基于岗位对接和任务驱动的高职课程“食品微生物检测技术”的改革实践

基于职业岗位和典型工作任务进行高职课程"食品微生物检测技术"改革探索,分别对课程的教学目标、教学内容、教学实施和教学评价方法进行改革,教学实践证明此课改模式能有效激发学生学习兴趣,提高学生综合素质,培养的学生能适应工作岗位需求,取得了较好的教学效果。     

增温和刈割对高寒草甸土壤呼吸及其组分的影响

评估土壤呼吸及其组分对增温等全球变化的响应对于预测陆地生态系统碳循环至关重要。本研究利用红外线辐射加热器(Infrared heater)装置在青藏高原高寒草甸生态系统设置增温和刈割野外控制实验。通过测定2018年生长季(5—9月)土壤呼吸和异养呼吸,探究增温和刈割对土壤呼吸及其组分的影响。研究结果表明：(1) 单独增温使土壤呼吸显著增加31.65% (P<0.05),异养呼吸显著增加27.12% (P<0.05),土壤自养呼吸没有显著改变(P>0.05);单独刈割对土壤呼吸和自养呼吸没有显著影响(P>0.05),单独刈割刺激异养呼吸增加32.54% (P<0.05);(2) 增温和刈割之间的交互作用对土壤呼吸和异养呼吸没有显著影响(P>0.05),但是对自养呼吸的影响是显著的(P<0.05),土壤呼吸和异养呼吸的季节效应显著(P<0.05);(3)土壤呼吸及其组分与土壤温度均成显著指数关系,与土壤湿度呈显著的正相关关系(P<0.05),处理影响它们的响应敏感性。本研究表明青藏高原东缘高寒草甸土壤碳排放与气候变暖存在正反馈。     

补料工艺对两株法夫酵母菌株产虾青素的影响

【目的】考察不同补料工艺对法夫酵母菌株生长和虾青素合成的影响。【方法】对法夫酵母JMU-VDL668和JMU-MVP14菌株在7 L罐中进行分批及分批补料培养; 同时, 测定发酵过程中生物量、虾青素和葡萄糖含量的变化。【结果】采用恒DO补料, 法夫酵母JMU-VDL668菌株获得的生物量最大(64.6 g/L), 是分批培养的2.2倍; 采用恒pH补料发酵, 虾青素的产量最高(20.6 mg/L), 是分批培养的1.5倍。与JMU-VDL668菌株不同, 虾青素高产菌株JMU-MVP14菌株采用恒pH补料, 获得生物量最大(48.5 g/L), 但虾青素产量大大降低(仅17.5 mg/L); 采用脉冲补料, 虾青素产量最高, 达到414.1 mg/L, 与分批发酵相比提高了200.2%; 采用恒DO补料, 生物量(38.5 g/L)和虾青素产量(403.2?mg/L)增加显著, 与分批发酵相比分别提高了133.1%和192.3%。【结论】不同补料工艺对法夫酵母菌株生产虾青素影响很大。其中, 采用恒pH补料工艺, 法夫酵母JMU-VDL668菌株可以获得最高的虾青素产量, 而采用脉冲补料工艺, 最适于法夫酵母JMU-MVP14菌株发酵生产虾青素。     

禽源大肠杆菌耶尔森氏菌强毒力岛核心基因的检测及其irp2和int基因同源性

【目的】为了提高禽源大肠杆菌中耶尔森氏菌强毒力岛(HPI)的检测效率, 了解高分子量铁调节蛋白2基因(irp2)和整合酶基因(int)在不同株禽源HPI+大肠杆菌间的同源性, 进一步揭示禽源大肠杆菌HPI的转移规律。【方法】利用L16(44)正交试验设计, 建立针对HPI核心基因irp2和fyuA的双重PCR, 运用双重PCR方法检测禽源大肠杆菌临床分离株, 并对检出的7株HPI阳性(HPI+)大肠杆菌进行irp2和int基因测序及同源性分析, 同时结合这7株大肠杆菌的ERIC-PCR分析结果, 对比分析int基因的分布特点。【结果】结果显示, 新建立的双重PCR能特异性扩增出HPI核心基因; ERIC-PCR分析显示, HPI+大肠杆菌间差异均大于5%; HPI+大肠杆菌irp2基因高度保守(同源性大于99%), 而int基因虽然都位于asn-tRNA位点, 但基因序列在部分菌株间存在较大差异。【结论】建立了一种可以用于HPI的流行病学调查和实验室诊断的双重PCR方法, 并推测区域外同源重组可能是HPI基因在大肠杆菌间水平转移的主要方式。     

