市售纯牛奶乳清雌二醇和孕酮含量分析

目的：初步调研市售纯牛奶雌二醇(E2)和孕酮(P4)含量现状。方法：收集同季节6 个品牌(A、B、C、D、E 和F 品牌,各品牌随机采集6 个批号)共36 个批号市售纯牛奶,采用免疫化学发光分析法检测经硫酸酯酶水解的乳清总E2(TE2)和P4。结果：6 个品牌乳清TE2 差异无统计学意义;F 品牌乳清P4 高于C 品牌(P ＜ 0.05),其余各品牌间差异均无统计学意义。36 个批号中,乳清TE2 和P4 最大值与最小值之比分别为2.32 和6.67。A、B、C、D、E 和F 品牌批号间乳清TE2 的最大值与最小值之比分别为1.38、1.60、1.82、1.79、2.17 和1.68,其P4的最大值与最小值之比分别为3.19、5.98、2.89、1.38、1.86 和1.67。6 个品牌的乳清TE2 和P4 均值的最大值与最小值之比分别为1.25 和3.05。结论：市售纯牛奶品牌间乳清P4 存在差异,且同品牌不同批号波动较大;批号间乳清TE2 也有一定差异。     

气质联用法检测养殖虾中的5种雌激素残留

建立一种加速溶剂萃取-气质联用检测养殖虾中的5种雌激素(雌二醇、己烯雌酚、雌酮、炔雌醇、雌三醇)残留的分析方法。采用加速溶剂萃取仪对养殖虾进行提取、净化,选用乙酸乙酯进行洗脱,氮吹仪浓缩至近干,硅烷化试剂N-甲基三甲基硅基三氟乙酰胺(MSTFA)-三甲基硅烷(TMCS)(99︰1,V/V)衍生后进样气相色谱质谱仪定性定量检测。结果显示,5种雌激素质量浓度在0.01~1.0μg/mL范围内呈良好的线性关系,相关系数(R2)均大于0.995;5种雌激素检出限在0.09~0.14μg/kg之间,低(1μg/kg)、中(5μg/kg)、高加标量(10μg/kg)三个不同浓度水平下的加标率在80.6%~93.9%之间,相对标准偏差(RSD)在1.67%~4.72%之间(n=6)。该方法操作简单,灵敏度与准确度高,适用于养殖虾中雌激素的检测。     

Evaluation of methods of resynchronization for insemination in cows of unknown pregnancy status

The objectives of this study were to evaluate the effect of 3 methods of resynchronization of estrus and ovulation for lactating dairy cows of unknown pregnancy status on conception rate and time to pregnancy. Holstein cows (n = 495) were randomly assigned to 1 of 3 treatments: 1) control (n = 167), resynchronization with a timed AI protocol upon diagnosis of nonpregnancy on d 31 after preenrollment AI (PAI); 2) CIDR-G (n = 159), use of an intravaginal progesterone insert from d 14 to 21 after AI, with AI at estrus from d 21 to 24 and initiation of a timed AI protocol on d 24 after AI in cows not reinseminated; 3) CIDR-G + ECP (n = 169), the same treatment as CIDR-G but with an injection of 1 mg of estradiol cypionate at the time of progesterone insert removal. Cows were continuously reenrolled in the same treatment until diagnosed as pregnant, which resulted in a total of 1,148 AI (495 PAI and 653 resynchronized AI; RAI). Blood was collected from 1,001 cows on d 14, 21, and 24 after each AI for analysis of progesterone, and ovaries were scanned on d 21, 24, and 31 after AI. The presence of an active corpus luteum was presumed based on progesterone ≥1 ng/mL. Pregnancy was diagnosed by ultrasonography on d 31 and 61 after AI. The presence of an active corpus luteum and the incidence of luteolysis were similar for all treatments from d 14 to 24; however, luteolysis increased in the CIDR-G + ECP treatment from d 21 to 24. Conception rates for the PAI and all AI were similar on d 31 and 61 after insemination. Conception rates at 31 and 61 d after the RAI were also similar among treatments. Overall pregnancy loss for the PAI, RAI, and all AI were similar for all treatments. The accuracy of estrous detection, based on progesterone concentration within 2 d of detection of estrus, was similar for all treatments for the RAI and averaged 95.3%. The estrus-detection rate (EDR) decreased for the CIDR-G and CIDR-G + ECP treatments from d 14 to 21, but increased from d 21 to 24 compared with control cows; however, the EDR was smaller for cows in the CIDR-G treatment during the entire resynchronization period compared with those in the CIDR-G + ECP and control groups. The reinsemination interval was reduced in cows receiving the CIDR-G + ECP treatment compared with control cows because of increased EDR after removal of the intravaginal insert; however, the interval from study enrollment to pregnancy was not different among treatments. These results indicate that the reproductive performance of dairy cows did not differ among the 3 resynchronization treatments evaluated.     

