A new xanthone from the Polygala tenuifolia Willd. of northern Shaanxi / 药学学报

Ten compounds were isolated and purified from ethanol extracts of dried roots bark of Polygala tenuifolia Willd. by various chromatography techniques such as silica gel and Sephadex LH-20. Their structures were identified by analysis of physicochemical properties and spectral data, and determined as β-sitosterol (1), tenuifolin (2), 6-methoxy coumarin (3), 7-phenyl-1-hydroxy-2,3,6-trimethoxyxanthone (4), 1,8-dihydroxy-3,4,7-trimethoxyanthone (5), mangiferin (6), quercetin-3-O-β-D-glucoside (7), rutin (8), syringaldehyde (9), salicylicacid (10). Among them, compounds 3, 4 and 5 were isolated from the genus of Ploygala for the first time and compound 4 was a new xanthone. The acetylcholinesterase inhibitory activities of compounds 3, 4 and 5 were evaluated by Ellman colorimetric method, compounds 3 and 5 exhibited moderate inhibitory activity, compound 4 exhibited weak inhibitory activity.

Analysis of Inhibitory Effect of Berberine Compounds on Acetylcholinesterase and Blood-brain Barrier Permeability / 中国实验方剂学杂志

ObjectiveTo examine the inhibitory effects of berberine compounds， including columbamine， on acetylcholinesterase from the perspectives of drug-target binding affinity and kinetics and explore the blood-brain barrier （BBB） permeability of these compounds in different multi-component backgrounds. MethodThe median inhibitory concentration （IC50） of acetylcholinesterase by berberine compounds including columbamine was measured using the Ellman-modified spectrophotometric method. The binding kinetic parameters （Koff） of these compounds with acetylcholinesterase were determined using the enzyme activity recovery method. A qualitative analysis of the ability of these components to penetrate the BBB and arrive at the brain tissue in diverse multi-component backgrounds （including medicinal herbs and compound formulas） was conducted using ultra performance liquid chromatography-high resolution mass spectrometry （UPLC-HRMS）. ResultBerberine compounds， including columbamine， exhibited strong inhibition of acetylcholinesterase， with IC50 values in the nanomolar range. Moreover， they displayed better drug-target binding kinetics characteristics （with smaller Koff values） than the positive control of donepezil hydrochloride （P<0.01）， indicating a longer inhibition duration of acetylcholinesterase. Berberine components such as columbamine could penetrate the BBB to arrive at brain tissue in the form of a monomer， as well as in the multi-component backgrounds of Coptis and Phellodendri Chinensis Cortex medicinal extracts and the compound formula Huanglian Jiedutang. ConclusionThese berberine compounds such as columbamine exhibit a strong inhibitory effect on acetylcholinesterase and can arrive at brain tissue in multi-component backgrounds. In the level of pharmacological substance， this supports the clinical efficacy of compound Huanglian Jiedutang in improving Alzheimer's disease， providing data support for elucidating the pharmacological basis of compound Huanglian Jiedutang.

A new pyrazine from Hypecoum erectum L / 药学学报

Four pyrazines were isolated from the n-butanol fraction of Hypecoum erectum L. by using various chromatographic methods, including MCI gel, ODS, silica gel and semi-preparative HPLC. The structures of the isolated compounds were identified as hyperectpyrazin A (1), 1′S-(6-methylpyrazin-2-yl)-ethane-1′,2′-diol (2), 2-hydroxymethyl-6-methylpyrazin (3) and pyrazine-2-carboxylic acid (4) by spectroscopy methods (1D NMR, 2D NMR, UV, IR, MS, etc.). The absolute configuration of compound 2 was determined by using the Mo2(OAc)4 induced CD analysis for the first time. Compound 1 was a new compound, compounds 2-4 were isolated from H. erectum for the first time. Compounds 1-4 were evaluated for their inhibition against acetylcholinesterase and nitric oxide generation induced by lipopolysaccharide-RAW264.7 macrophage cells. At a concentration of 50 μmol·L-1, compounds 2 and 4 displayed inhibitory effects on acetylcholinesterase with the inhibition rates of 44.40% and 43.99%, respectively.