干旱区不同地理种群骆驼刺元素组成及表面结构特征的对比研究

碳(C)、氮(N)、磷(P)元素在植物各器官中的组成,及植物表面结构差异是其对外界环境适应性的重要表征。以干旱区3个地理种群骆驼刺(塔里木盆地策勒种群,吐鄯托盆地托克逊种群,准噶尔盆地阜康种群)为研究对象,通过对植株各器官C,N,P元素组成的测定及表面形貌观测,对其在不同环境中的适应特性进行了比较研究。结果表明:(1)同一种群的骆驼刺,C元素在各器官中的分配没有显著规律;N元素在叶片中含量最高,茎中最低;策勒种群和阜康种群的P含量,叶片中最高,茎与刺中差异不显著;但托克逊种群茎中P含量显著高于其他部位。(2)3个地理种群的骆驼刺叶中元素组成相比,策勒种群C,N含量均最高,托克逊种群C,N含量最低;3个种群叶片的P含量,C∶N,C∶P,N∶P值均没有显著差异。刺中元素含量相比,C含量没有显著差异;N含量阜康种群策勒种群托克逊种群。茎中元素含量相比,N含量差异不显著,但托克逊种群茎中P含量为其他两个种群的2倍,可能与托克逊土壤中高浓度的全N,速效N,速效P有关。(3)托克逊种群表皮极厚,蜡质非常致密,各器官气孔密度均高于其他两个种群;策勒种群比阜康种群叶表皮增厚,蜡质致密,但叶片气孔密度却减小;策勒种群和阜康种群茎与刺的气孔密度差异不显著。研究表明,策勒种群骆驼刺最适应当地环境条件:其叶片具有最高的C,N含量,且表面结构没有明显的干旱胁迫特征。托克逊种群表现出明显的干旱适应特征:其表皮增厚,蜡质致密,气孔密度增大,叶片C,N含量最低。虽然托克逊种群茎中P含量显著高于其他地理种群,但3个地理种群骆驼刺叶片中C∶N,C∶P,N∶P比值保持恒定,C∶N=30.6±4.3,C∶P=357.4±49.9,N∶P=12.0±2.4,说明骆驼刺能够保持较高内稳性,这也可能是其在新疆各地广泛生存的重要原因。     

链霉菌磷酸泛酰巯基乙胺基转移酶对载体蛋白的底物选择性的研究进展

磷酸泛酰巯基乙胺基转移酶(PPTase)催化脂肪酸合酶(FAS)、聚酮合酶(PKS)和非核糖体肽合成酶(NRPS)中载体蛋白从脱辅基形态转化为全辅基形态,对脂肪酸、PKS产物和NRPS产物的生物合成起着不可或缺的作用。本文介绍并总结了链霉菌PPTase对载体蛋白底物选择性的最新研究进展:Ⅲ型PPTase特异性催化同一个多肽链中ACP的辅基化;Ⅱ型PPTase倾向于催化Ⅰ型PKS中ACP和NRPS中PCP的辅基化;Ⅰ型PPTase倾向于催化Ⅱ型PKS中ACP和Ⅱ型FAS中ACP的辅基化;编码基因位于基因簇内的Ⅰ型/Ⅱ型PPTase倾向于催化编码基因位于同基因簇内的PKS/NRPS中ACP/PCP的辅基化;这些研究结果为阐明并改造链霉菌辅基化网络以提高特定次级代谢产物的产量提供了参考和借鉴。     

