Objective To study the effects of Jianpizhidong decoction on dopamine transporter (DAT) mRNA in tourette syndrome(TS) model mice striatum,and provide neural biochemical proof for Jianpizhidong decoction in treating children with TS.Methods 32 male ICR mice were randomly divided into blank control group,model group,Tiapride group,Jianpizhidong decoction group.Except blank control group,the other 3 groups mice were made to TS models by intraperitoneal injection of 3,3'-iminodipropionitrile.The blank and model groups,other 2 groups were given corresponding drugs by intragastric administration respectively.Results The striatum DA TmRNA expression of TS model group,Tiapride group and Tianpizhidong decoction group( respectively(0.17 ± 0.017 ),(0.20 ± 0.021 ),( 0.24 ± 0.022 ) ) decreased than that ( 0.30 ± 0.036 ) of blank control group 13 ％,10％ and 16％ respectively.Conclusion The function of improving symptoms of TS by Jianpizhidongdecoction may be relate to raised DAT gene expression to control the excessive hyperfunction of the DA neurons,and it maybe have better therapeutic than Tiapride.

Objective To study the effects of traditional Chinese medicine of Jianpizhidong decoction on dopamine D2 receptor(DAD2R) mRNA in tourette syndrome (TS) model mice brain striatum,and provide the proof of neural biochemical for Jianpizhidong decoction in curing children TS.Methods 32 male ICR mice were divided randomly into control group,model group,Jianpizhidong decoction group and Tiapride group.Except control group,all mice in model group,Jianpizhidong decoction group and Tiapride group were made to TS models by intraperitoneal injection of 3,3'-iminodipropionitrile.The control gronp and model group were given normal saline,Jianpizhidong decoction group and Tiapride group were given corresponding drugs by intragastric administration respectively.Results The striatum DATmRNA expression of model group,Tiapride group,and traditional Chinese medicine group decreased(respectively(0.139 ± 0.013),(0.15 ± 0.021),(0.141 ± 0.019)) than that of control group (0.180 ± 0.028),and with 14.1％,3.0％ and 3.9％ down respectively.In model group DATmRNA expression was lower than control group (P ＜ 0.01).In model group DATmRNA expression was lower than Tiapride group (P＜ 0.01).In model group DATmRNA expression was lower than traditional Chinese medicine group (P＜0.01).In Tiapride group DATmRNA expression was lower than traditional Chinese medicine group (P ＜ 0.01).Conclusion The function of improving symptoms of TS by Jianpizhidongtang may be relate to inhibite DAD2R hypersensitization to enhance the activity of substantia nigra-striatum.

Objective To study the influences of Jianpizhidong decoction(JPZDD) on bilateral striatal amino acid neurotransmitters in mice with Tourette Syndrome (TS) disease and discuss the mechanism of JPZDD.Methods Thirty mice were injected 3,3-iminodipropionitrile (IDPN) with 350 mg· kg-1 once a day for 7 consecutive days except ten mice of the blank control group which were injected with saline.TS mice were randomly divided into three groups:TS model group,JPZDD group (the Chinese herbal group),Tiapride group (the western medicine group).The TS model group and the blank control group were gavaged with saline(0.9％)at 20 ml ·Kg-1,the Tiapride group with Tiapride at 50 mg · kg-1,and JPZDD group with JPZDD at 20 g · kg-1,respectively,once a day for six weeks.Then behavioral recordings were conducted,and the content of glumatic acid (Glu) and γ-aminobutyric acid (GABA) in the bilateral striatum of the mice were detected by using HPLC.Results Behavioral recording indicated that there were significant differences in the Chinese herbal group and the Tiapride group before(1.90 ± 1.80,2.05 ± 1.40) and after(1.15 ± 1.67,1.00 ± 1.08) treatment (P ＜ 0.05).The content of Glu and GABA in TS model group (1858.33 ± 203.00,261.24 ± 47.31) and Tiapride group (1744.53 ± 234.01,258.57 ± 57.64) were higher compared with the blank control group (P ＜ 0.05),and the content of Glu in JPZDD was significantly higher than that in TS model group.There was no statistical difference between other groups(P＞0.05).The ratio of Glu and GABA were lower in the JPZDD,the Tiapride and the model groups than that in the blank control group,but there were no statistical differences between these groups (P＞ 0.05).Conclusion JPZDD can regulate the content of Glu and GABA in the striatum,and keep the dynamic equilibrium,which might be its pharmacodynamic mechanisms for treating TS.