An unprecedented pair of Z/E isomeric pyridinium compound from the aqueous extract of Aspongopus chinensis Dallas / 药学学报

A novel pair of Z/E isomeric compounds with unprecedented carbon skeleton were isolated from an aqueous extract of Aspongopus chinensis Dallas by macroporous resin, silica gel, and semi-preparative high performance liquid chromatography (HPLC). Their structures were identified by nuclear magnetic resonance (NMR), Infrared spectroscopy (IR), Mass spectroscopy (MS) and other spectroscopic methods as (Z)-3-(but-1″-en-1″-yl)-1-(2ʹ-hydroxyethyl)-4-propylpyridin-1-ium, namely aspongopyridine A, and (E)-3-(but-1″-en-1″-yl)-1-(2ʹ-hydroxyethyl)-4-propylpyridin-1-ium, namely aspongopyridine B, respectively. Besides, the anti-inflammatory, anti-tumor, acetylcholinesterase inhibition and butyrylcholinesterase inhibition activities of the compounds 1 and 2 were evaluated. The results showed that compounds 1 and 2 have no anti-inflammatory, anti-tumor, and butyrylcholinesterase inhibition activities instead of weak acetylcholinesterase inhibition activity.

Niveles de colinesterasa sérica en agricultores de San Pablo de Borbur, Boyacá, expuestos a organofosforados

Introducción: Las colinesterasas son enzimas que se encargan de hidrolizar la acetilcolina en ácido acético y colina, poniéndole fin a la transmisión nerviosa a lo largo de la sinapsis de las uniones neuromusculares. La medición de la actividad de la colinesterasa sérica constituye un indicador del efecto causado por la exposición prolongada a los organofosforados. Objetivo: Determinar los niveles de colinesterasa sérica y factores relacionados con la exposición a organofosforados en agricultores de la vereda de Páramo Lagunas de San Pablo de Borbur, Boyacá, Colombia. Metodología: Estudio prospectivo, de corte transversal, con una muestra de 57 trabajadores. A cada individuo se le aplicó una encuesta de datos sociodemográficos y factores laborales, posteriormente se les tomó una muestra de sangre venosa en ayuno de 8-12 horas; con el fin de determinar las concentraciones de colinesterasa sérica (kit Colinesterasa Butiriltiocolina Biosystems®) con el equipo automatizado de química clínica A-15 de Biosystems®. Resultados: El rango de edad de los participantes estuvo entre los 22 y 64 aflos, el 72 % de los individuos pertenecía al sexo masculino. El 3,5 % (2 varones) presentó valores inferiores al intervalo biológico de referencia (IBR), el 88 % de las personas afirmó realizar tareas con plaguicidas y el 54 % afirmó no utilizar los elementos de protección personal (EPP) al trabajar con estas sustancias. Conclusiones: Se evidenció la falta de escolaridad y la ausencia del acompaflamiento técnico en esta zona, lo que induce a que estos agricultores realicen procesos agrícolas relacionados con la aplicación de plaguicidas sin el adecuado conocimiento y sin la utilización adecuada de EPP.

Introduction: Cholinesterases are enzymes responsible for hydrolyzing acetylcholine in acetic acid and choline, which ends nerve transmission along the synapse of neuromuscular junctions. Measurement of serum cholinesterase activity acts as an indicator of the effect caused by prolonged exposure to organophosphates and carbamates Objective: To determine serum cholinesterase levels and factors related to exposure to organophosphates in farmers from the county of Páramo Lagunas in San Pablo de Borbur, Boyacá, Colombia. Methodology: Prospective, cross-sectional study, with a sample of 57 agricultural workers, a survey of sociodemographic data and labor factors was applied to each individual and a venous blood sample was taken in an 8-12 hour fasting, Serum cholinesterase concentrations were determined (Biosystems® Butyrylthiocholine Cholinesterase kit), by means of the Biosystems® A-15 automated clinical chemistry kit. Results: The age range of the participants was between 22 and 64 years old, 72% of the individuals belonged to the male sex. 3.5% (2 male individuals) presented values lower than the biological reference interval (BRI); 88% of the people affirmed to carry out tasks with pesticides and 54% of them affirmed not to use personal protective equipment (PPE) when working with these substances. Conclusions: The lack of schooling was evidenced in most of the farmers, as well as the absence of technical support to this area, which induces these farmers to carry out agricultural processes such as pesticide application, without adequate knowledge and without the proper use of PPE.