典型城市单元的土壤重金属溯源方法与实证研究

随着城市化进程的不断深入,土壤中重金属污染现状及其治理情况越来越受到重视,而查明污染源是有效治理污染的前提。源解析技术目前已广泛的应用于环境受体重金属来源解析实践中,总结了近年来土壤重金属成因分析的常用方法及原理,并提出了一种将多种方法相配合使用的方法体系。选取珠三角某市城郊农田作为研究对象,结果表明,Cd、Pb、Cu、Zn、As存在含量超过国家农用地筛选值的情况,其中Cd超标率高达60.1%。农业活动、工业生产、交通源和自然母质均对研究区土壤重金属的累积产生一定的贡献。正定矩阵因子分析法(PMF,Positive Matrix Factorization)模型模拟的Cd、Ni、Zn和Hg预测值与实测值线性拟合r~2均大于90%,其余元素r~2均大于60%,呈现出很好的相关性,满足研究需要。PMF模型和铅同位素比值法计算得到的交通及农业对土壤Pb累积的贡献率之和分别为86.0%和84.8%,PMF模型和物质流分析法计算得到的农业对土壤Cd的贡献率分别为86.7%和79.7%,结果均比较接近。结果表明正定矩阵因子法、同位素比值分析法,物质流分析法能很好的联用应用于土壤重金属源解析研究。     

Oxidized high-density lipoprotein promotes CD36 palmitoylation and increases lipid uptake in macrophages

Oxidized high-density lipoprotein (oxHDL) reduces the ability of cells to mediate reverse cholesterol transport and also shows atherogenic properties. Palmitoylation of cluster of differentiation 36 (CD36), an important receptor mediating lipoprotein uptake, is required for fatty acid endocytosis. However, the relationship between oxHDL and CD36 has not been described in mechanistic detail. Here, we demonstrate using acyl-biotin exchange analysis that oxHDL activates CD36 by increasing CD36 palmitoylation, which promotes efficient uptake in macrophages. This modification increased CD36 incorporation into plasma lipid rafts and activated downstream signaling mediators, such as Lyn, Fyn, and c-Jun N-terminal kinase, which elicited enhanced oxHDL uptake and foam cell formation. Furthermore, blocking CD36 palmitoylation with the pharmacological inhibitor 2-bromopalmitate decreased cell surface translocation and lowered oxHDL uptake in oxHDL-treated macrophages. We verified these results by transfecting oxHDL-induced macrophages with vectors expressing wildtype or mutant CD36 (mCD36) in which the cytoplasmic palmitoylated cysteine residues were replaced. We show that cells containing mCD36 exhibited less palmitoylated CD36, disrupted plasma membrane trafficking, and reduced protein stability. Moreover, in ApoE^−/−CD36^−/− mice, lipid accumulation at the aortic root in mice receiving the mCD36 vector was decreased, suggesting that CD36 palmitoylation is responsible for lipid uptake in vivo. Finally, our data indicated that palmitoylation of CD36 was dependent on DHHC6 (Asp-His-His-Cys) acyltransferase and its cofactor selenoprotein K, which increased the CD36/caveolin-1 interaction and membrane targeting in cells exposed to oxHDL. Altogether, our study uncovers a causal link between oxHDL and CD36 palmitoylation and provides insight into foam cell formation and atherogenesis.     

TBK1 Facilitates GLUT1-Dependent Glucose Consumption by suppressing mTORC1 Signaling in Colorectal Cancer Progression

Intestinal inflammation is a vital precipitating factor of colorectal cancer (CRC), but the underlying mechanisms are still elusive. TANK-binding kinase 1 (TBK1) is a core enzyme downstream of several inflammatory signals. Recent studies brought the impacts of TBK1 in malignant disease to the forefront, we found aberrant TBK1 expression in CRC is correlated with CRC progression. TBK1 inhibition impaired CRC cell proliferation, migration, drug resistance and tumor growth. Bioinformatic analysis and experiments in vitro showed overexpressed TBK1 inhibited mTORC1 signaling activation in CRC along with elevated GLUT1 expression without inducing GLUT1 translation. TBK1 mediated mTORC1 inhibition induces intracellular autophagy, which in turn decreasing GLUT1 degradation. As a rescue, blocking of autophagosome and retromer respectively via autophagy-related gene 7 (ATG7) or TBC1 Domain Family Member 5 (TBC1D5) silence diminished the regulation of TBK1 to GLUT1. GLUT1 staining presented that TBK1 facilitated GLUT1 membrane translocation which subsequently enhanced glucose consumption. Inhibitor of TBK1 also decreased GLUT1 expression which potentiated drug-sensitivity of CRC cell. Collectively, TBK1 facilitates glucose consumption for supporting CRC progression via initiating mTORC1 inhibition induced autophagy which decreases GLUT1 degradation and increases GLUT1 membrane location. The adaptive signaling cascade between TBK1 and GLUT1 proposes a new strategy for CRC therapy.