Using ~(125)I-labeled anti-IL-2R_? McAb, we investigated the effect of rIL-2 R_? expression . In addition to increasing the IL-2 R_? expression the actived-lymphoblasts, we found that the higher concentration of IL-2 also directly induced the IL-2 R_? expression on the resting PBL, but the peak was at the 5th day. After treated by a higher concentration of IL-2, there was a positive interrelation of time courses between the incorporation of ~3H-TdR and/or ~3H-UR and IL-2 R_? level on resting PBL. This result indicated that it was by means of inducing IL-2 R_? expression that the higher concentration IL-2 caused the proliferation of the resting PBL.

By using 5 G 1-sepharose 4 B immune affinity column, we recovered approximately 160 ?g purified and soluble IL-2 receptor (sIL-2 R ) from 500ml PHA-stimulated human PBL supernatant. Only one band was showed in SDS-PAGE, its molecular weight was about 44 Kd. The purified sIL-2 R, which blocked the binding of anti-IL-2 R monoclonal antibody with IL- 2 R~+ cell, inhibited (at the higher doses) but enhanced (at the lower doses) the growth of IL-2 dependent cells.

Abstract Objective: To construct and express anti-IL-2R? genetically engineering antibody Fab and identify its biological activity.Methods: PCR were used to construct cloning vector 5G1-pA22 of anti-IL-2R? antibody Fab,by digestion of vector 5G1-pA22, anti-IL-2R? an-tibody Fab gene fragment was obtained and then this fragment was coupled with digesting product of pET28b plasmid by EcoRI and HindIII toget a recombinant expression vector 5G1-pET28b, it was highly expressed in the E. coli BL21(DE3) in the form of inclusion bodies, compoing20% of the total protein. After the renaturation of antibody Fab, its binding activity was identified by ELISA and antibody competitive inhibitionassay in vitro. Results: Anti-IL-2R? antibody Fab was successfully constructed and highly expressed in the form of inclusion bodies, its yield wasup to 1 mg/ml.When applied to ELISA, antibody Fab exhibited good spectfic reactivity with rIL-2R, antibody competitive inhibition assay invitro indicated that antibody Fab can compete with IL-2 for binding IL-2R in activated T cell surface.As a result,the activation and proliferationof T cells were inhibited and inhibition percentage exceeded 90%. Conclusion: Had successfully constructed and expressed anti-IL-2R? geneti-cally engineering antibody. Fab,it had good biological ativity.

Arsenicals ; pharmacology ; DNA Methylation ; drug effects ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; K562 Cells ; Membrane Proteins ; genetics ; metabolism ; Oxides ; pharmacology

Aim To study the anti-inflammatory effects and mechanisms of SBM isolated from Scutellariae radix by high hydrostatic pressure.Methods The effect of SBM on ConA-induced splenocyte proliferation and LPS-induced IL-1? production from peritoneal macrophage was studied in vitro,and the therapeutic effects of SBM on adjuvant-induced arthritis,formalin-induced paw oedema and acetic acid-induced vascular permeability were also investigated.Results SBM(31.25～500 mg?L-1)significantly inhibited ConA-induced splenocyte proliferation,IL-1? production from peritoneal macrophages in vitro.SBM 20 mg?kg-1 inhibited primary and secondary inflammatory reaction,decreased the elevated level of IL-1? released from peritoneal macrophages,inhibited splenocyte proliferation in adjuvant-induced arthritis in rats.Moreover,SBM inhibited formalin-induced paw oedema and acetic acid-induced vascular permeability.Conclusions SBM could inhibit inflammatory reaction.Its mechanisms might be related to the suppression of immune function and inhibition on proinflammatory cytokines production.

Aim To study the anti-inflammatory effects and mechanisms of SBM isolated from Scutellariae radix by high hydrostatic pressure.Methods The effect of SBM on ConA-induced splenocyte proliferation,LPS-induced PGE2 production and protein expression of COX-2 were studied in vitro,and the therapeutic effects of SBM on carragineen-induced paw oedema and acetic acid-induced twisting reaction were also investigated.Results SBM(15.63～250) mg?L-1 significantly inhibited LPS-induced PGE2 production and COX-2 enzyme activity in peritoneal macrophages in mice.SBM(125 mg?L-1)also inhibited ConA-induced protein phosphorylation of p38 MAP kinase and LPS-induced COX-2 protein expression.SBM(10,20 mg?kg-1)inhibited carragineen-induced paw oedema and acetic acid-induced twisting reaction.Conclusion SBM can inhibit inflammatory reaction.Its mechanisms may be related to the suppression of inflammatory medium production.

