AIM:To explore the effects of endothelial indoleamine 2,3-dioxygenase (IDO) on the migration and the expression of contractile proteins in the pericytes.METHODS: Human pulmonary artery endothelial cells ( HPAECs) and rat cerebral microvascular pericytes were cultured in vitro.Over-expression of IDO in the HPAECs ( IDO-HPAECs) was established.The pericytes were treated with HPAEC-conditioned medium (control group), IDO-HPAEC conditioned medium (treatment group), or IDO-HPAECs-conditioned medium containing 1-methyl-DL-tryptophan (1-mT) ( inhibition group) .The concentrations of nitric oxide ( NO) , tryptophan and kynurenine in the co-culture system were de-termined.The viability, migration and the expression of the contractile proteins in the pericytes were compared.RE-SULTS:No statistical difference of the pericyte viability after treatment with IDO-HPAEC-conditioned medium at 6~48 h was observed (P>0.05).The migratory ability of the pericytes significantly decreased in treatment group compared with control group (P<0.01), and significantly increased in inhibition group compared with treatment group (P<0.01).The concentration of NO in the co-culture system had no significant difference among groups (P>0.05).The concentration of tryptophan was significantly lower in treatment group than that in control group (P<0.01), and significantly higher in in-hibition group than that in treatment group (P<0.01).The concentration of kynurenine was significantly higher in treat-ment group than that in control group (P<0.01), and significantly lower in inhibition group than that in treatment group (P<0.01).The expression ofα-smooth muscle actin and desmin was significantly lower in treatment group than that in control group (P<0.01), and significantly higher in inhibition group than that in treatment group (P<0.01).CON-CLUSION:Endothelial IDO inhibits the migration and the expression of the contractile proteins in the pericytes, and may play essential roles in the regulation of microvasculatures.

Objective To investigate the effect of p15~(INK4B)(p15)gene transfection on the proliferation of pancreatic cancer cell line BxPC3.Methods In p15 transfection group.pCDNA3.1(+)p15 was transfected into BxPC3 cell by the vector of Lipofectamine2000.In empty plasmid transfection group, pCDNA3.1(+)neo was transfected into BxPC3 cell with the sa/ne method as a blank control group.In non-transfection group.the BxPC3 cell was not transfected as a negative control group.The p15 mRNA expressions were assayed by RT-PCR,and p15 protein expressions were assayed by Western blot.The proliferation was determined by MTY assay,ultra-structure changes were measured by transmission electron microscope.Cell cycle and apoptosis were measured by flow cytometry.Results In the pCDNA3.1(+)p15 transfeetion group,the expression of p15 mRNA and protein were resumed.Since the 2nd day of culture,the growth of pCDNA3.1(+)p15 transfeetion group was inhibited,till the 7th day,the inhibitory rate was 47.9%,G_0/G_1 Dhase cell accounted for(61.56±3.96)% of all the cells,which was significantly higher than(47.44±6.35)%ofthe black control groups and(49.22±7.23)%0f tlle negative control group(P<0.05).G_1apoptosis peak occurred and the apoptosis rate was(5.27±1.04)%in pCDNA3.1(+)p15 transfection group,which was significantly higher than(0.11±0.06)% of the black control groups and(0.09±0.07)% of the negative control group(P<0.05).Apoptosis was also observed by transmission electron microscope in the pCDNA3.1(+)-p15 transfection group cells.Conclusions After p15 gene transfection,BLPC3 cell proliferation could be significantly inhibited and apoptosis could be induced.

Objective To produce a comprehensive transcript dataset of Oncomelania hupensis before and after Schistosoma japonicum infection,so as to provide experimental data for perfecting genetic structural information and excavating related mo-lecular markers of O. hupensis infected by S. japonicum. Methods O. hupensis snails were divided into the following 3 groups:one week after S. japonicum miracidium infection,4 weeks after S. japonicum miracidium infection,and normal condition. Mil-lion high-quality reads were obtained from the normalized cDNA of the pooled samples,which were assembled into transcripts. Results A total of 63686 unigenes were identified and were classified into 4 main categories,including general functional pre-diction(15.36%),signal transduction mechanism(11.75%),posttranslational modification(8.89%),and functional unknown (12.20%). Conclusions The transcriptome information of O. hupensis snail after the invasion of S. japonicum shows that sever-al genes are significantly up-regulated or down regulated expression,and that the availability of transcriptome information might provide a strong foundation for further understanding the schistosome-snail interaction at the molecular level.