Objective To study the intervention effect of Jianpi Zhidong Decoction on the expressions of Cx43 and Cx30 in the cortex,corpus striatum,hypothalamus,and hippocampus of tic disorder mice.Methods According to the random number table method,60 male ICR mice were divided into the blank control group(n=15)and modeling group(n=45).The mice in the modeling group were intraperitoneally injected with iminodipropanitrile(350 mg/kg,once a day for 7 days)to establish a tic disorder model,and the mice in the blank control group were intraperitoneally injected with equal volume of normal saline.The stereotyped behavior score was calculated to evaluate the success of modeling.According to the random number table method,the mice in the modeling group were divided into the model group,tiapride group(29.5 mg/kg)and Jianpi Zhidong Decoction group(17.8 g/kg),with 15 mice in each group.The drug was administered by gavage for 4 weeks.After administration,stereotyped behavior score was calculated.The positive protein expression,protein expression,and mRNA expression of Cx43 and Cx30 in the cortex,corpus striatum,hypothalamus and hippocampus were determined by immunohistochemistry,Western blotting and real-time PCR.Results Compared with the blank control group,the stereotyped behavior score of mice in the model group was increased(P＜0.05).Compared with the model group,the stereotyped behavior scores of mice in the tiapride group and the Jianpi Zhidong Decoction group were decreased(P＜0.05).Compared with the blank control group,the average optical densities of Cx43 in the cortex,corpus striatum,hypothalamus,and hippocampus of mice in model group were decreased(P＜0.05).Compared with the model group,the average optical densities of Cx43 in the cortex and hippocampus in the tiapride group were increased,and the average optical densities of Cx43 in the cortex,corpus striatum,and hippocampus in the Jianpi Zhidong Decoction group were increased(P＜0.05).Compared with the tiapride group,the average optical density of Cx43 in the cortex in the Jianpi Zhidong Decoction group was increased(P＜0.05).Compared with the blank control group,the average optical densities of Cx30 in the cortex,corpus striatum,hypothalamus,and hippocampus of mice in model group were decreased(P＜0.05).Compared with the model group,the average optical densities of Cx30 in the corpus striatum and hypothalamus in the tiapride group were increased,and the average optical densities of Cx30 in the cortex,corpus striatum,and hippocampus in the Jianpi Zhidong Decoction group were increased(P＜0.05).Compared with the tiapride group,the average optical density of Cx30 in the hippocampus in the Jianpi Zhidong Decoction group was increased(P＜0.05).Compared with the blank control group,the protein expressions of Cx43 in the cortex,corpus striatum,and hippocampus of mice in the model group were decreased(P＜0.05).Compared with the model group,the protein expression of Cx43 in the cortex in the tiapride group was increased,and the protein expressions of Cx43 in the cortex,corpus striatum,and hippocampus in the Jianpi Zhidong Decoction group were increased(P＜0.05).Compared with the blank control group,the protein expressions of Cx30 in the cortex and hippocampus of mice in the model group were decreased(P＜0.05).Compared with the model group,the protein expressions of Cx30 in the cortex and hippocampus in the tiapride group and the Jianpi Zhidong Decoction group were increased(P＜0.05).Compared with the blank control group,the mRNA expressions of Cx43 in the cortex,corpus striatum and hippocampus in the model group were decreased(P＜0.05).Compared with the model group,the mRNA expressions of Cx43 in the cortex,corpus striatum,and hippocampus in the tiapride group and the Jianpi Zhidong Decoction group were increased(P＜0.05).Compared with the blank control group,the mRNA expressions of Cx30 in the cortex and hippocampus in the model group were decreased(P＜0.01).Compared with the model group,the mRNA expressions of Cx30 in the cortex and hippocampus in the tiapride group and the Jianpi Zhidong Decoction group were increased(P＜0.05).Conclusion The decreased expression of Cx43 and Cx30 in the cortex,corpus striatum,hypothalamus,and hippocampus may be involved in the occurrence of stereotyped behavior in tic disorder mice.The intervention mechanism of Jianpi Zhidong Decoction may be related to the increased expression of Cx43 and Cx30 in various brain regions.

Objective To study the expressions of dynorphin (Dyn) and prodynorphin (PDyn) in cortex and striatum in rats with Tourette syndrome (TS), and interventional effect of Jianpi Zhidong Tang (JZT) on them.Methods Male SD rats (n=60) were randomly divided into model group (n=45) and control group (n=15), and the model group was given intraperitoneal injection of IDPN for establishing TS model.After successful modeling, all rats were taken in the test and randomly divided into model group, Tiapride group and JZT group (each n=15).The model group and control group were given orally normal saline water (0.9%), and other 2 groups were given, respectively, Tiapride (21 mg/kg) and JZT solution (16 g/kg) for 6 w.The integrals of TS rat stereotyped behaviors were recorded after the last drug delivery at every week.The expressions of Dyn and PDyn in cortex and striatum were detected, respectively, by using enzyme-linked immunosorbent assay (ELISA) and real-time fluorescence quantitative polymerase chain reaction (RT-PCR).Results ①The integrals of TS rat stereotyped behaviors showed a descending tend in Tiapride group and JZT group, and the last time observation showed that the integrals were lower in Tiapride group and JZT group than those in model group (P<0.05), and the integrals were lower in JZT group than those in Tiapride group (P<0.05).②The content of cortex Dyn and relative expression of PDyn mRNA had no significant difference among all groups (P>0.05).The content of striatum Dyn and relative expression of PDyn mRNA increased significantly in model group compared with control group (P<0.05), were lower in Tiapride group and JZT group than those in model group (P<0.05), and had no significant difference compared with control group (P>0.05).Conclusion The higher expressions of striatum Dyn and PDyn may take part in TS rat stereotyped behaviors.The anti-tics mechanism of JZT may be related to inhibiting higher expression of striatum Dyn and reducing facilitatory effect in direct pathway.

