整合素αv在牦牛繁殖周期输卵管不同部位中的表达与定位

旨在探究整合素αv在牦牛繁殖周期输卵管不同部位的表达差异,为进一步了解整合素αv在牦牛生殖过程中发挥的作用提供理论依据。本研究采集卵泡期、黄体期和妊娠期健康母牦牛（3~6岁）的输卵管伞部、壶腹部和峡部样品,将卵泡期、黄体期和妊娠期输卵管分别按照伞部、壶腹部和峡部分为9组,采用实时荧光定量PCR（qRT-PCR）和蛋白免疫印迹法（Western-blotting,WB）检测整合素αv基因和蛋白的表达情况,运用免疫组织化学法检测整合素αv的定位。结果表明：1）qRT-PCR结果显示,在卵泡期,整合素αv基因在输卵管伞部的表达量最高,极显著高于壶腹部和峡部（P<0.01）;在黄体期,整合素αv基因在输卵管峡部的表达量最高,极显著高于伞部和壶腹部（P<0.01）;在妊娠期,整合素αv基因在输卵管伞部的表达量最高,极显著高于壶腹部和峡部（P<0.01）。2）Western-blotting结果显示,在卵泡期,整合素αv蛋白在输卵管伞部的表达量最高,极显著高于壶腹部和峡部（P<0.01）;在黄体期,整合素αv蛋白在输卵管峡部的表达量最高,极显著高于伞部和壶腹部（P<0.01）;在妊娠期,整合素αv蛋白在输卵管峡部和伞部的表达差异不显著,但二者极显著高于壶腹部（P<0.01）。3）免疫组织化学结果显示,整合素αv主要分布于输卵管伞部、壶腹部和峡部的纤毛细胞、分泌细胞、基细胞、肌层和浆液腺中。结果显示,整合素αv在牦牛繁殖周期输卵管不同部位中的表达存在显著差异,说明其可能参与了牦牛受精及早期胚胎发育等一系列生殖过程,为牦牛的繁殖性能研究提供了基础资料。     

基于生命周期评价的相同食物当量玉米与紫花苜蓿生产环境影响比较研究

为了探讨种植业结构调整对于我国环境的影响,本研究运用生命周期评价方法,计算了甘肃省民勤县农户水平2014与2015年从农资生产到农户入仓范围生产1 kg玉米籽粒及1 kg紫花苜蓿鲜草的环境影响,并使用基于蛋白质和热量的计量单位—食物当量(FEU),比较分析了1个FEU玉米籽粒和紫花苜蓿生产的全生命周期环境影响差异。结果表明,生产1 kg玉米籽粒和1 kg紫花苜蓿鲜草全生命周期的一次性能源消耗(PED)分别为9.35和1.22 MJ,水资源消耗(WU)分别为889.33和144.37 kg,矿物和化石资源消耗(DAR)分别为0.13和0.02 kg antimony-eq,气候变化潜值(GWP)分别为1.21和0.10 kg CO₂-eq,可吸入无机物(RI)分别为4.23×10^-3和1.88×10^-4 kg PM_2.5-eq,光化学臭氧合成(POFP)分别为2.41×10^-3和1.71×10^-4 kg NMVOC-eq,环境酸化潜值(AP)分别为8.55×10^-3和8.03×10^-4 kg SO₂-eq,淡水富营养化(FEP)分别为1.20和0.09 kg P-eq,生态毒性(ecotoxicity)分别为1.26×10^-2和1.49×10^-3 CTU。1个FEU紫花苜蓿生产的PED、WU、DAR、GWP、RI、POFP、AP、FEP和ecotoxicity则分别为玉米籽粒的20.50%、25.43%、21.08%、12.99%、6.98%、11.15%、14.76%、12.31%和18.58%。因而考虑到苜蓿的食物-经济比较优势,目前应给予其不少于粮食作物的种植补贴。并且如果将我国的部分玉米种植改为苜蓿种植,则是最便捷、经济的既能满足我国食物结构需求,又能减少农业生产的资源消耗与环境污染的措施。本研究同时也为在我国深入开展粮改饲提供了一定的立论基础。     

生物地球化学模型DNDC的研究进展与碳动态模拟应用

脱氮脱碳模型(denitrification-decomposition,DNDC)是通过计算反硝化和有机质分解来模拟氮和碳从土壤丢失而转移入大气时的主要生物地球化学过程模型。作为目前国际上最成功的模拟陆地生物地球化学循环的模型之一,本文主要阐述了DNDC模型的发展过程及其结构,整理了DNDC模型在碳动态模拟过程中的主要研究进展,总结了当前DNDC模型在碳动态模拟领域的应用热点和优势。     

