Value of Serum Free Light Chain Kappa/Lambda Ratio Detection Combined with Immunofixation Electrophoresis in the Prognosis Evaluation of Patients with Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To analyzed the prognostic value of serum free light chain kappa/lambda ratio detection combined with immunofixation electrophoresis in multiple myeloma (MM) patients.@*METHODS@#72 patients with MM treated in our hospital from January 2017 to December 2018 were selected. Serum free light chain kappa/lambda ratio (sFLCR) and immune typing were detected respectively. The clinical characteristics and survival time were compared among patients. COX regression was used to analyze the factors influencing prognosis.@*RESULTS@#38 patients showed high sFLCR, and 34 showed low sFLCR. Compared with the low sFLCR group, the DS stage of patients in high sFLCR group elevated, the levels of β2-MG and Scrwere increased, and Hb decreased, all the differences were statistically significant (P＜0.05). Among 72 patients, there were 40 cases of IgG type (55.56%), 27 cases of IgA type (37.50%) and 5 cases of IgM type (6.94%). Compared with IgG and IgA patients, the serum calcium and creatinine in IgM patients were increased significantly, while Hb decreased significantly (P＜0.05). The median survival time was 19.2 months in 21 patients with IgG type and high sFLCR; 24.0 months in 19 patients with IgG type and low sFLCR; 15.0 months in 12 patients with IgA type and high sFLCR; 16.7 months in 15 patients with IgA type and low sFLCR; 6.0 months in 5 patients with IgM type and high sFLCR,respectively. DS stage, M protein typing and sFLCR correlated with prognosis of patients (P＜0.05).@*CONCLUSION@#The serum free light chain kappa/lambda ratio combined with immunofixation electrophoresis is valuable for the prognostic evaluation of patients with multiple myeloma.

Prokaryotic expression of DNA recombinase FLP and its purification with enzymatic activity / 生物工程学报

DNA recombinase FLP gene exists on the 2 micro plasmid of Saccharomyces cerevisiae. Recombinase FLP could recognize an FRT site composed of 34bp and function the sequences for exchange, recombination, deletion and reversion between the two orientated FRT sites. These functions are highly recognized by molecular biologists and biotechnology engineers for theoretic and applicable technology studies. This work constructed a prokaryotic over-expressed vector harboring FLP gene nominated as pQE30-flpe and established its over-expression culture system in which recombinase FLP could be efficiently expressed in E. coli strain M15. Purification procedures for high purity and active FLP are established through combination of ammonium sulfate precipitation with a 0.5-1.0 mL micro-column technique of Ni affinity chromatography with gradient elution. To verify the recombinase activity of purified FLP, substrate vectors, sequence donor vector (pUC18-FRT-gfp-FRT) and sequence accepting vector (pET30a-FRT) are constructed with various number, orientation of FRTs harboring the GFP gene for the expression of visible assay of the functions of recombination, exchange and deletion. Results showed that the system not only over expressed recombinase FLP in prokaryotic E. coli, but also efficiently purified the enzyme with a higher activity of the function of recombination, exchange and deletion. The system and the method are easily implemented and feasibly manipulated for theoretic study and biotechnology application.

Expression and clinical significance of HBsAg and HBcAg in hepatocytes in chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the expression of HBsAg and HBcAg in hepatocytes in CHB patients, and analyze the correlation among the expression of HBsAg and HBcAg, the quantity of HBV DNA in serum, the pathology of liver tissue and the clinical manifestation.</p><p><b>METHODS</b>Quantitative polymerase chain reaction was used to assay the quantity of HBV DNA in serum in 351 CHB patients. Furthermore pathological diagnosis was performed using liver biopsy to assay the expression of HBsAg and HBcAg in hepatocytes by an immunohistochemical staining technique.</p><p><b>RESULTS</b>The positive expression rate of HBsAg and HBcAg in hepatocytes was 92.3% and 76.9% respectively. Cytoplasm-membrane HBcAg expression type (75.6%) was observed in the CHB with more active inflammation, while Nucleus HBcAg expression type (24.4%) was observed in the CHB with more sedative one (P < 0.0001). The expression of HBsAg was correlated with the quantity of HBV DNA in serum (rp = 0.24, P = 0.0129), while inversely correlated with the inflammation and the fibrillation of liver tissue (rp = -0.22, P = 0.0279; rp = -0.23, P = 0.0186). The expression of HBcAg was correlated with the quantity of HBV DNA in serum (rp = 0.52, P < 0.0001), while was inversely correlated with the inflammation and the fibrosis of liver (rp = -0.33, P < 0.0001; rp = -0.34, P < 0.0001).</p><p><b>CONCLUSION</b>Cytoplasm-membrane HBcAg expression type was observed in the CHB with more active inflammation, while Nucleus HBcAg expression type was observed in the CHB with mild change. In the immunopathogenesis of the liver damage in CHB, HBcAg might be a main target antigen. HBsAg might be a sensitive index to screen HBV infection; HBcAg might probably be a reliable index to evaluate the replication of HBV</p>