基于微流控芯片电泳的番茄黄化曲叶病毒快速检测

【目的】探索微流控芯片电泳方法在PCR产物检测方面的效果,并建立针对番茄黄化曲叶病毒（Tomato yellow leaf curl virus,TYLCV）的微流控芯片电泳检测方法,弥补琼脂糖凝胶电泳方法在试剂消耗、所用时间、安全性方面的缺陷。【方法】通过对TYLCV的基因组进行分析后,在该基因组中相对稳定的位置设计引物,同时兼顾引用已被研究者研究过的引物,对这些选择的引物进行特异性、稳定性、灵敏度等方面的验证,筛选出用于后续试验的引物。选择DNA标准物φX174/BsuR I(Hae Ⅲ) marker,分别进行琼脂糖凝胶电泳和微流控芯片电泳,对二者在耗材、耗时和灵敏度方面进行比较,确定微流控芯片电泳在核酸检测方面的应用价值。利用筛选出的其中1对引物对番茄叶片的实际样品进行PCR扩增,随后通过微流控芯片电泳对其进行检测,以此探讨微流控芯片电泳在病毒检测方面的检测效果。【结果】共筛选出14对TYLCV备用引物,其中2对引自文献,12对为本文设计,每对引物均可满足微流控芯片检测要求。选择其中1对引物TYLCV-T作为随后的研究对象。利用琼脂糖凝胶电泳和微流控芯片电泳对DNA标准物检测,结果表明微流控芯片电泳在耗时方面不足琼脂糖凝胶电泳的1/10,约为13 min,试剂消耗为琼脂糖凝胶电泳的1/8,检测灵敏度方面至少比琼脂糖凝胶电泳高103倍,根据DNA标准物原液浓度计算可知,微流控芯片电泳至少可准确检测到浓度为5×10-6 μg?μL-1的核酸样品。利用微流控芯片电泳对TYLCV-T扩增的TYLCV PCR产物进行检测,将检测峰值图与DNA标准物的峰值图时间比较,就可判断出产物峰的大小范围。【结论】筛选出的关于TYLCV的备用引物可作为进一步研究微流控芯片技术在该病毒检测方面的基础;通过将琼脂糖凝胶电泳和微流控芯片电泳进行比较,确立了后者在核酸检测方面的应用价值;通过微流控芯片电泳对TYLCV PCR产物的检测,建立了基于微流控芯片电泳的TYLCV快速检测方法,为TYLCV的快速检测提供新的技术支持。     

更昔洛韦眼用凝胶和阿昔洛韦滴眼液治疗单疱病毒性角膜炎疗效观察

目的探讨更昔洛韦眼用凝胶和阿昔洛韦滴眼液治疗单疱病毒性角膜炎的临床效果.方法单疱病毒性角膜炎患者104例均为单眼发病.根据治疗方案分为两组,采用阿昔洛韦滴眼液治疗患者52例为对照组,采用更昔洛韦眼用凝胶治疗患者52例为观察组,疗程3周,随访1a,比较两组患者的治疗时间、治疗效果、不良反应及复发情况.结果观察组患者浅层型、深层型、混合型单疱病毒性角膜炎治疗时间均明显少于对照组,观察组患者总有效率明显高于对照组,观察组患者不良反应发生率、复发率均明显低于对照组,差异具有统计学意义（P〈0.05）.结论更昔洛韦眼用凝胶和阿昔洛韦滴眼液均是治疗单疱病毒性角膜炎的有效药物.更昔洛韦眼用凝胶的治疗效果更好,可明显缩短患者的治疗时间,不良反应少,且复发率低,值得临床推广使用.     

稻米胶稠度的遗传研究

 采用6×6完全双列杂交和改良单籽粒分析法，对胶稠度的遗传和基因作用进行了研究。试验结果表明，硬对中等或软胶稠度、中等对软胶稠度表现显性。亲子方差和协方差(Vr-Wr)分析指出胶稠度受到包括一对主效基因和若干微效基因(修饰基因)的控制，基因的平均显性作用为不完全显性。遗传参数分析表明在胶稠度的遗传中基因的显性作用和加性作用是重要的，狭义遗传力较高(h²(ns) =0.78)．本文还讨论了在胶稠度改良中亲本选配和分离世代早期选择和重要性。     

