铜、铁、锌、锰添加水平对蛋青鸡生长性能及养分利用的影响

525只1日龄罗曼褐壳蛋鸡随机分为5处理，每处理7重复，每重复15只鸡。所有处理使用一种基础日粮，统一添加碘和硒。铜、铁、锌、锰四种微量元素添加方案如下，A组不额外添加；B组按50％NRC标准直接添加；C组和D组将日粮铜、铁、锌、锰总量补齐到2倍和3倍NRC水平；E组以基础日粮铁含量与NRC标准比值为基准，将其余微量元素校正到同样倍数。铜、铁、锌、锰添加形式均为硫酸盐。结果表明，1～18周龄生长蛋鸡典型基础日粮中添加铜、铁、锌、锰不影响采食量（P〉0．05）、期末体重和能量表观代谢率（P〉0．05），过量添加降低日增重和饲料转化率（P〈0．05），但提高蛋白质表观代谢率（P〈0．05）。随日粮微量元素水平增加，排泄物铜、锌、锰含量线性增加（P〈0．05），铜、锌、锰表观代谢率呈先增加后降低二次曲线变化（P〈0．05）。西北地区典型蛋青鸡基础日粮中铜、铁，锌、锰已满足需要，无需额外添加。

不同处理下蛋青鸡血浆游离Leu比放射性活度的变化

在绝食、强饲无氮日粮(NFD)、CP20%(豆粕组)、CP20%(棉粕组)4种处理下,给蛋青鸡每公斤体重静脉一次注射1110MBq[3H]Leu,分别于注射后5min,30min,4h,24h,36h,48h翅静脉采血,测定各时间点去蛋白血浆游离Leu浓度和比放射性活度。结果表明,去蛋白血浆中游离Leu浓度绝食组和NFD组显著低于豆粕组和棉粕组(P<0.05);血浆游离Leu的比放射性活度(SRp),以NFD组与绝食组相近,且较高,棉粕组最低,但4种处理间无显著差异(P>0.05)。血浆游离Leu比放射性活度呈显著的指数衰变规律(P<0.05)。

铜、铁、锌、锰添加水平对蛋青鸡生长性能及养分利用的影响

525只1日龄罗曼褐壳蛋鸡随机分为5处理，每处理7重复，每重复15只鸡。所有处理使用一种基础日粮，统一添加碘和硒。铜、铁、锌、锰四种微量元素添加方案如下，A组不额外添加；B组按50％NRC标准直接添加；C组和D组将日粮铜、铁、锌、锰总量补齐到2倍和3倍NRC水平；E组以基础日粮铁含量与NRC标准比值为基准，将其余微量元素校正到同样倍数。铜、铁、锌、锰添加形式均为硫酸盐。结果表明：1～18周龄生长蛋鸡典型基础日粮中添加铜、铁、锌、锰不影响采食量（P〉0．05）、期末体重和能量表观代谢率（P〉0．05），过量添加降低日增重和饲料转化率（P〈0．05），但提高蛋白质表观代谢率（P〈0．05）。

蛋青蛋黄马上见分晓

前些年读大学的人,回家过年时喜欢从所在城市带果脯之类的土特产,不一定好吃,却是游子的一番心意,家中老人都会欢欢喜喜收下。听说现在的大学生更新颖,除了带土特产,还带上恋人,一个鲜亮的大活人,比土特产更能显孝心。

Purification and Identification of Antifreeze Proteins in Ammopiptanthus mongolicus

The conventional protein chromatography technique was adopted to purify the antifreeze proteins (AFPs) from the leaves of Ammopiptanthus mongolicus (Maxim.) Cheng f. Two bands on native PAGE gel showed thermal hysteresis activity, one was band B1, whose thermal hysteresis was 0.46 ℃ at 8 g/L, which showed two bands (67 kD, 21 kD) on SDS_PAGE gel; the other was B3, whose thermal hysteresis was 0.45 ℃ at 10 g/L, and it contained only a single protein (39.8 kD). Both B1 and B3 are not glycoproteins, because neither do they interact with Shiff_reagent, nor show ultraviolet characteristics of a typical glycoprotein.