Some bis (3-(4-nitrophenyl)acrylamide derivatives: Synthesis, characterization, DFT, antioxidant, antimicrobial properties, molecular docking and molecular dynamics simulation studies-Supplemenntary data

Alzheimer's disease, which is a progressive neurologic disorder, is the most common form of dementia. Although there are various treatment options for Alzheimer’s disease, there is no definite treatment for this disease yet. In this study it was aimed to investigate the treatment potentials of three bis(3-(4-nitrophenyl)acrylamide) derivatives, two of which are known and one is new, for Alzheimer's disease. The study consists of three parts; in the first part of the study, synthesis and characterization studies of the investigated compounds were carried out. In the characterization of the compounds, IR, 1H-NMR, 13C-NMR, LC-MS and elemental analysis techniques were used. In the second part of the study, the compounds were investigated computationally with the assistance of various computational techniques including density functional theory (DFT) calculations, molecular docking and molecular dynamics simulations. In this part, binding free energy calculations were also performed on the investigated compounds. Results of computational studies showed that synthesized compounds interacted with AChE effectively and can be promising structures as AChE inhibitors. In the last part of the study, antioxidant and antimicrobial properties of the compounds were investigated. Antioxidant activities were determined by DPPH? and ABTS?? radical scavenging methods. According to the DPPH? test, the most active compound was found to be 2, while the most active compound was found to be 3 according to the ABTS? test, showing that these methods for antioxidant assay were not significantly correlated with each other. On the other hand, the results of the antimicrobial activity tests showed that compound 3 was the most active compound, which exhibited both antioxidant and antimicrobial activity.

Some bis (3-(4-nitrophenyl)acrylamide derivatives: Synthesis, characterization, DFT, antioxidant, antimicrobial properties, molecular docking and molecular dynamics simulation studies

Alzheimer's disease, which is a progressive neurologic disorder, is the most common form of dementia. Although there are various treatment options for Alzheimer’s disease, there is no definite treatment for this disease yet. In this study it was aimed to investigate the treatment potentials of three bis(3-(4-nitrophenyl)acrylamide) derivatives, two of which are known and one is new, for Alzheimer's disease. The study consists of three parts; in the first part of the study, synthesis and characterization studies of the investigated compounds were carried out. In the characterization of the compounds, IR, 1H-NMR, 13C-NMR, LC-MS and elemental analysis techniques were used. In the second part of the study, the compounds were investigated computationally with the assistance of various computational techniques including density functional theory (DFT) calculations, molecular docking and molecular dynamics simulations. In this part, binding free energy calculations were also performed on the investigated compounds. Results of computational studies showed that synthesized compounds interacted with AChE effectively and can be promising structures as AChE inhibitors. In the last part of the study, antioxidant and antimicrobial properties of the compounds were investigated. Antioxidant activities were determined by DPPH? and ABTS?? radical scavenging methods. According to the DPPH? test, the most active compound was found to be 2, while the most active compound was found to be 3 according to the ABTS? test, showing that these methods for antioxidant assay were not significantly correlated with each other. On the other hand, the results of the antimicrobial activity tests showed that compound 3 was the most active compound, which exhibited both antioxidant and antimicrobial activity.

Endophytic Bacillus amyloliquefaciens surfactin possesses anti-inflammatory potential through acetylcholinesterase and lipoxygenase inhibitory activities

Aims@#This study was aimed to investigate the anti-inflammatory and anti-rheumatoid effects of the Bacillus amyloliquefaciens derived surfactin.@*Methodology and results@#Crude and biosurfactant extracts were analyzed using thin-layer chromatography to determine the presence of biosurfactant. Both extracts were evaluated for their inhibitory effects against the acetylcholinesterase and 5-lipoxygenase enzymes. Human synovial cells were induced with TNF-α and IL-1β. The percentages of the cell viability for both normal and induced cells were determined with an MTT assay. Results showed that surfactin was detected in the biosurfactant extract and demonstrated higher inhibitory effects compared to the crude extract against both inhibitory enzymes acetylcholinesterse (IC50=30.60 μg/mL) and lipoxygenase (IC50=110.10 μg/mL). Both crudes showed no cytotoxic effects at the highest concentration used (50 μg/mL) against normal human synovial cells but showed active reactions against the induced cells. The anti-proliferative effects of biosurfactant and crude extracts were in dose-dependent manner.@*Conclusion, significance and impact of study@#Notably, surfactin obtained from B. amyloliquefaciens has shown an inhibitory effect against pro-inflammatory enzymes and cell viability of the induced rheumatoid arthritis cell line. These results highlighted the therapeutic potential of surfactin application as an anti-inflammatory agent for arthritis treatment. Further study is needed to elucidate the mechanisms underlying the anti-inflammatory effect of surfactin.

