Absence of cystathionase in human fetal liver: is cystine essential?

Cystathionase activity is not measurable in the livers of 24 human fetuses and 3 premature infants, and the concentration of cystathionine in the liver is higher than that of the brain. The placenta does not subserve the trans-sulfuration function. Cystine (or cysteine) thus may be an essential amino acid in the immature human.     

Isolation of a cDNA from the virus responsible for enterically transmitted non-A, non-B hepatitis

Major epidemic outbreaks of viral hepatitis in underdeveloped countries result from a type of non-A, non-B hepatitis distinct from the parenterally transmitted form. The viral agent responsible for this form of epidemic, or enterically transmitted non-A, non-B hepatitis (ET-NANBH), has been serially transmitted in cynomolgus macaques (cynos) and has resulted in typical elevation in liver enzymes and the detection of characteristic virus-like particles (VLPs) in both feces and bile. Infectious bile was used for the construction of recombinant complementary DNA libraries. One clone, ET1.1, was exogenous to uninfected human and cyno genomic liver DNA, as well as to genomic DNA from infected cyno liver. ET1.1 did however, hybridize to an approximately 7.6-kilobase RNA species present only in infected cyno liver. The translated nucleic acid sequence of a portion of ET1.1 had a consensus amino acid motif consistent with an RNA-directed RNA polymerase; this enzyme is present in all positive strand RNA viruses. Furthermore, ET1.1 specifically identified similar sequences in complementary DNA prepared from infected human fecal samples collected from five geographically distinct ET-NANBH outbreaks. Therefore, ET1.1 represents a portion of the genome of the principal viral agent, to be named hepatitis E virus, which is responsible for epidemic outbreaks of ET-NANBH.     

Nuclear receptor-dependent bile acid signaling is required for normal liver regeneration

Liver mass depends on one or more unidentified humoral signals that drive regeneration when liver functional capacity is diminished. Bile acids are important liver products, and their levels are tightly regulated. Here, we identify a role for nuclear receptor-dependent bile acid signaling in normal liver regeneration. Elevated bile acid levels accelerate regeneration, and decreased levels inhibit liver regrowth, as does the absence of the primary nuclear bile acid receptor FXR. We propose that FXR activation by increased bile acid flux is a signal of decreased functional capacity of the liver. FXR, and possibly other nuclear receptors, may promote homeostasis not only by regulating expression of appropriate metabolic target genes but also by driving homeotrophic liver growth.     

Presence of nonoxidative ethanol metabolism in human organs commonly damaged by ethanol abuse

Acetaldehyde, the end product of oxidative ethanol metabolism, contributes to alcohol-induced disease in the liver, but cannot account for damage in organs such as the pancreas, heart, or brain, where oxidative metabolism is minimal or absent; nor can it account for the varied patterns of organ damage found in chronic alcoholics. Thus other biochemical mediators may be important in the pathogenesis of alcohol-induced organ damage. Many human organs were found to metabolize ethanol through a recently described nonoxidative pathway to form fatty acid ethyl esters. Organs lacking oxidative alcohol metabolism yet frequently damaged by ethanol abuse have high fatty acid ethyl ester synthetic activities and show substantial transient accumulations of fatty acid ethyl esters. Thus nonoxidative ethanol metabolism in addition to the oxidative pathway may be important in the pathophysiology of ethanol-induced disease in humans.     

人源中心体新基因E9730在大肠杆菌中的表达及其多克隆抗体的制备

E9730是从人胎肝cDNA文库中分离出来的一个中心体新基因,其在mRNA水平上经5’端选择性剪 切为两个剪切体E9730—A和E9730—B,它们分别编码99个和142个氨基酸残基的多肽链。试验成功实现了 E9730-A和E9730—B在大肠杆菌JM109中的高效表达,并制备了相应的多克隆抗体,后经Western blot检测 具有良好的特异性。     

鸡MOB基因的克隆与表达谱分析

摘要：MOB家族是一个庞大的高度保守的蛋白家族,MOB蛋白参与调控细胞周期和细胞形态形成,而且与肿瘤的发生发展可能有关。本研究利用RT-PCR技术克隆获得了鸡的MOB基因全长编码序列并测序验证;利用生物信息学方法对鸡MOB基因的遗传进化性和蛋白功能进行预测分析;并利用半定量RT-PCR技术对雏鸡的MOB基因进行组织表达谱分析。实验结果表明鸡MOB基因核酸和氨基酸序列与人和鼠的同源基因同源性在80%以上;生物信息学对MOB蛋白的功能预测,发现鸡MOB蛋白具有6个跨膜结构域和一个SAM结构域;组织表达谱分析表明鸡MOB基因在各个组织中均有表达,但在脑、肠、脾、肺、骨骼肌和肝脏中表达量较高。含SAM结构域的多种蛋白在胚胎发育、神经形成和白血病等过程中具有重要功能,相信本研究通过对鸡MOB基因的研究能够为人类同源基因的深入研究有所帮助,并希望为人类肿瘤发生机制的深人探讨提供有价值的参考。     

