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Objective:To explore the performance and clinical application value of a rapid detection method for the hematopoietic stem/progenitor cell of peripheral blood using the automated hematology analyzer.Methods:Methodology validation and retrospective study. Collected sample from Fujian Medical University Union Hospital from January 2015 to December 2018, the peripheral blood of 4 patients with acute myeloid leukemia was first treated, and one healthy donor′s peripheral blood stem cell collection 5 times diluent, for the methodology validation. And the peripheral venous blood and 5-fold dilutions of peripheral blood stem cell collection, from 23 patients with hematopoietic stem cell transplantation and 22 healthy donors of allogeneic peripheral blood stem cell transplantation, used for consistency retrospective analysis. In the linear test, each of the peripheral blood and HPC collecting solutions from blood cell separator, which known CD34 +cell concentration, that was high-value samples for the expected upper limit (H) . Another low-value sample is normal saline (L) . According to the multiple proportion dilution, HPC was detected and regressed consistency test specimens were 126, EDTA-K 2 anticoagulant venous blood 78 and peripheral blood stem cell 48. Venous blood was collected at the same time, one tube of blood routine and HPC detection, the other tube flow cytometry (FCM) detection of CD34 +cells. Stem cell collection was diluted 5 times with sterilized saline and divided into two tubes. One tube was used to count whole blood cells and HPC, the other tube was used to detect CD34 +cells by FCM. The test results of the two instruments were compared, and the deviation was evaluated. Results:The background counting was 0×10 9/L and the carryover rate was 0.1%, conformed to the quality requirements of hematology analyzer, and the repeatability study imprecision ranged between 4.7%-18.8%. HPC of peripheral venous blood linear range (0-27.201×10 9/L), Stem cell collection was diluted 5 times linear range (0-0.878×10 9/L). The results of 126 samples detected by the hematology analyzer and FCM were compared. The correlation coefficient r2=0.960 1. When WBC>10×10 9/L, the results of the two instruments have a good consistency. The slope is between 0.95 and 1.05, and the relative bias is less than 30%. Conclusions:This study suggests that the hematology analyzer has a good linear range for detecting HPC, and has a good correlation with FCM. The hematology analyzer has the advantages of no pretreatment, convenient operation, a wide range of applications in detecting HPC specimens.
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Objective To analyze the molecular characteristics of qnrS-positive Escherichia coli ( E. coli) strains resistant to quinolone. Methods A total of 57 qnrS1-positive clinical isolates were collect-ed from Fujian Medical University Union Hospital. Plasmid-mediated quinolone resistance ( PMQR) genes [qnrA, qnrB, qnrC, qnrD, aac(6′)-Ib-cr, qepA and oqxAB] andβ-lactamase genes (blaCTX-M-1, blaCTX-M-2, blaCTX-M-8 , blaCTX-M-9 , blaSHV and blaTEM ) were detected by PCR and then sequenced. Agar dilution method was used to analyze the antimicrobial susceptibility of the qnrS1-positive strains. Phylogenetic analysis was conducted using PCR. Multilocus sequence typing ( MLST) was performed for phenotyping. Enterobacterial repetitive intergenic consensus-polymerase chain reaction ( ERIC-PCR) was used to evaluate the genetic sim-ilarity between those isolates. Transferability of the qnrS1 genes carried by the 57 strains was examined by conjugation test with the sodiumazide-resistant E. coli J53 as the recipient strain. Mutations in the quinolone resistance-determining regions ( QRDR) in those strains were analyzed by PCR. Results All of the qnrS1-positive E. coli strains showed high resistance to quinolones. PMQR genes were harbored by 14 (24. 6％) isolates. Extended spectrum β-lactamases (ESBLs)-producing isolates accounted for 68. 4％. Mutations in the QRDR of gyrA, gyrB, parC and parE genes were found in 56 (98. 2％) strains and the most frequent point mutations were S83L (89. 5％) in gyrA gene, S80I (54. 4％) in parC gene and P415V (28. 1％) in parE gene. The qnrS1 gene was successful transferred from 13 (22. 8％) isolates to E. coli J53 by conjuga-tion. Five plasmid incompatibility groups were detected. Phylogenetic analysis showed that there were 36 (63. 2％), 13 (22. 8％), 1 (1. 8％) and 7 (12. 3％) isolates belonging to groups A, B1, B2 and D, respectively. The 57 qnrS1-positive E. coli strains were assigned to 50 ERIC types and 39 sequence types ( ST) based on the results of ERIC-PCR and MLST. Conclusions Mutations in the QRDR in E. coli strains were associated with qnrS1 gene and might play a critical role in the dissemination of quinolone-resistant bacteria.
