ABSTRACT
Chimeric Antigen Receptor-modified(CAR)-T cells have become a new star living cell "drug" in the field of tumor immunotherapy, and five drugs have been available to the public since 2017. The favourable, rapid and efficient interaction of CAR-T therapy from bench to bedside has solved many new scientific problems, which remarkably expands the application field of multi-parameter flow cytometry (MFC). MFC participates the whole process of CAR-T preparation, functional evaluation, quality control, in vivo continuation evaluation, clinical efficacy and toxicity monitoring. New clinical problems caused by high-precision targeted therapy of CAR-T cells pose new challenges in using MFC for accurate tumor immunophenotyping, minimal residual disease monitoring, as well as the demands of comprehensively evaluation of the systematic immune function in different disease stages. In the era of targeted therapy, timely communication between laboratories and clinics is particularly important for obtaining accurate MFC results, which assists clinical individualized diagnosis and treatment.
ABSTRACT
Objective@#To probe the prognostic value of consolidation chemotherapy in non-favorable acute myeloid leukemia (AML) patients who were candidates for allogeneic hematopoietic stem cell transplantation (allo-HSCT) with first complete remission (CR1) and negative minimal residual disease (MRD-) .@*Methods@#A retrospective analysis was conducted on 155 patients with non-favorable AML who received allo-HSCT in CR1/MRD- from January 2010 to March 2019. The survival data were compared between patients who received and those not received pre-transplant consolidation chemotherapy.@*Results@#A total of 102 patients received pre-transplant consolidation chemotherapy (consolidation group) , and 53 cases directly proceeded to allo-HSCT when CR1/MRD- was achieved (nonconsolidation group) . The median ages were 39 (18-56) years old and 38 (19-67) years old, respectively. Five-year post-transplant overall survival [ (59.3±7.5) % vs (62.2±6.9) %, P=0.919] and relapse-free survival [ (53.0±8.9) % vs (61.6±7.0) %, P=0.936] were not significantly different between the two groups (consolidation vs nonconsolidation) . There was a weak relationship between consolidation therapy and cumulative incidence of relapse [consolidation: (21.9±5.4) % vs nonconsolidation: (18.3±6.0) %, P=0.942], as well as non-relapse mortality [consolidation: (22.4±4.3) % vs nonconsolidation: (28.4±6.5) %,P=0.464]. Multivariate analysis indicated that pre-transplant consolidation and the consolidation courses (< 2 vs ≥2 courses) did not have an impact on allo-HSCT outcomes.@*Conclusion@#Allo-HSCT for candidate patients without further consolidation when CR1/MRD- was attained was feasible.
ABSTRACT
Objective@#To compare the difference of efficacy between traditional Hyper-CVAD/MA regimen and the adolescents inspired chemotherapy regimen, CH ALL-01, in treatment of adult Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL) .@*Methods@#In this study we retrospectively analyzed 158 Ph+ ALL patients receiving Hyper-CVAD/MA regimen (n=63) or CHALL-01 regimen (n=95) in our center and Changzheng hospital from January 2007 to December 2017, excluding patients with chronic myeloid leukemia in blast crisis. Tyrosine kinase inhibitor (TKI) was administered during induction and consolidation chemotherapy. Patients who underwent hematopoietic stem cell transplantation received TKI as maintenance therapy.@*Results@#Of them, 91.1% (144/158) patients achieved complete remission (CR) after 1-2 courses of induction. CR rate was 90.5% (57/63) for patients in Hyper-CVAD/MA group and 91.6% (87/95) for patients in CHALL-01 group. There was no difference in CR rates between the two groups (χ2=0.057, P=0.811) . The last follow-up was June 2018. A cohort of 134 CR patients could be used for further analysis, among them, 53 patients received Hyper-CVAD/MA regimen and other 81 patients received CHALL-01 regimen. The molecular remission rates were significantly higher in CHALL-01 group (complete molecular response: 44.4%vs 22.6%; major molecular response: 9.9% vs 18.9%) (χ2=7.216, P=0.027) . For the patients in Hyper-CVAD/MA group, the 4-year overall survival (OS) was 44.81% (95%CI: 30.80%-57.86%) and the 4-year disease free survival (DFS) was 37.95% (95%CI: 24.87%-50.93%) . For patients received CHALL-01 regimen, the 4-year OS was 55.63% (95%CI: 39.07%-69.36%) (P=0.037) and 4 year DFS was 49.06% (95%CI: 34.24%-62.29%) (P=0.015) , while there was no significant difference in 4 year cumulative incidence of relapse (CIR) (P=0.328) or cumulative incidence of nonrelapse mortality (CI-NRM) (P=0.138) . The rate of pulmonary infection was lower in patients received CHALL-01 regimen compared with patients received Hyper-CVAD regimen (43.4% vs 67.9%, χ2=7.908, P=0.005) .@*Conclusions@#Outcome with CHALL-01 regimen appeared better than that with the Hyper-CVAD/MA regimen in Ph+ ALL, which has lower incidence of pulmonary infection, higher molecular remission rate and better OS and DFS.