Syrian hamster female protein: analysis of female protein primary structure and gene expression

The concentration in plasma of the female protein (FP) of the golden Syrian hamster is regulated by sex steroids and by mediators of the acute-phase response to tissue injury or inflammation. A complementary DNA (cDNA) clone corresponding to FP was isolated from a hamster liver cDNA library and used to determine the nucleotide sequence and derived amino acid sequence of native FP. The primary sequence of FP is 69 percent identical to human serum amyloid P component and 50 percent identical to human C-reactive protein. Evidence showed that sex-limited and acute-phase control of the FP gene is pretranslational. The FP protein is thus a useful model for investigating dual regulation of expression of a single gene.     

Molecular cloning of two types of GAP complementary DNA from human placenta

The ras p21 GTPase-activating protein (GAP) was purified from human placental tissue. Internal amino acid sequence was obtained from this 120,000-dalton protein and, by means of this sequence, two types of complementary DNA clones were isolated and characterized. One type encoded GAP with a predicted molecular mass of 116,000 daltons and 96% identity with bovine GAP. The messenger RNA of this GAP was detected in human lung, brain, liver, leukocytes, and placenta. The second type appeared to be generated by a differential splicing mechanism and encoded a novel form of GAP with a predicted molecular mass of 100,400 daltons. This protein lacks the hydrophobic amino terminus characteristic of the larger species, but retains GAP activity. The messenger RNA of this type was abundantly expressed in placenta and in several human cell lines, but not in adult tissues.     

Transamination of aminoalkylphosphonic acids with alpha ketoglutarate

Dialyzed homogenates prepared from Escherichia coli, Tetrahymena pyriformis, sea anemone (Anthopleura elegantissima), and mouse liver were tested for ability to transaminate 17 aminoalkylphosphonic acids with alpha-ketoglutarate. 2-Aminoethylphosphonic acid (2-AEP), which occurs naturally in Tetrahymena and anemone, was transaminated by these latter organisms more than any of the substances tested, but not by preparations from liver or E. coli. 3-Aminopropylphosphonic acid was transaminated by all preparations, but much less by Tetrahymena or anemone than was 2-AEP. 2-Amino-3-phosphonopropionic acid was transaminated by all preparations. 2-Amino-4-phosphonobutyric acid was transaminated by three of the preparations, but not by liver. Of the other 13 substances tested, the following gave positive results: DL-1,2-diaminoethylphosphonic acid with E. coli, DL-1,2-diaminoethylphosphonic and aminomethylphosphonic acids with Tetrahymena, DL-1-aminopropylphosphonic acid with anemone, and DL-1-aminoethylphosphonic and DL-1-aminobutylphosphonic acids with liver. The significance of these transaminations is discussed.     

Cloning of human androgen receptor complementary DNA and localization to the X chromosome

The androgen receptor (AR) mediates the actions of male sex steroids. Human AR genomic DNA was cloned from a flow-sorted human X chromosome library by using a consensus nucleotide sequence from the DNA-binding domain of the family of nuclear receptors. The AR gene was localized on the human X chromosome between the centromere and q13. Cloned complementary DNA, selected with an AR-specific oligonucleotide probe, was expressed in monkey kidney (COS) cells and yielded a high-affinity androgen-binding protein with steroid-binding specificity corresponding to that of native AR. A predominant messenger RNA species of 9.6 kilobases was identified in human, rat, and mouse tissues known to contain AR and was undetectable in tissues lacking AR androgen-binding activity, including kidney and liver from androgen-insensitive mice. The deduced amino acid sequence of AR within the DNA-binding domain has highest sequence identity with the progesterone receptor.     

梅花鹿茸胶囊缓解体力疲劳功能研究

目的证明梅花鹿茸胶囊具有缓解体力疲劳功能。方法1.实验动物体重测定法,2.负重游泳法,3.血清尿素测定法,4.肝糖原测定法,5.血乳酸测定法。结果经灌胃给予小白鼠0.15g/kgBW/d、0.30g/kgBW/d和0.60g/kgBW/d梅花鹿茸胶囊内容物（相当于人体推荐摄入量的5倍、10倍、20倍）时,对实验小白鼠的体重未见明显影响。在0.60g/kgBW/d的剂量时,具有延长小白鼠负重游泳时间的作用。梅花鹿茸胶囊内容物在0.30g/kgBW/d和0.60g/kgBW/d的剂量时,可减少运动后小白鼠血清尿素的产生。梅花鹿茸胶囊内容物在0.30g/kgBW/d和0.60g/kgBW/d的剂量时,具有增加小白鼠糖原储备或减少糖原消耗的作用。梅花鹿茸胶囊内容物在0.15g/kgBW/d、0.30g/kgBW/d和0.60g/kgBW/d的剂量时,对运动后小白鼠血乳酸的产生未见明显影响。结论梅花鹿茸胶囊具有缓解体力疲劳功能的作用。     