Objective To figure out the preliminary clinical value of real-time three-dimensional echocardiography(RT-3DE) in evaluating the systolic dyssynchrony in patients with coronary artery heart disease(CAD).Methods Eighty patients who were suspected as CAD were included in this study.RT-3DE was performed first.After the analysis,we got the data:ejection fraction (EF),16 segments systolic dyssynchronic index (SDI 16).All subjects should take the coronary angiography.According to coronary angiography results,the patients with the vascular stenosis rate ≥50% were defined as the CAD group,and the patients with the vascular stenosis rate <50% were defined as the coronary atherosclerosis group,the patients with the completely normal angiographic results were included in the control group according to the results of angiography.The parametric differences among the groups were compared.Results There was no significant difference in LVEF among the three groups.SDI 16 had no significant difference between the control group and the coronary atherosclerosis group (t=-1.03,P>0.05).However,SDI 16 had significant difference between the control group[(3.72±2.68)%]and the CAD group[(7.14±3.10)%],the same between coronary atherosclerosis group[(5.12±3.46)%]and the CAD group[(7.14±3.10)%](t=-3.71,-2.34,all P<0.05).ROC curve analysis revealed that a cut-off value for SDI of 5.49%,yielded a sensitivity of 81.8%,with a specificity of 73.1% to predict coronary atherosclerosis to CAD(AUC=0.743).Conclusion SDI 16 >5.49% has a higher value in evaluating systolic dyssynchrony in patients with CAD with RT-3D of Siemens Acuson SC2000 ultrasonic diagnostic instrument.

Objective To study the preparation and the physicochemical characteristics of glycoprotein from Crassostrea gigas.Methods Oyster collected in Qingdao was taken as the experiment materials,and was extracted with H2O in low temperature.The extracts were purified using Sephacryl s-100 HR gel chromatography and HPLC.Three glycoprotein F22,F33,F42 were finally isolated from the oyster extracts.Results The physicochemical characteristics of glycoprotein were studied.The results showed that the pI value of F22 was 5.5,relative molecular mass of F22 was 34230Da,the protein contents of F22,F33,F42 were 34.1%,19.7% and 12.5%,the saccharide contents of F22,F33,F42 were 35.9%,28.8% and 40%.Conclusion The research result would be used for the study of bioactive component from Crassostrea gigas.

Objective To investigate the effects of trace element strontium on the improvement of rat lipid metabolism disor‐der ,prevention and treatment effects on non‐alcoholic fatty acid liver disease (NAFLD) and its possible mechanism .Methods Fifty SD rats were randomly divided into 5 groups .The control group used the common fodder and the other four groups adopted the high fat fodder for 13‐week feeding .During the final 9 weeks ,the strontium 18 mg/L group and the strontium 36 mg/L group were sepa‐rately fed with 18 mg/L and 36 mg/L of strontium water .During the final 4 weeks ,the simvastatin group was gavaged with simvas‐tatin 10 mg/kg .The rats were killed at the end of 14 weeks and the liver index ,serum ALT ,AST ,TG ,TC ,LDL‐C and HDL‐C ,and liver TG ,TC levels were measured .The liver tissue frozen section was performed .The fatty change and its distribution were ob‐served by oil red O staining .Results Compared with the control group ,the liver indexc ,liver TG and TC levels ,serum TC and LDL‐C in the NAFLD model group were statistically increased (P<0 .05);compared with the NAFLD model group ,the levels of serum TC and LDL‐C in the strontium 18 mg/L group were decreased ,but serum HDL‐C was also decreased(P<0 .05);in liver in‐dex ,liver TC and TG levels ,serum TC and LDL‐C in the strontium 36 mg/L group were decreased(P<0 .05) .The oil red O stai‐ning showed that the liver tissue in the NAFLD model group contained a large amount of red staining fat particles ;but which in the strontium 18 mg/L group ,strontium 36 mg/L group and the simvastatin group were decreased to some extents .Conclusion The long term high concentration trace element strontium intake has the effect for improving the rat lipid metabolic disorder and preven‐ting and treating NAFLD .

