Objective To improve the sensitivity of MR molecular imaging by using targeting and magnifying effects of biotin-avidin system (BAS). Methods After preparing biotinylated monoclonal antibody HAb18, the number of biotin molecules coupled to each antibody and the binding capacity of biotinylated antibody were determined. Two-step pretargeting tumor imaging was first achieved by intravenously injecting biotinylated monoclonal antibody HAb18 into 8 BALB/c nude mice bearing QGY-7723 tumor cells line. 24 h later, Gd-DTAP-streptavidin (Gd-DTPA-SA) was injected. Gd-DTPA-HAb18 and Gd-DTPA were respectively injected intravenously into the other 12 tumor-bearing mice as contrast groups. MR imaging was performed before and 10, 30, 60 min, 3, 6, 12, 24 h, and 48 h after injection of MR contrast agents. All images were obtained using SE T_1-weighted imaging sequence. After MR imaging, enhancement time course of three different groups was determined by using enhancement data measured in the region of interest in the tumor. Enhancement ratio and contrast-to-noise of tumor were also calculated. Results The average number of biotin conjugated with each monoclonal antibody molecular was 20. And the immunoactivity of biotinylated antibody was 91%. In two-step pretargeting strategy, SI of tumor increased slowly and reached the maximum value at 6 h after injection of Gd-DTPA-SA, enhancement ratio and CNR of tumor had significant difference with other two enhancement methods. The enhancing effect of tumor was still perceptible even after 48 h. When using Gd-DTPA-HAb18, the tumor enhancement pattern was slow and slight. Even at 24 h after injection of Gd-DTPA-HAb18, enhancement ratio of tumor was 13.5%. After Gd-DTPA was injected, signal intensity of tumor increased rapidly, and reached the maximum value at 30 min after injection of Gd-DTPA, and then decreased rapidly. Conclusion Two-step pretargeting strategy based on BAS has specific and signal magnifying effects in tumor MR imaging. It can increase the number of gadolinium that bind to the tumor and provide new approach to MR molecular imaging.

We investigated the current situation about free-ranging dogs infection with plague and provided relevant data for plague prevention and control in Karamay City.We examined the dog serum F1 antibody by indirect hemagglutination assay (IHA).Results showed that the F1 antibody was found in 1 of 13 dogs serum samples,and the positive rate was 7.69％.The free-ranging dogs plays an important role in controlling animal plague epidemic,and there are potential risk for local people's infection of plague.

Objective: To observe the effect of bisoprolol on cardiac function in heart failure (HF) rats and to explore the mechanism. Methods: The experimental rats were divided into 6 groups: Control group, with normal healthy rats, Sham group, the rats received intraperitoneal injection of normal saline; chronic heart failure (CHF) model was successfully established in 40 rats and divided into 4 groups: CHF group, CHF+bisoprolol (Bis) group, CHF+captopril (Cap) group and CHF+Bis and Cap group.n=10 in each group. The cardiac function was observed among different groups; plasma BNP level was measured by ELISA, myocardial miR-25-3p expression was examined by RT-PCR, protein expressions of SERCA2a and phospholamban (PLB) were detected by Western blot analysis and SERCA2a activity was determined by inorganic phosphorus method. Results: Compared with Control group, CHF group showed decreased cardiac output (CO), left ventricular fractional shortening (LVFS), left ventricular ejection fraction (LVEF), reduced expression of cardiac SERCA2a, PLB, the ratio of SERCA2a/PLB and SERCA2a activity; while increased plasma BNP and miR-25-3p expression, allP<0.01. Compared with CHF group, CHF+Bis, CHF+Cap and CHF+Bis and Cap groups had increased CO, LVFS, LVEF, elevated expression of cardiac SERCA2a, PLB, the ratio of SERCA2a/PLB and SERCA2a activity; while decreased plasma BNP and miR-25-3p expression, allP<0.05.Conclusion: Bisoprolol could improve cardiac function in HF rats, which might be related to down regulating myocardial miR-25-3p expression, up regulating myocardial protein expressions of SERCA2a, PLB and enhancing SERCA2a activity.

