Objective To investigate the protective effect of allicin on intestinal mucosal barrier of septic rats so as to explore the possible mechanism.Methods Twenty-four male SD rats were randomly (random number) divided into sham,septic model and allicin treatment group.Septic model was established by cecal ligation and puncture (CLP) in rats.Rats in the treatment group were administered with allicin (30 mg/kg,ip)at 6 h and 12 h after modeling,while those in the model and sham groups were treated with equal amount of saline instead.Rats were sacrificed at 24 h and the serum D-lactic acid,diamine oxidase (DAO) and fluorescence isothiocyanate-dextran (FITC-Dextran,FD-40) were determined to evaluate the intestinal mucosal barrier function.The levels of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),malondialdehyde (MDA),and the activity of superoxide dismutase (SOD) in intestinal tissue were measured.Histopathological changes of intestinal mucosa injury were assessed by Hematoxylin-eosin staining.Results Compared with the sham group,levels of serum D-lactic acid,DAO and FD-40 increased significantly in the CLP group (D-lactic acid:599.4±101.1 vs.149.2±20.63 nmoL/mL,t=11.84,P＜0.01;DAO:302.1 ±64.5 vs.76.57±14.76 ng/mL,t=9.433,P＜0.01;FD-40:6664.0±1437.0vs.1446.0±205.0 ng/mL,t =9.704,P ＜0.01);intestinal morphology damage occurred in the CLP group;intestinal levels of TNF-α,IL-6 and MDA increased greatly (TNF-αt:186.35 ±20.43 vs.58.76 ±8.94 pg/mL,t=17.23,P＜0.01;IL-6:763.25±85.23vs.125.36±14.37 pg/mL,t=22.54,P＜0.01;MDA:29.36±3.27vs.7.24±0.85 nmol/mg prot,t=16.61,P＜0.01),while SOD activity reduced (35.75±6.53 vs.73.26 ±8.35 U/rmg prot,t =10.57,P ＜0.01) in the CLP group.Allicin treatment greatly inhibited the increase of D-lactic acid,DAO and FD-40 levels in rat plasma caused by CLP (D-lactic acid:330.1 ±81.77 vs.599.4±101.1 nmol/mL,t=7.086,P＜0.01;DAO:171.8±49.70vs.302.1±64.56ng/mL,t=5.45,P＜0.01;FD-40:3349.0±1167.0 vs.6664.0±1437.0 ng/mL,t=6.165,P＜0.01);intestinal morphology damage was improved in the allicin treatment group;allicin treatment greatly inhibited the intestinal levels of TNF-o,IL-6 and MDA and preserved the intestinal SOD activity compared with the CLP group (TNF-α:95.37 ±12.68 vs.186.35 ±20.43 pg/mL,t =12.29,P＜0.01;IL-6:354.27±46.27vs.763.25±85.23pg/mL,t=14.45,P＜0.01;MDA:16.27±3.14vs.29.36±3.27 nmol/mgprot,t=9.831,P＜0.01;SOD:55.35 ±6.23vs.35.75±6.53 U/mgprot,t=5.522,P ＜0.01).Conclusions Allicin could inhibit local inflammation and oxidative stress in the intestine and exerts protective effect on intestinal mucosal barrier of septic rats.

Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.

