Objective To express and purify the N proteins of measles virus(MV)in prokaryotic cells,immunize rabbits to prepare rabbit antisera against MV N proteins,and identify the specificity.Methods Using the genome of MV-S191vaccine strain as template,three fragments of N gene were amplified by PCR to construct the recombinant expression plasmids pGEX-MV-N1,pGEX-MV-N2 and pGEX-MV-N3,which were then transformed into E.coli BL21(DE3)and induced to express the fusion proteins GST-MV-N1,GST-MV-N2 and GST-MV-N3 with GST tags on the N-terminal,respectively. After purification,the fusion proteins were mixed with equal volumes of Freund's complete adjuvant(for initial immunization)and Freund's incomplete adjuvant(for booster immunization),separately,and were used to immunize female rabbits for six times after evenly emulsification,with one rabbit for each protein. The whole blood was collected to isolate the rabbit antisera against MV N proteins,and the specificity of rabbit antisera was identified by Western blot and indirect immunofluorescence assay(IFA).Results The recombinant expression plasmids pGEX-MV-N1,pGEX-MV-N2 and pGEXMV-N3 were constructed correctly as identified by double enzyme digestion,PCR and sequencing. The fusion proteins GSTMV-N1 and GST-MV-N3 mainly existed in the form of inclusion bodies,with a relative molecular mass of about 42 000 and39 000 as well as a purity of 96% and 85%,respectively. However,GST-MV-N2 fusion protein was not detected in the supernatant and precipitate. The rabbit anti-MV-N3 antisera specifically recognized the N protein expressed by MV-S191infected cells,and the specificity was higher than that of rabbit anti-MV-N1 antisera.Conclusion In this study,rabbit antiMV-N antisera was successfully prepared,which provides technical support for the development of recombinant MV vaccine and the preparation of MV structural protein antibody.

Objective To study the clinical characters of coronary artery triple vessel lesion, and to provide a theoretical basis for the therapy to the disease. Methods The clinical data of 68 cases of coronary artery triple vessel lesion (group A) and 71 cases of coronary artery of single or double vessel lesion (group B) were analyzed retrospectively. Results The related risk factors (hypertension, hyperlipidemia, diabetes and smoking) and the clustering of these factors were not significantly different between the two groups (P>0.05). While the positive rate of the exercise stress test existed remarkable difference between the two groups(P<0.01).The extreme differences between the two groups existed also in the cases of building collateral circulation (P<0.001), and in LVEF and LVEDP (P<0.0001) demonstrated by left ventricular angiography. The majority of the coronary artery lesion in group A was on level Ⅳ, and much more lesions of level Ⅲ and level Ⅳ occurred in group A than in group B. Most patients in group A accepted the coronary intervention, while after operation the angina pectoris emerged in a high rate, then the rebuilding of coronary circulation had to be done. Conclusions The best treatment for the patients with coronary artery triple vessel lesion should be the operation of coronary artery bypass graft (CABG), if it's possible.

Objective To study the expression of hypoxia-inducible factor-1 (HIF-1) in the brain tissue after the brain injury caused by acute organophosphate poisoning,and the interventional effect and mechanism of hyperbaric oxygen (HBO) therapy.Methods Sixty healthy male Sprague-Dawley rats randomly divided into a control group (n=6),a poisoning group (n=18),a routine group (n=18) and an HBO group (n=18) according to a random number table.Acute organophosphate poisoning was induced into all rats except those in the control group.The routine group was given penehyclidine hydrochloride and pralidoxime chloride for once,while the HBO group was provided with HBO therapy immediately on the basis of routine treatment.At 1,3 and 7 hours after acute organophosphate poisoning was induced,six rats were sacrificed at each time point and the blood samples were taken from inferior caval vein to measure the content of Malondialdehyde (MDA).The expression of HIF-1α mRNA in the brain tissue was detected by the quantitative real-time PCR,and that of HIF-1 protein was evaluated by immunohistochemical method.Meanwhile,pathologic changes of the brain tissues were observed using hematoxylin-eosin (HE) staining.Results Compared with the poisoning group,the pathological damage to cerebral tissues lessened in the HBO group.The expression of HIF-1 protein and HIF-1 mRNA of the poisoning and the HBO groups was significantly higher than the control group at 3 different time points.After the HBO treatment,the protein expression of HIF-1 lowered from 226.57 ± 57.49,to 205.91 ± 30.36 and further to 187.67 ± 29.25,while the MDA content decreased from 7.74 ± 0.14,to 7.40 ± 0.13 and later to 6.10 ±0.08,both were significantly lower than those of the poisoning group at all time points,with HIF-1 being 1305.67 ± 167.17,2667.83 ± 367.79 and 1709.24 ± 199.07,along with MDA content being 9.48 ± 0.05,11.56 ± 0.13 and 12.26 ± 0.14,and those in the routine group at the time points of 1 and 3 hour later (P ＜ 0.05).A positive correlation was found between the expression level of HIF-1 mRNA and level of MDA in the serum (r =0.909,P=0.000).Conclusion HIF-1 plays an important role in the development of brain injury caused by acute organophosphate poisoning.The efficacy of hyperbaric oxygen intervention against AOPP-induced brain injury is better than that of the routine treatment and its mechanism may be its antioxidation and inhibition of HIF-1 expression.

