Identification of the IgE-binding epitopes in main dust mite allergen Der p 1 / 南方医科大学学报

<p><b>OBJECTIVE</b>To identify the IgE-binding epitopes in the allergen Der p 1 of main house dust mites, which can be recognized by the specific IgE in the sera from allergic individuals, and obtain a hypoallergen derived from the T-B epitope fused peptide for potential use in specific immunotherapy (SIT).</p><p><b>METHODS</b>Thirty-one peptides containing 15 amino acids each, which covered the full 222 amino acids of Der p 1 protein sequence, were synthesized on the cellulous membrane by solid-phase peptide (SPOTs) synthesis, with 8 overlapping amino acids between every two neighboring peptides. The membrane bearing the spots of the synthesized peptides were incubated with the allergic serum pools consisting of the sera from 5 allergic individuals. The membrane was then probed with HRP-conjugated anti-human IgE, followed by enhanced chemiluminescence (ECL) for visualization and gray scale analysis of the positive peptide spots.</p><p><b>RESULTS</b>Three strong IgE-binding epitopes were identified in the amino acid sequence of Der p 1 molecule, namely Ep1 (amino acids 85-99), Ep2 (amino acids 106-120) and Ep3 (amino acids 190-204).</p><p><b>CONCLUSION</b>The 3 IgE-binding epitopes (B cell epitopes) identified in Der p 1 confirm the presence of linear epitopes in Der p 1, suggesting the possibility of constructing T/B epitope-fused hypoallergens.</p>

Expression, purification and identification of Abeta fusion protein with enhanced solubility / 南方医科大学学报

<p><b>OBJECTIVE</b>To economically obtain the Abeta peptide for Alzheimer's disease (AD) research by expressing the Abeta peptide fused with the maltose binding protein (MBP) possessing high solubility in E.coli.</p><p><b>METHODS</b>The cDNA-coding sequence of Abeta peptide was modified by the addition of a BamH I site at the 5' end and a Hind III site at the 3' end using PCR. The modified sequence was ligated into the maltose-binding protein (MBP) fusion expression vector pMAL-c2 containing an thrombin cleavage site, which was transformed into competent E.coli DH5alpha cells. After identification of the single clones by PCR and DNA sequencing, the recombinant plasmid was transformed into E.coli TB1 and induced to express MBP-Abeta fusion protein. The expressed fusion protein was purified using amylose resin column and identified by SDS-PAGE and Western blotting.</p><p><b>RESULTS</b>The result of DNA sequencing verified the consistency between the inserted sequence and Abeta (1-42) sequence. SDS-PAGE electrophoresis showed that MBP-Abeta fusion protein was highly expressed in E.coli TB1, and Western blotting demonstrated that the purified fusion protein and the separated Abeta peptide could be recognized by specific anti-Abeta (22-35) antibody.</p><p><b>CONCLUSION</b>MBP-Abeta fusion protein highly expressed in E. coli TB1 cells with enhanced solubility and the separated Abeta peptide with good immunogenicity obtained may lend support to AD research.</p>

Cardioprotection of Shenfu Injection against myocardial ischemia/reperfusion injury in open heart surgery / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of Shenfu Injection (SFI) against myocardium ischemia/reperfusion injury (IRI) in mitral valve replacement (MVR) with cardiopulmonary bypass (CPB).</p><p><b>METHODS</b>Forty patients undergoing selective MVR were randomly assigned to the control group and trial Groups I, II, III, and IV according to the different administrations of SFI, 8 patients in each group. The changes of systolic blood pressure (SBP), mean blood pressure (MBP), diastolic blood pressure (DBP) in each group were monitored, respectively. The recovering percentage of spontaneous heart beat, the heart rate (HR) and cardiac rhythm as well as the abnormal duration of ECG-ST segment were recorded after the restoration of heart beat. The serum concentration of cardiac troponin I (cTnI), malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were determined as well.</p><p><b>RESULTS</b>(1) The SBP, MBP and DBP values, the recovering rate of spontaneous heart beat, HR, ECG-ST, atrioventricular block and ventricular arrhythmia were significantly improved in group IV compared with any other groups. (2) Compared with the control group, the postoperative serum contents of cTnI and MDA were significantly decreased, but the activity of SOD was significantly increased in group IV.</p><p><b>CONCLUSIONS</b>SFI had a certain protective effect against myocardium IRI. Moreover, better efficacy was seen with the administration of 1.5 mL/kg SFI into CPB priming fluid and pumping 1.5 mL/kg SFI via CPB as soon as the clamped aorta was unclamped.</p>

Exploration on the effect and mechanism of shenfu injection on resuscitation from general anesthesia / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect and the mechanism of Shenfu injection (SFI) on the resuscitation from general anesthesia.</p><p><b>METHODS</b>Forty patients who received selective abdominal surgery with general anesthesia for 3-4 hrs and ASA grade I-II were divided into two groups, the trial group and the control group, 20 patients in each group. After being sent into the postanesthesia care unit (PACU), the trial group was treated with intravenous dripping of SFI 1.0 ml/kg and the control group was treated with intravenous dripping of equal volume of normal saline. All patients were observed in double blindly manner, the self ventilation recovery time, extubation time, the time of leaving PACU and their Glasgow coma scale (GCS) were recorded and compared. 2 ml of peripheral venous blood were taken to determine the plasma beta-endorphin (beta-EP) content at the time points of before (T1), 5 min (T2), 15 min (T3) and 30 min (T4) after dripping.</p><p><b>RESULTS</b>The self ventilation recovery time, extubation time and time of leaving PACU in the trial group were all shorter than those in the control group (P < 0.01), the GCS in the trial group was better than that in the control group (P < 0.01). The plasma content of beta-EP raised gradually along the recovering of patients consciousness, as compared with the content before dripping (T1), it showed insignificant difference at time point T2 but significant difference at T3 and T4 comparison at the corresponding time point showed that the content at T1 and T2 were similar in the two groups (P > 0.05), but at T3 and T4, the content was higher in the trial group than that in the control group respectively (P < 0.01).</p><p><b>CONCLUSION</b>SFI could accelerate the resuscitation after general anesthesia, the mechanism may be related with its action in raising plasma beta-EP level.</p>