Estudo da estabilidade de novas substâncias psicoativas (NPS) em amostras de sangue seco em papel (DBS), e sua aplicação em toxicologia forense / Stability of new psychoactive substances (NPS) in dried blood spots (DBS), and its application in forensic toxicology

Resumo: Introdução: A classe de drogas de abuso conhecida como novas substâncias psicoativas ou, ainda, drogas desenhadas (do inglês new psychoactive substances (NPS) ou designer drugs) é composta por substâncias químicas obtidas a partir de alterações estruturais de substâncias ou mistura de substâncias psicoativas já existentes (e ilícitas), a fim de mimetizar e/ou potencializar os efeitos proporcionados por estas, com a vantagem de circundar a legislação antidrogas vigente. Dentro da classe das NPS, os grupos de maior destaque atualmente são das catinonas sintéticas, dos canabinóides sintéticos e dos NBOMes (feniletilaminas N-2-metoxibenzil substituídas). As catinonas sintéticas, vendidas pela internet como "sais de banho" ou bath salts, são substâncias estruturalmente relacionadas a catinona, um alcalóide presente no Catha edulis (Khat), com propriedades estimulantes. A classe dos NBOMes é derivada da classe dos alucinógenos 2C, a partir da adição de um grupo N-2-metoxibenzil na amina primária. Os NBOMes possuem ação agonista nos receptores de serotonina, especialmente do subtipo 5-HT2A, o que confere os efeitos alucinógenos. Objetivos: Desenvolver e validar métodos para identificação e quantificação de algumas das NPS de maior relevância em amostras de manchas de sangue seco em papel (do inglês dried blood spots, DBS) e comparar a estabilidade entre dois tipos de amostras (DBS e sangue total), em três diferentes temperaturas (ambiente, 4 °C e -20 °C) e períodos de armazenamento. Metodologia: As amostras de DBS e sangue total foram analisadas em cromatógrafo líquido acoplado à espectrômetro de massas com analisador de massas triplo quadrupolar (LC-MS/MS), utilizando os métodos primeiramente desenvolvidos e validados para cada classe de NPS (catinonas sintéticas e NBOMes). Resultados: A técnica de DBS apresentou um aumento significativo da estabilidade das NPS em comparação à técnica convencional de armazenamento de sangue total. Para os NBOMes, observou-se que os compostos eram estáveis no DBS por período de tempo de até 6 meses, nas três temperaturas estudadas, enquanto que no sangue total, os analitos tiveram uma queda maior que 20% da concentração em 15 ou 30 dias em temperatura ambiente e 180 dias para 4 °C. Para as catinonas, em temperatura ambiente, observou-se baixa estabilidade nas duas matrizes. Porém, para armazenamento à 4 °C, observou-se uma queda na concentração inicial maior que 20% com 60 e 90 dias para mefedrona e benzedrona em DBS, respetivamente, contra 45 e 25 dias. Catinonas que possuem grupamento metilenodioxi em sua estrutura, como a butilona e a pentilona, foram mais estáveis, independente da matriz, em comparação àquelas com grupamento alquila no anel aromático, como mefedrona e benzedrona. Conclusão: A técnica de DBS se mostrou vantajosa para análises forenses em comparação à técnica convencional de armazenamento de sangue total. Além de facilitar o armazenamento (por ocupar menos espaço), sua extração se torna mais rápida e facilitada, já que envolve menos etapas, além de tornar possível a identificação dos analitos de interesse por um maior período de tempo, podendo facilmente ser aplicada na rotina laboratorial(AU)

