ABSTRACT
OBJECTIVE: To investigate the value of serum galactose-deficient IgA1 in the diagnosis of IgA nephropathy and explore its relationship with the decline of renal function and pathological grade of renal biospy samples of the patients. METHODS: The serum samples were collected from, Shengjing Hospital of China Medical Unerversity from January 2016 to December 2017,which included 40 IgA nephropathy patients(group A), 20 other primary glomerulonephritis patients(group B) and 20 healthy persons(group C).Serum levels of GD-IgA1 were detected in all the samples.The 24-hour urinary protein and serum creatinine were measured in group A and B,the eGFR calculated. Recorded the pathological grade of renal biospy samples and Lee's classification. Study the value of serum Gd-IgA1 level in diagnosing IgA nephropathy by drawing ROC curve and calculating the area under the curve. RESULTS: The serum levels of GdIgA1 in IgA nephropathy patients were significantly higher than those in other types of primary glomerular diseases and healthy controls.The area under ROC curve was 0.886.When serum Gd-IgA1 level is higher than 662.5 U/ml, it suggested that people were more likely to have a IgA nephropathy.The level of serum Gd-IgA1 was related to the decline of renal function and pathological grade of renal biospy samples in patients with IgA nephropathy. CONCLUSION: Serum Gd-IgA1 levels may be helpful in the diagnosis of IgA nephropathy in patients who can not undergo renal pathological examination.
ABSTRACT
Electrospray ionization is most commonly used in mass spectrometry for analysis of biological molecules such as peptides and proteins. However, peptides and proteins ions produced by electrospray ionization usually have multiple charges, and produce multiple spectrum peaks, making the mass spectrum complicated. Gas phase proton transfer ion/ion reaction can effectively regulate the charge states of peptides and proteins ions after electrospray ionization, simplifying the spectrogram, and thus is significant to the analysis of complex proteins and peptides samples. In this study, a dual-polarity linear ion trap ( LIT) mass spectrometer was used to control the charge state of peptide ions by proton transfer ion/ion reaction. The detection effect of the method was verified by using glutathione ( oxidation type) , oxytocin and dynorphin as typical samples. The results showed that this method could remove excess charges in the positive ions. When injecting enough negative ions for eaction, peptide ions with multiple charge valence state could be reduced to the minimum, therefore simplify the spectrogram effectively.
ABSTRACT
A pair of degenerate primers were designed based on NBS (nucleotide binding site, NBS) domain of resistance(R) gene and used to perform PCR with cDNA from the translocation line 6VS/6AL of Triticum aestivum-Haynaldia villosa. A clone (N7) characterized with NBS was obtained by sequencing analysis. Two specific primers were designed from the N7 sequence and used to screen a genomic TAC (transformation-competent artificial chromosome, TAC) library of 6VS/6AL consisting of ca. 2 x 10(6) clones. The library was stored as clone pools in twenty-two 96-well plates, each well containing approximately 1000 TAC clones. TAC plasmids were prepared from all the 2112 pools. Using a pooled PCR screening procedure, a positive TAC clone having a 40 kb insert was obtained. The positive clone was confirmed by Southern hybridization with the NBS fragment as a probe. The results indicate that the pooled PCR method is effective for screening of genomic libraries having large number of clones.