Objective: To explore the effects of n-butylphthalide (NBP) on mitochondria in hippocampus and learning and memory abilities in rats with chronic alcoholism. Methods: 60 male SD rats were randomly divided into three groups on average, including normal group, model group and treatment group, with 20 rats in each group.Rats of model group and treatment group are given 6% (V/V) alcohol solution continuously for 28 d to establish the model of chronic alcoholism.Rats in the treatment group were given butylphthalide for 14 days from the fourteenth day after giving alcohol solution.The Y type electric maze was used to test the learning and memory ability of rats, the content of H₂S in the hippocampus and the activity of mitochondrial ATP enzyme were measured by spectrophotometry, and the protein expression of F-actin was detected by Western blot. Results: Compared with the normal group, the learning and memory ability of the rats in the model group were decreased, the content of H₂S in the hippocampus were increased, and the activity of mitochondrial ATP enzyme and the expression of F-actin protein were decreased, and most of the mitochondria were damaged under the electron microscope.The training times of the rats in treatment group(61.88±3.61)was lower than that of the model group(82.19±4.87), the ability of learning and memory was improved(P<0.05). Compared with the model group ((1.50±0.07)U/mgprot, (0.08±0.01)), the activity of the mitochondrial ATP enzyme((1.84±0.11)U/mgprot) and the level of F-actin protein(0.12±0.01)in rat hippocampus of treatment group were increased, the difference was statistically significant(both P<0.05). The level of H₂S in rat hippocampus of the treatment group ((34.56±2.47) nmol/g) was lower than that of the model group ((44.55±3.71) nmol/g), the difference was statistically significant(P<0.05). Compared with model group, the mitochondrial damage of the hippocampus in the treatment group was improved under electron microscope. Conclusion NBP can abate mitochondrial damage and improve learning and memory abilities in chronic alcoholism rats.

The pharmacokinetic profile of gallocatechin-7-gallate (J10688) was studied in rats after intravenous administration. Male and female Sprague-Dawley (SD) rats received 1, 3, and 10 mg/kg (i.v.) of J10688 and plasma drug concentrations were determined by a high performance liquid chromatography-mass spectrometry (LC-MS) method. The pharmacokinetic software Data Analysis System (Version 3.0) was used to calculate the pharmacokinetic parameters. For different i.v. doses of J10688, the mean peak plasma concentration (C 0) values ranged from 11.26 to 50.82 mg/L, and mean area under the concentration-time curve (AUC0-t ) values ranged from 1.75 to 11.80 (mg·h/L). J10688 lacked dose-dependent pharmacokinetic properties within doses between 1 and 10 mg/kg, based on the power model. The method developed in this study was sensitive, precise, and stable. The pharmacokinetic properties of J10688 in SD rats were shown to have rapid distribution and clearance values. These pharmacokinetic results may contribute to an improved understanding of the pharmacological actions of J10688.

Objective To observe the changes of N-methyl-D-aspartate (NMDA) receptor 2B subunit (NR2B) expression in the striatum of chronic alcohol exposured rats at different withdrawal time.Methods 72 male Wistar rats were randomly divided into withdrawal 2h group,withdrawal 6h group,withdrawal 12h group,withdrawal 1d group,withdrawal 3d group and control group,and 12 rats in each group.In the 5 withdrawal groups,ethanol was administered in drinking water at the concentration of 6％ (V/V) for 16 weeks,and rats in control group were maintained with water.After 16 weeks ethanol was removed and ethanol withdrawal syndromes were evaluated.The expression of NR2B protein in the striatum was measured by immunofluorescence and western blot and the expression of NR2B mRNA in the striatum was measured by realtime PCR.Results Compared with withdrawal scores of control group((1.50±0.80)),scores of withdrawal 2h,6h,12h,1d,3d groups ((10.42±2.50),(15.42± 1.93),(9.25±2.01),(7.67± 1.92),(2.25±0.87) respectively) were higher,and the withdrawal scores of withdrawal 6h group were the highest.Compared with the expression of NR2B fluorescence intensity (2210.00± 178.20),the expression of NR2B protein(0.150±0.009) and the expression of NR2B mRNA(0.006±0.001) in the striatum of control group,the expression of NR2B fluorescence intensity (2710.56 ± 194.21),(5035.16 ± 234.41),(3326.23 ± 378.16),(2570.64 ±177.88),the expression of NR2B protein (0.192±0.008),(1.649±0.205),(0.783±0.109),(0.180±0.009) and the expression of NR2B mRNA (0.026±0.002),(0.351±0.034),(0.248± 0.023),(0.024±0.003) of withdrawal 2h,6h,12h,ld groups were significantly higher (P＜0.05),and with the extension of the withdrawal time,the expression was gradually increased.The expression of withdrawal 6h group was the highest,then began to decline,and returned to baseline levels at withdrawal 3 d(P＞0.05).Withdrawal scores were positively correlated with the expression of NR2B protein(r=0.719,P＜0.01),the expression of NR2B protein was positively correlated with the expression of NR2B mRNA(r=0.937,P＜0.01),and the expression of NR2B mRNA was positively correlated with withdrawal scores(r=0.673,P＜0.01).Conclusion The expression of NR2B was up-regulated in the striatum of chronic alcohol exposured rats at different withdrawl time.NR2B protein and NR2B mRNA expression is positively correlated with the withdrawal scores,suggesting that regulating the expression of NR2B may be a new target for the treatment of ethanol withdrawal symptoms.

