加替沙星-壳聚糖纳米粒胶联羊膜药膜的制备及体外实验研究

目的制备一种新型生物多功能复合材料——加替沙星．壳聚糖纳米粒胶联羊膜药膜，并观察其体外缓释性能及抑菌作用。方法采用离子胶联法制备加替沙星－壳聚糖纳米粒并检测其表征，然后将栽药纳米粒加入到纤维蛋白胶中，混匀后再与羊膜胶联制成加替沙星－壳聚糖纳米粒胶联羊膜药膜，光镜及扫描电镜下观察其形态学特征，高效液相色谱法测定其中加替沙星的浓度，微量肉汤稀释法检测其体外抑菌活性。结果加替沙星纳米粒的粒径为291±33nm，载药量为10．46％，包封率为61．32％。载有纳米药物的纤维蛋白胶与羊膜基底面粘合紧密，呈圆形的加替沙星纳米粒附着在胶原纤维的网状结构上。在体外，加替沙星－壳聚糖纳米粒胶联羊膜药膜可持续释药达15天，对金黄色葡萄球菌和铜绿假单胞菌的最低抑菌浓度分别为0．05μg／ml和0．42μg／ml。结论加替沙星－壳聚糖纳米粒胶联羊膜药膜在体外具有良好的缓释性能及抑茵作用。     

纳米Fe3O4协同微生物对黄褐土中PCB30的降解

研究了纳米Fe3O4协同微生物降解黄褐土中的2,4,6 三氯联苯(PCB30),以PCB30为唯一碳源时降解菌的生长状况,以及微生物接菌量、纳米Fe3O4投加量、PCB30浓度对黄褐土中PCB30降解的影响。PCBs降解菌经1 6S rDNA鉴定为假单胞菌Pseudomonas sp.,与草莓假单胞菌Pseudomonas sp.fragi同源性为75%。环境因素对黄褐土中PCB30降解效果有明显影响。微生物接菌量在0~0.8 mL(1×1 09cfu·mL-1)、纳米Fe3O4投加量在0~1 6.7 g·kg-1、PCB30浓度在0~1 0 mg·kg-1范围内时,PCB30残留率随着微生物接菌量、纳米Fe3O4投加量以及PCB30浓度的增大而降低。当三者都分别达到各自范围的上限时,微生物单一体系、纳米Fe3O4单一体系和纳米Fe3O4/微生物协同体系中PCB30残留率在反应7 d后分别为63.1 8%、43.27%和26.28%;纳米Fe3O4/微生物协同体系的降解效果明显优于微生物和纳米Fe3O4单一体系。     

瑞替普酶治疗急性心肌梗死（AMI）的临床观察

目的对比观察瑞替普酶（reteplase rPA）与尿激酶用于急性心肌梗死（AMI）溶栓治疗的效果。方法从2006年10月到2008年10月，共收治38例AMI患者，随机接受r—PA或尿激酶溶栓治疗，观察溶栓再通率及不良反应发生率。结果溶栓后2h再通率rPA组为89．47％，尿激酶组68．42％（P〈0．01）。结论瑞替普酶治疗AMI再通率高，疗效确切，值得临床推广。     

布什提议增加美国太空预算

美国总统布什近日在向国会提交的2006财政年度政府预算中，提议为美国宇航局2006财政年度拨款164．5亿美元，比2005财年增加2．4％。     

江汉平原主要气候变化特征

根据江汉平原8个气象观测站1961年-2004年的气象资料，对该区气候变化特征进行分析，结果发现，该区年平均气温呈现上升趋势，气温增加倾向率为0．259℃／10a，平均最低气温有明显增加趋势而平均最高气温变化幅度不大；秋、冬两季气温上升明显；1993年是江汉平原气温突变年。该区年降水量呈现上升趋势，降水增加倾向率为42．39mm／10a；降水年际间变化明显，年际问变异系数和极值比分别为0．174和1．97；该区存在明显的3个连丰和2个连枯阶段。该区干燥度指数升高，气候向湿润化发展。该研究结果能够更好的指导该区农业生产和湿地保护，同时为政府部门决策提供气象依据.     