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Objective To evaluate the early diagnostic value of galactomannan ( GM ) test in bronchoalveolar lavage fluid ( BALF ) to invasive pulmonary aspergillosis ( IPA ) in patients with non-neutropenia.Methods A total of 199 patients with suspected IPA were enrolled in the Department of Critical Care Medicine and Respiration of the Union Hospital of Fujian Medical University from January 2016 to October 2017, these patients excluded neutropenia .The patients were divided into IPA group and non-IPA group according to the European Organization for Research and Treatment of Cancer /Infectious Diseases Mycoses Study Group ( EORTC/MSG ) consensus.BALF and serum GM tests were performed in both groups.SPSS 20.0 statistical software was used to analyze the results of both IPA group and non-IPA group.Results The GM index in BALF and serum of IPA group was 2.41 ±2.19 and 0.65 ±1.08 respectively.The former is higher than the latter , the difference was statistically significant ( u=8.27,P<0.0005 ) . The GM index in BALF and serum of non-IPA group was 0.37 ±0.33 and 0.2 ±0.15 respectively, compared with the IPA group, the difference were statistically significant (u=11.18 and 7.07, P<0.0005).When the cut-off of GM was equal or greater than 0.5, the sensitivity, positive predictive value and negative predictive value of GM test in BALF was 86.36％, 74.02％and 92.62％ respectively , were significantly higher than those in serum (31.8％, 67.74％and 73.21％respectively).The specificity of GM test in BALF was 84.96％, which was lower than that in serum (92.48％).When the cut-off of GM was equal or greater than 1.0, the sensitivity, positive predictive value and negative predictive value of GM test in BALF was 66.66％, 84.61％and 85.03％ respectively, significantly higher than those in serum ( 24.24％, 76.19％and 71.9％ respectively ) . The specificity was similar ( 95.98％vs 96.24％) . Conclusions GM test in BALF has a highly sensitivity and specificity in patients with non-neutropenia, better than that in serum.It has high value for early diagnosis of IPA.
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Objective To investigate the outpatient visitor′s B19 infection in Fuzhou area, study the correlation between B19 virus infection and clinical diseases.Methods The infection status of B19V IgM and IgG in 22 089 outpatient visitors in Fuzhou area from 2011 to 2016 has been retrospectively analyzed.The patients were divided into different groups according to sex,age,different pregnant outcomes, healthy people and hematopoietic system diseases.Results The positive rate in 22 089 patients of B19V IgM was 4.5%(998/22 089)and the IgG positive rate was 36.9%(8 155/22 089); The positive rate of B19V IgM in female patients(4.8%,546/11 374)was higher than male patients(4.2%,452/10 715)(χ2=4.333,P<0.05); The middle-aged and elderly patients IgG positive rate(53.6%,3 629/6 772;54.3%,1 542/2 838)were significantly higher than infants,children and young people(36.0%,989/2 747;25.4%,237/934;20.0%,1 758/8 797); The positive rate of IgM in adverse pregnancy outcomes (8.2%,20/245)was higher than normal pregnant women(3.3%,23/688)(χ2=9.548,P<0.05).In pancytopenia,thrombocytopenia and anemia patients, the positive rates of B19V IgG were 39.8%(165/415),38.1%(297/780)and 35.4%(81/226)respectively, all of which were higher than that in the healthy people(14.4%,78/543)(χ2=80.127,88.626,43.461; P<0.05).Conclusions The outpatient visitor′s infection rate of B19V in Fuzhou is high.B19V is a common virus who can lead to adverse pregnancy outcomes.What′s more, it also can lead to pancytopenia, thrombocytopenia, anemia or other related hematopoietic diseases.
