右美托咪定对脑缺血/再灌注大鼠海马兴奋性氨基酸含量及NMDA受体亚单位NR1表达的影响

目的：观察右美托咪定预处理对全脑缺血/再灌注大鼠海马细胞外谷氨酸（Glu）、天门冬氨酸（Asp）含量及N-甲基-D-天冬氨酸（NMDA）受体1（NR1）表达的影响,探讨右美托咪定脑保护作用及其神经递质机制。方法：雄性Wistar大鼠54只,随机分为3组（n=18）：假手术组、脑缺血/再灌注组和右美托咪定预处理组。用四血管闭塞法建立大鼠全脑缺血模型。收集清醒、缺血15 min及再灌注0~1 h微透析标本。于全脑缺血15 min再灌注1 h后,迅速断头取脑,采用免疫组化法和蛋白免疫印迹法检测海马NMDA受体NR1亚单位的表达情况。结果：与脑缺血/再灌注组相应时点比较,右美托咪定预处理组大鼠海马微透析液中Glu、Asp含量明显降低（P<0.05, 0.01）;免疫组化和Western-blot法检测显示右美托咪定预处理组大鼠海马组织NMDA受体亚单位NR1表达明显受抑制（P<0.05, 0.01）。结论：右美托咪定预处理不仅减少脑缺血/再灌注时兴奋性氨基酸释放,还能抑制NMDA受体亚单位NR1的高表达而产生脑保护作用。     

异位（过）表达PGRN抑制IL-1β引起的髓核细胞损伤

炎症因子IL-1β是引起髓核细胞功能异常的关键因素之一。颗粒体蛋白原（progranulin,PGRN）是一种多功能生长因子,在组织修复、炎症反应等过程中发挥重要作用,但其在髓核细胞中的作用尚不清楚。本研究以IL-1β诱导的髓核细胞炎性损伤模型为研究对象,探讨PGRN对IL 1β诱导的髓核细胞损伤的保护作用及其机制。基因转染结合MTT方法证明,与IL-1β处理的细胞比较,过表达PGRN可逆转IL-1β引起的原代培养的髓核细胞生长抑制,促进细胞增殖。TUNEL技术和流式细胞分析显示,PGRN抑制IL-1β诱导的髓核细胞凋亡。Western印迹和RT-qPCR方法揭示,与IL-1β处理的细胞相比,过表达PGRN显著上调聚蛋白聚糖（aggrecan）和II型胶原（collagen type II）的蛋白质表达,但下调基质金属蛋白酶-13（MMP-13）、分解素-金属蛋白酶ADAMTS-5的表达,同时抑制IL-1β诱导的炎性因子IL-6、IL-8和TNF-α的表达,说明PGRN可缓解IL-1β引起的炎性反应,并减少细胞外基质（ECM）相关蛋白质的降解。此外,过表达PGRN还可降低p65、p-IkB a和β-catenin的蛋白质表达水平,提示PGRN可抑制IL-1β下游TNF-α介导的NF-κB信号途径及β-catenin途径。总之,上述结果提示,过表达PGRN可通过抑制IL-1β诱导的炎性反应、髓核细胞凋亡及基质代谢紊乱,缓解IL-1β诱导的髓核细胞损伤;PGRN的这种抗炎、抗基质降解作用可能与PGRN参与调控NF-κB和β-catenin信号途径有关。     

MiR-144通过靶向抑制GRK5表达促进脊髓星形胶质细胞活化

已知miR-144与细胞活化和增殖有关,然而其具体分子机制尚不明确。本研究发现miR-144通过靶向GRK5促进脊髓星形胶质细胞的活化。运用real-time PCR检测脊髓损伤和正常大鼠的脊髓组织及其脊髓星形胶质细胞中miR-144的表达,发现与正常的组织和细胞相比,miR-144在脊髓损伤组织和星形胶质细胞中的表达水平显著降低;Western印迹检测到脊髓损伤大鼠的星形胶质细胞中GFAP蛋白的表达显著低于正常大鼠,而GRK5蛋白的表达高于正常大鼠;MTT分析结果显示转染miR-144可显著提高脊髓损伤大鼠的星形胶质细胞活性,但对细胞增殖无明显作用;酶活性试剂盒分析发现miR-144显著提高了SOD和GSH活性;生物学信息分析和萤光素酶报告基因检测结果显示miR-144能靶向结合GRK5,并下调GRK5的表达;MiR-144 mimic转染或miR-144 mimic与pcDNA-GRK5共转染脊髓损伤的星形胶质细胞,发现miR-144转染能通过激活NF-κB通路消除pcDNA-GRK5引起的细胞活化抑制。综上所述,miR-144通过靶定结合癌基因GRK5来促进脊髓星形胶质细胞细胞的活化。     