L-ascorbic acid synthesis in birds: phylogenetic trend

The ability of several species of birds to synthesize L-ascorbic acid is correlated with their phylogeny. In the more primitive species, synthesis of L-ascorbic acid occurs in the kidney. Among the highly evolved passeriform species, kidney and liver can synthesize L-ascorbic acid in some, whereas in others synthesis occurs in the liver. In still others, the capacity for the synthesis of L-ascorbic acid is apparently lost. The pattern of evolution of the ascorbic acid pathway among birds is thus similar to that among mammals.     

Parallel patterns of evolution in the genomes and transcriptomes of humans and chimpanzees

The determination of the chimpanzee genome sequence provides a means to study both structural and functional aspects of the evolution of the human genome. Here we compare humans and chimpanzees with respect to differences in expression levels and protein-coding sequences for genes active in brain, heart, liver, kidney, and testis. We find that the patterns of differences in gene expression and gene sequences are markedly similar. In particular, there is a gradation of selective constraints among the tissues so that the brain shows the least differences between the species whereas liver shows the most. Furthermore, expression levels as well as amino acid sequences of genes active in more tissues have diverged less between the species than have genes active in fewer tissues. In general, these patterns are consistent with a model of neutral evolution with negative selection. However, for X-chromosomal genes expressed in testis, patterns suggestive of positive selection on sequence changes as well as expression changes are seen. Furthermore, although genes expressed in the brain have changed less than have genes expressed in other tissues, in agreement with previous work we find that genes active in brain have accumulated more changes on the human than on the chimpanzee lineage.     

填饲鹅肝脏组织中脂肪酸沉积与FAS基因mRNA的表达丰度

【目的】研究不同填饲期肥肝鹅肝脏组织中不同脂肪酸沉积与脂肪酸合成酶（fatty acid synthetase,FAS）基因mRNA的表达丰度及相关性。【方法】对200只100日龄青农灰鹅进行填饲,从填饲0 d起,每隔6 d屠宰1次,共6次。每次随机选取10只,每只为1个重复,屠宰测定肥肝重、肝中脂肪含量、各种脂肪酸含量和FAS基因mRNA的表达丰度。【结果】①肥肝重随填饲时间的延长而增加,肝中粗脂肪含量（ether extract,EE）随肝重的增加而增加,以填饲18-24 d鹅的肥肝增重最快（504.67 g/6 d）,12-18 d的次之。②肝中饱和脂肪酸（saturated fatty acid,SFA）和单不饱和脂肪酸（monounsaturated fatty acid,MUFA）的含量随着填饲时间的延长而增加,尤其是在12-18和18-24 d这两个阶段的沉积最为明显,而多不饱和脂肪酸（polyunsaturated fatty acid,PUFA）主要是在填饲后期（18-30 d）沉积;在整个填饲过程中,各种脂肪酸的沉积表现为填饲后期显著大于填饲前期（P＜0.05或P＜0.01）;填饲30 d时沉积量较大的脂肪酸为肉豆蔻酸（C14﹕0）、棕榈酸（C16﹕0）、硬脂酸（C18﹕0）、棕榈油酸（C16﹕1）、油酸（C18﹕1）、二十碳烯酸（C20﹕1）和亚油酸（C18﹕2）,每100 g组织中的含量分别为0.6028 、17.72、7.25、2.75、37.42、0.3078和0.43 g。③FAS基因mRNA的表达丰度随肝脏增重和脂肪沉积速度的增加呈现出先增加后迅速降低的趋势,填饲24 d时极显著高于其它时期（P＜0.01）;0-24 d鹅肝脏组织中FAS基因mRNA表达丰度与肥肝重、EE、SFA和MUFA存在极显著正相关（P＜0.01）,而与PUFA存在弱负相关且差异不显著（P＞0.05）,30 d时相关性不显著（P＞0.05）。【结论】①鹅肝中EE、SFA和MUFA的含量随填饲时间和肝重的增加而增加;填饲18-24 d鹅肝脏增重和EE、SFA和MUFA的沉积速度最快;②FAS基因mRNA的表达对肥肝鹅肝脏中脂肪的沉积具有填饲前、中期快速增加,填饲后期下降的调控作用。     