ABSTRACT
Objective@#To investigate the relationship between donor chimerism and relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT) .@*Methods@#The clinical data of 105 patients with acute myeloid leukemia (AML) who underwent allo-HSCT and recurrence-free survival>90 days from January 2010 to January 2019 were retrospectively analyzed. The bone marrow samples were collected at 15, 30, 60, 90, 180, 270, 360 days after transplantation. Donor chimerism was detected by single nucleotide polymorphism (SNP) -PCR.@*Results@#Of the 105 patients, 43 cases were male and 62 cases were female, with a median age of 38 (16-60) years. Till April 2019, the median follow-up was 843 (94-3 261) days. Ninety days after transplantation, 18 cases relapsed, 33 cases died, and 72 cases survived. The 3-year overall survival (OS) rate was (66.8±5.1) %, and the recurrence-free survival (RFS) rate was (65.1±5.0) %. Pre-transplant disease status, pre-transplant minimal residual disease (MRD) , and 90 day post-transplantation chimerism were independent risk factors related to RFS. The risk of recurrence was significantly increased in patients with a donor chimerism rate ≤97.24% at 90 days after transplantation[HR=6.921 (95%CI 2.669-17.950) , P<0.001], which was considered as a sign of early relapse.@*Conclusion@#SNP-PCR is an applicable method for detecting donor chimerism in patients after allo-HSCT. Chimerism rate equal or less than 97.24% at 90 days after transplantation predicts a higher risk of relapse.
ABSTRACT
Objective By a sequencing panel consisting of 50 targeted genes, aiming at depicting the molecular landscape of ET, PV, and PMF, which are three major subtypes of MPN, to provide valuable information in the diagnosis and prognosis of MPN.Methods A retrospective study was conducted of 53 patients from Huashan hospital and Changhai hospital. All patients were diagnosed in accordance with the 2016 WHO diagnostic criteria for MPN, including 31 cases of ET(11 males, 20 females, median age 55 years), 17 cases of PV(12 males, 5 females, median age 65 years), and 5 cases of PMF(4 males, 1 females, median age 67 years), and underwent next-generation of DNA sequencing of their bone marrow or blood samples. The genetic analyses were performed on bone marrow or peripheral blood. Referring to COSMIC, dbSNP, Clinvar and other public databases, we analyzed the sequencing data, and elucidated the mutation profile of MPN patients, combining with their clinic information. Results In addition to the typical JAK2, CALR, and MPL mutations, pathogenic mutations in other 11 genes were detected, as well as 4 SNPs that confer individual susceptibility to MPNs (rs4858647, rs9376092, rs58270997, rs621940). The average rate of mutated genes was 2.3 genes per patient. In all patients (53 cases), the mutated genes detected were TET2, EZH2, ASXL1, MIR662, SF3B1, BARD1, DNMT3A, KIT, RUNX1, TP53, NRAS according to their mutational frequency. Conclusions Applying next-generation sequencing technology, multi-gene sequencing of a bunch of typical BCR-ABL-negative MPN patients can be performed at one time within 2 working days, and pathogenic mutations other than JAK2, CALR, MPL can be found, which has a bright prospection in clinic.
