Alzheimer病患者APP及PS1基因表达量的研究

为了检测Alzheimer病（Alzheimer’s disease,AD）患者外周血中淀粉样前体蛋白（Amyloid Precursor Protein, APP）基因及早老素1（Presenilin 1, PS1）基因的表达情况,进而探讨APP及PS1基因的表达与AD的相关性,采用SYBRGreenⅠ的方法对45例AD患者、25例血管性痴呆（vascular dementia, VD）患者及60名正常对照组样本的mRNA进行绝对定量,检测得到APP基因及PS1基因在对照组中的表达水平分别为0.026±0.005 amol/μg cDNA和0.026±0.004 amol/μg cDNA;在AD患者组中的表达量分别为0.044±0.006 amol/μg cDNA和0.051±0.011 amol/μg cDNA;,在VD患者组中的表达水平分别为0.072±0.013 amol/μg cDNA和0.039±0.005 amol/μg cDNA 。经显著性检验,AD患者组APP基因的表达水平上调,t=2.639, P<0.01;PS1基因的表达水平同样呈上调趋势,t=2.173,P<0.05,差异均具有统计学意义。VD患者组APP基因的表达水平上调,t=3.028,P<0.01;PS1基因的表达水平也同样呈上调趋势,t=2.012,P<0.05,均有显著性差异。因此,APP及PS1基因的表达水平的增高并不一定与AD发生特异性关联,而可能与多种导致痴呆的脑部病变发生关联。     

Effect of polyunsaturated fatty acid-enriched phosphatidylcholine and phosphatidylserine on butyrate-induced growth inhibition, differentiation and apoptosis in Caco-2 cells

Phospholipids are fascinating in terms of important bio-functional compounds. The present work investigated the effect of polyunsaturated phosphatidylcholine (PC) and phosphatidylserine (PS) on butyrate-induced growth inhibition, differentiation and apoptosis using Caco-2 cells. Growth inhibition of Caco-2 cells became apparent 24 h after addition of PC while it took 48 h with PS. Alkaline phosphatase activity of Caco-2 cells increased with combined PC or PS and sodium butyrate (NaBT) at 72 h, indicating that PC and PS enhanced cell differentiation in the presence of NaBT. An increased enrichment factor was also found when cells were treated with combinations of PC or PS and NaBT. These results suggest that marine PC and PS can be considered to be potentially useful colon cancer chemotherapy agents with high bio-availability.     

Relationship between δ13C and photosynthetic parameters and their responses to leaf nitrogen content in six broadleaf tree species

Stable carbon isotope composition (δ¹³C), net photosynthetic rate (P _N), actual quantum yield of photosystem 2 (PS2) electron transport (ΦPS2), nitrogen content (N_c), and photosynthetic nitrogen use efficiency (PNUE) in the leaves of six broadleaf tree species were determined under field environmental conditions. The six tree species were Magnolia liliflora Desr., M. grandiflora Linn., M. denudata Desr., Prunus mume (Sieb.) Sieb. et Zucc. cv. Meiren Men, P. mume (Sieb.) Sieb. et Zucc. f. alphandii (Carr.) Rehd., and P. persica (L.) Batsch. var. rubro-plena. The relationships among δ¹³C, Φ_PS2, P _N, and PNUE, as well as their responses to N_c in the six species were also studied. Both P _N and δ¹³C negatively correlated with N_c, but Φ_PS2 positively correlated with N_c. This indicated that with N_c increase, P _N and δ¹³C decreased, while Φ_PS2 increased. There were weak negative correlations between δ¹³C and PNUE, and strong negative correlations (p<0.01) between Φ_PS2 and PNUE. According to the variance analysis of parameters, there existed significant interspecific differences (p<0.001) of δ¹³C, P _N, Φ_PS2, PNUE, and N_c among the tree seedlings of the six tree species, which suggests that the potential photosynthetic capacities depend on plant species, irradiance, and water use capacity under field conditions.     

Erythropoietin activates caspase-3 and downregulates CAD during erythroid differentiation in TF-1 cells - a protection mechanism against DNA fragmentation

The involvement of caspase-3 and its failure in the induction of DNA fragmentation during erythropoiesis were investigated with TF-1 cells. During erythroid differentiation, caspase-3 activation and cleavage of caspase-3 substrates such as ICAD (inhibitor of caspase-activated DNase) were detected without concomitant phosphatidyl-serine (PS) externalization and DNA fragmentation. These observations are in contrast to our understanding that DNA is degraded by CAD (caspase-activated DNase) when ICAD is cleaved by caspase-3. Our study demonstrates that CAD is downregulated at the mRNA and protein level during the erythroid differentiation in TF-1 cells. This provides a mechanism for the first time how cells avoid DNA fragmentation with activated caspase-3.     