高氟低碘对老年大鼠肝脏细胞DNA损伤的影响

[目的]研究高氟低碘对大鼠肝脏细胞DNA损伤的影响。[方法]离乳Wistar大鼠21只,随机分为4组：①CK（5只）,饲喂大鼠标准饲料,饮自来水;②高氟组（5只）,饲喂大鼠正常饲料,饮100mg/L氟化钠（NaF）去离子水;③低碘组（5只）,饲喂低碘饲料（定做）,饮去离子水;④高氟低碘组（6只）,饲喂低碘饲料,饮100mg/L氟化钠（NaF）去离子水。在大鼠20月龄时,处死取肝脏组织,检查其DNA的损伤情况。[结果]与①CK相比,②、③和④的老年大鼠肝细胞拖尾率显著增高（P〈0.01）;②、③和④的老年大鼠肝细胞的拖尾长度亦显著增长（P〈0.01）;肝细胞DNA损伤分级结果显示,②（高氟组）老年大鼠肝细胞DNA损伤主要是Ⅱ级、Ⅲ级。③（低碘组）肝细胞DNA几乎全部损伤,主要是Ⅲ级损伤。④（高氟低碘组）的细胞损伤以Ⅲ级损伤为主。[结论]高氟、低碘、高氟低碘都影响了大鼠肝脏细胞DNA的损伤。     

Transgenic sorghum with suppressed synthesis of kafirin subclasses: Effects on flour and dough rheological characteristics

Arising from work showing that conventionally bred high protein digestibility sorghum types have improved flour and dough functionality, the flour and dough properties of transgenic biofortified sorghum lines with increased protein digestibility and high lysine content (TG-HD) resulting from suppressed synthesis of several kafirin subclasses, especially the cysteine-rich γ-kafirin, were studied. TG-HD sorghums had higher flour water solubility at 30 °C (p < 0.05) and much higher paste viscosity (41% higher) than their null controls (NC). TG-HD doughs were twice as strong as their NC and dynamic rheological analysis indicated that the TG doughs were somewhat more elastic up to 90 °C. CLSM of doughs and pastes indicated that TG-HD had a less compact endosperm protein matrix surround the starch compared to their NC. The improved flour and dough functional properties of the TG-HD sorghums seem to be caused by reduced endosperm compactness resulting from suppression of synthesis of several kafirin subclasses which modifies protein body and protein matrix structure, and to improved protein-starch interaction through hydrogen bonding specifically caused by reduction in the level of the hydrophobic γ-kafirin. The improved flour functionality of these transgenic biofortified sorghums can increase their commercial utility by complementing their improved nutritional quality.     

人参止血成分的化学研究

本文采用逆流分布和凝胶色谱(Sephadex LH-20和CM-sephedex C-25)从人参根中分离出一种具有止血活性的化合物,经鉴定,该化合物为β-N-草酰基-L-α,β-二氨基丙酸。     

不同NSI值脱腥大豆粉凝胶形成性的评价

本文通过水溶大豆粉凝固性试验对几种不同ＮＳＩ值大豆粉凝胶形成性，以考察不同工艺生产的大豆食品中物理添加特性，证明了不同剂量高频处理工艺生产不同ＮＳＩ值大豆粉凝胶形成性情况。同时证明只有ＮＳＩ值＞５０％时，具有良好的凝胶性。     

Susceptibility to Coolness at the Young Microspore Stage under High Nitrogen Supply in Rice (Oryza Sativa L.). Proteome Analysis of Mature Anthers

Abstract

In vitro pollen germination experiment using agar plates showed that the growth under high nitrogen conditions enhanced the damage to pollen germination ability caused by the cooling at the young microspore stage. To clarify the physiological factors related to this damage to pollen germination, we performed the comparative proteome analysis of mature anthers and identified proteins that were changed by high nitrogen conditions or high nitrogen plus cooling conditions. Proteins were extracted from mature anther samples and separated by two-dimensional polyacrylamide gel electrophoresis. By comparing anther protein maps of the samples collected from the plants grown under standard nitrogen conditions, high nitrogen conditions and high nitrogen plus cooling conditions, we found 11 protein spots, which varied with the treatment. These protein spots were identified based on the rice proteome database and/or peptide mass fingerprinting (PMF) analysis after digestion with trypsin. Digested samples were analyzed by matrix-assisted laser desorption/ionization-time flight mass spectrometry to produce PMF data. Database searches using these PMF data revealed the identities of 9 proteins. Seven of these proteins were polypeptides involved in cell elongation, stress responses and sugar metabolism. The relation between the fluctuations of these proteins and the decrease in pollen germination are discussed.     

水稻花粉总蛋白质双向凝胶电泳方法建立及应用

采用石英砂加液氮研磨方法破碎花粉,然后用两种不同的方法提取水稻花粉总蛋白质,之后采用固相pH梯度等电聚焦/SDS PAGE 双向凝胶电泳对红莲型细胞质雄性不育水稻单核期花粉总蛋白质进行了分离。通过银染显色, PDQuest 2DE软件可识别约1 000~1 500个蛋白质点。其中TCA 丙酮提取法更适合提取花粉总蛋白质进行双向凝胶电泳分离,并可获得分辨率和重复性较好的双向电泳图谱。对红莲型细胞质雄性不育水稻的不育系单核期和二核期花粉总蛋白进行了双向电泳分离,通过比较两个时期花粉蛋白质电泳图谱后发现,二核期的花粉蛋白质较单核期花粉蛋白质数目减少,并且部分蛋白质表达量下降。