Effect of Indoor Concentrate Feeding vs. Outdoor Grazing on the Expression of Genes Involved in Muscle Growth and Nutrient Content in Japanese Black Steer Muscle

The objective of the present study was to investigate the effect of outdoor grazing on the expression of genes involved in muscle growth and the nutrient contents of skeletal muscle in steers. Ten Japanese Black steers were divided into two groups: grazing (GR) and concentrate (CT) groups. Crude protein, extractable lipid, moisture, fatty acid, cooking loss and Warner Bratzler shear force in muscle tissue were analyzed. The gene expression of myosin heavy chain (MyHC) isoform (2a, 2x and slow), myostatin, follistatin, peroxisome proliferator-activated receptor γ2 (PPARγ2), CCAAT/enhancer binding protein α (C/EBPα), heat shock protein (HSP) 27 and HSP40 in skeletal muscles was evaluated at the end of fattening. Decreases in MyHC-2a and MyHC-2x (fast-twitch fiber type) expression in the longissimus lumborum (LL) muscle were detected in the GR group compared with the CT group;in contrast, an increase in MyHC-slow (slow-twitch fiber type) expression was shown in the GR group. These results suggest that grazing initiated muscle fiber type conversion to slow-twitch from fast-twitch. A decrease in extractable lipid content was observed in the GR group in the LL and semitendinosus (ST) muscles. Crude protein content in the LL muscle in the GR group was higher than in the CT group. MyHC expression in LL muscle in the GR group was greater than in the CT group. A decrease in myostatin and PPARγ2 gene expression was detected in the GR group compared with the CT group in both muscles. Expression of C/EBPα in LL muscle in the GR group was lower than in the CT group. These results suggest that grazing steers at the end of fattening may lead to an increase in protein content and a decrease in fat accumulation in LL and/or ST muscles by regulation of myostatin, MyHC, PPARγ2 and C/EBPα gene expression.

Escherichia coli Expression of Lactate Dehydrogenase C Gene of Black-Lipped Pika in Western Sichuan Plateau

[ Objective] To express lactate dehydrogenase C （LDH-C） gene in prokaryotic system and then purify the recombinant protein. E Meth- od] The LDI-I-C gene was amplified from black-lipped pika（ Ochotona curzoniae） by RT-PCR and ligated to the expression vector pET-32a. The E. coli BL21 （DE3） carrying the LDI-I-C gene was obtained and induced by IPTG （isopropy-beta-D-thiogalactoside）. The expression products were analyzed by SDS-PAGE and purified by affinity chromatography. L Result] An approximately 1.0 kbp band appeared in the RT-PCR products as the- oretically expected. As evidenced by PCR and double enzyme digestion, an approximately 1.0 kbp fragment appeared, which indicated successful construction of expression vector. As analyzed by SDS-PAGE, a fusion protein with molecular weight slightly larger than 45 kDa was expressed in form of inclusion body. And very pure fusion protein was obtained by nickel ions-charged affinity chromatography. [ Coaclusion] The LDH-C gene of black-lipped pika has been cloned and expressed in E. coil

黑莓品种‘Black Butte’的组织培养

以黑莓品种‘Black Butte’的带芽茎段为外植体进行组织培养研究。结果表明,基本培养基的成分、激素种类和浓度对‘Black Butte’不定芽增殖和生长均有显著影响。综合分析认为,较适基本培养基是添加0.05mmol/L铁盐的MS培养基,较适增殖培养基为MS+0.05 mmol/L铁盐+0.5 mg/L 6-BA+0.3 mg/L IBA,较适生根培养基为1/2 MS+0.05 mmol/L铁盐+0.01 mg/L NAA。