Objective To observe the changes in the level of serum cystatin C and the expression of matrix metalloproteinase-9 (MMP-9) in the kidney after the development of diabetic nephropathy in streptozotocin (STZ) induced diabetic rats, and to study their correlation. Methods Sixteen female Wistar rats were used to reproduce diabetes by interperitoneal injection of STZ (60mg/kg). Immunohistochemical staining was employed to detect the expressions of MMP-9 in kidneys with image analysis system, and ELISA was used to determine serum cystatin C after 4 or 8 weeks. The correlation between MMP-9 and cystatin C was analyzed by SPSS 10.0. Results The expressions of MMP-9 in diabetic kidneys both at the 4th and the 8th week (8.180?0.905 and 3.948?0.623, respectively) were significantly lower than that of normal control rats (15.620?3.444 and 16.185?3.120, respectively. P

Objective To optimize the extracting process of the mixture of Feining.Methods Orthogonal design was used to optimize extracting process.The content of glycyrrhizicac,Ephedrine and Pseudoephedrin,and the extractive rate of water and n-butyl alcohol were indexes.Results The optimum water-extracting factors were 8 times of water,refluxing and extracting for 1 h,for 2 times.Conclusion The optimal extraction process is precise and repeatable,which can be used in industrial production.

Objective To establish a method for quality control of Jiangya Yishen Granula. Methods Spanishneedles Herb, Radix Polygoni Multiflori, Fruetus Corni, Radix Scrophulariae, Rhizoma Alismatis and Radix Cyathula were identified by TLC. A method of HPLC to determine 2,3,5,4’-tetrahydroxystilbene-2-0-?-D-glucosid, the main effective component from Radix Polygoni Multiflori, was developed. Results The studies on the quality control showed that the characteristics for identification by TLC were distinct. The quantification method had the linear range of 0.016 3～0.522 ?g (r=0.999 9). The average recovery rate was 99.7%, and its RSD was 0.51%. Conclusion The methods can be used for the quality control of the preparation.

Objective To examine plasma levels of MDA-LDL and pregnancy-associated plasma protein-A in patients with angiography type lesions Ⅱ(representing plaque rupture with or without thrombosis), and to evaluate the effect of simvastatin on plasma MDA-LDL and PAPP-A. Methods One hundred and ten patients were enrolled and underwent coronary angiography with 85 patients diagnosed as coronary heart disease (CHD) and twenty-five as controls. According to the morphologic types of plaque, the patients with CHD were straitified as type Ⅰ(smooth borders) (n=31) and type Ⅱ(irregular lesions) (n=35) and type Ⅲ (long lesions with irregular surface) group (n=19). The patients in type Ⅱ group received simvastatin (40 mg/d ) for four weeks. The plasma MDA-LDL, PAPP-A, LDL, HDL levels before and after simvastatin treatment were determined. Results Plasma levels of MDA-LDL, PAPP-A in type Ⅱ group was significantly higer than that in the controls group, type Ⅰ group, type Ⅲ group (P