Potential of Tetradenia riparia leaf essential oil and its fractions in controlling Aedes aegypti and Rhipicephalus microplus larvae

Tetradenia riparia (Hochst.) Codd (Lamiaceae) is a shrub, commonly known as ginger bush or false myrrh, and several studies have shown that T. riparia exhibits a variety of biological properties. This study aimed to determine the chemical composition of T. riparia essential oil and its fractions, investigate their anticholinesterase activity, and assess their larvicidal activity against the cattle tick Rhipicephalus microplus and the mosquito Aedes aegypti. Eleven essential oil fractions were obtained by fractionation and analyzed by gas chromatography/mass spectrometry. Larvicidal activity against R. microplus and third-instar A. aegypti was assessed using a larval packet test and a larval immersion test, respectively. Anticholinesterase activity was determined by a bioautographic method. Forty-nine compounds were identified in the essential oil, of which the major classes were oxygenated sesquiterpenes (45.95%) and sesquiterpene hydrocarbons (35.20%) and the major components were isospathulenol (17.40%), ß-caryophyllene (15.61%), 14-hydroxy-9-epi-caryophyllene (10.07%), 14-hydroxy-α-muurolene (8.32%), and 9ß,13ß-epoxy-7-abietene (5.53%). Bioassays showed that T. riparia essential oil (LC50 = 1.56 µg/mL) and FR3 (LC50 = 0.30 µg/mL) were the most active against R. microplus and A. aegypti larvae, respectively. The essential oil and FR1, FR2, and FR3 exhibited acetylcholinesterase inhibitory activity. These results indicate that T. riparia essential oil and its fractions hold promise in the development of novel, environmentally safe agents for the control of R. microplus and A. aegypti larvae.

Antioxidant and Acetylcholinesterase Inhibitor Potentials of the Stem Extract of Pternandra galeata / Potenciales antioxidantes e inhibidores de la acetilcolinesterasa del extracto de tallo de Pternandra galeata

Background: Pternandra galeata belongs to the family Melastomataceae. It is a native flowering plant in Borneo Island that serve as food for monkey habitat. There has been limited study on the medicinal and chemical properties of this plant. Objectives: We investigated the acetylcholinesterase inhibitory activity and evaluated the antioxidant activity of the ethanolic extract of Pternandra galeata stem. The total phenolic content in the sample was also determined. Methods: The acetylcholinesterase inhibitory assays were performed using Ellman's method. Two different methods were used to evaluate the antioxidant activity of the extract by 2,2-diphenyl-1-picryl hydrazyl (DPPH) and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. The total phenolic content was determined by the Folin-Ciocalteu method by employing gallic acid as a reference. Results: The ethanolic extract of the P. galeata stems inhibited the AChE enzyme with an IC50 value of 74.62 ± 0.89 µg/mL.The sample exhibited antioxidant activity in the DPPH assay with an IC50 value of 20.21 ± 0.08 µg/mL and 7.68 ± 0.09 µg/mL in the ABTS scavenging assay. The total phenolic content was 164.71 ± 3.33 mg GAE/g extract. Conclusion: The ethanolic extract of the P. galeata stem can be a promising cholinesterase inhibitor and antioxidant for treating Alzheimer's disease

