Objective To investigate the clinical value of immunophenotyping and quantitative analysis of serum immunoglobulin in the diagnosis and typing of monoclonal gammaglobulinemia.Methods 118 serum samples were collected from the patients with monoclonal gammaglobulinemia and performed the serum protein electrophoresis(SPE),serum immunofixation(IF)electrophoresis, immunoglobulins(Ig)quantitation and serum light chain κ,λdetection.The analysis was conducted aiming at the patients with im-munophenotyping positive and the Ig quantitation negative.Results 56 cases showed immunophenotyping positive and the immuno-globulin quantitation negative,among them,6 cases were free light chain type (κlight chain in 2 cases andλlight chain in 4 cases), 1 case was the non-secretion type;62 cases showed immunophenotyping positive and the Ig quantitation positive.Conclusion The IF technique has an important significance for the diagnosis and typing of monoclonal gammaglobulinemia,its diagnosis can be com-bined with the other laboratory tests of serum Ig and light chain quantitative detection.

Flow cytometry (FCM) is one of the most advanced cell quantitative analysis technology.Today,flow cytometry has been extensively and intensively used in medicine and other science,ranging from basic research to clinical diagnosis With the rapid development of science and technology of China,the application of flow cytometry in clinical research and diagnosis has also made significant progress.

Objective To investigate BRAFV600E mutation and sodium iodide symporter (NIS) expression and its clinical significance in papillary thyroid carcinomas (PTC).Methods BRAFV600E mutation was evaluated by direct sequencing and BRAFV600E mutation was determined in 40 cases of PTC.NIS expression was examined by means of immunohistochemistry.The relationship between BRAFV600E mutation and the clinicopathologic features of PTC was analyzed.The relationship between NIS expression and BRAFV600E mutation in PTC was analyzed.Results Positive BRAFV600E mutation was determined in 23 of 40 cases of PTC (57.5％),whereas positive NIS expression was showed in 7 of 40 cases (12.5％).BRAFV600E mutation was associated with extrathyroid invasion and a high risk of disease recurrence in PTC (P ＜ 0.05).Positive NIS expression was found in only one of 23 cases of PTC with BRAFV600E mutation,and positive NIS expression was associated with a statistically significant lower BRAFV600E mutation in PTC (P =0.011).Conclusions BRAFV600E mutation might be associated with higher aggressiveness,higher disease recurrence and a poorer prognosis.Lower NIS expression may be responsible for poor 131I uptake in

The expression of Rap1 GAP protein was detected in 69 cases of papillary thyroid carcinoma and adjacent normal thyroid tissues with immunohistochemistry method.Methylation of Rapl GAP gene was analyzed by methylation-specific-PCR (MSP) in these tissues.The immunohistochemistry results indicated that the Rap1GAP protein was down-regulated in tumor tissues of 54 ( 78％ ) cases compared with normal thyroid tissues.Statistical analysis demonstrated that the decreased level of Rap1 GAP protein expression was significantly correlated with the tumor stage T according to American Joint Committee on Cancer ( AJCC,P =0.043 ).The MSP results demonstrated that 46 cases of Rap1GAP gene methylation were detected in 54 cases with down-regulated expression of Rap1GAP (66.7％),but only 1 case of methylation was found in 15 cases without obvious change of Rap1GAP expression (6.67％),showing significant difference ( P＜0.01 ).There was no methylation in normal thyroid tissues.These results suggest that raised methylation of promoter region may contribute to the low expression of Rap1GAP protein in papillary thyroid carcinoma.

Object To study the isolation and purification of a polysaccharide, obtained from Paecilomyces tenuipes Samson, its molecular weight, sugar composition, and mode of linkage Methods Crude polysaccharide was extracted by water at ambient temperature and purified on Sephadex G 100 column Its monosaccharide composition was determined by ionic ion exchange column after complete hydrolysis with acid Their mode of linkage was determined by methylation and glycosidic linkage established by IR and NMR spectra Results HPLC spectrum showed that the polysaccharide was of homogeneous composition, which was also proved latter by GC MS and NMR Conclusion Polysaccharide obtained from P tenuipes Samson is ? (1→6) linked and composed of only D glucose The molecular weight was 2 05?10 4

opping drinking. They were not influenced by gender, smoking and drinking histories. They could serve as monitoring indexes for recent drinking status on healthy individuals.

Objective To explore the levels of serum chemokine in the patients with gastric cancer using cytometric beads array (CBA) and to find out the laboratory evidence for gastric cancer immunotberapy.Methods Forty-five patients with gastric cancer,thirty patients with benign gastric diseases and forty healthy controls were included.The chemokine levels of interleukin-8 (IL-8),regulated upon activation normal T-cell expressed and secreted (RANTES),monokine induced by IFN-γ (MIG),monocyte ehemoattractant protein-1(MCP-1) and interferon-γ induced protein-10 (IP-10) in serum of these three groups were measured using CBA ,then these data were analyzed using BD CBA analysis software.Results The levels of serum chemokines in healthy controls were (176.4±20.7) μg/L for IP-10,(111.3±17.2) μg/L for MCP-1,(503.9±47.2) μg/L for MIG,(472.4±116.7) μg/L for IL-8,respectively.In gastric cancer patients,the levels were (266.6±24.7) μg/L for IP-10,(100.4,70.8-193.5) μg/L for MCP-1,(1614±275.4) μg/L for MIG,(500.0±164.8) μg/L for IL-8.The levels of serum chemokines in patients with benign gastric disease were (207.9±31.7) μg/L for IP-10,(121.2±23.6) μg/L for MCP-1,(514.5±63.0) μg/L for MIG,(480.2±134.8) μg/L for IL-8,respectively.The levels of IP-10 and MIG among these three groups were significantly different (IP-10:F = 3.52,P < 0.05,MIG:F = 9.27,P < 0.01).The levels of IP-10 were elevated in the cancer patients compared with healthy controls (P < 0.05),while the levels of MIG were elevated significantly in the cancer patients compared with healthy controls and benign disease controls(t=3.29,P < 0.01,t=2.84,P<0.01).Conclusions CBA is a novel method with convenience,sensitivity and accuracy for the detection of serum ehemokines.The concentration of IP-10 and MIG in gastric cancer patients are elevated,indicating its implication in cancer pathogenesis and metastasis.Measurement of serum chemokines will lay the groundwork for therapeutic strategy in gastric cancer by inhibiting and anti-chemokine.