Objective:To explore the effect of Jian-Pi-Zhi-Dong decoction on autonomous activity and dopamine (dopamine, DA) synaptic vesicle protein expression in the striatum of Tourette syndrome (TS) model rats.Methods:The 4-week-old male SD rats were used to establish the TS model by intraperitoneal injection of N-aminodipropionate. Thirty successfully modeled rats were randomly divided into model group, Chinese medicine group, and tiberide group according to random number table method, with 10 rats in each group. And another 10 rats with matched body mass were selected as the control group. Rats in Chinese medicine group were given Jian-Pi-Zhi-Dong decoction solution (1.6 g/100 g) and the rats in tiapride group were given sulfate tiapride suspension (2.1 mg/100 g), while rats in control group and model group were given an equal volume of 0.9% sodium chloride solution, once a day for 4 weeks.The number of autonomous activities in rats was determined by autonomous activity programmer. ELISA was used to detect the level of DA in the striatum of rats and the expression of dopamine transporter (DAT).Vesicular monoamine transporter-2 (VMAT2) and α-synuclein (α-syn) were measured by Western blot.SPSS 25.0 software was used for data analysis. Multiple group comparisons were performed using one-way ANOVA, non-parametric test and repeated measures ANOVA.Results:Comparing the number of autonomous activities among the 4 groups, the interaction effect between time and group was significant ( F=184.354, P<0.001). At the 1-4 weeks of gavaging, the numbers of autonomic activities in the model group were more than those in the control group (all P<0.05).While the numbers of autonomic activities in Chinese medicine group and tiapride group were less than those in the model group (all P<0.05). Moreover, the numbers of autonomic activities in Chinese medicine group and tiapride group from 1 to 4 weeks were less than those after model making (all P<0.05). The Western blot results showed significant differences in the relative expression of α-syn ( H=29.098), DAT ( F=54.632) and VMAT2 ( H=18.982) among the 4 groups (all P<0.001). The expression levels of α-syn protein in Chinese medicine group and tiapride group were both lower than that in the model group (0.39(0.36, 0.51), 0.39(0.36, 0.50), 0.62(0.50, 0.70)) (both P<0.05). The expression level of DAT protein in Chinese medicine group was higher than that in the model group and lower than that in tiapride group ((0.37±0.06), (0.26±0.07), (0.49±0.09)) (both P<0.05). And the expression level of VMAT2 protein in Chinese medicine group had no significant difference compared with that in the model group ( P>0.05).The ELISA results showed significant differences in DA content of striatum among the 4 groups ( F=75.370, P<0.001). The level of DA in the model group was higher than that in the control group ((7.65±0.72) ng/L, (3.71±0.59) ng/L, P<0.05). The levels of DA in Chinese medicine group ((3.92±0.81) ng/L) and tiapride group ((4.40±0.53) ng/L) were lower than that in the model group (both P<0.05), and the difference between Chinese medicine group and tiapride group was not significant ( P>0.05). Conclusion:Jian-Pi-Zhi-Dong decoction can relieve the tic symptoms of the model rats with TS, and its mechanism may be related to inhibiting the excessive release of α-syn, improving the expression of DAT and VMAT2, improving the DA synaptic vesicle circulation, and reducing the DA content in the synaptic space of the brain.

Objective To establish the quality representation and correlation analysis method and the content of characteristic ingredients based on spectrum of medicinal system of Juemingzi (Semen Cassiae),in order to evaluate the quality of Semen Cassiae decocting pieces effectively and accurately. Methods HPLC-PDA method was established to characterize the quality of spectrum of Semen Cassiae medicinal system architecture;quality of 11 batches of Semen Cassiae decocting pieces was characterized based on the number and chemical type of peaks on characteristic spectrum. The content of 6 characteristic index components, naphthopyrones and anthraquinones were characterized by indexes in characteristic spectrum. Correlation analysis of quality of Semen Cassiae was then performed with reference to the standard reference pieces. Results Taken as the reference substance, characteristic spectrum of batch 10 Semen Cassiae altogether contains 12 characteristic peaks, all characteristic peaks appeared on the chromatograms of 11 batches of Semen Cassiae. In 11 batches of Semen Cassiae, No.5, 11, 8, 1, 2 pieces contained higher effective index components, while No. 5, 11, 2, 9, and 7 pieces had high relevance with the reference piece (No. 10). Consolidated quality characterization and correlation analysis,the quality of No. 5,11,8,1,2,9,and 7 were excellent and dominant. Conclusion The HPLC method to analyze quality and quantity of phenolic components spectrum of 11 batches of Semen Cassiae is simple and accurate. The evaluation mode of quantity and quality and their correalation based on Semen Cassiae medical system, including systemic correlation and appilication validity, as it can provide a reliable methodological basis for the comprehensive quality evaluation of Semen Cassiae.