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Contrast induced nephropathy (CIN)is one of the main complications caused by intravenous iodinated contrast media injection,which can increase hospitalization rate and mortality of patients,and it has become the third main cause of acquired acute renal failure in hospital currently, but its fundamental pathogenesis is still not clear,which is considered to be related to inflammation and oxidative stress response etc.The present article made a review on pathogenesis of CIN and preventive effects of statins on CIN.
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Objective:To explore whether cigarette smoking extract (CSE) has influence on fluorescence protein ex-pression of thrombomodulin (TM) on surface of African green monkey kidney fibroblast cells (COS-7) or not . Methods:When TM-green fluorescent protein (GFP) plasmid was successfully constructed ,COS-7 cells were trans-fected by it ,then incubated by prepared 5% CSE (5% CSE group) ,PBS of certain volume was added to serum-free minimal essential medium (MEM ,simple medium) ,which was cultured at the same time and treated as control group .Flow cytometry counting method was used to detect change of TM-GFP expression amount on COS-7 surface at different time point .Results:Compare with control group ,there were no significant difference in the expression of TM-GFP on COS-7 surface at 1h and 6h in 5% CSE group [1h :(134.99 ± 18.41) vs .(146.61 ± 12.06) ,6h :(116.89 ± 27.28) vs .(123.89 ± 39.24) ,P>0.05 both] .Conclusion:The 5% cigarette smoking extract has no influ-ence on fluorescence expression of thrombomodulin on surface of African green monkey kidney fibroblast cells .
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Aim To study the effect of CSE ( cigarette smoke extract ) on the single-molecule interactional force between thrombomodulin and thrombin by live-cell single-molecule force spectroscopy. Methods CSE was prepared by a previously reported method. The plasmid of TM-GFP was constructed and transfect-ed in COS-7 cells. The expression of TM-GFP was de-tected by fluorescence microscopy and laser scanning confocal microscopy. The transfected COS-7 cells were grouped ( 1 ) GFP -thrombin group ( 2 ) TM-thrombin group ( 3 ) CSE-TM-thrombin group ( 4 ) CSE- GFP-thrombin group. Force measurements with the thrombin modified AFM tips on the living cell surface were car-ried out on PicoSPM II with a Pico-Scan 3000 control-ler and a larger scanner. The force curves measured in living cells were recorded by PicoScan 5 software and analyzed by MATLAB R2009aMetlab. Results The single-molecule binding force of thrombomodulin and thrombin ( TM-Thr ) was determined ( 60. 90 ± 0. 82 ) pN. The binding probability for TM-Thr was about (22. 58 ± 3. 95)%. Antibody blocking binding proba-bility for TM-Thr was ( 2. 58 ± 2. 0 )%. The binding probabilities for GFP-Thr group, CSE-TM-Thr group and CSE-GFP-Thr group were significantly decreased compared with TM-Thr group ( P<0. 05 ) . The mean value of the most probable single molecular interaction force of thrombin/TM-ECD was determined as ( 45. 30 ± 1. 37 ) pN, the binding probability of thrombin and TM-ECD was ( 23. 25 ± 7. 02 )%. When the binding was blocked with the TM-MAb solution, the binding probability decreased to ( 4. 64 ± 2. 31 )%. The bind-ing probability was ( 8. 31 ± 1. 06 )% in the CSE-TM-thr-S group. When further blocked with TM-MAb, the binding probability was ( 5. 17 ± 2. 96 )%. Conclusion CSE significantly decreases the binding probability for TM-Thr to induce intravascular thrombosis.
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The gene expressions of codeinone reductase (COR) and berberine bridge enzyme (BBE) in Papaver somniferum were blocked by RNA hairpin of RNA interference (RNAi). The complete sequences of COR and BBE genes were cloned by reverse transcription-polymerase chain reaction (RT-PCR), the results of homology comparison revealed that the cloned COR and BBE genes had high homology with the other gene family members reported in the GenBank. The target sequences of COR and BBE genes were screened in accordance with the design principle of RNAi, a 643 bp fusion gene was obtained by the method of overlapping PCR, then plant expression vector ihpRNA was constructed based on intermediate vector pHANNIBAL and plant expression vector pCEPSPS. With that 78 transgenic plants were obtained through Agrobacterium-mediated and 17 positive plants were screened by PCR, that could initially indicate that the target fragments of COR and BBE gene had been integrated into tobacco genome.
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The codeinone reductase gene (cor) and the berberine bridge enzyme gene (bbe) were cloned from young leaf of opium poppy(Papaver somniferum L) by RT-PCR and the coding sequences of these gene were analyzed. The result demonstrated that the cloned COR gene sequence was highly homologous to the other COR gene family members showing 98.96% identity to COR1.1. The cloned BBE gene sequence was 94.84% identified with the released BBE genes in GenBank previously. Based on the cDNA sequences of COR and BBE,two fragments about 400～500 bp from each gene with lower identity among them were cloned. The fusion gene BC(744 bp) is fused by the PCR technique. Then the promoter CaMV 35S driven,containing'forward BC fusion fragments-reverse pdk intron-reverse BC fusion fragments',plant siRNA expression vector were constructed based on the vectors pHANNIBAL and pART27. Inhibition efficiency of the expression vector to the morphine synthesis was studied by transforming papaver nudicarule.The work will lay the foundation for breeding a low morphine and high thebaine poppy.
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Objective To investingate the role of inflammation in coronary atherosclerosis and the relationship between the inflammatory factors and coronary risk score. Methods 56 patients with acute coronary syndrome were studied, among them 26 cases were diagnosed as acute myocardial infarction (AMI) and 30 unstable angina pectoris (UAP). The study group was compared with a control group of 30 cases who were identified as normal by coronary angiography. The concentrations of serum sICAM-1 and hsCRP were determined by ELISA assay. The coronary risk score was recorded in patients with UAP. Results Serum sICAM-1 levels were significantly elevated in patients with AMI or UAP compared with that of control group, while higher hsCRP level was observed only in the patients with AMI compared with those with UAP and the control group. By linear regression analysis, only serum sICAM-1 levels were correlated with coronary risk score (r=0.445, P
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Aim To explore the influence of atorvastatin on the ultramicrostructure of the membrane surface of the rabbit endothelial cells in rabbit atherosclerosis(AS)in the nanometer level.Methods A total of 44 male New Zealand white rabbits were randomly divided into 3 groups:control group consisting of 12 rabbits,AS group consisting of 16 rabbits and atorvastatin group consisting of 32 rabbits.By the end of 2nd,6th week 6~8 rabbits of each group were sacrificed and the middle segments of thoracic aortas were obtained to be observed with atomic force microscope.Results The control group vascular endothelial cells(VECs)were fusiform in shape and aligned regularly.Their size were about 11.96 ?m?3.72 ?m and their macroaxis were in parallel with the direction of hemokinesis.VECs in the atherosclerotic group were in deformity and bigger than those of the control group.They aligned irregularly and their volumes changed to be swelled.The membrane protein of VECs in the control group was composed of many round and elliptical eminences,which were almost in the same size.and with distinct boundary lines.The membrane protein of VECs in the atherosclerosis group was composed of many irregular eminences in different size.It was vague among the eminences in which there were many holes.But the VECs of atorvastatin group were better than those of atherosclerosis group.The ultramicrostructure of the membrane surface of the atorvastatin group VECs was obviously improved.Meanwhile,the mean roughness(Ra)of membrane protein of three groups was compared.The Ra of the atherosclerosis group was significantly higher than that of the control group and the atorvastatin group(P