音乐录影带中情感表达的体认多模态隐喻建构——以Black or White音乐录影带为例

自20世纪90年代起,以Forceville为代表的一批国外学者开始着眼于多模态隐喻的研究,探讨图像、声音、语言等不同模态表征符号中的隐喻构建,研究范围涉及广告、漫画、手势、音乐和电影等,但选取MV为研究对象的却很少,尤其是从体认语言学角度研究的几乎没有。有鉴于此,文章选取美国歌手迈克尔·杰克逊以“反种族歧视”为主题的Black or White MV为语料,基于体认多模态隐喻理论,分析其中有关“骄傲”和“愤怒”情感的多模态隐喻建构。分析发现,在该MV中,“骄傲”和“愤怒”情感都涉及非语言听觉和视觉符号的隐喻建构。“骄傲”分别涉及“骄傲是高音”和“骄傲是向上的舞蹈动作”隐喻概念;“愤怒”分别涉及“愤怒是高音”和“愤怒是火”隐喻概念。这些非语言符号隐喻概念都是基于人们的身体体验和认知加工建构起来的,使信息接受主体的情感识解更直接、更快,甚至更强。

Molecular cloning and characterization of the endothelin 3 gene in black bone sheep

Background: Black bone sheep was first discovered in Yunnan province of China in 1970, with unique black pigmentation on the body and internal organs. Endothelin 3(EDN3) has been known as a key gene causing hyperpigmentation in black bone chicken, the Silky fowl.Methods: In this study, EDN3 was employed as a candidate gene for regulating black color pigmentation. First,EDN3 was cloned from sheep to obtain the full-length cDNA by using the rapid amplification of cDNA ends(RACE).Genomic EDN3 was screened and a total of thirty predicted single nucleotide polymorphisms(SNPs) were genotyped for allele and genotype frequency analysis in a case-control study involving two black bone sheep populations. Genomic copy number analysis of EDN3 in sheep was conducted to measure the variation in copy number. EDN3 expression levels were observed among the groups in adult liver, lymph node, and kidney tissues, as well as embryo kidney samples. Also, among the tissues of black bone and non-black bone sheep.Results: The size of the full-length cDNA was 1,578 bp, which included 426 bp of 5′-untranslated region(5′-UTR),an open reading frame(ORF) of 639 bp encoding a protein of 212 amino acids, and a 3′-UTR of 513 bp. Genotype and allele frequencies of all the discovered SNPs were found insignificantly different in black bone and non-black bone sheep(P > 0.05). Genomic copy number analysis of EDN3 in sheep revealed no significant difference between the two sheep groups. No significant variations were found in the adult liver and kidney embryo samples. However,the expression in lymph node and kidney tissue was significantly higher in black bone sheep than that in non-black bone sheep(P < 0.05). Significant variations in the EDN3 expression levels were observed among the tissues of nonblack bone sheep.Conclusions: The findings of the present study indicate that unlike in Silky chickens, EDN3 is not responsible for hyperpigmentation but may play a key functional role in immune and excretory systems of black bone sheep.

A Brief Introduction of Black Humor as A Writing Technique

As a writing technique , black humor has exerted great influence upon modern literature . In this paper , the following issues will be discussed : the differences between traditional humor and black humor , the social reasons which lead to the emergence and development of black humor , the form of expression of black humor .

Triterpene-Enriched Olive Extract as an Immunopotentiator in Black Sea Bream(Acanthopagrus schlegelii)

Recently,the use of natural immunopotentiators,such as plant extracts,is predicted to offer high disease-prevention potential in aquaculture.However,few reports on the immunomodulatory impacts of olive extract(OE)on teleost are available.Therefore,the effects of dietary intake of triterpene-enriched OE on black sea bream(Acanthopagrus schlegelii)were investigated in this study.Our data showed that total blood cell counts,in vivo content of lysozyme,activities of antiprotease and myeloperoxidase,and contents of IL-4 and IL-6 were significantly up-regulated by dietary intake of triterpene-enriched OE.Additionally,the hampered immune response induced by cadmium exposure was significantly mitigated by the administration of OE as were indicated by partially or completely rescued immune-relating parameters.Furthermore,the expressions of immune-related genes encoding NF-κB inhibitor alpha(IkBα),tumor necrosis factorα(TNFα),cyclooxygenase-2(COX-2),and proto-oncogene protein c-fos(FOS)were found to be significantly up-regulated by the dietary intake of OE.In general,the results suggested that the dietary intake of triterpene-enriched OE has immune enhancing effect in black sea bream.Such effect may be realized by 1)increasing the total counts of diverse blood cells;2)activating nonspecific immune biomolecules;and 3)affecting signaling pathways such as IKK and ERK and subsequently inducing IL-4 and IL-6 biosyntheses.

