菟丝子黄酮对BPA致小鼠睾丸间质细胞分泌雄性激素异常的保护作用

为研究菟丝子黄酮对双酚A(BPA)干扰小鼠睾丸间质细胞激素分泌的缓解作用,以TM3小鼠睾丸间质细胞为对象,采用1μg/mL～500μg/mL的菟丝子黄酮与TM3(小鼠睾丸间质细胞)细胞共同培养24 h,以MTT法检测细胞增殖,确定菟丝子黄酮对TM3细胞的安全剂量。然后采用1μmol/mL～200μmol/mL的双酚A干扰TM3细胞24 h,以MTT法检测细胞增殖,通过TM3细胞的相对存活率,确定BPA最适染毒剂量;最后先用菟丝子黄酮和TM3细胞共同培养4 h后,再用BPA干扰TM3细胞24 h。通过ELISA检测细胞上清液睾酮(T)和黄体生成素(LH)的含量,并分析菟丝子黄酮对BPA干扰小鼠睾丸间质细胞激素分泌的缓解作用。结果表明,250μg/mL～400μg/mL的菟丝子黄酮(等差法)对TM3细胞作用24 h后,有极显著的增殖效果(P<0.01);80μmol/mL的BPA对TM3细胞作用24 h后,细胞存活率为81%,可作为最适染毒剂量;菟丝子黄酮保护4 h后,再用BPA干扰12 h、24 h后,不同浓度的菟丝子黄酮可显著升高TM3细胞睾酮和黄体生成素的水平(P<0.05)。说明菟丝子黄酮对BPA致小鼠睾丸间质细胞睾酮和黄体生成素的分泌异常具有明显的保护作用。     

牛荚膜A型多杀性巴氏杆菌强菌株rpoE蛋白的原核表达及其免疫保护效力研究

为研究牛荚膜A型多杀性巴氏杆菌(PM)强菌株rpoE蛋白的免疫保护性,本研究利用PCR方法扩增牛荚膜A型PM的rpoE基因,构建pET-28a-rpoE重组表达质粒,经诱导表达后SDS-PAGE检测结果显示,表达的rpoE蛋白相对分子量约为20 ku,与理论值一致;western blot结果显示该蛋白具有很好的特异性和反应原性;纯化的rpoE蛋白经皮下免疫BALB/c小鼠,间接ELISA检测融合蛋白的免疫原性,结果显示rpoE蛋白可以诱导小鼠产生较高水平的特异性抗体;PM攻毒后,免疫组保护率为70%。本研究获得的牛荚膜A型PM重组rpoE蛋白具有良好的免疫原性,免疫小鼠后能够提供一定的保护,可以作为研发PM亚单位疫苗的候选抗原。     

玉米赤霉烯酮对小鼠原代培养T淋巴细胞细胞因子分泌的影响

为探讨玉米赤霉烯酮(zearalenone,ZEA)对动物免疫功能的影响及其机理,试验以分离培养的原代小鼠脾脏淋巴细胞材料,研究了ZEA对T淋巴细胞细胞因子分泌的影响。以刀豆蛋白A(Con A,5 mg/L)作为T细胞活化特异性刺激剂,试验设空白对照组(不加Con A)、Con A组(5 mg/L Con A)、不同浓度的ZEA染毒组(Con A+ZEA 10、20和40μmol/L),处理48,72 h后,使用流式液相多重蛋白定量技术检测T细胞分泌IL-2、IL-3、IL-5、IL-6和GM-CSF等细胞因子的含量。结果显示,与空白对照组相比,Con A组细胞在48,72 h时间段5种细胞因子分泌均明显上升;与Con A组比较,染毒48,72 h后,10,20,40μmol/L ZEA染毒组各细胞因子分泌浓度均有极显著下降(P<0.01),并呈剂量-效应关系。结果表明,ZEA可以抑制T淋巴细胞细胞因子IL-2、IL-3、IL-5、IL-6和GM-CSF的分泌,从而影响机体正常免疫反应的进行,降低整体的免疫功能。     

Extracellular cyclic adenosine monophosphate‐dependent protein kinase A autoantibody and C‐reactive protein as serum biomarkers for diagnosis of cancer in dogs

