ObjectiveTo explore the efficacy and safety of risperidone and haloperidol in treating Tic disorder.Methods78 patients with Tic disorder were randomly divided into the risperidone group and haloperidol group with 39 cases in each group and treated with risperidone and haloperidol respectively for 8 weeks. All patients of two groups were assessed with the Clinical Global Impression Scale (CGI) and Treatment Emergent Symptom Scale (TESS) before treatment and at the end of the 2nd, 4th and 8th week after treatment. Dosages of patients of two groups were recorded.ResultsAfter 8 weeks treatment, the average maximum dosage of risperidone was (1.4±0.34)mg, and that of haloperidol was (7.3±0.52)mg. The total effective rate of risperidone group was 82% and that of haloperidol group was 82.3 %. There was no significant difference between two groups ( P>0.05). The incidence of adverse reactions in risperidone group was 28.2%, and that in haloperidol group was 76.9%. There was a significant difference between two groups (P<0.01), especially at the end of 2nd week after treatment.ConclusionRisperidone and haloperidol both are effect on Tic disorder, but safety and compliableness of risperidone are higher.

Objective To investigate the effect of histone acetylation change on the transforming growth factor β1 (TGF-β 1)-associated plasminogen activator inhibitor 1 (PAI-1) regulation in mesangial cells (MCs).Methods MCs were transfected with histone acetyltransferase (HAT)CREB-binding protein(CBP),P300 or histone deacetylase (HDAC) 1,3,5 expression vectors,followed by luciferase assay,real-time PCR and Western blotting to see the change of PAI-1 gene's transcriptional activity,mRNA and protein in response to TGF-β1.HDAC inhibitor trichostatin A(TSA)was used and TGF-β1-Smad signaling activity was detected also in this experiment.Results TGF-β1 enhanced PAI-1 transcriptional activity and mRNA expression (P ＜ 0.05).Over-expression of CBP or P300 significantly increased TGF-β1-associated PAI-1 transcriptional activity,mRNA and protein expression (P ＜ 0.05).On the contrary,over-expression of HDAC1,3,5 or dominant negative CBP or dominant negative P300 obviously reduced PAI-1 gene's expression induced by TGF-β1 (P ＜ 0.05).Change of HAT/HDAC did not affect TGF-β1-associated Smad2/3 phosphorylation.TSA enhanced TGF -β1 induced PAI-1 gene's regulation markedly,but did not change TGF-β1-Smad2/3 signaling activity.Conclusion Change of histone acetylation can affect TGF-β1 associated PAI-1 gene's regulation in MCs.

Objective To explore the effect of transforming growth factor β1 (TGF-β1) on epigenetic histone lysine acetylation in the plasminogen activator inhibitor 1 (PAI-1) promoter and transcribe regions in glomerular mesangial cells (GMCs).Methods Chromatin immunoprecipitation assay and real-time quantitative PCR were used to detect Histone3K9 acetylation (H3K9Ac) in the PAI-1 promoter and transcribe regions induced by TGF-β1 and high glucose.Immunoprecipitation was also used to see the cooperation of Smad3,CBP and Sp1 proteins.Results In the four target regions of PAI-1 promoter,TGF-β1 treatment enhanced H3K9Ac at P1,P2 and P3 in GMCs (P ＜ 0.05),but no change was seen in the P4 region which was far from the transcription starting site.TGF-β1 obviously induced H3K9Ac in the T1 transcribe region of PAI-1 instead of T2 (P ＜ 0.05).High glucose increased PAI-1 mRNA expression and H3K9Ac around P1 promoter region (P＜ 0.05).TGF-β1 neutralizing antibody abrogated high glucose-induced H3K9Ac at PAI-1 promoter (P ＜ 0.01).TGF-β1 treatment could recruit Smad3 and CBP protein binding to the PAI-1 promoter regions (P1,P2,P3),and induce their cooperation in GMCs,which were responsing to TGF-β1 associated H3K9Ac.Conclusion TGF-β1 can induce H3K9Ac in the promoter and transcribe regions of PAI-1,promote Smad3 recruition and cooperation with Sp1 and CBP,which are associated with PAI-1 gene's regulation in GMCs.

Objective:To research the best processing method for Bupleurum chinense DC.by orthogonal tests.Methods:With the contents of saikosaponin a and saikosaponin d as the indices,the L9(34) orthogonal table was used to study three factors including the amount of wheat bran,pot temperature before heating and processing time.The orthogonal design was applied to study the processing technology of Bupleurum chinense DC.fried with wheat bran.Results:The best processing method was as follows:100 g Bupleurum chinense DC.was mixed with 10 g wheat bran and fried at 290 ℃ for 80 seconds.Conclusion:The optimized processing technology is reasonable,reliable and highly reproducible,which provide reference for the processing of Bupleurum chinense DC.with wheat bran.

