从大容量噬菌体抗体库中筛选抗Acr蛋白人源单链抗体

目的:从天然的大容量噬菌体抗体库中筛选特异的抗结核分枝杆菌晶体蛋白( alpha-crystallin Acr)的人源抗体.方法:以结核分枝杆菌Acr蛋白包被免疫管,通过对噬菌体抗体库进行4轮“吸附-洗脱-扩增”的过程从大容量抗体库中筛选特异性抗结核分枝杆菌Acr蛋白的抗体,并对可变区序列进行了测序分析.将特异性的噬菌体抗体感染HB2151菌,经IPTG诱导表达,制备了抗结核分枝杆菌Acr蛋白的可溶性单链抗体;对其序列和抗原结合活性进行分析鉴定.结果:经过4轮筛选,获得了43个与结核分枝杆菌Acr蛋白结合的阳性克隆,其中29个特异结合的克隆;测序分析有26不同的可变区片段;通过可溶性单链抗体(scFv)表达筛选到14株特异性结合Acr蛋白的可溶性单链抗体克隆;经过基因测序,分析了可变区基因的亚群.成功制备了可溶性单链抗体.Westren blotting分析证实筛选的人源单链抗体能与天然蛋白结合.结论:利用单链大容量抗体库获得抗结核分枝杆菌Acr蛋白的噬菌体抗体并且成功制备抗结核分枝杆菌Acr天然蛋白的可溶性单链抗体,为今后的研究和应用奠定基础.     

Human Fidgetin is a microtubule severing the enzyme and minus-end depolymerase that regulates mitosis

Fidgetin is a member of the AAA protein superfamily with important roles in mammalian development. Here we show that human Fidgetin is a potent microtubule severing and depolymerizing the enzyme used to regulate mitotic spindle architecture, dynamics and anaphase A. In vitro, recombinant human Fidgetin severs taxol-stabilized microtubules along their length and promotes depolymerization, primarily from their minus-ends. In cells, human Fidgetin targets to centrosomes, and its depletion with siRNA significantly reduces the velocity of poleward tubulin flux and anaphase A chromatid-to-pole motion. In addition, the loss of Fidgetin induces a microtubule-dependent enlargement of mitotic centrosomes and an increase in the number and length of astral microtubules. Based on these data, we propose that human Fidgetin actively suppresses microtubule growth from and attachment to centrosomes.     

Socioeconomic Factors and All Cause and Cause-Specific Mortality among Older People in Latin America,India, and China: A Population-Based Cohort Study

Background

Even in low and middle income countries most deaths occur in older adults. In Europe, the effects of better education and home ownership upon mortality seem to persist into old age, but these effects may not generalise to LMICs. Reliable data on causes and determinants of mortality are lacking.

Methods and Findings

The vital status of 12,373 people aged 65 y and over was determined 3–5 y after baseline survey in sites in Latin America, India, and China. We report crude and standardised mortality rates, standardized mortality ratios comparing mortality experience with that in the United States, and estimated associations with socioeconomic factors using Cox''s proportional hazards regression. Cause-specific mortality fractions were estimated using the InterVA algorithm. Crude mortality rates varied from 27.3 to 70.0 per 1,000 person-years, a 3-fold variation persisting after standardisation for demographic and economic factors. Compared with the US, mortality was much higher in urban India and rural China, much lower in Peru, Venezuela, and urban Mexico, and similar in other sites. Mortality rates were higher among men, and increased with age. Adjusting for these effects, it was found that education, occupational attainment, assets, and pension receipt were all inversely associated with mortality, and food insecurity positively associated. Mutually adjusted, only education remained protective (pooled hazard ratio 0.93, 95% CI 0.89–0.98). Most deaths occurred at home, but, except in India, most individuals received medical attention during their final illness. Chronic diseases were the main causes of death, together with tuberculosis and liver disease, with stroke the leading cause in nearly all sites.

Conclusions

Education seems to have an important latent effect on mortality into late life. However, compositional differences in socioeconomic position do not explain differences in mortality between sites. Social protection for older people, and the effectiveness of health systems in preventing and treating chronic disease, may be as important as economic and human development. Please see later in the article for the Editors'' Summary     