Objective To investigate the effects of bisoprolol on myocardial SERCA2a activity in rats with heart fail-ure.Methods Male SD rats were randomly divided into normal control group (control group), sham operation group ( sham group ) , model group , bisoprolol group ( Bis group ) , captopril group ( Cap group ) and bisoprolol plus captopril group[(Bis+Cap)group], heart failure rat model was induced by intraperitoneal injections of doxorubicin .Distilled water, bisoprolol, captopril or bisoprolol plus captopril were administrated by gastrogavage for 35 days, respectively. Indices of cardiac function and plasma levels of B-type natriuretic peptide ( BNP) were measured , myocardial expres-sion of miR-25-3p was detected by Stem-loop RT-qPCR, myocardial levels of SERCA2a and phospholamban (PLB) were detected by Western blot , myocardial SERCA2a activity was determined by the inorganic phosphorus method . Results Cardiac function in model group decreased significantly while plasma levels of BNP were significantly higher than those of control group ( P<0.01 ) .Myocardial expression of miR-25-3p in model group was significantly higher while myocardial levels of SERCA 2a and PLB,SERCA2a activity were significantly lower than those of con-trol group(P<0.01).Cardiac function in Bis group , Cap group and Bis +Cap group improved significantly while plasma levels of BNP were significantly lower than those of model group ( P<0.01 ) .Myocardial expression of miR-25-3p in Bis group, Cap group and Bis +Cap group were significantly lower while myocardial levels of SERCA2a and PLB were significantly higher than those in model group (P<0.01).The SERCA2a/PLB ratio and SERCA2a activity in Bis group and Bis +Cap group were significantly higher than those of model group ( P<0.05 ) .Conclu-sions Bisoprolol therapy improves cardiac function in rats with heart failure , which may be related to inhibition of myocardial miR-25-3p, increasing myocardial SERCA2a and PLB levels, enhancing SERCA2a activity.

AIM:To investigate the effects of Xinkang recipe on myocardial miR-25-3p expression and sarco-plasmic reticulum calcium ATPase 2a ( SERCA2a) activity in heart failure rats .METHODS:Male SD rats were randomly divided into normal group , sham group , model group , Xinkang recipe group ( Xinkang group ) , and captopril group .The heart failure rat model was induced by intraperitoneal injection of doxorubicin .Distilled water , Xinkang recipe and capto-pril were administrated by gastric gavage for 35 d, respectively .The indexes of cardiac function and plasma level of brain natriuretic peptide (BNP) were measured.The SERCA2a activity was determined by the inorganic phosphorus method . The myocardial protein expression of SERCA 2a and phospholamban ( PLB) was detected by Western blot .The myocardial expression of miR-25-3p was detected by stem-loop RT-qPCR.RESULTS:Cardiac output (CO), left ventricular fraction-al shortening ( LVFS) and left ventricular ejection fraction ( LVEF) in Xinkang group and captopril group were significantly higher while the plasma levels of BNP were significantly lower than those in model group (P<0.01).The myocardial ex-pression levels of miR-25-3p in Xinkang group and captopril group were significantly lower while the myocardial protein le -vels of SERCA2a and PLB were significantly higher than those in model group (P<0.01).The SERCA2a/PLB ratio and SERCA2a activity in Xinkang group were significantly higher than those in model group (P<0.05), and no significant change was observed between captopril group and model group .CONCLUSION:Xinkang recipe therapy may improve car-diac function in heart failure rats , which may be related to inhibiting the expression of miR-25-3p, increasing the SER-CA2a/PLB ratio and enhancing SERCA 2a activity in the myocardium .

