蛋白质营养水平对工蜂初生重和咽下腺发育的影响

利用不同配比的茶花粉和黄豆粉饲料饲喂蜂群,测定后代工蜂的初生重和咽下腺宽度。结果发现,饲喂100％茶花粉与饲喂“75％茶花粉＋25％黄豆粉”的蜂群,所育工蜂的初生重差异不显著,与饲喂100％黄豆粉的蜂群相比,差异极显著;饲喂100％茶花粉与饲喂“75％茶花粉＋25％黄豆粉”的蜂群相比,6日龄、9日龄、12日龄工蜂咽下腺宽度差异均不显著,与饲喂100％黄豆粉的蜂群相比,6日龄、9日龄差异不显著,而12日龄差异显著。本试验结果表明,蛋白质营养水平对工蜂初生重和咽下腺发育均具有显著的影响。     

cFLIP regulates death receptor-mediated apoptosis in an ovarian granulosa cell line by inhibiting procaspase-8 cleavage

More than 99% of follicles in mammalian ovaries undergo atresia, but the mechanisms regulating the strict selection process are still unclear. Granulosa cell apoptosis is considered the trigger of follicular atresia, which occurs in advance of the death of an oocyte. Cellular FLICE-like inhibitory protein (cFLIP), a homologue of procaspase-8 (also called FLICE), is an intracellular anti-apoptotic protein. It is expressed in granulosa cells of porcine ovaries, where its levels decreases during follicular atresia. We hypothesized that cFLIP regulates granulosa cell apoptosis by acting as a pro-survival factor. In the present study, to further reveal the function of cFLIP in granulosa cells, we examined the anti-apoptotic mechanism of cFLIP using KGN, a human granulosa tumor cell line. Fas-mediated apoptosis was induced by co-treatment with anti-Fas antibody (CH-11), which acts as an agonist of Fas-ligand, and cycloheximide (CHX). When cFLIP was stably expressed in KGN cells following transfection of an expression vector, the Fas-mediated apoptosis was inhibited. Suppression of cFLIP by small interfering RNA (siRNA) spontaneously induced cell death. Silencing of cFLIP promoted cleavage of procaspase-8, and the cell death caused by cFLIP siRNA was completely blocked by a caspase-8 inhibitor (Z-IETD-FMK), indicating that cFLIP regulates apoptosis in KGN cells by inhibiting cleavage of procaspase-8. In conclusion, cFLIP is an essential pro-survival factor for granulosa cells, and it prevents granulosa cell apoptosis by inhibiting procaspase-8 activation.     

雌蚕无性克隆系的构建及应用研究

基于雌蚕无性克隆后代全雌、基因型完全一致的遗传特性,在家蚕遗传育种及杂种优势机理研究等方面具有重要价值.本文介绍了10余年来在雌蚕无性克隆性状的遗传特性、无性克隆系的构建及应用方面所取得的主要进展.     

低聚乳果糖的酶法制备研究进展

低聚乳果糖是一种功能性低聚糖是双歧杆菌的有效增殖因子。具有增殖细菌、低热量、低龋齿、降低血液中的胆固醇、改善血脂、促进钙吸收等特殊的生理功能。低聚乳果糖是以乳糖和蔗糖（1：1）为原料,在节杆菌产的β-呋喃果糖苷酶（EC3．2．1．26）催化作用下生成半乳糖基蔗糖即低聚乳果糖。利用微生物来源的β-呋喃果糖苷酶制备低聚乳果糖具有成本低、稳定性高＼提取方便等特点。不同微生物来源的β-呋喃果糖苷酶具有不同的性质,目前应用比较广泛的是节杆菌产的β-呋喃果糖苷酶。国内对低聚乳果糖做了初步研究,还没有工业化生产的报道,因其具有广阔的应用前景。文章综述了低聚乳果糖的性质、制备、分离、检测及其在饲料、食品、医学等方面的应用。     

家蚕中肠上皮细胞增殖和分化的初步研究

通过分析家蚕自幼虫期到蛹期发育过程中肠上皮细胞的增殖与分化情况,鉴定家蚕中肠干细胞的潜在定位。采用苏木精和伊红(Hematoxylin and Eosin,H&E)染色与4’,6-二脒基-2-苯基吲哚(4’,6-diamidino-2-phenylindole,DAPI)染色追踪变态和发育时期家蚕中肠的形态结构及细胞组成变化,结果显示,幼虫经历蜕皮及变态时,中肠形态结构以及中肠上皮细胞组成均发生明显变化:在幼虫每个龄期的盛食期肠壁较薄,眠前期(蜕皮前)明显变厚,至眠期其厚度达到峰值;中肠上皮层存在柱状细胞(CC)、杯状细胞(GC)和再生细胞(RC)3种细胞,3种类型细胞随龄期逐渐增多,其中各龄期柱状细胞持续增多,至眠期达到峰值,靠近基底膜的小细胞在眠前期增多。利用5-溴脱氧尿嘧啶核苷(5-Bromo-2-deoxyUridine,BrdU)和磷酸化组蛋白(Phospho-histone H3,PHH3)免疫荧光染色检测到中肠上皮细胞,尤其是中肠上皮层靠近基底膜处的小细胞在幼虫各龄眠前期的增殖率最高。同时通过BrdU滞留标记实验在中肠上皮层靠近基底膜处发现了BrdU滞留阳性信号。研究结果发现家蚕幼虫蜕皮时中肠上皮层靠近基底膜处的小细胞发生快速增殖,推测这些小细胞中存在着潜在的家蚕中肠干细胞。     

