Determination of squamous cell carcinoma antigen based on the magnetic particles chemiluminescence immunoassay / 国际检验医学杂志

Objective To establish a rapid and sensitive chemiluminescence immunoassay for detecting squamous cell carcinoma antigen (SCCA)in human serum.Methods Fluorescein isothiocyanate (FITC)and N-(aminobutyl)-N-(ethylisoluminol)(ABEI) were used to label two different monoclonal antibodies of anti-SCCA.SCCA in serum combined with labeled antibodies and formed a sandwiched immunoreaction.After adding the substrate solution,the relative light unit of ABEI was measured.Magnetic particles coated with anti-FITC antibody were used as solid separation carrier.Results The results demonstrated that the method was linear to 22 ng/mL with a detection limit of 0.025 ng/mL.The coefficient of variation (CV)was less than 6% and 7% for intra-assay and inter-assay precision,respectively.Compared with the commercial kit,the proposed method showed a correlation of 0.990 1.Conclu-sion chemiluminescence immunoassay based on magnetic particles displays acceptable performance for quantification of SCCA and is appropriate for use in clinical diagnosis.

Expression characteristics of 123I-vascular endothelial growth factor-binding sites on tumor cells / 国际肿瘤学杂志

Objective To explore the expression characteristics of vascular endothelial growth factor (VEGF) receptor (VEGFR).Methods The 123I-VEGF165 and 123I-VEGF121 were marked to human umbilical vein endothelial cell (HUVEC),several human tumor cell lines (HMC-1,A431,KU812,U937,HEP-1,HEP-G2,HEP-3B and Raji),a variety of human tumors and adjacent non-neoplastic tissues as well as peripheral blood cells.Then,the specific binding site maximal binding capacity (Bmax),dissociation constant (Kd) and concentration of 50％ required specific binding (IC5o) were analyzed.The affinity,quantity and specificity of different cells combined with 123I-VEGF165 and 123I-VEGF121 were judged.Results Two kinds of analogous 123I-VEGF165 binding sites on the surface of HUVEC were found.While,there was only one kind of 123I-VEGF121binding site.123I-VEGF121 was found on the special cell lines (HUVEC,HEP-1 and HMC-1) and special early tumors (early melanoma,ductal breast cancer,ovarian cancer and meningioma).Compared with peripheral blood cells and adjacent non-neoplastic tissues,the number of VEGFR of tumor cells was bigger.Among the 123I-VEGF165 marked VEGFR,the Bmax value of early melanoma,ductal breast cancer,hepatocellular carcinoma,papillary thyroid carcinoma,ovarian carcinoma,renal cell carcinoma were 45 ± 13,13 ± 3,25 ±8,5 ±2,42 ± 12,20 ±6,respectively.While among the 123I-VEGF121 marked VEGFR,the Bmax value of early melanoma,ductal breast cancer,ovarian carcinoma were 30 ± 8,8 ± 3,20 ± 6.123I-VEGF165 and 123I-VEGF121 had specific binding capacity with a variety of human tumor cells and tissues.Compared with 123I-VEGF121,there were more different kinds of tumor cells could be bound to 123I-VEGF165 with higher capacity.Conclusion 123I-VEGF165 may be a potential target of tumor imaging in vivo,and it is expected to be used to diagnose and treat tumors.

CpG Array Analysis of Histone H3 Lysine 4 Trimethylation by Chromatin Immunoprecipitation Linked to Microarrays Analysis in Peripheral Blood Mononuclear Cells of IgA Nephropathy Patients