Antecedentes: Pternandra galeata pertenece a la familia Melastomataceae. Se trata de una planta con flores nativa de la isla de Borneo que sirve de alimento para el hábitat de los monos. Se han realizado pocos estudios sobre las propiedades medicinales y químicas de esta planta. Objetivos: Se investigó la actividad inhibidora de la acetilcolinesterasa y se evaluó la actividad antioxidante del extracto etanólico del tallo de Pternandra galeata. También se determinó el contenido fenólico total de la muestra. Métodos: Los ensayos de inhibición de la acetilcolinesterasa (AChE) se realizaron mediante el método de Ellman. Se utilizaron dos métodos diferentes para evaluar la actividad antioxidante de los ensayos de 2,2-difenil-1-picril hidrazilo (DPPH) y 2,2'-azinobis-(ácido 3-etilbenzotiazolina-6-sulfónico) (ABTS). El contenido fenólico total se determinó por el método de Folin-Ciocalteu empleando el ácido gálico como referencia. Resultados: El extracto etanólico de los tallos de P. galeata inhibió la enzima AChE con un valor IC50 de 74.62 ± 0.89 µg/mL. La muestra mostró actividad antioxidante en el ensayo DPPH con un valor IC50 de 20.21 ± 0.08 µg/mL y 7.68 ± 0.09 µg/mL en el ensayo de barrido ABTS. El contenido fenólico total fue de 164.71 ± 3.33 mg GAE/g de extracto. Conclusión: El extracto etanólico del tallo de P. galeata puede ser un prometedor inhibidor de la colinesterasa y antioxidante para el tratamiento de la enfermedad de Alzheimer.

An open-label, randomized study to unfold the potential of Ficus benghalensis in modulation of cognitive impairment using diabetic (Streptozotocin induced) rats

Background: Diabetes mellitus is associated with cognitive, neurophysiological, and structural changes in the central nervous system. Aim and Objective: The aim of the study was to evaluate the effect of Ficus benghalensis on cognitive behavior and acetylcholinesterase levels in brain of diabetic rats, and to compare with Piracetam and Glimepiride. Material and Methods: Wistar rats of either sex weighing 150–200 g were randomized into ten groups of ten each (five groups of diabetic rats and five groups of non-diabetic rats) where one group of diabetic and one group of non-diabetic rats each received F. benghalensis dose I (50 mg/kg), F. benghalensis dose II (100 mg/kg), Piracetam (200 mg/kg) and Glimepiride (0.5 mg/kg), and one group of diabetic rats and one group of non-diabetic rats served as the control group. The blood glucose levels were assessed at 0 and 30th days. The assessment of acquisition phase of each cognitive behavior test was done on 0, 14th, and 29th days, whereas retention phase was assessed on 1st, 15th, and 30th days. Results: In comparison with diabetic control group, F. benghalensis at both doses showed significant decrease in blood glucose levels as well as acquisition and retention of Transfer Latency in elevated plus maze on 29th and 30th days, respectively. Further, both doses exhibited significant increase in retention of step-down latency (SDL) on 30th in continuous avoidance apparatus, but only dose II showed significant increase in acquisition of SDL on 29th day. Similarly, significant increase in retention of Quadrant-time in Morris Water Maze was also observed with both doses of F. benghalensis and other groups compared to controls on 30th day. However, significant decrease in brain AChE level, was observed with only F. benghalensis dose II. Conclusion: Overall, the positive effects of F. benghalensis on cognition were comparable to other two groups, namely, Piracetam and Glimepiride. Hence, it can be concluded that F. benghalensis might be effective in alleviating the behavioral and biochemical changes in diabetes mellitus.

Protective effect of huperzine A on phenytoin induced cognition impairment: Behavioral and biochemical study

Phenytoin, a drug of choice for Epilepsy is also known for its adverse effects. The most common adverse effect due to phenytoin is cognition impairment. Cognition impairment is a serious problem in society as it debars the person's social life. Thus to overcome such a problem demand for a solution arises. Huperzine, sesquiterpene alkaloids having immense neuroprotective properties. Thus in this study, it was aimed to evaluate the effectiveness of huperzine on Phenytoin- induced Cognition Impairment. The protective effect of huperzine on phenytoin-induced cognition impairment was evaluated in rats. The effect of Huperzine on phenytoin-induced cognitive impairment was evaluated by behavioral, biochemical, and histopathological studies. The co-administration of huperzine with phenytoin showed significant results. The treatment of Huperzine with phenytoin resulted in significant improvement in learning and memory. The oxidative stress induced by Phenytoin was reversed by huperzine. A significant decrease in cholinesterase activity was also observed. The histopathology showed damaged neuronal cells in periventricular regions and cortex due to phenytoin which was altered by Huperzine. Thus, the present study demonstrates the protective effect of huperzine on phenytoin-induced cognition impairment.

