蛙皮素样肽家族及其受体氨基酸序列信息学比较分析

【目的】研究反刍动物蛙皮素样肽家族多肽及其受体的保守性。为不同动物,特别是反刍动物这2种蛙皮素样多肽及其受体蛋白抗原设计和活性多肽筛选等研究提供参考。【方法】采用生物信息分析学的方法,通过UniProt数据库比较牛的蛙皮素样肽家族胃泌素释放肽(Gastrin-releasing peptide, GRP)和神经介素B(Neuromedin B, NMB)2种多肽及其特异性受体GRP-R和NMB-R与其他动物氨基酸序列的差异性。【结果】13种动物成熟GRP多肽C-端的8个氨基酸序列完全相同,牛GRP氨基酸序列与绵羊、猪和豚鼠最高,分别为88.7%、77.8%和77.8%,与其他动物相似性低于74.1%。10种动物成熟NMB多肽C-端10个氨基酸序列完全相同。牛GRP-R与狗、马和猪等相似性最高,分别为96.1%、94.3%和94.0%,牛NMB-R与猪、人和马相似性最高,分别为92.3%、91.5%和91.0%;牛GRP-R与NMB-R相似性仅为62.2%。【结论】GRP和NMB的C-端氨基酸序列在不同动物之间均具有高度的保守性,而N-端为变化区域,且GRP-R和NMB-R氨基酸序列在动物之间保守性较高。     

Periplaneta americana peptide decreases apoptosis of pig-ovary granulosa cells induced by H2O2 through FoxO1

Oxidative stress can induce apoptosis of granulosa cells and lead to follicular atresia, thereby reducing the number of pigs giving birth. The aim of this study was to investigate the protective effect of Periplaneta americana peptide (PAP) on the apoptosis of the granulosa cells of pig ovaries (PGCs) induced by hydrogen peroxide (H₂O₂) via FoxO1. PGCs were treated with H₂O₂ to establish a cell apoptosis model. Cell viability was measured using the cell counting kit-8 (CCK-8) assay, and cell apoptosis was detected using flow cytometry. The malondialdehyde (MDA) level and nitric oxide (NO) content were detected to reflect the oxidative stress. Western blotting, qRT-PCR and overexpression were undertaken to determine the expression of FoxO1 and caspase-3, and immunofluorescence was used to detect FoxO1 in the nucleus and cytoplasm. PGCs were treated with 100 μM H₂O₂ for 6 hr, which resulted in oxidative damage and apoptosis and an apoptosis rate for PGCs of 32.95%. Next, PGCs were treated with 400 μg/ml PAP for 24 hr to repair the apoptosis induced by H₂O₂. PAP improved cell viability in H₂O₂-stimulated PGCs, the increased MDA level and NO content caused by H₂O₂ stimulation were reversed and the apoptotic rate of PGCs was reduced. The qRT-PCR and Western blotting results indicated that PAP decreased the H₂O₂-induced apoptosis and the expression of FoxO1 and caspase-3 in PGCs. The effect of PAP was the same following FoxO1 overexpression. FoxO1 was expressed in the nucleus when stimulated by H₂O₂ or overexpression; however, it migrated to the cytoplasm following PAP treatment. PAP decreased the apoptosis of PGCs induced by H₂O₂ by regulating FoxO1 expression and nuclear translocation.     

猪链球菌蛋白表面展示系统的构建及展示蛋白的初步观察

为构建猪链球菌(Streptococcus suis)蛋白表面展示系统,本研究通过序列分析,确定猪链球菌的LPxTG蛋白及其信号肽(SP)和胞壁锚定基序(CWA),通过PCR扩增Peno-SP、GFP、CWA的DNA片段并融合,构建强启动子Peno控制表达编码SP-GFP-CWA融合蛋白的DNA片段,将该重组DNA片段连接pSET2载体,获得蛋白表面展示质粒,转化猪链球菌,构建得到以GFP为报告蛋白的猪链球菌蛋白表面展示系统。结果显示,利用猪链球菌的10个LPxTG蛋白及其SP和CWA序列,构建了10个含有Peno-SP-GFP-CWA融合片段的重组pSET2表面蛋白展示质粒pSsPSD1至pSsPSD10,分别转化猪链球菌05ZYH33,PCR鉴定显示其中7个转化猪链球菌。采用western blot初步检测其展示蛋白,结果显示,7个转化阳性菌株均能有效表达GFP蛋白,以成熟GFP条带为指标,均表现出了一定的外源GFP表面展示水平,分别命名为SsPSD1、SsPSD2、SsPSD4、SsPSD7-SsPSD10,其中SsPSD1、SsPSD4、SsPSD8和SsPSD9表面展示水平相对较好,在猪链球菌表面展示外源蛋白方面具有很好的潜力。本研究首次尝试建立猪链球菌蛋白表面展示系统,为猪链球菌表面递呈外源蛋白或抗原提供了新的策略。     