Abstract: Introduction: New psychoactive substances (NPS) or designer drugs is a class of drugs of abuse composed of chemical substances obtained from structural alterations of substances or mixture of existing psychoactive substances (and illicit), in order to mimic and/or maximize their effects, with the advantage of circumventing existing anti-drug legislation. Within the NPS class, the most prominent groups currently are synthetic cathinones, synthetic cannabinoids and NBOMes (phenylethylamines N-2-methoxybenzyl substituted). Synthetic cathinones, sold by the internet as "bath salts", are substances structurally related to cathinone, an alkaloid present in Catha edulis (Khat), with stimulant properties. The class of NBOMes is derived from the hallucinogen class 2C, from the addition of an N-2-methoxybenzyl group to the primary amine. NBOMes have agonist action at serotonin receptors, especially the 5-HT2A subtype, which confers the hallucinogenic effects. Objectives: To develop and validate methods to identify and quantify some of the most relevant NPS in dried blood spots (DBS) samples and to compare the stability between two matrixes (DBS and whole blood), at three different temperatures (ambient, 4 °C and -20 °C) and storage periods. Method: DBS and whole blood samples were analyzed using a liquid chromatography tandem mass spectrometer with a triple quadrupole analyzer (LC-MS/MS), using the developed and validated methods for each class of NPS (synthetic cathinones and NBOMes). Results: The DBS technique showed a significant increase in the NPS stability compared to the conventional whole blood storage technique. For the NBOMes, the compounds were found to be stable in DBS for a time period of up to 6 months, at the three temperatures studied, whereas in whole blood, the analytes had a decreased higher than 20% of the initial concentration in 15 or 30 days at room temperature and 180 days at 4 °C. For the cathinones, at room temperature, low stability was observed in both matrixes. However, for storage at 4 °C, the initial concentration decreased more than 20% at 60 and 90 days for mephedrone and benzedrone in DBS, respectively, against 45 and 25 days. Cathinones having methylenedioxy group in their structure, such as butylone and pentylone, were more stable, independent of the matrix, compared to those with alkyl group on the aromatic ring, such as mephedrone and benzedrone. Conclusion: The DBS technique proved to be advantageous for forensic analysis compared to the conventional storage technique. In addition to occupying less storage space, DBS extraction becomes faster and easier, since it involves fewer steps, besides to make possible to identify the analytes of interest for a longer period of time, which can easily be applied in the laboratory routine(AU)

Lateral flow immunoassay for quantitative detection of ractopamine in swine urine / 生物医学与环境科学（英文）

A strip reader based lateral flow immunoassay (LFIA) was established for the rapid and quantitative detection of ractopamine (RAC) in swine urine. The ratio of the optical densities (ODs) of the test line (AT) to that of the control line (AC) was used to effectively minimize interference among strips and sample variations. The linear range for the quantitative detection of RAC was 0.2 ng/mL to 3.5 ng/mL with a median inhibitory concentration (IC50) of 0.59 ± 0.06 ng/mL. The limit of detection (LOD) of the LFIA was 0.13 ng/mL. The intra-assay recovery rates were 92.97%, 97.25%, and 107.41%, whereas the inter-assay rates were 80.07%, 108.17%, and 93.7%, respectively.

Validated chromatographic and spectrophotometric methods for analysis of some amoebicide drugs in their combined pharmaceutical preparation

This work is concerned with development and validation of chromatographic and spectrophotometric methods for analysis of Mebeverine HC1 [MEH], Diloxanide furoate [DF] and Metronidazole [MET] in Dimetrol tablets -spectrophotometric and RP-HPLC methods using UV detection. The developed spectrophotometric methods depend on determination of MEH and DF in the combined dosage form using the successive derivative ratio spectra method which depends on derivatization of the obtained ratio spectra in two steps using methanol as a solvent and measuring MEH at 226.4-232.2 nm [peak to peak] and DF at 260.6-264.8 nm [peak to peak]. While MET concentrations were determined using first derivative [[1]D] at lambda = 327 nm using the same solvent. The chromatographic method depends on HPLC separation on ODS column and elution with a mobile phase consisting water: methanol: triethylamine [25: 75: 0.5, by volume, orthophosphoric acid to pH =4]. Pumping the mobile phase at 0.7 ml min[-1] with UV at 230 nm. Factors affecting the developed methods were studied and optimized, moreover, they have been validated as per ICH guideline and the results demonstrated that the suggested methods are reproducible, reliable and can be applied for routine use with short time of analysis. Statistical analysis of the two developed methods with each other using F and student's-t tests showed no significant difference

Caffeine-induced endothelial cell death and the inhibition of angiogenesis / 대한해부학회지