ABSTRACT
Objective To investigate the diagnosis, treatment and prognosis of acute promyelocytic leukemia (APL) with NPM_RARα fusion gene positive. Methods One APL patient with NPM_RARα fusion gene positive who was diagnosed by using morphology, immunology, cytogenetics, molecular biology and multiplex fluorescence in situ hybridization in Changhai Hospital in November 2014 was retrospectively analyzed, and the patient was induced with retinoic acid and treated with DA (daunorubicin + cytarabine) regimen, followed by 4 courses of cytarabine consolidation therapy. Results Abnormal promyelocyte accounted for 0.64 by morphology. And the group of cells expressed myeloperoxidase (MPO), CD13, CD15, CD117, and CD7, CD11c, CD79a, CD123 weakly expressed or not by immunophenotype analysis; karyotype analysis showed 45, XY, t(5;17), 7p-,-16[8]/46, idem,+20[5]/45, idem,-8,+20[2]/46, XY[5]; the fusion gene screening showed that the expression level of NPM_RARα was 416.98% compared with that of APL; molecular complete remission was obtained after the consolidation therapy, but the patient relapsed after 34 months. Finally, the patient died of abnormal coagulation and respiratory failure, with overall survival of 35 months. Conclusion APL with NPM_RARα fusion gene positive is a rare type of acute leukemia, and the main treatment method is retinoic acid combined with myeloid chemotherapy regimen, which has a favorable efficacy but a poor prognosis.
ABSTRACT
Chimeric antigen receptor modified T cell(CAR-T)immunotherapy has achieved remarkable success in the treatment of hematological malignancies,especially for relapsed/refractory acute B lymphocytic leukemia(B-ALL)leukemia patients,most of whom could obtain complete remissions or partial remissions.However,clinical studies also found a as high as 50%total recurrence rate of these patients,in which more than 20% patients relapsed with tumor cells not expressing the primary CAR-T-targeted CD molecules.Increased relapses with negative targeted-molecules or even lost primary abnormal tumor gene, suggesting that immune escape and even clonal evolution events increased rapidly under the high -precision CAR-T immunotherapy pressure.Monitoring of minimal residual disease(MRD)is important to assess the efficacy and find recurrence trends early in a timely manner to guide further clinical intervention.The new features of disease recurrence after CAR-T treatment have put forward higher requirements for traditional and emerging MRD detection methods so as to ensure their effectiveness in the CAR-T treatment era.
ABSTRACT
Objective To analyze the application of quality control cycle (QCC) in reducing the false negative rate of minimal residual disease (MRD) of flow cytometry in patients with acute myeloid leukemia (AML). Methods In AML patients with abnormal fusion gene detected in hematology laboratory of Changhai Hospital during the year of 2014, the prevalence of AML-MRD detected both by flow cytometry (FCM) and real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) were analyzed retrospectively. The possible causes of false negative rate of flow cytometric MRD referring to PCR were further deeply analyzed, and the improvement measures were adapted from January 2015 to December 2015 and further judged all according to the QCC methods. Results Pareto diagram showed that the dilution and coagulation of the specimen, the improper analysis strategy and the incomplete combination of the MRD index [composition ratio:83.3 % (60/72)] were the main factors leading to the leakage of FCM MRD in 2014. The QCC group devised measures to reduce the dilution probability of bone marrow and develop a standard operating procedures (SOP) for sampling and testing, strengthen the maintenance of the flow instrument and more importantly, focused on optimizing the antibody panels and gated strategies referring to the current two main kinds of MRD detection combination modes on the basis of the latest advances published in 2015. Finally, the undetected rate of AML-MRD was reduced by FCM from 14.8 % (72/486) in 2014 to 2.6 % (16/620) in 2015. Conclusions The QCC can effectively reduce the leakage rate of flow cytometric AML MRD, improve the ability of laboratory quality control and the ability to solve problems. Solving problems with QCC is thus worthy of being popularized.
