肽转运载体的分子特征及其分布

动物体内的肽转运载体目前发现的至少有五种,其中研究最为广泛的是：PepT1和PepT2。PepT1和PepT2都是依质子的寡肽转运载体(POT)家族的成员。PepT1是低亲和力／高容量的肽载体,PepT2高亲和力／低容量的肽载体。PepT1主要在消化道中表达,在肾脏中也有微弱的表达;PepT2主要在肾脏中表达。这些肽载体的分子结构特征主要有：(1)有12个假想的穿膜区,在9区和10区之间有一大的胞外环,且所有穿膜区内的序列都高度保守,胞外环上的序列保守的很少;(2)被编码的蛋白上有多个N-糖基化和蛋白激酶的识别位点,它们可能参与肽转运的调控;(3)PepT1上的His-57和PepT2上的His-87是最关键的组氨酸残基,它们可能是转运蛋白发挥吸收功能时最关键的结合位点;(4)不同动物肽转运蛋白的氨基酸范围在707到729之间,且不同动物相同器官肽转运载体的同源性高(大约80％),同种动物不同器官肽转运载体的同源性低(大约50％)。了解肽载体的分子特征和组织分布,可以更好地理解肽吸收的分子机制并有利于肽类药物的研发。     

四种冬季水生植物组合对富营养化水体的净化效果

选择10种耐低温的水生植物构建4种植物组合,研究了冬季低温环境下不同水生植物组合对富营养化水体的净化效果.结果表明:组合1[常绿水生鸢尾(Iris hexagonus Hybrid)*羊蹄(Rumex japonicus)+金叶“金钱蒲”(Acorus gramineus“Ogan”)+反曲灯心草“蓝箭”(Juncus inflexus“Blue Arrows”)]4种植物均能在试验富营养化水体中茂盛生长,且对TN、NOx-N、NH4-N和TP的去除率分别为47.8％、52.2％、32.4％和70.1％;组合2[常绿水生鸢尾+羊蹄+金叶“金钱蒲”+大苞萱草(Hemerocallis middendorfii)]4种植物也都能在试验富营养化水体中存活,并且有一定量的生长,对TN、NOx-N、NH4-N和TP的去除率分别为44.2％、58.5％、34.6％和67.8％;而未种植物的对照对TN、NOx-N、NH4-N和TP的去除率分别为40.0％、25.9％、27.3％和64.5％;组合1和2对富营养化水体有较好的净化效果.组合3和组合4中由于吊兰(Chlorophytum comosum)和三穗薹草(Carex tristachya)等植物长势较差,仅对NOx-N具有较明显去除能力,对其他指标去除效果不明显.通过这些水生植物在富营养化水体中生长特性和对营养元素的去除能力,发现冬季组合1和2的净化效果较好,是低温条件下适宜的浮床植物组合形式.     

山梨猕猴桃与中华猕猴桃种间杂交后代果实性状的遗传倾向分析

以山梨猕猴桃(Actinidia rufa(Siebold and Zuccarini)Planchon ex Miquel)品种‘63101’和中华猕猴桃(Actinidia chinensis var.chinensis C. F. Liang)品种‘磨山雄7号’杂交产生的400余株F₁代个体为材料,连续2～3年对其果实品质和外观特点的9个性状进行测定,分析它们的遗传规律。结果显示,子代群体单果质量连续3年低于双亲中值,遗传传递力连续3年低于100%,优势率连续3年均为负值;可溶性固形物子代中值除2015年略低外,其余2年均高于亲中值12.3%;优势率均为正值,遗传传递力均大于100%;子代果实形状出现分离,与母本圆柱形相同的比例占82%;果皮毛被中,偏离母本的有毛比例为60%;果皮颜色出现绿色、绿褐色、褐色的分离,比例为1∶1.5∶1;果实肩部形状多与母本的方形相似,占比为63%;果肉颜色也多与母本的绿色相似,占比为86%;果实风味中,与父本和母本相似的表型占比均为32%,口感酸的占比18%,超低亲极酸占比1%,超高亲浓甜占比9%。说明杂交子代单果质量和可溶...     