Effects and mechanism of the ethanol extract of Tiarella polyphylla on learning and memory impairment in mice / 中国药房

OBJECTIV E To investigate the effect s and mechanism of the ethanol extract of Tiarella polyphylla （“TPE”）on learning and memory impairment in mice. METHODS Male Kunming mice were randomly divided into normal group ，model group，positive group （donepezil hydrochloride 4 mg/kg）and TPE low-dose ，medium-dose and high-dose groups （150，300，600 mg/kg），with 10 mice in each group. Drug administration groups were given relevant medicine intragastrically once a day ，and normal group and model group were given water intragastrically once a day ，for consecutive 22 d. On the 17th day ，administration groups and model group were intraperitoneally injected with scopolamine hydrobromide （3 mg/kg）to establish a model of learning and memory impairment. The learning and memory ability of the mice were evaluated by the Morris water maze. Hematoxylin-eosin （HE） staining was used for morphological observation of hippocampus cells of the mice. The levels of acetylcholinesterase （AChE），choline acetyltransferase （ChAT），superoxide dismutase （SOD），malondialdehyde（MDA），interleukin-6（IL-6）and tumor necrosis factor-α（TNF-α）in cerebral tissue as well as the relative expression of phosphorylated Tau protein （p-Tau），β-site amyloid precursor protein cleaving enzyme 1（BACE1）and amyloid precursor protein （APP）in hippocampus tissue were all detected. RESULTS The escape latency of mice in positive group ，TPE medium-dose and high-dose groups were all significantly shortened than the model group on the 4th to 5th day of training ，while the times of crossing platform and the percentage of movement distance in target quadrant were significantly increased （P＜0.05）. Compared with model group ，the neurons in the hippocampal CA 1 region of mice were increased to var ying degrees in administration groups ，the ne urons in solidified and atrophic state decreased ，and the arrangement of neurons tended to be close；the levels of ChAT and SOD in cerebral tissue were significantly increased in positive group and TPE medium-dose and high-dose groups ；the levels of AChE ，MDA，IL-6，the levels of TNF-α and relative expression of p-Tau ，BACE1 and APP in hippocampus tissue were decreased significantly （P＜ 0.05）. CONCLUSIONS TPE can improve the learning and memory impairment induced by scopolamine in mice ，and the mechanism may be related to balancing the brain cholinergic system ，alleviating oxidative stress injury ，improving inflammatory response，and inhibiting the overexpression of p-Tau ，BACE1 and APP .

Anticholinesterase activity of ß-carboline-1, 3, 5-triazine hybrids

Abstract The ß-carboline-1,3,5-triazine hydrochlorides 8-13 were evaluated in vitro against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). The analysed compounds were selective to BuChE, with IC50 values in the range from 1.0-18.8 µM being obtained. The N-{2-[(4,6-dihydrazinyl-1,3,5-triazin-2-yl)amino]ethyl}-1-phenyl-ß-carboline-3-carboxamide (12) was the most potent compound and kinetic studies indicate that it acts as a competitive inhibitor of BuChE. Molecular docking studies show that 12 strongly interacts with the residues of His438 (residue of the catalytic triad) and Trp82 (residue of catalytic anionic site), confirming that this compound competes with the same binding site of the butyrylthiocholine

Vitamin D3 Attenuates Type 3 Diabetic-Associated Cognitive Deficits in Rats through Regulating Neurotrophins and Enhancing Cholinergic Transmission Pathway

Objective: To examine the protective effect of Vitamin D3 against Type 3 diabetes-induced cognitive dysfunction in rats. Materials and Methods: Type 3 diabetes was induced by a high-fat diet plus streptozotocin in rats. Rats were divided into seven groups: negative control, positive control, Vitamin D3 groups (100, 500 and 1000 IU/kg/day), Vitamin D3 plus rivastigmine, and rivastigmine monotherapy. A radial arm maze test was used to assess cognitive function. Levels of acetylcholinesterase (AChE), dopamine (DA), nerve growth factor, neurotrophin-3 (NT-3), and glial cell line-derived neurotrophic factor (GDNF) in the hippocampus were estimated by the enzyme-linked immunosorbent assay kits. Results: Chronic treatment with Vitamin D3 significantly (P < 0.05) and dose dependently alleviated cognitive deficits, with enhancing cholinergic transmission pathway activity through attenuated hippocampal AChE and increased DA level (P < 0.001). Moreover, Vitamin D3 significantly increased (P < 0.001) neurotrophin levels as an underlying mechanism for the resulted improvement. Conclusion: Vitamin D3 plus rivastigmine (combined group) is better than Vitamin D (100 and 500 mg/kg/day) for improvement of AChE, DA, NT-3, and GDNF levels. Vitamin D (500 and 1000 IU/kg/day) was effective as a combined group in terms of the behavioral test.