Numerous studies have shown that adenosine or adenosine agonists can stimulate angiogenesis. However, the effect of caffeine (a known adenosine receptor antagonist) on angiogenesis has not been previously studied. Accordingly, this study was undertaken to examine the effect of caffeine on angiogenesis and to clarify the mechanism involved. Chick chorioallantoic membrane assays were used to investigate the effect of caffeine on angiogenesis and proliferation assays using human umbilical vein endothelial cells (HUVECs), were used to study its effects on specific aspects of angiogenesis. The expressions of caspase-3 and Bcl-2 were examined by western blotting, immunofluorescence staining was used to identify HUVEC morphological changes, and fluorescence activated cell sorting (FACS) and DAPI staining were used to detect HUVEC apoptosis. Caffeine was found to inhibit blood vessel formation dose-dependently and to inhibit the proliferation of HUVECs time- and dose-dependently. FACS analysis and DAPI staining showed that inhibitory effect of caffeine on HUVEC proliferation was the result of apoptosis and the up-regulation of thrombospondin-1 (TSP-1). Furthermore, TSP-1 levels were down-regulated by NECA but were unaffected by CGS21680, indicating that caffeine regulated TSP-1 expression via adenosine A2B receptor. In addition, caffeine up-regulated caspase-3 and down-regulated Bcl-2 at the protein level. These results suggest that the inhibitory effect of caffeine on angiogenesis is associated, at least in part, with its induction of endothelial cell apoptosis, probably mediated by a caspase-3 dependent mechanism.

Phenylethylamines from Browningia candelaris (Cactaceae) / Feniletilaminas de Browningia candelaris (Cactaceae)

The analysis by gas chromatography coupled with mass spectrometry (GC-MS) of the alkaloidal extract of Browningia candelaris (Cactaceae) showed the presence of N-acetyl-3,4-dimethoxyphenylethylamine; N,N-dimethyl-3,4-dimethoxyphenylethylamine; N,N-dimethyl-4-methoxyphenylethylamine; and 4-methoxyamphetamine. The presence of these psychoactive compounds is discussed in terms of their possible ritual use in Andean cultures of Northern Chile.

El análisis por medio de cromatografía de gases acoplada a espectrometría de masas (CG-EM) del extracto alcaloidal de Browningia candelaris (Cactaceae) mostró la presencia de N-acetil-3,4-dimetoxifeniletilamina; N,N-dimetil-3,4-dimetoxifeniletilamina; N,N-dimetil-4-metoxifeniletilamina y 4-metoxianfetamina. La presencia de estos compuestos psicoactivos se discute en términos de su posible utilización en ceremonias mágico-religiosas por culturas andinas del norte de Chile.

Regulation of epithelial sodium channel α-subunit expression by adenosine receptor A₂a in alveolar epithelial cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The amiloride-sensitive epithelial sodium channel α-subunit (α-ENaC) is an important factor for alveolar fluid clearance during acute lung injury. The relationship between adenosine receptor A(2a) (A(2a)AR) expressed in alveolar epithelial cells and α-ENaC is poorly understood. We targeted the A(2a)AR in this study to investigate its role in the expression of α-ENaC and in acute lung injury.</p><p><b>METHODS</b>A549 cells were incubated with different concentrations of A(2a)AR agonist CGS-21680 and with 100 µmol/L CGS-21680 for various times. Rats were treated with lipopolysaccharide (LPS) after CGS-21680 was injected. Animals were sacrificed and tissue was harvested for evaluation of lung injury by analysis of the lung wet-to-dry weight ratio, lung permeability and myeloperoxidase activity. RT-PCR and Western blotting were used to determine the mRNA and protein expression levels of α-ENaC in A549 cells and alveolar type II epithelial cells.</p><p><b>RESULTS</b>Both mRNA and protein levels of α-ENaC were markedly higher from 4 hours to 24 hours after exposure to 100 µmol/L CGS-21680. There were significant changes from 0.1 µmol/L to 100 µmol/L CGS-21680, with a positive correlation between increased concentrations of CGS-21680 and expression of α-ENaC. Treatment with CGS-21680 during LPS induced lung injury protected the lung and promoted α-ENaC expression in the alveolar epithelial cells.</p><p><b>CONCLUSION</b>Activation of A(2a)AR has a protective effect during the lung injury, which may be beneficial to the prognosis of acute lung injury.</p>

The Evaluation of Otilonium Bromide Treatment in Asian Patients With Irritable Bowel Syndrome

