Simultaneous Determination of Hexoestrol,Diethylstilbestrol, Estrone and 17-Beta-estradiol in Feed by Gas Chromatography-mass Spectrometry

A method was developed for the simultaneous determination of four kinds of estrogens(hexoestrol, diethylstilbestrol, estrone, and 17-beta-estradiol) in feed by gas chromatography-mass spectrometry(GC-MS). After the sample was extracted by ethyl ether and cleaned-up on HLB phase extraction column, four kinds of estrogens were derived and quantified in gas chromatographymass spectrometry. The results showed that the linear detectable ranged from 2.5 ng·mL~(-1) to 250 ng·mL~(-1) for hexoestrol and from 5 ng·mL~(-1) to 500 ng·mL~(-1) for three other estrogens with the correlation coefficients(R~2) were no less than 0.990. The recoveries were in the range of 76.34%-96.33% and the relative standard deviation was no more than 22.7%. The limits of quantitation(LOQ) for all analytics were between 10 ug·kg~(-1) and 20 ug·kg~(-1).The method was accurate and sensitive and could meet the actual requirements for the analyses of feed samples.     

中医药通过调节Th17细胞分化在自身免疫性炎症性疾病中作用的研究进展

辅助性T淋巴细胞17（T helper 17,Th17）与多种炎症性疾病、自身免疫性疾病及肿瘤密切相关。在自身免疫性炎症性疾病中大多存在偏向Th17一侧的分化失衡,Th17细胞的分化程度往往决定了炎症程度及持续时间。中医药在治疗炎症性疾病方面积累了丰富的经验,具有一定的优势和潜力,许多中药及中药复方被证实有明显的免疫调节及抗炎作用。本文就Th17细胞的分化调节及中医药在类风湿性关节炎、溃疡性结肠炎、强直性脊柱炎及慢性前列腺炎等疾病中对Th17细胞的调节作用研究进展进行综述。     

香鱼白细胞介素17C基因克隆及鳗利斯顿氏菌侵染后的时空表达

为了解香鱼白细胞介素17(interleukin 17,IL-17)的序列特征、系统发育及其与鳗利斯顿氏菌侵染的相关性,实验采用RACE-PCR方法扩增出香鱼IL-17C基因c DNA序列,并通过实时荧光定量PCR(q RT-PCR)技术检测了鳗利斯顿氏菌侵染后香鱼各组织中IL-17C基因的表达变化。结果显示,香鱼IL-17C基因c DNA序列全长1 087 bp,含534 bp的开放阅读框,在3′端非编码区存在7个m RNA不稳定信号(ATTTA)。预测该基因编码177个氨基酸,N端19个氨基酸为信号肽序列,成熟蛋白中含有8个半胱氨酸残基。系统进化树分析显示不同物种的相同IL-17亚型聚为一支,香鱼IL-17C与其他脊椎动物IL-17C聚为一支;同源比对结果显示香鱼IL-17C与虹鳟的IL-17C1和IL-17C2亲缘关系最近,相似性分别为40.72%和33.51%。q RT-PCR检测显示IL-17C基因在健康香鱼的心、肝、脾、头肾、鳃、脑、肌肉和肠等组织中均有表达,鳃和头肾中表达量较高。经鳗利斯顿氏菌侵染后,香鱼肝脏中IL-17C基因表达变化最大,菌侵染8 h时的表达量为对照组的9.17倍;其次为脾,菌侵染4 h时的表达量为对照组的6.69倍。研究表明,香鱼IL-17C基因的表达与病原菌侵染密切相关,可能在免疫功能中具有重要作用。     

Improved vitellogenesis,gonad development and egg diameter in catfish (Pangasianodon hypophthalmus) supplemented with turmeric (Curcuma longa) powder