钾肥及与秸秆配施对紫色土作物产量和微生物群落结构的影响

基于紫色菜园土壤莴笋-白菜-甜玉米轮作4年12季连续定位施钾肥试验,研究不同种类钾肥及用量对重庆地区紫色菜园土上作物产量和土壤微生物群落结构的影响,为该地区作物生产确定合理的施钾技术提供科学依据。结果表明,在施用氮磷肥基础上,不同钾肥处理莴笋、白菜、甜玉米产量均显著提高;从3种作物4年平均产量和钾肥效益看,以单施适量化学钾(K_2)、单施适量秸秆钾(M_2)和秸秆还田配施少量化学钾(K_1+M_1)的施钾肥模式对白菜的增产效果好,增产28.05%—30.27%;M_2和K_1+M_1施钾肥模式对莴笋的增产效果较好,增产13.89%和13.81%;K_1+M_1施钾肥模式对甜玉米的增产效果最好,增产15.10%,表明秸秆还田配施少量化学钾(K_1+M_1)模式在供试3种作物生产中具有很好的应用前景。不同施钾处理对土壤微生物群落结构及丰度均会产生一定影响,以秸秆还田配施少量化学钾肥(K_1+M_1)对提高土壤细菌、真菌、革兰氏阳性菌的作用较好,并显著增加土壤微生物总PLFA含量,缓解对土壤微生物生存环境的胁迫,说明K_1+M_1能为土壤微生物的生长繁育创造有利环境。莴笋、白菜、甜玉米3种作物4年平均产量与土壤细菌、真菌、革兰氏阳性菌和革兰氏阴性菌数量间存在显著相关性(P0.05),而不同钾肥处理对土壤微生物特征具有一定影响,说明施钾肥可能会调节土壤微生物特征,从而促进作物增产。     

Genome‐wide analysis of Nilaparvata lugens nymphal responses to high‐density and low‐quality rice hosts

The brown planthopper (BPH) Nilaparvata lugens is an economically important pest on rice plants. In this study, the higher population density and yellow‐ripe stage of rice plants were used to construct adverse survival conditions (ASC) against BPH nymphs. Simultaneously, the low population density and tillering stage of rice plants were used to establish a suitable survival condition (SSC) as a control. Solexa/Illumina sequencing was used to identify genes of BPH nymphs responding to ASC. Significantly longer duration development of BPH nymphs and significantly lower brachypterous ratio of BPH adults were observed by ASC compared with SSC. A total of 2 544 differentially expressed genes (DEGs) were obtained and analyzed by BLASTx, Gene Ontology and KEGG Orthology. Gene ontology analysis revealed that the DEGs were mainly involved in categories of cell, cell part, cellular process, binding, catalytic, organelle and metabolic processes. 1 138 DEGs having enzyme commission numbers were assigned to different metabolic pathways. The largest clusters were neurodegenerative diseases (137, 12.0%), followed by carbohydrate metabolism (113, 9.9%), amino acid metabolism (94, 8.3%), nucleotide metabolism (76, 6.7%), energy metabolism (64, 5.6%), translation (60, 5.3%), lipid metabolism (58, 5.1%), and folding, sorting and degradation (52, 4.6%). Expressing profile of 11 DEGs during eight nymphal developmental stages of BPH were analyzed by quantitative real‐time polymerase chain reaction. The 11 genes exhibited differential expression between ASC and SSC during at least one developmental stage. The DEGs identified in this study provide molecular proof of how BPH reconfigures its gene expression profile to adapt to overcrowding and low‐quality hosts.     

Isolation and characterization of <Emphasis Type="Italic">Paenibacillus polymyxa</Emphasis> LY214, a camptothecin-producing endophytic bacterium from <Emphasis Type="Italic">Camptotheca acuminata</Emphasis>

Camptothecin (CPT) is mainly produced and extracted from Camptotheca acuminata and Nothapodytes foetida for pharmaceutical use, i.e., the starting material for chemical conversion to the clinical CPT-type drugs. As the third largest plant anticancer drug, the heavy demand on CPT from global market leads to many research efforts to identify new sources for CPT production. Herein we report the isolation and characterization of a CPT-producing endophytic bacterium Paenibacillus polymyxa LY214 from Camptotheca acuminata. A 10.7 μg l^?1 of CPT was presented in the fermentation broth of P. polymyxa LY214. Its CPT production decreased sharply when the strain of the 2nd generation of P. polymyxa LY214 was cultured and fermented. However, the CPT production remained relatively constant from 2.8 μg l^?1 of the 2nd generation to 0.8 μg l^?1 of the 8th generation of P. polymyxa LY214 under optimized fermentation conditions. A 15- to 30-fold increase of CPT yield was observed when the optimized fermentation conditions, together with the addition of putative biosynthetic precursors of CPT and adsorbent resin XAD16, were applied to ferment the strains of the 7th and 8th generation of P. polymyxa LY214. Bioinformatics analysis of the relative species of P. polymyxa LY214 indicates its potential to produce CPT, which will be helpful to decipher the mysteries of CPT biosynthesis.     