Evaluating environmental impacts of the Japanese beef cow–calf system by the life cycle assessment method

The objectives of this study were to evaluate the environmental impacts of a beef cow–calf system using a life cycle assessment (LCA) method and to investigate the effects of scenarios to reduce environmental impacts on the LCA results. The functional unit was defined as one marketed beef calf, and the processes associated with the cow–calf life cycle, such as feed production, feed transport, animal management, the biological activity of the animal and the treatment of cattle waste were included in the system boundary. The present results showed that the total contributions of one beef calf throughout its life cycle to global warming, acidification, eutrophication and energy consumption were 4550 kg of CO₂ equivalents, 40.1 kg of SO₂ equivalents, 7.0 kg of phosphate (PO₄) equivalents and 16.1 GJ, respectively. The contribution of each process to the total environmental impact in each environmental impact category showed a similar tendency to the contribution of each process in each environmental category reported in the case of the beef fattening system as a whole. The results from this analysis showed that shortening calving intervals by 1 month reduced environmental impacts by 5.7–5.8% in all the environmental impact categories examined in the current study, and increasing the number of calves per cow also reduced environmental impacts in all the categories, although the effects were smaller.     

Feline vaccine‐associated sarcomagenesis: Is there an inflammation‐independent role for aluminium?

Aluminium has been found in feline vaccine‐associated sarcomas. In this study, we investigated the potential for aluminium to contribute directly to tumourigenesis. Our results indicated that an aluminium hydroxide adjuvant preparation was cytotoxic and mutagenic in human‐Chinese hamster ovary (CHO) hybrid cells in vitro. Moreover, CHO cells deficient in DNA double strand break (DSB), but not single‐strand break (SSB), repair, were particularly sensitive to aluminium exposure compared with repair proficient cells, suggesting that aluminium is associated with DSBs. In contrast to CHO cells, primary feline skin fibroblasts were resistant to the cytotoxic effects of aluminium compounds and exposure to an aluminium chloride salt promoted cell growth and cell cycle progression at concentrations much less than those measured in particular feline rabies vaccines. These findings suggest that aluminium exposure may contribute, theoretically, to both initiation and promotion of tumours in the absence of an inflammatory response.     

紫花苜蓿种子吸胀期胚根线粒体AsA-GSH循环对低温胁迫的响应

为探究胚根线粒体抗坏血酸-谷胱甘肽(AsA-GSH)循环响应低温胁迫的抗氧化作用机制,以紫花苜蓿种子为材料,研究了不同温度(10和20℃)处理下发芽特性、不同吸胀时间(6、12和24 h)胚根线粒体AsA-GSH循环酶活性、抗氧化物以及过氧化氢(H₂O₂)含量的变化规律。结果表明,吸胀12和24 h后,10℃处理的胚根线粒体H₂O₂含量高于20℃。吸胀24 h期间,10℃处理的胚根线粒体谷胱甘肽还原酶(GR)活性均低于20℃。吸胀12和24 h后,10℃处理的胚根线粒体抗坏血酸(AsA)含量均低于20℃。10℃条件下吸胀24 h后,与20℃处理相比,胚根线粒体谷胱甘肽(GSH)含量显著降低(P<0.05)。苜蓿种子在10℃条件下吸胀萌发时,主要通过降低AsA-GSH循环中GR、单脱氢抗坏血酸还原酶(MDHAR)、过氧化物酶(POD)活性和AsA、GSH含量,使胚根线粒体内的H₂O₂积累,产生氧化损伤,继而影响种子萌发的正常进程。     

The second-generation curcumin analogue RL71 elicits G2/M cell cycle arrest and apoptosis in canine osteosarcoma cells

