FRIL蛋白体外维持脐血造血干/祖细胞的作用及细胞周期调控基因的表达

为探讨新的豆类凝集素(Flt3 receptor-interacting lectin,FRIL)体外维持脐血CD34^ 细胞的作用以及维持过程中细胞周期调控基因HTm4及HTm4S mRNA的表达及意义,我们利用FRIL维持培养脐血CD34^ 细胞,对其增殖曲线、细胞周期及集落形成能力进行常规分析,并用半定量RT—PCR法分别测定FRIL体外维持不同时间后脐血CD34^ 细胞中周期调控基因HTm4及HTm4S mRNA的表达变化。结果显示,FRIL培养的CD34^ 造血干／祖细胞的增殖趋势平缓,整个培养期间细胞增殖倍数不超过起始的3倍：14d之前,FRIL培养细胞的高增殖潜能集落形成细胞(HPP—CFC)形成集落数与FL组无差别,其后则维持高于FL的情况。细胞周期分析则显示,在28d的培养过程内,利用FRIL培养的细胞始终有80％以上维持在G0期;而周期调控基因HTm4及HTm4S在刚分离的脐血CD34^ 细胞中的表达水平较高;但培养1d后,几乎检测不到HTm4基因的表达;培养3～14d,该基因的表达回升并持续维持在高水平。而HTm4S基因的表达在第7d达最高水平,其余时间基本呈稳定表达。转染HTm4和HTm4S,亚细胞定位结果显示HTm4主要定位于核周围,而HTm4S则定位于整个胞浆,由此可能导致它们功能的区别。以上结果提示,长期培养体现出FRIL在维持造血干／祖细胞多能性上的优势;细胞周期调控基因HTm4及其新剪接子参与了FRIL体外长期维持脐血造血干／祖细胞处于静息状态的过程。     

Neonatal imprinting and hepatic cytochrome P-450 II. Partial purification of a sex-dependent and neonatally imprinted form(s) of cytochrome P-450

A new form of cytochrome P-450 was partially purified from hepatic microsomes of neonatally imprinted rats (adult male and adult male castrated at four weeks of age). This new form of cytochrome P-450 appears to have an apparent molecular weight of approximately 50,000 daltons as judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis. It appears that this form of cytochrome P-450 is either absent or present in low concentrations in cytochrome P-450 preparations isolated from neonatally nonimprinted rats (adult female and adult male castrated at birth). Reconstitution of testosterone hydroxylase and benzphetamine N-demethylase activities of this partially purified cytochrome P-450 revealed that the presence of testosterone 16α-hydroxylase activity, an imprintable microsomal enzyme, was in parallel with the imprinting status of the animals; a significantly higher activity was detected in the neonatally imprinted than that of the nonimprinted animals. This was in contrast to the nonimprintable benzphetamine N-demethylase, testosterone 7α-and 6β-hydroxylase activities which exhibited no correlation with the imprinting status of the animals. We have prepared antisera from rabbits using the partially purified cytochrome P-450 preparations from adult male rats as antigens. These antisera inhibited microsomal testosterone 16α- and 7α-hydroxylase activities in a concentration-dependent manner, without impairing 6β-hydroxylase activity. These data suggest that the partially purified cytochrome P-450 from adult male rats consists of both imprintable (16α-) and nonimprintable (7α-) testosterone hydroxylase activities. The antisera formed immunoprecipitant lines in the Ouchterlony double diffusion plates with partially purified cytochrome P-450 from both neonatally imprinted and nonimprinted adult rats. The immunoprecipitant lines, as stained by coomassie blue, suggest the homology of the cytochrome P-450 preparations from neonatally imprinted and nonimprinted rats. Immunoabsorption of the antisera against neonatally nonimprinted, partially purified cytochrome P-450 completely removed the immunoprecipitant lines without appreciably impairing the inhibitory effects of antisera on the microsomal testosterone 16α-and 7α-hydroxylase activities. In contrast, immunoabsorption of the antisera against partially purified cytochrome P-450 from adult male rats (imprinted) abolished completely both the immunoprecipitant lines and the inhibition on microsomal testosterone hydroxylation reaction (16α and 7α). The inhibitory actin of antisera on testosterone hydroxyulation was also abolished upon boiling the antisera at 100°C for 5 minutes. The biochemical and immunochemical data in this study suggest that the neonatally imprintable form or forms of hepatic microsomal cytochrome P-450 accounts for a small fraction of the bulk of total cytochrome P-450. However, the existence of this form of cytochrome P-450 is regulated by gonadal hormones during the neonatal period and accounts for the major imprintable sex difference in drug and steroid metabolism in adulthood.     

Multiple strategies to improve sensitivity,speed and robustness of isothermal nucleic acid amplification for rapid pathogen detection

Background  

In the past decades the rapid growth of molecular diagnostics (based on either traditional PCR or isothermal amplification technologies) meet the demand for fast and accurate testing. Although isothermal amplification technologies have the advantages of low cost requirements for instruments, the further improvement on sensitivity, speed and robustness is a prerequisite for the applications in rapid pathogen detection, especially at point-of-care diagnostics. Here, we describe and explore several strategies to improve one of the isothermal technologies, helicase-dependent amplification (HDA).     

