Sequential and compartmentalized action of Rabs,SNAREs, and MAL in the apical delivery of fusiform vesicles in urothelial umbrella cells

Uroplakins (UPs) are major differentiation products of urothelial umbrella cells and play important roles in forming the permeability barrier and in the expansion/stabilization of the apical membrane. Further, UPIa serves as a uropathogenic Escherichia coli receptor. Although it is understood that UPs are delivered to the apical membrane via fusiform vesicles (FVs), the mechanisms that regulate this exocytic pathway remain poorly understood. Immunomicroscopy of normal and mutant mouse urothelia show that the UP-delivering FVs contained Rab8/11 and Rab27b/Slac2-a, which mediate apical transport along actin filaments. Subsequently a Rab27b/Slp2-a complex mediated FV–membrane anchorage before SNARE-mediated and MAL-facilitated apical fusion. We also show that keratin 20 (K20), which forms a chicken-wire network ∼200 nm below the apical membrane and has hole sizes allowing FV passage, defines a subapical compartment containing FVs primed and strategically located for fusion. Finally, we show that Rab8/11 and Rab27b function in the same pathway, Rab27b knockout leads to uroplakin and Slp2-a destabilization, and Rab27b works upstream from MAL. These data support a unifying model in which UP cargoes are targeted for apical insertion via sequential interactions with Rabs and their effectors, SNAREs and MAL, and in which K20 plays a key role in regulating vesicular trafficking.     

Construction of an efficient genomic editing system with CRISPR/Cas9 in the vector mosquito Aedes albopictus

Aedes (Stegomyia) albopictus, also known as the Asian tiger mosquito, is a mosquito which originated in Asia. In recent years, it has become increasingly rampant throughout the world. This mosquito can transmit several arboviruses, including dengue, Zika and chikungunya viruses, and is considered a public health threat. Despite the urgent need of genome engineering to analyze specific gene functions, progress in genetical manipulation of Ae. albopictus has been slow due to a lack of efficient methods and genetic markers. In the present study, we established targeted disruptions in two genes, kynurenine hydroxylase (kh) and dopachrome conversion enzyme (yellow), to analyze the feasibility of generating visible phenotypes with genome editing by the clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated protein 9 (Cas9) system in Ae. albopictus. Following Cas9 single guide RNA ribonucleoprotein injection into the posterior end of pre-blastoderm embryos, 30%-50% of fertile survivors produced alleles that failed to complement existing kh and yellow mutations. Complete eye and body pigmentation defects were readily observed in GI pupae and adults, indicating successful generation of highly heritable mutations. We conclude that the CRISPR/Cas9-mediated gene editing system can be used mAe. albopictus and that it can be adopted as an efficient tool for genome-scale analysis and biological study.     

新媒体时代下医学生手机依赖与人际关系的相关性研究

目的:了解新媒体背景下的医学生手机依赖与人际关系现状及两者的相关性,为相关部门制定干预策略提供依据。方法:采取方便抽样的方法抽取样本,通过手机依赖和人际关系量表开展匿名问卷调查,SPSS19.0软件进行数据处理。结果:医学生中手机依赖阳性检出率为35.81%,人际交往敏感检出率为33.78%。手机依赖组的人际关系得分明显高于非手机依赖组(P0.05);同时,手机依赖量表与人际关系量表呈正相关(P0.05)。多元线性回归分析结果显示身体锻炼频率对手机依赖有反向预测作用,人际关系中的交际与交友维度对手机依赖有正向预测作用(P0.05)。结论:医学生手机依赖与人际交往情况形势严峻、不容乐观,社会各界应关注医学生手机依赖及其引发的系列公共卫生问题,多层面、多角度进行干预。     

水稻耐储藏特性三年动态鉴定与QTL分析

种子耐储藏特性是粮食作物的特殊农艺性状之一, 耐储藏性能对种子生产和种质资源保存有重要意义。以粳型超级稻龙稻5 (LD5)和高产籼稻中优早8 (ZYZ8)杂交衍生的重组自交系(RILs)群体(共180个株系)为实验材料, 自然高温高湿条件下放置1年、2年和3年后, 对不同储藏时段种子发芽率进行比较, 并利用223个分子标记的遗传图谱进行动态QTL鉴定。结果表明, 不同储藏时段龙稻5的发芽率均显著低于中优早8, 株系间耐储性存在较大差异; 不同储藏时段发芽率显著相关, 相邻存储时段发芽率关系紧密。共检测到17个耐储性相关的QTLs, 3个老化时段分别检测到5、4和3个, 检测到5个动态条件QTLs, 单一QTL解释5.60%-32.76%的表型变异, 加性效应在-16.78%-16.95%范围内。主效QTL簇qSSC2、qSSC6、qSSC7和qSSC8能调控不同储藏时段的发芽率, qSSC6具有明显降低发芽率的效应。共检测到26对上位性互作位点, 主效QTL qSS1和qSS4参与上位性互作, 这表明上位性互作是调控耐储藏性状的重要遗传组成。研究结果为水稻(Oryza sativa)耐储性相关QTL的精细定位奠定基础, 同时丰富了耐储性分子标记辅助选择育种的基因资源。     