日粮大豆活性肽对仔猪生长和相关理化指标及抗氧化性能的影响

为了探讨大豆活性肽对仔猪生长性能、血液指标及氧化应激参数的影响,试验选取仔猪48头,随机分为2组,每组设3个重复,每重复8头。对照组饲喂基础日粮,试验组饲喂用大豆活性肽替代2%豆粕的日粮。结果:添加2%大豆活性肽,20 d时,显著提高了仔猪粗脂肪、粗蛋白的消化率(P<0.05);血清总蛋白(TP)、血清白蛋白(ALB)和碱性磷酸酶(ALP)水平略高于对照组;50 d时,大豆活性肽显著提高了仔猪干物质、粗灰分、粗脂肪、粗蛋白、粗纤维的消化率(P<0.05),而且总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)活性及TP水平显著高于对照组(P<0.05);全程试验组日增重均显著的高于对照组,料重比、腹泻率显著低于对照组(P<0.05),然而对尿素氮(BUN)和甘油三脂(TG)影响不明显。     

城区道路景观共生模式研究——以金寨县将军大道为例

对小城镇沿河主干道的道路周边用地和道路中央分隔带进行景观设计和植物配置,将生态学中的共生机制原理运用到景观规划中,将地形、地貌、地物的自然景观,满足人类精神需要的人文景观,道路行车安全的功能景观和谐优化,初步探讨出一种共生机制下的小城镇道路景观共生模式。     

兴仁小白壳薏苡不同播期对产量的影响

为探索适宜兴仁县薏苡种植的最佳播种时期，在兴仁县开展了薏苡不同播期对产量影响的试验。试验共设9个播期，梯度为7 d，播种时间从3月19日－5月14日。试验结果表明，兴仁小白壳薏苡的最佳播期是3月19日－4月16日，并且在此期间播种，薏苡产量较高。     

我国中小银行成长性综合评价研究

通过构建商业银行成长性评价体系，利用突变级数法对14家中小银行2007~2010年的成长性进行评价，得出中小银行具有振荡向上的成长趋势，且股份制银行整体成长性优于城市商业银行，但对于影响成长性的各因素，银行间的表现不尽相同。这表明中小银行还需调整经营管理思路来保证成长的稳健和持续。     

Comparison of N-terminal pro-atrial natriuretic peptide and three cardiac biomarkers for discriminatory ability of clinical stage in dogs with myxomatous mitral valve disease

Plasma N-terminal pro-atrial natriuretic peptide (NT-proANP) concentration increases with progression of myxomatous mitral valve disease (MMVD) in dogs. This multicentre, prospective study compared plasma NT-proANP, N-terminal pro-brain natriuretic peptide (NT-proBNP), ANP, and cardiac troponin I (cTnI) concentrations in dogs with MMVD for their characteristics and discriminatory ability to detect cardiac dilatation and congestive heart failure (CHF). Thirty-six healthy dogs and 69 dogs with MMVD were included. Clinical variables were obtained via physical examination, thoracic radiography, and echocardiography. The discriminatory ability of each cardiac biomarker (CB) to determine the presence or absence of cardiac dilatation (event 1) and CHF (event 2) was evaluated using the receiver operating characteristic curves. Plasma NT-proANP, NT-proBNP, and ANP concentrations showed a significant association with the left atrium/aorta ratio (P<0.01). The area under the curve of plasma NT-proANP and NT-proBNP concentrations were 0.72 and 0.75, respectively in event1 and 0.72 and 0.76, respectively in event2. Plasma NT-proANP and NT-proBNP concentrations showed sensitivity 80.0 and 80.0%; specificity 67.6 and 64.7% in event1 (cutoff value; 8,497.81 pg/ml and 1,453.00 pmol/l, respectively) and sensitivity 85.7 and 81.0%; specificity 60.4 and 64.6% in event2 (cutoff value; 8,684.33 pg/ml and 1,772.00 pmol/l, respectively). In dogs with MMVD, plasma NT-proANP, NT-proBNP, and ANP concentrations increase with left atrial enlargement. Particularly, plasma NT-proANP and NT-proBNP concentrations appeared to be equally useful in the discriminatory ability to detect cardiac dilatation and CHF.     

抗菌肽粗提物对肉鸡生长性能、粪便微生物菌群结构及经济效益的影响

为分析抗菌肽粗提物对肉鸡生长性能、粪便微生物菌群结构及经济效益的影响,试验选择体重相近的21日龄白羽肉鸡600只,随机分成4组,每组15个重复,每个重复10只,1组为空白对照组饲喂基础日粮,试验2、3、4组在基础日粮中分别添加1.0%、1.5%、2.0%的抗菌肽粗提物,试验期为21 d。结果表明：（1）试验2、3、4组的平均采食均高于1组（P > 0.05）|试验3、4组的平均日增重较1组比分别提高11.0%、9.4%（P ＜ 0.05）,料重比较1组比分别降低18.5%、15.2% （P ＜ 0.05）|（2）试验3、4组粪便中大肠杆菌的含量较1组分别降低14.3%、18.0%（P ＜ 0.05）,试验3、4组粪便中乳酸杆菌、双歧杆菌的含量较1组比分别提高4.6%、6.7%、14.5%、12.3%（P ＜ 0.05）|（3）试验3、4组每只肉鸡经济效益较1组比分别提高19.3%、17.7%。综上,1.5%抗菌肽粗提物添加可以提高白羽肉鸡生长性能,改善粪便微生物菌群结构,提高其经济效益。 [关键词] 抗菌肽粗提物|白羽肉鸡|生长性能|粪便微生物菌群结构|经济效益