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Objective To investigate the resistance mechanism of Carbapenem-Resistant Klebsiella oxytoca.Methods Car-bapenem-Resistant Klebsiellaoxytoca were collected from Fujian Medical University Union Hospital.The modified hodge test (MHT)was used for carbapenemase phenotype screening.The minimum inhibit concentration(MIC)was detected using agar dilution method for 1 7 drugs.PCR and DNA sequencing were used to detect commonβ-Lactamase genes and carbapene-mases genes.Conj ugation experiments demonstrated the transferability of the carbapenem-resistant determinants.Results 5 Carbapenem-Resistant Klebsiella oxytoca of 4 isolates were positive detected by MHT.Minimum inhibit concentration was detected by using agar dilution method for 17 drugs.More than 80% isolates were resistance to nine drugs.2 isolates conju-gated successfully of 5 Carbapenem-Resistant Klebsiella oxytoca Isolates.There were 2 isolates included carbapenemases gene (1 isolates were only IMP producers,1 isolate contained the IMP and KPC),3 isolates produce ESBLs gene.Conclution The due to CRE strains isolated from Fujian Medical University Union Hospital may be metallo-enzyme carbapenemase and KPC gene.And the isolate that produce two Carbapenem-Resistant gene had been found in this hospital.
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To combine more than 20 years of experience in clinical practice skills teaching of laboratory medicine, with the characteristics of laboratory medicine, the theory system of formative assessment has been constructed, to guide the clinical practice of the students.Based on the construction of network question bank, students make use of the network question bank self testing, to know whether they had got the stage goal, existing problems and future plan through self testing, so as to mobilize their enthusiasm and initiative to enhance their self-confidence.Under the formative assessment teaching system, students establish internship file information, including practice notes, weekly practice, group discussion, self testing results, the teacher and peer assessment information.Teachers set up QQ group, WeChat group with their students, the timely to get the question from students and to take appropriate measures improve teaching.Teachers had established and improved the long-term after graduation feedback mechanism, and formative assessment improved the teaching quality of the whole practice teaching benefits teachers as well as students.
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Objective:To explore the role and mechanism of the B7-H1 expressed by auto-keratinocytes in the intermingled skin grafting model in vitro(MELC). Methods:The intermingled skin grafting model(MELC) was established in vitro. Flow cytometry was performed to detect the expressions of B7-H1 in keratinocytes. The expressions of PD-1 in lymphocytes were measured at the same time. The levels of IL-10,Foxp3 and GATA-3 mRNA in lymphocytes were detected by Real-time quantitative PCR. Results: Through flow eytometry,in the MELC with auto-keratinocytes,the expression of B7-H1 in auto-keratinocytes and the PD-1 in lymphocytes were rising trend and the rising rate was in time-dependent manners(P<0. 01). RT-PCR assay indicated that the relative levels of IL-10, Foxp3,GATA-3mRNA expression were significant raised and the rising rate was in time-dependent manners (P<0. 01). Conclusion:In the intermingled skin grafting model,the auto-keratinocytes could express B7-H1 to enhance the expression of PD-1 in T cells. When B7-H1 combined with PD-1,the Th2 cells and Foxp3+Tregs were induced and suppressed the immune response in the intermingled skin grafting model.
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Objective To examine the prevalence of plasmid mediated quinolone resistance (PMQR) genes and their correlation with the genes encoding β-lactamases in E.coli isolates.Methods A total of 200 levofloxacin-and/or ciprofloxacin-resistant E.coli isolates were collected from Fujian Medical University Union Hospital during the period from July to December 2013.PCR method was used to screen these E.coli isolates for the presence of qnrA,qnrB,qnrC,qnrD,qnrS,aac(6')-Ib-cr,qepA,oqxAB genes,and the blaTEM,blasnv and blacTx-M genes in the PMQR positive strains.Agar dilution method was utilized to measure the antimicrobial susceptibility of PMQR-positive strains.Phylogenetic analysis was conducted by triplex PCR.Enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) was used to evaluate the genetic similarity between the PMQR-positive isolates.Results Of the 200 clinical isolates of E.coli,58 (29.0％)were PMQR-positive.And qnr,aac(6')-Ib-cr,oqxAB,and qepA genes were positive in 11 (5.5％),41 (20.5％),16 (8.0％),1 (0.5％) strains,respectively.The genes encoding CTX-M-1,CTX-M-9 and TEM type enzymes was positive in 32 (55.2％),17 (29.3％),and 1 (1.7％) of the PMQR-positive strains,respectively.The blasHv gene was not identified in any isolate.PMQR-positive strains were multi-drug resistant.Phylogenetic analysis indicated that 21 (36.2％),17 (29.3％),11 (19.0％),and 9 (15.5％) of the PMQR-positive strains belonged to group A,group D,group B2 and group B 1,respectively.ERIC-PCR suggested the PMQR-positive isolates belonged to 50 different types.Only one strain was non-typeable.Conclusions Most of the PMQR-related genes in E.coli are aac(6')-Ib-cr,qnr,and oqxAB in our hospital,which are highly relevant to β-1actamase genes.PMQR-positive strains may spread by way of non-clonal dissemination in our hospital.