Potential advantages of acute kidney injury management by mesenchymal stem cells

Mesenchymal stem cells are currently considered as a promising tool for therapeutic application in acute kidney injury (AKI) management. AKI is characterized by acute tubular injury with rapid loss of renal function. After AKI, inflammation, oxidative stress and excessive deposition of extracellular matrix are the molecular events that ultimately cause the end-stage renal disease. Despite numerous improvement of supportive therapy, the mortality and morbidity among patients remain high. Therefore, exploring novel therapeutic options to treat AKI is mandatory. Numerous evidence in animal models has demonstrated the capability of mesenchymal stem cells (MSCs) to restore kidney function after induced kidney injury. After infusion, MSCs engraft in the injured tissue and release soluble factors and microvesicles that promote cell survival and tissue repairing. Indeed, the main mechanism of action of MSCs in tissue regeneration is the paracrine/endocrine secretion of bioactive molecules. MSCs can be isolated from several tissues, including bone marrow, adipose tissue, and blood cord; pre-treatment procedures to improve MSCs homing and their paracrine function have been also described. This review will focus on the application of cell therapy in AKI and it will summarize preclinical studies in animal models and clinical trials currently ongoing about the use of mesenchymal stem cells after AKI.     

A phenological hypothesis on the thermophilous distribution of Pistacia lentiscus L.

In this study we present evidences of the importance of the phenological pattern on the distribution limits of the dioecious Pistacia lentiscus L. This species, though displaying a thermophilous distribution, has been proved to resist low freezing temperatures during winter. We try to explain this apparent paradox by studying the effects of an extreme cold event that occurred in December 2001 on a natural population of P. lentiscus in the NE of the Iberian Peninsula. In previous phenological studies conduced over 4 years, no spring branch damage was noticeable among population individuals. Female individuals of this species present a prolonged phenophase development which extends until early winter with the conclusion of fruit set. Therefore, we hypothesize that female plants would be more affected than male ones in terms of the following: (1) vegetative and reproductive organs survival; and (2) next-year vegetative growth and fruit production. To test this hypothesis, we selected 225 adult individuals (113 females and 112 males) in April 2002, and estimated their crown volume, percentage of frozen branches and reproductive buds, and the amount of 2001 fruits frozen. In June 2002 we evaluated, in the same individuals, the percentage of vegetative buds flushed into shoots and of reproductive buds producing infrutescences. Branch mortality was significantly higher in female plants and females with increased frost-damage displayed a higher amount of frozen fruits. The loss of reproductive buds caused a decrease in 2002 fruit production, while 2002 vegetative growth was unaffected by the degree of frost damage. These results verify most of the predictions of our hypothesis. Moreover, they suggest that the limited distribution of P. lentiscus in the cold areas of Mediterranean climate could be more determined by the long extent of the phenological activity of the crown than by its frost tolerance during winter.     

Paracrine action of HO-1-modified mesenchymal stem cells mediates cardiac protection and functional improvement

The aim has been to determine whether the supernatants of mesenchymal stem cells (MSCs) transfected with adenovirus carrying human heme oxygenase-1 (hHO-1) gene protect cardiomyocytes from ischemic injury. We have found that hHO-1 infected MSCs (hHO-1-MSCs) increased expression of hHO-1 protein. Apoptosis of cultured hHO-1-MSCs exposed to hypoxia was suppressed. Several cytokines, including HGF, bFGF, TGF-beta, VEGF and IL-1beta, were produced by hHO-1-MSCs, some being significantly enhanced under hypoxia stimulation. Meanwhile, those cytokines reduced caspase-3 level and activity in cultured adult rat ventricular cardiomyocytes (ARVCs) exposed to hypoxia. Supernatants obtained from hHO-1-MSCs improved left ventricular function, limited myocardial infarct size, increased microvessel density, and inhibited apoptosis of cardiomyocytes in rat myocardial infarction. It can be concluded hHO-1-modified MSCs prevent myocardial cell injury via secretion of paracrine-acting mediators.     

