Ribozyme和DNAzyme的基因治疗实验应用进展

Ribozyme和DNAzyme具有水解mRNA分子的功能,是阻断基因表达和抗病毒的重要工具。近年来,Ribozyme和DNAzyme在临床治疗研究中已经获得了长足进展,有许多成功的实例。比较了Ribozyme和DNAzyme的差异和特点,总结了它们在抗病毒、抗肿瘤、治疗遗传病、治疗神经系统疾病等方面的临床前研究、应用及进展。     

European Settlement and the Making and Unmaking of Aboriginal Identities

This paper explores variation and change in Aboriginal people's connections to places, and place‐related identity, as a function of their differential historical relationship to a town. Among Aboriginal people who have lived for some decades in camps around Katherine, Northern Territory, descendants of those who appear to have the most clearly discernable long‐term relationship with the area in the vicinity of the town do not relate to places, nor conceptualise them, in stereotypically ‘traditional’ terms. Their relationships to town and nearby places tend to be of an ideologically unelaborated, homely sort. Kinds of territorial relationships their antecedents can be shown to have had to the area have undergone dissolution. The paper seeks to develop discussion of such variation and the historical and sociological processes involved. The Katherine case brings the social and historical significance of ‘towns’ as sites of Aboriginal/non‐Aboriginal interrelationship into focus, and also requires a critical view of notions of ‘group’ that have tended to dominate recent public process and understanding in Australia.     

植物几丁质酶的结构与功能、分类及进化

近年来对几丁质酶的研究越来越深入,资料也愈来愈多,有的植物几丁质酶除具有几丁质酶活性,还具有其它的活性,典型的几丁质酶由－N－端信号区,催化区和C－端延伸区组成,有的还有几丁质结合域,各项能域具有各自的功能,对植物几丁质酶的分类已经过多次改进,目前公认的分成4组9个亚组,有证据表明植物几丁质酶在进化过程中有遗传转座现象,但具有进化过程还有待进一步确证,对几丁质酶与其它一些蛋白的关系的了解有助于理解几丁质酶的起源和进化,由于几丁质酶具有独特的抗真菌特性,因而几丁质酶基因成为目前抗真菌基因工程研究的热之一。     

Clinical aspects of gastric cancer and Helicobacter pylori--screening, prevention, and treatment

Gastric cancer still represents a global health care burden, and in the absence of strategies implemented for early detection, the disease continues to have a dismal prognosis. Patients presenting with clinical manifestations of gastric cancer have limited options for cure. Thus, early detection and prevention play a key role in the fight against gastric cancer. Serologic-based test methods have the potential to detect a subset of patients at high risk of gastric cancer that require a close clinical and endoscopic follow-up. More data have been produced to support Helicobacter pylori eradication as an efficient strategy to prevent gastric cancer. Treatment options for patients with an advanced disease are still limited, but the introduction of new agents opens a more optimistic perspective for the future.     

植物几丁质酶的结构与功能、分类及进化

近年来对几丁质酶的研究越来越深入,资料也愈来愈多。有的植物几丁质酶除具有几丁质酶活性,还具有其它的活性。典型的几丁质酶由N_端信号区、催化区和C_端延伸区组成,有的还有几丁质结合域。各功能域具有各自的功能。对植物几丁质酶的分类已经过多次改进,目前公认的是分成4组9个亚组。有证据表明植物几丁质酶在进化过程中有遗传转座现象,但具体进化过程还有待进一步确证。对几丁质酶与其它一些蛋白的关系的了解有助于理解几丁质酶的起源和进化。由于几丁质酶具有独特的抗真菌特性,因而几丁质酶基因成为目前抗真菌基因工程研究的热点之一。     

Tachykinins and the control of prolactin secretion

Tachykinins are present in the pituitary gland and in brain areas involved in the control of the secretion of pituitary hormones. Tachykinins have been demonstrated to stimulate prolactin release acting directly on the anterior pituitary gland. These peptides have also been revealed to be able to act at the hypothalamic level, interacting with neurotransmitters and neuropeptides that have the potential to affect prolactin secretion. Tachykinins seem to act by stimulating or inhibiting the release of the factors that affect prolactin secretion. Among them, tachykinins have been demonstrated to stimulate oxytocin and vasopressin release, which in turn results in prolactin release. Tachykinins also potentiated the response to vasoactive intestinal peptide (VIP) and reinforced the action of glutamate, which in turn result in prolactin release. They have also been shown to interact with serotonin, a neurotransmitter involved in the control of prolactin secretion. In addition, tachykinins have been shown to inhibit GABA release, a neurotransmitter with prolactin-release inhibiting effect. This inhibition may result in an increased prolactin secretion by removal of the GABA inhibition. On the other hand, tachykinins have also been shown to stimulate dopamine release by the hypothalamus, an action that results in an inhibition of prolactin release. Dopamine is a well known inhibitor of prolactin secretion. In conclusion, although tachykinins have been shown to have a predominantly stimulatory effect on prolactin secretion, especially at the pituitary level, under some circumstances they may also exert an inhibitory influence on prolactin release, by stimulating dopamine release at the hypothalamic level.     