Association of H-FABP gene polymorphisms with intramuscular fat content in Threeyellow chickens and Hetian-black chickens

Background： To explore the relationship between the heart-type fatty acid binding protein（H-FABP） gene and intramuscular fat（IMF）, a polymorphism of the second exon of the H-FABP gene was investigated in 60 Three-yellow chickens（TYCs） and 60 Hetian-black chickens（HTBCs）.Results： The IMF contents of the cardiac, chest and leg muscles in HTBC were increased compared with TYC. Both TYC and HTBC populations exhibited Hardy-Weinberg Equilibrium（HWE） according to the χ2test. Three variations of the two birds were detected, namely, G939 A, G982 A and C1014 T. HTBCs with the TT genotypes exhibit increased IMF content in the chest muscles compared with the TC genotype. Thus, the G982 A site could be considered a genetic marker for selecting increased IMF content in the chest muscles of HTBC. The correlation coefficients revealed that H-FABP m RNA expression was negatively correlated with the IMF content in the cardiac, chest and leg muscles of HTBC and in the cardiac and chest muscles of TYC. The relative m RNA expression of H-FABP was reduced in the cardiac and leg muscles of HTBC compared with TYC, but this difference was not observed at the protein level, as assessed by Western blot analysis.Conclusions： These findings offer essential data that can be useful in the breeding program of HTBC and future research exploring the role of H-FABP in IMF deposition and regulation in chickens.

On the Artistic Expressions of The Color Purple

Black female writer, Alice Walker is always misunderstood as a writer who attaches more attention to the contents and thoughts in her works. But the success of her work The Color Purple fully demonstrates her ability of exquisitely adopting artistic expressions to realize the integration of contents and form. So this paper aims to study three artistic expressions applied in this novel, that is, the epistolary style, symbolism of the real objects and psychological transformation of the characters. Furthermore, it's illustrational to show in this thesis that Alice Walker is not only a writer with deep thoughts, but also an imaginative and creative master of excelling in her writing techniques.

Cloning and Transcriptional Activity of Follicle-Stimulating Hormone Receptor Promoter in the Jintang Black Goat

[ Objective] To clone follicle-stimulating hormone receptor （FSHR） promoter in the Jintang black goat, study its transcriptional activity, and provide a basis for alternative splicing of FSHR gene. [Method] The total DNA were extracted from the womb of Jintang black goat, and one pair of primers were designed for amplification of FSHR promoter fragments, then the sequences and homology were analyzed. The FSHR promoter fragment was inserted into the pcFSHRB1 expression vector to substitute the CMV promoter and construct the pcFSHRB2 expression vector. The pcFSHRB1 and pcFSHRB2 expression vectors were transformed into HEK293 cells, respectively. Then these cells were collected after 24 and 48 h treatment with 2 mlU/ml follicle-stimulating hormone （FSH）, and the cAMP levels were detected. [Result] The FSHR promoter sequence of Jin- tang black goat had 34.2% homology to that of chicken and 41.6% to that of rat, respectively. The transcription initial site of FSHR was at -576 bp and its upstream sequences contained two TATA-boxes, four CAAT-boxes, one E-box and one Wl-box. After treating for 24 and 48 h, the cAMP levels of pcFSHRB2 were respectively 299.581 3 and 125.528 1 pmol/L; and that of pcFSHRB1 were respectively 120.057 1 and 109.940 7 pmoVL. [Conclusion] The FSHR promoter of Jintang black goat is a typical type 2 eukaryotic promoter, and it is also a strong promoter.