AIM: To test the expression of erythropoietin (Epo) and its receptor EpoR in normal and neovascularized murine corneas induced by alkali burns, and to investigate whether Epo/EpoR is involved in the process of corneal angiogenesis.METHODS: The expression of Epo/EpoR was tested in normal and neovascularized murine corneas induced by alkali burns through immunohistochemistry of corneal frozen sections. Epo cloning, expression, and purification were carried out. Then Epo protein (6μL, 1μg) and control (6μ L of vector control or saline) were injected into the corneal stroma respectively, and the corneas were checked at the 14th day after injection to see whether corneal neovascuarization occurred.RESULTS: Epo/EpoR was expressed in epithelial cells, endothelial cells and stromal cells in normal and neovascularized corneas induced by alkaline burns, and also expressed strongly in neovascularized cornea. They were expressed at the same time in stromal inflammatory cells and new vessels. Corneal neovascularization was induced by Epo intrastromal injection in 5 out of 6 eyes ,but no new vessels were observed in all controls (n = 6) at day 14 after vector control or saline intrastromal injection in normal corneas.CONCLUSION: This paper first reported the expression of Epo and its receptor in normal and neovascularized cornea. Injection of Epo into the corneal stroma may induce the corneal neovascularization. Epo/EpoR is associated with the formation of corneal neovascularization.

Objective:To research the best processing method of ginger juice baking for Rhizoma coptis. Methods:The total con-tent of four alkaloids including berberine hydrochloride determined by HPLC was used as the evaluation index, and an L9(34)orthogo-nal design with three factors including the amount of ginger juice, baking temperature and baking time and variance analysis were ap-plied to study the ginger juice baking technology for Rhizoma coptis. Results:The best processing conditions were as follows:Rhizoma coptis was soaked with 15% ginger juice, baked at 150℃ for 40 min, and withdrawn to be cool. Conclusion:The optimal ginger juice baking technology for Rhizoma coptis is reasonable, which can be used to guide the standardized production of Rhizoma coptis with gin-ger juice baking.

Objective: To observe the clinical efficacy of Tuina (Chinese therapeutic massage) plus needling the distal acupoints along the pathway of meridians for shoulder periarthritis (SP). Methods: A total of 70 patients with SP were divided into an observation group and a control group using the random number table method, with 35 cases in each group. The patients in the observation group were treated by Tuina combined with needling distal acupoints along the pathway of meridians, while those in the control group only received the same Tuina treatment as in the observation group. The visual analog scale (VAS) and shoulder joint function were scored before and after treatment, and the efficacy after treatment was evaluated. Results: After three courses of treatment, the total effective rate of the observation group was 94.3％, while the total effective rate of the control group was 80.0％, showing a statistical difference (P<0.05). After treatment, the VAS scores of the two groups were significantly lower than the baseline scores (P<0.01), and the VAS score of the observation group was lower than that of the control group (P<0.01). After treatment, the total scores of internal rotation, external rotation, back touching, ear touching and motion function in the two groups were significantly higher than those before treatment (P<0.01), and the above five scores in the observation group were significantly higher than those in the control group (P<0.01). Conclusion: Tuina combined with needling the distal acupoints along the pathway of meridians is more effective than Tuina alone in treating SP. The combined therapy can relieve the pain and improve joint function more effectively.

OBJECTIVETo investigate the function of POLH(polymerase eta) through establishment of the POLH gene-blocked cell line FL-POLH(-).

METHODSA mammalian expression vector expressing antisense POLH gene fragment pMAMneo-amp-POLHA (-) was constructed by cloning the 1473 - 2131 fragment of POLH gene into the mammalian expression vector pMAMneo-amp(-) in antisense orientation. The FL cells were transfected with this antisense RNA expressing vector and selected by G418. The mutation assay was conducted using the shuttle plasmid pZ189.

RESULTThe spontaneous mutation frequency of SupF tRNA gene in the plasmid replicated in the FL-POLH(-) was 13.5 x 10(-4), while it was 4.9x10(-4) and 3.7x10(-4) in the control cells FL and FL-M, respectively. The nontargeted mutation frequency of SupF tRNA gene decreased in the plasmid replicated in these cell lines pretreated with MNNG.

CONCLUSIONPOLH plays an important role in maintenance of genetic stability and genesis of nontargeted mutation.