黄土高原樟子松和落叶松与其他树种枯落叶混合分解对土壤的影响

通过采集树木枯落叶与土壤进行室内混合分解培养试验,研究了黄土高原常见的樟子松和落叶松与其他树种枯落叶混合分解对土壤性质的影响及存在的相互作用,从而为不同树木种间关系的探索和该地区人工纯林的混交改造提供科学指导。结果表明:12种枯落叶单一分解均明显提高了土壤脲酶(54%—110%)、脱氢酶(85%—288%)和磷酸酶(81%—301%)活性以及有机质(29%—55%)和碱解N(12%—49%)含量,但对土壤速效P含量和CEC的影响存在较大差异。综合而言,樟子松分别与白桦、刺槐、白榆、柠条和落叶松枯落叶混合分解在对土壤性质的影响中存在相互促进作用,而分别与小叶杨、沙棘、紫穗槐、侧柏和辽东栎枯落叶混合分解在对土壤性质的影响中存在相互抑制作用;落叶松分别与刺槐、白桦、小叶杨和紫穗槐枯落叶混合分解在对土壤性质的影响中存在相互促进作用,而分别与柠条、侧柏、辽东栎、沙棘、油松和白榆枯落叶混合分解在对土壤性质的影响中存在相互抑制作用。     

药物引起肾功能损伤的研究进展

肾脏是人体血流量丰富,耗氧量最大的主要脏器之一。药物主要是帮助机体对抗疾病,同时药物也可以引起组织器官的损伤。由于大部分药物及其代谢产物经由肾脏排出体外,大大增加了药物对肾功能损伤的机率。因此,研究药物对肾脏的毒性作用并弄清药物所导致的肾损伤的作用机制,就显得尤为重要。这对于开发新药和临床合理用药均有着重要的意义。本文就目前引起肾损伤药物和引起损伤的相关机制进行综述。     

Airway epithelial versus immune cell Stat1 function for innate defense against respiratory viral infection

The epithelial surface is often proposed to actively participate in host defense, but evidence that this is the case remains circumstantial. Similarly, respiratory paramyxoviral infections are a leading cause of serious respiratory disease, but the basis for host defense against severe illness is uncertain. Here we use a common mouse paramyxovirus (Sendai virus) to show that a prominent early event in respiratory paramyxoviral infection is activation of the IFN-signaling protein Stat1 in airway epithelial cells. Furthermore, Stat1-/- mice developed illness that resembled severe paramyxoviral respiratory infection in humans and was characterized by increased viral replication and neutrophilic inflammation in concert with overproduction of TNF-alpha and neutrophil chemokine CXCL2. Poor control of viral replication as well as TNF-alpha and CXCL2 overproduction were both mimicked by infection of Stat1-/- airway epithelial cells in culture. TNF-alpha drives the CXCL2 response, because it can be reversed by TNF-alpha blockade in vitro and in vivo. These findings pointed to an epithelial defect in Stat1-/- mice. Indeed, we next demonstrated that Stat1-/- mice that were reconstituted with wild-type bone marrow were still susceptible to infection with Sendai virus, whereas wild-type mice that received Stat1-/- bone marrow retained resistance to infection. The susceptible epithelial Stat1-/- chimeric mice also exhibited increased viral replication as well as excessive neutrophils, CXCL2, and TNF-alpha in the airspace. These findings provide some of the most definitive evidence to date for the critical role of barrier epithelial cells in innate immunity to common pathogens, particularly in controlling viral replication.     

海南黄牛HSF1 cDNA全长的克隆与序列分析

目的：根据人、小鼠HSF1cDNA保守区序列设计引物,通过PCR方法扩增海南黄牛HSF1cDNA,并进行序列分析。方法：利用RT-PCR、半巢式PCR以及3＇-RACE技术分段扩增得到了海南黄牛HSF1cDNA序列,测序正确后进行拼接。用DNAMAN 生物信息学软件分析海南黄牛HSF1 cDNA与赫里福德牛、人、小鼠同源性和海南黄牛HSF1蛋白的氨基酸组成、等电点、亲/疏水区等蛋白质性质,并根据各种动物HSF1蛋白绘制进化树。结果：（1）海南黄牛的HSF1 cDNA序列全长为1 993bp,包括150bp的5＇非翻译区,1 578bp的开放阅读框以及264bp（不含poly（A）尾）的3＇非翻译区,编码524个氨基酸,分子量为56.42 kD,等电点（pI）为 4.79。（2）海南黄牛的HSF1 cDNA与赫里福德牛、小鼠和人HSF1 cDNA的同源性分别为98.99 %、81.78 %、87.82 %,相应编码蛋白氨基酸序列的同源性分别为98.86 %、83.84 %、89.06 %,其中N-末端和C-末端高度保守,而中间区域存在缺失或替换。（3）根据氨基酸序列构建不同动物HSF1蛋白的进化树,与采用经典遗传分类法构建的进化树基本一致。结论：首次克隆了海南黄牛 HSF1 cDNA全长,分析表明：海南黄牛HSF1蛋白是亲水性蛋白,在8种动物中,其同源性大于73 %,高度保守。海南黄牛与赫里福德牛HSF1蛋白同源性高达98.86 %,在三聚体化区域、转录调节域和激活域存在6个位点的单氨基酸突变,这些发现为进一步揭示海南黄牛抗热性状形成的分子机制提供了重要依据。     