ABSTRACT
Objective@#To study the expression of CD123 in bone marrow (BM) blasts of acute myeloid leukemia (AML) patients to explore the relationship between CD123 expression and therapeutic response and prognosis.@*Methods@#This study retrospectively analyzed expression and distribution of CD123 in BM blasts in 137 cases of newly diagnosed AML (excluded M3) , CD123 detected by flow cytometry≥20% was defined as positive, including 84 CD123+ AML and 53 CD123- AML, efficacy and prognosis were compared between the two groups.@*Results@#① Among 137 patients, 84 were in group CD123+ (61.3%) , and 53 in group CD123- (38.7%) . All 137 patients were classified into risk groups based on cytogenetic and molecular biology abnormalities. No significant differences were seen between the three risk groups with regard to their CD123 levels (χ2=0.861, P=0.650) . Compared with CD123- group, the CD123+ group had higher WBC[47.7 (1.0-264.0) vs 22.4 (0.7-211.0) , z=-2.592, P=0.010]. ② The rates of first complete remission (CR1) and recurrence of CD123+ group were 54.8% (46/84) and 50.8% (32/63) , respectively; and CD123- group were 73.6% (39/53) and 41.7% (20/48) , respectively. There was significant difference of CR1 between the two groups (χ2=5.121, P=0.027) , whereas no significant difference of the recurrence rate (χ2=0.911, P=0.340) . ③ The median dutations of OS between CD123+ group and CD123- group were 20.0 (95%CI 13.1-26.9) months vs 44.0 (95%CI 23.6-47.3) months, respectively (χ2=5.874, P=0.015) ; The median durations of DFS were 7.8 (95%CI 1.4-14.1) months vs 18.6 (95%CI 0-39.7) months, respectively, no differences were observed between the two groups (χ2=2.939, P=0.086) . ④ CD123 retained an adverse prognosis value on DFS and OS within the intermediate group and patients ≤ 50 years older.@*Conclusions@#CD123 widely expressed in AML patients, which was an independent risk factor for CR1 and OS, which implicating its important role in evaluating the induction chemotherapy response and prognosis of AML.
ABSTRACT
Translational medicine and personalized medicine are becoming the new power in modem medical development.Laboratory Developed Tests (LDTs) based on molecular biology and bioinformatics technology play a vital role in the development process of translational medicine and personalized medicine.Nowadays,LDTs consist of a broad range of in vitro diagnositc tests performed to analyze nucleic acid,chromosomes,proteins,certain metabolites and cell surface molecules using immunological technique,cytogenetic or molecular methods or a combination of these methods.These LDTs are used to detect heritable or acquired disease-related genotypes,mutations,or phenotypes,and also used to classify the pathological changes of cells for clinical purposes.New generation LDTs,present some unique regulatory questions still remain to be discussed.Clinicians should capture the opportunity to establish and continuously improve the detection platforms of LDTs,the quality control system and management standards.On this basis,transformation bridges between scientific research and clinical application will be truly built.
ABSTRACT
Objective To investigate the detection methods of atypical bcr-abl rearrangement with b3a3 fusion transcript,and to describe the characteristics of this fusion gene.Methods Karyotype analysis,FISH and RT-PCR were applied to detect the break point of bcr-abl fusion gene in a patient who was diagnosed as acute lymphoblastic leukemia.Results The karyotype of the patient was expressed as 45,XY,-7,t(9;22)(q34;q1 1).The translocation event in chromosome 9 and 22 could be successfully detected by FISH,and a rare bcr-abl rearrangement with b3a3 fusion transcript was detected by RT-PCR with specific primers.Conclusions The rare e14a3 (b3a3) fusion of bcr-abl gene is present in this patient.Clinical laboratories using commercial kits that do not cover such rare fusions are likely to generate false result,thereby declaring combination of various methods to detect fusion genes is necessary.More studies are needed to explore the function and significance of rare bcr-abl fusion genes.