Plant Anthocyanin Synthesis and Gene Regulation

This paper aims to explain the biochemistry of anthocyanin synthesis based on an overview of plant anthocyanin synthesis genes and environmental factors in the regulation of anthocyanin metabolism. The results show that： ① The metabolism of anthocyanins in plants is affected by the temperature, light, ultraviolet, fertilization status, hormone levels and other factors, which affect the military anthocyanin biosynthetic genes, and then induce or inhibit the synthesis of anthocyanins. ② In the regulation of genes, some of the structural genes of anthocyanin synthesis showed promoting effect, while others showed inhibitory effect. At different environ- mental conditions, the regulation of gene activation and inhibition of the amount of different regulatory genes that anthocyanin accumulation is different, and cause different colors of plant-organs production. ③ In different environmental factors or hor-mones induced to produce the same or different regulation of gene expression changes in regulatory genes, resulting in several different anthocyanins or anthocyanin ratio changes, so that the color of plant organs in different colors.

Construction of Prokaryotic Expression Vectors of EBP1 Gene from Nervilia Fordii (Hance) Schltr.

[Objective] To construct prokaryotic expression vectors encoding gene Erb3binding protein （EBP1）, which plays important roles in regulating plant organ size from Nervilia fordii （Hance） Schltr. [Methods] PCR products of NfEBP1 with particular restriction sites and expression vectors, pET-28 and pET-16b were digested. Ligation, transformation and selection were performed to construct the recombinant plasmids pET-28-NfEBP1 and pET-16-NfEBP1. The recombinant plasmids were transformed into E. coli BL21 using heat -shock transformation. [Results] Recombinant plasmids pET-28-NfEBP1-1188 and pET-16-NfEBP1-1188 were constructed and transformed into expressional host cells, E. coli BL21, and validated by colony PCR, sequencing and double digestion. [Conclusion] Prokaryotic expression vectors of EBP1 gene from N. fordii were successfully constructed, which laid the foundation for characterization of the gene function.

A modified one-step procedure for rapid RNA isolation from insect

A modified guanidinium isothiocyanate method was used to extract total RNA from two forest insect species Clostera anastomosis and Saperda populnea. The integrity of RNA was demonstrated by the methods of gel electrophoresis and cDNA analysis. Typical A260/ A280 absorbance ratio of the total RNA was in range of 1.8 to 2.0. The size of double strand cDNAs obtained by RT-PCR was more than 2 kb, which indicated that intact mRNA was obtained. The fragments of β-actin and chitinase gene from the RNA of C. anastomosis were obtained by RT-PCR, which indicated that the RNA could be used for other molecular operation. By this procedure, RNAs could be extracted and analyzed by electrophoresis from at least 8 samples within 4 hours. These results showed that this method was time- and cost-saving and effective.

从蛋彩画到油画

蛋彩画 油画的前身是蛋彩画(Fresco)。蛋彩画是用颜料同蛋青(或蛋黄和蛋青),再加水、凡立水调和起来后,画在石灰底子墙壁上的一种画,所以蛋彩画又称壁画。蛋彩画在欧洲有很悠久的历史,在古埃及、希腊、罗马和中世纪,这种画法都被广泛地使用。 蛋彩画有充分的表现力,可以画得很细,但它有明显的缺点:一是干得太快,在绘画过程中很难使颜料趁湿进行调和,二是颜色干湿变化较大,干后又很难补笔。

重组PBP2a转肽酶区蛋白核酸适配体的筛选及鉴定

目的筛选和鉴定重组青霉素结合蛋白2a(PBP2a)转肽酶区蛋白的核酸适配体。方法利用指数富集的配体系统进化技术(SELEX),以重组PBP2a转肽酶区蛋白为靶标,从单链DNA文库中筛选能与之特异性结合的核酸适配体。筛选产物克隆测序后,对其进行结构分析和特性鉴定。结果经过11轮筛选,单链DNA文库与靶标蛋白的亲和力趋向稳定,将第8、10轮筛选产物克隆测序,对获得的40个适配体进行分析,一级结构分析显示40个适配体并无共同的保守序列,二级结构预测分析表明,其可分为3个家族。其中13号适配体与重组PBP2a蛋白亲和力最高,并能与之特异性结合。结论利用SELEX技术成功筛选出特异性结合重组PBP2a转肽酶区蛋白的核酸适配体,为探索耐甲氧西林金黄色葡萄球菌感染的诊疗新途径奠定基础。

