4个品种猕猴桃发酵酒品质评价

【目的】研究猕猴桃品种对猕猴桃果酒品质的影响,探索理化指标与感官评价的相关性,为通过原料筛选生产优质猕猴桃酒提供依据。【方法】以生理成熟期的金艳、翠香、翠玉和东红4个品种猕猴桃为原料,经过后熟软化、带皮破碎、添加果胶酶及ADT活性干酵母发酵等工序,酿制4个品种猕猴桃发酵酒,测定4种猕猴桃原料的理化指标（可溶性固形物（TSS）、总酸（TA）、固/酸比、pH值等）,分析4种发酵酒在理化参数（TSS、TA、酒精度、残糖、挥发酸、可溶性蛋白）、抗氧化物质（总酚、维生素C、黄酮）、香气物质（酯类、醇类、萜烯类等）和感官品质（外观、香气、口感、风格）等方面的差异,研究不同品种猕猴桃果酒间感官指标与理化指标的关系。【结果】4个品种猕猴桃酿酒原料的TSS、TA和pH,以及4种果酒的TSS、TA、酒精度、可溶性蛋白及总酚、Vc、黄酮等均表现出不同程度的差异,发酵酒的酒精度与后熟果实原料的pH值之间存在极显著负相关。在4种猕猴桃果酒中,共检测出7类83种挥发性化合物,其中共有类型6类,共有物质24种,以酯类化合物的种类最多、相对含量最大。4种猕猴桃酒香气成分总质量浓度由大到小分别为金艳（3 907.28 μg/L）＞东红（3 493.22 μg/L）＞翠玉（3 176.01 μg/L）＞翠香（2 768.17 μg/L）。4种猕猴桃酒的感官评分中,外观和总分存在一定差异,总分由高到低依次为东红＞金艳＞翠香＞翠玉。感官指标与理化指标的相关性分析表明,不同品种猕猴桃果酒的外观与其可溶性蛋白质量浓度呈显著负相关,香气与芳樟醇和萜烯类物质质量浓度呈显著正相关,口感与原料果实的固/酸比呈显著正相关,而与黄酮质量浓度呈显著负相关。【结论】4个猕猴桃品种中,东红猕猴桃果酒的外观、香气、口感及与感官测试相关的理化指标表现最优,最适于酿制猕猴桃酒。     

牦牛MYL3基因的克隆及组织表达分析

【目的】研究牦牛MYL3基因的结构和表达特征,探究其生物学功能及影响牦牛肌肉生长发育的机制。【方法】以美仁牦牛cDNA为模板,PCR扩增MYL3基因编码区序列(CDS),利用MEGA11软件构建系统进化树,利用生物信息学软件对其编码蛋白进行分析。通过实时荧光定量PCR(RT-qPCR)检测MYL3基因在心脏、肝脏、脾脏、肺脏（参照）、睾丸、肌肉和脂肪组织中的相对表达量。【结果】牦牛MYL3基因CDS区长600 bp,共编码199个氨基酸,存在1个碱基突变位点,位于第165位(A>T)。系统进化分析结果显示,牦牛与普通牛的亲缘关系最近,与单峰驼的亲缘关系最远。生物信息学分析结果显示,牦牛MYL3蛋白为不稳定的亲水性蛋白,无信号肽,不存在跨膜结构,主要存在于细胞质内,有8个磷酸化位点和2个N 糖基化位点,蛋白的二级结构以α-螺旋为主,主要与调控肌肉生长发育的相关蛋白相互作用。RT-qPCR结果表明,MYL3基因在心脏中的表达量最高,极显著高于其他组织;肌肉中次之,与除心脏外的其他组织存在极显著差异。【结论】MYL3基因可能与牦牛心脏发育有关,对肌肉的生长发育和肉质有一定影响。     

桂花OfWRKY120基因鉴定及盐胁迫响应功能研究

【目的】根据前期基因家族鉴定分析得到OfWRKY120,对该基因盐胁迫响应的功能进行分析,为桂花盐胁迫下的调控机制研究奠定基础。【方法】利用生物信息学方法对OfWRKY120保守结构域、系统进化进行分析,并采用qRT-PCR技术研究盐胁迫下OfWRKY120在桂花叶片中的相对表达量。构建OfWRKY120超表达载体,使用亚细胞定位、酵母自激活检测等探究其特性,将该基因瞬时转化本氏烟草后,对烟草进行盐处理,利用DAB、NBT染色和相关生理指标的测定,解析瞬时转化OfWRKY120对本氏烟草耐盐性的影响。【结果】OfWRKY120基因全长为1 422 bp,编码474个氨基酸,存在WRKY superfamily保守结构域。盐胁迫能够激活桂花叶片中OfWRKY120的表达,在72 h时相对表达量最高,与对照有显著差异。亚细胞定位结果表明,OfWRKY120定位于细胞核。酵母自激活结果显示OfWRKY120无自激活活性。盐胁迫后,OfWRKY120瞬时过表达植株的超氧阴离子(O^-₂·)和脯氨酸含量显著高于空载对照,且DAB和NBT染色表现出更深的颜色。本氏烟草中的qRT-PCR结果显示,OfWRKY120显著降低了NbCAT的表达,而显著提高了NbP5CS1、NbP5CS2和NbP5CR的表达。【结论】OfWRKY120过表达增加了盐胁迫下本氏烟草中活性氧（ROS）的含量,导致植物对盐胁迫的敏感性提高。     

