸡脾淋巴细胞增殖的影响。方法：将黄芪多糖经过不同浓度的乙醇进行分级沉淀,得到3个分级组分：A1、A2、A3;将320只鸡随机分为8组,分别皮下注射A1、A2、A3、A1＋A2、A1＋A3、A2＋A3、A1＋A2＋A3和蒸馏水,每隔7d各组随机捕杀鸡4只,培养脾淋巴细胞,NTT法测定黄芪多糖分级组分对鸡脾淋巴细胞增殖的影响。结果：A2、A3、A2＋A3、A1＋A2＋A3能显著促进淋巴细胞增殖。结论：黄芪多糖免疫功能的有效部位为A2、A3,尤以A2作用显著。     

鸡脂滴包被蛋白基因内含子6多态性与胴体及脂肪性状的相关性研究

试验采用PCR-RFLP方法检测脂滴包被蛋白(perilipin,PLIN)基因内含子6在济宁百日鸡、莱芜黑鸡等4个地方鸡种和1个培育品系中的遗传多态性,分析了多态位点不同基因型与鸡胴体及脂肪性状的相关性。结果发现,在5个供试群体中检测到1个多态位点,测序证实为新发现的鸡PLIN基因2 467 bp处G→A突变,该突变位点在供试群体中检测到3种基因型:A1A1、A1A2和A2A2,2个等位基因:A1和A2。等位基因A1在所有供试群体中均表现为优势等位基因。关联分析结果表明,PLIN基因2 467 bp位点对鸡部分胴体性状和脂肪性状影响显著(P<0.05)。多重比较结果表明,A1A1基因型个体活体重、屠体重、全净膛重、腹脂重和腹脂率均显著高于A2A2基因型个体(P<0.05)。A1A2基因型个体的胸肌肌内脂肪含量显著高于A1A1和A2A2基因型个体(P<0.05)。研究结果表明,PLIN基因对鸡胴体及脂肪性状有一定影响。     

Polymorphism in PLIN Gene Intron 6 and its Association with Carcass and Fatness Traits in Chicken

The genetic polymorphisms in PLIN gene were detected by PCR-RFLP method in four Chinese local chickens including Jining Bairi chicken,Laiwu Black chicken et al.,and one breeding strain.The correlations between the SNP and the carcass and fatness traits were analyzed.As a result,a novel G→A mutation at 2 467 bp in PLIN gene was identified in the five populations.Three genotypes (A1A1,A1A2 and A2A2) were detected,and allele A1 was predominant in all the five experimental populations.The statistical analysis showed that the 2 467 bp polymorphism locus was significant association with some carcass and fatness traits (P<0.05).The living body weight,carcass weight,eviscerated weight,abdominal fat weight and percentage of abdominal fat of A1A1 genotype was higher than A2A2 genotype in chickens (P<0.05).The breast intramuscular fat of A1A2 genotype was higher than that of A1A1 and A2A2 genotypes in chickens (P<0.05).The results showed that PLIN gene had effect on carcass and fatness traits in chickens.     

The genes of all seven CYP3A isoenzymes identified in the equine genome are expressed in the airways of horses

In the present study, we examined the gene expression of cytochrome P450 3A (CYP3A) isoenzymes in the tracheal and bronchial mucosa and in the lung of equines using TaqMan probes. The results show that all seven CYP3A isoforms identified in the equine genome, that is, CYP3A89, CYP3A93, CYP3A94, CYP3A95, CYP3A96, CYP3A97 and CYP3A129, are expressed in the airways of the investigated horses. Though in previous studies, CYP3A129 was found to be absent in equine intestinal mucosa and liver, this CYP3A isoform is expressed in the airways of horses. The gene expression of the CYP3A isoenzymes varied considerably between the individual horses studied. However, in most of the horses CYP3A89, CYP3A93, CYP3A96, CYP3A97 and CYP3A129 were expressed to a high extent, while CYP3A94 and CYP3A95 were expressed to a low extent in the different parts of the airways. The CYP3A isoenzymes present in the airways may play a role in the metabolic degradation of inhaled xenobiotics. In some instances, the metabolism may, however, result in bioactivation of the xenobiotics and subsequent tissue injury.     

aroA gene PCR-RFLP diversity patterns in Haemophilus parasuis and Actinobacillus species