Riboregulators in kidney development and function

The discovery of microRNAs has brought in another level of intricacy in gene regulation. These microRNAs are small non-coding RNAs that have dual ability to act as repressors or inducers of gene activity. MicroRNAs have been implicated in a wide spectrum of biological processes and their expressions have been found to be dysregulated in several diseases. Recently, microRNAs have emerged as a new area of interest in renal development and pathology. MicroRNA profilings have revealed a number of microRNAs that are specific to the kidney or restricted to certain regions of the organ suggesting possible exclusive roles therein. Recently, knockout studies have shown that these riboregulators are critical for normal renal growth and functional renal system. Individual microRNAs have also been identified in renal disease models including kidney cancers, diabetic nephropathy and polycystic kidney disease. Several mechanisms of modulating microRNA activity have also been introduced in recent years. Further progress in the understanding of microRNA activity, identification of microRNA signatures in different states as well as advancement of microRNA manipulation techniques will be valuable for kidney research.     

维管束特异表达启动子及其顺式调控元件

综述了维管束特异表达启动子及其顺式调控元件和基元序列调控基因表达的研究进展 ,它们在抗细菌、真菌性维管束病害的作物基因工程中具有广阔的应用前景。     

Lipids of Salmonella typhimurium and Escherichia coli: structure and metabolism

The nature and quantity of the phospholipids of Salmonella typhimurium and Escherichia coli K-12 have been examined. The main classes of phospholipids, phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin have been completely characterized. Four minor compounds have been detected: phosphatidylserine, phosphatidic acid, and two partially characterized lipids. The phospholipid composition of the two organisms is quite similar, the only difference is the absence of one of the minor components and a decreased level of all components in E. coli. A study of the turnover of the phosphate in the phospholipids demonstrated no turnover in phosphatidylethanolamine, a slow turnover in phosphatidylglycerol, and a slow turnover in cardiolipin with, possibly, a transfer of phosphate from phosphatidylglycerol to cardiolipin. The amino acid phenylalanine is shown to become incorporated intact into lipidic compounds which have been partially characterized. Methods for the isolation and separation of lipids have been examined for their utility with these bacteria.     

Microbiological Standards and Handling Codes for Chilled and Frozen Foods. A Review

The usefulness of microbiological standards for frozen foods is now a controversy in the trade and scientific literature. Most reviewers have given arguments both for and against, and have concluded that they should be applied with great caution. Such standards have the advantage of putting questions of safety on a convenient numerical basis. Canadian workers have reported that promulgation of standards has invariably raised the hygienic level of the products controlled.

Bacteriological standards have often been associated with the question of safety to the consumer. Everyone recognizes that food poisoning bacteria are a potential danger in any food. But many have argued that the history of food poisoning outbreaks from frozen foods is excellent and that there is no need for standards; on the other hand, proponents of standards have pointed to the incomplete investigation and reporting of outbreaks, and have argued that there may be more outbreaks than we realize. They have pointed to laboratory studies that have shown grossly mishandled precooked frozen foods to be truly dangerous. Some have proposed that pathogens should be absent from foods; but others have questioned that a microbiological standard can accomplish this end. Some pathogens, such as Salmonella or Staphylococcus have been shown to be so ubiquitous that their presence in some commercial foods is unavoidable. Also, sampling and analytical methods have been described as inadequate to guarantee that pathogens present will be detected. Some have argued that control at the source is a better way—through inspections of the plant operation, by enforcement of handling codes, or by processing procedures such as pasteurization, which would be more certain to result in a pathogen-free food.

A most important part of any of the proposed standards is a “total count” of viable aerobic bacteria. English workers have found that foods causing poisoning outbreaks usually had total viable counts above 10 million per gram. On the other hand, these same workers found Salmonella on meats with very low total viable count. The assumption by many that low total count indicates safety has been shown to be not always true. Furthermore, high counts of nonpathogenic organisms, such as psychrophilic saprophytes would have no public health significance.