生物结皮生长发育对黄绵土抗侵蚀性能的影响

[目的] 黄土高原退耕还林(草)工程驱动的生物结皮发育可显著抑制土壤侵蚀,为探明土壤抗侵蚀性能随生物结皮生长发育的变化特征及其响应机制。[方法] 设置藻结皮、藓结皮和自然演替结皮3个处理,培育176天,系统研究生物结皮不同发育阶段及其类型差异对黄绵土抗侵蚀性能的影响。[结果] (1)生物结皮盖度、厚度、生物量、叶绿素和粗糙度随生长时间均显著增加,培育初期藓结皮较高,培育末期自然演替结皮反而最高。(2)随生物结皮发育,土壤黏结力呈幂函数增强,较初期增加39.8%~60.3%;质量损失率呈指数函数减小,较初期降低45.6%~57.3%;饱和导水率变化趋势较为复杂,但培育末期均最低(0.08~0.12 mm/min)。(3)土壤黏结力随生物结皮盖度、厚度、叶绿素、生物量和表面粗糙度的增加呈幂函数增长,质量损失率随生物结皮盖度、厚度、叶绿素、生物量和表面粗糙度的增加呈指数函数下降。(4)土壤抗侵蚀综合指数(C_ser)可表示为盖度(Cov)、厚度(T)和生物量(B)的幂函数(C_ser=0.279 Cov^0.194 T^0.188 B^0.119, R²=0.73, p＜0.05)。[结论] 黄土高原生物结皮发育可显著提高黄绵土抗侵蚀性能,藓结皮效果最强。     

浙江不同稻区耳叶水苋对苄嘧磺隆的抗性比较

采用琼脂法测定了长江三角洲地区宁绍平原和杭嘉湖平原5个稻区(宁波、绍兴、杭州、嘉兴和湖州)稻田耳叶水苋Ammannia arenaria不同生物型对苄嘧磺隆的抗性水平。结果表明,所采集的88种耳叶水苋生物型中,有96.6%已对苄嘧磺隆产生了抗性,其中,低抗(3RI≤10)、中抗(10RI≤50)和高抗(RI>50)的生物型分别占总数的22.7%、53.4%和20.5%。宁波NB0143-01和绍兴SX077生物型对苄嘧磺隆的RI值分别高达124.4和120.4。5个稻区中,绍兴地区耳叶水苋的抗性水平最高,平均RI值为53.1;宁波次之,平均RI值为35.1;湖州、杭州和嘉兴地区的抗性水平相对较低,平均RI值分别为24.1、19.9和18.7。表明稻田耳叶水苋对苄嘧磺隆的抗性程度较高,且在宁绍平原和杭嘉湖平原稻区已普遍出现抗性。其中在宁绍平原苄嘧磺隆对耳叶水苋的选择压较大,长期连续用药可能是该地区抗性水平高于其他区域的重要原因。这是有关耳叶水苋对苄嘧磺隆抗性种群分布的首次报道。     

代谢酶抑制剂PX对除草剂防除稗草的增效作用

为解决除草剂防除困难的问题,采用室内盆栽试验测定了PX、PM和Mn²⁺三种代谢酶抑制剂对精喹禾灵、莠去津、烟嘧磺隆、硝磺草酮和烯禾啶防除稗草Echinochloa crus-galli的增效作用,测定不同用量代谢酶抑制剂PX对这5种除草剂的增效作用及其与莠去津混用后稗草体内谷胱甘肽转移酶（glutathione S-transferase,GST）和细胞色素 P450（cytochrome P450,CYP450）活性的变化。结果显示,在3种代谢酶抑制剂中,代谢酶抑制剂PX对精喹禾灵、莠去津、烟嘧磺隆、硝磺草酮和烯禾啶5种除草剂的增效作用最显著。不同用量代谢酶抑制剂PX对不同除草剂的增效作用不同,其中75 g (a.i.)/hm²代谢酶抑制剂PX对莠去津的增效最佳,株防除效果和鲜重防除效果分别增加了35.33个百分点和37.33个百分点。莠去津与代谢酶抑制剂PX混用后,稗草体内GST和CYP450活性均较对照显著降低。表明除草剂与代谢酶抑制剂PX混用能增加除草剂对稗草的防除效果,可以达到减施除草剂的目的。     