An experiment was conducted to study the effect of supplementing catfish (Pangasianodon hypophthalmus) with turmeric (Curcuma longa) on the vitellogenic capacity of the liver as a new method to improve reproductive performance in oviparous animals. The experimental catfish were assigned to a completely randomized design consisting of four doses of supplemental turmeric (0, 1.2, 2.4 and 4.8 g/kg feed) with 10 catfish with body weights ranging from 2 to 4 kg in each group. Blood samples were collected on days 0, 14, 28, 42 and 56 of turmeric supplementation to measure the concentrations of plasma oestradiol‐17ß and vitellogenin. The weights of the body, gonads and liver were measured on days 0, 28 and 56. The fecundities of the catfish were measured on days 28 and 56 of turmeric supplementation, and egg diameters were measured on days 28 and 42. The results showed that catfish supplemented with turmeric at a dose of 2.4 or 4.8 g/kg feed had higher growth rates, higher plasma oestradiol‐17β and vitellogenin concentrations, and higher gonad somatic index values and egg diameters. This is the first report demonstrating that the hepatoprotective activity of the curcumin in turmeric could be used to improve both vitellogenin synthesis, which improves nutrient deposition in the ovulating eggs, and the reproductive performance of teleost fish and oviparous animals.     

尼罗罗非鱼cyp17a2生物信息学和雌雄性腺表达差异分析

[目的]进一步研究尼罗罗非鱼cyp17a2的编码蛋白的P450c17-Ⅱ的生物学特征以及在性腺中的表达特点。[方法]根据NCBI网站上登录的尼罗罗非鱼的cyp17a2基因全编码序列,利用相关的生物信息学分析软件分析cyp17a2基因编码的P450c17-Ⅱ的理化性质和结构特点,并分析该基因在不同性腺中的表达。[结果]罗非鱼cyp17a2基因编码的多肽含有521个氨基酸,p I为8.09,脂溶指数为98.33,是不稳定蛋白。罗非鱼cyp17a2基因的二级结构以不规则卷曲所占比例最大,为58.16%,α-螺旋占22.07%。P450c17-Ⅱ是二次跨膜蛋白,有众多蛋白质翻译后修饰位点。在60日龄性腺中,cyp17a2在精巢中的表达量显著高于卵巢。[结论]cyp17a2在尼罗罗非鱼不同性腺中的差异表达和编码P450c17-Ⅱ的生物信息学分析为研究cyp17a2在繁殖过程中的作用奠定了基础。     

A novel Tos17 insertion upstream of Hd1 alters flowering time in rice

Heading date 1 (Hd1) is one of the major determinants of flowering time and regional adaptability in rice (Oryza sativa L.), because there is a significant correlation between Hd1 allelic diversity and phenotypic differences in rice cultivars. In this study, we isolated a novel rice mutant with a Tos17 retrotransposon insertion upstream of Hd1. The Tos17 insertion mutant of Hd1 [designated as Tos17(Hd1)] had decreased photoperiod sensitivity and mRNA expression of Hd1, as compared with wild‐type Nipponbare. Between Tos17(Hd1) and Nipponbare, expression levels of OsPRR1 and OsGI were similar, whereas those of two florigen genes Hd3a and RFT1, were significantly different under both short and long‐day conditions. Tos17(Hd1) showed photoperiod sensitivity intermediate between that of Nipponbare and a near‐isogenic line having a non‐functional allele of Hd1, indicating that Tos17(Hd1) has an attenuated but functional Hd1 allele. Our results revealed that allele mining of Hd1 may provide good opportunities to develop novel rice cultivars showing different levels of photoperiod sensitivity.     