Kinetic and toxicological effects of synthesized palladium(II) complex on snake venom (Bungarus sindanus) acetylcholinesterase

The venom of the krait (Bungarus sindanus), an Elapidae snake, is highly toxic to humans and contains a great amount of acetylcholinesterase (AChE). The enzyme AChE provokes the hydrolysis of substrate acetylcholine (ACh) in the nervous system and terminates nerve impulse. Different inhibitors inactivate AChE and lead to ACh accumulation and disrupted neurotransmission. Methods: The present study was designed to evaluate the effect of palladium(II) complex as antivenom against krait venom AChE using kinetics methods. Results: Statistical analysis showed that krait venom AChE inhibition decreases with the increase of Pd(II) complex (0.025-0.05 µM) and exerted 61% inhibition against the AChE at a fixed concentration (0.5 mM) of ACh. Kinetic analysis using the Lineweaver Burk plot showed that Pd(II) caused a competitive inhibition. The compound Pd(II) complex binds at the active site of the enzyme. It was observed that K m (Michaelis-Menten constant of AChE-ACh into AChE and product) increased from 0.108 to 0.310 mM (45.74 to 318.35%) and V max remained constant with an increase of Pd(II) complex concentrations. In AChE K Iapp was found to increase from 0.0912 to 0.025 µM (29.82-72.58%) and did not affect the V maxapp with an increase of ACh from (0.05-1 mM). K i (inhibitory constant) was estimated to be 0.029µM for snake venom; while the K m was estimated to be 0.4 mM. The calculated IC50 for Pd(II) complex was found to be 0.043 µM at constant ACh concentration (0.5 mM). Conclusions: The results show that the Pd(II) complex can be deliberated as an inhibitor of AChE.(AU)

Anti-acetylcholinesterase Active Ingredients from Aconitum tanguticum by UPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective:To identify the anti-acetylcholinesterase active ingredients in <italic>Aconitum tanguticum</italic>， so as to lay the foundation for finding new anti-Alzheimer's disease （AD） drugs. Method:The anti-acetylcholinesterase active fractions of <italic>A. tanguticum</italic> were screened by the modified Ellman's method， and the chemical composition of the active fraction was analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS/MS）. The chromatographic separation was performed on an ACQUITY UPLC BEH C₁₈ column （2.1 mm×50 mm， 1.7 μm） with acetonitrile （A）-0.4% ammonia aqueous solution （B） as mobile phase for gradient elution， and the column temperature was set at 30 ℃ with the flow rate of 0.4 mL·min^-1. Phase A of the dichloromethane fraction changed with time as follows：0-3 min， 5%A； 3-7 min， 5%-20%A； 7-11.5 min， 20%-33%A； 11.5-15.5 min， 33%-50%A； 15.5-20.5 min， 50%-80%A； 20.5-23 min， 80%-85%A； 23-25 min， 85%-95%A. Phase A of the <italic>n</italic>-butanol fraction changed with time as follows：0-2 min， 5%A； 2-8 min， 5%-20%A； 8-11 min， 20%-33%A； 11-15 min， 33%-95%A. Mass spectrometry was performed on electrospray ionization， data were collected in positive ion mode， and the detection range was <italic>m</italic>/<italic>z</italic> 100-1 500. Result:Both the dichloromethane and <italic>n</italic>-butanol fractions had a certain inhibitory effect on acetylcholinesterase， their half inhibitory concentration （IC₅₀） values were （64±4.4） mg·L^-1 and （85.7±3.8） mg·L^-1， respectively. By UPLC-Q-TOF-MS/MS analysis， a total of 21 alkaloids were identified from the dichloromethane fraction， and 11 alkaloids were identified from <italic>n</italic>-butanol fraction. Guan-fu base Ⅰ， found in both fractions， was first discovered in <italic>A. tanguticum</italic>. Conclusion:Diterpene alkaloids are the main anti-acetylcholinesterase substances of <italic>A. tanguticum</italic>， which is worth further exploration.