赤水乌骨鸡ADSL基因的组织表达特征及其干扰载体构建

【目的】探究赤水乌骨鸡胸肌组织中腺苷琥珀酸裂解酶基因(ADSL)表达特征及其目的基因的干扰载体构建,为深入研究畜禽组织肌肉中肌苷酸(IMP)的表达规律及ADSL基因影响畜禽风味的机制提供理论参考。【方法】以不同月龄(3、4、5和6月龄)的赤水乌骨鸡(公鸡和母鸡各半)为试验材料,运用实时荧光定量PCR检测不同月龄及不同性别赤水乌骨鸡胸肌组织中ADSL基因的表达情况;同时根据GenBank中鸡ADSL基因mRNA序列(NM_205529.2)设计4对短发夹RNA(shRNA)干扰序列(sh1-ADSL、sh2-ADSL、sh3-ADSL和sh4-ADSL)和1对阴性对照序列(sh-NC)与pGPH1/GFP/Neo载体连接后进行测序,将构建成功的ADSL基因干扰载体转染至成肌细胞,检测并筛选出干扰效率最高的干扰载体。【结果】ADSL基因在不同月龄赤水乌骨鸡胸肌组织中的相对表达量排序为5月龄>4月龄>6月龄>3月龄,赤水乌骨母鸡3月龄母鸡的ADSL基因相对表达量极显著高于公鸡(P<0.01,下同),4月龄母鸡ADSL基因的相对表达量显著高于公鸡(P<0.05),6月龄母鸡ADSL基因的相对表达量极显著低于公鸡;ADSL基因质粒的测序结果显示干扰质粒构建成功,质粒转染后干扰组及阴性对照均可见绿色荧光,空白对照不发光,说明各重组质粒成功转染赤水乌骨鸡成肌细胞;ADSL基因沉默结果显示,sh2-ADSL沉默效果最佳,干扰效率为90.30%。【结论】赤水乌骨鸡胸肌中ADSL基因的相对表达量随着月龄增加表现为先上升后下降的变化趋势;3、4和5月龄赤水乌骨母鸡ADSL基因的相对表达量高于公鸡,随着月龄增加ADSL基因的相对表达量增加,公鸡和母鸡间的差异减小。成功分离提取赤水乌骨鸡成肌细胞,并筛选出RNA干扰效果最佳载体sh2-ADSL,为后续研究ADSL基因调控鸡肉肌苷酸及风味的分子机制提供参考。

Brane world black holes in teleparallel theory equivalent to general relativity and their Killing vectors,energy,momentum and angular momentum

The energy--momentum tensor, which is coordinate-independent, is used to calculate energy, momentum and angular momentum of two different tetrad fields. Although, the two tetrad fields reproduce the same space--time their energies are different. Therefore, a regularized expression of the gravitational energy--momentum tensor of the teleparallel equivalent of general relativity （TEGR）, is used to make the energies of the two tetrad fields equal. The definition of the gravitational energy--momentum is used to investigate the energy within the external event horizon. The components of angular momentum associated with these space--times are calculated. In spite of using a static space--time, we get a non-zero component of angular momentum! Therefore, we derive the Killing vectors associated with these space--times using the definition of the Lie derivative of a second rank tensor in the framework of the TEGR to make the picture more clear.

A Mathematical Interpretation of Hawking’s Black Hole Theory by Ricci Flow

In this paper, using Perelman’s no local collapsing theorem and the geometric interpretation of Hamilton’s Harnack expressions along the Ricci flow introduced by R. Hamilton, we present a mathematical interpretation of Hawking’s black hole theory in [1].