Canine osteosarcoma is an aggressive cancer, comprising 85% of canine bone neoplasms. Current treatment practices of surgery and chemotherapy increase 1-year survival by only 45%. The curcumin analogue RL71, has demonstrated potent in vitro and in vivo efficacy in several models of human breast cancer through increased apoptosis and cell cycle arrest. Thus, the present study aimed to investigate efficacy of curcumin analogues in two canine osteosarcoma cell lines. Osteosarcoma cell viability was assessed using the sulforhodamine B assay and mechanisms of action were determined by analysing the levels of cell cycle and apoptotic regulatory proteins via Western blotting. Further evidence was obtained using flow cytometry to detect cell cycle distribution and the number of apoptotic cells. RL71 was the most potent curcumin analogue with EC₅₀ values of 0.64 ± 0.04 and 0.38 ± 0.009 μM (n = 3) in D-17 (commercial) and Gracie canine osteosarcoma cells, respectively. RL71 significantly increased the ratio of cleaved-caspase 3 to pro-caspase 3 and the level of apoptotic cells at the 2× and 5× EC₅₀ concentration (p < 0.001, n = 3). Furthermore, at the same concentration, RL71 significantly increased the number of cells in the G2/M phase. In conclusion, RL71 has potent cytotoxic activity in canine osteosarcoma cells triggering G2/M arrest and apoptosis at concentrations achievable in vivo. Future research should further investigate molecular mechanisms for these changes in other canine osteosarcoma cell lines prior to in vivo investigation.     

猪子宫内膜肥大细胞消涨规律初探

雌性小型猪１５头，分为二月龄、间情期、民情前期、发情期、妊娠期等六组。颈动脉放血处死，取子宫组织，经甲苯胺改良法和免疫组化法染色，观察肥大细胞的动态变化。子宫内膜肥大细胞的数量随个体的生长而增加。在发情周期中肥大细胞的数量呈规律性的变化：在民情前期一时性地剧增，至发情期陡然降至显著低于间情期的水平，在妊娠期也维持低于间情期的水平。本实验的结果提示，子宫内膜肥大细胞的数量可能反映子宫局部的免疫水平。     

伊犁鹅产蛋前后各期卵巢转录谱的构建及卵泡发育相关基因的分析

【目的】构建产蛋前后各期伊犁鹅卵巢转录组文库,结合生物信息学分析揭示不同产蛋期伊犁鹅卵巢组织差异表达基因,并鉴定出影响鹅卵巢发育的关键基因,为伊犁鹅的繁殖调控提供理论参考。【方法】选择处于开产期（KL组）、产蛋期（CL组）及休产期（XL组）的伊犁鹅各4只,屠宰后取其卵巢组织,用于构建卵巢转录组文库。通过新基因挖掘、基因差异表达、基因注释以及蛋白互作网络分析筛选出与鹅卵泡发育相关的候选基因;随机挑选8个差异表达基因,应用实时荧光定量PCR验证其表达情况。【结果】伊犁鹅卵巢组织学结果表明,在开产期时,伊犁鹅卵巢表面有大量初级卵泡,而产蛋期卵巢则显示出卵泡的层级性,在休产期卵巢中可观察到卵泡出现向内凹陷、闭锁的现象。通过转录组测序（RNA-Seq）,从构建的12个伊犁鹅卵巢cDNA文库中获得有效读数57 811 186~85 328 377条,Q30值均>93.38%,每个样品所产的测序读数于鹅参考基因组上的比对率在82.79%~89.24%。在卵巢中注释的新基因共有1 112个,单核苷酸多态性（SNP）位点总数为1 642 273~2 425 069个;SNP和插入缺失（InDel）均主要注释于内含子区域;各时期的可变剪切类型主要集中为最后1个外显子可变剪切（TSS）及第1个外显子可变剪切（TTS）。在KL vs CL、XL vs CL及XL vs KL组中分别有337、1 136和525个差异表达基因,共有差异表达基因为α2A肾上腺素能受体（ADRA2A）、表皮蛋白（CP）、非转移性黑色素瘤糖蛋白B （GPNMB）及α-1-抗胰蛋白酶样（LOC106033756）。GO功能富集分析发现,差异表达基因主要富集在肽基酪氨酸磷酸化的正调控、细胞黏附及质膜外侧等过程。KEGG通路富集分析发现,差异表达基因主要显著富集于神经活性配体-受体相互作用、ECM-受体相互作用及类固醇生物合成等。结合蛋白互作网络分析,筛选到与鹅卵巢发育相关的潜在调控因子Bruton’s酪氨酸激酶（BTK）、血小板源性生长因子受体α（PDGFRA）、整合素β3（ITGB3）等。实时荧光定量PCR结果显示,RNA-Seq结果准确可靠。【结论】本研究揭示了产蛋前后各期伊犁鹅卵巢组织中的基因表达差异,筛选到影响伊犁鹅卵巢发育的神经活性配体-受体相互作用、ECM-受体相互作用、类固醇生物合成等重要通路与BTK、PDGFRA和ITGB3等关键候选基因,为了解鹅卵巢组织调控产蛋性能的分子机制提供了理论支撑。