ABSTRACT
<p><b>OBJECTIVE</b>To explore the effects and possible mechanisms of decitabine on Molt4 in vitro.</p><p><b>METHODS</b>Effects of decitabine on cells proliferation were detected by using CCK-8, the apoptosis by Annexin V-FITC, cell cycles by propidium iodide-FACS. Discrepancy genes were screened by RNA-seq technique. The CpG methylation of lactoferrin (LTF) gene in Molt4 cells were identified by Bisulfite sequencing PCR (BSP). The expression of LTF mRNA in Molt4 by RT-PCR and LTF protein expression were analyzed by Western blot.</p><p><b>RESULTS</b>Decitabine effectively inhibited proliferation and induced apoptosis for Molt4 cells by an time- and dose-dependent manners. Cell cycles were arrested at the G₀/G₁ phase. The promoter methylation degree of LTF gene in Molt4 cells was 72.3% before decitabine treatment and decreased to 45.0% after treatment with 0.50 μmol/L decitabine for 72 h. After the reduction of methylation, expression of its mRNA and protein increased, meanwhile caspase 3 and caspase 9 protein expression levels increased.</p><p><b>CONCLUSION</b>The demethylating drug decitabine can induce apoptosis, detain cell cycle at phase G₀/G₁, inhibit proliferation and up-regulate LTF gene expression in Molt4 cells. LTF may become a new target for acute T lymphoblastic leukemia.</p>
Subject(s)
Humans , Antimetabolites, Antineoplastic , Apoptosis , Azacitidine , Caspase 3 , Cell Cycle , Cell Line, Tumor , Cell Proliferation , DNA Methylation , Lactoferrin , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma , Promoter Regions, GeneticABSTRACT
<p><b>OBJECTIVE</b>To improve the MigR1-CD19-CAR (chimeric antigen receptor) that contains a single chain variable region (scFv) which targeted to CD19 through a retroviral vector transduction efficiency of T-lymphocytes.</p><p><b>METHODS</b>Insert the CD19-CAR fragment into the retroviral vector (MigR1) through recombinant DNA technology, after transfecting plat-A packaging cell lines, viral supernatant was collected to transduce K562 cell line and activated human T-lymphocytes. We used flow cytometry to determine the transduction efficiency and RT-PCR to confirm the transcription of CD19-CAR gene. The ability of the transduced T cells to produce IFN-γ and TNF-α in a CD19-specific manner was measured in an enzyme-linked immunosorbent (ELISA) assay.</p><p><b>RESULTS</b>(1)Using MigR1-CD19-CAR retroviral vector to produce the high titer retrovirus. (2)MigR1-CD19-CAR transduction efficiency of K562 cell line was significantly higher than human T-lymphocytes (P<0.01). (3)120 min centrifugation could significantly improve transduction efficiency of T-lymphocytes to (54.5±14.6)%. (4)Transduction efficiency could be improved by deciding transduce time according to T-lymphocytes proliferation fold in vitro individually, and the highest transduction efficiency in the study was 69.3%. The CD19-CAR gene sequence was transcripted specificly with high efficiency. (5) IFN-γ and TNF-α released by CD19-CAR transduced T-lymphocytes significantly increased to (13 230±1 543) pg/ml and (4 217±211) pg/ml when coculture with CD19-K562 cells.</p><p><b>CONCLUSION</b>We have successfully constructed a second generation CAR which targeted to CD19 through a retroviral vector called MigR1 (MigR1-CD19-CAR). Deciding transduce time according to T-lymphocytes proliferation fold in vitro individually and 120 min centrifugation could improve the CAR transduction efficiency of T-lymphocytes. RT-PCR confirmed that the CD19-CAR gene was specificly transcripted with high efficiency. IFN-γ and TNF-α released by CD19-CAR transduced T-lymphocytes significantly increased when activated by target cells.</p>
Subject(s)
Humans , Antigens, CD19 , Cell Proliferation , Flow Cytometry , Genetic Vectors , K562 Cells , Recoverin , Retroviridae , T-Lymphocytes , TransfectionABSTRACT
Objective To construct a MigR1-CD19 recombinant vector which contains CD19 gene, and to establish a CD19-K562 cell line over-expressing stably CD19 gene and its subcutaneous xenograft model in NOD-SCID mouse.Methods The CD19 gene was inserted into the retroviral vector (MigR1) through recombinant DNA technology after transfection into Plat-A packaging cells, and viral supernatant was collected to transduce K562 cell line repeatedly to obtain stable transduction CD19-K562 cell line.Flow cytometry was used to determine the transduction efficiency and RT-PCR was used to confirmed CD19 gene expression.Cell proliferation and apoptosis were detected by cell count and Annexin V/PI, respectively.Then the subcutaneous xenograft subtype of CD19-K562-a cell line was constructed through subcutaneous inoculation and was cultured in vitro and in vivo.Then its subcutaneous xenograft model in NOD-SCID mouse was established.The characteristics of CD19-K562-a cells were detected by RT-PCR, Wright staining and immunohistochemistry.Results MigR1-CD19 recombinant vector was successfully constructed, and the CD19 positive efficiency of K562 cell line was (99.80±0.17) ％ through retrovirus centrifugation transduction.The transduction and passage had no effects on proliferation and apoptosis of CD19-K562 cells.The CD19-K562-a cell line was constructed after CD19-K562 cells were injected subcutaneously and were passaged in vitro and in vivo.The CD19 positive efficiency of the xenograft subtype CD19-K562-a cell line was (99.78± 0.04) ％.CD19-K562-a and CD19-K562 cells were in an undifferentiated state.NOD-SCID subcutaneous xenografts were established through subcutaneous inoculation of CD19-K562-a cells.CD19 in the CD19-K562-a subcutaneous xenografts was positive, while it was negative in its counterparts K562 cells.Conclusion The CD19-K562 cell line over-expressing CD19 gene and its subcutaneous xenograft model in NOD-SCID mouse are successfully established.