Reproductive performance of anestrous dairy cows treated with progesterone and estradiol benzoate

Anestrus is a major reproduction problem in pasture-based dairy operations that results in poorer reproductive outcomes than herdmates detected in estrus before the start of the seasonal breeding program. The objective of the current study was to assess a combined progesterone and estradiol benzoate treatment program including resynchrony with no treatment. Anestrous pasture-fed dairy cattle (n = 756) in 9 herds were blocked by herd and age and assigned within sequentially presented pairs of cows to be treated with an intravaginal progesterone-releasing insert for 8 d plus 2 mg of estradiol benzoate injected i.m. at insert insertion and 1 mg of estradiol benzoate injected 1 d after insert removal (d −1). Those cows detected in estrus from 0 to 3 d had a used progester-one-releasing insert reinserted for 6 d commencing on d 16 with 0.5 mg of estradiol benzoate injected i.m. 1 d after insert removal (treatment). The other cow within the pair was left as an untreated control (control). Treatment increased the risk of insemination and pregnancy by 28 d into the breeding program and resulted in conception 15 d earlier compared with controls. In contrast, treatment did not increase the risk of pregnancy after 56 d into the breeding program or at the end of the breeding season. It is concluded that treatment of anestrous dairy cattle with progesterone and estradiol benzoate combined with reinsertion of the progesterone insert resulted in earlier conception, but no difference in the final pregnancy rate compared with no treatment.     

液相色谱-串联质谱法测定动物肌肉和牛奶中的雌二醇

目的 建立丹磺酰氯衍生液相色谱-串联四极杆质谱法检测鸡肉、牛肉、牛奶中雌二醇的高灵敏分析方法.方法 酶解后的样品用HLB和氨基固相萃取柱富集净化,氮气吹干后,在60℃条件下丹磺酰氯衍生5 min;以乙腈-水(含0.1％乙酸)为流动相,采用多离子反应监测正离子模式进行检测.结果 鸡肉、牛肉、牛奶中雌二醇的检出限为0.6 ...     

Cellular proliferation rate and insulin-like growth factor binding protein (IGFBP)-2 and IGFBP-3 and estradiol receptor alpha expression in the mammary gland of dairy heifers naturally infected with gastrointestinal nematodes during development