Objective:To construct an adenovirus-mediated anti-sense RNA targeting K-ras exon 1 of SW1990 cell line and observe its effect on ceil proliferation and apoptosis after transferred into SW1990 cell line.Methods:K-ras exon 1 cDNA was cloned into shuttle vector pShuttle-CMV and the resultant plasmid was confirmed by enzyme digestion and PCR.Clones with inverted insertion were selected and co-transferred into E.coli BJ5183 with an adenoviral backbone plasmid pAdEasy-1 to produce recombinant plasmid by homologous recombination.Recombinants were then selected and transfected into 293 cell line to produce recombinant adenovirus.Recombinant adenovirus production was confirmed by PCR analysis and was amplified and purified;the virus titer was determined.Ad-LacZ was used to infect SW1990 cells and the infection efficiency was observed by X-gal staining.SW1990 cells was infected with the recombinant adenovirus and their proliferation and apoptosis were determined by MTT and annexin V/PI FCM assay.Results:A 282 bp target gene fragment was acquired by PCR;the titer of recombinant adenovirus was 7.6?10~8 pfu/ml before purification by CsCl_2 gradient centrifugation and 5.0?10~(10)pfu/ml after CsCl_2 gradient centrifugation.When the recombinant adenovirus was at 100 MOI,the infection efficiency of SW1990 cells nearly reached 100%.The transfection of recombinant adenovirus significantly inhibited SW1990 cell proliferation(P

ABSTRACT:Objective To investigate the effects of Xiefei Lishui recipe on left ventricle remodeling in rats with heart failure.Methods Heart failure rat model was induced by intraperitoneal injections of doxorubicin. Rats were randomly divided into sham operation group (sham group),model group,traditional Chinese medicine group (TCM group),captopril group,and digoxin group.Distilled water,TCM [22 g/(kg · d)],captopril [19 mg/(kg·d)],and digoxin [32μg/(kg·d)]were administered by gastrogavage in rats in different groups for 35 days,respectively.Indices of ventricle remodeling and cardiac function,plasma levels of B-type natriuretic peptide (BNP),rennin (REN),angiotensin Ⅱ(AngⅡ)and aldosterone (ALD)were measured.Cardiomyocyte apoptosis index and collagen volume fraction (CVF)were analyzed.We also assayed myocardial mRNA expressions of MMP-2/9 and TIMP-1/2,and their tissue inhibiting factors TIMP-1 and TIMP-2.Results Compared with those in sham group,in model group cardiac function was significantly decreased,which could be significantly increased by TCM or captopril or digoxin,indices of cardiac remodeling were significantly increased,which could be significantly decreased by TCM or captopril (P<0.01 or P<0.05).Plasma levels of BNP,REN,AngⅡ and ALD,cardiomyocyte apoptosis index and CVF in model group were significantly increased,could be significantly decreased by TCM or captopril (P<0.01 or P<0.05).Myocardial mRNA expressions of MMP-2,MMP-9,TIMP-1 and TIMP-2 in model group were significantly upregulated compared with those in sham group, which could be significantly downregulated by TCM (P<0.01 or P<0.05).Conclusion Xiefei Lishui recipe can attenuate left ventricle remodeling and improve cardiac function in rats with heart failure, which may be related to downregulating myocardial mRNA expressions of MMP-2 ,MMP-9 ,TIMP-1 and TIMP-2 in the left ventricle as well as inhibiting cardiomyocyte apoptosis and myocardial fibrosis.

Objective:To single amino acid mutation of the full human scFvs against TSLP to enhance its affinity.Methods: The specific scFvs against TSLP was screened in our previous study and here the three-dimensional structures of TSLP and anti-TSLP scFvs were simulated by Discovery Studio system,then the molecular docking was made.The amino acids of binding epitope were randomly mutated and the mutated amino acids were selected which could remarkably improve the affinity of scFvs.The primers were designed based on the sequence of mutation amino acids and the scFv sequences were mutated by the overlapping extension PCR.The DNA of mutated scFvs was ligated with the expression vector pLZ16 and transformed into E.coli DH5αF′.Then the scFvs were expressed and the scFvs with improved affinity were selected by ELISA and BIAcore.Results: The five scFvs with single amino acid mutation were screened out by DS system,which could elevate the affinity of scFvs.The mutated anti-TSLP-scFvs were amplified by PCR,which size was about 1 000 bp.The mutated scFvs with correct sequence were expressed,and the mutated scFvs with improved affinity were detected by ELISA and BIAcore.The affinity of selected mutated scFv (M4) has been about 10 times higher than the scFv nonmutation.Conclusion: The affinity of anti-TSLP-scFv has been improved successfully.