ABSTRACT
Objective To investigate the value of serum IgG4 in diagnosis of IgG4-RD and in differentiation from rheumatic diseases.Methods Total of 23 patients with IgG4-RD and 502 patients with rheumatic diseases were enrolled,who presented at Changhai Hospital in 2010 to 2011.In the study,rheumatic diseases were categorized into groups of Sj(o)gren syndrome (n =26),ankylosing spondylitis (n-50),systemic sclerosis (n =3),rhcumatoid arthritis (RA,n =125),mixed connective tissue disease (n =15),systemic lupus erythematosus (SLE,n =212),adult onset still disease (n =20),Behcet syndrome (n =17),polymyositis (n =12),dermatomyositis (n =12),polymyalgiarheumatica (n =10).Serum IgG and IgG4 levels were measured by a rate nephelometer assay.The ROC curves were constructed to identify the optimal serum IgG4 cutoff value for diagnosing IgG4-RD and evaluate its sensitivity and specificity.Results The mean levels of serum lgG4 in the group with IgG4-RD were 11.4(5.0-14.8) g/L.In about 95.6％ IgG4-RD patients,the serum IgG4 level was higher than ＞ 1.4 g/L and other rheumatic diseases (U values were 6.0,21.0,0,58.5,0,9.0,3.0,4.0,0,3.0,3.5,P ＜0.01).The levels of serum IgG4 with RA was 0.6(0.3-1.2) g/L,the levels of serum IgG4 with SLE was 0.2 (0.1-0.4) g/L.There were statistical differences between RA and SLE (U value was 5847,P ＜ 0.01).At the same time,some patients with other rheumatic diseases were found serum IgG4 level higher than ＞ 1.4 g/L,which was about 10％ in the patients whith RA,ankylosing spondylitis,adult onset still disease and polymyalgiarheumatica.According to the ROC constructed the cut off value in present study was 2.2 g/L,and sensitivity and specificity were 95.7％ and 97.4％,respectively.Area under the cerve (AUC) was 0.995.There were no significant differences between the sensitivity and specificity values obtained with a cutoff value of 2.2 g/L.In patients with other rheumatic diseases,the ratio of high serum IgG4 level (＞ 2.2 g/L) were declined obviously,except polymyalgiarheumatica,it was less than 10％.For differentiation from rheumatic diseases specificity values were higher.Conclusions The cut off value of 2.2 g/L is useful for diagnosing IgG4-RD,and in differentiation from rheumatic diseases.The high serum IgG4 concentrations are not specific to IgG4-RD.The cut off value of 2.2 g/L is better to diagnose IgG4-RD,and contributes to the differential diagnosis of IgG4-RD and other rheumatic diseases,but it needs to be further confirmed in clinical practice.
ABSTRACT
Disease-associated autoantibodies (AAB) are important for the diagnosis of respective autoimmune diseases (AID).Autoantibodies can also be used for monitoring of response to therapy and for prognostic purpose.However,significant biological heterogeneity of autoantibody response,the difficulty in simultaneously improving detection sensitivity and specificity of autoantibodies and the lack of standardization in detection methods lead to limitations in its clinical applications and some difficulties in explaining the test results.It is important to search for novel autoantibodies in sera,to establish and standardize automated detection platforms with good quality and to perform well-designed clinical evaluation in the future research and clinical applications of autoantibodies.