Objective To investigate the association between resting heart rate(RHR)and dyslipidemia. Methods A total of 772 subjects(average age:40. 1 ± 11.8 years;male:74. 7%)were recruited by random sampling method from medical checkup population. The survey data included resting heart rate,blood lipid and other metabolic parameters. The subjects were divided into three groups according to the RHR,as RHR ＜ 70 bpm,70 bpm ≤ RHR ＜ 80 bpm,RHR ≥ 80 bpm groups. Results RHR were positively correlated with triglyceides (TG),as well as total cholesterol(TC)(r = 0. 316 and r = 0. 110 respectively,all P value ＜ 0. 05). Low RHR had no significant correlation with HDL-C or LDL-C. The prevalences of hypertriglyceridemia and hypercholesterolemia increased significantly with RHR elevation(χ2 = 7. 214 and 5. 285 respectively,all P value ＜ 0. 05). By logistic regression analysis,adjusted for BMI,gender,serum creatinine,serum uric acid,fasting blood glucose and age,we found that compared with RHR ＜ 70 bpm group,the odds ratio(OR)values for hypertriglyceridemia was 1. 540 (95%CI:1. 086-2. 185,P=0. 016)in 70 bpm≤RHR ＜80 bpm group and 1.640(95% CI:1.067 -2.523,P =0. 024)in 80 bpm ≤ RHR group,and the OR values for hypercholesterolemia was 1.197(95 % CI: 0.749 - 1.913,P=0.453)in70 bpm≤RHR ＜80 bpm group and 1.814(95% CI:1. 036-3. 177,P =0.037)in 80 bpm≤RHR group. Conclusions RHR is independently and positively correlated with the prevalence of hypertriglyceridemia and hypercholesterolemia and the blood levels of both lipid. RHR has no significant correlation with HDL-C or LDL-C level.

Objective:To study MUC1 based cancer vaccine.Methods:MUC1 gene was inserted into pMAL-p2 vector and constructed recombinant pMAL-MUC1. MUC1-MBP fusion protein expression was induced by IPTG in E coli DH5? transformed by the recombinant pMAL-MUC1 .The fusion protein was analyzed by Western blot and purified by amylose affinity chromatography. The antiserum,T cell proliferation and CTL activity of spleen from C57 mice immunized by MUC1-MBP were determined respectively by ELISA,adding 3H-TdR and MTT.Results:Had successfully constructed pMAL-MUC1 expression vector,and purified MUC1-MBP and MBP. C57 mice immunized by MUC-MBP generated MUC1 specific antibody and CTL.The titer of polyclonal antibody to MUC1 was about 1∶5 760?3 221. CTL cytotoxicity to the MCF7 and lewis lung cancer cells respectively were at 47.7%?4.3% and 67.5%?6.5%.Conclusion:Human recombinant MUC1-MBP fusion protein activated T and B cell response in mice.The results suggested that the recombinant Muc1 may be used to develop protein vaccine against carcinoma.

Objective To investigate the alteration of protein kinase C (PKC) and endothelin system in early diabetic rats, and the effect of specific PKC inhibitor on the expression of retinal endothelin-1 (ET-1). Methods The rats model with streptozotocin(STZ)-induced diabetes were set up. The expression of retinal PKC was detected by enzyme-linked immunoabsorbent assay (ELISA). The expression of retinal ET-1, ET-3, ET-A and ET-B receptor mRNA was determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The alteration of retinal ET-1 mRNA after intravitreal injection of PKC inhibitor GF109203X in diabetic rats was also observed. Results The activities of membranous PKC were significantly increased in 2-week diabetic rats compared with that in normal rats(t=3.296, P=0 008), while activities of cytosolic PKC were unchangeable(t=0 138, P=0 894). The expression of retinal ET-1 mRNA was significantly increased(P=0 008), while no change was found in expression of ET-3, ET-A and ET-B mRNA(P=0 918,P=0 889,P=0 500). After intravitreal injection of 10 -5、10 -6、10 -7 mol/L PKC inhibitor GF109203X in diabetic rats, the expression of retinal ET-1 mRNA was decreased in a dose-dependent manner compared with the control rats. Conclusion Activation of PKC and increased expression of ET-1 could be found in the retina of early diabetic rats, and PKC inhibitor could inhibit the expression of retinal ET-1.

Objective To investigate endoscopic clipping in treating acute gastroduodenal perforation. Methods Endoscopic clipping was carried out in 22 patients with acute gastroduodenal perforation as the complication of peptic ulcer or endoscopy. Results The clipping was successful in 19(86. 4% ) of 22 patients with acute gastro duodenal perforation, and surgical therapy was performed in 3(13. 6% ) of 22 cases. A single clipping was performed in 14(73. 7% ) of 19 patients resulted in satisfactory closure of perforation. An additional clipping was needed in 5(26. 3% ) patients 24 h after the first endoscopes procedure. The effectiveness of endoscopic clipping was related to location, cause and size of the perforation. Conclusion Endoscopic clipping is an effective method for acute gastroduodenal perforation as the complication of peptic ulcer or endoscopy.