Objective To observe the clinical efficacy of plasma exchange(PE),PE combined with plasma bilirubin absorp-tion(PE+PBA),and PE combined with double plasma molecule absorption system(PE+DPMAS),to investigate the best treat-ment options for the patients of hepatitis B-associated subacute severe.Methods Totally 140 patients who had hepatitis B-associat-ed were randomly divided into a PE group,a PE+PBA group,and a PE+DPMAS group.The dinical symptoms and blood rontine, electrolytc and the main biochemical indexes were recorded both before and after treatment and compared among the three groups. Results The total effective rate was higher in the PE +DPMAS group than in the PE group and PE+PBA group,which were 70.8%,60.9%,67.4% respectively,but there was no significant difference(P >0.05).compared with before treatment,serum total bilirubin (TBIL),alanine aminotransferase (ALT),aspartic acid amino shift enzyme (AST),valley aminoacyl transfer peptidase (GGT),alkaline phosphatase (ALP),globulin (GLB)and other biochemical indexes decreased significantly after treatment in 3 groups (P <0.001).PA was increased in the PE group and decreased in the PE+PBA and PE+DPMAS group after treatment,the differences were significant(P <0.05).After treatment,PT was shortened in the PE group,the differences were significant(P <0.05),but there were no significant difference between the PE+PBA and PE+DPMAS groups(P >0.05).Serum K+ and Cl- was obviously declined after treatment in the PE group,there was significant difference(P <0.001).Serum Na+ was obviously declined after treatment between the PE + PBA and PE + DPMAS groups (P < 0.001 ),but the difference was no significant in the PE groups.Serum Ca2+ was significantly decreased in the three groups of patients after treatment(P <0.001).WBC,Hb and PLT were significantly statistical difference after treatment(P <0.05).Conclusion The three groups also can improve liver function and the treatment rate for the patients of subacute hepatitis B-associated severe hepatitis.PE+PBA and PE+DPMAS groups can effective-ly reduce two thirds of the overall usage of the plasma.PE+DPMAS groups that the quantity of Cl- ,Ca2+ ,Hb decline much least in the three groups show larger value in the clinical application.

Objective To study the efficiency of the first time platelet transfusion in patients with blood diseases.Methods Investigate the effect of platelet transfusion in 87 patients with blood diseases.All the patients were transfused 10U platelet (platelet≥2.5×1011 ).Observe CCI after platelet transfusion and analyse the influence factors to the efficiency of the first time platelet transfusion such as the kind of diseases,splenomegaly,fever,age and sex.Results After platelet transfusion,66 patients (75.9 ％) were effective.The efficancy rates were 80.0 ％,78.1％,65.2 ％,82.3 ％ in AA,ITP,AL,MDS (x2=2.88,P＞0.05).In 31 patients with a fever and infaction the efficancy rate was 58.1％,but it was 85.7％ in 56 patients without a fever and infaction (x2=8.3308,P＜0.01).The efficancy rate in 59 patients with non-splenomegaly was 86.4 ％.It was higher than that in 28 patients with splenomegaly which was 53.6％(x2=11.2033,P＜0.01).The efficancy rate was 76.0％ in male patients and it was 75.7 ％ in female patients (x2=- 0.0012,P＞0.05).The efficancy rate was 74.5 ％ in young group (＜60 years old)and 77.8 ％ in old group (≥60 years old)(x2=0.1231,P＞0.05).Conclusion Fever and splenomegaly are the influence factors to the efficiency of the first time platelet transfusion.