Objective To study the incidence and distribution features of spontaneous decrease of hepatitis B virus (HBV) DNA level and its correlative factors in chronic hepatitis B (CHB) patients.Methods The incidence,demographic features and distribution of different clinical types of spontaneous decrease of HBV DNA were investigated with clinical epidemiological study in 315 CHB or live cirrhosis (LC) patients.The correlative factors of spontaneous decrease of HBV DNA were analyzed by Logistic regression methods.Results Among the 315 patients,171 patients (54.3%) underwent spontaneous decrease of HBV DNA within 12 weeks,of which 61 patients (19.4%) had undeteetable HBV DNA (＜3 lg copy/mL).The stratified study showed that the incidence of spontaneous decrease of HBV DNA in young patients was 58.6% (116/198 cases),which was higher than that (25.0%,2/8 cases) in juvenile patients (X2 = 2.956,P=0.048).The incidence of decrease in relatively more severe patients was higher than that in relatively less severe patients (all X2 in significant groups＞3.84,all P＜0.05),and the highest incidence was 78.7% (48/61 cases) in patients with chronic severe hepatitis B.The incidence of spontaneous decrease of HBV DNA in hepatitis E virus (HEV)/HBV coinfected patients was 75.0% (21/28 cases),which was higher than that (51.8%,146/282 cases) in single HBV-infected patients (X2 =5.530,P=0.019).The incidence of spontaneous decrease of HBV DNA in patients with alanire aminotransferase (ALT)＞400 U/L was 61.8%(102/165 cases),which was higher than that (46.0%,69/150 cases) in patients with ALT≤400 U/L (X2 =7.922,P=0.005).The incidence of spontaneous decrease of HBV DNA in patients with TBil＞200 μmol/L was 68.7% (79/115 cases),which was higher than that (46.0%,92/200 cases) in patients with TBil≤200 μmol/L (X2 = 15.155,P=0.000).The multivariate Logistic analysis demonstrated that chronic severe hepatitis B,ALT＞ 400 U/L and TBil＞ 200 μmol/L were the correlative factors with spontaneous decrease of HBV DNA in CHB or LC patients (all OR＞1,all P＜0.05).Conclusions The spontaneous decrease of HBV DNA level is a common phenomenon during the natural course (immune clearance phase) of CHB or LC patients.Patients with sever conditions,elevated levels of ALT and total bilirubin (TBil) tend to develop spontaneous decrease of HBV DNA.

Objective To observe the protective effects of roscovitine on the podocyte injury induced by endoplasmic reticulum stress (ERS) caused by tunicamycin. Methods The differentiated podocytes cultured at 37℃were randomly di-vided into:(1) Control group, DMSO group and tunicamycin group (TM, 1.0μmol/L). The treatment was given for 3, 6 and 12 hours in three groups. (2) For control group, tunicamycin group, tunicamycin+roscovitine group (20, 40μmol/L, TM+ROS), the treatment was given for 12 hours. The podocyte apoptosis was detected by flow cytometry and TUNEL method. The ex-pressions of Cdk5, GRP78, Caspase-12 and CHOP were detected by Western blot assay. Results (1) Compared with con-trol group and DMSO group, the podocyte apoptosis was increased significantly in a time dependent manner after tunicamy-cin treatment in TM group;the protein expressions of Cdk5, GRP78, Caspase-12 and CHOP were also up-regulated signifi-cantly in TM group (P<0.05). (2) Flow cytometry and TUNEL analysis showed that tunicamycin induced apoptosis in podo-cytes, which was significantly inhibited by roscovitine in a concentration dependent manner in TM+ROS group as compared to that of TM group (P<0.05). The protein expressions of GRP78, Caspase-12 and CHOP were also significantly decreased in a concentration dependent manner in TM+ROS group compared to those of TM group (P<0.05). Conclusion Roscovi-tine, the inhibitor of Cdk5, can reduce the podocyte apoptosis induced by tunicamycin. The protective effects of roscovitine on podocytes can be a novel approach of treating diabetic nephropathy.

Objective To evaluate the effects of hepatitis B virus on liver function,liver fibrosis,and liver pathological staging at different immune stages.Methods We made a retrospective analysis of 657 patients with chronic hepatitis B diagnosed in the First Hospital of Lanzhou University.Their liver function parameters,liver fibrosis parameters,and hepatitis B virus load were measured by automatic biochemical analyzer,automatic gammaradiation immunity analyzer,and quantitative PCR analyzer,respectively.Effects of hepatitis B virus on liver function,liver fibrosis in different immune stages were analyzed by variance analysis.Effects of hepatitis B virus on liver pathological staging at different immune stages were analyzed by linear trend chi square test analysis.Results In ALT normal chronic hepatitis B patients group,viral load had mild effects on liver function and liver fibrosis parameters.However,in ALT abnormal chronic hepatitis B patients group,viral load had a significant effect on liver function and liver fibrosis parameters,and the effect was most obvious in ALT＞double upper limit of normal group.The specific manifestation was that with viral load increasing,liver function parameters including ALT,AST,TBiL,DBiL,and IBiL increased,while TP and ALB decreased.Liver fibrosis parameters HA,LN,PcⅢ,and CIV all increased (P＜0.05).In ALT normal chronic hepatitis B patients group,viral load had no relationship with liver pathological staging.However,in ALT abnormal chronic hepatitis B patients group,especially ALT≥double upper limit of normal group,viral load was significantly related to liver pathological staging.Conclusion The effects of hepatitis B virus on patients' liver function at different immune stages were different,thus providing evidence-based medicine support for clinical antiviral treatment.

