HPLC-MS/MS法研究人尿中甲磺酸桂哌齐特的主要代谢产物

目的研究甲磺酸桂哌齐特在人尿中的主要代谢产物。方法选择健康受试者12名,连续5 d单剂量静脉滴注甲磺酸桂哌齐特250 mg,分段收集0²4 h尿样,经蛋白沉淀纯化后,直接采用液相色谱-串联质谱法(HPLC-MS/MS)对尿样进行测定。结果在尿样中鉴定出5个主要代谢物,分别为3-O-去甲基-桂哌齐特-葡萄糖醛酸化物(M1)、3-O-去甲基-桂哌齐特-磺酸化物(M2)、桂哌齐特-吡咯烷衍生物(M3)、桂哌齐特-吡咯烷羧基化物(M4)和3-O-去甲基-桂哌齐特(M5)。结论该方法灵敏、选择性高,为深入研究甲磺酸桂哌齐特在人体内的代谢规律提供了可靠的方法。     

桂哌齐特拮抗氧化低密度脂蛋白对人脐静脉内皮细胞的损伤

目的研究桂哌齐特对氧化低密度脂蛋白(ox-LDL)损伤人脐静脉内皮细胞系ECV304的保护作用。方法传代培养ECV304细胞,随机分为空白对照组、模型组、桂哌齐特(200mg·L~(-1)和400mg·L~(-1))组,与ox-LDL共同孵育24h制模,观察培养上清液中一氧化氮(NO)与丙二醛(MDA)含量的变化,测定Fura-2/AM负载后的内皮细胞的游离静息钙浓度([Ca~(2+)]_i)。结果 ox-LDL诱导的模型组上清液中NO的浓度明显降低,MDA含量显著升高(均P<0.01)。桂哌齐特组NO浓度增加(P<0.01),MDA含量降低均(P<0.05)。与空白对照组相比,模型组的[Ca~(2+)]_i显著升高,桂哌齐特组能降低损伤导致的[Ca~(2+)]_i升高(均P<0.01)。结论桂哌齐特能拮抗ox-LDL对人脐静脉内皮细胞的损伤作用。     

电针对脑缺血-再灌注损伤大鼠脑组织SOD和IL-1β含量的影响

目的观察电针对脑缺血-再灌注(I-R)损伤后大鼠脑组织超氧化物歧化酶(SOD)和白细胞介素(IL)1β含量的影响。方法雄性SD大鼠40只,随机分为五组:水合氯醛+脑I-R组(A组)、水合氯醛+脑I-R+电针组(B组)、丙泊酚+脑I-R组(C组)、丙泊酚+脑I-R+电针组(D组)、假手术组(E组),每组8只。采用四血管阻塞法建立大鼠全脑缺血模型,于再灌注开始后,B组和D组电针百会命门和足三里穴20 min。在缺血后24 h,测定脑组织中的SOD和IL-1β含量变化。结果 I-R 24 h后,与A组比较,B组和D组大鼠脑组织中的SOD含量明显增加(P<0.05),C组IL-1β含量明显降低(P<0.05);与B组比较,C组IL-1β含量明显降低(P<0.01),D组IL-1β含量亦明显降低(P<0.05);与C组比较,D组大鼠脑组织中的SOD含量明显增加(P<0.05);与E组比较,A组大鼠脑组织匀浆中的SOD含量明显降低(P<0.05)。结论电针能使脑I-R损伤大鼠脑组织中的SOD含量明显增加,而丙泊酚可以显著降低IL-1β含量。丙泊酚麻醉下电针可通过影响SODI、L-1β明显减轻大鼠脑I-R损伤。     

马来酸桂哌齐特治疗外伤性蛛网膜下腔出血的疗效观察

目的探讨马来酸桂哌齐特对外伤性蛛网膜下腔出血患者的治疗作用。方法将70例外伤性蛛网膜下腔出血患者随机分为对照组（32例）：给予止血、脱水、营养脑神经药物治疗;治疗组（38例）：除常规治疗外．另加用马来酸桂哌齐特320mg加入到5％葡萄糖液500mL中静脉滴注,每日1次。治疗期间根据患者症状变化、动态监测TCD、动态检查头颅CT．分析马来酸桂哌齐特对外伤性蛛网膜下腔出血的治疗效果。结果马来酸桂哌齐特组血管痉挛的改善程度明显优于对照组,且治疗疗程明显短于对照组。结论马来酸桂哌齐特具有较好的防止脑血管痉挛,改善脑循环,减轻脑水肿。保护脑组织的作用,值得临床推广应用。     

马来酸桂哌齐特治疗颅脑损伤术后患者的临床观察

目的研究马来酸桂哌齐特对中、重型颅脑损伤术后病人的疗效.方法中、重型颅脑损伤术后病人50例,随机分为马来酸桂哌齐特治疗组和对照组,各25例.治疗14d后观察GSC评分变化及脑血管痉挛（CVS）发生率.结果治疗14d后两组GSC评分均显著改善（P＜0.05）,马来酸桂哌齐特治疗组GSC评分改善较对照组更显著（P＜0.05）,且CVS发生率显著低于对照组（P＜0.05）.结论马来酸桂哌齐特能有效缓解脑血管痉挛,改善中、重型颅脑损伤术后病人的意识情况.     