灰色及其改进模型在人口预测中的应用

在灰色GM（1，1）模型和优化的等维递补GM（1，1）模型的基础上，构建了灰色＋BP神经网络组合模型。对2001～2005年我国人口的变化分析后，建立人口总量模型进行预测，利用原始教据建立的灰色＋BP神经网络组合模型预测我国2008年以后五年的人口总量为13．39亿、13．49亿、13．60亿、13．7亿、13．79亿，有逐年上升的趋势。经综合误差分析和后验差检验均为“优秀”，说明该模型具有一定的应用价值。     

克氏螯虾壳聚糖的荧光标记及其抗菌活性的研究

目的建立克氏螯虾壳聚糖的荧光标记方法并研究其抗菌活性。方法利用荧光物质5（6）-羧-四甲基罗丹明-5-马来酰亚胺按不同比例标记克氏螯虾壳聚糖，检测标记后克氏螯虾壳聚糖对大肠埃希菌及金黄色葡萄球菌的抗菌效果，并用激光共聚焦显微镜观察壳聚糖对试验菌的作用靶位。结果克氏螯虾壳聚糖与荧光物质（v／v）9：1混合时，所得的荧光标记克氏螯虾壳聚糖对两种试验菌的抗菌活性保留50％。荧光标记克氏螯虾壳聚糖已进入细菌内部。结论控制荧光标记克氏螯虾壳聚糖抗菌活性基团的比例，不影响其抗菌的活性，可作为一种荧光探针，用于克氏螯虾壳聚糖抗菌作用靶位的研究。     

北京温榆河浮游藻类与水质分析

2006年的监测结果显示，温榆河CODM。BOD5、NH4-N、TP和TN含量大于我国地表水环境质量V类标准的1．3～11．7倍，主要污染物为总氮、氨氯和总磷；五项指标（SS、SD、CODMn,TP、TN）的TSIM值为75．11～119．45，属于富营养型水体。温榆河4个监测断面共检出浮游藻类6门148种，绿藻种类最多（48％），其次是硅藻（20．9％）、蓝藻（18．2％），优势种群均为富营养型水体指示种。温榆河浮游藻类密度大，平均为3179．01×10^4 cells／L，绿藻占优势（75．6％），蓝藻次之（14．6％）。     

河岸缓冲带不同植被配置对铵氮去除效果研究

在沈阳市郊浑河和蒲河两岸开展不同植被配置方式下，开展河岸植被缓冲带对地表径流中铵氮污染消减研究。结果表明：河岸植被缓冲带越宽，水中铵氮浓度越高，其对地表径流中铵氮的消减作用越明显；天然植被比人工植被能更好的去除铵氮污染物；当污染浓度升高时，林草混合配置的植被带对铵氮表现出较好的消减率；人工林草地和天然林草地对铵氮的平均消减率分别为31％和25．9％，最高消减率分别为57．9％和63．3％，最高消减率均发生在7m宽度河岸带上，而人工林地对铵氮的消减效果较差。实验结果能够应用于指导自然河岸带的生态保护与人工河岸植被带的建设。     

