藏绵羊KLF7基因表达特征分析及其过表达对前脂肪细胞增殖分化的影响

【目的】分析藏绵羊Krüppel样因子7(Krüppel-like factor 7,KLF7)基因表达特征,研究过表达该基因对前脂肪细胞增殖及分化的影响。【方法】从藏绵羊脂肪组织中分离前脂肪细胞进行培养及成脂诱导,应用实时荧光定量PCR技术检测KLF7基因在藏绵羊7个组织(大脑、皮下脂肪、肾脏、背最长肌、瘤胃、睾丸和回肠)和前脂肪细胞不同分化阶段(第0、2、4和8天)的mRNA相对表达水平;应用RT-PCR方法从藏绵羊脂肪组织中扩增KLF7基因CDS区序列,并将其连接到pcDNA3.1(+)真核表达载体获得pcDNA3.1-KLF7过表达质粒,转染前脂肪细胞;应用实时荧光定量PCR方法检测脂肪细胞增殖及分化标志基因mRNA表达水平;采用EdU和CCK-8方法分别检测过表达KLF7基因对EdU阳性细胞数和细胞活力的影响;采用油红O染色检测过表达KLF7基因后脂肪细胞脂滴生成量。【结果】KLF7基因在藏绵羊7个组织中均有表达,其中在大脑中的表达量最高,其次为皮下脂肪和肾脏,均显著高于其他组织(P<0.05);诱导分化第2、4和8天脂肪细胞mRNA表达量均显著高于分化前(P＜0.05),且分化第2天表达量最高;pcDNA3.1-KLF7过表达质粒转染前脂肪细胞2 d后显著或极显著抑制增殖标志基因CDK4、CyclinB1和CyclinD1的表达水平(P＜0.05;P＜0.01),极显著降低细胞活力及EdU阳性细胞数量(P＜0.01);pcDNA3.1-KLF7过表达质粒转染前脂肪细胞,诱导分化8 d后,脂肪细胞分化标志基因PPARγ、Glut4和ELOVL6的mRNA相对表达水平显著或极显著下调(P＜0.05;P＜0.01),且脂质沉积极显著减少(P＜0.01),表明过表达KLF7基因可抑制藏绵羊前脂肪细胞增殖及分化。【结论】KLF7基因在藏绵羊多个组织中广泛表达,且大脑、皮下脂肪、肾脏中表达量较高;诱导分化后脂肪细胞表达量显著高于分化前,且分化第2天表达量最高;过表达KLF7基因可抑制藏绵羊前脂肪细胞的增殖及分化。试验结果为阐明藏绵羊脂肪沉积的分子调控机制提供了基础数据。     

1,1—二甲基—7—异丙基—1,2,3,4—四氢萘—6—磺酸钠的合成

文章探讨了１，１－二甲基－７－异丙基－１，２，３，４－四氢萘－６－磺酸钠合成的磺化工艺。通过实验室的试验，确定了适宜的工艺条件，１，１－二甲基－７－惜内基－１，２，３，４，－四四氢萘与浓度硫酸的配比为１：１０．５（ｍｏｌ比），反应温度９０±２℃，反应时间２０ｍｉｎ，产品得率达８０．３％。同时测定了产物表面活性性能，可用作表面活性剂。     

Preparation of novel reagents 4-alkoxytrityl chlorides and their reaction with methyl Α-d-glucoside

A series of novel 4-O-alkoxytrityl chlorides (1) with different chain lengths was synthesized as a novel reagent for obtaining 6-O-alkylated cellulose with high regioselectivity via trityl groups in one reaction step without the use of any protective groups. These chlorides were reacted with methyl -d-glucoside, which was used as a model compound, to examine the reactivities toward the primary hydroxyl groups of cellulose to afford a series of 6-O-alkylated methyl -d-glucosides in high yields. The product compounds were found to have interesting solubilities and thermal properties. Thus, newly prepared trityl chloride derivatives were found to be useful regioselective derivatization reagents on the primary hydroxyl group in carbohydrates, especially in cellulose.     