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Cyr61/CCN1 is a secreted extracellular matrix ( ECM) protein, which has been shown to regulate a multitude of cellular responses.Many researches indicate that Cyr61 plays important roles in oncogenesis and development of tumor and is considered to be a novel potential oncogene.This review would provide a comprehensive summary about the roles of Cyr61 in the diagnosis and treatment of tumor.
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Objective To evaluate the value of Epstein-Barr virus (EBV) IgG antibody to EBV Rta protein(Rta-IgG),IgA antibody to EBV early antigen (EA-IgA) and IgA antibody to EBV viral capsid antigen(VCA-IgA) for nasopharyngeal carcinoma (NPC) diagnosis.Methods From May 2012 to July 2013,serum samples from 8 884 healthy donors,1 546 clinical screening patients and 155 NPC patients in Fujian Medical University Union Hospital were collected,and EBV Rta-IgG,EA-IgA and VCA-IgA were detected by ELISA.The ROC curve analysis and correlation analysis were performed to assess the value of Rta-IgG,EA-IgA and VCA-IgA for NPC diagnosis.The positive rate among three kinds of antibodies were compared with chi-square test.Results Positive rates of EBV Rta-IgG,VCA-IgA and EA-IgA in NPC patient group were 81.9％ (127/155),90.3％ (140/155) and 48.3％ (75/155),respectively,which were higher than those in clinical screening patient group and healthy donor group (x2 =1 538.6,479.3 and 643.3 respectively,P ＜ 0.01).The sensitivity of VCA-IgA (90.3％,140/155) and the specificity of EA-IgA (95.1％,9 915/10 430) were the highest in all groups.Further analysis showed that negative predictive value of EBV Rta-IgG,VCA-IgA and EA-IgA for NPC diagnosis were 99.7％ (9 826/9 854),99.8％ (8 168/8 183),99.2％ (9 915/9 995),respectively,suggesting that three anti-EBV antibodies showed very good negative predictive value for NPC diagnosis.Next,combined detection of three anti-EBV antibodies could improve the sensitivity (94.1％,146/155) and specificity (98.9％,10 469/10 585) for the NPC diagnosis.Conclusion The EBV Rta-IgG,VCA-IgA and EA-IgA all show the clinical value for NPC diagnosis,and combined detection for Rta-IgG,VCA-IgA and EA-IgA is more suitable to screen NPC and can improve the sensitivity and specificity of NPC diagnosis.
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In this study ,we detected the positive rate of anti‐HBs and anti‐HBc antibody among the subject population in Fujian Medical University Union Hospital ,and to evaluate different detection methods of anti‐HBc antibody .The positive rate of anti‐HBs and anti‐HBc antibody were detected by chemiluminescent microparticle immunoassay (CMIA) and one‐step com‐petitive enzyme‐linked immunosorbent assay (ELISA) from the year 2012 to 2013 .The subject population was divided into three groups :group 1 with the age of less than 2 years old ,group 2 with the age of 2‐20 years old ,and group 3 with the age of more than 20 years old .The positive rates of anti‐HBV antibody in the different groups were analyzed .Furthermore ,anti‐HBc antibody of 92 samples selected from the immunized population was detected by CMIA and three kinds of ELISA reagents . Meanwhile ,the detection of anti‐HBc antibody by the same ELISA reagent but different operating modes were performed in these samples .The highest positive rate of anti‐HBs antibody was detected in group 1 ,and there was no significance difference of positive rate between two detection methods of anti‐HBs antibody among three groups .The positive rate of anti‐HBc anti‐body using CMIA was significantly lower than those with ELISA among group 1 and 2 .Among the 92 samples ,the positive rate of anti‐HBc antibody was 2 .2% using CMIA .With three kinds of method of ELISA reagent ,the positive rate of anti‐HBc antibody were 79 .3% ,82 .6% and 94 .6% ,respectively ,and there was no statistical significance among the results of three ELISA reagents .Anti‐HBc was not detected from 19 samples using ELISA methods with different operating modes .It's con‐cluded that the anti‐HBs antibody declined with the increase of age ,and it is necessary to discriminate the specific population to strengthen immune system .The obviously higher positive rate of anti‐HBc antibody was found by ELISA in immunized popula‐tion than that by CM IA . Concerning on the false positive of ELISA , specimen sampling with one specific test item or the CMIA method was recommended to detect the anti‐HBc antibody .