The Haemophilus parasuis aroA gene encodes 5-enolpyruvylshikimate-3-phosphate synthase and participates in the aromatic amino acids and the folic acid universal metabolic pathway of bacteria. The application of aroA-based PCR-RFLP methodology yields a significant degree of diversity in H. parasuis and Actinobacillus species. PCR amplification of the aroA gene rendered a 1,067-bp fragment in all 15 H. parasuis serovars, and also in Actinobacillus pleuropneumoniae serotypes 1-12, Actinobacillus lignieresii, Actinobacillus equuli, Actinobacillus porcinus, Actinobacillus rossii, Actinobacillus suis, Actinobacillus ureae, Actinobacillus minor and Actinobacillus indolicus. Sau3AI and RsaI digestions of the aroA PCR products rendered seven different restriction fragment length polymorphism (RFLP) patterns: group I (H. parasuis serovars 1, 2, 4-6, and 8-15, A. porcinus and A. ureae), group II (H. parasuis serovars 3 and 7, and A. pleuropneumoniae serotypes 1, 4, 5, 9, 11 and 12), group III (A. lignieresii), group IV (A. pleuropneumoniae serotype 7), group V (A. pleuropneumoniae serotypes 2, 3, 6 and 8, A. equuli, A. rossii, A. minor and A. indolicus), group VI (A. suis) and group VII (A. pleuropneumoniae serotype 10). This is the first report describing the presence of aroA gene in H. parasuis, A. lignieresii, A. porcinus, A. rossii, A. suis, A. ureae, A. minor and A. indolicus and the data presented here demonstrates a significant degree of aroA genetic diversity in H. parasuis and species of the genus Actinobacillus.     

黄芪多糖分级组分对鸡疫苗免疫效果的影响

目的探讨黄芪多糖分级组分对禽流感—新城疫重组二联活疫苗免疫效果的影响。方法将黄芪多糖经过不同浓度的乙醇进行分级沉淀,得到三个分级组分:A1、A2、A3;将320只鸡随机分为8组,分别皮下注射A1、A2、A3、A1+A2,A1+A3、A2+A3、A1+A2+A3和蒸馏水,首免后每隔7天采血检测疫苗抗体效价。结果A2、A3能显著提高疫苗抗体效价。结论黄芪多糖免疫功能的有效部位为A2、A3,尤以A2作用显著。     

Cloning and Bioinformatics Analysis of 4 Alternative Splice Variants of MEF2A Gene in Mashen Pig

This study was aimed to clone the different splicing variants of myocyte enhancer factor 2A (MEF2A) gene in Mashen pig. According to the prediction results of the alternative splicing in the RNA-Seq sequencing, the coding region of MEF2A gene exons 5-8 were amplified. We analyzed the sequence characteristics of different splicing variants of MEF2A gene in Mashen pig, predicted conservative structure domain, constructed phylogenetic tree and compared the homology of MEF2A amino acid in different species by bioinformatics softwares. The results showed that 4 alternative splice variants of MEF2A gene were obtained. The exons 5-8 of MEF2A1 spliced normally. MEF2A2 lacked the exon 5, and the 5'end of exon 6 was 138 bp longer, the 3'end of exon 7 was 102 bp longer. MEF2A3 lacked exon 5, and the 5'end of exon 6 was 138 bp longer. The 3'end of MEF2A4 exon 7 of was 102 bp longer. The protein, encoded by inserted 138 bp sequence, contained a conserved domain-HJURP_C, which was the result of MEF2A participating in hepatocyte fibrosis. The MEF2A1, MEF2A4 and MEF2A1 of pig submitted in GenBank (accession No.NP_001090890.1) belonged to a subgroup, homology up to 98.9%. The MEF2A2, MEF2A3 and MEF2A2 of pig submitted in GenBank (accession No.NP_001093168.1) belonged to a subgroup, the homology was 98.2% and 98.9%, respectively. The successful cloning of 4 alternative splice variants of MEF2A gene laid the foundation for further study on the function of MEF2A protein.     