大白菜CHS基因鉴定及其在高氮水平下转录表达分析

为了探究高氮水平引起大白菜叶柄黑点症加剧的机制,通过对抗、感叶柄黑点症大白菜品系进行正常氮和高氮水平处理,处理前和处理后不同时间对叶柄取样并进行转录组测序,然后再对大白菜查尔酮合酶(chalcone synthetase,CHS)基因进行鉴定并分析不同的大白菜CHS基因在正常氮水平和高氮水平、抗性品系和感性品系之间的差异表达,结果表明,共鉴定到7个大白菜CHS基因,其中有3个(BrCHS1、BrCHS3及BrCHS4)在高氮水平下表达量比正常氮水平下高,且在高氮水平下感性品系表达量高于抗性品系。因此推测这3个大白菜CHS基因可能与大白菜叶柄黑点症的形成有关。研究结果为揭示大白菜叶柄黑点症发生机制奠定了基础。

拮抗菌株混合发酵液对烟草黑胫病和青枯病的防治作用

黑胫病和青枯病是危害烟草生长的主要土传病害,解淀粉芽孢杆菌(Bacillus amyloliquefaciens)菌株F012和F028对烟草黑胫病和青枯病等多种植物致病菌具有较强拮抗作用。用平板对峙法和盆栽试验研究F012和F028混合发酵液对烟草黑胫病和青枯病的防治作用及机制。结果表明:平板对峙试验中混合发酵液对烟草黑胫病病原菌抑菌率为77.91%,对烟草青枯病病原菌的抑菌圈直径为31.7 mm;盆栽试验中对烟草黑胫病和青枯病防效分别为74.49%和77.27%;混合发酵液能显著上调烟草叶片抗病关键基因NtPAL、NtEDS1、NtPR1和NtPR2的表达,提高苯丙氨酸解氨酶、几丁质酶、β-1,3-葡聚糖酶和多酚氧化酶等保护酶活性。由以上结果可知,F012和F028混合发酵液可以激活烟草防御系统,提高烟草对黑胫病和青枯病的抵抗能力。F012和F028菌株将是研制复合微生物菌剂的理想原料。

宋金时期磁州窑白地黑花瓷缠枝牡丹纹研究

宋金时期磁州窑瓷器装饰以白地黑花装饰最为突出,其创造性地将中国绘画技法以图案装饰形式在瓷器上进行绘制,装饰纹样常有花卉植物纹样等,其中缠枝牡丹纹出现最为频繁。笔者采用资料搜集及对比分析等研究方法,对宋金时期磁州窑白地黑花瓷中缠枝牡丹纹发展脉络进行梳理,归纳分析白地黑花装饰中缠枝牡丹纹的构成组织方式及艺术表现形式,探索其文化底蕴与美学特征,从而发掘其在历史长河中的艺术价值。

浅谈平面设计构成与黑白装饰画表现

黑白装饰画是用单一的颜色在二维平面设计上按照符合审美原则的构成、造型、调性等美术规则进行的绘画创作,亦可视作素描的一种形式。本文通过一系列的实验和研究,试图探讨平面设计构成对黑白装饰画在构图和调性环节进行借用、指导和表现,以期对相同研究领域中的类似情况提供帮助。