Prokaryotic microbiota of recycled paper mills with low or zero effluent

The tendency in the paper industry is to close all water loops to save water. This leads to higher process temperatures and may increase the colloidal and dissolved material in the process circulation. Increase of nutrients in the water circuits may favor microbiological growth and fouling. In this paper the chemical and microbial compositions of water circuits and deposits were studied of two closed cycled paper/board mills, one mill totally closed (0 m(3) waste water t(-1)), and the other low discharging (about 4 m(3 )t(-1)). The zero discharge mill accumulated high amounts (>10 g C L(-1)) of organic carbon in the circulation waters, about 40% of which composed of volatile acids (lactic, acetic, propionic and butyric acid). Water contents of sulfate, chloride, sodium and calcium increased to >1 g L(-1) of each. q-PCR targeted on 16S rRNA genes indicated that the bacteria in water circuits were mainly viable cells. In both mills anaerobic growth (10(6)-10(8) CFU mL(-1)) equalled or exceeded aerobic growth, with odor problem but no actual slime problem. The major part (40%) of all identifiable bacterial sequences were closest but yet distant (<96%) to Enterococcus cecorum and in the 4 m(3 )t(-1) discharging mill also Bacillus thermoamylovorans and Bacillus coagulans. Slimes and deposits from the mills contained high amounts, >/=10(8) g(-1), of archaean, but only the genus Methanothrix was identifiable from the cloned sequences. The findings indicate that closing the water circuits strongly limited diversity of the microbiota but allowed efficient mineralization of the dissolved and suspended matter.     

一氧化氮对番茄种子抗吸胀冷害的影响

以番茄毛粉802种子为材料,通过对比实验,测定分析各处理种子的萌发率及第4天的平均根长、萌发指数、活力指数,以及相对电导率(REC)、丙二醛(MDA)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)含量的变化,以探讨NO对番茄种子吸胀冷害的抵抗作用及其机理.结果显示:(1)外源NO可显著提高番茄种子经12 h吸胀冷害处理后的萌发率、平均根长、萌发指数和活力指数,并显著降低吸胀冷害下REC和MDA含量,同时显著提高SOD和CAT的含量.(2)NO所提高的吸胀冷害处理后种子的SOD和CAT活性不能被RNA合成抑制剂放线菌素D和蛋白质合成抑制剂环己酰亚胺抑制.结果表明,NO可提高番茄种子抵抗吸胀冷害的能力,而且与NO激活了抗氧化系统有关,但NO不是通过促进抗氧化酶的合成来提高其活性.     

Consequences of injuries on survival and reproduction of common bottlenose dolphins (Tursiops truncatus) along the west coast of Florida

Accurate identification of human‐induced injuries that lead to death or interfere with reproduction is important for marine mammal management, as deaths exceeding established limits can lead to restrictions on fisheries or vessel operations. The fates of cetaceans last seen swimming with attached gear, particularly in pelagic fisheries, or with vessel strike lacerations, have been difficult to predict. Survival and reproduction data from long‐term research on resident common bottlenose dolphins near Sarasota, Florida were examined relative to consequences of fishing gear ingestion, line entanglements, vessel strikes, and amputations of unknown origins. Fishing hooks embedded in the throat, goosebeak, or esophagus, or line wrapped around the goosebeak, generally lead to death. Multiple, constrictive line wraps around fin insertions can lead to amputation, blood loss, impaired mobility, or infection. Dolphins with ingested gear or severe entanglements may swim away with the gear, but likely die later. Propeller injuries involving only soft tissue were often survivable. Some dolphins survived amputations of the distal ends of fins, and continued to reproduce. As a precautionary approach, dolphins with ingested gear or severe constrictive entanglements should be considered mortalities, but extrapolations of findings from coastal bottlenose dolphins to other cetaceans and different gear must be done with caution.