祁连山自然保护区高寒草地高等真菌名录初报(一)

调查采集和报道了自祁连山自然保护区高寒草地的蘑菇科真菌21种。其中,粗柄蘑菇(Agaricus spissicaulis)、赭鳞蘑菇(A.subrufescens)、球基蘑菇(A.abruptibulbus)、褐缘鳞蘑菇(A.squamuliferus)、短柄蘑菇(A.ingratus)、赭褐蘑菇(A.langei)、紫褐蘑菇(A.rubellus)、污白蘑菇(A.excellens)、橙黄蘑菇(A.perrarus)、菌索蘑菇(A.lamnipes)、细环柄菇(Lepiota clypeolaria)等11种为甘肃省新纪录种。对其生长的生态环境和经济价值及在草地物质循环中的作用等进行了介绍。     

马身猪MEF2A基因4种可变剪接体的克隆和生物信息学分析

本研究旨在克隆马身猪肌细胞增强因子2A（myocyte enhancer factor 2A,MEF2A）基因的不同剪接体类型。依据转录组测序对可变剪接的预测结果,试验扩增MEF2A基因第5~8外显子之间的编码区,用生物信息学软件分析了马身猪MEF2A基因不同剪接体的序列特性,预测保守结构域,构建系统进化树,比对不同物种MEF2A的氨基酸同源性。结果显示,获得了4个MEF2A基因的剪接体,MEF2A1的第5~8外显子正常剪接;MEF2A2缺失了第5外显子,第6外显子的5'端多出138 bp,第7外显子的3'端多出102 bp;MEF2A3缺失了第5外显子,第6外显子的5'端多出138 bp;MEF2A4第7外显子的3'端多出102 bp。插入的138 bp序列编码的蛋白质中存在1个保守结构域HJURP_C,这可能与MEF2A参与肝细胞纤维化作用有关。MEF2A1和MEF2A4与猪MEF2A1（GenBank登录号:NP_001090890.1）同属于一个亚群,同源性高达98.9%,MEF2A2和MEF2A3与猪MEF2A2（GenBank登录号:NP_001093168.1）同属于一个亚群,同源性分别为98.2%和98.9%。本试验成功克隆了4个MEF2A基因的剪接体,为进一步研究其蛋白功能奠定基础。     

Genetic polymorphism of bovine beta-casein gene in Japanese dairy farm herds

The aim of this study was to investigate beta-casein polymorphism among 320 Japanese cows sampled from eight dairy farms. We used a newly-developed genotyping method that involved collecting DNA from hairs and a Cycleave polymerase chain reaction (PCR) assay to detect the A1, A2, and B variants. Results revealed the presence of five genotypes (A1A1, A2A2, A1A2, A1B, and A2B). We found that the most common genotype was A2A2 (0.42), followed by A1A2 (0.39) and A1A1 (0.11). The A1B and A2B genotypes were less frequent (<0.05). The frequencies of alleles A1, A2, and B were calculated to be 0.32, 0.64, and 0.04, respectively. Our study is the first to show the current status of beta-casein polymorphisms in Japanese dairy farms. Given the adverse effects of A1 beta-casein on human health, attempts have been made to develop herds consisting solely of A2A2 cows. Our study provides a reference for improving cow populations in Japanese dairy farms. The Cycleave PCR-based assay we developed here can be used for rapid and reliable genotyping of bovine beta-casein.     

九龙藏黄牛和九龙牦牛β-酪蛋白基因型分析

为比较九龙藏黄牛和九龙牦牛β-酪蛋白（β-CN）的遗传变异体,试验采用酸性尿素聚丙烯酰胺凝胶电泳分析了九龙藏黄牛（n=42）和九龙牦牛（n=17）β-CN的基因型。结果表明,在九龙藏黄牛、九龙牦牛的β-CN中共检测到4 种等位基因,包括A1、A2、B、C,其中在九龙藏黄牛中有7 种基因型：A1A1、A1A2、BB、A1B、A2B、A1C、BC,优势等位基因为A1（频率0.702 4）,优势基因型为A1A1（频率0.547 6）;在九龙牦牛样本中有2 种基因型：A1A2、A2A2,优势等位基因为A2（频率0.764 7 ）。试验表明,九龙藏黄牛与九龙牦牛β-CN均表现出多态性,但优势等位基因明显不同。     

磷酸二酯酶基因在小型猪骨骼肌组织的表达

提取中国实验用小型猪骨骼肌组织总RNA，应用反转录聚合酶链反应测定磷酸二酯酶(PDE)同工酶在骨骼肌组织中的表达分布。结果可见PDE1A、1C、2A、3A、3B、4A、4B、4C、4D、5A、7A、7B、8A、8B、9A和11A共16种PDE同工酶mRNA在中国实验用小型猪骨骼肌组织中表达，其中PDE1A、1C、2A、3A、4B、4C、8A和8B共8种在人和其他动物骨骼肌组织中的表达分布未见报道，PDE1B和10A3种同工酶未见表达。16种PDE同工酶mRNA在骨骼肌组织中的表达，从电泳条带上可以看到各种PDE表达量存在着差异，但需要进一步做定量分析加以评定。     