The relation between bacterial level and quality is open to less controversy. Some authorities have pointed to bacterial level as a measure of sanitation, adequacy of refrigeration, or speed of handling. Others have indicated that to determine which of these factors caused a high count would be impossible with only a total count on the product as a guide. Some investigators have said a high count affects flavor adversely before actual spoilage is evident, and this may be a factor in competition on today's market. It is well established that initial bacterial level will affect the shelf-life of a chilled product. Methods of analysis are more nearly adequate for counts than for pathogens, but they need improvement, and should be clearly specified as part of any bacteriological standard. Foods with high count could sometimes be brought into compliance merely by storing them for a sufficient period frozen, or by heating them slightly. This has been cited by some authors as a disadvantage of bacteriological standards.

The enterococci and the coliform group (except Escherichia coli) have been shown to be ubiquitous and therefore should not be used alone to indicate fecal contamination. Although E. coli has greater significance, its source should be determined each time it is found.

Various reviewers have expressed the need for caution in the application of standards. The principal precautionary arguments we have found are as follows:

1) A single set of microbiological standards should not be applied to foods as a miscellaneous group, such as “frozen foods” or “precooked foods.”

2) Microbiological standards should be applied first to the more hazardous types of foods on an individual basis, after sufficient data are accumulated on expected bacterial levels, with consideration of variations in composition, processing procedures, and time of frozen storage.

3) When standards are chosen, there should be a definite relation between the standard and the hazard against which it is meant to protect the public.

4) Methods of sampling and analysis should be carefully studied for reliability and reproducibility among laboratories, and chosen methods should be specified in detail as part of the standard.

5) Tolerances should be included in the standard to account for inaccuracies of sampling and analysis.

6) At first, the standard should be applied on a tentative basis to allow for voluntary compliance before becoming a strictly enforced regulation.

7) Microbiological standards will be expensive to enforce.

8) If standards are unwisely chosen they will not stand in courts of law.

     

Water and Ions in Muscles and Model Systems

The nuclear magnetic reasonance (NMR) relaxation times of protons in toad muscle water have been measured at three frequencies: 2.3, 8.9, and 30 MHz. The results are analyzed in terms of a distribution of correlation times, and it is found that only a few percent of the observed protons have mobilities more than two orders of magnitude smaller than normal. Sodium and chloride ion chemical potentials in some hydrated materials with similar proton NMR characteristics to toad muscle have been found to be heightened, but not sufficiently to account for the distribution of sodium ions in muscle.     

Opiate tolerance and dependence: recent findings and synthesis.

Recent studies have led to a greater understanding of the behavioral, cellular, and molecular mechanisms underlying opiate tolerance and physical dependence. Behavioral studies have demonstrated that both direct pharmacological effects and the learning of interactions between drug effects and environmental cues are important in these phenomena. Behavioral studies have also revealed that N-methyl-D-aspartate receptors may play a role in their development (or acquisition). Although in early cellular studies no consistent role was found for opioid receptors or endogenous opioid peptides in opiate tolerance and dependence, recent experiments suggest that beta-endorphin, enkephalin, and dynorphin neurons may indeed have a role. Finally, studies at the molecular level suggest that a functional decoupling of opioid receptors from GTP-binding proteins (G proteins) may be important. In this review, we discuss these disparate findings and present a synthesis that shows how they might together contribute to the phenomena of opiate tolerance and physical dependence.     

Biomimetic Leukocyte Adhesion： A Review of Microfluidic and Computational Approaches and Applications

Leukocyte rolling and adhesion are complex physiological processes that have received a great deal of attention over the past decade. Significant increases in the knowledge base related to how leukocytes adhere in shear flows have occurred as a result of the development of novel experimental and computational techniques. Micro- and nano-fabrication techniques have enabled the development of novel flow devices for studying leukocyte adhesion in simple and complex geometries. Improvements in computer technology have enabled simulations of complex flow processes to be developed. As a result of these advances in knowledge related to leukocyte adhesion, numerous novel devices have been developed that mimic the leukocyte rolling and adhesion process. Examples of these devices include cell separation and enrichment devices and targeted ultrasound contrast agents. Future advances related to leukocyte rolling and adhesion processes hold great promise for advancing our knowledge of disease processes as well as development of novel therapeutic devices.     