BACKGROUND/AIMS: Antispasmodics including otilonium bromide (OB) are recommended to treat irritable bowel syndrome (IBS). However, reports about OB experience in Asia is sparse. The purpose of present study was to provide the efficacy of OB in treating Asian IBS patients. METHODS: Overall, 117 IBS patients meeting Rome II criteria were enrolled in an 8-week, double-blind, active-controlled and single center trial. Randomized participants received either OB 40 mg or mebeverine 100 mg 3 doses daily. The primary endpoints were to evaluate the net changes of abdominal pain/discomfort frequency score (APDFS) and safety profile, while the secondary endpoints were to assess the changes in abdominal pain/discomfort intensity, flatulence, abdominal bloating, satisfied stool frequency etc. RESULTS: Finally, 49 OB and 52 mebeverine subjects were eligible for efficacy analysis. Compared to baselines in per protocol populations, the reduced APDFSs in OB and mebeverine were 0.55 +/- 1.20 (P = 0.011) and 0.37 +/- 1.11 (P = 0.042), respectively, to show similarly reduced scores. The most reported side effects included dry mouth, nausea and dizziness. Besides, the improved APDFSs at 4th week visit, final alleviations in abdominal pain intensity, flatulence, abdominal bloating and satisfied stool frequency with global assessments filled by both patients and investigators were significantly achieved by both treatments, and OB was not inferior to mebeverine in treating these parameters. CONCLUSIONS: In Orientals, OB is as effective as mebeverine for alleviating IBS symptoms in terms of abdominal pain, flatulence, abdominal bloating etc. However, obvious side effects are also observed. A large-scaled trial and post-marketing surveillance are recommended to confirm its efficacy and safety.

Repeated Exposure to beta-phenylethylamine Produces Locomotor Sensitization to Amphetamine, but Not Vice Versa, in the Rat

Repeated administration of amphetamine (AMPH) produces behavioral sensitization, a proposed model for the escalation of drug use characteristic of human addicts. beta-Phenylethylamine (PEA) is an endogenous trace amine found in mammalian brain and resembles AMPH both structurally and behaviorally. Previously, it has been reported that chronic PEA administration produces behavioral sensitization to the challenges of AMPH. However, these data were obtained with very high amount of PEA for a relatively long period of time. Further, the effect of PEA challenge on the expression of behavioral sensitization developed by AMPH pre-exposures has not been tested yet. Thus, we examined in the present experiment the expression of behavioral sensitization with AMPH challenge after a mild chronic PEA treatment. Rats were repeatedly administered with systemic injections of saline, beta-phenylethylamine (PEA) (10 or 50 mg/kg), or amphetamine (AMPH) (1.5 mg/kg). When challenged a week after the last pre-injection, rats pre-exposed to either PEA or AMPH showed behavioral sensitization to AMPH (1.0 mg/kg), while these effects were not observed to PEA (50 mg/kg) itself. These results demonstrate that repeated exposure to PEA produces behavioral sensitization to AMPH challenge, while PEA challenge has no effect on the expression of behavioral sensitization developed by AMPH pre-exposures, suggesting that PEA may play a role in the development of locomotor sensitization to AMPH, but not in the expression of it.

Concentrações plasmáticas de triptamina, tiramina e feniletilamina em eqüinos sob efeitos de sobrecarga de carboidratos e antiinflamatórios não esteroidais / Plasmatic concentrations of tryptamine, tyramine end phenylethylamine in horses under the effect of carbohydrate overload and non-steroid antinflammatory compounds

As concentrações plasmáticas das aminas triptamina (TRP), tyramina (TYR) e pheniletilamina (PEA) foram determinadas por cromatografia gasosa (CG) de 20 eqüinos sob efeito de sobrecarga por carboidratos (SC). Após 36h da SC os animais foram aleatoriamente divididos em quatro grupos (n=5) e receberam a cada 12h por via iv: solução salina 10mL (GC), ketoprofeno 2,2mg/kg (GK), fenilbutazona 4,4mg/kg (GF) e flunixin meglumine 1,1mg/kg (GFM). As concentrações das aminas TYR e PEA variaram de 0,18 a 164,2mg/L, com diferenças nos tempos avaliados, mas não entre os tratamentos (p<0,01). A concentração plasmática de TRP apresentou diferenças entre os tempos e também entre os tratamentos. O GC diferiu dos demais nos momentos 48h e 60h e as concentrações nos grupos GK e GFM foram menores que nos grupos GF e GC às 72h (P= 0,0012). Conclui-se que nas doses utilizadas os antiinflamatórios não esteroidais avaliados não interferem nas concentrações de TYR e PEA. Entretanto, o ketoprofeno e o flunixin meglumine foram efetivos em diminuir a concentração plasmática de TRP.