胃癌侧群细胞耐药性研究及干细胞相关基因检测

目的研究胃癌侧群（Side Population，sP）细胞对化疗药物5-Fu（氟尿嘧啶）的耐药性及可能机制，并检测干细胞相关基因Nanog、Musashi-1及cD44的表达情况。方法选择人胃癌细胞株sGc-7901，以荧光染料H0echst 33342染色，维拉帕米桔抗对照，应用流式细胞仪分选sP细胞和nonsP细胞。细胞耐药实验比较sP细胞与nonsP细胞对化疗药物5．Fu的耐药性差异；westem_h10t检测ABcG2和bcl-2蛋白表达情况；流式细胞仪分析细胞周期；荧光定量PcR检测两组细胞中干细胞相关基因Nanog、Musashi-1及cD44mRNA的表达差异。结果胃癌细胞株sGc．790l中sP细胞的比例为2．8％，sP细胞对5-Fu的耐药存活率明显高于non-sP细胞（P〈0．05），与nonsP细胞相比，sP细胞高表达耐药蛋白ABcG2和抗凋亡蛋白bcl-2，有更多的细胞处于c0／G1期（P〈0．05），并高表达干细胞相关基因Musashi-1和cD44。结论胃癌sGC_7901细胞株中sP细胞对化疗药物5．Fu的耐药性明显高于nonsP细胞，其耐药机制可能与sP细胞高表达耐药蛋白ABcG2和抗凋亡蛋白bcl-2，有更多细胞处于G0／Gl期有关；Musashi-1和cD44可能是相对特异性的胃癌干细胞标志物。     

Cationic host defence peptides: Innate immune regulatory peptides as a novel approach for treating infections

An increase in antibiotic resistance and the emergence of new pathogens has led to an urgent need for alternative approaches to infection management. Immunomodulatory molecules that do not target the pathogen directly, but rather selectively enhance and/or alter host defence mechanisms, are attractive candidates for therapeutic development. Natural cationic host defence peptides represent lead molecules that boost innate immune responses and selectively modulate pathogen-induced inflammatory responses. This review discusses recent evidence exploring the mechanisms of cationic host defence peptides as innate immune regulators, their role in the interface of innate and adaptive immunity, and their potential application as beneficial therapeutics in overcoming infectious diseases. Received 3 November 2006; received after revision 14 December 2006; accepted 22 January 2007     

Targeted inhibition of Livin resensitizes renal cancer cells towards apoptosis

Cancer cells are typically characterized by apoptosis deficiency. In order to investigate a possible role for the anti-apoptotic livin gene in renal cell cancer (RCC), we analyzed its expression in tumor tissue samples and in RCC-derived cell lines. In addition, we studied the contribution of livin to the apoptotic resistance of RCC cells by RNA interference (RNAi). Livin gene expression was detected in a significant portion of RCC tumor tissue specimens (13/14, 92.9%) and tumor-derived cell lines (12/15, 80.0%). Moreover, targeted inhibition of livin by RNAi markedly sensitized RCC cells towards proapoptotic stimuli, such as UV irradiation or the chemotherapeutic drugs etoposide, 5-fluorouracil, and vinblastine. These effects were specific for livin expressing tumor cells. We conclude that livin can contribute significantly to the apoptosis resistance of RCC cells. Targeted inhibition of livin could represent a novel therapeutic strategy to increase the sensitivity of renal cancers towards pro-apoptotic agents. Received 30 November 2006; received after revision 22 February 2007; accepted 20 March 2007     

Cellular and molecular aspects of drugs of the future: meropenem

Meropenem, first synthesized in the late eighties, has become one of the most important beta-lactam antibiotics of the carbapenem subclass used for the treatment of a variety of life-threatening infections. Due to its unique chemical structure, meropenem is not inactivated by the kidney dehydropeptidase I and the majority of microbial beta-lactamases. Its antimicrobial activity is based on its high affinity for the majority of cell wall-synthesizing enzymes, the so-called penicillin-binding proteins, of Gram-positive and -negative bacteria. However, bacteria have evolved several approaches to resist meropenem: (i) by reducing the affinity of the penicillin-binding proteins for the antibiotics, (ii) by decreasing the permeability of the outer membrane of Gram-negative bacteria, (iii) by using efflux pumps, and (iv) by activating zinc-dependent carbapenemases. Meropenem has a low toxicity profile and, in contrast to imipenem, no central nervous system toxicity.     

