高血压患者空腹血游离脂肪酸组成与腰臀比、胰岛素活性的关系

目的观察原发性高血压患者空腹血游离脂肪酸(freefattyacid,FFA)组成与向心性肥胖、胰岛素抗性、年龄、血清锌和饮食的关系及性别差异。方法采用社区人群整群抽样、病例对照方法,筛选出高血压患者109例,其中肥胖患者70例,非肥胖患者39例,并以饮食习惯相近的123例血压正常个体(其中肥胖个体66例,非肥胖者57人)为对照,用高效液相色谱直接衍生化法检测空腹血清各种FFA浓度,放免法测定胰岛素水平。结果(1)多元逐步线性回归和分层分析显示空腹血清FFA组成(包括n3PUFA水平、P/S比值、C20:5/C18:3、C22:6/C20:5及C20:4/C18:2比值)存在年龄和性别差异,即小于45岁年龄组高血压患者的FFA改变显著性大与超过46岁年龄组,男性高血压患者的FFA改变显著性大于女性。(2)分层、相关及回归分析显示空腹血清FFA组成与腰臀围比值、胰岛素敏感指数、血清锌水平、饮食习惯相关(均P<0.05)。(3)原发性高血压患者血清锌水平略低于非高血压个体(14.9±5.0vs非高血压组16.8±6.4)μmol/L,P=0.45。结论原发性高血压患者空腹血游离脂肪酸组成改变与向心性肥胖、胰岛素抵抗、锌缺乏、年龄和饮食有关,并存在性别差异。

Analysis of Hereditary Stability and Disease Susceptivity of sFat-1 Transgenic Pigs

[Objective] This study aimed to investigate the hereditary stability of sFat-1 transgenic pigs and the differences in disease susceptivity between sFat-1 transgenic pigs and non-transgenic pigs. [Method] The integration of sFat-1 gene in pigs was detected by PCR; the infection of transgenic pig to pseudorabies, leptospirosis, swine dysentery, brucellosis, Mycobacterium tuberculosis, rotavirus and mycoplasma hyopneumoniae was detected by using ELISA and PCR. [Result] The positive ratio of F3 generation sFat-1 transgenic pigs was 18.5%; the susceptivity of positive sFat- 1 transgenic and negative pigs to seven infectious diseases showed no significant difference. [Conclusion] Exogenous gene in sFat-1 transgenic pigs can not be stably inherited. The overall physical condition of positive transgenic and negative pigs was similar.

Cloning and molecular characterization of△^(12)-fatty acid desaturase gene from Mortierella isabellina

AIM： To clone △^12 -fatty acid desaturase gene of Mortierella isabellina and to functionally characterize this gene in vitro and in vivo.METHODS： Reverse transcriptional polymerase chain reaction （RT-PCR） was used to clone the open reading frame of △^12-fatty acid desaturase gene （D12D） of Mortierella isabellina. Plasmids pEMICL12 and pYMICL12 were constructed with it. pEMICL12 was transformed into Escherichia coli（E.coli） strain BL21 using CaCl2 method for expression after induction with IPTG. pTMICL12 was transformed into Saccharomyces cerevisiae strain IN- VSc1 using lithium acetate method for expression under the induction of galactose. Northern blotting method was used to investigate the effect of temperature on the transcriptional level of this gene in S.cerevisiae strain IN- VSc1.RESULTS： Recombinant plasmids pEMICL12 and pTMICL12 were successfully constructed and transformed into E. coli and S.cerevisiae separately with appropriate method. After induction with IPTG and galactose, it was found that expression of △^12-fatty acid desaturase genes in E.coli and S. cerevisiae under appropriate conditions led to the production of active △^12-fatty acid desaturase, which could convert 17.876% and 17.604% of oleic acid respectively to linoleic acid by GC-MS detection in vitro and in vivo.CONCLUSION： Cloning and expression of M.isabellina D12D gene in E.coli and S.cerevisiae is successfully completed.