The concentrations of the bioactives amines tryptamine (TRP), tyramine (TYR) and phenylethylamine (PEA) were determined by gas chromatography in plasma samples of 20 horses submitted to carbohydrate overload. Thirty hours after the overload, the horses were randomly distributed in four groups (n=5) and were submitted to four IV treatments every 12 hours: 10ml of saline (GC), ketoprofen 2.2mg/kg (GK), phenylbutazone 4.4mg/kg (GF), and flunixin meglumine 1.1mg/kg (GFM). Blood samples were collected at various times after the overload (0-72 h). Plasma TYR and PEA concentrations ranged from 0.18 to 164.2mg/L, and differed significantly with time (p<0.01), but did not differ in the treatments. Plasma concentrations of TRP differed between times and treatments. The GC was significantly major than other treatments at 48h and 60h after the overload, and the plasma concentration of TRP in groups GK and GFM was significantly lower than in groups GF and GC at 72 h (p=0.0012). We concluded that the anti-inflammatory drugs evaluated do not interfere in the plasma concentration of TYP and PEA. For TRP, ketoprofen and flunixin meglumine was effective to reduce de plasmatic concentration of this amine.

Different effects of adenosine A2A receptors in the models of traumatic brain injury and peripheral tissue injury / 生理学报

Recently, activation of the adenosine A2A receptors has been shown to exert protection against peripheral tissue injuries but aggravation in the central nervous system (CNS) injuries. To explore the different effects of adenosine A2A receptors and try to perform some new treatment strategies for peripheral tissue and CNS traumas, we constructed the mouse models of skin trauma, skin combined radiation-impaired wound and traumatic brain injury (TBI), respectively. Wild type mice and A2A receptor gene knockout mice were both used in the experiments. In skin trauma and combined radiation-impaired wound models, the time of wound healing was observed, while in TBI model, neurological deficit scores, water content in injured brain and glutamate concentration in cerebral spinal fluid (CSF) were detected at 24 h after TBI. The results showed that in skin trauma and combined radiation-impaired wound models, CGS21680 (an agonist of the A2A receptors) promoted while A2A receptor gene knockout delayed the course of skin wound healing. On the contrary, in TBI model, A2A receptor gene knockout, not CGS21680, showed a protective role by inhibition of glutamate release. These data further indicate that promoting glutamate release may account for the different effects of A2A receptor activation in CNS injury and peripheral tissue injury models. These findings may provide some experimental evidence and a new strategy for clinical treatment of peripheral tissue damages by agonists of A2A receptors, while treatment of CNS injuries by antagonists of A2A receptors.

Spectrophotometric determination of some drugs acting on the GIT via complexation of the drugs hydroxamate with iron [III]

Simple spectrophotometric method for the determination of trimebutine maleate, mebevrine HCL, diloxanide furoate and oxyphenonium bromide by hydroxamic acid formation either in pure or dosage forms is described. The studied drugs react with hydroxyl amine in alkaline medium to form hydroxamic acid derivatives that were complexed with iron [III] in acidic medium to form brownish violet colored complexes having maximum absorbances at 535 nm for mebevrine HCL. trimebutine maleate and diloxanide furoate and at 510 nm for oxyphenonium bromide, Beer's law was obeyed in the concentration ranges of 50-450, 100-700, 40-200 and 100-700 .microg ml[-1] for mebevrine HCL, trimebutine maleate, diloxanide furoate and oxyphenonium bromide, respectively. Molar absorbtivities and Sandell's sensitivities were calculated and presented. The developed procedures were favorably applied for determination of the studied drugs in their pharmaceutical dosage forms. The obtained results were compared satisfactorily with the official and reported methods. There were no significant differences between the proposed, official and reported methods

Nuevos antiarrítmicos en lafibrilación auricular / New antiarrhythmic agents in atrial fibrillation