Spread of antibiotic resistance with food-borne pathogens

This short review summarizes data on antibiotic resistance profiles of common food-borne pathogens like Salmonella sp., Escherichia coli, Campylobacter sp., Listeria monocytogenes, Clostridium perfringens, Staphylococcus aureus, and coagulase-negative staphylococci. As a flashlight on the literature of the last few years, it provides ample evidence that antibiotic resistance traits have entered the microflora of farm animals and the food produced from them. Molecular analysis of the resistance genes, where available, shows that the food microflora is not separated from its human counterpart and conjugative transfer of resistance genes has been demonstrated in vitro and in a few cases in vivo. For example, for Salmonella typhimurium, resistance towards tetracyclines has increased from zero in 1948 to a 98% level in certain epidemic populations of S. typhimurium DT104 in 1998. The high incidence of food-borne pathogens in raw meat and milk together with a high level of therapeutic, prophylactic and nutritional application of antibiotics in agriculture reveals an antibiotic resistance problem of global dimensions. The resistance problem in human medicine will not be solved if there is a constant influx of resistance genes into the human microflora via the food chain.     

Binding of matrilysin-1 to human epithelial cells promotes its activity

Matrix metalloproteinase-7 (MMP-7, matrilysin- 1) modulates crucial biological events by processing many epithelial cell surface-associated effectors. We addressed MMP-7 interaction with human epithelial cells and its resulting activity. In human endometrium, a model of controlled tissue remodeling, proMMP-7 was diffusely immunolocalized inside epithelial cells, whereas MMP-7 delineated their entire plasma membrane. Endometrial explants preferentially retained active MMP-7, but not proMMP-7. Endometrial epithelial cells and carcinoma cells from various tissues bound active MMP-7. Endometrial carcinoma-derived Ishikawa cells showed high affinity (K_D of ~2.5 nM) and capacity (~260 000 sites per cell) for MMP-7. MMP-7 binding decreased by extracting membrane sterols or interfering with heparan sulfate proteoglycans, and was abrogated by tissue inhibitors of metalloproteinase-2 (TIMP-2) or synthetic MMP inhibitors. Bound MMP-7 not only remained fully active towards a macromolecular substrate but also became resistant to TIMP-2. We conclude that MMP-7-selective targeting to the plasma membrane of epithelial cells promotes its activity by conferring resistance to TIMP-2. A. Berton, C. Selvais: These authors contributed equally to this work. P. J. Courtoy, E. Marbaix, H. Emonard: These authors contributed equally to the supervision of this work. Received 20 September 2006; received after revision 30 November 2006; accepted 18 January 2007     

Antibiotic resistance in microbes

The treatment of infectious disease is compromised by the development of antibiotic-resistant strains of microbial pathogens. A variety of biochemical processes are involved that may keep antibiotics out of the cell, alter the target of the drug, or disable the antibiotic. Studies have shown that resistance determinants arise by either of two genetic mechanisms: mutation and acquisition. Antibiotic resistance genes can be disseminated among bacterial populations by several processes, but principally by conjugation. Thus the overall problem of antibiotic resistance is one of genetic ecology and a better understanding of the contributing parameters is necessary to devise rational approaches to reduce the development and spread of antibiotic resistance and so avoid a critical situation in therapy--a return to a pre-antibiotic era.     

The role of <Emphasis Type="Italic">Bacteroides</Emphasis> conjugative transposons in the dissemination of antibiotic resistance genes