某牛场粪源大肠杆菌0157∶H7的分离鉴定

为了解重庆市某牛场牛群大肠杆菌O157∶H7的感染情况,我们以牛场犊牛为研究对象,采集其新鲜粪便,经LB培养基增菌培养、免疫磁珠富集后,在山梨醇麦康凯琼脂上划线培养,最后PCR扩增eaeA基因目的片段,以此分离鉴定粪源大肠杆菌O157∶H7。结果显示：在所采集的39份犊牛粪便样品中,分离得到一株符合大肠杆菌O157∶H7生长特点且含有eaeA基因的菌株。     

陕西富硒与非富硒地区山羊组织硒含量和GPX-7基因表达的比较

为了探讨陕西紫阳县(富硒地区)山羊各组织中微量元素硒的含量和谷胱甘肽过氧化酶7(Glutathione Peroxidase 7,GPX-7)基因的表达情况,以及组织中的硒沉积量对GPX-7基因表达的影响。利用原子荧光光谱法对年龄相仿、体质量相近的紫阳县双安镇(n=7)和广成镇(n=7)和宝鸡市陈仓区白山羊体内硒含量进行了测定;同时运用实时荧光定量PCR(qRT-PCR)相对定量技术测定GPX-7基因mRNA的相对表达量。紫阳双安镇山羊硒含量显著高于紫阳广成镇(P0.05),同时紫阳县均显著高于宝鸡陈仓区(P0.05);但是紫阳双安镇与紫阳广成镇山羊肺脏中硒含量差异不显著(P0.05)。GPX-7基因在各组织器官中广泛表达,但是不同地区的山羊表现出不同的组织差异性。紫阳双安镇山羊心脏、脾脏、肾周脂肪和背最长肌组织中mRNA水平与紫阳广成镇差异不显著(P0.05),但是紫阳地区均显著高于宝鸡陈仓区(P0.05)。在肺脏、肾脏、股二头肌和淋巴组织中,来自不同地区的山羊GPX-7基因mRNA的表达量地差异均两两显著(P0.05),其中紫阳广成镇山羊肺脏的表达量显著高于其他两个地区(P0.05)。由此可见,山羊各组织器官中微量元素硒的沉积量和GPX-7基因mRNA的表达量表现出较为一致的地域趋势,即紫阳双安镇紫阳广成镇宝鸡陈仓区,这表明微量元素硒上调GPX-7基因的表达,但是具有组织差异性。     

The impact of weight loss on circulating cytokines in Beagle dogs

Chronic low-grade inflammation in obesity is characterized by an increased production of pro-inflammatory and chemotactic cytokines that are contributing to insulin resistance and related co-morbidities. Cytokines act in networks and exhibit pleiotropic effects so we investigated the circulating levels of a wide array of cytokines (pro and anti-inflammatory, chemotactic and growth factors) in a canine model of weight loss. The dogs served as their own control in order to study the impact of weight loss independent of potential confounding factors, such as history of excess weight or gender. While low-grade inflammation had been previously investigated in obese dogs by measuring changes in adipokines, acute phase proteins and key pro-inflammatory cytokines, to the best of our knowledge this is the first study to evaluate how weight loss impacts a wide array of circulating cytokines.Eighteen overweight Beagle dogs were recruited (six spayed females and 12 neutered males), and none of them were grossly obese according to the body condition score (BCS). All the dogs reached an ideal weight by the end of the program. Parameters were assessed before (baseline), at mid-point (month 3) and at end-point (month 6). Plasma GM-CSF, IL-2, Il-4, IL-6, IL-7, IL-8, IL-10, IL-15, IL-18, IFNγ, IP-10, TNFα, monocyte chemotactic protein 1 (MCP-1), keratinocyte chemokine (KC) were measured with canine multiplex immunoassays. Fat mass was assessed by dual energy X-ray absorption (DEXA).Several cytokines decreased throughout the weight loss program (p < 0.01) and were correlated with the percentage of fat measured by DEXA (p < 0.05): chemotactic (MCP-1), growth factors (GM-CSF, IL-7 and IL-2), and pro-inflammatory (KC and IL-18). We could not show trends for several cytokines, possibly because their level may be lower than the assay sensitivity: anti-inflammatory (IL-4 and IL-10), and pro-inflammatory (IL-6 and TNFα).In conclusion, while our findings for several pro-inflammatory and chemotactic cytokines are in accordance with human and rodent studies, we may have identified additional cytokines, such as growth factors, related to obesity-induced low-grade inflammation. Considering the weight loss was enabled by an adjusted diet, the role of this association of cytokines in insulin resistance and related co-morbidities needs to be clarified. Our results could help better understand the cytokine biology in dogs, and as such are relevant for further elucidating the relationship between immune function and metabolism/nutrition.     