Objective To explore the preliminarily clinical value of strain rate parameters using velocity vector imaging (VVI) evaluating left ventricular regional endocardial systolic function in patients with coronary artery heart disease (CAD).Methods A total of eight six inner subjects who were suspected as CAD was enrolled in the study.Patients with the vascular stenosis rate ≥ 50％ were defined as the CAD group,patients with the vascular stenosis rate ＜ 50％ were defined as the coronary atherosclerosis group,and patients with the completely normal angiographic results were included in the control group,according to the results of angiography.The left ventricular endocardial systolic strain rate parameters of VVI were obtained in standard long axis views (apical two,three,and four-chamber view) and short axis views (at the level of the mitral valve,papillary muscles,and apex).The strain rate parameters were global longitudinal endocardial systolic strain rate in the apical two,three,and four-chamber views (A2-GLSRs,A3-GLSRs,and A4-GLSRs),global radial endocardial systolic strain rate in short axis view of the mitral valve level,papillary muscles,and apex (MV-GRSRs,PM-GRSRs,and AP-GRSRs),and global circumferential endocardial systolic strain rate in short axis view of the mitral valve level,papillary muscles,and apex (MV-GCSRs,PM-GCSRs,and AP-GCSRs).The parametric differences were compared among three groups.Results All the subjects included in the present study had normal left ventricular ejection fraction (LVEF) and there was no significant difference in LVEF across three groups.Compared to other groups,the control group had significantly higher E/A ratio.The LV endocardial systolic strain rate parameters were all significantly reduced in the CAD group compared to the control group and the coronary atherosclerosis group (all P ＜ 0.05).Compared to the control group[(-1.37 ± 0.25)/s],the coronary atherosclerosis group [(-1.12 ± 0.42)/s] had significantly lower MV-GCSRs (P ＜0.01).Conclusions VVI is useful for quantitative assessment of the left ventricular systolic function in CAD.MV-GCSRs might have the potential to predict early left ventricle (LV) systolic dysfunction in subjects with coronary artery stenosis ＜ 50％.

Objective To evaluate the usefulness of mycolic acid for identification of Mycobacterium species using SMIS. Methods One hundred and eighteen clinical Mycobacterium isolates collected from Beijing Tuberculosis and Thoracic Tumor Research Institute through whole year of 2007 were analyzed. The 118 isolates contain 25 isolates of Mycobacterium tuberculosis and 93 non tuberculosis Mycobacterium identified by PNB method. Mycolic acid analysis using SMIS is evaluated for identification of a broad range of Mycobacteria in comparison with 16S rDNA , 16-23S rDNA ITS sequencing to measure the concordance rate and agreement, and verify the concordance rate and agreement among results of mycolic acid, sequencing and PNB in identifying Mycobacterium tuberculosis and non tuberculosis Mycobacterium. Results The concordance rate between mycolic acid method analysis and DNA sequencing is 92% ( 108/118), of which concordance rate in Mycobacterium tuberculosis complex and non tuberculosis Mycobacterium are 95% (35/37) and 90% (73/81) respectively, agreement of both is great( agreement Kappa value is 0. 96). Through retrospective analysis, the concordance of results between SMIS and PNB method analysis is 90% (106/118)and agreement is well( agreement Kappa value is 0. 73 ), the concordance of results between sequencing and PNB method analysis is also 90% ( 106/118 ) and agreement is well (agreement Kappa value is 0. 74 ),despite the identification results of 11 isolates by PNB method are discordant. Conclusion Mycolic acid analysis by SMIS enables rapid identification of a broad range of clinical Mycobacterium species, which could play an important role in polyphasic identification of Mycobacterium species.

