Protective Effect of Wheat Peptides Against Small Intestinal Damage Induced by Non-Steroidal Anti-Inflammatory Drugs in Rats

Non-steroidal anti-inflammatory drugs（NSAIDs） were able to produce tissue damage and oxidative stress in animal models of small intestinal damage. In this study, the putative protective effect of wheat peptides was evaluated in a NSAID-induced small intestinal damage model in rats, different doses of wheat peptides or distilled water were administered daily by intragastric administration for 30 d until small intestinal damage was caused. Before sacrificing, NSAIDs（aspirin and indomethacin） or physiological saline were infused into the digestive tract twice. Wheat peptides administration reduced edema and small intestinal damage, and significantly decreased the level of tumor necrosis factor（TNF）-α in mucous membrane of small intestine. Oxidative stress was significantly increased after NSAID infusion and was reduced by wheat peptides. Wheat peptides increased glutathione peroxidase（GSH-Px） activity in mucous membrane of small intestine. μ-Opioid receptor mRNA expression decreased more significantly in wheat peptides treated rats than in the model control group. Overall, the results suggest that non-steroidal anti-inflammatory drugs induced small intestinal damage in rats and wheat peptides administration may be an effective tool for protecting small intestinal tissue against NSAID-induced small intestinal damage and oxidative stress.     

泥鳅消化道6种内分泌细胞的免疫组织化学定位(英文)

[Objective] The study was to immunohistochemical localize the endocrine cells from digestive tract of Misgurnus anguillicaudatus.[Method] Using six gastrointestinal hormone antisera, the endocrine cells from digestive tract of M.anguillicaudatus was immunohistochemical localized.[Result] The 5-hydroxytryptamine immunoreactive(5-HT-IR) cells distribute in oesophagus, foregut and midgut; the distribution density was determined to be forepart of foregut> oesophagus and hindpart of foregut>gut, and the differences in the three density gradients reached significant level. Like PP-IR, SS-IR cells were observed mostly in oesophagus, followed by hindpart of foregut, least in forepart of foregut, but never found in gut and hindgut. The three kinds of immunocompetent cells Gas-IR, Glu-IR and SP-IR were not detected in each part of digestive tract. [Conclusion] This study may provide basic data for studying the nutritional and digestive physiology, as well as the preparation of meridic diets for M.anguillicaudatus.     

Immune Responses of Chicken Immunized with Marek‘s Disease Vaccines

The experiment was conducted to study the dynamic changes of immune responses of chicks immunized with Marck‘s disease(MD) trivalent vaccine and turkey herpesvirus (HVT)at one day age.Results were found that after immunization of chicks with MD vaccines,the interlcukine-2 (IL-2) inductive activity and IL-2 receptor expression of T cells from thymus and spleen significantly increased.suggesting that the immunoregulative function was markedly enhanced in the immune organs;the number of antibody-producing cells,the number and proliferative function of T cells rose markedly in Bursa Fabricius,Splcen and thymus,indicating that the ccllular and humoral immune responses were elevated remarkablly in the central and peripheral immune organs; the number of T and antibody-producing cells as well as the content of IgG,IgA and IgM obviously mounted in cecal tonsil,Harder ian gland mucosal lymphoid tissucs of bronchus along with tears,trachea washings,bilc and intestinal fluids,demonstrating that the local and mucosal immunity was raised in the respiratory and digestive tract;the levels of immune responses mentioned above in the trivalent vaccine-immuniacd chicks were apparently higher than those of HVT-immunized birds.     

The SNPs Analysis of the Exons of Three Genes of MyoD Gene Family in Seven Swine Breeds （Line）

The study objects includes seven swine breeds： Minzhu, Sanjiangbaizhu, Yorkshire, Landrace, Junmuyihao, Duroc and Double muscle Yorkshire. According to the sequences of MyoG, MyoD and Myf5 of swine in GenBank, seventeen pairs of primers for MyoG, MyoD and Myf5 were designed. PCR-SSCP technology was applied to detect SNPs of the exons of the three genes. The results showed that no polymorphism was in MyoG and MyoD, and some SNPs were in three exons of Myf5. There was one mutant site in the first exon of Myf5 （G → C）, three mutant sites in the second exon of Myf5 （C → A, A → G and G → A）; in the third exon of Myf5, there was one base A deficiency at 3 387 bp, three bases T deficiency at 3417 bp, one mutant site at 3443 bp （T → C）. This study obtained a tendency conclusion that gene frequency of allele M of Myf5 on the one hand is positively correlated with lean meat percentage, on the other hand is correlated with the orientation of selective breeding; it also deduced that allele F is possibly correlated with high lean meat percentage. Through statistical analysis, allele A, B, C of Myf5 have no obvious correlation with lean meat percentage of different swine breeds. In addition, the high polymorphism of Myf5 showed that seven swine breeds are rich in genetic variation, and have high selective competency.     

