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.

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.

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.

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

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.

Objective To investigate the effect of electroacupuncture on expression of growth associated protein-43 (GAP-43) and Nogo-A in brain after focal cerebral ischemia/reperfusion in rats. Methods 48 male Sprague-Dawley rats were randomly divided into sham group, model group and electroacupuncture group, and the latter two were modeled as middle cerebral artery occlusion and reperfusion with nylon monofilament suture. Electroacupuncture was performed 90 min after modeling in the electroacupucture group at acupoints of Neiguan (PC06), Shuigou (DU26), Sanyinjiao (SP06), Baihui (DU20), for 30 min, once a day. The sham group and the model group were conventionally fed in cages without any intervention. 8 rats of each group were assessed with Longa's score, and the expressions of GAP-43 and Nogo-A were detected with immunohistochemistry 7 d and 14 d after modeling respectively. Results The sham group presented no neurological symptoms. There was not different in Longa's score between the model group and the electroacupuncture group 7 d after modeling (P>0.05), but was different 14 d after modeling (P<0.05). GAP-43 positive cells was not found in the sham group, but could be found around cerebral ischemia 7 d after modeling, which decreased 14 d after modeling in the model group. GAP-43 positive cells increased significantly in the electroacupuncture group compared with the model group at each time (P<0.01). Nogo-A positive cells was few found in the sham group, and was more in the model group (P<0.01). The expression of Nogo-A decreased significantly in the electroacupuncture group compared with the model group at each time (P<0.01). Conclusion Electroacupuncture can improve neurological function of focal cerebral ischemia/reperfusion rats, which may be associated with the increase of GAP-43 and descrease of Nogo-A in peri-infarct regions

Objective To investigate the influence on biological characteristics in human pancreatic cancer cells after beding transfected by two anti-carcinoma miRNAs at the same time.Methods Pancreatic cancer cells PANC1,SW1990 and normal pancreatic cells AR42J were transfected by miR-34a and(or) miR-let7 by liposome.Cells transfected with negative control miRNA (miR-NC) and untransfected were as controls.The expression of miR-34a and miR-let7 were detected by real-time fluorescent quantitative RT-PCR.The cell proliferation was detected by MTT test and the migration and invasion were evaluated by transwell assay.The apoptosis rate was measured by flow cytometric analysis.Results After being transfected with miRNAs,the expression of miR-34a and miR-let7 in double transfection group (miR-34a and miR-let7 were transfected at the same time),miR-34a transfection group,miR-let7 transfection group was significantly up-regulated than those in miRNA-NC transfection group and untransfected group in PANC1 cells,SW1990 cells and AR42J cells,repectively.The difference which was statistically significant (P ＜0.05)indicating that cells were successfully transfected.The cell proliferation in double transfection group of PANC1 cells and SW1990 cells were (0.665 ± 0.01,0.6375 ± 0.03),which were significantly inhibited compared with (0.974 ± 0.03,0.971 ±0.05) in miR-NC group and (0.8875 ±0.05,0.8625 ±0.06) in miR-let7 group.The difference was statistically significant(P ＜ 0.05).The cell proliferation activity in double transfection group was lower than those in miR-34a group (0.795 ±0.06,0.7925 ±0.06),but did not have statistically significant difference.There was no significant change in AR42J cells.Cell invasion assay showed that the number of PANC1 cells permeating substrate membrane in miR34a group (103.7 ± 3.28) and miR-let7 group (100.7 ± 1.76) were significantly fewer than miR-NC group (231.3 ±2.6) and untransfected group (153.7 ±2.6).The number of cells permeating substrate membrane in double transfection group(61.67 ± 3.18)was fewer than miR-34a group and miR-let7 group,respectively.The difference was statistically significant (P ＜ 0.01).The migration test had consistent results with invasion test.The changes of invasion and migration in SW1990 cells were similar to those in PANC1 cells.The apoptosis rate of PANC1 cells in miR-34a group,miR-let7 group,double transfection group,miR-NC group and untransfected group was (16.66 ± 1.27) ％,(15.46 ± 0.33) ％,(23.35 ± 1.80) ％,(9.33 ± 0.31) ％ and (8.83 ± 0.36) ％ respectively.Single transfection group had higher apoptosis rate than miR-NC group and untransfected group (P ＜0.05).Double transfection group had a significantly higher apoptosis rate than miR-let7 group (P ＜ 0.05),while there was no significant difference between double transfection group and miR-34a group.Conclusions The cell proliferation,invasion and migration in double miRNAs transfected pancreatic cancer cells were significantly down-regulated compared with those in single miRNA transfected cells,while apoptosis rate in double miRNAs transfection group was higher than single miRNA transfection group.Thus,the combination of two anti-cancer miRNAs may exert a more significant synergistic antitumor effect.

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.

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.
Algorithms ; Electrocardiography ; Heart Rate ; physiology ; Humans ; Signal Processing, Computer-Assisted ; Software

The fourth time national survey of Chinese materia medica (CMM) was used. The quadrat survey contained recording on species and amounts of plants, collection of specimens and CMM. The weight of main medicinal plant was measured. The quadrat numbers were calculated on regions, plant species and sea level of samples from different regions. There were 703 species of medicinal plants. The 143 species of main medicinal plants, which occupied to 74.5% in main medicinal plants in Chongqing, belonged to 71 families and 132 genera. There were 128 species of wild main medicinal plants. The 30 species of main medicinal plants were cultivated (including 15 species of both wild and cultivated medicinal plants). Among them, 14 species were in large-scale cultivation. This investigation helped the understanding of present situation of medicinal species distribution in Chengkou County, the habitats of wild medicinal plants, and the resources reserves, which provided references for the continuous development of Chinese herbal medicine resources.