SIRT1过表达慢病毒稳转细胞系的构建及其对脂肪合成的影响

目的：通过油红O及BODIPY染色选取脂肪代谢理想的细胞模型,并运用高内涵仪器检测SIRT1高表达细胞系中脂肪的变化。方法：在L-02、HepG2、Huh7细胞中进行油红O和BODIPY染色,观察在不同油酸浓度下脂滴的生成情况;将构建成功的SIRT1过表达慢病毒载体GV166．SIRT1及空载体病毒GV166．Control感染L．02细胞,qPCR及Western．Blot检测感染细胞中SIRT1的表达水平;高内涵系统检测L-02SIRT1和L-02control细胞系产生的脂滴荧光强度,以确定油酸诱导的最佳浓度及最佳刺激时间。结果：通过油红O及BODIPY染色发现L-02细胞更适宜作为脂肪代谢的细胞模型;成功构建SIRT1过表达慢病毒载体GV166-SIRT1及空载体病毒GV166．Control,qPCR及Western．Blot检测显示转染病毒后SIRT1在细胞中的表达水平明显升高;在油酸刺激浓度0．4mM、诱导时间12h时,L-02SIRT1细胞中脂滴的荧光强度明显较L．02control为低。且这种差异达到最大化。结论：成功建立SIRT1过表达稳定转染细胞系,并证明高表达SIRT1能够抑制脂肪合成。     

Analysis of genetic diversity and phylogenetic relationships among red jungle fowls and Chinese domestic fowls

Genetic diversity and phylogenetic relationships among 568 individuals of two red jungle fowl subspe- cies (Gallus gallus spadiceus in China and Gallus gallus gallus in Thailand) and 14 Chinese domestic chicken breeds were evaluated with 29 microstaellite loci, the genetic variability within population and genetic differentiation among population were estimated, and then genetic diversity and phylogenetic relationships were analyzed among red jungle fowls and Chinese domestic fowls. A total of 286 alleles were detected in 16 population with 29 microsatellite markers and the average number of the alleles observed in 29 microsatellite loci was 9.86±6.36. The overall expected heterozygosity of all population was 0.6708±0.0251, and the number of population deviated from Hardy-Weinberg equilibrium per locus ranged from 0 to 7. In the whole population, the average of genetic differentiation among population, measured as FST value, was 16.7% (P<0.001), and all loci contributed significantly (P<0.001) to this differentiation. It can also be seen that the deficit of heterozygotes was very high (0.015) (P<0.01). Reynolds' distance values varied between 0.036 (Xiaoshan chicken-Luyuan chicken pair) and 0.330 (G. gallus gallus-Gushi chicken pair). The Nm value ranged from 0.533 (between G. gallus gallus and Gushi chicken) to 5.833 (between Xiaoshan chicken and Luyuan chicken). An unrooted consensus tree was constructed using the neighbour-joining method and the Reynolds' genetic distance. The heavy-body sized chicken breeds, Luyuan chicken, Xiaoshan chicken, Beijing Fatty chicken, Henan Game chicken, Huainan Partridge and Langshan chicken formed one branch, and it had a close genetic relationship between Xiaoshan chicken-Luyuan chicken pair and Chahua chicken-Tibetan chicken pair. Chahua chicken and Tibetan chicken had closer genetic relationship with these two subspecies of red jungle fowl than other domestic chicken breeds. G. gallus spadiceus showed closer phylogenetic relationship with Chinese domestic chicken breeds than G. gallus gallus. All 29 microstaellite loci in this study showed high levels of polymorphism and significant genetic differentiation was observed among two subspecies of red jungle fowl and 14 Chinese domestic chicken breeds. The evolutional dendrogram is as follows: evolutional breeds→primitive breeds (Chahua chicken and Tibetan)→red jungle fowl in China (G. gallus spadiceus)→red jungle fowl in Thailand (G. gallus gallus). The results supported the theory that the domestic fowls might originate from different subspecies of red jungle fowl and Chinese domestic fowls had independent origin.     

Characterization of the Porphyridium cruentum Chl a-binding LHC by in vitro reconstitution: LHCaR1 binds 8 Chl a molecules and proportionately more carotenoids than CAB proteins

The Porphyridium cruentum light harvesting complex (LHC) binds Chl a, zeaxanthin and -carotene and comprises at least 6 polypeptides of a multigene family. We describe the first in vitro reconstitution of a red algal light-harvesting protein (LHCaR1) with Chl a/carotenoid extracts from P. cruentum. The reconstituted pigment complex (rLHCaR1) is spectrally similar to the native LHC I, with an absorption maximum at 670 nm, a 77 K fluorescence emission peak at 677 nm (ex. 440 nm), and similar circular dichroism spectra. Molar ratios of 4.0 zeaxanthin, 0.3 -carotene and 8.2 Chl a per polypeptide for rLHCaR1 are similar to those of the native LHC I complex (3.1 zeaxanthin, 0.5 -carotene, 8.5 Chl a). The binding of 8 Chl a molecules per apoprotein is consistent with 8 putative Chl-binding sites in the predicted transmembrane helices of LHCaR1. Two of the putative Chl a binding sites (helix 2) in LHCaR1 were assigned to Chl b in Chl a/b-binding (CAB) LHC II [Kühlbrandt et al. (1994) Nature 367: 614–21]. This suggests either that discrimination for binding of Chl a or Chl b is not very specific at these sites or that specificity of binding sites evolved separately in CAB proteins. LHCaR1 can be reconstituted with varying ratios of carotenoids, consistent with our previous observation that the carotenoid to Chl ratio is substantially higher in P. cruentum grown under high irradiance. Also notable is that zeaxanthin does not act as an accessory light-harvesting pigment, even though it is highly likely that it occupies the position assigned to lutein in the CAB LHCs.This revised version was published online in October 2005 with corrections to the Cover Date.     