Effect of Dietary Alanyl-glutamine Supplementation on Growth Performance,Development of Intestinal Tract,Antioxidant Status and Plasma Non-specific Immunity of Young Mirror Carp（Cyprinus carpio L.）

Exogenous alanyl-glutamine（Aln-Gln） was evaluated for its effects on growth performance, intestinal structure and function, antioxidant status and non-specific immunity of young carp（Cyprinus carpio L.）. Six diets supplemented with 0, 2.5, 5.0, 7.5, 10.0, or 15.0 g · kg-1 of Aln-Gln were fed to fish for 12 weeks. Supplementation with 7.5, 10.0, or 15.0 g · kg-1 of Aln-Gln significantly increased weight gain rate（WGR）, protein efficiency ratio（PER）, but feed conservation rate（FCR） and survival were not affected（P〉0.05）. The intestinal fold height and number, digestive enzyme, Na＋, K＋-ATPase activities was found to be significantly high（P〈0.05） with increasing dietary Aln-Gln supplementation up to 7.5 g · kg-1, but there were no significant differences for Aln-Gln supplementation from 7.5 to 15.0 g · kg-1. The glutathione peroxidase（GPX） activity, glutathione（GSH）, superoxide dismutase（SOD） activity increased and malondialdehyde（MDA） levels decreased significantly（P〈0.05） in the intestine, hepatopancreas, plasma and muscles. The plasma complement-3（C3） and complement-4（C4） levels were significantly（P〈0.05） improved at 5.0 g · kg-1 level and decreased when over 7.5 g · kg-1. The plasma lysozyme（LSZ） activity increased significantly（P〈0.05） at 7.5, 10.0, or 15.0 g · kg-1 level. In summary, the results showed that Aln-Gln improved growth performance, development and function of the intestine, the activity of the antioxidant defense system and the plasma non-specific immunity of the carps. The optimal Aln-Gln level was 8.24 g · kg-1 diet for WGR based on broken-line regression model analysis.     

Development of Alfalfa （Medicago sativa L.） Regeneration System and Agrobacterium-Mediated Genetic Transformation

The regeneration ability of four alfalfa （Medicago sativa L.） cultivars, Xinjiang Daye, Longdong, Gannong 1 and Gannong 3, was studied, and the effects of various cultivars, explant sources and medium recipes on regeneration were compared. The better callus forming frequency obtained from hypocotyls of Xinjiang Daye is 88.5% and regeneration frequency is 9.8% in our initial experiments. To further optimize regeneration system for genetic transformation, we therefore changed concentrations of plant growth regulators and supplemented with glutamine into callus-induction and shoot-regeneration media. Callus forming frequency and shoot differentiation frequency were increased to 100%. The time taken to generate transgenic plants （16 weeks） was shorter than that for previouse procedure （25 weeks） and regeneration frequency was promoted to 15.1%. The results show that addition of glutamine is particularly important for shortening period of regeneration and promoting regeneration frequency. For study of genetic transformation of alfalfa, Agrobacterium tumefaciens-mediated transformation of Xinjiang Daye was developed based on this optimized regeneration system. The plant expression vector carrying two glutamine synthetases （GS 1 and GS2） and △1-pyrroline-5-carboxylate synthetase （P5CS） gene was used for alfalfa in vitro transformation. Six transgenic alfalfa plantlets with resistance to PPT were obtained. The introduction of foreign genes into plants was assessed in the transformants by PCR analysis and Southern hybridizations.     

Cell Ultrastructure of Kiwifruit （Actinidia chinensis） Shoot Tips During Cryopreservation

The changes in the cell ultrastructure of in vitro cultured shoot tips from dwarf genotype of kiwifruit （Actinidia chinensis Ganmi 5） during cryopreservation were investigated. Shoot tips were preserved in liquid nitrogen using vitrification, and the cell ultrastructure was examined using transmission electron microscopy （TEM）. The regular ultrastructure of the cell wall, cell membrane and nucleus of shoot tips could be damaged during the freezing and thawing associated with preservation using liquid nitrogen. The cell plasmolysis was increased and freezing tolerance was improved after precultufing and dehydrating in a preservation and vitrification solution （PVS2） （30% glycerol （Gly）＋ 15% ethylene glycol （EG）＋ 15% dimethylsulfoxide （DMSO） ＋ 0.4 mol L^-1 sucrose）. The structure of some cells with low degree of injury and reversible damage was similar to that of the control and they could undergo normal cell division and differentiation. Besides, they could recover automatically and regenerate after their reculture.     