microRNA let-7调控动物个体发育的研究进展

microRNA(miRNA)是一类广泛存在于多细胞动物中的进化保守的大小为18~25nt的非编码小分子RNA,可以通过与靶基因mRNA的非编码区(3′UTR)结合导致mRNA降解,或阻断mRNA翻译而调节基因表达。let-7是在线虫中发现的具有转录后调节功能的小分子RNA,具有高度的保守性,研究发现,let-7参与动物个体多个器官组织的发育过程。作者综述了近年来let-7参与调控脑、神经系统、心肺系统和肌肉发育等组织器官的研究成果,初步阐述了let-7调控组织器官发育的作用机制,以期为进一步探索let-7在动物体内的功能奠定基础。     

基于改进YOLO v7的轻量化樱桃番茄成熟度检测方法

为在自然环境下自动准确地检测樱桃番茄果实的成熟度,实现樱桃番茄果实自动化采摘,根据成熟期樱桃番茄果实表型特征的变化以及国家标准GH/T 1193—2021制定了5级樱桃番茄果实成熟度级别(绿熟期、转色期、初熟期、中熟期和完熟期),并针对樱桃番茄相邻成熟度特征差异不明显以及果实之间相互遮挡问题,提出一种改进的轻量化YOLO v7模型的樱桃番茄果实成熟度检测方法。该方法将MobileNetV3引入YOLO v7模型中作为骨干特征提取网络,以减少网络的参数量,同时在特征融合网络中加入全局注意力机制(Global attention mechanism, GAM)模块以提高网络的特征表达能力。试验结果表明,改进的YOLO v7模型在测试集下的精确率、召回率和平均精度均值分别为98.6%、98.1%和98.2%,单幅图像平均检测时间为82 ms,模型内存占用量为66.5 MB。对比Faster R-CNN、YOLO v3、YOLO v5s和YOLO v7模型,平均精度均值分别提升18.7、0.2、0.3、0.1个百分点,模型内存占用量也最少。研究表明改进的YOLO v7模型能够为樱桃番茄果实的自...     

基于改进YOLOv3网络模型的茶草位置检测算法

精准高效的茶草识别是智能茶园植保机械进行除草工作的关键。针对目前茶园除草智能化程度较低等问题,提出改进YOLOv3网络模型的茶草检测算法。首先,分季节和时间段,在多个茶叶品种的种植园中以自适应的距离和角度采集茶草混合图像并建立试验数据集。接着,使用K均值聚类算法重新设计先验锚框尺度。然后,以YOLOv3网络模型为基础,选取17×17的网格划分图像区域;采用残差网络（ResNet）作为主干网;加入过程提取层,增强草株检测性能。最后在原损失函数中引入广义交并比损失。通过消融试验和不同目标检测算法对比试验验证此改进算法对茶树与杂草的检测效果。试验结果表明,改进 YOLOv3网络模型对杂草的检测精确率和召回率分别为85.34%和91.38%,对茶树的检测精确率和召回率最高达到82.56%和90.12%;与原YOLOv3网络模型相比,检测精确率提高8.05%,并且每秒传输帧数达到52.83 Hz,是Faster R-CNN网络模型的16倍。这些数据说明所提算法在茶园复杂环境下,不仅对于茶树和杂草具有更好的识别效果,而且满足实时检测的要求,可以为智能茶园植保机械提供技术支持。