多能干细胞诱导分化为肾脏类器官的研究进展与挑战

用于分化为多种类型细胞的多能干细胞(PSC)体外培养技术已被广泛应用于生物学领域中.由PSC分化而来的肾脏类器官可基本还原生物体内肾脏的组织结构和部分功能,在肾脏疾病模型研究和药物筛选中有重要作用,继续改善肾脏类器官的结构、功能和成熟度将会对肾脏再生治疗提供极大的帮助.研究肾脏类器官的重点在于体外准确模拟体内肾脏的发育...     

Identifying suitable reference genes for developing and injured mouse CNS tissues

Accurate quantification of gene expression is fundamental for understanding the molecular, genetic and functional bases of tissue development and diseases. Quantitative real‐time PCR (qPCR) is now the most widely used method of quantifying gene expression due to its simplicity, specificity, sensitivity, and wide quantification range. The use of appropriate reference genes to ensure accurate normalization is crucial for the correct quantification of gene expression from the early development, maturation, aging to injury processes in the central nervous system (CNS). In this study, we have determined the expression profiles of 12 candidate housekeeping genes (ACTB, CYC1, HMBS, GAPDH, HPRT1, RPL13A, YWHAZ, PPIA, RPLP0, TFRC, GUS, and 18S rRNA) in developing mouse brain and spinal cord. Throughout development, there was a significant degree of fluctuations in their expression levels, indicating the importance and complexity of finding appropriate reference genes. Three software including BestKeeper, geNorm and NormFinder were used to evaluate the stability of potential reference genes. GUS was the most stable gene and GUS/YWHAZ were the most stable reference gene pair across different developmental stages in different CNS regions, whereas HPRT1 and GAPDH were the most variable genes and thus inappropriate to use as reference genes. Therefore, our results identified GUS and YWHAZ as the best combination of two reference genes for expression data normalization in CNS developmental studies. © 2017 Wiley Periodicals, Inc. Develop Neurobiol 78: 39–50, 2018     

不吸水链霉菌梧州新亚种胆固醇氧化酶基因的克隆与序列分析

克隆不吸水链霉菌梧州新亚种胆固醇氧化酶基因.以不吸水链霉菌梧州新亚种基因组DNA作为模板,根据已知的胆固醇氧化酶基因的保守序列设计简并引物进行PCR扩增,PCR产物与载体连接、转化,构建重组质粒.获得了不吸水链霉菌梧州新亚种胆固醇氧化酶基因重组质粒,序列测定并分析.     

ILK在肺鳞状细胞癌和肺腺癌的表达及其与临床病理特征和预后的关系

目的 探讨ILK在肺鳞状细胞癌和肺腺癌组织中的表达情况,及其与病理分型、肿瘤分化、分期、淋巴结转移及预后的关系。方法采用S-P免疫组织化学方法和Western Blot法,检测肺鳞状细胞癌和肺腺癌组织及相应癌旁肺组织中整合连接激酶（integrin-linkedkinase,ILK）的表达情况,并结合临床和病理资料进行分析。结果免疫组化结果显示：ILK在53／76（70％）的肺癌组织中阳性表达。其中鳞状细胞癌阳性率75％（33／44）,腺癌阳性率62．5％（20／32）,其表达与肺鳞状细胞癌的分化呈负相关（P〈0．01）,与临床分期（P〈0．01）、淋巴结转移（P〈0．01）呈正相关;与肺腺癌的临床分期（P〈0．01）和淋巴结转移（P〈0．01）正相关,与分化程度无相关性（P〉0．05）。同时,其表达与患者的生存时间呈负相关（P〈0．01）,与年龄、性别、肿瘤大小和组织类型等因素无关。Western Blot法进一步证实ILK在肺癌组织中的表达显著高于癌旁正常肺组织（P〈0．01）,其表达与肺癌的分化（P〈0．01）显著负相关。结论肺鳞状细胞癌和肺腺癌中,ILK与肺癌的侵袭和转移有关。ILK可作为判断肺鳞状细胞癌和肺腺癌预后的参考指标。     

Effects of light and plant growth regulators on the biosynthesis of vindoline and other indole alkaloids in Catharanthus roseus callus cultures

A callus strain with stable ability for vindoline synthesis was selected from many prepared Catharanthus roseus leaf calli to study the regulation of vindoline biosynthesis as well as other indole alkaloids. It was shown that light and plant growth regulators significantly influenced the biosynthesis of vindoline and other alkaloids as well as acidic and basic peroxidase activities. Light promoted vindoline and serpentine biosynthesis, and stimulated plastid development and peroxidase activity. However, 2,4-D suppressed the biosynthesis of all indole alkaloids and peroxidase activity. Our results suggest that light or plant hormones regulate vindoline, serpentine and other alkaloid biosynthesis and accumulation by influencing peroxidase activity and the differentiation status of callus cultures, especially chloroplast development. Some possible relationships between serpentine or vindoline biosynthesis and peroxidase activity are proposed.     

