尼莫地平对早期脑出血大鼠AQP4mRNA、MMP-9mRNA表达的影响

目的：探讨尼莫地平对早期脑出血周边组织AQP4mRNA、MMP-9mRNA表达及由脑出血引起的继发性脑水肿与AQP4、MMP-9的关系。方法：制做脑出血大鼠模型,分为假手术组,脑出血组及尼莫地平组,分别观察6h、1d、3d、5d、7d等5个时间点出血周边脑组织含水量、AQP4mRNA、MMP-9mRNA。结果：（1）脑出血组3d含水量达到高峰,第7天明显下降,但仍高于正常水平;尼莫地平组3d含水量达高峰,但低于脑出血组。（2）脑出血组AQP4mRNA、MMP-9mRNA的表达逐渐升高,3d达到高峰,7d明显下降,5个时间点明显高于假手术组。（3）尼莫地平组大鼠脑水肿程度、AQP4mRNA、MMP-9mRNA表达逐渐升高,3d达到高峰,明显低于脑出血组。（4）脑出血的水肿程度与AQP4mRNA、MMP-9mRNA呈正相关。结论：（1）AQP4、MMP-9参与了出血性脑水肿损伤的过程。（2）尼莫地平可以抑制AQP4、MMP-9的表达,进而减轻脑水肿的程度。     

黄体酮对颅脑损伤大鼠脑组织含水量及水通道蛋白-4表达的影响

目的:探讨黄体酮对脑外伤模型大鼠脑组织含水量、水通道蛋白-4(AQP-4)表达的影响机制.方法:SD雄性大鼠165只,随机分为假手术组(55只)、模型组(55只)和黄体酮组(55只),用Freeney法造成大鼠脑挫裂伤模型,干湿重法测定脑组织含水量,并采用免疫组化方法检测脑组织AQP-4蛋白的表达.结果:模型组大鼠伤后各时间点伤侧脑组织含水量、伤灶周围AQP-4蛋白表达水平均明显高于假手术组(均P<0.05);黄体酮治疗组各时间点脑组织含水量、AQP-4表达水平均较模型组降低(P<0.05).结论:黄体酮可以明显减轻创伤后脑水肿,其作用可能与抑制AQP-4在损伤脑组织中的表达有关.     

蛭龙活血通瘀胶囊对局灶性脑缺血大鼠脑组织促血管生成素-1及Tie-2表达的影响

目的观察蛭龙活血通瘀胶囊对局灶性脑组织缺血大鼠脑组织缺血区促血管生成素-1（Ang-1）、上皮生长因子样域酪氨酸激酶-2（Tie-2）表达的影响。方法将120只健康雄性成年SD大鼠随机分为6组,即假手术组,模型组,蛭龙活血通瘀胶囊高剂量组、中剂量组、低剂量组,步长脑心通对照组,各20只,另30只备用。以Zealonga法为标准制作大鼠大脑中动脉阻塞（MCAO）模型,假手术组只行正中切口,但不插入线栓。各用药组均在动物苏醒后灌胃,假手术组和模型组给予等体积生理盐水灌胃,连续7 d。术后7 d,采用Zealonga评分标准进行5分制神经功能缺损评分;心脏灌注固定,断头取脑,采用免疫组化方法检测大脑皮层缺血区Ang-1及Tie-2的蛋白表达。结果蛭龙活血通瘀胶囊治疗组大鼠脑组织缺血区Ang-1及Tie-2表达较模型组明显升高。结论蛭龙活血通瘀胶囊可能通过促进脑组织Ang-1及Tie-2蛋白表达,从而促进脑梗死后缺血区微血管新生,减少脑缺血后血脑屏障的渗漏,降低脑水肿。     