In order to improve the efficiency of drug screening on serotonin transporter (SERT) inhibitors, a high-throughput screening (HTS) model is established in RBL-2H3 cells. The RBL-2H3 cells are very similar to the serotonin genetic neuro, in modulation of post-receptor mechanisms and transduction pathway of SERT reactivated. Depending on a fluorescence substrate ASP+ used in detection method of inhibitor rates, it's convenient, quick, accurate and effective, not making the environmental biohazard compared with radioactive experiments. Furthermore, biological screening model combined with computer aided virtual screening technique describing high-throughput virtual screening (HTVS). Bayesian classification method and molecular fingerprint similarity were applied to virtual screening technique, for screening compounds in compound library. Some compounds have been found, and then validated further by biological screening model. Combination of HTS and HTVS improves the efficiency of screening SERT inhibitors.

The emerging of network pharmacology and polypharmacology forces the scientists to recognize and explore new mechanisms of existing drugs. The drug target prediction can play a key significance on the elucidation of the molecular mechanism of drugs and drug reposition. In this paper, we systematically review the existing approaches to the prediction of biological targets of small molecule based on chemoinformatics, including ligand-based prediction, receptor-based prediction and data mining-based prediction. We also depict the strength of these methods as well as their applications, and put forward their developing direction.

Objective To observe the effect of quetiapine on levels of glutamate and expression of cannabinoid receptor type 1 (CB1) in the hippocampus of rats with chronic alcoholism.Methods Fortyeight rats were randomly divided into 4 groups of 12:control group,chronic alcoholism group,low and high quetiapine groups (both with chronic alcoholism),to establish an animal model of chronic alcoholism.The abstinence scoring was used to evaluate the rats' withdrawal symptom; the concentrations of glutamate in rats' hippocampus were measured by high-performance liquid chromatography (HPLC) ; expression of CB1 mRNA was evaluated by real-time quantitative PCR,CB1 protein expression in the hippocampus was detected by immunofluorescence.Results The abstinence score (11.20 ± 3.01 vs.5.50 ± 1.90),glutamate content ((0.26 ± 0.06) μmol/g vs.(3.55 ± 0.67) μmol/g),CB1 mRNA relative copy numbers (7.53 ± 0.80 vs.1.83 ± 0.81) were significantly different in the chronic alcoholism group as compared to the control group (t =5.06,15.52,15.88,all P ＜ 0.05).The abstinence score (6.70 ± 1.34 vs.11.20 ± 3.01),glutamate content((2.56 ± 0.55) μmol/g vs.(0.26 ±0.06) μmoL/g),CB1 mRNA relative copy numbers (1.95 ± 0.65 vs.7.53 ± 0.80) were also significantly different in the high quetiapine group as compared to the chronic alcoholism group (t =4.32,13.19,17.11,all P ＜ 0.05) ; immunofluorescence showed that CB1 positive cells in the chronic alcoholism group were significantly increased compared with the control group,and CB1 positive cells in the high quetiapine group were significantly decreased compared with the chronic alcoholism group.Conclusion Quetiapine may alleviate the damage from chronic alcoholism,the mechanism may be relate to the regulation of the CB1/glutamate pathway.

Objective To observe the effect of quetiapine on levels of glutamate and expression of cannabinoid receptor type 1 (CB1) in the hippocampus of rats with chronic alcoholism.Methods Fortyeight rats were randomly divided into 4 groups of 12:control group,chronic alcoholism group,low and high quetiapine groups (both with chronic alcoholism),to establish an animal model of chronic alcoholism.The abstinence scoring was used to evaluate the rats' withdrawal symptom; the concentrations of glutamate in rats' hippocampus were measured by high-performance liquid chromatography (HPLC) ; expression of CB1 mRNA was evaluated by real-time quantitative PCR,CB1 protein expression in the hippocampus was detected by immunofluorescence.Results The abstinence score (11.20 ± 3.01 vs.5.50 ± 1.90),glutamate content ((0.26 ± 0.06) μmol/g vs.(3.55 ± 0.67) μmol/g),CB1 mRNA relative copy numbers (7.53 ± 0.80 vs.1.83 ± 0.81) were significantly different in the chronic alcoholism group as compared to the control group (t =5.06,15.52,15.88,all P ＜ 0.05).The abstinence score (6.70 ± 1.34 vs.11.20 ± 3.01),glutamate content((2.56 ± 0.55) μmol/g vs.(0.26 ±0.06) μmoL/g),CB1 mRNA relative copy numbers (1.95 ± 0.65 vs.7.53 ± 0.80) were also significantly different in the high quetiapine group as compared to the chronic alcoholism group (t =4.32,13.19,17.11,all P ＜ 0.05) ; immunofluorescence showed that CB1 positive cells in the chronic alcoholism group were significantly increased compared with the control group,and CB1 positive cells in the high quetiapine group were significantly decreased compared with the chronic alcoholism group.Conclusion Quetiapine may alleviate the damage from chronic alcoholism,the mechanism may be relate to the regulation of the CB1/glutamate pathway.