对一株铜绿假单胞菌22种耐药基因的研究

目的研究铜绿假单胞菌多重耐药情况和相关机制。方法采用聚合酶链反应（PCR）法对一株多重耐药铜绿假单胞菌进行β-内酰胺酶基因、氨基糖苷类修饰酶（AMEs）基因、喹诺酮类耐药基因、耐消毒剂基因（qacE△1-sul1）和整合酶基因检测,并对VIM基因进行了测序。结果PCR扩增结果显示该菌株aac（6′）-Ⅰb、blaCARB、gyrA、oprD2、ant（2″）-Ⅰ、qacE△1-sul1、blaIMP-I、blaTEM、blaVEB、aac（3）-Ⅱ、ant（3″）-Ⅰ、intⅠ1、blaVIM基因均为阳性,而aac（3）-Ⅰ、aac（6′）-Ⅱ、blaGES、blaGIM、blaOXA-10群、blaPER、blaSPM、blaSHV、blaDHA基因均为阴性,VIM基因扩增产物测序后经BLAST同源性分析表明为VIM-2型。结论铜绿假单胞菌存在多重耐药基因。管壁涂有季胺类、双胍类消毒剂和磺胺的II代导管的抑菌效果需重新评价。     

The identity of <Emphasis Type="Italic">Zamia katzeriana</Emphasis> and <Emphasis Type="Italic">Z. verschaffeltii</Emphasis> (Zamiaceae)

The morphological variation of recently described species (also the older taxa) of Zamia distributed within southeastern Mexico that have wide, coriaceous leaflets is analyzed. The complex is designated as the Z. katzeriana species complex in reference to an historic collection of this name, which is also the earliest named species in the complex. The complex consists of Z. cremnophila, Z. lacandona, Z. splendens and Z. purpurea. During the 1990’s, Z. splendens was separately synonymized under Z. katzeriana and Z. verschaffeltii, both collected in Mexico by German collectors during the 18th century, but the precise locality is unknown. Information concerning the incomplete holotype of Z. verschaffeltii is particularly ambiguous, and the possible type locality suggested by Schuster, Socorro, is imprecise, thus generating taxonomic confusion. Morphometric characterization and discriminant analysis of the contemporary and historic collections (i.e., Z. katzeriana and Z. verschaffeltii) included all the known populations (11) and individuals (115) of the complex throughout its range. The results show that Z. verschaffeltii is not morphometrically related to any of the species in the complex and that Z. splendens should be considered a synonym of Z. katzeriana.

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Resumen  En este artículo se caracteriza la variación morfológica de las especies de un complejo de Zamia, que presentan folíolos anchos y coriáceos con distribución en el sureste de México, la mayoría de reciente descripción. El complejo es denominado Z. katzeriana, por una colección histórica de este nombre, y consiste de Z. cremnophila, Z. lacandona, Z. splendens y Z. purpurea. Zamia splendens fue sinonimizada separadamente bajo dos colecciones históricas Z. katzeriana y Z. verschaffeltii, de las cuales se desconoce con exactitud su lugar de procedencia, excepto que fueron colectadas en México por botánicos alemanes del siglo XVIII. La información, en especial sobre el holotipo incompleto de Z. verschaffeltii es muy ambigua y la posible localidad tipo mencionada por Schuster, Socorro, es imprecisa, lo cual ha generado confusiones taxonómicas. La caracterización de la variación morfométrica incluyó a todas las poblaciones (11) e individuos (115) conocidas actualmente para el complejo en todo su rango de distribución en México. El análisis discriminante incluyendo a las colecciones históricas, determina que Z. verschaffeltii no está relacionado morfométricamente con ninguna especie del complejo y Z. katzeriana está estrechamente correlacionada con Z. splendens.

     

硝酸盐供应对玉米侧根生长的影响

以两个玉米(Zea mays L.)自交系478和Wu312为研究材料,采用琼脂培养方法,研究不同浓度NO-3对侧根生长的影响.结果表明,在外部浓度0.01～1.0mmol/L范围内,NO-3供应能显著增加侧根的长度及根生物量.但当NO-3供应超过1.0 mmo1/L后,侧根长度开始下降.当NO-3供应分别在超过5.0(Wu312)与10(478)mmol/L后,侧根密度显著下降.在10 mmol/LNO-3下,Wu312的侧根生长几乎完全被抑制.而478在20 mmol/L时,侧根密度仍可达到其最大值的30%(主根)～50%(胚根).植株地上部全氮及硝酸盐含量随NO-3供应的增加而升高,二者与侧根长度、侧根密度及冠根比的数学函数关系相同.