重组质粒pACYC184-hok/sok的构建及稳定性研究

以低拷贝质粒pACYC184为载体,将hok/sok基因插入到载体BamH Ⅰ位点,构建重组质粒pACYC184-hok/sok稳定系统,同时构建hok/sok基因缺失突变重组质粒pACYC184-hok/sok(M).通过对构建的重组菌进行连续传代和对不同代次的重组质粒进行SphⅠ酶切,分析hok/sok基因的引入对宿主细胞生长的影响.结果表明,重组质粒pACYC184-hok/sok在无抗生素筛选压力下连续传代,传至第135代时质粒稳定率仍是100%,且传代前后的质粒SphⅠ酶切图谱没有发生变化,DNA序列测定表明,插入的hok/sok基因没有发生任何突变.突变重组质粒pACYC184-hok/sok(M)传代前后的质粒SphⅠ酶切结果虽没有发生变化,但其相应的重组菌传至第15代,质粒几乎完全丢失.生长曲线的测定结果表明,与舍pACYC184重组菌生长曲线相似,pACYC184-hok/sok重组菌生长较快,而pACYC184-hok/sok(M)重组菌生长缓慢.以上结果表明,含hok/sok稳定系统,其稳定性明显高于无hok/sok稳定系统的质粒pACYC184以及突变质粒pACYC084-hok/sok(M).重组质粒pAqCYC184-hok/sok具有良好的结构稳定性和分离稳定性.     

高效液相色谱荧光检测法同时检测鸡蛋中氟苯尼考及其代谢物氟苯尼考胺残留

建立了高效液相色谱荧光检测法同时检测鸡蛋中氟苯尼考及其代谢物氟苯尼考胺残留的方法.鸡蛋经碱性乙酸乙酯提取,饱和正己烷脱脂,氮吹仪吹干浓缩后用流动相溶解.用Lichrospher C1s柱,以乙腈-磷酸二氢钠溶液(0.01 mol/L,含0.005 mol/L十二烷基硫酸钠和0.1％三乙胺)(35:65,v/v)为流动相,在激发波长224 nm、发射波长285 nm处用荧光检测器检测.氟苯尼考在0.01～10.0 mg/L、氟苯尼考胺在0.002 5～2.5 mg/L浓度范围内,本方法线性关系良好,相关系数分别为0.99 97和0.999 8.当添加水平氟苯尼考为15～500μg/kg、氟苯尼考胺为5～500 μg/kg时,该方法平均回收率分别为87.41％～92.29％和89.01％～95.15％,相对标准偏差分别为3.51％～5.36％和4.28％～5.72％;检测限分别为1.5 μg/kg和0.5 μg/kg(S/N=3);定量限分别为5μg/kg和2 μg/kg(S/N=10).该方法简便、快速、灵敏度高,可同时检测鸡蛋中氟苯尼考和氟苯尼考胺的残留.     

羽毛角蛋白降解的研究进展

羽毛角蛋白是一种硬性蛋白,在动物饲料、复合材料、食品包装和医药制品等领域展现出良好应用前景。但因羽毛角蛋白难溶解和降解的特殊性,一般方法很难获得可溶性角蛋白,酶法相对于传统物理和化学降解法具有反应条件温和、环境污染小和降解产物稳定等优点,是具有很高实用价值的羽毛降解途径。文中重点介绍角蛋白酶对羽毛的降解作用,并概述物理、化学方法的降解原理以及角蛋白的综合利用。     

Study on Expression Difference of IGFBP-5 Gene in Different Tissues of Kazakh and Yanqi Horses

The study aimed to explore the mRNA expression pattern of insulin-like growth factor binding protein-5 (IGFBP-5) gene in different tissues of Kazakh and Yanqi horses.The expression of IGFBP-5 gene in different tissues of heart,liver,spleen,lung,kidney,small intestine,large intestine,cecum,intercostal muscles,longissimus dorsi muscles,brachialis muscle and gluteus in two horses were detected by Real-time quantitative PCR and compared the mRNA expression in the same tissues of two breeds.The results showed that the expression of IGFBP-5 in longissimus dorsi muscle and brachialis muscle of two breeds were significantly higher than other tissues including heart,liver,spleen,lung,kidney,small intestine,large intestine and cecum (P<0.05),and was the lowest in large intestine.The expression of IGFBP-5 in kidney,small intestine,large intestine,cecum,longissimus dorsi muscle and brachialis muscles of Yanqi horse were higher than in the same part of Kazakh horse,and among those in longissimus dorsi muscle and large intestine of Yanqi horse were extremely significantly higher than in the same part of Kazakh horse (P<0.01),and in small intestine and cecum of Yanqi horse were significantly higher than in the same part of Kazakh horse (P<0.05).The test was for further researching the biological function of IGFBP-5 gene,and it could provide a theoretical basis for genetic improvement of production performance of horse in our country.     

一株乳酸片球菌的筛选、鉴定及其固态发酵饲料原料的产酸分析

本研究从猪的新鲜粪便中分离纯化出一株具有较强产酸效果的乳酸菌HF14,综合形态、生理生化、16SrDNA序列分析,初步鉴定该菌株为乳酸菌片球菌(Pediococcus acidilactici)。对HF14固态发酵豆粕、麸皮、玉米粉与稻壳粉等饲料原料的产酸类别进行分析发现,产生的有机酸以L-乳酸与乙酸为主,其中,发酵玉米粉的pH最低,为3.77,L-乳酸与丙酸含量最高,分别为21.91、1.03mg/g;发酵麸皮的pH降幅最大,丁酸含量最高,为0.72mg/g;发酵稻壳粉的乙酸含量最高,为6.15mg/g,L-乳酸含量最低;发酵豆粕的各个指标居中。