Objective: To study the gene expression characteristics of steroid- and catecholamine-synthesizing enzymes in adrenal gland of spontaneously hypertensive rats (SHRs), Goto-Kakizaki (GK) rats and normal Wistar rats with the same traditional Chinese medicine syndromes. Methods: Sixteen-week-old Wistar rats, SHRs and GK rats were used. By the quantitative four diagnosis and syndrome differentiation methods and GeneChip Mouse Exon 1.0 ST Array, we observed adrenal gland gene expressions in normal Wistar rats, qi deficiency Wistar rats, SHRs with qi deficiency and qi excess, GK rats with qi deficiency and qi excess. Differentially expressed genes of steroid- and catecholamine-synthesizing enzymes and their regulatory factors were analyzed. Results: Thirty-one genes were differentially expressed among all syndromes. Hsd3b6 was down-regulated significantly 6.0-fold in GK rats with qi deficiency syndrome and qi excess syndrome, and Cyp11b2 was up-regulated 1.5 times in GK rats with qi deficiency syndrome. Por, Hsd11b2, and Nr2f6 were up-regulated in all syndromes, and Cyp2c23, Cyp4a3, Cyp4a8 and Cyp2e1 were down-regulated. However, Srd5a1 and Nr4a1 were up-regulated only in GK rats, and Lss was down-regulated only in SHRs. Th was up-regulated 1.5 times in SHRs with qi deficiency syndrome, GK rats with qi deficiency syndrome and GK rats with qi excess syndrome. Ddc was up-regulated 1.5 times in GK rats with qi excess syndrome. Dbh was up-regulated 3.0 times in GK rats with qi deficiency syndrome and qi excess syndrome. However, Comt was down-regulated 1.5 times in GK rats with qi deficiency syndrome and qi excess syndrome, and Mao was up-regulated 1.5 times in SHRs with qi deficiency syndrome and qi excess syndrome. Conclusion: Some genes associated with steroid- and catecholamine-synthesizing pathways were differentially expressed in SHRs and GK rats, and the differentially expressed genes may be related to the development of traditional Chinese medicine syndromes.

Objective To study the expression features of hydrolase genes related to the secretion of thyroid hormone of H22 hepatoma mice with different symptoms in early stage. Methods Firstly, The quantitative diagnosis and syndrome differentiation methods were used in H22 tumor-bearing mice in early stage, the expression profile of Tg and related hydrolase genes in poisonous pathogenic factors syndrome group (PPFS) and qi-deficiency syndromes (QDS) were got, and the major differential expression were selected. Secondly, the experiment was repeated and ELISA were used to detect T3 and T4 in serum, RT-PCR were applied to detect gene transcription level of genes including Tg, Ctsb, Ctsd, Ctsl, Napsa and Tpp1. Results ① Based on gene chip, the expression of Tg, Ctsb, Ctsd, Ctsl, Napsa and Tpp1were decreased in the first batch of experiment, the exactly ratio was Tg(0.77 in PPFS;0.84 in QDS), Ctsb(0.83 in PPFS, 0.91 in QDS), Ctsd(0.79 in PPFS;no notable change in QDS), Ctsl(no notable change in PPFS; 0.65 in QDS), Napsa(0.78 in PPFS; no notable change in QDS), and Tpp1 (0.75 in PPFS; no notable change in QDS), respectively. ② T3 and T4 downregulated in PPFS (the T3 value was 1.519±0.162ng/ml, T4 value was 2.194±0.305mg/dl) and in QDS (the T4 value is 4.366±0.727μg/dl) in early stage (P<0.01), especially in PPFS, which was in accordance with the change of Tg in both batches. ③the same trend happened in the validation of Tg(0.22 in PPFS;0.38 in QDS), Ctsb(0.31 in PPFS;0.55 in QDS), Ctsd(0.36 in PPFS;0.78 in QDS) and Napsa(0.24 in PPFS;0.59 in QDS) ,while ctsl(1.24 in PPFS;2.11 in QDS) and Tpp1 (2.85 in PPFS;0.85 in QDS)werethe opposite;even this, the total trend of the expression in QDS was still higher than that in PPFS. Conclusion All the results showed that the thyroid function of H22 hepatoma mice was inhibited in early stage especially in PPFS.