贵州黑山羊ADD1基因克隆、生物信息学及组织表达规律研究

为了分析贵州黑山羊ADD1基因编码区序列(coding sequence, CDS)信息及在组织中的表达规律,试验克隆了贵州黑山羊ADD1基因CDS并进行了生物信息学分析,同时分析了该基因在12月龄贵州黑山羊心脏、肝脏、脾脏、肺脏、肾脏、皮下脂肪、背最长肌和腿肌组织中的表达规律,以及在1,3,6,9,12月龄贵州黑山羊皮下脂肪、背最长肌和腿肌组织中的表达时序情况。结果表明:贵州黑山羊ADD1基因CDS大小为2 304 bp,编码767个氨基酸,与山羊ADD1基因序列相似性较高,与禽类的相似性较低;ADD1蛋白中的主要氨基酸为丝氨酸(Ser)、脯氨酸(Pro)、谷氨酸(Glu)和亮氨酸(Leu),含量分别为9.0%、8.7%、8.5%、8.2%,分子质量为84.05 ku,分子式为C_(3682)H_(5840)N_(1036)S_(24),理论等电点为5.74,为不稳定蛋白和亲水蛋白,主要定位在细胞核中,与ADD蛋白家族(ADD2和ADD3)、加帽蛋白(capping actin protein, CAPG)、肌侵蛋白(myotrophin, MTPN)、X盒结合蛋白1(X-box binding protein 1,XBP1)、固醇调节元件结合蛋白1(sterol-regulatory element-binding protein 1,SREBP1)等蛋白存在相互作用,二级结构和三级结构中主要含有α螺旋和无规则卷曲;ADD1基因在12月龄贵州黑山羊心脏、肝脏、脾脏、肺脏、肾脏、皮下脂肪、背最长肌和腿肌组织中均有表达,皮下脂肪中的表达量最高,极显著高于其他组织(P<0.01),其次在心脏、背最长肌、肝脏和腿肌中表达量较高,在脾脏中表达量最低。ADD1基因在1,3月龄贵州黑山羊脂肪、背最长肌和腿肌组织中的表达量均较低。在脂肪组织中6月龄时最高,极显著高于其他月龄(P<0.01);3月龄时最低,极显著低于其他月龄(P<0.01)。在背最长肌中9,12月龄较高,二者差异不显著(P>0.05),但均极显著高于其他月龄(P<0.01);6月龄表达量最低,与1月龄差异不显著(P>0.05),但二者均极显著低于其他月龄(P<0.01)。在腿肌中9月龄时最高,极显著高于其他月龄(P<0.01);12月龄最低,与1月龄差异不显著(P>0.05),但二者均极显著低于其他月龄(P<0.01)。说明ADD1基因在贵州黑山羊各组织中的表达具有广谱性和差异性。

黑色素沉积相关基因在丝羽乌骨鸡和余干乌骨鸡胸肌和腿肌中的表达分析

试验旨在分析黑色素相关基因在丝羽乌骨鸡和余干乌骨鸡中的差异表达。试验测定丝羽乌骨鸡和余干乌骨鸡2~17周龄胸肌和腿肌肉色亮度值,比较黑色素相关基因TYR、TYRP1、PMEL和MITF在两种乌骨鸡胸肌和腿肌的差异表达量。结果显示:2周龄时余干乌骨鸡胸、腿肌和丝羽乌骨鸡胸肌肉色亮度值显著小于其他周龄(P<0.05),余干乌骨鸡6周龄胸、腿肌亮度值显著大于6周龄丝羽乌骨鸡(P<0.05),17周龄时丝羽乌骨鸡腿肌亮度值显著大于余17周龄干乌骨鸡(P<0.05);4个基因在2种乌骨鸡2~17周龄的胸肌和腿肌中相对表达量的变化趋势基本一致,2周龄时表达量较高,随后呈现下降趋势。同时黑色素相关基因表达量存在品种效应,2周龄时TYR和TYRP1基因在丝羽乌骨鸡腿肌中表达量较高,17周龄时MITF基因在余干乌骨鸡腿肌中表达量较高。TYR、TYRP1、PMEL和MITF 4个基因在2种乌骨鸡胸肌和腿肌中表达均呈现显著正相关(P<0.05);4个基因表达与2种乌骨鸡肉色亮度值显著负相关(P<0.05),且对黑色素沉积呈现正调控作用。研究表明,在乌色度选种时可以将TYR、TYRP1和MITF基因作为主效基因进行分子标记辅助选择研究,以培育出适合市场需求的乌色度更高的乌骨鸡新品种。

黑鲷DMRT1基因cDNA的克隆、组织表达谱及在性别逆转前后性腺中的表达

利用RT PCR和 3′ RACE的方法从黑鲷精巢中克隆了DMRT1基因cDNA的部分序列。组织特异性表达分析表明DMRT1基因只在精巢中表达。半定量RT PCR检测显示DMRT1基因在性别逆转前、性别逆转期和性别逆转后的精巢中的表达量无显著差异。在鱼类成体的精巢中 ,DMRT1的表达量可能不会因精巢结构或生理状态的改变而发生改变 。