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To clone human mitogen-activated protein kinase kinase 6 (MKK6) gene promoter and explore its transcription activity by ubiquitin specific peptidase 22 (USP22). Methods: MKK6 gene promoter was amplified by PCR and two bases mutation within USP22 binding site was subsequently introduced. The wild type and mutant MKK6 promoter were inserted into the luciferase report vector pGL3-Basic, respectively. Recombinant plasmids were co-transfected with plasmid pRL-TK into HeLa cells, and the luciferase activities were measured by dual luciferase reporter system. Furthermore, the direct interaction between USP22 and MKK6 promoter was detected by chromatin immunoprecipitation (ChIP) assay. Finally, the MKK6 transcription activity was measured after knockdown of USP22. Results: The recombinant luciferase report vectors containing wild or mutant type of MKK6 promoter were successfully constructed. Mutation of USP22 binding site resulted in decrease of MKK6 promoter-driven luciferase activity in HeLa cells (P<0.05). USP22 could interact directly with MKK6 promoter. Down-regulation of USP22 led to the decreased MKK6 mRNA expression (P<0.05). Conclusion: USP22 could regulate the transcription activity of MKK6 gene in HeLa cells.
Subject(s)
Humans , HeLa Cells , Luciferases , MAP Kinase Kinase 6 , Promoter Regions, Genetic , Thiolester Hydrolases , Metabolism , Transcription, GeneticABSTRACT
Objective To explore the timing of surgical operation and surgical incision for the treatment of head of electrical injury and scarred alopecia with soft tissue expander .Methods Twenty-three patients with high tension electrical injury and scarred alopecia were collected ,the soft tissue expander was used to repair the wound and scar area.Selected the expander to embed the incision:patients with scarred alopecia were selected to be near the hair area in the scar;high tension electrical injury in patients with incision location selection based on the time of injury and wound repair requirements vary;within 5 days after injury ,surgery incision can be selected from the wound about 2-3cm;more than 5 days after injury,the patients chose to stay away from the wound incision ,incision about 4-5cm. Results Of 23 patients,postoperative scalp expansion flap good blood supply ,a period to solve the high tension electrical injury caused by skull necrosis and scarred alopecia problems .The patients according to the wound debride-ment and repair requirements using different surgical incision for dilator implantation ,wound and flap infection ,dilated flap poor blood supply ,necrosis,dilator exposed and other issues did not occur after surgery .Conclusion Flexible use of expander embedded surgical incision can be more effectively adapted to the wound repair requirements ,timely reduction of implantation expander surgery when the difficulty of stripping ,shorten the operation time,the maximum possible protection of healthy scalp tissue ,and eventually get a satisfactory clinical cosmetic repair effect .
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OBJECTIVE:To investigate the anticoagulation effects of warfarin on the lower extremity deep venous thrombosis (LEDVT),and to analyze its influential factors,in order to provide scientific basis for individualized medication of warfarin in the clinic. METHODS:Totally 140 cases of LEDVT were selected and treated with warfarin on the and day after admission with initial dose of 5.0 mg,qd,2.5 mg for following 2 days,orally,qd. The dose of warfarin was adjusted 72 h after medieation according to INR of patients. The dose of warfarin wasused as the maintenance dose when INR reached the anticoagulant target. Clinical data of patients were recorded,and blood biochemical indexes and coagulation function were detected. The influential factors of anticoagu-lation effects were analyzed by multiple-linear regression. At the same time,the results of INR were recorded before medication and 24,48,72 h after medication. RESULTS:INR of patients receiving warfarin had the potential to increase,compared to before medication;24,48,72 h after medication,INR value showed a gradual upward trend,without statistical significance(P>0.05). The change of INR within 24 h after medication was less than that within 24-48 h after medication;the change of INR within 24-48 h after medication was less than that within 48-72 h after medication,with statistical significance (P<0.01). The influence of various factors on the anticoagulant effect of warfarin in descending order was as follows:age,weight,low density lipoprotein cholesterol,plasma albumin,disease duration. Among them,age and low density lipoprotein cholesterol were positively correlated with anticoagulation,while body weight,plasma albumin and disease duration were negatively correlated with anticoagulation. CONCLUSIONS:Both age and body weight are the main influential factors for anticoagulation effect of warfarin. Individualized medication should be implemented in order to improve the anticoagulation effects of warfarin.