Protein kinase A, a cyclic adenosine monophosphate (AMP)‐dependent enzyme, normally exists within mammalian cells; however, in cancer cells, it can leak out and be found in the serum. Extracellular cyclic AMP‐dependent protein kinase A (ECPKA) has been determined to increase in the serum of cancer‐bearing dogs. However, there have been no reports in the veterinary literature on serum ECPKA autoantibody (ECPKA‐Ab) expression in dogs with cancer. The aim of this study was to evaluate ECPKA‐Ab and C‐reactive protein (CRP) as serum biomarkers for cancer in dogs. ECPKA‐Ab and CRP levels were detected by an enzyme‐linked immunosorbent assay in serum samples from dogs with malignant tumours (n = 167), benign tumours (n = 42), or non‐tumour disease (n = 155) and from healthy control dogs (n = 123). ECPKA‐Ab and CRP levels were significantly higher in the dogs with malignant tumours than in those with benign tumours or non‐tumour diseases, as well as in the healthy controls (P < 0.001, Kruskal‐Wallis test). There was a significant positive correlation between the neoplastic index, which was developed using ECPKA‐Ab and CRP levels, and the presence of cancer in dogs (P < 0.001); the area under the receiver‐operating characteristic curve was estimated to be >0.85 (P < 0.001). In conclusion, ECPKA‐Ab is a potential serum biomarker for a broad spectrum of cancers. Combined measurement of CRP and ECPKA‐Ab levels in serum improves the sensitivity and accuracy of a diagnosis of cancer in dogs.     

Associations Between Oil and Gas Wells and Arthropod and Vegetation Communities in the Southern Plains

Understanding how energy infrastructure affects local biodiversity and soil characteristics is important for informing restoration and management. However, the rapid rate of modern oil and gas development is beyond the limit of current knowledge and mitigation strategies. In a mixed-grass prairie in western Oklahoma, we assessed the presence and directionality of biodiversity and environmental gradients associated with energy development in an observational framework. Specifically, we sampled arthropods, vegetation, soil temperature, and soil moisture on the edge of active oil well pads and at 1 m, 10 m, and 100 m away from the well pad. Though variable, the abundance and biomass of most arthropod orders was lower on the pad and 1 m away compared with 10 m and 100 m away, suggesting that the pad itself negatively influenced arthropods but that these effects were limited in spatial extent. However, vegetation structure and composition varied more extensively. Vegetation height, shrub cover, and warm season grass cover increased sixfold, threefold, and fourfold, respectively, from on the oil pad to 100 m away. Forb cover was 5 × higher at 10 m from the well pad than on the pad, 1 m away, and 100 m away from the pad. Soil surface temperature was lower at sites farther from well pads, but we found no relationship between soil moisture and distance from well pad. Well pad effects on arthropods and soil temperature appear to be limited to the pad itself, though long-term changes in vegetation structure extend significantly beyond the well footprint and demand a better understanding of the effectiveness of restoration activities around well pads.     

Dormant Season Grazing: Effect of Supplementation Strategies on Heifer Resource Utilization and Vegetation Use

Grazing dormant forage under low-input heifer development strategies typically exposes cattle to low-quality forage. Protein supplementation while grazing dormant range can enhance heifer growth and reproductive performance. We examined resource utilization of heifers and the effects of dormant season grazing on residual vegetation characteristics under two supplementation management strategies. Approximately 100 weaned composite heifer calves were randomly selected and placed into one of two supplementation treatments in each of 2 yr, one receiving a free access 62% crude protein self-fed concentrate and the other receiving a daily hand-fed 20% crude protein cake. Grazing occurred from December (2015 and 2016) through March (2016 and 2017). Thirty transects were randomly located within each pasture for measuring vegetation quality and structure before and after grazing. Daily space use and behavior was evaluated for 21 individuals within each treatment using global positioning system (GPS) collars and resource utilization functions. Heifers supplemented with concentrated protein spent more time grazing per day than heifers supplemented with cake (6.92 ± 0.18, 6.24 ± 0.17 h). Relative use by heifers in the cake treatment was negatively related to horizontal distance from the supplement delivery site early to midwinter ($\stackrel{?}{\overline{\beta }}$ = ? 0.41 ± 0.16, ? 0.53 ± 0.17). Both treatments selected grazing locations relative to standing biomass of perennial grasses ($\stackrel{?}{\overline{\beta }}$ = 0.0005 ± 0.00004) and crude protein ($\stackrel{?}{\overline{\beta }}$ = 0.12 ± 0.007). However, resource selection was highly variable among individuals for both supplementation treatments. We found no treatment effects on pre-post grazing differences in residual cover of litter, grass, forbs and shrubs (P > 0.24). However, the time period when grazing occurred had an effect on residual vegetation conditions (P < 0.01). Our results indicate high levels of variability in grazing site selection by heifers, suggesting future research should incorporate individual animal measurements in an attempt to account for individual animal variability.     