Effects of Auxins and Media on Callus Induction of Chinese Spring Wheat （Triticum aestivum L.）

The effects of auxins and media on callus induction from the mature and immature embryos of Chinese spring wheat （Triticum aestivum L.） varieties were investigated. It was found that genotype, medium, auxin source and concentration had the significant effects on the induction of embryogenic callus, explants germination and the increment of callus fresh weight. For immature embryos cultured on MS medium, 2 mg L^-1 of 2, 4-D was optimal, and the highest frequency of embryogenic callus （33.50%） was observed. For the mature embryos on N6 medium, 4 mg L^-1 of 2, 4-D was optimal. The frequency of embryogenic callus and increment of callus fresh weight on 2, 4, 5-T media were higher than those on 2, 4-D media, and in the presence of 2, 4, 5-T the precocious germination of explants for all genotypes were significantly suppressed. These results indicated that 2, 4, 5-T was superior to 2, 4-D and NAA in the culture of immature embryos. This is the first report about the effect of 2, 4, 5-T and NAA on wheat tissue culture, particularly in comparison with 2, 4-D in detail.     

SNP Analysis and Haplotype Identification in Chymotrypsin Inhibitor-2 （CI-2） Gene of Barley

Barley chymotrypsin inhibitor-2 （CI-2） was considered to be a promising candidate for enhancing the nutritional value of other cereals by increasing its concentration as it is rich in lysine than any other storage protein. Also, it was proposed that CI-2 might play an important role in the inhibition of proteolytic enzymes from pests or pathogens as CI-2 can strongly inhibit chymotrypsin and subtilisin. In this study, a total of 93 CI-2 gene sequences were isolated from wild and cultivated barley. 48 SNPs and 4 indels were detected across the entire sequences. The frequency of SNPs in the noncoding region （1 out of 9 bases） was slightly higher than that in the coding region （1 out of 10.7 bases）. In all, 33.3% of the candidate cSNPs resulted in amino acid changes. As a total, the 24 cSNPs resulted in 15 amino acid changes. Ten distinguishable haplotypes were detected, among which 3 haplotypes were shared in the most barley accessions, whereas the rest of the haplotypes appeared at a lower frequency. In addition, three haplotypes （haplotype 4, 8, and 9） were unique for single accessions. These results suggested that low diversity at the CI-2 locus was detected among the cultivated and wild barley.     

Impact of Planting Position and Planting Material on Root Yield of Cassava （Manihot esculenta Crantz）

Cassava （Manihot esculenta Crantz） is a perennial woody shrub with an edible root, which grows in tropical and subtropical areas of the world. In Africa, cassava provides a basic daily source of dietary energy. It plays an important role in food security and incomes of many rural households in the southern Ethiopia. However, information available on production practices of cassava for the region is insufficient. Hence, field experiment was conducted at Awassa Agricultural Research Center for two successive cropping seasons from 2004 to 2006 to investigate the response of cassava to planting position and planting material. The treatments used were three planting positions （slant, vertical and horizontal） and five planting materials （main stem top part, main stem middle part, main stem bottom part, branch stake top part and branch stake bottom part） were combined in factorial arrangement and laid out in randomized complete block design with three replications. The result revealed that root yield was significantly （P 〈 0.05） affected by the interaction effects of the planting position and planting material. The highest yield （25.2 ton ha^-1） was obtained from the main stem top part planted in slant position whereas the least yield （6.5 ton ha^-1） was obtained from main stem bottom part planted in horizontal position. Based on the findings of this study, areas like Awassa with moderate rainfall slant and vertical planting of main stem top and middle parts could be used as planting material.     

罗非鱼和斑点叉尾鮰血清IgM的纯化及兔抗血清的制备

为深入了解罗非鱼和斑点叉尾鮰免疫机理及建立相关免疫检测方法,采用rProtein A Sepharose亲和层析一步法纯化罗非鱼和斑点叉尾鮰血清免疫球蛋白（IgM）,并制备其兔抗血清.结果表明,rProtein A Sepharose亲和层析法可以较好地分离获得高纯度的罗非鱼和斑点叉尾鮰血清IgM,通过SDS-PAGE电泳检测,发现罗非鱼血清IgM重链和轻链分子量分别为88.0、21.0 kDa,斑点叉尾鮰血清IgM重链分子量为101.0 kDa.以纯化的罗非鱼和斑点叉尾鮰血清IgM为抗原,制备其兔抗血清,间接ELISA检测其效价分别为1∶32000和1∶16000;Western blotting检测分析,发现罗非鱼和斑点叉尾鮰的兔抗血清分别在88.0和101.0 kDa附近各出现1条反应条带,说明其兔抗血清具有免疫活性.     