Purpose To investigate the value of 11C-CFT PET/CT dopamine transporter (DAT) imaging in differential diagnosis of multiple system atrophy (MSA) and Parkinson's disease (PD).Materials and Methods The 11C-CFT PET/CT images of clinically confirmed MSA patients (21 cases),PD patients (24 cases) and healthy adults (10 cases as normal control) were analyzed retrospectively.The volume of interest (VOI) were drawn manually,and the DAT binding indexes and asymmetry indexes of different regions of striatum,including caudate and putamen,were calculated.The differences of DAT binding indexes and asymmetry indexes among the above three groups were analyzed.Results Compared with the normal control group,the striatal DAT binding indexes of MSA group or PD group were significantly reduced (P＜0.05).There were no significant differences in DAT binding indexes between the MSA group and the PD group (P＞0.05).Compared with the normal control group,the DAT binding asymmetry indexes were significantly increased in the PD group (P＜0.05),but the indexes showed no significant differences in MSA group (P＞0.05).The DAT asymmetry indexes of caudate and putamen in the PD group were higher than those in the MSA group (P＜0.05).Conclusion 11C-CFT PET/CT imaging can detect the degeneration of dopaminergic neurons in striatum.The number of striatal dopamine transporters declines in both MSA and PD patients,but the asymmetry of striatal dapamine transporter in PD patients is higher than that in MSA patients.11C-CFT PET/CT can differentiate MSA and PD.

OBJECTIVE:To establish a method for the contents determination of tetrahydropalmatine and rosmarinic acid in Xiaoheling capsule. METHODS:HPLC was performed on the column of Phenomenex C18 with mobile phase A of 0.1% phosphoric acid(adjusted pH to 6.0 with triethylamine)-methanol(45:55,V/V) and (adjusted pH to 6.0 with triethylamine) B of 0.1% phos-phoric acid-methanol(55:45,V/V)(gradient elution)at flow rate of 0.8 ml/min,detection wavelength was 282 nm(0-25 min)and 330 nm(25.01-60 min)and volume size was 10 μl. RESULTS:The linear range was 1.502-75.10 μg/ml for tetrahydropalmatine and 6.03-301.6 μg/ml for rosmarinic acid(r=0.999 8);RSDs of precision,reproducibility and stability tests were ≤1.36%;recoveries were 97.11-99.87%(RSD=0.95%,n=9)and 98.47-101.99%(RSD=1.08%,n=9),respectively. CONCLUSIONS:The method is simple,accurate and reliable,and can be used for contents determination of tetrahydropalmatine and rosmarinic acid in Xiaoheling capsule.

Objective To discuss the therapeutic effect of recombinant human CTLA-4Ig fusion protein (rhCTLA-4Ig) on rheumatoid arthritis (RA) patients and the change of T helper cells 17 (Th1 7 ) and regulatory T (Treg) cells.Methods Forty-eight active RA patients were randomly divided into the treatment group and the control group.Treatment group received 12 weeks of infusions of rhCTLA-4Ig ( 10 mg/kg),while the control group received 12 weeks of infusions of placebo.The changes of expression levels of Th17cell were detected by flow cytometry.The changes of expression level of FoxP3 in monocytes of the peripheral blood were examined by RT-PCR.T test and x2 test were used for statistical analysis.Results ① Eighteen of 24 cases in treatment group showed good ACR20 changes after 12 weeks,the effective rate (75％) was significantly higher than that of the control group ( 1/24,4％ ).After treatment,DAS28 score of the treatment group (3.0±0.7) was significantly lower than that of the control group(6.9±0.7) (t=-12.39,P＜0.01 ).② Compared with the Th17 monocytes of the peripheral blood from active RA patients of treatment group after therapy (0.22±0.20)％,the Th17 monocytes of peripheral blood from active RA patients of the control group (1.63±0.47)％ were higher (t=5.61,P＜0.05 ).③ Compared with the FoxP3 mRNA of the control group (0.24±0.05),that of the treatment group (0.88±0.18) was significantly higher after therapy (t=7.56,P＜0.01 ).Conclusion The clinical parameters of RA patients have improved significantly after the treatment,and the Th17 and Treg cells balance is also restored after rhCTLA-4Ig therapy.