Effects of exosomes derived from hypoxia-preconditioned umbilical cord mesenchymal stem cells on function of endothelial cells

AIM To investigate the effect of exosomes derived from hypoxia-preconditioned human umbilical cord mesenchymal stem cells (hUCMSCs) on proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVECs). METHODS hUCMSCs and HUVECs were isolated, cultured and identified. Exosomes derived from hUCMSCs were extracted by ultracentrifugation. The morphological change of exosomes was observed under transmission electron microscope. The particle size and concentration of exosomes were detected by nanoparticle tracking analysis, and the surface specific marker proteins of exosomes were determined by Western blot. hUCMSCs were divided into normoxia group and hypoxia group. The viability of hUCMSCs was measured by CCK-8 assay. HUVECs were divided into control group, normoxic exosome group and hypoxic exosome group. The proliferation of HUVECs was detected by EdU assay. The migration ability was detected by cell scratch assay and Transwell experiment. Tube formation ability was evaluated by tube formation experiment. RESULTS Compared with normoxia group, hypoxia pretreatment enhanced the viability and exosome release of hUCMSCs. Compared with normoxic exosome group, hypoxic exosomes enhanced the proliferation, migration and tube formation of HUVECs. CONCLUSION Exosomes derived from hUCMSCs under hypoxia enhances the proliferation, migration and tube formation of HUVECs.     

北纬43°地区日光暖棚牛舍的设计及其环境评价

根据不同季节的太阳高度角随地理纬度变化这一原理,确定牛舍塑料暖棚面与地面的最佳夹角,使冬季大寒(最冷)这天正午时分,太阳辐射的入射角与塑料棚面垂直,以其充分利用太阳的辐射热能,提高牛舍内的温度。通过对地处北纬43°地区通辽市科尔沁区余粮堡镇吴金宝村设计的日光暖棚牛舍的温热环境和舍内空气质量的测定表明:大寒这天舍外气温-20￣-8℃下,牛舍内的温度平均5.25℃,有害气体浓度均低于中华人民共和国国家标准,达到了防寒抗灾的目的。     

花粉多糖注射剂临床前抗肿瘤药效学预试

花粉多糖对小鼠Lewis肺癌(足趾接种)模型以200mg/kg,100mg/kg,50mg/kg三个剂量iv×10qd方案给药,实验显示高、中、低三个剂量组抑瘤率分别为48.01%,43.09%,31.38%。免疫实验以上述相同的给药方案,花粉多糖对荷Lweis肺癌小鼠NK细胞活性具有明显的提高和促进作用。     

The 16MΔvjbR as an ideal live attenuated vaccine candidate for differentiation between Brucella vaccination and infection

Brucellosis brings great economic burdens for developing countries. Live attenuated vaccines are the most efficient means for prevention and control of animal Brucellosis. However, the difficulties of differentiating of infection from vaccine immunization, which is essential for eradication programs, limit their applications. Therefore, the development of a vaccine that could differentiate infection from immunization will overcome the limitations and get extensive application. VjbR is a quorum sensing regulator involving in Brucella's intracellular survival. The vjbR∷Tn5 mutants have been proven effective against wild type strain challenge, implying its possibility of use in vaccine candidate development. To further evaluate this candidate gene, in the present study, the antigenicity of purified recombinant VjbR protein was analyzed. Antibodies to Brucella melitensis VjbR could be detected in sera from patients and animals with brucellosis but not in control ones, implying the potential use of this protein as a diagnostic antigen. Then a vjbR mutant of B. melitensis 16M was constructed by replacing the vjbR with kanamycin gene. The mutant showed reduced survival in macrophage and mice. Vaccination of BALB/c mice with 16MΔvjbR conferred significant protective immunity against B. melitensis strain 16M challenges, being equivalent to which induced by the license vaccine Rev.1. The vjbR deletion mutant elicited an anti-Brucella-specific immunoglobulin G response and induced the secretion of gamma interferon and interleukin-10. The most importance is that, the use of vjbR mutants as vaccines in association with diagnostic tests based on the VjbR antigen would allow the serological differentiation between infected and vaccinated animals. These results suggest that 16MΔvjbR is an ideal live attenuated vaccine candidate against B. melitensis and deserves further evaluation for vaccine development.