Objective To investigate the expression of galectin-10 in CD4+CD25+CD127low/-regulatory T cells(Treg) and the correlation between expressions of galectin-10 and Foxp3.Methods CD4+CD25+CD127low/-Treg cells and CD4+CD25-CD127+ T cells were isolated from PBMC of healthy volunteers and lymph nodes of gastric cancer patients by flow cytometry.Following RNA micro-extraction RT-PCR analysis was used to detect mRNA expression of galectin-10 and Foxp3.Results The mRNA expression of galectin-10 and Foxp3 were dominant in Treg cells,whereas nearly no expression in CD4+CD25-CD127+ T cells.The results from PBMC of healthy volunteers and lymph nodes of gastric cancer patients were consistent between.Conclusion The expression of galectin-10 was obviously high in Treg cells,so it may become a novel marker for phenotypic characterization of Treg cells.

Objective To evaluate the consistency and accuracy among 3 brands of flow cytometers (BriCyte E6,BD FACSCanto Ⅱ and Beckman Coulter FC 500) in the detection of lymphocyte subsets.Methods According to the methodology,the BriCyte E6 was compared with 2 flow cytometers commonly used in clinical detection.Seventy-three cases (40 male and 33 female) of anticoagulation peripheral blood specimens were collected in the clinical laborartory department of Xinhua Hospital in July 2015 and the percentage (％) and absolute number (#) of the lymphocyte subsets were detected by 3 different flow cytometers within samples collected 4 h.Results There were good consistency among the 3 flow cytometers (R2 ＞0.95,R2 from 0.969 5 to 0.992 4) in the detection of lymphocyte subsets percentage,so did in the detection of absolute number (R2 ＞ 0.95,R2 from 0.969 1 to 0.993 3).As to the precision evaluation,in the detectionof CD8％,T#,CD4+ T# and CD8+ T#,BriCyte E6 achieved a low CV％ compared with FACSCanto Ⅱ and FC 500 (Friedman statistics are 16.720,11.840,15.760 and 15.430,P =0.000 2,0.027,0.000 4,0.000 4,respectively).In the detection of T％,CD4％,NK％,B％,NK#,B#,there was no significant difference among the 3 flow cytometers (Friedman statistics are 4.242,3.916,0.852,2.595,1.835 and 0.578,P =0.119 9,0.141 2,0.653 2,0.273 3,0.399 6,0.749 0,respectively).Conclusions The 3 flow cytometers have a good consistency in the detection of lymphocyte subsets.BriCyte E6 may be an alternative or complement of existing flow cytometers.

Objective To investigate the gene defects of a pedigree with inherited coagulation factor Ⅺ (FⅪ) deficiency by analyzing its phenotype and molecular genetic characteristics. Methods A pedigree with inherited FⅪ deficiency was enrolled in this study. The activated partial thromboplastin time (APTF), prothrombin time (PT), FⅪ activity (FⅪ: C) and FⅪ antigen (FⅪ: Ag) were determined for phenotype diagnosis. Fifteen exons and their flanks of F11 gene from the proband's genomic DNA were amplified by polymerase chain reaction (PCR), and the PCR products were directly sequenced to analyze the F11 gene mutation. The PCR products amplified from genomic DNA from the proband, her parents and 100 healthy donors were digested with restriction enzyme BssSI to exclude gene polymorphism and confirm the mutation site. The cleavage site in the signal peptide was predicted by the SignalP software. Results The values of APTT, PT, FⅪ: C and FⅪ: Ag of the proband were 69.5 s, 12.3 s, 2.6% and 2.5%, respectively, indicating that this case was cross-reacting material (CRM) negative. The same values of healthy controls were 35 s, 13 s, 100% and 100%, respectively. As compared with Genbank AY191837 sequence, four variants in F11 exons were found. G3733C heterozygous mutation in exon 2 causod Gly to Arg substitution at-1 amino acid position in signal peptide (G-1R). The G3733C mutation in exon 2 introduced a new BssSI enzyme digestion site. Further analysis of the 100 randomly collected DNA samples from the normal population excluded the possibility of G3733C as a polymorphism. CI6642T heterozygous mutation in exon 8 introduced a premature stop codon at 263 amino acid position (Q263Term). Conclusions G-1R mutation and Q263Term compound heterozygous mutation in F11 gene are the mechanism of FⅪ deficiency for the proband. G-1R mutation is a novel F11 gene mutation causing inherited FⅪ deficiency.