细胞色素P4503A酶与双峰驼物质代谢的特性

细胞色素P4503A(CYP3A)是细胞色素P450(CYP)酶超家族在动物肝脏中最主要的功能形式,约占肝脏总CYP酶含量的30%,参与45%~60%常用药物在体内的代谢转化过程。人们对各种CYP3A酶开展较为系统的研究,包括CYP3A酶分子结构、遗传多样性、体内外活性及其影响因素等。论文从CYP3A亚家族的生物学特性、对内源性化合物和外源性化学物质代谢的意义、CYP3A基因的表达与调控、CYP3A酶的诱导与抑制及双峰驼CYP3A酶研究进展等方面加以综述。     

幼龄鸡卵黄囊中^3H—VA的吸收代谢及其利用的研究

选用出壳后１２时龄的ＡＡ肉雏１４５只，采用放射性同位素＾Ｈ示踪方法，分别进行了饲养试验，屠补试验和代射试验研究表明，在全价日粮和无ＶＡ纯日粮条件下，雏鸡卵黄囊中的内源性３＾Ｈ－ＶＡ在体内发挥持续时间分别为２１天和１４天。卵黄囊（内源）３＾Ｈ－ＶＡ在鸡肠道中的吸收代谢，与饲料（外源）中营养物质相比，有着独自的特点。卵黄囊中内源３＾Ｈ－ＶＡ在雏鸡肠料中外源ＶＡ之间存在有动态交换关系，内源３＾Ｈ－ＶＡ主     

黄芪多糖分级组分对鸡血清中IL-2含量的影响

本试验旨在探讨黄芪多糖分级组分对鸡血清中IL-2含量的影响。将黄芪多糖经过不同浓度的乙醇进行分级沉淀,得到3个分级组分:A1、A2、A3;将320只鸡随机分为8组,分别皮下注射A1、A2、A3、A1+A2,A1+A3、A2+A3、A1+A2+A3和蒸馏水,每隔7 d采血检测血清中IL-2的含量。结果表明,A2、A3、A2+A3、A1+A2+A3能显著提高血清中IL-2含量。黄芪多糖免疫功能的有效组分为A2、A3,尤以A2作用显著。     

剪股颖属植物遗传分化及系统关系的分子标记研究

剪股颖属植物形态变异大、倍性复杂、种间易于杂交,导致异名多、分类混乱。本研究采用4种分子标记技术,对包括匍茎剪股颖、巨序剪股颖及红顶草在内的7种剪股颖,共58个个体的遗传分化和系统关系进行研究。根据谱带清晰程度及多态性,筛选出9个RAPD引物、2个SSR引物、5个ISSR引物及1个SCAR标记,并建立了最适的PCR反应条件。采用最大简约法和距离法对7种剪股颖植物进行系统树分析,并用靴带检验法计算内部分支的支持率。启发式搜索得到的简约树和由UPGMA方法得到的表征树近乎相同,其中,匍茎剪股颖和红顶草构成单系类群并得到100%的置信支持。红顶草显示了匍茎剪股颖特异性特征谱带,但同时具有不同于匍茎剪股颖的遗传分化,支持将红顶草视为匍茎剪股颖一个变种的观点。对匍茎剪股颖与巨序剪股颖间的遗传分化进行了探讨。     

Polymorphism of the beta-casein gene and its associations with breeding value for production traits of Holstein–Friesian bulls

Beta-casein A1 protein variant might be one of the risk factors in the etiology of human disorders like diabetes and ischemic heart disease. However, from the practical perspective selecting for the A2 allele in dairy cattle requires knowledge of whether the A1/A2 polymorphism is associated with breeding values for production traits. A DNA fragment containing A1/A2 polymorphic region was amplified and genotyped using the PCR-ACRS (Amplification Created Restriction Site) technique in 478 Holstein–Friesian bulls yielding the allele frequencies of 0.35 and 0.65 for the A1 and the A2 variants, respectively. A linear regression model was used for testing the association between the polymorphism and breeding values for production traits and showed that the allele coding the A2 protein variant increases breeding values for milk and protein yield and decreases breeding values for fat percentage in milk. Genotyping A.I. bulls at beta-casein locus and preferring A2 allele may lead to two benefits: increasing breeding value for protein yield and decreasing frequency of A1 protein variant, being considered as risk factor for human health.     

