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 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.

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.

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 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 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 investigate the efficacy of percutaneous transhepatic variceal embolization (PTVE) with Cyanoacrylate (TH glue) in treating large gastric fundal variceal bleeding.Methods PTVE was performed on 24 patients with TH glue injected into the main stem of left gastric vein and its fundic branches.The degree of varices in gastric fundus,rebleeding rate and survival rate after the procedure were compared with those before.Results Varices in gastric fundus were all embolized successfully with TH glue.The diameter of varices was reduced to below 5mm or disappeared in 20 patients (83.3％),and reduced to 5-10mm in the other 4 ( 16.7％ ) During the follow-up period of 3-36 months(mean 16.6 months),the rebleeding rate and mortality were 12.5 ％ ( 3/24 ),and 12.5 ％ (3/24),respectively.One patient died of liver cancer,and two others died of chronic liver failure.Conclusion PTVE with TH glue is of ideal therapeutic effect to block the feeding veins of large gastric fundal varices.

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

BACKGROUND: Studies have demonstrated that exogenous neural stem calls (NSCs) could repair nerve and promote recovery of neurofunction following cerebral hemorrhage. However, the influence of internal environment after cerebral hemorrhage on the survival and differentiation of NSCs is a complex and variable process.OBJECTIVE: To observe the survival and differentiation of human embryonic NSCs implanted in rats with cerebra hemorrhage.DESIGN, TIME AND SETTING: Open, immunohistochemistry, experiment was performed at the Laboratory of Molecular Biology, Affiliated Hospital of Luzhou Medical College from May 2007 to April 2008.MATERIALS: A total of 40 female SD rats were provided by the Experimental Animal Institute, Chinese Academy of Medical Sciences. Brain of 8-week aborted fetus was obtained from Department of Gynaecology and Obstetrics, the People's Hospital of Deyang City.METHODS: Cerebral cortex cells of 8-week aborted human fetus were harvested and cultured in vitro to obtain human embryonic NSCs. Cerebral hemorrhage rat models were established via injection of autologous arterial blood in caudate nucleus. Two days after modeling, 5 μL BrdU-labeled human embryonic NSCs suspension was transplanted at four points surrounding hematoma cavity in the rats. After 1 and 2 weeks, rats were sacrificed. Adjacent sections were doubly stained by BrdUImicrotubule-associated protein 2 (MAP-2) and BrdU/glial fibrillary acidic protein (GFAP).MAIN OUTCOME MEASURES: The survival, immigration and differentiation of human embryonic NSOs implanted in rats were observed by immunohistochemistry and immunofluorescance staining.RESULTS: BrdU-positive cells were oval and brown. At 1 and 2 weeks after implantation, BrdU-positive calls survived and migrated, and they migrated more widely at 2 weeks after implantation. At 1 week after implantation, BrdU/MAP-2-positive cells and BrdU/GFAP-positive calls were observed in cerebral tissue sections, and the number of BrdU/MAP-2-positive cells was more than BrdU/GFAP-positive calls; At 2 weeks after implantation, BrdU-positive cells found in choroid plexus and blood capillary were significantly reduced, and BrdU/GFAP-positive calls were more than BrdU/MAP-2- positive cells.CONCLUSION: Implanted human embryonic NSCs can survive and migrate in the hemorrhage region, gradually differentiate into neurons or astrocytes.

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.