The convenience to count with a safe and effective pharmacological wealth for atrial fibrillation treatment had conduced, in a way, to a deep depuration of the vast array of antiarrhythmic drugs, keeping only a very restricted number of compounds with a widely proved anti-atrial activity. On the other hand, it had lead to the discovery of the pathophysiological concepts that point to novel therapeutic targets. Within these objectives is that new antiarrhythmic drugs with preferential, even selective, activity on myocardial atrium ion channels had been developed. Among these new antiarrhythmics, dofetilide, and AVE0118, are taken into account. In addition, new possibilities are opened based on the knowledge of the cardioprotective-antiarrhythmic qualities of the opioidergic system.

Assessment of moist exposed burn ointment [MEBO] in the healing of chronic pressure ulcers: a prospective comparative clinical study

Pressure ulcers are a serious health issue causing clinical, financial, and emotional challenges. Treatment modalities that promote wound healing are therefore warranted. To assess the efficacy and safety of MEBO [Julphar Gulf Phannaceutical Industries, UAE] as compared to topical antibiotic ointment [Fucidin, Leo Pharmaceutical, Denmark] in the healing of chronic pressure ulcers. During a 4-year period [Jan 2003-Jan 2007], 45 patients of both sexes with 87 pressure ulcers seen at the Ahmadi Hospital [Kuwait] were categorized into 2 groups, those in group 1 [n=22] received MEBO while those in group 2 [n=23] received Fucidin. Age of the patients ranged between 14 and 102 years with a mean of 69, +/- 7.4 years. Patients had their ulcers prior to study entry for a mean of 10.55 months [range 2-26 months]. Data collected included demographics, nutritional status, underlying predisposing disease and co-morbidities. Ulcer surface area [SA] and healing index [HI] were calculated and compared at two-week intervals for 12 weeks. Patients in both groups had similar demographic and clinical features regarding their age, gender, underlying predisposing factor and comorbidities. Ulcer characteristics were also similar with respect to their number, site, size, duration, depth and presence or absence of sepsis. There was a significant [P<0.05] increase in HI and reduction in ulcer SA on weeks two and four respectively, that was main- tamed through 12 weeks in patients receiving MEBO as compared to those receiving Fucidin. Cumulative patient sample percentage showed that 55.8% of ulcers treated with MEBO had complete healing [HI = 1] by 12 weeks, as opposed to only 20.5% of those treated with Fucidin [P<0.001]. Moreover, none of the patients receiving MEBO had a HI of <50% by 12 weeks as compared to 27.3% of those receiving Fucidin [P<0.001]. Linear regression analysis showed that the change in ulcer size and HI can be attributed to ointment application [r2> 0.4]. No adverse effects of ointment were encountered in either group. Based on the data presented, it may be concluded that in addition to its safety, application of MEBO significantly promotes the healing of chronic pressure ulcers with significant increase in HI of any given ulcer to be expected as early as two weeks following initiation of treatment

Studies on constituents from the fermentation of Alternalia sp / 中国中药杂志

<p><b>OBJECTIVE</b>To study the metabolites of marine fungus Alternalia sp.</p><p><b>METHOD</b>Compounds were separated by column chromatography and their structures were elucidated by means of chemical and spectral analysis.</p><p><b>RESULT</b>Six compounds were isolated from the ethyl acetate and n-butanol extracts of the fermentation of marine fungus Alternalia sp. Their structures were elucidated as p-benzyloxy-phenol ( I ), p-hydroxy phenyl ethylamine( II ), 3-hydroxymethyl-8-hydroxyl-pyrrolopiperazine-2, 5-dione ( III ), 3-isobutyl-6-secbutyl-piperazine-2, 5-dione (IV), 5alpha, 8alpha-epidioxy-ergosta-6, 22-diene-3beta-ol (V), 3beta-hydroxxy-cholesta-5-ene (VI).</p><p><b>CONCLUSION</b>Compounds I , II, III, IV have the activity of inducing morphological deformation of mycelia germinated from conidia of Pyricularia oryzae. Compounds I , II , III were isolated from the genus Alternalia for the first time.</p>

Protective effects of 1-(2,6-dimethylphenoxy)-2-(3,4-dimethoxyphenylethylamino) propane hydrochloride(DDPH) on brain ischemia injury in rats / 药学学报