6种除草剂对黄河源区栽培草地中毒杂草的防治

以黄河源区果洛州玛沁县建植14年的垂穗披碱草 (Elymus nutans) 栽培草地为研究对象,针对以甘肃马先蒿 (Pedicularis kansuensis) 和黄帚橐吾 (Ligularia virgaurea) 为主的阔叶型毒杂草,选择6种除草剂及5种复配剂进行防除试验,6种除草剂为单一除草剂,即2, 4-滴丁酯(A)、使它隆(B)、龙拳(C)、阔诺(D)、刹阔(E)、苯磺隆(F) ;5种复配剂为2, 4-滴丁酯与其他除草剂的复配剂,即(A + B)、(A + C)、(A + D)、(A + E)、(A + F)。结果表明,喷施B、F、A + E除草剂不同程度地降低黄帚橐吾的高度、盖度及生物量,且F和A + E对生物量影响显著 (P < 0.05) ;A、E、F、A + B、A + C和A + F 6种除草剂显著降低了甘肃马先蒿的高度、盖度及生物量(P < 0.05);复配除草剂A + F能有效防除群落毒杂草,防除等级达到最高3级,A + E处理下牧草的盖度和生物量显著高于对照 (P < 0.05);喷施除草剂群落物种丰富度指数、Shannon-Wiener指数呈下降趋势,但均匀度指数比对照略高。通过鲜重防效评价了11种处理的防除效果,结果表明,防除黄河源区栽培草地的毒杂草可选择A + E、F、A + F 3种除草剂。     

The comparative pathogenicity of strains of eight serovars and untypable strains of Mannheimia haemolytica in experimental pneumonia of sheep

The experimental induction of pneumonic pasteurellosis in groups of conventionally reared lambs by 8 serovars (A1, A2, A6, A7, A8, A9, T10, and A11) and untypable (UT) strains of Mannheimia haemolytica (Mh) were examined and compared. The groups of lambs were inoculated intratracheally with 1.4 x 10(8) +/- 0.6 x 10(8) (mean +/- SD) colony-forming units of the Mh serovars or UT isolates in the 6-hour log phase of growth. The variables measured as indicators of disease severity were clinical score, percentage lung consolidation and microbiological re-isolation. The clinical parameters for each group were computed daily for 6 days post infection and the lambs which died were necropsied while the remaining lambs were killed on day 7 pi and the extent of lung consolidation was measured. Clinically, the mean scores for the M. haemolytica serovars were A1 (6.1), A2 (18.8), A6 (0.5), A7 (17.4) and A9 (8.5). The mean percent lung lesion scores for M. haemolytica serovars were A1 (12.5), A2 (66.3), A6 (5.0), A7 (51.3), A9 (33.8) and A11 (2.5). The percent mean pneumonic lung lesions recorded for groups inoculated with A2, A7 and A9 were significantly (P < 0.05) higher than the extent of lung lesions in the other groups. A statistically significant correlation was observed between clinical scores and the severity of the lung lesions (r = 0.96, P < 0.01). High titres of M. haemolytica were recovered from lung lesions, with 10 to 100 times the number of organisms inoculated being present in the lung lesions of lambs inoculated with serovars A2 and A7. These data indicate that although M. haemolytica serovars A1, A2, A6, A7, A9 and A11 are important primary lung pathogens of lambs, serovars A2, A7, and A9 are to be regarded as highly virulent strains that have a greater predilection than the other serovars for causing pneumonia in lambs.     

Effects of fluoroquinolones on CYP4501A and 3A in male broilers

The inhibitory effects of fluoroquinolones on the enzyme activity, protein levels and mRNA expression of liver cytochrome P450 (CYP) 1A and 3A were investigated in male broiler chicks. Enrofloxacin (20 mg/kg), sarafloxacin (8 mg/kg) and marbofloxacin (5.5 mg/kg) were administrated in drinking water for 7 consecutive days. A cocktail of the probe drugs caffeine and dapsone was used to determine CYP1A and 3A activity. Western blot analysis and real-time PCR were used to determine the effects on protein levels of CYP1A and 3A, and on CYP1A4, 1A5, 3A37 mRNA levels. Enrofloxacin increased the half-life of elimination for both caffeine and dapsone, and decreased expression of CYP1A and 3A protein. Marbofloxacin decreased the metabolism of caffeine and expression of CYP1A protein. However, no change in mRNA expression was observed for any treatment group. This suggested that high doses of enrofloxacin and marbofloxacin, but not sarafloxacin, inhibit CYP in chick liver raising the possibility of drug-drug interaction when using these compounds.