Effect of Forsythionin A on Gastrointestinal Motility Disorders in Mice Treated with Chemotherapy / 中国实验方剂学杂志

Objective:To observe the effect of forsythiaside A on gastrointestinal motility disorder induced by chemotherapy in mice， and explore the mechanism of forsythiaside A regulating gastrointestinal motility. Method:The 60 KM mice were randomly divided into normal group， model group， metoclopramide group （5 mg·kg-1） and forsythiaside A low， medium and high-dose groups （30， 60， 120 mg·kg-1）， 10 for each group， which include half male and half female. The above dose was given once a day for 4 consecutive days， which the intragastric volume was 10 mL·kg-1. One hour after 1rd day administration， equal volume of saline was intraperitoneally injected to the normal group， 2 mg·kg-1 cisplatin was intraperitoneally injected to the other groups with daily for 4 consecutive days. Observing the effects of forsythiaside A on gastric emptying and small intestinal propulsion on mice models， serum gastrin （GAS） and somatostatin （SS）， motilin （MTL）， vasoactive intestinal peptide （VIP） levels were examined by enzyme-linked immunosorbent assay （ELISA）. Activities of acetylcholinesterase （AChE） and total nitric oxide synthase （tNOS） in gastric antrum and ileum were detected by ELISA. The expression of AChE and inducible nitric oxide synthase （iNOS） in gastric antrum and ileum were detected by Western blot. Result:Compared with normal group， the gastric retention rate and small intestinal propulsion rate of the model group were significantly increased （P<0.01）， serum levels of MTL， GAS， SS and VIP， the AChE activity in the homogenate of ileum in the model group were significantly reduced （P<0.05，P<0.01）， while the tNOS activities in gastric antrum and ileum were significantly increased （P<0.05，P<0.01）. Protein expression of AChE in gastric antrum and ileum were significantly decreased （P<0.05）， and the expression level of iNOS protein was significantly increased in the model group （P<0.05）. Compared with model group， different doses of forsythiaside A can reduce the gastric residual rate and small intestinal propulsion rate of mice to varying degrees. Meanwhile forsythiaside A can increase the serum levels of MTL， GAS， SS， and VIP， and the AChE activity and protein expression levels in gastric antrum and ileum tissues were also increased， while tNOS activity and iNOS protein expression were decreased in gastric antrum and ileum （P<0.05，P<0.01）. Conclusion:Forsythiaside A can significantly ameliorate the delayed gastric emptying and small intestine hyperfunction induced by cisplatin in mice. Its mechanism to ameliorate gastrointestinal dysfunction caused by chemotherapy is related to the regulation of gastrointestinal AChE and NOS activity in gastric antrum and ileum and the regulation of gastrointestinal hormone levels.

Diterpenoid alkaloids from roots of Aconitum kongboense / 中国中药杂志

The chemical constituents from the roots of Aconitum kongboense were studied. Twenty-five diterpenoid alkaloids were isolated from the 95% methanol extract of the roots of A. kongboense by silica gel, reverse-phase silica gel and basic alumina column chromatography. They included a new aconitine-type diterpenoid alkaloid, named as kongboensenine(1), and twenty-four known ones(2-25), i.e., acotarine F(2), acotarine G(3), 14-acetyltalatisamine(4), talatisamine(5), indaconitine(6), yunaconitine(7), chasmanine(8), 6-epi-foresticine(9), homochasmanine(10), 8-deacetyl-yunaconitine(11), chasmaconitine(12), ajaconine(13), franchetine(14), ezochasmanine(15), crassicautine(16), 14-O-deacylcrassicausine(17), genicunine A(18), falconeridine(19), sachaconitine(20), liljestrandisine(21), 8-methyl-14-acetyltalatisamine(22), kongboendine(23), 14-benzoylchasmanine(24) and pseudaconine(25). Their structures were elucidated by common spectroscopic methods including high-resolution electrospray ionization mass spectrometry(HR-ESI-MS) and nuclear magnetic resonance(NMR) techniques. Compounds 2-4, 10, 13, 15-19 and 21-22 were isolated from this plant for the first time. Experimental results showed that all compounds did not have a significant inhibitory activity against acetylcholinesterase(AChE).

Expression of / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Acetylcholinesterase (AChE) is a key enzyme used to detect organophosphorus pesticide residues by the enzyme inhibition method. An accidental discovery of a mutant strain with AChE activity was made in our laboratory during the process of AChE expression by