Objective:To investigate roles of autophagy in ameliorating sepsis-induced acute lung injury by allicinin in mice.Methods:A total of 152 male Balb/c mice (8-week old) were randomly divided into a sham group,a septic model group,an allicin treatment group,and an autophagy inhibition group.Septic mouse model was established by cecal ligation and puncture (CLP).Mice in the allicin treatment group were given allicin (30 mg/kg,intra-peritoneal injection) at 6 and 12 h,while those in the autophagy inhibition group were given autophagy inhibitor 3-MA (15 mg/kg,intra-peritoneal injection) at half an hour after allicin administration.Mice in the model and sham group were administered with the same amount of saline.Twenty mice in each group were randomly chosen to observe the 7 d survival rate.The other 12 mice were killed at 24 h,and the bronchoalveolar lavage fluid (BALF) (n=6) and lung tissues (n=6) were collected.ELISA was used to detect the tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the BALF.Hematoxylin-eosin staining was preformed to show the morphological changes in the lung tissues.Malondialdehyde (MDA) content and the activity of superoxide dismutase (SOD) in the lung tissues were examined.The expression of LC3B and Beclin-1 was determined by immunohistochemical analysis.Results:Compared with the sham group,the 7 d survival rate and lung SOD activity were decreased in the CLP group (P<0.05);the lung morphological damage score,the levels of TNF-α and IL-6 in the BALF,MDA content in the lung,and expression of LC3B and Beclin-1 were increased greatly in the CLP group (P<0.05).Compared with the CLP group,the 7 d survival rate,lung SOD activity and the expressions of LC3B and Beclin-1 were increased significantly in the allicin treatment group (P<0.05);the lung morphological damage scores,the levels of TNF-α and IL-6 in the BALF and MDA content in the lung were decreased obviously in the allicin treatment group (P<0.05).Compared with the allicin treatment group,the 7 d survival rate,lung SOD activity,and the expressions of LC3B and Beclin-1 were decreased in the 3-MA group (P<0.05);the lung morphological damage scores,the levels of TNF-α and IL-6 in the BALF,and MDA content in the lung were increased significantly in the 3-MA group (P<0.05).Conclusion:Allicin may ameliorate sepsis-induced acute lung injury in mice by enhancing the level of autophagy.

ObjectivesTo identify the related abnormalities of gray matter in pediatric patients with Tourette syndrome (TS) by using the optimized voxel-based morphometry (VBM).Methods Three dimensional T1WI was acquired in 31 TS children (28 boys,3 girts,mean age 8 years,range 4-15 years) and 50 age- and sex-matched controls on a 1.5 Tesla Philips scanner. Images were pre-processed and analyzed using a version of VBM 2 in SPM 2.The whole brain gray matter volume was compared between the study and control group by using t-test.Multivariate linear regression analysis was used for analyzing the correlation between the change of grey matter volume within each brain region (mm3 ) and YGTSS score and course of disease of TS patients.Statistical analyses were performed by using SPSS 13.0.ResultsUsing VBM,significant increases in gray matter volumes in left superior parietal lobule, right cerebellar hemisphere and left parahippocampal gyrus were detected in TS patients,and the volume changes were 4059,2126 and 84 mm3 ( t =3.93,3.71,3.58,P ＜ 0.05 ) respectively.Compared to the control group,decreased grey matter volumes were found in medulla and left pons,and the volume changes were 213 and 117 mm3( t =3.53,3.48,P ＜ 0.05 )respectively.Tic severity was not correlated with any volume changes of gray matter in brain (P ＞ 0.05,a small volume correction,KE ≥ 10 voxel).Tic course was negatively correlated with the gray matter volume of left parahippocampal gyrus ( Beta =- 0.391,P =0.039 ).ConclusionsUsing VBM technique,the gray matter abnormalities can be revealed in TS patients without obvious lesions on conventional MR imaging.The increasing volume of temporal and parietal lobes and cerebellar may be an adaptive anatomical change in response to experiential demand. The gray matter volume of the parahippocampal gyrus may be used as one potential objective index for evaluating the prognosis of TS.