Clopidogrel reduces lipopolysaccharide‐induced inflammation and neutrophil‐platelet aggregates in an experimental endotoxemic model

Platelet activation contributes to organs failure in inflammation and plays an important role in endotoxemia. Clopidogrel inhibits platelet aggregation and activation. However, the role of clopidogrel in modulating inflammatory progression of endotoxemia remains largely unexplored. Therefore, we investigated the role of clopidogrel on the activation of platelet and leukocytes in lipopolysaccharide (LPS)‐induced inflammation in mice. Animals were treated with clopidogrel or vehicle before LPS induction. The expression of neutrophil‐platelet aggregates and platelet activation and tissue factor was determined. Immunofluorescence was used to analyze platelet‐leukocyte interactions and tissue factor (TF) expression on leukocytes. Clopidogrel pretreatment markedly decreased lung damage, inhibited platelet‐neutrophil aggregates and TF expression. In addition, clopidogrel reduced thrombocytopenia and affected the number of circulating white blood cell in endotoxemia mice. Moreover, clopidogrel also reduced platelet shedding of CD40L and CD62P in endotoxemic mice. Taken together, clopidogrel played an important role through reducing platelet activation and inflammatory process in endotoxemia.     

转人工miRNA抗玉米粗缩病毒载体玉米的培育及其抗病能力

玉米是重要的粮食作物,水稻黑条矮缩病毒(RBSDV)是玉米粗缩病的病原,由其引起的玉米粗缩病给玉米生产造成重大损失。利用人工mi RNA构建抗病毒植物的技术已经在多种植物中被证明有效,但是在玉米中的尝试未见报道。实验根据玉米zea-mi R159a的前体序列和RBSDV基因组中编码功能蛋白的基因和基因沉默抑制子的序列信息设计引物,构建了用于沉默RBSDV编码基因和基因沉默抑制子的ami RNA(Artificial mi RNA)基因。构建p CAMBIA3301-121-ami RNA植物表达载体,利用农杆菌介导法转化玉米自交系综31(Z31)。对转基因玉米进行分子检测,选择mi RNA表达量高的纯合体株系进行自然发病实验,按0-4的分级标准调查玉米粗缩病的严重度。结果表明,转抗粗缩病毒人工mi RNA载体玉米纯合体株系的抗病表现好于野生型玉米,其中针对基因组6的S6-mi R159转基因玉米抗病情况较好。研究表明利用人工mi RNA技术构建抗病毒病玉米新品种是可行的。     

Antiinflammatory Activity of a Novel Folic Acid Targeted Conjugate of the mTOR Inhibitor Everolimus

Folate receptor (FR)-β has been identified as a promising target for antimacrophage and antiinflammatory therapies. In the present study, we investigated EC0565, a folic acid–derivative of everolimus, as a FR-specific inhibitor of the mammalian target of rapamycin (mTOR). Because of its amphiphilic nature, EC0565 was first evaluated for water solubility, critical micelle formation, stability in culture and FR-binding specificity. Using FR-expressing macrophages, the effect of EC0565 on mTOR signaling and cellular proliferation was studied. The pharmacokinetics, metabolism and bioavailability of EC0565 were studied in normal rats. The in vivo activity of EC0565 was assessed in rats with adjuvant arthritis, a “macrophage-rich” model with close resemblance to rheumatoid arthritis. EC0565 forms micellar aggregates in physiological buffers and demonstrates good water solubility as well as strong multivalent FR-binding capacity. EC0565 inhibited mTOR signaling in rat macrophages at nanomolar concentrations and induced G0/G1 cell cycle arrest in serum-starved RAW264.7 cells. Subcutaneously administered EC0565 in rats displayed good bioavailability and a relatively long half-life (~12 h). When given at 250 nmol/kg, EC0565 selectively inhibited proliferating cell nuclear antigen expression in thioglycollate-stimulated rat peritoneal cells. With limited dosing regimens, the antiarthritic activity of EC0565 was found superior to that of etanercept, everolimus and a nontargeted everolimus analog. The in vivo activity of EC0565 was also comparable to that of a folate-targeted aminopterin. Folate-targeted mTOR inhibition may be an effective way of suppressing activated macrophages in sites of inflammation, especially in nutrient-deprived conditions, such as in the arthritic joints. Further investigation and improvement upon the physical and biochemical properties of EC0565 are warranted.