Investigations into the mechanisms of antibiotic resistance gene transfer utilized by Bacteroides species have led to a greater understanding of how bacteria transfer antibiotic resistance genes, and what environmental stimuli promote such horizontal transfer events. Although Bacteroides spp. harbor a variety of transmissible elements that are involved in the dissemination of antibiotic resistance genes, it is one particular class of elements, the conjugative transposons, that are responsible for most of the resistance gene transfer in Bacteroides. The potential for Bacteroides conjugative transposons to transfer antibiotic resistance genes extends beyond those genes carried by the conjugative transposon itself, because Bacteroides conjugative transposons are able to mobilize coresident plasmids in trans and in cis, and also stimulate the excision and transfer of unlinked integrated elements called mobilizable transposons. These characteristics of conjugative transposons alone have significant implications for the ecology and spread of antibiotic resistance genes, and in terms of biotechnology. A novel feature of the most widespread family of Bacteroides conjugative transposons, the CTnDOT/ERL family, is that their transfer is stimulated 100- to 1000-fold by low concentrations of tetracycline. This is significant because the use of antibiotics not only selects for resistant Bacteroides strains, but also stimulates their transfer. Other Bacteroides conjugative transposons do not require any induction to stimulate transfer, and hence appear to transfer constitutively. The constitutively transferring elements characterized so far appear to have a broader host range than the CTnDOT/ERL family of conjugative transposons, and the prevalence of these elements is on the increase. Since these constitutively transferring elements do not require induction by antibiotics to stimulate transfer, they have the potential to become as pervasive as the CTnDOT/ERL family of conjugative transposons.     

Melatonin biosynthesis in the thymus of humans and rats

Melatonin is an indoleamine widely distributed in the evolution that shows a great functional versatility, playing an important role as a transmitter of photoperiodic information and exhibiting antioxidant, oncostatic, anti-aging and immunomodulatory properties. In vertebrates, this molecule is produced by the pineal gland and other extrapineal sites. The present study was carried out to investigate the presence of melatonin in thymus and the possibility of an endogenous melatonin synthesis in this organ, in which T cells are matured. In this work, we demonstrate in humans and rats that thymus contains melatonin, expresses the mRNAs encoding N-acetyltransferase and hydroxyindol-O-methyltransferase, the two key enzymes of the melatonin synthesis, and has this biosynthetic machinery activated. In addition, rat thymocytes cultured for 24 h exhibited high levels of melatonin. The results presented here suggest that human and rat thymuses are able to synthesize melatonin, which could have intracrine, autocrine and paracrine functions. Received 30 September 2006; received after revision 30 December 2006; accepted 15 February 2007     

Accessibility of a critical prion protein region involved in strain recognition and its implications for the early detection of prions

Human prion diseases are characterized by the accumulation in the brain of proteinase K (PK)-resistant prion protein designated PrP27 – 30 detectable by the 3F4 antibody against human PrP109 – 112. We recently identified a new PK-resistant PrP species, designated PrP^*20, in uninfected human and animal brains. It was preferentially detected with the 1E4 antibody against human PrP 97 – 108 but not with the anti-PrP 3F4 antibody, although the 3F4 epitope is adjacent to the 1E4 epitope in the PrP^*20 molecule. The present study reveals that removal of the N-terminal amino acids up to residue 91 significantly increases accessibility of the 1E4 antibody to PrP of brains and cultured cells. In contrast to cells expressing wild-type PrP, cells expressing pathogenic mutant PrP accumulate not only PrP^*20 but also a small amount of 3F4-detected PK-resistant PrP27 – 30. Remarkably, during the course of human prion disease, a transition from an increase in 1E4-detected PrP^*20 to the occurrence of the 3F4-detected PrP27 – 30 was observed. Our study suggests that an increase in the level of PrP^*20 characterizes the early stages of prion diseases. Received 17 October 2007; received after revision 5 December 2007; accepted 14 December 2007     

Hydra – ancient model with modern outfit

The control of growth and differentiation is a central question not only for developmental biologists but increasingly for medical research as well. The freshwater polyp hydra was one of the first organisms to be used as a model system for the study of this question. It was chosen because of its simple body plan and because it is made up of only seven to eight different cell types. Recent research has shown that despite their simple body plan, cnidarians already exhibit an impressive repertoire of molecular tools which are responsible for the control of growth and differentiation and amongst which peptides appear to play an important role. Received 25 April 2007; received after revision 31 July 2007; accepted 28 August 2007