ABSTRACT
microRNA(miR) in body fluids shows good stability and the change in its expression profile and level is associated with cancer and other diseases.Thus,circulating miR has been proposed to be an useful biomarker in diagnosis of the onset,prognosis and risk of diseases.Although great discoveries have been obtained on all aspects of miR,there are some shortcomings existing in the present studies,such as inconsistent specimens,diverse detection methods,lack of standardization on data analysis and other defects,resulting in poor reproducibility of data and inconsistent clinical conclusion,that make circulating miR difficult to be used in clinical laboratory testing.So lots of work is needed for us to implement,such as standardizing sample selection,improving detection reagents and methods,and normalizing test results before we can ultimately establish a fast,easy,low-cost detection of circulating miR in the clinical laboratory.
ABSTRACT
Objective To explore the possible negative interference of circulating cardiac troponin Ⅰ(cTnI) autoantibody on the immunoassay of cTnI in five commonly used cTnI detection systems. Methods Thirteen patients with positive cTnI autoantibodies in their serum samples were firstly screened and selected from 121acute myocardial infarction (AMI) patients using ELISA assay. The serum cTnI values and their recovery rates were then carefully measured and analyzed. Results cTnI values in these 13 samples showed amazing difference in the five detection systems, demonstrating various degrees of pseudo-drop, or even false-negative. One sample with low recovery was detected in Access-2 system. One sample with low recovery as well as one sample with moderate recovery were detected in Architect i2000 (Abbott). Two samples with moderate recovery and one sample with low recovery were detected in Axsym(Abbott). Three samples with moderate recovery and two samples with low recovery were detected in Dimension X Pand (Dade Behring)and one sample with moderate recovery together with four samples with low recovery were detected in Vidas (Biomerieux). And the serum levels of autoantibodies (A450) positively correlated with the degrees of their negative interference for the detection of cTnI. The R2 and P values on each system were 0. 841 (P <0. 01)vs Access-2, 0. 808 (P < 0. 01) vs Architect i2000 (Abbott), 0. 772 (P < 0. 01) vs Axsym (Abbott), 0. 707 (P < 0. 01) vs Dimension X Pand (Dade Behring) and 0. 424 (P < 0. 05) vs Vidas (Biomerieux), respectively. Conclusion Circulating autoantibodies of cTnI can induce considerable negative interference in all the 5 commonly used cTnI detection systems, which might then lead to incorrect judgments of the obtained results of cTnI in daily clinical work.
ABSTRACT
Substances that might potentially alter the measurable concentration of the analyte or alter the binding ability of detection antibody can lead to immunoassay interference. Endogenous interferences consist of autoantibodies, heterophiles antibodies, human anti-animal antibodies (HAAA) and some binding proteins. Lipidemia, cross-reactivity, pre-analytical variation, matrix and different detection equipments may also affect immunoassay. These interfering substances may cause falsely increased or decreased concentration in many analytes, including hormones, tumor markers, drugs, cardiac tropanin and microbial serology, thus it will affect the diagnosis of patients and the evaluation of therapeutic strategies Laboratorians and physicians should both be aware of the potential interference in immunoassays and communicate closely when any clinical discordance between the clinical and the laboratory data appears, avoiding a subsequent wrong diagnosis and unwarranted treatment. In this case, an alternative assay or measurement is needed for the potential correct results.
ABSTRACT
Objective To investigate the effect of aquaporin 5(AQP5)gene to the differentiation and development of bone marrow-deriued dendritic cells(BMDC).Methotis The DCs obtained from the bone marrow of the AQP5-/- and the wild type(WT)mice were cultured and induced to maturation by LPS.The phenotype of the two types DCs were assayed by FACS,and the phagocytosis ability Was assayed by co-incubating with FITC-conjucted ovalbumin.Results Contrast to the DCs from the WT mice,the DCs from the A9P5-/-mice showed reducing positive rate of expression of the co-stimulating molecular such as CD40,CD80.CD86.And the phagoeytosis ability of DCs from the AOP5-/-mice was lower than that of the WT mice.The relationship of the phagncytosis rate to the time from the AOP5-/- mice was different from that of the WT mice.The stimulating ability of the AOP5-/-DCs was lower than that of the WT mice.Conclusion Water transmembrane moving mediated by AQP5 iS very important to the normal function of DCs.The certain mechanism of the signal moleeular is to be determined. Aquaporin 5;Dendritic cell;Gene knockout;Co-stimulating molecular