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Objective To observe the changes in forkhead/winged helix transcription factor p3(Foxp3), regulatory T cells(Treg),retinoid-related orphan receptor gamma(RORγt)in rat model of chronic obstructive pulmonary disease(COPD). Methods Twenty Sprague-Dawley(SD)rats were randomly divided into normal control group and COPD model group,with 10 rats in each group. The COPD model was reproduced by smoke inhalation and tracheal instillation of lipopolysaccharide(LPS),and no such treatment was conducted in normal control group. Twenty-eight days after the model reproduction,the pulmonary function was determined,the pathological changes of lung tissue were observed with haematoxylin-eosin(HE)staining,interleukins(IL-6,IL-10)in serum were detected by enzyme-linked immunosorbent assay(ELISA),CD4+CD25+Foxp3+Treg of peripheral blood was determined by flow cytometry,and the expressions of Foxp3,RORγt,IL-17 protein in lung tissue were assayed by Western Blot. Results Under light microscope,significal interstitial infiltration of inflammatory cells was found in alveoli and interstitial tissue of the lung,and destruction of alveolar tissue,alveolar wall thinning,and even rupture to fuse into bullae,and bleeding into alveoli in different degress could be observed. Compared with the normal control group,forced vital capacity(FVC),forced expiratory volume in 0.3 second(FEV0.3),FEV0.3/FVC,peak expiratory flow(PEF)in model group were significantly decreased〔FVC(mL):8.04±2.03 vs. 9.97±2.14,FEV0.3(mL):6.16±2.23 vs. 8.84±2.12,FEV0.3/FVC:0.70±0.09 vs. 0.85±0.11,PEF(mL/s):33.56±4.76 vs. 40.14±5.64, P<0.05 or P<0.01〕. Serum IL-6 was obviously increased(ng/L:93.17±20.96 vs. 76.28±13.24,P<0.05), IL-10 was significantly decreased(ng/L:78.62±15.17 vs. 104.34±19.46,P<0.01),and CD4+CD25+FoxP3+Treg was significantly diminished〔(2.75±0.83)% vs.(4.16±1.14)%,P<0.01〕in model group compared with those in the normal control group. The expression of Foxp3 protein in lung tissue in model group was significantly down-regulated compared with that in the normal control group(gray scale:0.38±0.15 vs. 0.63±0.11,P<0.01), and RORγt and IL-17 protein expressions were significantly up-regulated〔RORγt(gray scale):0.96±0.23 vs. 0.47±0.11,IL-17(gray scale):1.02±0.24 vs. 0.34±0.08,both P<0.01〕. Correlation analysis showed that FEV0.3 was positively correlated with Foxp3(r=0.585,P<0.05),and FEV0.3/FVC was negatively correlated with IL-6 and RORγt(r=-0.655,r=-0.607,both P<0.05). PEF was positively correlated with Treg(r=0.573, P<0.05),and negatively correlated with IL-17(r=-0.198,P<0.05). IL-6 was negatively correlated with Foxp3(r=-0.603,P<0.05),and positively correlated with RORγt(r=0.588,P<0.05). IL-10 was positively correlated with Treg(r=0.573,P<0.05). Treg was positively correlated with Foxp3(r=0.607,P<0.05), and negatively correlated with IL-17(r=-0.569,P<0.05). Foxp3 was negatively correlated with RORγt(r=-0.591, P<0.05). RORγt was positively correlated with IL-17(r=0.578,P<0.05). Conclusion There is a relationship among decreased pulmonary function,inflammation and imbalance of Foxp3/Treg and RORγt/Th17 in COPD.