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Objective To investigate the clinical effect of large piece of mid -thick head skin applicated in repairing of wound in childrens′functional location.Methods 24 children with functional parts burn and scar con-tracture were selected.Using electric machine,the large piece of mid -thick head skin(thickness ≥0.25mm)was taken,then transplanted it to functional location directly or punching,and dressed or suture package fixed it,1 week after the skin transplanting,anti -scarring therapy and functional exercise were carried out.Control group:selected the thighs,back,abdomen as donor sites;The scar of operation area of the two groups were evaluated and analyed after 3 months by the Vancouver Scar Scale Rating.Results In the study group,the functional parts burn and scar con-tracture deformity was primary wound repairing,by the early anti -scarring therapy,the function and appearance was satisfied with repairing,during follow -up,there were varying degrees of repair site scar,but functional activity was not significantly affected;There were no scar hyperplasia of the head blood supply area,no bald hair deformity,compared with the control group after 3 months:surgical color (t =0.48,P >0.05),surgical vascular distribution (t =0.47, P >0.05),scar thickness (t =0.07,P >0.05)and softness (t =0.42,P >0.05),four indicators had no significant statistical difference.Conclusion The large piece of mid -thick head skin transplanting applied in children functional parts and scar contracture deformity restorative treatment,combined with early anti -scarring therapy and functional exercise,function and appearance can achieve satisfactory clinical results,but also for further post -repair function and appearance of the skin to provide more options source and surgical methods.
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ZnO nanotubes were prepared via electrospinning the Zn ( AC ) 2-polyacrylonitrile-polyvinylpyrrolidone ( PAN-PVP) precursor, followed by thermal decomposition of the above polymers from the precursor fibers. SEM and XRD characterization confirmed that the as-prepared ZnO nanofibers presented the hollow nanotube form, which was composed of ZnO nanoparticles with the size of about 40 nm in wurtzite crystal structure. By mixing with graphene, the obtained ZnO-graphene composite modified glassy carbon electrode ( ZnO-RGO/GCE ) was successfully constructed by dip-coating, which was used for the determination of Pb2+in water. With the sensitive response of the ZnO-RGO/GC electrode to Pb2+in solution was demonstrated by square wave stripping voltammetry, the response pctential was at -0. 4V. Under the optimized conditions, a good linear relationship between peak current and Pb2+ concentration was obtained in the range of 2. 4×10-9-4. 8×10-7 mol/L (R=0. 9970) by 10 min preconcentration at -1. 0 V in 0. 1 mol/L HAc-NaAc buffer solution (pH=4. 6). The detection limit was 4. 8×10-10 mol/L (S/N>3). The ZnO-RGO/GC electrode had good stability. The practical analytical application of the ZnO-RGO modified electrode was assessed by the measurement of the actual water sample and the result was consistent with that obtained by ICP-MS.
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ZnO nanoparticle-containing carbon composite nanofiber ( ZnO-CNF ) was prepared by the electrospinning of the ZnCl2-PAN precursor, followed by preoxidation and carbonization. The ZnO nanoparticles were uniformly distributed on the surface of the carbon nanofiber with the size of 20-30 nm, confirmed by scanning electron microscopy ( SEM ) . The wettability of the ZnO-CNF was studied by water contact angle test. With Nafion as an additive, the ZnO-CNF modified electrode was successfully constructed by dip-coating. The surface morphology and electrochemical properties of the modified electrode were investigated by SEM and cyclic voltammetry. There was a sensitive response of the ZnO-CNF modified electrode on Pb ions in solution, demonstrated by square wave stripping voltammetry. Under the optimized conditions, a good linear relationship between peak current and Pb2+concentration was obtained in the range of 2. 4×10-10-2. 4×10-7 mol/L (R=0. 998) by 10 min preconcentration at -1. 0 V in 0. 1 mol/L NaAc buffer solution (pH=4. 6). The detection limit was 4. 8×10-11 mol/L. The practical analytical application of the ZnO-CNF modified electrode was assessed by the measurement of the actual water sample and the result was consistent with that obtained by ICP-MS.