ObjectiveTo observe the effect of alcohol exposure during pregnancy on learning and memory and content of hydrogen sulfide(H2S) in the hippocampus of infant rats.MethodsThe animal models of alcohol exposure during pregnancy were made,and the learning and memory were evaluated by Y-maze in adult offspring.Content of H2S and activity of cystathionine-beta-synthase(GBS) in the hippocampus of the brain were evaluated with spectrophotometry;and CBS protein expression in the hippocampus was detected by immunohistochemistry.ResultsThe learning and memory ( (43.00 ± 15.33 ) times) of alcohol exposure during pregnancy group was significantly decreased compared with that of control and drinking groups (( 25.13 ± 12.35 )times and (26.12 ±11.95 ) times,P ＜ 0.05 ) ; spectrophotometry results showed that the content of H2S ( ( 30.32 ± 5.84 ) nmoL/g) of alcohol exposure during pregnancy group was significantly increased compared with that of control ( ( 52.51 ±7.85 ) nmol/g) and drinking groups( (49.93 ± 4.29 ) nmol/g),and the activity of CBS( ( 55.13 ± 4.45 ) nmol/g)of alcohol exposure during pregnancy group was significantly increased (P ＜ 0.01 ) compared with that of control ( (71.06 ± 5.58 ) nmol/g) and drinking groups( (69.96 ± 6.13 ) nmol/g) ; immunohistochemistry showed that the expressionof CBSproteinofalcoholexposureduringpregnancygroupwassignificantlyincreased.ConclusionThe damage effect of alcohol exposure during pregnancy on nerve system of infant rats may interrelate with down-regulation of H2S/CBS in the hippocampus.

In present study, standard method and standard operation practice for measuring the activities of influenza neuraminidase and its inhibitors have been established. The accuracy and stability of the method has been evaluated. Standard operation is as following: 10 microL sample, 30 microL neuraminidase and 60 microL substrate are added to one well of a 96-well plate, and then incubated at 37 degrees C for 1 h. The reaction was stopped with NaOH before fluorescence intensity determination. One unit of neuraminidase is defined as the amount of enzyme that produces 1 nmol 4-MU in 1 h under above conditions. The inhibition accuracy is indicated by an uncertainty measurement of 6.51 x 10(-12), and its stability was reaffirmed by determination of oseltamivir acid. In this study, systematic assessment of neuraminidase inhibitory assay not only provided theoretical basis of its application in drug discovery, but also made preliminary attempt to use uncertainty measurement as a parameter in biological measurement.

Objective To investigate the effect of butylphthalide (NBP) on H2S content and the expression of NR2B in the hippocampus of alcohol dependence rats. Methods A total of 84 SD male rats were randomly divided into 6 groups. Except for the normal group, other groups were subjected to alcohol solution with concentration of 6% ( V/V) for 28 d. Drug intervention began at the 14th day,and rats in the low,medium,high dose group were treated with NBP with a different concentration. Erden abstinence scoring was used to evaluate the rats withdrawal symptom. H2S content was measured in one side of hippocampus and CBS activity was tested in the other side of hippocampus. Hippocampus of 3 rats from each group was used to investigate NR2B mRNA level. Results Withdrawal symptom score ( 12.27 ± 1. 19),H2S content(30. 25 ±8.82), CBS activity (72. 44 ±7. 46) and NR2B mRNA expression( 19. 47 ±0. 86) in medium dose NBP group rats were lower than withdrawal symptom score(14.09 ±2.21) ,H2S content(44. 50 ±6. 65) , CBS activity(79. 06 ±4. 57) and NR2B mRNA expression (29. 13 ±1.39) in experimental control group (P<0.05). Withdrawal symptom score(12. 18 ±1.08) ,H2S content(33.00 ±5.38) ,CBS activity(67. 81 ±9. 37) and NR2B mRNA expression(23. 12 ± 1. 86) in high dose NBP group rats were lower than experimental control group (P < 0. 05). Conclusion NBP can reduce withdrawal symptoms of alcohol dependence rats,may be related to decreased expression of H2S/CBS system, and NR2B mRNA expression.