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Objective To investigate the effect of Intedeukin-18 (IL-18) on Th1/Th2 balance and its antitumor mechanism in C57BL/6 mice Lewis lung cancer model. Methods 24 C57BL/6 mice were randomly divided into three equal groups: group A(IL-18 injec-tion group, n = 8), group B (Lewis lung cancer model, n = 8) and group C (normal control group, n = 8). The Lewis lung cancer cells were cultured and implanted subcutaneously into the group A and group B. IL-18 and NS were given to group A and B respectively by intrap-eritoneal injection on the 7th day (once every day, 7 times altogether), but group C was not given any treatment. Enzyme-linked immunosor-bent assay (ELISA) was used to detect the Th1/Th2 cytokines. Health status in all the animals was evaluated; the volume and weight ofsubcutaneous tumors were measured. Results The concentration of IFN-γ in group A and C were significantly higher than those in group B (P <0.05), and the concentration of IL-4 in group A and C were significantly lower than those in group B (P<0.05), but there was no significant difference between group A and C (P>0.05). The tumor growth inhibitory rate was 75%. Conclusion IL-18 can effectively induced IFN-γ and inhibit IL-4 production, regulate Th1/Th2 balance in the C57BL/6 mice Lewis lung cancer model, and elicit the antitu-mor immunity of the host, which could obviously inhibit the growth of tumor cells and decelerate the proliferation of tumor cells.
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Objective:To investigate the possible role of NKT cells in unexplained recurrent spontaneous abortion by measuring the NKT cell numbers,maturity and cytokine secretion of the placenta of mice with unexplained recurrent spontaneous abortion.Methods:Normal pregnancy model in hybrid by feeding CBA / J and BABL/C in a cage,and the model of unexplained recurrent spontaneous abortion was established by feeding CBA / J and DBA2/J in a cage.The number of NKT and CD3~+T cells was determined by flow-cytometry;Th1/Th2-relative cytokines were assayed by ELISA and T-bet expression was determinded by fluorescence quantitative PCR.Results:There was not significant change of CD3~+ T cells when compared between normal pregnancy and unexplained recurrent spontaneous abortion group (P＞0.05).In the course of normal pregnancy,the IFN-γ secreted by placenta lymphocytes decreased gradually,accompanied by the decline of NKT cell number and the proportion of mature cells;whereas in the course of unexplained recurrent spontaneous abortion,it was on the opposite.There was significant difference of T-bet mRNA expression between the two groups.T-bet mRNA expression was related to the proportion of mature NKT cells or placenta IFN-γ secretion by lymphocytes.Conclusion:Unexplained recurrent spontaneous abortion may be related to NKT cells disorders,NKT cells are of low-mature proportion and inadequate secretion of IFN-γ during early pregnancy,whereas are shown high-mature proportion and excessive secretion of IFN-γ during latter pregnancy;the anomaly of T-bet mRNA expression may be one of the factors leading to NKT cells disorder.
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S100B protein has a higher sensitivity to central nervous system(CNS),which can be used in clinical and prognostic prediction of various forms and different degrees of CNS injury,even in the judgment of brain death.The measurement of S100B protein and prediction of secondary brain injtry enable physicians to select an appropriate opportunity for therapy intervention,which will be a focus in the future studies.S100B protein is a potential protein biomarker that has clinical prospect.