The Opisthokonta and the Ecdysozoa may not be clades: stronger support for the grouping of plant and animal than for animal and fungi and stronger support for the Coelomata than Ecdysozoa

In considering the best possible solutions for answering phylogenetic questions from genomic sequences, we have chosen a strategy that we suggest is superior to others that have gone previously. We have ignored multigene families and instead have used single-gene families. This minimizes the inadvertent analysis of paralogs. We have employed strict data controls and have reasoned that if a protein is not capable of recovering the uncontroversial parts of a phylogenetic tree, then why should we use it for the more controversial parts? We have sliced and diced the data in as many ways as possible in order to uncover the signals in that data. Using this strategy, we have tested two controversial hypotheses concerning eukaryotic phylogenetic relationships: the placement of arthropoda and nematodes and the relationships of animals, plants, and fungi. We have constructed phylogenetic trees from 780 single-gene families from 10 completed genomes and amalgamated these into a single supertree. We have also carried out a total evidence analysis on the only universally distributed protein families that can accurately reconstruct the uncontroversial parts of the phylogenetic tree: a total of five families. In doing so, we ignore the majority of single-gene families that are universally distributed as they do not have the appropriate signals to recover the uncontroversial parts of the tree. We have also ignored every protein that has ever been used previously to address this issue, simply because none of them meet our strict criteria. Using these data controls, site stripping, and multiple analyses, 24 out of 26 analyses strongly support the grouping of vertebrates with arthropods (Coelomata hypothesis) and plants with animals. In the other two analyses, the data were ambivalent. The latter finding overturns an 11-year theory of Eukaryotic evolution; the first confirms what has already been said by others. In the light of this new tree, we re-analyze the evolution of intron gain and loss in the rpL14 gene and find that it is much more compatible with the hypothesis presented here than with the Opisthokonta hypothesis.     

Structure and Secretion of the Graptolite Prosicula, and its Application for Biostratigraphical and Evolutionary Studies

Studies on graptolite taxonomy and phylogeny in recent years have placed great emphasis on the proximal development of the rhabdosome, particularly the presence or absence of a virgella and early thecal growth patterns. As the prosicula was the earliest part of the graptolite skeleton to be secreted, it may also reveal fundamental information about evolutionary relationships within the Graptoloidea. The prosiculae from a variety of Ordovician taxa ranging in age from Tremadoc to Caradoc have been examined using a combination of light microscopy, SEM and TEM. Parameters investigated include the overall morphology, transition into the nema, pattern of longitudinal ridges and spiral line. Taxa show a change from early Tremadoc graptoloids which have a low diaphragm, prominent spiral line and lack longitudinal ridges, through late Tremadoc and early Arenig taxa which have longitudinal cortical bandages or spiralled, paired longitudinal ridges, into later Arenig and Llanvirn forms which have simple longitudinal ridges and indistinct spiral line and diaphragm. With additional work at higher stratigraphical levels, graptolite prosiculae may prove to be useful biostratigraphically when more complete material is absent, such as in palynomorph preparations from subsurface studies.     

Prolidase function in proline metabolism and its medical and biotechnological applications

Prolidase is a multifunctional enzyme that possesses the unique ability to degrade imidodipeptides in which a proline or hydroxyproline residue is located at the C-terminal end. Prolidases have been isolated from archaea and bacteria, where they are thought to participate in proline recycling. In mammalian species, prolidases are found in the cytoplasm and function primarily to liberate proline in the final stage of protein catabolism, particularly during the biosynthesis and degradation of collagen. Collagen comprises nearly one-third of the total protein in the body, and it is essential in maintaining tissue structure and integrity. Prolidase deficiency (PD), a rare autosomal recessive disorder in which mutations in the PEPD gene affect prolidase functionality, tends to have serious and sometimes life-threatening clinical symptoms. Recombinant prolidases have many applications and have been investigated not only as a possible treatment for PD, but also as a part of anti-cancer strategies, a component of biodecontamination cocktails and in the dairy industry. This review will serve to discuss the many in vivo functions of procaryotic and eucaryotic prolidases, as well as the most recent advances in therapeutic and biotechnological application of prolidases.     