新生大鼠缺氧缺血性脑损伤AQP-4表达及依达拉奉治疗效果的研究

目的：探讨新生大鼠缺氧缺血性脑损伤（hypoxic-ischemic brain damage,HIBD）后脑组织水通道蛋白-4（AQP-4）的表达变化及依达拉奉（edaravone）对其治疗效果的影响。方法：将7日龄Wistar大鼠120只随机分为3组：假手术组、缺氧缺血模型组（HIBD组）及依达拉奉治疗组,每组各40只。每组根据处死的时间不同又分5个亚组：6、24h、3、5、7d组,每亚组各8只,各组分别于不同时间点处死动物,测脑组织含水量,并采用免疫组化方法检测AQP-4的表达。结果：AQP-4在缺氧缺血6h即增强,3d达高峰,5～7d逐渐降低（P〈0.01）;治疗组各时间点AQP-4表达水平较HIBD组明显降低（P〈0.01）。结论：AQP-4参与了HIBD脑水肿的发生发展过程,依达拉奉可降低AQP-4的表达,从而减轻脑水肿,这可能是其脑保护作用机制之一。     

脑出血大鼠血肿周围脑组织Mg~（2＋）水平、MDA含量与脑水肿的关系

目的：观察镁离子（Mg2＋）以及丙二醛（MDA）、在大鼠脑出血血肿周围脑组织中的表达。探讨Mg2＋及脂质氧化与脑水肿的关系。为丰富脑出血后脑水肿治疗奠定一定理论基础。方法：采用干湿重法、酶联免疫吸附法,火焰原子吸收法测50只脑出血模型组大鼠和50只假手术对照组大鼠血肿周围脑组织中含水量、Mg2＋以及MDA水平。结果：（1）脑出血组各时间点脑组织Mg2＋含量均低于对照组（P〈0.05）。脑出血组Mg2＋含量3d达最低,5d升高,5d与7d差别不显著。（2）脑出血组各时间点脑组织MDA含量均高于对照组（P〈0.05）。脑出血组MDA水平于3d达高峰,5d降低,5d与7d差别不显著。（3）脑出血组各时间点脑组织含水量均高于对照组（P〈0.05）。脑出血组脑组织含水量3d达高峰,5d下降,5d与7d差别不显著。结论：①大鼠脑出血血肿周围脑组织脂质氧化指标：MDA水平于血肿初期升高,3d达高峰,5d降低,5d与7d差别不显著。②大鼠脑出血血肿周围脑组织Mg2＋水平于血肿初期降低,3d达最低,5d升高,5d与7d差别不显著。③大鼠脑出血血肿周围脑组织Mg2＋水平与MDA及脑水肿程度呈负相关。     

奥拉西坦对新生大鼠缺氧缺血性脑损伤后AQP-4表达的影响

目的:探讨奥拉西坦对新生大鼠缺氧缺血性脑损伤后AQP-4表达及脑水肿的影响.方法:将120只Wistar大鼠采用随机分组的方法分为假手术组,HIBD组和奥拉西坦干预组.除假手术组以外的两组大鼠均需制备缺氧缺血模型.每组再根据标本采集时间的不同随机分为6h组、12h组、24h组、3d组和5d组,每亚组8只.各组给药情况如下:将奥拉西坦用生理盐水稀释成0.3mg/mL,干预组于HIBD后即刻给予腹腔注射奥拉西坦2mg/kg,每24h给药一次,假手术组及HIBD组不做处理.在各时间点取样测定各组大俗的脑组织含水量和AQP-4的表达情况.结果:三组大鼠在各个时间点的脑组织含水量均表现为假手术组最低,HIBD组最高,奥拉西坦干预组居中,与HIBD组相比奥拉西坦干预组在各采样时间点AQP-4免疫阳性细胞的平均灰度值均明显偏高(P＜0.01).结论:奥拉西坦可在一定程度缓解脑水肿,提高AQP-4的表达量,提示其缓解脑水肿的机制可能为通过促进AQP-4蛋白的表达实现.此外本次研究还发现AQP-4表达与脑组织含水量呈正相关.     