PURPOSE: The purpose of the present study was to investigate the aberrance of histone H3 lysine 4 trimethylation (H3K4me3) in patients with IgA Nephropathy (IgAN). MATERIALS AND METHODS: In this study, H3K4me3 variations in peripheral blood mononuclear cells (PBMCs) from 15 IgAN patients and 15 healthy subjects were analyzed using chromatin immunoprecipitation linked to microarrays analysis (ChIP-chip). ChIP real-time PCR was used to validate the microarray results. Expression analysis by quantitative real-time PCR (qRT-PCR) revealed correlations between mRNA and H3K4me3 levels. DNA methylation status was analyzed by quantitative methylation-specific PCR. RESULTS: We found that 321 probes displayed significant H3K4me3 differences in IgAN patients compared with healthy controls. Among these probes, 154 probes displayed increased H3K4me3 and 167 probes demonstrated decreased H3K4me3. For further validation, we selected 4 key relevant genes (FCRL4, GALK2, PTPRN2 and IL1RAPL1) to study. The results of ChIP real-time PCR coincided well with the microarray data. Quantitative RT-PCR revealed the correlations between the mRNA expression and the methylation levels of H3K4me3. Different degrees of DNA methylation alterations appeared on the selected positive genes. CONCLUSION: Our studies indicated that there were significant alterations in H3K4me3 in IgAN patients. These findings may help to explain the disturbed immunity and abnormal glycosylation involved in IgAN patients.

Research progress on induction of pluripotent stem cell from adult stem cell / 国际生物医学工程杂志

It is confirmed that the adult cells can be re-programmed to embryonic stem cells(ESCs) by presenting some certain factors in oocytes in the clone process of animals. In recent years, some transcription factors that can induce pluripotent stem cells(iPS) have been identified and which made it possible to obtain induced pluripotent stem cells similar to embryonic stem cells. iPS provides a unique platform to study the pluripotent mechanism and to establish some specific disease models. This major scientific discovery can not only avoid the use of ES which involves ethics debate, but also lead the stem cell research to a new field.

Application of iTRAQ multiple labeling and tandem mass spectrometry in proteomic research of human peripheral blood mononuclear cells / 生物医学工程学杂志

This paper is aimed to establish and optimize proteomic research platform using isobaric tags for relative and absolute quantitation (iTRAQ) so as to facilitate further proteomic research of human peripheral blood mononuclear cells (hPBMC). We collected hPBMC, after protein extraction and trypsin digestion, we labeled the samples with iTRAQ reagents and then subjected to mass spectrometry. In triplicates, thirty precursors were randomly selected and detected; as a result, 26, 28 and 29 peptides were respectively tagged with ITRAQ reporter ions. The labeling efficiencies ranged between 86.7%-96.7%, with no significant difference among the groups (P>0.05). The coefficient of variance for the relative ratios of peptides from different proteins was ranged from 7.6% to 25.5% and there were no significant differences across the groups (P>0.05). The coefficient of variance for the relative ratios of different peptides from the same protein was varied from 9.3% to 19.1% and the differences across groups were not significant (P>0.05). The labeling of iTRAQ combined with tandem mass spectrometry in PBMC was successful with favourable reproducibility and accuracy, which could lay a foundation for further proteomic study of hPBMC in autoimmune disorders.

MtrF gene expression in Neisseria gonorrhoeae of different resistance levels / 西安交通大学学报(医学版)

Objective To study the relationship between the expression of mtrF gene and high-level multiple resistant Neisseria gonorrhoea.Methods ① The susceptibility of 58 clinical Neisseria gonorrhoeae to 5 kinds of antibiotic agents was tested by disc diffusion method.② The minimum inhibitory concentration(MIC) of erythromycin was determined by tube dilution method.③ The expression of mtrD and mtrF gene in susceptive group,inter-mediated resistant group and high-level multiple resistant group was detected by semi-quantitative RT-PCR.Results ① There were 30 strains presenting resistance to two or more than two antimicrobial agents,which accounted for 51.7% of the 58 clinical strains.② The number of strains sensitive,intermediate and resistant to erythromycin was 7,21 and 30,respectively,and there were 17 strains with erythromycin MIC≥32.0 ?g /mL.③ Compared with that in susceptive group and inter-mediated resistant group,mtrF expression was up-regulated in high-level multiple resistant group(P