滇型杂交粳稻“寻杂29”在水、旱条件下植株性状变化的研究初报

 根据1990～1994年滇型杂交粳稻“寻杂29”水栽和旱种的试验数据和调查资料,从“寻杂29”在水、旱条件下植株性状变化和产量形成的种种差异,论述该品种对不同生态环境的抗逆性和适应能力,初步探索出杂交水稻“寻杂29”旱种的生长发育及其栽培特点,供生产应用参考.     

Cytoplasmic kinases downstream of GPR30 suppress gonadotropin-releasing hormone (GnRH)-induced luteinizing hormone secretion from bovine anterior pituitary cells

GPR30 is known as a membrane receptor for picomolar concentrations of estradiol. The GPR30-specific agonist G1 causes a rapid, non-genomic suppression of gonadotropin-releasing hormone (GnRH)-induced luteinizing hormone (LH) secretion from bovine anterior pituitary (AP) cells. A few studies have recently clarified that protein kinase A (PKA) and phosphorylated extracellular signal-regulated kinase (pERK) might be involved in cytoplasmic signaling pathways of GPR30 in other cells. Therefore, we tested the hypothesis that PKA and ERK kinase (MEK) are important cytoplasmic mediators for GPR30-associated non-genomic suppression of GnRH-induced LH secretion from bovine AP cells. Bovine AP cells (n = 8) were cultured for 3 days under steroid-free conditions. The AP cells were previously treated for 30 min with one of the following: 5000 nM of PKA inhibitor (H89), 1000 nM of MEK inhibitor (U0126), or a combination of H89 and U0126. Next, the AP cells were treated with 0.01 nM estradiol for 5 min before GnRH stimulation. Estradiol treatment without inhibitor pretreatment significantly suppressed GnRH-induced LH secretion (P < 0.01). In contrast, estradiol treatment after pretreatment with H89, U0126 or their combination had no suppressive effect on GnRH-induced LH secretion. The inhibitors also inhibited the G1 suppression of GnRH-induced LH secretion. Therefore, these data supported the hypothesis that PKA and MEK (thus, also pERK) are the intracellular mediators downstream of GPR30 that induce the non-genomic suppression of GnRH-induced LH secretion from bovine AP cells by estradiol or G1.     

Reaction phenotyping of vinblastine metabolism in dogs

Vinblastine is a vinca alkaloid used either as a single agent or in combination therapy for the treatment of canine mast cell tumours and lymphomas. The objective of this study was to determine which isoform of cytochrome P450 enzyme is responsible for the majority of vinblastine metabolism in dogs. A panel of eight recombinant canine cytochrome P450 enzymes (CYP1A1, CYP1A2, CYP3A12, CYP3A26, CYP2B11, CYP2C41, CYP2C21 and CYP2D15) were incubated in vitro with vinblastine. Findings were confirmed by the use of canine polyclonal antibodies of cytochrome P450 enzymes (CYP1A1, CYP3A12, CYP2B11 and CYP2C21) that were pre‐incubated with individual and pooled hepatic microsomes that were purified from canine liver. Substrate depletion was observed in the presence of recombinant CYP3A12, whereas depletion did not substantially occur when microsomes were pre‐incubated with polyclonal antibodies against CYP3A12. These findings confirmed that CYP3A12 is the major cytochrome P450 isoform responsible for the metabolism of vinblastine in dogs.