It was estimated that about 428 species of genus Corydalis are distributed all worldwide, with about 298, especially 10 groups and 219 species being uniquely spread in China. The genus Corydalis have been widely employed as folk medicines in China, especially as traditional Tibetan medicines, for treatment of fever, hepatitis, edema, gastritis, cholecystitis, hypertension and other diseases. The phytochemical studies revealed that isoquinoline alkaloids are its major bioactive ingredients. The extensive biological researches suggested its pharmacological activities and clinic applications against cardiovascular diseases and central nervous system, antibacterial activities, analgesic effects, anti-inflammatory, anti-oxidation and anti-injury for hepatocyte, and so on. As an effort in promoting the research of pharmacodynamic ingredients, this article presents an overview focusing on the distribution, phytochemical and pharmacological results of Corydalis species that have been applied in traditional Tibetan medicinal, hopefully to provide a reference for the new Tibetan medicine development from Corydalis plant resource.
Alkaloids ; chemistry ; pharmacology ; Animals ; Anti-Infective Agents ; chemistry ; pharmacology ; Corydalis ; chemistry ; classification ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Mice ; Molecular Structure ; Phytotherapy

Objective:To investigate the related mechanism of salidroside B on proliferation and metastasis of Rhodiola glucoside glioma cell.Methods:The inhibitory rate of salidroside B on SNU-1 cells were determined using MTT method .The effect of salidroside B on inhibiting metastasis of SNU-1 cells was determined by transwell assay .Finally the effect of expression of related proteins of SNU-1 cells was discussed.The SNU-1 cells apoptosis rate was detected by flow cytometry .Results: The growth inhibitory rates of SNU-1 cells were increased with the increasing of dose of salidroside B (10,50,100 μg/ml),and there were significant differ-enced.Apoptosis flow cytometry test results showed that SNU-1 cells apoptosis rate was increased with the increasing of dose of salidroside B(10,50,100 μg/ml).ROS levels was increased with the increase of the concentration of salidroside with significant differ -ence.The expression of salidroside B on Caspase-3,Bax and E-cadherin of SNU-1 cells were increased,Bcl-2,N-cadherin and MMP-9 of SNU-1 cells were decreased.Conclusion:The apoptosis of SNU-1 cells may be through Caspase-3 pathway,mitochondrial pathway apoptosis cascade and the level of ROS by salidroside B , and the reduce of metastasis ability may also through the destruction of adhesion between the cells and basement membrane and increased the stability of tumor cells by salidroside B .

AIMTo prepare the sustained-release nitrendipine microspheres with a solid dispersed structure in liquid system.

METHODSThe sustained-release nitrendipine microspheres with a solid dispersed structure was prepared in liquid system by combining spherical crystallization technique and solvent deposition method in one step. The resultant microspheres were evaluated for the recovery, micromeritc properties, incorporation efficiency. The factors of effect on the formation and the release rate of microspheres were also investigated.

RESULTSThe recovery of microspheres (280-900 microns) was more than 70% and the bulk density was around 0.7 kg.L-1. The incorporation efficiency always exceeded 95%. The formation of microspheres was mainly affected by the amount of bridging liquid and the emulsifying agents in poor solvent. The release rate of nitrendipine from the microspheres could be controlled as desired by adjusting the ratio of talc to Eudragit RS PO in the formulation.

CONCLUSIONThe presented method was suitable for preparing sustained-release microspheres of a water insoluble drug.