Identification of novel phospholipid binding proteins in Saccharomyces cerevisiae

A proteomics approach was used to search for novel phospholipid binding proteins in Saccharomyces cerevisiae. Phospholipids were immobilized on a solid support and the lipids were probed with soluble yeast protein extracts. From this, the phosphatidic acid binding proteins were eluted and identified by mass spectrometry. Thirteen proteins were identified and 11 of these were previously unknown lipid binding proteins. The protein-lipid interactions identified would not have been predicted using bioinformatics approaches as none possessed a known lipid binding motif. A subset of the identified proteins was purified to homogeneity and determined to directly bind phospholipids immobilized on a solid support or organized into liposomes. This simple approach could be systematically applied to perform an exhaustive screen for soluble lipid binding proteins in S. cerevisiae or other organisms.     

Role of the leader sequence in tobacco pectin methylesterase secretion

We report that unprocessed tobacco pectin methylesterase (PME) contains N-terminal pro-sequence including the transmembrane (TM) domain and spacer segment preceding the mature PME. The mature portion of PME was replaced by green fluorescent protein (GFP) gene and various deletion mutants of pro-sequence fused to GFP were cloned into binary vectors and agroinjected in Nicotiana benthamiana leaves. The PME pro-sequence delivered GFP to the cell wall (CW). We showed that a transient binding of PME TM domain to endoplasmic reticulum membranes occurs upon its transport to CW. The CW targeting was abolished by various deletions in the TM domain, i.e., anchor domain was essential for secretion of GFP to CW. By contrast, even entire deletion of the spacer segment had no influence on GFP targeting.     

Adaptations in the lipid metabolism of the protozoan parasite Trypanosoma brucei

Trypanosomes are unicellular parasites and like all decent parasites, they try to obtain from the host as much material as possible, including lipids. However, the needs of a parasite are not always the same as those of the host, and therefore, mostly, some biosynthetic work still has to be done by the parasite itself. Very often at least modifications of the lipid components that are acquired from the host have to be made. Furthermore, next to the lipids Trypanosoma brucei indeed obtains from the host, some other lipid components have to be synthesized de novo. Especially the processes where the metabolism of T. brucei differs from that of the host, will be discussed, as at least some of them are excellent targets for the development of urgently needed new chemotherapeutics.     

A polymorphism in New Zealand inbred mouse strains that inactivates phosphatidylcholine transfer protein

New Zealand obese (NZO/HlLt) male mice develop polygenic diabetes and altered phosphatidylcholine metabolism. The gene encoding phosphatidylcholine transfer protein (PC-TP) is sited within the support interval for Nidd3, a recessive NZO-derived locus on Chromosome 11 identified by prior segregation analysis between NZO/HlLt and NON/Lt. Sequence analysis revealed that the NZO-derived PC-TP contained a non-synonymous point mutation that resulted in an Arg120His substitution, which was shared by the related NZB/BlNJ and NZW/LacJ mouse strains. Consistent with the structure-based predictions, functional studies demonstrated that Arg120His PC-TP was inactive, suggesting that this mutation contributes to the deficiencies in phosphatidylcholine metabolism observed in NZO mice.     

An arrayed human genomic library constructed in the PAC shuttle vector pJCPAC-Mam2 for genome-wide association studies and gene therapy

The various iterations of the HapMap Project and many genome-wide association studies (GWAS) have identified hundreds of potential genes involved in monogenic and multifactorial traits. We constructed an arrayed 115,000-member human genomic library in the PAC shuttle vector pJCPAC-Mam2 that can be propagated in both bacterial and human cells. The library appears to represent a two-fold coverage of the human genome. Transient transfection of a p53-containing PAC clone into p53-null Saos-2 human osteosarcoma cells demonstrated that both p53 mRNA and protein were produced. Additionally, expression of the p53 protein triggered apoptosis in a subset of the Saos-2 cells. This library should serve as a valuable resource to validate potential disease genes identified by GWAS in human cell lines and in animal models. Also, individual library members could potentially be used for gene therapy trials for a variety of recessive disorders.     

Mapping QTLs and candidate genes for iron and zinc concentrations in unpolished rice of Madhukar×Swarna RILs

Identifying QTLs/genes for iron and zinc in rice grains can help in biofortification programs. 168 F(7) RILs derived from Madhukar×Swarna were used to map QTLs for iron and zinc concentrations in unpolished rice grains. Iron ranged from 0.2 to 224ppm and zinc ranged from 0.4 to 104ppm. Genome wide mapping using 101 SSRs and 9 gene specific markers showed 5 QTLs on chromosomes 1, 3, 5, 7 and 12 significantly linked to iron, zinc or both. In all, 14 QTLs were identified for these two traits. QTLs for iron were co-located with QTLs for zinc on chromosomes 7 and 12. In all, ten candidate genes known for iron and zinc homeostasis underlie 12 of the 14 QTLs. Another 6 candidate genes were close to QTLs on chromosomes 3, 5 and 7. Thus the high priority candidate genes for high Fe and Zn in seeds are OsYSL1 and OsMTP1 for iron, OsARD2, OsIRT1, OsNAS1, OsNAS2 for zinc and OsNAS3, OsNRAMP1, Heavy metal ion transport and APRT for both iron and zinc together based on our genetic mapping studies as these genes strictly underlie QTLs. Several elite lines with high Fe, high Zn and both were identified.