Research Progress on Heavy Metals Detoxification in Human Body

With the pollution of heavy metals becoming more and more serious, hu- man health suffers from great harms, making the detoxification in human body admit of no delay. This paper summarized the heavy metals used for detoxification in hu- man body in recent years, including metallothionein, polyphenols, dietary fibers from seaweed, GTS, and explored the detoxification mechanism of these heavy metals in human body.

Study on Polymorphisms in the Blood Protein of Tibetan Mastiff

[ Objective] The aim was to study the protein polymorphism in the blood of Tibetan Mastiff, and provide some theoretical basis for resource protection and reasonable development and utilization of Tibetan Mastiff varieties. [ Method] A total of 103 blood samples were taken from four populations of Hequ Tibetan Mastiff, Qinhai Tibetan Mastiff, Tibetan Spaniel and native dogs of Qinghai. Seven blood protein Iocus（Tf, Po, Sα2, Hb, AIb, Pr and Amy）were investigated by using vertical polyacrylamide gel electrophoresis with discontinuous buffer system. Then the genetic variation during different populations was analyzed. [ Result] Genetic variations were observed in Tf, Sα2 and Po in four populations, others were not polymorphic. There were three alleles at the locus of Tf and Po, two alleles at the loci of Sα2. Effective number of alleles and Nei＇s average expected heterozygosity were 1. 532 4 and 0.230 3 relatively, all higher in Tibetan Mastiff than other populations. [ Conclusion] Protein locus in blood of Tibetan Mastiff existed in genetic variation.

Temperature Influences on the Expression of GFP Promoted by the Upstream Sequence of cpcB from Arthrospira platensis

In order to investigate the regulation mechanism of the phycocyanin gene,a series of functional analyses of the upstream sequence of cpcB gene from Arthrospira platensis were conducted in E. coli with green fluorescent protein encoding gene(gfp) as the reporter. Results showed that the gfp gene could express at a high level under the promotion of the upstream sequence,suggesting the existence of some strong promoter elements in it. The expression of GFP was influenced by temperature. Higher temperature led to higher expression level. The bioinformatics analyses followed by mutation analyses on the secondary structure of translation ini-tiation region(TIR) revealed that RNA thermosensor might account for the temperature regulation.

EFFECTS OF PHYCOCYANIN ON EXPRESSION OF CytC mRNA AND CASPASE-3 mRNA AFTER FOCAL CEREBRAL ISCHEMIA/REPERFUSION IN RATS

Objective To study the effects of phycocyanin on the expression of Cytochrome C (CytC)genes and Caspase-3 genes after focal cerebral ischemia/reperfusion in rats. Methods A rat middle cerebral ar-tery occlusion (MCAO)/reperfusion model was produced using the intraluminal filament method. The rats were di-vided into three groups: sham operation group, model control group and phycocyanin group. After MCAO, the neu-robehavioral testing of all rats was made. The infarction area was evaluated with the method of 2,3,7-triphenyltet-razolium chloride (TTC) staining. The expression of CytC mRNA and Caspase-3 mRNA were determined by in situhybridization. Results In the sham operation group and the model control group, there was only a few CytC-positive cells were seen in the normal cerebral tissue. In the model control group, the upregulation of CytC mRNAbegan 6h after ischemia, reached a maximum at 12h (cortex) -24h (striatum) , then subsided gradually, but stillin high level. In the phycocyanin group, CytC-positive cells were also mainly in cortex and striatum, but the numberof the cells was significantly lower than the number of the model control group. The time-phase pattern of CytCmRNA in the phycocyanin group was similar to the pattern of the model control group. In the sham operation groupand the model control group, there was only a few Caspase-3-positive cells were seen in the normal cerebral tissue.In the model control group, the upregulation of Caspase-3 mRNA began 6h after ischemia, reached a maximum at24h and subsided at 48h, but still in high level. In the phycocyanin group, Caspase-3-positive cells were also mainlyin the penumbral area, but the number of the cells were significantly lower than the number of the model controlgroup. The time-phase pattern of Caspase-3 mRNA in the phycocyanin group was similar to the pattern of the modelcontrol group. Conclusion The over-expression of CytC mRNA and Caspase-3 mRNA might play a key role inischemic cerebral injury after MCAO. Phycocyanin could inhibit the over-expression of CytC mRNA and Caspase-3mRNA in the cerebral cortex, and might play an important role in the protection of ischemic neurons.