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OBJECTIVE@#To clone 5' untranslated region of human IPO8 gene and determine its transcription activity.@*METHODS@#We used 5' rapid amplification of cDNA ends (RACE) analysis to identify the IPO8 transcription start site (TSS), and amplified series truncated 5' UTR fragment containing transcription start site. The PCR productions were inserted into luciferase report vector pGL3- Basic. After confirmation by restriction enzyme digestion, the recombinant plasmids were cotransfected into Saos-2 cells with plasmid pRL-TK. The luciferase activities were measured by dual luciferase reporter system.@*RESULTS@#The IPO8 gene transcription start site was established. The electrophoresis analysis of restriction enzyme digestion and DNA sequencing verified the fragments were successfully amplified and inserted into pGL3-Basic. After the recombinant plasmids transfected, the highexpressions of luciferase were detected in Saos-2 cells.@*CONCLUSION@#The recombinant vector containing IPO8 promoter is constructed successfully, which provides a foundation for determining expressional regulation of IPO8 in the further study.
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Humans , Cloning, Molecular , DNA, Complementary , Genetic Vectors , Luciferases , Plasmids , Promoter Regions, Genetic , Transfection , beta Karyopherins , GeneticsABSTRACT
The hydrophilicity of the normal decoction pieces (NDP) of Indigo Naturalis is not good, therefore, it is not suit for decoctions. In this paper, powder modification technology is used and some NDP and alcohol are ground together in the vibromill to prepare the hydrophilic decoction pieces (HDP) of Indigo Naturalis. Initially, the properties of NDP, ultrafine decoction pieces (UDP) and HDP are compared, the hydrophilicity of UDP was promoted slightly, that of HDP is promoted dramatically. Then, three batches of Indigo Naturalis are prepared to HDP separately, but there is no obvious difference in the contact angle. Furthermore, the size distribution, surface area and micro-shape of HDP are bigger than that of UDP and smaller than NDP. The contents of indigo and indirubin in three decoction pieces are the same, as well as the species of inorganic substance, although there is a little difference in the proportion of five inorganic substances. The fact suggests the change of physical state and the qualitative and quantitative change of organism and inorganic substances are not the main factors to influence the hydrophilicity. In addition, hydroxyl, methylene and methyl can be identified at the wavenumber of 3 356 cm(-1) and 1 461 cm(-1) in infrared spectrum; the content of alcohol in HDP is 0.67% measured by gas chromatogram. The stability of HDP in the heating condition is studied, the fact suggests the hydrophilic effect of HDP at 40 degrees C is relatively stable. All above research suggests that the alcohol is the main factor to influence the hydrophilicity and maybe the intermolecular force which fixed alcohol molecule on the surface of Indigo Naturalis is the basic principle to produce the hydrophilicity.