Mammary ductal morphogenesis during prepuberty occurs mainly in response to insulin-like growth factor-1 (IGF-1) and estradiol stimulation. Dairy heifers infected with gastrointestinal nematodes have reduced IGF-1 levels, accompanied by reduced growth rate, delayed puberty onset, and lower parenchyma-stroma relationship in their mammary glands. Immunohistochemical studies were undertaken to determine variations in cell division rate, IGF-1 system components, and estradiol receptors (ESR) during peripubertal development in the mammary glands of antiparasitic-treated and untreated Holstein heifers naturally infected with gastrointestinal nematodes. Mammary biopsies were taken at 20, 30, 40, and 70 wk of age. Proliferating cell nuclear antigen immunolabeling, evident in nuclei, tended to be higher in the parenchyma of the glands from treated heifers than in those from untreated. Insulin-like growth factor binding proteins (IGFBP) type 2 and type 3 immunolabeling was cytoplasmic and was evident in stroma and parenchyma. The IGFBP2-labeled area was lower in treated than in untreated heifers. In the treated group, a maximal expression of this protein was seen at 40 wk of age, whereas in the untreated group the labeling remained constant. No differences were observed for IGFBP3 between treatment groups or during development. Immunolabeling for α ESR (ESR1) was evident in parenchymal nuclei and was higher in treated than in untreated heifers. In the treated group, ESR1 peaked at 30 wk of age and then decreased. These results demonstrate that the parasite burden in young heifers negatively influence mammary gland development, affecting cell division rate and parameters related to estradiol and IGF-1 signaling in the gland.     

基于表面等离子共振的全自动浓度检测系统

研制了以SpreetaTM传感器为核心,结合液体处理系统、信号检测与控制器、计算机分析处理软件等模块的小型表面等离子共振（SPR）检测系统.运用该系统,通过在传感器表面偶联牛血清蛋白（BSA）,采用直接检测法测量了不同浓度的BSA抗体溶液,得到了BSA抗体溶液的SPR传感图,建立了BSA抗体的浓度标准曲线.在传感器表面固定雌二醇（E2）,采用间接检测法测量了不同浓度的E2溶液,根据E2溶液SPR响应信号的变化量,建立了E2的浓度标准曲线.实验中系统的分辨率为10-5折射率单位（RIU）.对于实际的待测样品,只要测得其SPR信号的变化量,结合标准曲线就能够得出样品的浓度.     

Structure-activity relationships for a family of benzothiophene selective estrogen receptor modulators including raloxifene and arzoxifene

The search for the "ideal" selective estrogen receptor modulator (SERM) as a substitute for hormone replacement therapy (HRT) or use in cancer chemoprevention has focused on optimization of estrogen receptor (ER) ligand binding. Based on the clinical and preclinical benzothiophene SERMs, raloxifene and arzoxifene, a family of SERMs has been developed to modulate activity and oxidative lability. Antiestrogenic potency measured in human endometrial and breast cancer cells, and ER ligand binding data were correlated and seen to provide a guide to SERM design only when viewed in toto. The in vitro studies were extended to the juvenile rat model, in which the desired antiestrogenic profile and putative cardiovascular benefits of SERMs were observed.     

HPLC-fluorescence detection and adsorption of bisphenol A, 17beta-estradiol,and 17alpha-ethynyl estradiol on powdered activated carbon

The adsorption of three estrogenic compounds (bisphenol A (BPA), 17beta-estradiol (E2), and 17alpha-ethynyl estradiol (EE2)) on several powdered activated carbons (PAC) was investigated. Without preconcentration, method detection limits (MDL) using high-performance liquid chromatography (HPLC) with fluorescence detection at an excitation wavelength of 280 nm and an emission wavelength of 310 nm were 0.88, 1.15, and 0.96 nM for BPA, E2, and EE2, respectively. Compound recoveries were >90% in raw drinking water matrices. PAC screening studies (six PAC brands) indicated all three compounds were removed by PAC, but the percentage removal ranged from 31% to >99% based upon PAC type/dosage and presence/absence of natural organic matter. The order of removal (E2>EE2>BPA) corresponded with logK(ow) values for the compounds (3.1-4.0, 3.7-3.9, 3.3, respectively). Kinetic and PAC dose-response experiments were conducted with the two best performing PACs. Increasing contact time and PAC dose improved compound removal. Freundlich isotherm parameters were fit to the experimental data. This study confirms that PAC treatment is feasible for >99% removal of three estrogenic compounds from raw drinking waters that may be at risk for containing such compounds, at least at initial concentration of 500 ng/l and higher.