In order to obtain the bifunctional chimeric molecule of single-chain urokinase-type plasminogen activator (scu-PA) which can inhibit platelet aggregation, PRGDWR peptide was inserted into the site between Gly 118 and Leu119 (called insertion mutant B, InB). The recombinant gene of InB was expressed by Pichia pastoris. The secreted protein was purified by metal chelate affinity and strong cation exchange chromatography. The amidolytic ability of mutant InB is 5 900 IU/mg, the kinetic constants is: KInB m,plg=56.8 μmol*L-1，kInBcat,plg=0.33 s- 1. The kinetic constants of plasminogen activation reaction is: KInB m,plg=0.397 μmol*L-1，kInBcat,plg=0.0164 s-1. Fibrin inhibit the catalytiv ability of InB during plasminogen activatio n, the influence factor is 0.463(means InB remain 46.3% of the catalytic abili ty when fibrin was involved in the reaction system). The mutant not only has alm ost the same catalytic ability as wild type scu-PA, but also has strong ability of anti-platelet aggregation(compared with scu-PA), IC50 of InB is 12.7 μmol*L-1.

Because the influence of fibrin on the reaction of plasm inogen activation by various plasminogen activators is different, the kinetic co nstant of the reaction of plasminogen activation catalyzed by InB with and witho ut fibrin were detected. The result is: Kfibrinm＝4.2 μmol*L -1，greater than the normal Km=0.379 μmol*L-1; kfib rincat＝0.107 s-1，greater than the normal kcat=0.0165 s-1. The results suggest that existence of fibrin in the reaction system of plasminogen activation depress the affinity between InB and plasminog en, but accelerates the hydrolysis of plasminogen by InB. The count up effect is inhibition.

Objective To analyze the clinical characteristics of children infecting with Epstein -Barr virus. And in purpose to improve the knowledge of children infecting with Epstein -Barr virus.Methods We analyzed retrospectively to 97 cases of children which serodiagnosis of Epstein -Barr virus infectius in the aspects of the onset season,the age,clinical characteristics.And compared with complication group and no complication group in the aspects of the white cell count,neutrophils to white cell count ratio,lymphocyte to white cell count ratio and C -reactive protein.And contrasted the presence of statistical difference.Results The peak months were spring and summer(58 cases);Pre -school age was the peak age of EBV infectious(41 cases),and the rest respectively were toddlers age(33 cases),school age and youth(20 cases)and infant period(3 cases).The most frequently clinical manifestation respectively was pharyngitis,and other such as fever,lymphadenopathy,hepatosplenomegaly.There had no statistical difference in the two groups of complication group and no complication group in the aspects of the white cell count,neutrophils to white cell count ratio,lymphocyte to white cell count ratio and C -reactive protein(P >0.05).Conclusion The infant can suffer from infectious mononucleosis infect with EBV.Infectious mononucleosis have a variety of clinical manifestation,which always have some complications.The diognosis should combinate clini-cal manifestation with laberatory test.

Objective The purpose of this study was to investigate the effect of pinacidil, an ATP-sensitive K + channel opener in protecting myocardium of immature rabbit heart from ischemia-reperfusion injury.Methods Fifty-two rabbits of 3-4 weeks old, weighing 350-450g were anesthetized with intraperitoneal pentobarbital sodium(50mg/kg). Hearts were excised and connected to modified Langendorff perfusion system and passively perfused with Krebs-Henseleit buffer(KHB) at 76 cmH 2O. Hearts were made globally ischemic for 30 min by aortic crossclamping and then reperfused for 30min. During ischemia the hearts received 20-25 ml cardioplegic solution of diferent constituents every 15 min. There were four groups with 13 hearts in each group based on the diferent constituents of cardioplegic solution: group Ⅰ(control, no cardioplegic solution); group Ⅱ(KHB+K + 16mmol/l); group Ⅲ(KHB+K +16mmol/L+pinacidil 50?mol/L); group Ⅳ (KHB+K +16mmol/L+glibenclamide 10?mol/L). Left ventricle developed pressure (LVDP), left ventricle diastolic pressure (LVEDP),?dp/dtmax, coronary flow (CF), and the levels of CK, LDH and AST in coronary sirnus veneous effluent and by myocardial ultrastructural changes. Myocardial ultrastructure was examined before and after ischemia.Results Before myocardial ischemia, there were no significant differences in the above mentioned parameters among four groups. Group Ⅲ was the best in terms of postischemic recovery of LVDP, LVEDP,?dp/dtmax and CF. The changes in CK,LDH,AST levels and myocardial ultrastructure were the least in group Ⅲ.Conclusions Pinacidil can afford best myocardial protection against ischemia-reperfusion injury in immature rabbit hearts.