Non-HFE haemochromatosis

Non-HFE hereditary haemochromatosis （HH） refers to a genetically heterogeneous group of iron overload disorders that are unlinked to mutations in the HFE gene. The four main types of non-HFE HH are caused by mutations in the hemojuvelin, hepcidin, transferrin receptor 2 and ferroportin genes. Juvenile haemochromatosis is an autosomal recessive disorder and can be caused by mutations in either hemojuvelin or hepcidin. Ar~ adult onset form of HH similar to HFE-HH is caused by homozygosity for mutations in transferrin receptor 2. The autosomal dominant iron overload disorder ferroportin disease is caused by mutations in the iron exporter ferroportin. The clinical characteristics and molecular basis of the various types of non-HFE haemochromatosis are reviewed. The study of these disorders and the molecules involved has been invaluable in improving our understanding of the mechanisms involved in the regulation of iron metabolism.

Taxonomic Studies on a New Marine Ciliate, Apocoleps magnus gen. nov., spec. nov. (Ciliophora, Colepidae), Isolated from Qingdao, China

The morphology and infraciliature of a new marine colepid ciliate, A＇pocoleps magnus gen. nov., spec. nov., are de- scribed based on living observations and silver impregnations. The new genus Apocoleps is characterized by having 8 （vs. 6 in most other related genera） armour tiers, spines at both ends of the cell, 3 adoral organelles and plates with 4 reniform uni-windows. Apo- coleps magnus spec. nov. is defined by the following features： body elongated and slightly curved, about 100-120p.mx 3545 lam in vivo; anterior tertiary tier plate with four uni-windows, most secondary and main tier plates with four uni-windows, posterior tertiary tier plate with two uni-windows; left plate margin slightly serrated; on average 23 transverse and 22 longitudinal ciliary rows; one terminal contractile vacuole; marine habitat.

Dietary Leucine Requirement of Juvenile Japanese Seabass(Lateolabrax Japonicus)

A 56-day feeding trial was conducted to examine the dietary leucine requirement of juvenile Japanese seabass in sea- water floating net cages （1.5 m × 1.5 m × 2.0 m）. Six isonitrogenous （crude protein 40%） and isoenergetic （gross energy 20 kJ g-1） diets were formulated to contain different concentrations of leucine （0.9%, 1.49%, 2.07%, 2.70%, 3.30% and 3.88% of dry matter）. Crys- talline L-amino acids were supplemented to simulate the whole body amino acid pattern of Japanese seabass except for leucine. Three groups （30 fish individuals each, 8.0g±0.20g in initial weight） were fed to apparent satiation at 5：00 and 17：30 every day. During the experimental period, the water temperature ranged from 26 to 32℃ and salinity from 26 to 30, and the dissolved oxygen was maintained at 7mgL-l. The results showed that weight gain （WG）, nitrogen retention （NR）, feed efficiency （FE） and protein efficiency ratio （PER） were significantly increased when dietary leucine was increased from 0.90% to 2.70% of dry matter, and then declined. WG was the highest when fish were fed D4 containing 2.70% of leucine. No significant differences were observed in body composition among dietary treatments （P 〉 0.05）. Considering the change of WG, the optimum dietary leucine requirement of juve- nile Japanese seabass was either 2.39% of dry matter or 5.68% of dietary protein.