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ObjectiveTo investigate the impact of blood glucose,blood pressure and blood uric acid level on hematuria in patients with acute coronary syndrome (ACS).MethodsOne hundred and sixty-two ACS patients were selected and received standardized treatment after admission to hospital.Urine test was taken and patients were divided into no hematuria group(37 cases),microscopic hematuria group (56 cases) and gross hematuria group(69 cases) according to the results.Blood pressure,fasting and postprandial 2 hours blood glucose,glycated hemoglobin and blood uric acid level were measured and compared among three groups.ResultsMicroscopic hematuria group compared with no hematuria group,fasting and postprandial 2 hours blood glucose,glycated hemoglobin,systolic blood pressure and blood uric acid level raised 58.0％[(7.9 ±0.7) mmol/L vs. (5.0 ± 1.1) mmol/L],33.3％[(12.4 ±0.8) mmol/L vs.(9.3 ± 0.6 ) mmol/L ],48.2％ [ ( 8.3 ± 0.8 )％ vs.( 5.6 ± 0.5 )％ ],23.8％ [ ( 151.6 ± 7.0) mm Hg ( 1 mm Hg =0.133 kPa) vs.(122.5 ±9.9) mm Hg],29.2％ [(635.4 ±47.4) μmol/L vs.(491.8 ±83.4)μmol/L]respectively,there were significant differences (P ＜ 0.05 or ＜ 0.01 ).Gross hematuria group compared with microscopic hematuria group,the above mentioned indexes raised 16.5％[ (9.2 ± 1.1 ) mmol/L vs.(7.9 ± 0.7)mmol/L],30.6％[ ( 16.2 ± 1.8) mmol/L vs.( 12.4 ± 0.8) mmol/L],14.5％[ (9.5 ± 0.8)％ vs.(8.3 ± 0.8)％ ],18.8％[(180.1 ± 12.3) mm Hg vs.(151.6 ±7.0) mm Hg],34.6％[(855.5 ±74.5) μ mol/L vs.(635.4 ±47.4 ) μ mol/L ] respectively,there were significant differences ( P ＜ 0.05 or ＜ 0.01 ).Gross hematuria group compared with no hematuria group,the above mentioned indexes increased significantly(P＜ 0.01 ).The level of diastolic blood pressureamong three groups had no significant difference(P ＞ 0.05).The Logistic regression analysis showed that fasting and postprandial 2 hours blood glucose (r =3.175,P =0.001 ;r =0.906,P =0.001 ),glycated hemoglobin ( r =16.109,P =0.001 ),systolic blood pressure (r =0.429,P =0.003 ),blood uric acid level(r =1.317,P =0.004) were risk factors on hematuria after antiplatelet and anticoagulant therapy in ACS patients,the impact of glycated hemoglobin and fasting blood glucose on hematuria was stronger than that of blood uric acid level and systolic blood pressure.ConclusionWith the increase of blood glucose,systolic blood pressure,blood uric acid,the risk of hematuria increases in ACS patients.
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Objective To investigate the relationship between the activity of nicotinamide adenosine dinucleotide phosphate(NADPH ) oxidase in peripheral blood mononuclear cell and left ventricular remodeling in patients with acute myocardial infarction (AMI).Methods Fifty-two patients with AMI hospitalized from December 2010 to June 2011 were selected as AMI group,and 25 patients with paroxysmal supraventricular tachycardia hospitalized in the same period were selected as control group.All patients were examined by Doppler echocardiography and left ventricular mass index (LVMI) was calculated.Fasting blood was drawn on the second day after hospitalization to measure the level of biochemical indexes,malondialdehyde(MDA ) and the activity of NADPH oxidase.The correlation of the level of MDA,the activity of NADPH oxidase and LVMI was analyzed.Results The level of MDA in AMI group was significantly higher than that in control group [(6.14 ±t 2.80) μ mol/L vs(2.95 ± 1.82) μmol/L,P ＜ 0.01 ].The levd of O2- in AMI group was also obviously higher than that in control group [(21.61 ± 10.66) RLU/mg protein vs.( 7.12 ± 4.48 ) RLU/mg protein,P ＜ 0.01 ],as well as the activity of NADPH oxidase [ ( 2.84 ± 1.05 ) RLU/mgprotein vs.( 1.97 ± 0.80) RLU/mg protein,P ＜ 0.01].Positive correlation was found between LVMI and the level of MDA(r =0.57,P ＜ 0.01 ),as well as between LVMI and the activity of NADPH oxidase(r =0.71,P＜0.01 ).Conclusions The activity of NADPH oxidase in peripheral blood mononuclear cell inereases in AMI patients and plays an important role in left ventricular remodeling after AMI.