<p><b>AIM</b>To study the effects of 1-(2,6-dimethylphenoxy)-2-(3,4-dimethoxyphenylethylamino) propane hydrochloride(DDPH) on brain ischemia injury in rats.</p><p><b>METHODS</b>By using the middle cerebral artery occlusion (MCAO) induced by nylon surgical thread inserted through the internal carotid artery into the anterior cerebral artery in rats, the effects of DDPH on neuron defects(ND) and infarct size(IS) were investigated. Using incomplete cerebral ischemia in rats, the effects of DDPH on superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in brain tissue and pathological changes in rats were studied.</p><p><b>RESULTS</b>DDPH at the dose of 10 mg.kg-1 i.p. 30 min before ischemia decreased the ND 3 h after ischemia. The IS declined 24 h after ischemia as well. Meanwhile, DDPH was found to increase SOD activity and reduce the MDA content, as well as mitigate pathological damage, of neuron after brain ischemia in rats.</p><p><b>CONCLUSION</b>DDPH showed protective effects on brain ischemia, probably related to its properties of calcium antagonistic effect and increasing the activity of superoxide dismutases.</p>

Protein kinase A-mediated phosphorylation of HERG potassium channels in a human cell line / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the molecular mechanism of human ether-a-go-go-related gene (HERG) potassium channels regulated by protein kinase A (PKA) in a human cell line.</p><p><b>METHODS</b>HERG channels were stably expressed in human embryonic kidney (HEK) 293 cells, and currents were measured with the patch clamp technique. The direct phosphorylation of HERG channel proteins expressed heterologously in Xenopus laevis oocytes was examined by (32)P labeling and immunoprecipitation with an anti-HERG antibody.</p><p><b>RESULTS</b>Elevation of the intracellular cAMP-concentration by incubation with the adenylate cyclase activator, forskolin (10 micromol/L), and the broad range phosphodiesterase inhibitor, IBMX (100 micromol/L), caused a HERG tail current reduction of 83.2%. In addition, direct application of the membrane permeable cAMP analog, 8-Br-cAMP (500 micromol/L), reduced the tail current amplitude by 29.3%. Intracellular application of the catalytic subunit of protein kinase A (200 U/ml) led to a tail current decrease by 56.9% and shifted the activation curve by 15.4 mV towards more positive potentials. HERG WT proteins showed two phosphorylated bands, an upper band with a molecular mass of approximately 155 kDa and a lower band with a molecular mass of approximately 135 kDa, indicating that both the core- and the fully glycosylated forms of the protein were phosphorylated.</p><p><b>CONCLUSIONS</b>PKA-mediated phosphorylation of HERG channels causes current reduction in a human cell line. The coupling between the repolarizing cardiac HERG potassium current and the protein kinase A system could contribute to arrhythmogenesis under pathophysiological conditions.</p>

Synthesis and anti alpha-adrenoceptor activity in vitro of 4-((4-((2-substitutedphenoxy)ethyl)-1-piperazinyl) methyl)- 1,2-dihydro-quinoline-2-one compounds / 药学学报

<p><b>AIM</b>To study the synthesis and anti alpha-adrenoceptor activity of 1,2-dihydro-quinoline-2-one compounds.</p><p><b>METHODS</b>Acylation, bromination and cyclization, and substitute reactions were used in the synthesis of the title compounds IV. A proposed mechanism was showed to explain the unusual compounds 5 and 6 in the route C. The inhibition activity of the six target compounds were tested.</p><p><b>RESULTS</b>Twelve new compounds were synthesized (II1-6 and IV1-6). Among them, six new compounds (IV1-6) are the title compounds. Structure of the title compounds were determined by IR, 1HNMR, MS and HRMS.</p><p><b>CONCLUSION</b>Compounds IV3, IV4 and IV6 showed inhibitiion activity, and were worth further studying.</p>

O uso da venlafaxina no tratamento do transtorno do deficit de atencao e hiperatividade / The use of venlafaxine in attention deficit disorder

Uma revisao acerca do uso da venlafaxina no transtorno do deficit de atencao e hiperatividade revela poucos estudos, todos utilizando amostras pequenas. Observou-se eficacia clinica em todos estudos...