在含等量维生素E饲料中添加β-胡萝卜素和虾青素对仿刺参幼参抗应激能力的影响

在水温11．0～20．0℃、盐度35‰和pH值7．5的条件下,将初始平均体质量为3．46g的仿刺参（Aposticho-pusiaponicusSelenka）幼参放养在容水300L的塑料水槽中,A组投喂含90mg·kg-。β-胡萝卜素＋60mg·kg-1维生素E（VE）的饲料,B组投喂含60mg·kg-1虾青素和60mg·kg-1VE的饲料,以不添加上述3种物质的基础饲料作对照（C组）。饲养80d后,将A、B,及C组幼参直接放人盐度（用淡水和海水或加入海水晶,配制成盐度为0、5、10、15、20、25、30、35、40、45,及50）、氨（用NH4Cl配制成氨浓度（Nn3-N）0、0.5、1．0、2．0、4．0、8．0,及10mg·L-1）、温度（0、4、10、15、20、25、30、35,及40℃）急剧变化的1500mL的白色聚乙烧杯中,每个处理3个重复,测定存活率和体腔液中SOD活力等指标。应激处理期间,不投喂,24h换水一次并吸出排泄物。96h后,盐度、氨处理组的仿刺参存活率高于对照组,存活仿刺参体腔液中SOD活力显著低于对照组（P〈0．05）;但A、B组仿刺参在低温应激时,存活率与对照组差异不显著（P〉0．05）。结果表明：饲料中添加90mg·kg-1β-胡萝卜素＋60mg·kg-1VE或60mg·kg-1虾青素＋60mg·kg-1VE均能显著提高仿刺参对盐度和氨应激的抵抗能力（P〈0．05）,但抗低温应激的效果不显著（P〉0．05）。     

弓形虫棒状蛋白17基因的克隆与序列分析

为研究弓形虫棒状体蛋白17(ROP17)基因的遗传变异,本研究以弓形虫RH株、PRU株和TgC7株为研究对象,首次PCR扩增了其ROP17基因部分序列,将PCR产物纯化后克隆到pMD18-T并测序。将测定的序列与网上下载的弓形虫VEG株、GT1株和ME49株相应序列进行比对,然后用Mega 5.0程序的NJ法和Puzzle 5.2程序的ML法构建系统发育树。结果表明,3株弓形虫分离株的ROP17基因部分序列长度均为1375 bp,A+T含量在49.53%~50.04%之间,3个虫株相应序列的变异碱基数皆小于20个,其变异率为2.3%,氨基酸序列变异率在0~6.11%之间。系统发育结果显示, ROP17基因部分序列能区分弓形虫基因Ⅰ型和Ⅱ型的虫株。本研究结果为进一步研究弓形虫ROP17基因的变异及研制弓形虫ROP17基因的亚单位疫苗提供了理论依据。     

长白猪雌激素受体和卵泡刺激素β基因多态性对繁殖性能的影响

本研究以84头长白猪为试验材料,采用PCR-RFLP方法检测雌激素受体(estrogen receptor,ESR)和卵泡刺激素β(follicle-stimulating hormone β,FSHβ)基因的多态性,并进一步分析这2个基因多态位点及其合并基因型对繁殖性能的影响。结果表明,ESR和FSHβ基因均存在AA、AB和BB 3种基因型,2基因座均处于Hardy-Weinberg平衡状态;单基因效应分析结果表明,ESR基因AB基因型个体部分繁殖性能指标显著高于AA基因型,AB基因型为ESR基因的有利基因型。初产母猪FSHβ基因中BB、AA、AB基因型个体的所有繁殖性能指标均呈递增趋势,而经产母猪中AA基因型个体的性能指标较高,但是差异不显著(P>0.05);ESR与FSHβ的合并效应分析结果表明,基因聚合效应并不是各个基因效应的简单相加,在繁殖性能已经稳定的第3胎次,AA-AB型个体的繁殖性能显著低于其他合并基因型个体,表明该基因型为不利的合并基因型,故在长白猪选种选育过程中应优先考虑被淘汰。     

反相HPLC纯化MAPK去磷酸化肽段研究

[目的]制备含有碳碳双键结构的MAP激酶十三肽。[方法]以化学合成的2种磷酸化MAP激酶保守区段（十三肽）为原料,采用生物化学方法制备含有碳碳双键的十三肽;并确定合适的条件,使用反相液相色谱对这2种多肽进行分离、纯化。[结果]制备得到2种含有碳碳双键的MAP激酶多肽各20 mg。[结论]使用反相HPLC方法分离和制备丝氨酸去磷酸化的这2种多肽是可行的。