斑点叉尾鮰(Ictalurus punctatus)肝脏表达的抗菌肽-2(LEAP-2)在E.coli中的融合表达

抗菌肽是存在于生物细胞中的一类具有强抗菌作用的天然小分子多肽,它们在机体免疫系统中发挥了重要作用,被认为是抗生素的理想替代品.本文通过RT-PCR法从斑点叉尾鮰(Ictalurus punctatus)的肝脏中分离到抗菌肽-2(LEAP-2)基因,该基因编码94个氨基酸残基组成的多肽;氨基端的信号肽由28个残基组成,成熟肽含有41个残基;对于鱼类和哺乳动物显示高度保守的4个半胱氨酸残基位于多肽的羧基端区域.通过对LEAP-2基因添加EcoR Ⅰ和Hind Ⅲ酶切位点和对pET32a进行双酶切处理,成功构建了"pET32a-IpL-2"重组表达质粒,并转化工程茵E.coli BL21(DE3).在工程菌对数生长后期加入诱导剂IPTG、经25℃培养后,可溶性的"trxA-IpL-2"融合蛋白得到高程度的表达.Tricine-SDS-PAGE和Western blot分析表明,获得了高纯度的目的蛋白.     

水库斑点叉尾鮰网箱微孔增氧高效养殖试验

为了充分挖掘网箱养殖的生产潜力,提高养殖效益,改善养殖水体环境,减少病害发生,将微孔增氧技术运用于斑点叉尾鮰网箱养殖中,试验结果表明:成活率比对照组提高8.54个百分点;产量比对照组提高33.4 kg/m2;纯收益比对照增加231.8元/m2。     

TaqMan实时荧光PCR快速检测斑点叉尾鮰病毒

根据GenBank登录的斑点叉尾鮰病毒株ORF8基因序列,设计引物和Taqman荧光探针,建立了用于检测斑点叉尾鮰(Ictalurus Punctatus)病毒的实时荧光定量PCR方法;对实时荧光定量PCR反应条件进行了优化,评估了该方法的特异性、灵敏度和重复性,并建立了绝对定量方法。特异性试验证实,该方法只能检测到斑点叉尾鮰病毒,与真鲷虹彩病毒、蛙虹彩病毒、锦鲤疱疹病毒、鳜鱼传染性脾肾坏死病毒等多种常见的水生动物病毒没有交叉反应,具有高度的特异性。标准曲线表明,在3.3×10~3.3×107拷贝数之间有较好的线性关系(r=0.998 3),最少可检测到33个拷贝的阳性质粒,敏感度很高。同一样品于试验内及试验间的重复性试验发现,变异系数分别为0.7%和1.2%,重复性较好。该方法的检测时间从核酸抽提到出具结果仅需3h。试验结果表明,采用实时荧光PCR检测斑点叉尾鮰病毒方法具有特异性好、灵敏度高、检测耗时短的特点,并且能对病毒进行准确的定量分析,不仅适合口岸进出口检验检疫的需求,而且可以作为研究该病毒感染过程的有力工具。     

斑点叉尾鮰对3种致病菌灭活菌苗的免疫应答

将经福尔马林灭活的柱状嗜纤维菌(Cytophaga columnaris)、嗜水气单胞菌(Aeromonas hydrophi-la)和叉尾鮰爱德华菌(Edwardsiella ictaluri)作为免疫原,分别接种斑点叉尾鮰(Ictalurus punctatusRafinsque)后,通过测定受免鱼的交叉凝集抗体效价、头肾和血液中吞噬细胞的吞噬活性及用A.hydrophila活菌攻毒的方法,探讨了斑点叉尾鮰对3种灭活菌苗的免疫应答状况。试验结果表明,对斑点叉尾鮰经腹腔注射接种3种灭活菌苗能刺激受免鱼产生较强的免疫应答,受免鱼产生了对3种致病菌的特异性凝集抗体和交叉凝集抗体,与未接种菌苗的对照鱼相比,受免鱼头肾和血液中吞噬细胞的吞噬活性明显上升,活菌攻毒的结果也证明了受免斑点叉尾鮰对C.columnaris,A.hydrophila和E.ictaluri产生了不同程度的相对免疫保护率(RPS)。     