Objective:To evaluate the inhibitory effect of altered HA308-317 peptides on HLA-DR4 restricted CII specific T cell response.Methods:Three altered HA308-317 peptides and CII263-272 were synthesized using solid-phase techniques. The binding of altered HA308-317 peptides for HLA-DR4 molecules was assayed using flow cytometry. The suppressive effect of altered HA308-317 peptides on CII-mediated T cell proliferation was determined using 3H incorporation assay. The level of IL-2 in the supernatants was identified by ELISA. The expression of CD25 and CD69 on T cell surface were studied using flow cytometry.Results:The altered HA308-317 peptides were able to bind to HLA-DR4 molecules and competed with CII263-272. Altered HA308-317 peptides inhibited T cell proliferation and IL-2 production induced by CII263-272( P

Objective To evaluate the value of echocardiography in investigation the failure of interventional therapy of VSDs and to increase the successful rate. Methods 15 cases with failure of VSD closure through interventional approach were undertaken measurement of major parameters of the defect on left ventriculography and then followed by Philips 5500 color Doppler US for repeated multi-direction tangential measuring of the size, morphologic change, relation with peripheral structure and individual valvular regurgitation. Results (1) Marginal membranous VSD 10 cases; membranance aneurysm 8 cases, ≥2 outlets 6 cases. Width of basal part of membranous aneurysm was 8 ～ 18 (10 ?2) mm with depth of 3 ～ 10 (6 ? 2) mm, distance from aortic valve was 0 ～ 6 (2 ? 1) mm and from tricuspid valve was 2 ～ 5 (2 ?1) mm; associated with slight tricuspid valvular regurgitation. Aortic valvular regurgitation 5 cases showed slight to mild amount and without membranous aneurysm occurred in 2 cases. (2) Intracristal VSD 5 cases showed the size of defect as 6 ～ 10 (7 ? 1) mm, with distances of 2 ～ 3 (1 ? 0.8) mm from pulmonary valve and 1 ～ 2 (1 ? 0.6) mm from aortic valve associated with slight regurgitation in 4 cases, slight tricuspid valvular regurgitation in 5 cases and aortic valvular prolapse in 4 cases. Conclusions Many factors can influence the direction tangential continuous scanning should be taken for accurate localization and measurement of VSD in order to select the very sight occludor providing high successful rate of occlusion and long term efficacy.

Chronic Disease ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; therapeutic use ; Humans ; Male ; Prostatitis ; drug therapy ; Randomized Controlled Trials as Topic ; Suppositories

In order to investigate the comprehensive quality differences of the seeds harvested in different growing time, and offer theoretical guide for the optimization of seeds' production technology, we analyzed the apparent size, 1 000-grain weight, water absorbency, germination indexes, postharvest embryo rate change, relatively contents of coumarins and the yield of single plant of its seeds of different harvesting time, and compared their comprehensive quality by Topsis analysis. The results showed that there existed obvious differences in yield and quality between seeds of 3 harvesting times. While the harvesting time postponed, the yield of single plant raised, and the shrunken seeds rate decreased, the quality of seedlings increased, while the contents of coumarins showed a steady increase, and the germination rate decreased. The comprehensive quality of the seeds harvested in the black ripe time rank the first place, followed by the brown ripe time and the yellow ripe time. As the harvesting time delays, the seeds' comprehensive quality increases, therefore, we could put off the seeds' harvesting time properly for the high efficient seed production.
Bupleurum ; growth & development ; metabolism ; physiology ; Germination ; Plant Proteins ; chemistry ; metabolism ; Quality Control ; Seeds ; growth & development ; metabolism ; physiology ; Solubility ; Time Factors ; Water ; metabolism

OBJECTIVETo evaluate the efficacy and safety of Chinese herbal medicine combined with systemic chemotherapy and/or regional arterial perfusion for pancreatic cancer with liver metastases (PCLM).

METHODSWe retrospectively selected 292 patients with PCLM who were treated by Chinese herbal medicine combined with systemic chemotherapy and/or regional arterial perfusion at Tianjin Medical University Cancer Hospital from January 2001 to December 2010. All patients were assigned to the Western medicine treatment group (157 cases) and the integrative medicine treatment group (135 cases). Patients in the Western medicine treatment group were treated with gemcitabine (GEM)-based chemotherapy, and partial of them received regional arterial perfusion. Those in the integrative medicine treatment group additionally took Chinese herbs of clearing heat and eliminating mass for at least 4 weeks. The median survival time (MST) , adverse reactions and the incidence of complications were observed.

RESULTSThere was no statistical significance in general data between the two groups (P > 0.05). There was statistical difference in MST between the two groups (4.8 months vs 5.5 months, P < 0.05). No death occurred during chemotherapy or regional arterial perfusion. All toxic or adverse reactions were tolerable.

CONCLUSIONChinese herbal medicine combined with systemic chemotherapy and/or regional arterial perfusion was effective and safe, and it could be optimally selected as palliative therapy for PCLM.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Complementary Therapies ; methods ; Deoxycytidine ; analogs & derivatives ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Liver Neoplasms ; drug therapy ; secondary ; Pancreatic Neoplasms ; drug therapy ; pathology ; Retrospective Studies