Proteinases from the digestive organs of black carp （Mylopharyngodon piceus）： Partial characterization and protein digestibility in vitro

A series of experiments based on electrophoretical and biochemical assays were conducted to partially characterize proteinases present in the hepatopancreas and intestine of black carp （Mylopharyngodon piceus）, and investigate enzymatic activity and protein digestibility in vitro. Casein digestion assays revealed the presence of acidic proteinases with optimum activity in the range of pH 2.0-2.5 and alkaline proteinases with significantly higher activities both in the range of pH 8.1-8.6 and near pH 9.5. The inhibition and substrate specificity assays showed that trypsin and chymotrypsin are the main active components of the alkaline proteinases. The SDS-substrate-PAGE showed that the crude extract of black carp intestine had eight types of alkaline proteinases with the molecular mass range of 27.5-78.5 kDa while the crude extract of black carp hepatopancreas had six types of alkaline proteinases with the molecular mass range of 27.5-78.5 kDa. These enzymes were characterized as trypsin （27.5 kDa, 30.1 kDa）, chymotrypsin （40.5 kDa, 42.5 kDa）, serine proteinases （32.1 kDa, 33.2 kDa） and non-serine proteinase （61.5 kDa, 78.5 kDa）.In vitro protein digestibility assays showed that black cardcan be able to utilize a wider range of proteins.

Protein-sparing effect of carbohydrate in diets for juvenile turbot Scophthalmus maximus reared at different salinities

The aim of the present study was to investigate the protein-sparing effect of carbohydrate in diets for juvenile turbot(Scophthalmus maximus) reared at five salinities(12,18,24,30,and 36).The fish were fed three isocaloric and isolipidic diets for 60 days.The results show that specific growth rate(SGR)and feed conversion efficiency(FCE) were higher in fish reared at salinities of 18 and 36,but lower at 12.Fish fed with diet C25P40(25%carbohydrate and 40%protein) had lower SGR and FCE values compared with those fed with the C5P52(5%carbohydrate and 52%protein) and C15P46(15%carbohydrate and 46%protein) diets;however,there was no statistical difference between diet C5P52 and C15P46.SGR and FCE values were unaffected by diet composition in fish reared at salinity 36.Hepatic lipogenic enzyme activities were higher in fish reared at 18 and 36,but lower at 12,while glucokinase(GK) activity was higher in fish reared at 12,and lower at 18 and 36.Dietary starch enhanced GK activity while depressing lipogenic enzyme activity.However,lipogenic enzyme activity increased with increasing dietary starch in fish reared at 36.It is recommended that salinity should be maintained > 12 in the farming of juvenile turbot.In addition,an increase in gelatinized starch from 5%to 15%could spare 6%dietary protein in fish reared at salinities of 18-30,while higher salinity(36) could improve dietary carbohydrate use and enhance the protein-sparing effect,which is linked with the induction of lipogenic capacities.

Expression and characterization of Mac-1-FP fusion protein in CHO cells

Objective: To construct mammalian cell expression vectors for Mac-1 with CFP and YFP and apply FRET to study the dimerization and function of CD11 b( Mac-1 α subunit) and CD18(Mac-1 β subunit). Methods: The mammalian cell expression vector for CD11b fused with CFP at the carboxyl terminal was constructed to create recombinant plasmid of pCD11b-CFP. Then pCD11b-CFP was co-transfected with pYFP-CD18 into CHO cell, a fibroblast like cell line, as a target cell within which there are some signal pathways involved in inflammatory stimulation but without endogenous Mac-1. Then CHO cells stably expressing both CD11b-CFP and YFP-CD18 fusion proteins were selected by Western blot and laser scanning confocal microscope. Results: The cyan and yellow fluorescence in co-transfected positive CHO cells were observed under a fluorescence microscope. CHO-Mac-1-FP cells stably expressing both CD11b-CFP and YFP-CD18 fusion proteins were obtained as demonstrated by Western blot successfully. The adhesive activity of CHO-Mac-1-FP cells with CHO-1CAM-1 cells was increased markedly by treatment with PMA, suggesting the translocation of GD11b-CFP and YFP-CD18 to the plasma membrane in CHO-Mac-1-FP cells and dimerization of CD11b-CFP and YFP-CD18 just as the function of the wild type Mac-1. Conclusion: CHO-Mac-1-FP cells with adhesive activity are established successfully, thus CHO-Mac-1-FP cells may be useful for the study of Mac-1 by FRET and for other purposes.