江西铅山红芽芋脱毒苗球茎愈伤组织诱导及其再生体系的建立

以江西铅山红芽芋脱毒苗为试材,研究不同因素对红芽芋脱毒苗球茎愈伤组织诱导及其再生体系的影响,以期对红芽芋脱毒苗的再生体系进行优化。结果表明,红芽芋脱毒苗球茎愈伤组织诱导的最佳培养基是MS+TDZ 2 mg·L^-1+2,4-D 1 mg·L^-1。红芽芋脱毒苗球茎愈伤组织分化的最佳培养基是MS+TDZ 2 mg·L^-1+NAA 1 mg·L^-1。红芽芋脱毒苗不定芽生根的最佳培养基是1/2MS+NAA 0.5 mg·L^-1+PP₃₃₃ 0.5 mg·L^-1。红芽芋再生苗最好的移栽基质为发酵后的腐锯木屑。红芽芋脱毒苗球茎愈伤组织再生苗移栽时最佳的PP₃₃₃浓度为20～50 mg·L^-1。本试验成功建立了红芽芋脱毒苗球茎愈伤组织的再生体系,为红芽芋脱毒苗转基因的研究和种质创新奠定了基础。     

Photoinhibition of white clover seed germination at low water potential

Photosensitivity of germination of white clover ( Trifolium repens L. cv. Podkowa) seeds was studied under water deficit (low water potential) conditions at 25°C. The seeds showed negative photoblastism, which was most pronounced at -0.03 MPa polyethylene glycol solution. Inhibition was observed at two different wavelength bands with maxima at 660 nm (R) and around 730 nm (FR). Red light acted identically to white light (maximum inhibition ca 50%). The effect of far-red illumination was less inhibitory (20–30%). The photoresponse required long illuminations (3 h exposures); saturation level was at 0.1 W m⁻², independently of the light quality. White clover seed germination showed no reversibility of the effects of R and FR light. Prolonged illumination with R and FR increased the inhibition, and intermittent illumination had a higher effect than a continuous one. It was concluded that the photoinhibition of germination of seeds of Trifolium repens involves a reaction dependent on the rate of phytochrome interconversion, a property that is characteristic for the high irradiance reaction.     

Effects of Korean Red Ginseng extract on hepatic lipid accumulation in HepG2 cells

In this study, we investigated the effects of Korean red ginseng water extract (KRGE) on hepatic lipid accumulation in HepG2 cells. KRGE decreased hepatic triglyceride and cholesterol levels. Further, KRGE suppressed expression of fatty acid synthase (FAS) and 3-hydroxy-3-methyl glutaryl coenzyme A (HMG-CoA) reductase. These results suggest that KRGE may reduce hepatic lipid accumulation by inhibition of FAS and HMG-CoA reductase expression in HepG2 cells.     

中国南部沿海增养殖业的调查与展望 ——来自广东饶平柘林湾和南澳深澳湾“海上渔村“的思考

针对沿海增养殖业对海洋生态的影响,对中国南部沿海2个增养殖业发达区——南澳岛深澳湾"海上渔村"和赤潮灾区饶平柘林湾进行了实地考察和取样分析.考察发现,目前中国南部沿海增养殖业普遍存在技术落后、缺乏科学技术指导和统一管理等问题, 对海洋生态造成了极大的危害;通过海水水质测定实验,发现这2个养殖区水样的平均含氮量较高(2.154 mg/L),氮磷质量比较高(22),处于富营养化状态;浮游植物平均密度较高(1 908.6个/mL),个体含叶绿素a量较低(1.74×10-6 mg/个),出现生态衰退现象.     

Allozyme differentiation within and between red drum (Sciaenops ocellatus) from the Gulf of Mexico and Atlantic Ocean

Nine polymorphic nuclear-gene (allozyme) loci were surveyed among 491 red drum ( Sciaenops ocellatus ) sampled in 1988 and 1989 from nearshore localities in the northern Gulf of Mexico (Gulf) and the Atlantic coast of the southeastern United States (Atlantic). Data were combined with those from a previous study to generate a data set of 762 individuals representing 11 sample localities in the Gulf and 175 individuals representing five sample localities in the Atlantic. The combined data set included individuals from the 1986 and 1987 year classes and permitted rigorous testing of both temporal and spatial genetic heterogeneity. Average heterozygosity-per-locus values (estimated using 33 assumed monomorphic loci) were 0·048 (Gulf red drum) and 0·046 (Atlantic red drum). Tests of heterogeneity in allele frequencies between year classes at individual localities and across regions (Gulf and Atlantic) were non-significant. Tests of spatial (geographic) heterogeneity indicated that red drum are weakly subdivided: genetically-differentiated subpopulations occur in the northern Gulf and along the south-eastern Atlantic coast. Genetic data were consistent with the hypothesis that red drum within the Gulf and along the Atlantic coast comprise singie subpopulalions. Genetic differences between Gulf and Atlantic red drum seem likeiy to stem from historical or recent interactions between dispersal and impediments to gene flow.     

How erythrocyte volume is regulated, or what mathematical models can and cannot do for biology

Modern concepts of the red blood cell (RBC) volume regulation are considered. It is shown that the system of ion pumps and channels in the cell membrane ensures the physiological value of volume with a precision of about 10% even at 5- to 7-fold variations of passive membrane permeability for ions. Particular attention is paid to mathematical models for evaluation of the role of different molecular mechanisms in the RBC volume control. It is shown that many questions, for example, ‘why the Na⁺,K⁺-ATPase pumps the ions in opposite directions’ or ‘what is the physiological role of Ca²⁺-activated K⁺-channels’, cannot be answered without adequate mathematical models of such complex control systems as cell volume control.