Cloning, Expression of Crocus sativus Phytoene Desaturase Gene and Preparation of Antiserum against It

A 2 149 bp full length phytoene desaturase (PDS) cDNA was first cloned from saffron (Crocus sativus L.) stigma using RT-PCR technique and a rapid amplification of cDNA end (RACE) strategy. The cDNA has an open reading frame of 1 697 bp, which encodes a polypeptide of 565 amino acids. The coding region of the cDNA was inserted into a prokaryotic expression vector pET-21a(+) and over-expressed inE. coli BL21 (DE3). The fusion proteins were found largely in an insoluble inclusion bodies. The purified fusion protein was used to immunize rabbits to obtain polyclonal antiserum with titer of 1×105. Western blot analysis by using this particular antiserum showed that the higher expression level of PDS in mature stigma than in leaves and stamen, and the higher expression level of PDS in mature stigma than in young stigma. Key words saffron - carotenoids - phytoene desaturase - gene expression - antiserum - Western blot CLC number Q 781 - Q 786 Foundation item: Supported by the Doctoral Foundation of the Ministry of Education, P. R. China and the Young Science Foundation of Sichuan University (Grant 0020405505012)Biography: Bai Jie (1968-), female, Ph. D candidate, research direction: plant developmental biology and reproductive engineering.

Transcriptomic analysis of Synechocystis sp. PCC6803 under low-temperature stress

In this study, cDNA microarrays were developed from 3569 mRNA reads to analyze the expression profiles of the transcriptomes of Synechocystis sp. PCC6803 under low temperature （LT） stress. Among the genes on the cDNA microarrays, 899 LT-affected genes exhibited a 1.5-fold （or greater） difference in expression compared with the genes from normal unstressed Synechocystis sp. PCC6803. Of the differentially expressed genes, 353 were up-regulated and 246 were down-regulated. The results showed that genes involved in photosynthesis were activated at LT （10℃）, including genes for photosystem I, photosystem II, photosynthetic electron transport, and cytochrome b6/f complex. Moreover, desg, one of four genes that encode the fatty acid desaturases, was also induced by LT. However, the LT conditions to some degree enhanced the transcription of some genes. In addition, LT （10℃） may reduce cellular motility by regulating the transcription of spkA （sll1575）, a serine/threonine protein kinase. The results reported in this study may contribute to a better understanding of the responses of the Synechocystis cell to LT, including pathways involved in photosynthesis and repair.

Cloning and Molecular Identification of A Fatty Acid Desaturase 2 Gene in a and C Genome of Brassica Species

The fatty acid desaturase 2(fad2) gene was proven to be a major locus for high oleic acid(C18:1).Brassica napus is an amphidiploid species originating from a spontaneous hybridization of Brassica rapa and Brassica oleracea.B.napus contains multiple copies in genome for most of the genes,including fad2 genes.The research cloned nine fad2 genes from 3 varieties of B.rapa and 3 varieties of B.oleracea,respectively.Alignment of the nine fad2 sequences from B.rapa and B.oleracea detected 6 single nucleotide polymorphic sites,which resulted in 6 amino-acid substitutions.The nucleotide substitutions at position 743 bp in the fad2-A gene and position 947 bp in the fad2-C gene were used as 3' end of allele-specific primers.In use of the allele-specific primers to amplify fad2 gene,we could identify if the fad2 gene originated from A genome or C genome.Besides,the research found that fad2 genes in C genome are more conserved in evolutionary process than those in A genome.The fad2 expression data reported in this study revealed that fad2-A from B.rapa was not only expressed in siliques same as fad2-C from B.oleracea,but also expressed in a high level in stems.Not even the less,fad2 gene from B.napus was expressed higher in roots and flowers.All these results provided evidences that fad2,though it was expressed differently in B.rapa and B.oleracea,but it was regulated by the same approach in B.napus.