牙形石Histiodella Hass 四个种的讨论及地层对比意义

作者描述了产自塔里木盆地的HistiodellaHass的四个种,并探讨了它们与中国北方和北美中大陆暖水型牙形石及中国南方和北大西洋冷水型牙形石的对比关系。认为Histiodellasinuosa带对比北美中大陆牙形石动物群3的下部、中国南方Amorphognathusvariabilis带下部和中国北方Aurilobodusleptosomatus-Loxodusdissectus带下部。Histiodellaholodentata带下部相当于中国北方Aurilobodusleptosomatus-Loxodusdissectus带中部或中国南方的Amorphognathusvariabilis带至Eoplacognathussuecicus带的下部。Histiodellakristinae带对比北大西洋区的Eoplacognathussuecicus带中上部和中国北方的Tangshanodustangshanensis带。Histiodellabellburnensis带对比北大西洋区的Eoplacognathussuecicus带的上部到Eoplacognathusfoliaceus带下部。HistiodellaHass与北大西洋区和中国北方区典型牙形石的共同出现可以作为一个桥梁,建立起北美中大陆、北大西洋、塔里木盆地及华北地区牙形石的对比关系。     

Using pseudo-amino acid composition and support vector machine to predict protein structural class

As a result of genome and other sequencing projects, the gap between the number of known protein sequences and the number of known protein structural classes is widening rapidly. In order to narrow this gap, it is vitally important to develop a computational prediction method for fast and accurately determining the protein structural class. In this paper, a novel predictor is developed for predicting protein structural class. It is featured by employing a support vector machine learning system and using a different pseudo-amino acid composition (PseAA), which was introduced to, to some extent, take into account the sequence-order effects to represent protein samples. As a demonstration, the jackknife cross-validation test was performed on a working dataset that contains 204 non-homologous proteins. The predicted results are very encouraging, indicating that the current predictor featured with the PseAA may play an important complementary role to the elegant covariant discriminant predictor and other existing algorithms.     

植物基因打靶技术及其应用

本文对植物基因打靶技术的原理、操作程序、打靶效率的影响因素及其在植物中的应用现状进行了综述,并就如何有效的提高打靶效率提出了建议,同时对该技术在植物学研究领域中的应用前景进行了展望。     

HILI Inhibits TGF-β Signaling by Interacting with Hsp90 and Promoting TβR Degradation

PIWIL2, called HILI in humans, is a member of the PIWI subfamily. This subfamily has highly conserved PAZ and Piwi domains and is implicated in several critical functions, including embryonic development, stem-cell self-renewal, RNA silencing, and translational control. However, the underlying molecular mechanism remains largely unknown. Transforming growth factor-β (TGF-β) is a secreted multifunctional protein that controls several developmental processes and the pathogenesis of many diseases. TGF-β signaling is activated by phosphorylation of transmembrane serine/threonine kinase receptors, TGF-β type II (TβRII), and type I (TβRI), which are stabilized by Hsp90 via specific interactions with this molecular chaperone. Here, we present evidence that HILI suppresses TGF-β signaling by physically associating with Hsp90 in human embryonic kidney cells (HEK-293). Our research shows that HILI mediates the loss of TGF-β-induced Smad2/3 phosphorylation. We also demonstrate that HILI interacts with Hsp90 to prevent formation of Hsp90-TβR heteromeric complexes, and improves ubiquitination and degradation of TβRs dependent on the ubiquitin E3 ligase Smurf2. This work reveals a critical negative regulation level of TGF-β signaling mediated by HILI (human PIWIL2) by its ability to interact with Hsp90 and promote TβR degradation.     

黄腐酸和甜菜碱预处理对干旱胁迫下平邑甜茶生理特性及光合的影响

该研究以平邑甜茶[Malus hupehensis(Pamp.)Rehd.]2年生实生苗为材料,通过盆栽试验于干旱处理前3d分别连续喷施黄腐酸(FA)、甜菜碱(GB)和复配(FA+GB),并以清水为对照(CK)进行预处理,比较分析不同预处理对干旱胁迫下平邑甜茶的生理及光合特性变化,探讨FA和GB对平邑甜茶的抗旱生理机制。结果显示:(1)与对照相比,FA、GB和FA+GB预处理均能够显著提高平邑甜茶叶片相对含水量,且FA的保水性效果最佳。(2)3种预处理均可显著促进干旱胁迫下叶片可溶性蛋白、可溶性糖和脯氨酸含量增加,且FA+GB预处理后在干旱胁迫下叶片可溶性糖和脯氨酸累积量显著高于单施FA或GB。(3)3种预处理均可显著提高干旱胁迫下平邑甜茶幼苗的SOD、POD、CAT活性,并显著降低MDA的积累速度及其累积量,且以FA+GB预处理的MDA含量最低、抗氧化酶活性最高。(4)GB和FA+GB预处理下平邑甜茶的净光合速率、瞬时水分利用率显著高于CK和FA,且FA+GB处理下改善光合特性的效果最佳,GB次之。研究表明,单独喷施黄腐酸和甜菜碱及两者配施预处理均能够增加干旱胁迫下平邑甜茶的渗透调节物质和相对含水量,提高叶片的保水性,调节抗氧化物酶活性,降低丙二醛含量,增加细胞膜稳定性,改善光合性能,进而提高平邑甜茶的抗旱能力,且以复配喷施(FA+GB)预处理的效果最好。