Aldh3a2a敲除对斑马鱼体色的影响

本研究探讨了Aldh3a2a在斑马鱼色素细胞发育中的意义及其对醛代谢和肝脏健康的影响。利用CRISPR/Cas9技术,我们成功敲除斑马鱼体内的aldh3a2a,导致黑色素细胞、黄色素细胞和虹彩细胞合成数量减少。转录组分析揭示了基因表达谱的变化,特别是与钾通道和类固醇生物合成途径相关的基因表达谱。我们的研究结果表明,aldh3a2a和aldh1a2之间存在调控关系,对视黄酸受体信号传导有影响。GO和KEGG富集分析强调了对色素细胞发育以外的细胞过程的更广泛影响。此外,aldh3a2a基因敲除会导致醛积累,导致肝脏异常,类似于Sj?gren-Larsson综合征,本研究揭示了aldh3a2a在斑马鱼中的多方面作用及其与人类遗传疾病的相关性。     

中国肝细胞癌p53基因热点突变R249S的载体构建及其在细胞中的表达

抑癌基因p53突变是人肝细胞癌（Hepatocellular carcinoma，HCC）中常见的现象，中国江苏启东地区的HCC患者中p53基因的第249位密码子突变AGG→AGT/Arg→Ser（R249S）被认为是热点突变。本研究以人肝cDNA为模板，扩增人全长p53基因，将其克隆到pCMV-Myc载体中。利用定点突变引物以pC-MV-p53为模板，构建R249S定点突变体pCMV-R249S，转染p53缺陷型的H1299细胞系。Western Blot检测表明，克隆在pCMV-Myc载体中的p53基因和249位密码子热点突变的p53基因都可以在H1299细胞系中表达。     

黄颡鱼繁殖期主要组织器官中氨基酸和脂肪酸的组成

采用生物化学方法,对黄颡鱼繁殖期的性腺、肝脏、肌肉中的氨基酸和脂肪酸进行了测定.结果表明:氨基酸组成中,雄性亲鱼肌肉中氨基酸总量最高,为42.73 g/100g;其次是精巢,为36.40 g/100g;肝脏中含量最低,为24.01 g/100g.含量较高的氨基酸有Glu,Asp,Lys,Leu,Arg,Ala,Gly,Phe.Glu在所有氨基酸中含量最高,Lys在必需氨基酸中含量最高.雌性亲鱼肌肉中氨基酸总量最高,为41.98 g/100g;其次是卵巢,为25.71 g/100g;肝脏中含量最低,为18.74 g/100g.脂肪酸:雌雄亲鱼均为MUFA所占比例最高,其次是SFA,PUFA比例最小.脂肪酸比例较高的雄鱼与雌鱼相似,分别是C18:1n-9(油酸),C16:0(软脂酸),C18:2n-6(亚油酸),C16:1n-7(棕榈酸),C18:0(硬脂酸).研究揭示了黄颡鱼繁殖期性腺、肝脏、肌肉在物质组成上的特点.     

延边黄牛转铁蛋白受体2基因克隆与序列分析

【目的】克隆延边黄牛转铁蛋白受体2（transferrin receptor 2,TFR2）基因,并分析该基因在不同组织器官中的表达分布。【方法】通过提取肝脏组织总RNA,采用RT-PCR及RACE方法克隆延边黄牛TfR2基因,采用生物信息学软件进行序列分析,用半定量PCR（SqRT-PCR）方法分析TfR2基因在不同组织器官中的表达分布规律。【结果】①成功克隆出了延边黄牛TfR2基因完整ORF区及3′UTR区（GenBank登录号：GU553087）,该基因全长2 901 bp,ORF区2 412 bp,编码803个氨基酸;②延边黄牛TfR2基因核苷酸序列与其它物种的同源性在80%以上,不同物种间TfR2基因氨基酸序列,尤其是影响其功能的关键氨基酸位点高度保守;③各物种TfR2蛋白功能结构域同样高度保守;④牛TfR2基因主要在肝脏中表达,其它组织,如脾脏、心脏、肾脏和肠道（十二指肠）中也存在少量表达。【结论】不同物种间TfR2基因具有高度同源性,功能结构域及组织分布规律具有高度保守性。     

外源性环核苷酸对枯草期同羊肉脂品质的影响

用标准及自制的环核苷酸组合剂分别处理青年羯同羊各１２只，观测其胴体组成、肉质、肝脏、脂肪和血液等性状指标。结果表明，环核苷酸能显著改善同羊的胴体组成，肉质干物质及粗蛋白含量显著增加，脂肪含量显著下降；体脂中对人体必需的不饱和脂肪酸含量及尾脂中饱和脂肪酸含量均显著增加，体、尾脂中癸酸含量显著下降。