Development and application of CK-MB specific monoclonal antibodies / 生物工程学报

The aim of this study is to develop creatine kinase isoenzyme MB (CK-MB) specific monoclonal antibodies (mAb), and characterize the monoclonal antibody and further development of quantitative detection assay for CK-MB. The BALB/c mice were immunized with purchased CK-MB antigen, then monoclonal antibodies were prepared according to conventional hybridoma technique and screened by indirect and capture ELISA method. To identify the epitopes and evaluate the classification, purchased creatine kinase isoenzyme MB (CK-MM/BB/MB) antigen was used to identify the epitopes, with immunoblotting and synthetic CK-MM and CK-BB in different linear epitope. A double antibody sandwich ELISA was applied to screen the mAb pairs for CK-MB detection, and the quantitative detection assay for CK-MB was developed. We used 74 cases of clinical specimens for comparison of our assay with Roche's CK-MB assay. We successfully developed 22 strains of hybridoms against CK-MB, these mAbs can be divided into linear, partial conformational CK-MB, CK-MM or CK-BB cross monoclonal antibody and CK-MB specific reaction with partial conformational monoclonal antibody, and CK-MB quantitative detection assay was developed by using partial conformational monoclonal antibody. The correlation coefficient factor r of our reagent and Roche's was 0.930 9. This study established a screening method for CK-MB partial conformational specific monoclonal antibody, and these monoclonal antibodies were analyzed and an established quantitative detection assay was developed. The new assay had a high concordance with Roche's.

Development, identification and application of 33 monoclonal antibodies against cardiac troponin T / 生物工程学报

The aim of this study is to prepare and characterize cardiac troponin T (cTnT) monoclonal antibodies (mAb), and further develop a chemiluminescence quantitative detection assay for cTnT. BALB/c mice were immunized with recombinant cTnT antigen, and specific mAbs were prepared using conventional hybridoma technique and screened by indirect ELISA method. To identify the epitopes, several cTnT peptide fragments were synthesized or expressed by genetic engineering. A double antibody sandwich ELISA method was used to screen the mAb pairs for cTnT detection, and the automatic chemiluminescence detection assay for cTnT was developed. In total 220 clinical specimens were used for system comparison between our assay and Roche cTnT assay; further performance characteristics was evaluated by testing 238 clinical samples and 784 physical examination samples. We successfully screened 33 strains of hybridoms against cTnT, and the mAbs' epitopes were identified. Mab E16H8 and C8G11 with a detection limit of 10 pg/mL cTnT antigen were selected to develop the full automatic chemiluminescence quantitative assay. The correlation coefficient of our reagent with Roche's was 0.959 9, with a coincidence rate of 95%. The assay presented a sensitivity of 97.5%, and a specificity of 99.15% in detection of clinical samples. The cTnT concentration was less than 0.080 6 ng/mL in 99% of general population, which agrees with the definition of WHO on patients with acute myocardial infarction (AMI). In summary, we developed monoclonal antibodies against predominant epitopes for diagnostics of cTnT, and an automatic tubular chemiluminescence quantitative detection assay was further developed, which presents a high coincidence rate with Roche's.

Pulchinenoside inhibits the fibroblast-like synoviocytes apoptosis in adjuvant arthritis rats / 中南大学学报(医学版)

OBJECTIVE@#To explore the eff ect of pulchinenoside (PULC) on fi broblast-like synoviocytes (FLS) apoptosis in adjuvant arthritis (AA) rats.@*METHODS@#A total of 60 SD rats were randomly divided into 8 groups: A normal control group, an AA group, a low PULC group (50 mg/kg), a middle PULC group (100 mg/kg) or a high PULC group (150 mg/kg) and an ibuprofen (8 mg/kg) group (n=10 per group). FLS from the AA rats was cultured. The expression of Bcl-2, Bax, caspase-3 and the FLS proliferation were detected by the real time qPCR and MTT, respectively. The expression of IL-6 and IL-8 in culture medium was detected by ELISA.@*RESULTS@#Compared with the AA group, the Bcl-2 expression was down-regulated (all P<0.05), the Bax and caspase-3 expression was up-regulated (all P<0.05), and the FLS proliferation was inhibited (all P<0.05). The IL-6 and IL-8 expression was suppressed in the FLS in the PULC groups at different dosages (all P<0.05) as well as in the ibuprofen group (P<0.05).@*CONCLUSION@#PULC may inhibit the FLS proliferation in AA rats by increase in FLS apoptosis.

Effect of taper and inserting angle on stability of micro-implants in type II bone with three-dimensional finite element / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the effect of taper and inserting angle on stability of micro-implants in type II bone and to provide theoretical evidence for clinical selection of taper and inserting angle.</p><p><b>METHODS</b>Three-dimensional finite element models of type II bone sections and micro-implants were fabricated. They were designed using different tapers (diameter between 1.1 to 1.6 mm, taper between 0 to 0.0625) and different inserting angles ranging from 30 degrees to 90 degrees (30 degrees, 45 degrees, 60 degrees, 75 degrees, 90 degrees). Effects on the maximum von Mises stresses in mandible and micro-implants, as well as on the maximum displacements in micro-implants, were analyzed under an applied 2 N horizontal force.</p><p><b>RESULTS</b>The inserting angle had a significant effect on stresses and displacements, with lower distribution of stresses and displacements occurring at 60 degrees. The effect of the taper was also related to the inserting angle. The minimum stresses were 5.013 4 MPa (cortical bone) and 25.1310 MPa (micro-implant) in model D at 60 degrees. The displacements of micro-implants were proportional to the taper.</p><p><b>CONCLUSION</b>The inserting angle has a significant effect on stability of micro-implants, and the recommended inserting angle is 60 degrees. The influence of taper on the stability of micro-implants is related to the inserting angle. The micro-implant with 0.0375 taper (maximum diameter is 1.6 mm, minimum diameter is 1.3 mm) is more suitable for type II bones.</p>

Approach to the way of classroom teaching on electrocardiogram / 中华医学教育探索杂志

That there are so many difficulties in both of teaching and learning electrocar-diogram(ECG)is the characteristics in teaching and studying ECG. The following ways are the ef-fective methods of improving effects of classroom teaching of ECG:emphasizing the importance of ECG in clinical work at the beginning of the lecture on ECG in order that the students can understand the necessity of studying ECG and make up their mind to study ECG well;studying the teaching contents on ECG seriously and supplement the textbook appropriately;set up the link of discussion and create the condition that teacher and students work altogether;produce the slides with pictures(mainly)and words and give accounts of the pictures mainly;sum up the character-istics of all sorts of abnormal ECG and state the characteristics of presenting ECG simultaneously.