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Objective To observe the effects of Liuwei Buqi capsules on CD4+CD25+forkhead box protein 3+(Foxp3+)regulatory T cells(Tregs),helper T cells(Ths)in patients with chronic obstructive pulmonary disease (COPD). Methods Eighty COPD patients admitted into the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine from January 2012 to May 2013 were randomly divided into a traditional Chinese medicine (TCM)experimental group(40 cases)and a western medicine control group(40 cases);there were 40 research volunteers enrolled as healthy controls. After admission,both the TCM experimental group and western medicine control group were given conventional western medicine comprehensive treatment. Additionally,in the TCM experimental group,Liuwei Buqi capsules were given orally once 3 pills(0.4 g/pill)and twice a day,while in the western medicine control group nifedipine was given,once 10 mg and 3 times a day,30 days constituting one therapeutic course for both groups. BODE evaluation means that the body mass index(B),the degree of airflow obstruction(O),the scores of dyspnea(D)and excercise capacity(E)of COPD patients were evaluated. A spirometer was used to observe the changes in the patient's pulmonary function to detect the dyspnea score. The γ-interferon(IFN-γ),interleukin (IL-4 and IL-17)in serum were detected by enzyme-linked immunosorbent assay(ELISA). The expression of Treg of peripheral blood was detected by flow cytometry. Results Compared with healthy control group,lung function parameters,IL-4,CD4+CD25+Treg and CD4+CD25+Foxp3+Treg of peripheral blood were increased,while dyspnea score,BODE score,the expressions of IFN-γ,Th1/Th2,IL-17 in serum were decreased significantly in the COPD group(P<0.05 or P<0.01). After treatment,compared with those before treatment in the two groups, the forced expiratory volume in 1 second(FEV1),FEV1/forced vital capacity(FVC),IL-4,CD4+CD25+Treg, CD4+CD25+Foxp3+ Treg were all increased,while dyspnea score,BODE score,IFN-γ,Th1/Th2,IL-17 were all decreased(P<0.05 or P<0.01),and the peak expiratory flow(PEF)was decreased in western medicine control group and increased in TCM experimental group. After treatment,the comparisons of ratio of FEV1/FVC〔(78.12±14.96)%vs.(67.52±10.39)%〕,BODE score(1.07±0.72 vs. 1.77±0.74),IL-17(μg/L:40.80±8.97 vs. 48.22±6.51) Th1/Th2(1.05±0.23 vs. 1.42±0.21)and CD4+CD25+Foxp3+ Treg〔(6.61±2.26)% vs.(5.25±2.03)%〕between the two groups were all of statistical significant difference(all P<0.05). Conclusion Liuwei Buqi capsules can improve COPD symptoms by up-regulating the expressions of CD4+CD25+Foxp3+ Treg,IL-4,and down-regulating the expressions of IFN-γ,IL-17 to correct the balance of Th1/Th2 in patients with COPD.
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AIM:To explore the fingerprint method for quality control of Danhong sterile powder for injection (Radix et Rhizoma salviae mihiorrhizae,Flos carthami).METHODS:The samples were determined by Agilent 1100 DAD-HPLC with SinoChrom ODS-BP column(250 mm×4.6 mm,5 μm),by gradient elution using methanol-0.1% phosphoric acid,as mobile phase 30 ℃ column temperature.flow rate of 1.0 mL/min,detection wavelength was set at 280 nm,and inject volume 20 μL.RESULTS:Its fingerprint exhibited better separation for each peak,revealing amount of finger information,Based on the ten batches of Danhong sterile powder for injection,according to the technical requirements of fingerprint on injection of Chinese traditional medicine.CONCLUSION:Proposed HPLC fingerprint can be used as the important evidences for the quality control of Danhong Injection.
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The acquisition of heart rate variability (HRV) signals is very important to physiological research and clinical diagnosis. In order to ensure the accuracy of acquired heart rate variability signals, we must consider the method of acquisition of heart rate variability signals. By using the relation between the signal singularity and its wavelet transforms, a software for R wave in detection is designed. The result is very satisfying in detecting R wave of ECG database of MIT/BIH.
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Humans , Algorithms , Electrocardiography , Heart Rate , Physiology , Signal Processing, Computer-Assisted , SoftwareABSTRACT
Objective To investigate the expression of focal adhesion kinase in peripheral blood of colorectal cancer and its clinical significance.Methods Focal adhesion kinase in peripheral blood was determined with flow cytometry in 40 cases of colorectal cancer and 20 cases of control group.Results The positive rate of focal adhesion kinase in colorectal cancer group was 60.0% and in control group was 25.0%,respectively(P
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AIM:To explore the fingerprint method for quality control of Danhong sterile powder for injection(Radix et Rhizoma salviae miltiorrhizae,Flos carthami).METHODS:The samples were determined by Agilent 1100 DAD-HPLC with SinoChrom ODS-BP column(250 mm?4.6 mm,5 ?m),by gradient elution using methanol-0.1% phosphoric acid,as mobile phase 30 ℃ column temperature,flow rate of 1.0 mL/min,detection wavelength was set at 280 nm,and inject volume 20 ?L.RESULTS:Its fingerprint exhibited better separation for each peak,revealing amount of finger information,Based on the ten batches of Danhong sterile powder for injection,according to the technical requirements of fingerprint on injection of Chinese traditional medicine.CONCLUSION:Proposed HPLC fingerprint can be used as the important evidences for the quality control of Danhong Injection.
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The total proteins of plain E.coli(DH5 ?)were coupled to the Sephacryl S300 oxidized with periodate,and the preliminarily purified antiserum was loaded repeatedly on the column for 18 hours.The unspecific anti\|E.coli antibodies would be absorbed and the antibodies would be purified.It was more specific to do western blot using antibodies treated by this method than using ones without treatment.