iQPR法处理水对SD鼠生物安全性的研究

iQPR技术处理污水是一项新型尖端的技术,此技术可以成功降低污水乃至受到污染的地下水中的各种污染指标。但是,iQPR技术处理污水尤其是地下水是否存在潜在的生物安全性问题有待于进一步研究。因此,为评估iQPR技术对生物安全性的影响,本研究首先分析了三种不同iQPR法处理水的水质成分;其次系统研究了iQPR水对SD鼠在个体水平、组织水平和病理形态学损伤的研究。研究表明:iQPR处理的水质成分较对照组普通饮用水好,在个体组织水平检测未见异常,尽管其中一组iQPR处理水造成了SD鼠的脾小体增大,但是可能的原因是水处理环节存在微生物污染现象,因此,初步认定此技术未造成SD大鼠的个体损伤。本研究为揭示iQPR处理的水对生物体的安全性评价提供一个理论依据。     

广西红壤土壤微生物溶磷基因的克隆与表达

本研究对广西红壤土壤微生物溶磷基因进行克隆,并在大肠杆菌中诱导表达。从广西红壤中提取宏基因组DNA,克隆了GabY基因,构建大肠杆菌原核表达载体p ETDuet-1,转化大肠杆菌DH5α得到重组工程菌。通过酶切验证重组体。重组体在诱导条件下24 h对难溶矿物质磷Ca_3(PO_4)_2的溶磷量为(40.73±1.32)μg/mL,而对照组仅为(5.11±0.08)μg/mL。重组体和对照随着诱导时间的增加,培养基pH值均有下降趋势,但重组体培养基pH值变化显著大于对照组,产酸能力显著优于对照组。证明成功克隆得到土壤微生物溶磷基因,并在大肠杆菌中得到表达。     

微波法和超声波法提取绿穗苋多糖响应面优化研究

绿穗苋是一种药食兼用作物,其中多糖成份具有很高的药食价值。本研究以绿穗苋地上部分为材料,以微波和超声波两种方法对绿穗苋多糖进行提取,并通过响应面分析法对提取进行优化,确定最佳工艺,通过水提醇沉的方法得到绿穗苋粗多糖。综合比较两种提取工艺,提取效果最佳工艺为微波提取法。其条件为:提取时间41.42 min,提取功率211.65 W,料液比1:33.338 (g/mL),实际得率为13.25%。该研究结果促进绿穗苋资源的有效利用,为绿穗苋多糖的提取工艺及产品的开发利用提供理论依据。     

Genome-wide dissection of segregation distortion using multiple inter-subspecific crosses in rice

Mendelian inheritance can ensure equal segregation of alleles from parents to offspring, which provides fundamental basis for genetics and molecular biology. Segregation distortion(SD) leads to preferential transmission of certain alleles from generation to generation. Such violation of Mendelian genetic principle is often accompanied by reproductive isolation and eventually speciation. Although SD is observed in a wide range of species from plants to animals, genome-wide dissection of such biased transmission of gametes is rare. Using nine inter-subspecific rice crosses, a genome-wide screen for SD loci is performed, which reveals 61 single-locus quantitative trait loci and 194 digenic interactions showing distorted transmission ratio, among which 24 new SD loci are identified. Biased transmission of alleles is observed in all nine crosses, suggesting that SD exists extensively in rice populations. 72.13% distorted regions are repeatedly detected in multiple populations, and the most prevalent SD hotspot that observed in eight populations is mapped to chromosome 3. Xian alleles are transmitted at higher frequencies than geng alleles in inter-subspecific crosses, which change the genetic composition of the rice populations. Epistatic interaction contributes significantly to the deviation of Mendelian segregation at the whole-genome level in rice, which is distinct from that in animals. These results provide an extensive archive for investigating the genetic basis of SD in rice, which have significant implications in understanding the reproductive isolation and formation of inter-subspecific barriers during the evolution.     

心肌纤维化的信号转导机制及新型抑制剂的研究进展

心肌纤维化(myocardial fibrosis, MF)是心肌重构发生的重要病理过程,能够引起心脏衰竭甚至死亡。心肌组织中成纤维细胞异常增殖并转化为肌成纤维细胞以及心肌细胞外基质代谢紊乱导致沉积是心肌纤维化形成的主要病理基础。心肌纤维化发生的分子机制较复杂,已发现多种信号通路参与了心肌纤维化的发生。该文主要对参与调控心肌纤维化的信号转导机制进行了综述,并对新型信号抑制剂的研究进展进行了小结。