Evolutionary origins and directed evolution of RNA

In vitro selection experiments show first and foremost that it is possible that functional nucleic acids can arise from random sequence libraries. Indeed, even simple sequence and structural motifs can prove to be robust binding species and catalysts, indicating that it may have been possible to transition from even the earliest self-replicators to a nascent, RNA-catalyzed metabolism. Because of the diversity of aptamers and ribozymes that can be selected, it is possible to construct a 'fossil record' of the evolution of the RNA world, with in vitro selected catalysts filling in as doppelgangers for molecules long gone. In this way a plausible pathway from simple oligonucleotide replicators to genomic polymerases can be imagined, as can a pathway from basal ribozyme activities to the ribosome. Most importantly, though, in vitro selection experiments can give a true and quantitative idea of the likelihood that these scenarios could have played out in the RNA world. Simple binding species and catalysts could have evolved into other structures and functions. As replicating sequences grew longer, new, more complex functions or faster catalytic activities could have been accessed. Some activities may have been isolated in sequence space, but others could have been approached along large, interconnected neutral networks. As the number, type, and length of ribozymes increased, RNA genomes would have evolved and eventually there would have been no area in a fitness landscape that would have been inaccessible. Self-replication would have inexorably led to life.     

Design and synthesis of mono- and multimeric targeted radiopharmaceuticals based on novel cyclen ligands coupled to anti-MUC1 aptamers for the diagnostic imaging and targeted radiotherapy of cancer

Targeted radiopharmaceuticals offer the possibility of improved tumor imaging and radiotherapy, with reduced side effects. A variety of monoclonal antibodies and antibody fragments have previously been successfully radiolabeled and used in diagnostic imaging and targeted radiotherapy of cancer. Many such antibodies have been shown to recognize the well-characterized MUC1 tumor marker and have recently been in clinical trials. Furthermore, a number of chelators have been synthesized and are currently used as radiopharmaceuticals for imaging and therapy. We now report the synthesis of a novel, cyclen-based ligand with a sulfur-containing arm that offers increased stability of the ligand-metal complex. We have coupled this ligand with previously selected aptamers to the MUC1 tumor marker to generate a novel targeted radiopharmaceutical with improved properties. We have tested the complex against known, commercially available chelators such as MAG3 in model breast cancer systems. To improve the pharmacokinetic properties of the aptamer-based targeted radiopharmaceutical, we have generated multi-aptamer complexes around a central chelator. Such multi-aptamer complexes have increased retention of the complex in circulation, without affecting the lack of immunogenicity of the complex or altering its superior tumor penetration properties.     

Pseudocholinesterase activity and E1 phenotypes in Down's syndrome and mental retardation.

Pseudocholinesterase activity and the phenotypes controlled by the E1 locus have been determined in a sample of 307 Down's syndrome patients and 206 patients suffering from nonspecific mental retardation and compared to those in the healthy population. Both groups of patients have an elevated frequency of phenotypes possessing the rate E1f allele. The mentally retarded patients have a higher mean pseudocholinesterase activity than those with Down's syndrome who, in turn, have activity than the healthy controls.     

Obesity and colorectal cancer: epidemiology, mechanisms and candidate genes

There is increasing evidence that dysregulation of energy homeostasis is associated with colorectal carcinogenesis. Epidemiological data have consistently demonstrated a positive relation between increased body size and colorectal malignancy, whereas mechanistic studies have sought to uncover obesity-related carcinogenic pathways. The phenomenon of "insulin resistance" or the impaired ability to normalize plasma glucose levels has formed the core of these pathways, but other mechanisms have also been advanced. Obesity-induced insulin resistance leads to elevated levels of plasma insulin, glucose and fatty acids. Exposure of the colonocyte to heightened concentrations of insulin may induce a mitogenic effect within these cells, whereas exposure to glucose and fatty acids may induce metabolic perturbations, alterations in cell signaling pathways and oxidative stress. The importance of chronic inflammation in the pathogenesis of obesity has recently been highlighted and may represent an additional mechanism linking increased adiposity to colorectal carcinogenesis. This review provides an overview of the epidemiology of body size and colorectal neoplasia and outlines current knowledge of putative mechanisms advanced to explain this relation. Family based studies have shown that the propensity to become obese is heritable, but this is only manifest in conditions of excess energy intake over expenditure. Inheritance of a genetic profile that predisposes to increased body size may also be predictive of colorectal cancer. Genomewide scans, linkage studies and candidate gene investigations have highlighted more than 400 chromosomal regions that may harbor variants that predispose to increased body size. The genetics underlying the pathogenesis of obesity are likely to be complex, but variants in a range of different genes have already been associated with increased body size and insulin resistance. These include genes encoding elements of insulin signaling, adipocyte metabolism and differentiation, and regulation of energy expenditure. A number of investigators have begun to study genetic variants within these pathways in relation to colorectal neoplasia, but at present data remain limited to a handful of studies. These pathways will be discussed with particular reference to genetic polymorphisms that have been associated with obesity and insulin resistance.