Objective: To investigate the inhibitory effect of Rh2 on HepG2 cells and explore the underlying mechanism.Methods: We used lentivirus carrying β-catenin to infect HepG2 cell, and detected expression of β-catenin using fluorescence microscopy.The effect of Rh2 on proliferation of HepG2-β-catenin and HepG2 cells was measured by CKK-8 assay,and flow cytometry was used to detect cell cycle and apoptosis.The activity of Gsk-3βwas checked by ELISA kit.The expression of Gsk-3β,β-catenin,Bax,Bcl2,CyclinD1,MMP3 genes were measured by qRT-PCR.In order to checked the relationship between β-catenin and TCF4,CHIP assay kit was used,the expression of Bax,Bcl2,CyclinD1,MMP3 genes were measured by PCR.The expressions of Gsk-3β,β-catenin,Bax,Bcl2,CyclinD1,MMP3 proteins were examined by Western blot.Results:HepG2 cells were successfully infected by pLOV-EF1a-MCS-3FLAG-β-catenin lentivirus,named HepG2-β-catenin.CCK-8 showed that ginsenoside Rh2 could effectively inhibit the proliferation of HepG2 and HepG2-β-catenin cells in vitro,which exhibits a dose-dependent manner at range of 10-160 μmol/L Rh2.The IC50 of Rh2 exposure on HepG2 cell for 48,72 h were 100 μmol/L and 58.12 μmol/L,but the IC50 of Rh2 exposure on HepG2-β-catenin for 48,72 h were 129.2 μmol/L,83.33 μmol/L,respectively.The IC50 of Rh2 exposure on HepG2-β-catenin cell was higher than HepG2 cell, compared with HepG2 group the differences was statistically significant ( P<0.01 ).Flow cytometry indicated that Rh2 could arrest HepG2 and HepG2-β-catenin cells in G0/G1 phase;the cell population in G0/G1 phase of HepG2+Rh2 group was(64.57±0.65)%,HepG2-β-catenin+Rh2 group was(58.61±2.01)%.Flow cytometry indicated that Rh2 could induced early apoptosis in HepG2 and HepG2-β-catenin cells.The apoptosis rate of HepG2 +Rh2 group was (17.27 ±2.77)%,HepG2-β-catenin +Rh2 group(9.02 ±1.76)%.The ELISA results indicated that HepG2 cells was induced by Rh2 for 12,24,48,72 h,the activity of Gsk-3βgradually increased,peak in 48 h,then decreased.Compared with control group,Rh2 induced HepG2 and HepG2-β-catenin cells for 48 hours, Gsk-3βactivity were increased, and their activity reduced after adding Bio, there were no significant differences between HepG2+Rh2 and HepG2-β-catenin+Rh2 groups.The PCR,CHIP and WB results showed that the expression of Gsk-3β,Bax gene and proteins increased,while theβ-catenin,CyclinD1,Bcl2,MMP3 gene and proteins down-regulation in HepG2 and HepG2-β-catenin cell induced by Rh2.Compared with HepG2-β-catenin +Rh2 group, the expression of other gene and proteins changed significantly,however,Gsk-3βwas no significant difference.Conclusion:Over-expression of β-catenin may weaken the phar-macological effects of ginsenoside Rh2 on HepG2 cells.The activity of Gsk-3βwas increased by ginsenoside Rh2 to degradeβ-catenin, affecting the expression of downstream genes,promoting apoptosis of liver cancer cells and inhibiting metastasis.

Objective:To study the mechanism of ginsenoside Rh2 inhibiting HepG2 cells migration.Methods:HepG2 cells in logarithmic growth phase were cultured in 96-well plates,which were induced by different concentration Rh2,respectively for 24,48,72 hours.The cell inhibition was detected by Cell Counting Kit.Transwell chambers was used to checked HepG2 cell migration ability;luciferase was tested by Luciferase Reporter Assay system reagent;The expressions of P-ERK,ERK,P-P38,P-38,P-JUK,JUK,MMP3 proteins were detect by Western blot;the expression of AP1,MMP3 gene were detected by Quantitative PCR;The expression of AP1, MMP3 fluorescence protein were observed by fluorescence microscopy.Results:Administrated with different concentration of Rh2 after 24 ,48 ,72 h,the proliferation of HepG2 cells were inhibited ( P<0.05) ,and in dose-and time-dependent manner.Transwell assay showed Rh2 could significantly inhibited migration of HepG2 cells.The expressions of P-ERK , MMP3 proteins were significantly decreased,the expressions of P-JUK, P-P38 proteins were significantly increased, expression levels of ERK, P-38, JUK were no significant difference.Expression of AP1,MMP3 gene were significantly decreased,the expressions of AP1,MMP3 fluorescence proteins were significantly decreased.Conclusion:Ginsenoside Rh2 can activate MAPK pathway to inhibit the migration of HepG2 cells.