3种鱼类致病菌粗脂多糖(LPS)对斑点叉尾鮰的免疫原性

用温酚法分别从柱状嗜纤维菌(Cytophaga columnaris)、嗜水气单胞菌(Aeromonas hydrophila)和叉尾鮰爱德华菌(Edwardsiella ictaluri)3种鱼类致病菌中提取粗脂多糖(LPS)作为免疫原,分别接种斑点叉尾鮰(Ictalurus punctatusRafinsque)后,通过测定受免鱼的交叉凝集抗体效价、头肾和血液中吞噬细胞的吞噬活性以及对A.hydrophila活菌细胞内的杀菌活性,探讨了3种致病菌的粗LPS对斑点叉尾鮰的免疫原性。结果表明,从3种致病菌中提取的粗LPS对斑点叉尾鮰均具有较强的免疫原性,受免鱼的血清中存在对3种致病菌的交叉凝集抗体,与对照鱼相比,头肾和血液中吞噬细胞对吞噬原的吞噬活性和对A.hy-drophila活菌杀菌活性明显上升。说明在3种供试菌的粗LPS上不同程度地存在3种致病菌的共同保护性抗原。     

嗜水气单胞菌对盐酸多西环素的耐药性获得与消失速率研究

在离体条件下测定了从人工养殖的斑点叉尾蛔患病鱼体上分离的嗜水气单胞菌（Aerowtonas hydrophila）的4个菌株对盐酸多西环素耐药性获得与消失的速率。结果表明：以25℃条件下培养72h为1代,在含有盐酸多西环素的药物培养基中连续传代9次后,盐酸多西环素对A．hydrophila的最小抑制菌浓度（MIC）上升了31倍,MIC由0．05mg／L上升至1．56mg／L;而将已经获得高耐药性的菌株以穿刺法接种在不合药物的BHI（Difco）琼脂培养基中,以4℃条件下保存20d为1代,连续传代5次,供试菌株的耐药性呈现逐渐消失的趋势,至第5次传代后,其MIC由1．56mg／L下降至0．10mg／L。     

斑点叉尾鮰对3种致病菌外膜蛋白(OMP)的免疫应答

利用从嗜水气单胞菌(Aeromonas hydrophila)、柱状嗜纤维菌(Cytophaga columnaris)和叉尾鮰爱德华菌(Edwardsiella ictaluri)中提取的外膜蛋白(outer membrane protein,OMP)作为免疫原,注射接种斑点叉尾鮰(Ictalurus punctatus Rafinsque)后,通过测定受免鱼的交叉凝集抗体效价、肾脏和血液中吞噬细胞的吞噬活性和采用A. hydrophila活菌攻毒方法,比较了3种致病菌OMP对斑点叉尾鮰的免疫原性。试验结果表明:从3种致病菌中提取的OMP对斑点叉尾鮰均具有较强的免疫原性,受免鱼的血清中存在对3种致病菌的交叉凝集抗体,与对照鱼相比,肾脏和血液中吞噬细胞对吞噬原的吞噬活性和对A. hydrophila活菌攻毒的相对免疫保护率(relative percent survival,RPS)明显上升。说明在3种供试菌的OMP上不同程度地存在共同保护性抗原。     

提高冻藏鮰鱼片解冻品质的研究

研究抗冻剂浸渍对-18℃,-30℃和-50℃ 3种冻藏温度的鮰鱼片解冻后的品质和蛋白质变性的影响。结果表明：添加抗冻剂降低了解冻鮰鱼片的pH值,减少解冻后水分流失,维持较低的K值和挥发性盐基氮含量,保持解冻鱼片较好的品质;降低TBARS值,缓解脂肪氧化;保持较高的盐溶蛋白含量和Ca²⁺-ATP 酶活性,减轻解冻鮰鱼片蛋白质冷冻变性的程度。     

斑点叉尾鮰细菌性病原的分离与鉴定

报道从湖北省宜都市和长阳土家族自治县内清江河内养殖网箱中患病斑点叉尾鮰体内和体表分离致病菌的初步鉴定结果。从患病斑点叉尾鮰上共分离19个菌株,经分别采用注射和浸泡法人工感染试验,证明其中11个菌株能导致试验鱼80%~100%的死亡。根据对菌株的形态和生化特性的测定结果,11个菌株中的BH-001、BH-005、BH-009和BH-011等4个菌株属于叉尾鮰爱德华菌(Edwardsiella ictaluri),而BH-002、BH-015、BH-017和BH-019等4个菌株属于嗜水气单胞菌(Aeromonas hydrophila)。而BH-004、BH-013和BH-016等3个菌株尚未能鉴定其种类。