依达拉奉对脑缺血-再灌注大鼠的神经保护作用及其机制研究

目的探讨分析采用依达拉奉药物对脑缺血-再灌注大鼠的神经保护作用机制。方法对12只大鼠,随机平分为假手术组、脑缺血再灌注组以及依达拉奉组,分别于再灌注后3 d、7 d,测定与对比丙二醛(MDA)含量、脑组织含水量、水通道蛋白4(AQP-4)的表达水平以及一氧化氮合酶(NOS)活性。结果再灌注后3 d脑缺血再灌注组MDA含量、脑组织含水量、AQP-4的表达水平与NOS活性,均高于假手术组,差异具有统计学意义(均P<0.05),而依达拉奉组与脑缺血再灌注组相比,差异具有统计学意义(均P<0.05);再灌注后7 d,三组在MDA含量、脑组织含水量与NOS活性等指标对比,差异无统计学意义(均P>0.05),而依达拉奉组AQP-4的表达水平显著低于脑缺血再灌注组,差异具有统计学意义(P<0.05)。结论依达拉奉药物能够降低脑缺血-再灌注大鼠NOS活性、MDA含量、脑组织含水量,抑制AQP-4的表达,从而起到减轻脑水肿,保护大鼠神经的作用。     

水通道蛋白4mRNA在大鼠蛛网膜下腔出血后脑水肿中的表达及尼莫地平对其表达的影响

目的：研究大鼠蛛网膜下腔出血后脑组织AQP4mRNA的表达及脑含水量的变化以及应用尼莫地平对AQP4mRNA表达的影响。方法：健康成年大鼠60只,随机分为对照组、假手术组、SAH组、尼莫地平组,建立蛛网膜下腔出血模型,分别在6h、1d、3d、5d、7d个时间点断头取脑,测定脑含水量,并用RT-PCR法检测AQP4mRNA表达。结果：SAH组脑含水量及脑组织内AQP4在6h后即增加,随时相递增,3d达到高峰,较假手术组及尼莫地平组明显增高（P＜0.05）。结论：大鼠蛛网膜下腔出血后脑水肿的程度与AQP4的表达成正比,这表明AQP4在蛛网膜下腔出血性脑水肿的病理生理中起着非常重要的作用。尼莫地平可以下调AQP4mRNA的表达,从而减轻脑水肿的程度。     

尼莫同对大鼠缺血性脑水肿脑组织内AQP4 mRNA表达的影响

目的研究大鼠缺血性脑水肿脑组织内AQP_4 mRNA表达及脑水含量的变化,以及尼莫同的干预机制。方法健康Wistar大鼠75只,随机分为假手术组、手术组、尼莫同组,建立大脑中动脉闭塞(MCAO)模型,分别在6h、1d、3d、5d、7d5个时间点将大鼠麻醉后断头取脑,测定脑水含量,并用RT-PCR法测定AQP_4 mRNA表达。结果手术组大鼠脑水含量及脑组织内AQP_4 mRNA表达在6h后开始升高,3d后达到高峰,较假手术组和尼莫同组明显增高(P<0.05)。结论缺血性脑水肿脑组织内AQP_4 mRNA呈高表达,提示AQP4与脑水肿形成有关,钙离子拮抗剂尼莫同可以降低脑组织内AQP_4 mRNA表达,从而减轻脑水肿程度。     

大鼠脑出血后MMP-2、MMP-9的表达与神经元凋亡关系的实验研究

目的 探讨脑出血后基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)的表达和血肿周围神经细胞凋亡发生情况之间的关系,以明确MMP-2、MMP-9对脑出血后神经细胞发生凋亡的贡献作用.方法 80只雄性Wistar大鼠按照出血后的时闻分成对照组和出血后6 h、12 h、1 d、2 d、3 d、6 d和10 d组,分别应用免疫组化和流式细胞仪方法检测血肿周围脑组织MMP-2、MMP-9的表达和神经细胞凋亡率,同时检测脑组织的含水量并进行脑出血动物的神经缺损评分,并统计这些指标的相关性.结果 MMP-9,细胞凋亡率,神经功能缺损评分和脑组织含水量的动态变化均一致,脑出血后6 h开始升高,2 d达到高蜂,10 d与对照组差异仍有统计学意义(P<0.05).而MMP-2则是脑出血后1 d开始升高,6 d达到高峰,10 d后开始下降,与细胞凋亡率,神经功能缺损评分和脑组织含水量无明显相关性.结论 神经细胞凋亡和脑组织含水量直接与神经功能缺损相关,而MMP-9的高表达则促进神经细胞的凋亡,脑组织含水量的增加.     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.