二维三七桂利嗪胶囊对大鼠局灶性脑缺血再灌注损伤的保护作用

目的:观察二维三七桂利嗪胶囊对大鼠局灶性脑缺血再灌注损伤的影响.方法:SD大鼠分别用292,146和73 mg·kg-1的二维三七桂利嗪胶囊灌胃10 d,线拴法制备大鼠大脑中动脉缺血模型(middle cerebral arteryocclusion,MCAO),缺血3 h后再灌注3 h,分别取脑组织测定脑梗死范围、脑指数、脑含水量并观察脑组织形态结构,取血测定血清乳酸脱氢酶(LDH)和超氧化物歧化酶(S0D)活性.结果:3个剂量的二维三七桂利嗪胶囊能够不同程度地缩小缺血再灌注损伤后脑梗死范围、降低脑指数、脑含水量和血清LDH活性,提高SOD活性并减轻光镜下脑组织缺血性损伤.结论:二维三七桂利嗪胶囊对大鼠局灶性脑缺血再灌注损伤具有保护作用.     

甲磺酸桂哌齐特与马来酸桂哌齐特注射液药动学比较研究

目的比较甲磺酸桂哌齐特与马来酸桂哌齐特注射液的药动学特征。方法 12名健康志愿者,按照2×2交叉试验设计分别静脉滴注甲磺酸桂哌齐特或马来酸桂哌齐特,连续5d,每日1次。给药后于规定时间采血并收集尿液。洗脱期为10d。结果甲磺酸桂哌齐特与马来酸桂哌齐特单次给药后t1/2分别为2.20±0.42和2.26±0.36h;AUC0~t分别为19.329±2.879和18.547±3.727μg·h·mL-1;Cmax分别为5.386±0.809和5.345±1.035μg·mL-1;CL分别为13.02±2.10和13.87±3.29L·h-1;24h内桂哌齐特(CPZ)尿累积排泄率分别为55.30%±3.39%和56.22%±3.61%。连续5d静脉滴注甲磺酸桂哌齐特与马来酸桂哌齐特后t1/2分别为2.18±0.33和2.52±0.50h;AUC0~t分别为20.168±3.178和18.750±1.763μg·h·mL-1;Cmax分别为5.541±0.520和5.426±0.730μg·mL-1;Cmin低于检测限;CL分别为12.52±1.94和13.21±1.17L·h-1;MRT0~t分别为3.53±0.24和3.56±0.20h。CPZ的主要药动学参数和尿累积排泄率在单次与多次给药间、不同酸根间差异均无统计学意义(P>0.05)。结论甲磺酸桂哌齐特与马来酸桂哌齐特在人体内药动学行为相似,连续多次给药,体内无蓄积。     

缺血预处理和缺血后处理对肾缺血-再灌注大鼠一氧化氮系统的影响

目的 探讨肾缺血预处理和缺血后处理对肾缺血-再灌注(I-R)损伤的影响机制.方法 SD大鼠50只均分为假手术组(A组)、I-R(B组)、缺血预处理组(C组)、缺血后处理组(D组)和联合处理组(E组).缺血45 rin再灌注24 h后取肾,检测一氧化氮(NO)含量、一氧化氮合酶(NOS)活性、超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量;HE染色观察肾组织病理变化情况,并进行中性粒细胞(PMN)计数和Paller's评分;TUNEL法观察肾组织细胞凋亡情况.结果 与B组比较,C、D、E组的SCr、BUN显著降低、肾组织MDA含量显著减少、肾组织SOD活性和肾NOS、NO水平显著增加、肾小管上皮细胞阳性凋亡率显著降低(P＜0.01).结论 缺血预处理和缺血后处理均能够减轻肾I-R损伤,可能与其增加NOS在肾脏的表达有关.     

马来酸桂哌齐特对冠心病患者甲襞微循环的影响

目的:研究马来酸桂哌齐特对冠心病患者甲襞微循环的影响。方法:将15位冠心病患者在相同基础治疗基础上分为对照组(7人、未用马来酸桂哌齐特)和治疗组(8人、马来酸桂哌齐特240 mg/d),10 d一疗程,前后观察甲襞微循环状态的变化。结果:对照组和治疗组治疗前甲襞微循环状态均无差别,1疗程后,均有明显改善,而治疗组更进一步改善了微循环状态。结论:小样本研究初步证实了马来酸桂哌齐特对微循环有改善作用。     

甲磺酸桂哌齐特注射液在健康志愿者中的药动学研究

目的研究甲磺酸桂哌齐特注射液在健康志愿者中单次给药的药动学特征。方法 12名健康志愿者分别静脉滴注甲磺酸桂哌齐特62.5、125、250和375 mg,血浆及尿液中桂哌齐特浓度以新建立的HPLC-UV法测定。结果 62.5、125、250和375 mg剂量组的主要药动学参数分别为:Cmax(1.148±0.149)、(2.614±0.410)、(5.625±0.940)、(8.887±1.197)μg·m L~(-1);AUC0~14(3.625±0.549)、(8.553±1.733)、(19.849±3.346)、(33.925±5.377)μg·h·m L~(-1);t1/2Z(2.49±1.14)、(2.39±0.41)、(2.43±0.51)、(2.22±0.27)h;24 h内桂哌齐特尿累积排泄率分别为(54.15±4.24)%、(56.09±5.90)%、(55.32±5.02)%和(56.15±5.32)%。结论甲磺酸桂哌齐特在62.5~375 mg内单次给药符合线性动力学过程,24 h内约有55%以原型经肾脏排除。     

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.     

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.     

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.     

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.