两个对浓核病毒(镇江株)具感性差异的家蚕品种的血液和围食膜蛋白的比较

家蚕品种间对浓核病毒(镇江株)具有不同的感染性,为了进一步探明家蚕对浓核病毒(镇江株)感性差异的分子机制,本文运用蛋白质电泳和质谱技术比较分析了两个对家蚕浓核病毒(镇江株)感性和抗性的近等基因系家蚕品种JS和NIL的血液和围食膜组织蛋白.结果表明:这两个家蚕品种的血液和围食膜组织蛋白组成差异很小.在血液组织蛋白中发现5个差异蛋白点,其中家蚕品种JS血液中有两个差异蛋白,可能分别为酪蛋白激酶和新型组织蛋白;在NIL血液组织中发现3个差异蛋白点,其中两个可能分别为线粒体延伸因子和类丝氨酸蛋白酶,另外1个的含量极显著高于JS,为血淋巴蛋白.在围食膜蛋白中共发现4个差异蛋白点,其中JS围食膜中有1个差异蛋白可能为新型组织蛋白;其余3个蛋白点在含量上相互间存在显著差异.本研究根据组织差异蛋白的功能初步推测家蚕对浓核病毒(镇江株)感性差异与血液蛋白差异无显著相关,与围食膜蛋白差异可能有关.     

食品常见真菌毒素的危害及其防止措施

真菌毒素是一类由真菌产生的普遍存在的化合物。危害人类健康的真菌毒素主要源于曲霉Aspergillus、青霉Penicil-lium、麦角菌属Claviceps和镰刀菌Fusarium等产生的次生代谢物。从全球范围来看,粮食安全问题经常是由谷物、坚果、水果和绿色咖啡豆上的真菌毒素造成的。其中,玉米和花生仁中的黄曲霉素经常超过安全阈值。在以这些食物作为主食的地处温暖和潮湿气候国家的消费者特别容易食用到黄曲霉素污染的食物。而真菌毒素往往会引起人类和动物急性中毒、慢性中毒和致癌。其不可避免地、广泛地、持续地影响着全球人类的健康。目前的防止措施包括选用抗真菌植物、采用适当方法贮藏食物以及食用绿色蔬菜等来预防癌症等。本文阐述了食品中常见真菌毒素的污染情况及其毒性,并对常用防止措施进行了综述,以期为食品真菌毒素防控工作提供参考。     

小型猪腹壁拉链模型的建立

目的建立小型猪腹壁拉链模型并对其生物学特性进行研究,为教学和科研工作的开展提供便利的研究工具。方法通过外科手术的方法将定制的生物拉链固定于小型猪体表,建立小型猪腹壁拉链模型,观察小型猪的体征和精神状态,利用全自动血液分析仪及尿液分析仪对其血液和尿液生化指标进行动态检测。结果模型建立后7-49d小型猪的活动、精神状态良好,其生理生化指标表现稳定,与对照相比无显著变化。结论本课题组建立的小型猪腹壁拉链模型建立后7-49d体征、精神状况良好,生理生化指标稳定,可以推广应用于相关的科研、教学试验。     

基于3种香茶菜属药用植物高效液相色谱的主成分分析

对香茶菜(Isodon amethystoides)、显脉香茶菜(I.nervosa)和大萼香茶菜(I.macrocalyx)不同产地和器官(根、茎、叶)共21个样品进行高效液相色谱分析,以出峰时间-峰面积为指标,以样品为对象进行主成分分析,比较不同样品间的差异程度。结果发现,(1)香茶菜、大萼香茶菜、显脉香茶菜在高效液相色谱上虽然有一定的差异,但这种差异并不明显;(2)基于HPLC显示的香茶菜不同种群间差异比三个种间的差异更为明显,说明不同产地对香茶菜属植物样品的植化组成有很大的影响,不同种在植化上的相似性使它们在一定程度上可以作为替代药材;(3)研究反映出基于高效液相色谱的PCA在反映不同样品植物化学组成差异程度上具有应用价值。     

Permeability of the infective juveniles of Steinernema carpocapsae to glycerol during osmotic dehydration and its effect on biochemical adaptation and energy metabolism

Permeability of the sheath and cuticle of the infective juveniles (IJs) of Steinernema carpocapsae to glycerol and its effect on biochemical adaptation of the IJs to osmotic dehydration were examined by incubating both sheathed and exsheathed IJs in glycerol-d5 solution then monitoring the changes in levels of deuterium labelled and non-labelled glycerol and trehalose. Energy metabolism of the IJs during osmotic dehydration and subsequent rehydration and the effect of the permeated glycerol on this process were investigated by examining and comparing the changes in mean dry weight and key biochemical composition of the IJs dehydrated in glycerol and sodium chloride solutions. The results show: (1) similarly to evaporative dehydration, osmotic dehydration induces IJs to synthesise the protectants glycerol and trehalose; (2) glycerol permeates the sheath and the cuticle into the body of IJs during dehydration in glycerol solution. Part of the permeated glycerol plays a role as protectant like that synthesised by IJs from their energy reserve materials while part is incorporated into trehalose; (3) the sheath reduces the rate of permeation of glycerol and therefore affects the equilibrium glycerol and trehalose levels of the IJs and also the time needed to reach the equilibrium levels; (4) the reduction in mean dry weight and lipids of the IJs during dehydration in glycerol solution is substantially less than those dehydrated in sodium chloride solution. Both the total protectant level and the ratio of glycerol to trehalose of the IJs dehydrated in glycerol solution are higher than those dehydrated in sodium chloride solution; (5) glycogen reserves of the IJs play a role as a buffer reservoir of the protectants during both dehydration and rehydration but the principal sources of the protectants during dehydration are more likely to be lipids and proteins rather than glycogen.