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Objective:To investigate the significance of multicolor flow cytometry (MFC) monitoring of minimal residual disease (MRD) in the course of allogeneic hematopoietic stem cell transplantation (allo-HSCT) after CD19-chimeric antigen receptor(CAR)-T cell immunotherapy for patients with refractory, relapsed B-cell acute lymphoblastic leukemia (r/r B-ALL).Methods:37 patients with r/r B-ALL admitted to Hebei Yanda Lu Daopei Hospital from January to July 2019, aged 15 (6, 19) years old, including 24 males and 13 females, were treated with CD19-CAR-T cell immunotherapy bridging allo-HSCT. MFC with cytoplasmic CD79a antibody to set up B-cell gates was used to monitor patients′ bone marrow (BM), cerebrospinal fluid (CSF), and tissue samples on day 0 (prior to the CAR-T cell immunotherapy), day 15, day 28 post CAR-T cell immunotherapy, and post transplantation.The MRD values of these samples were analyzed to evaluate the residual tumor cells and metastasis. The killing effect of the CAR-T cells was evaluated by the recovery of CD19+B cells before transplantation and the period between the timepoint when CD19+B cells was recovered and the timepoint when CAR-T cells were infused. Peripheral blood CAR-T cells were counted at different time points. Statistic analysis was performed by Kaplan-Meie assay and Log-rank test to analyze the difference of univariate cumulative survival.Results:(1)Among the 37 patients, 8 died and 29 survived. 5 patients relapsed after transplantation, of which 4 relapsed patients died and 1 survived. (2)MFC MRD negative remission rate of the death group was lower than that of the survival group at the following time points: post-CAR-T therapy and prior to transplantation (5/8 vs. 28/29, χ 2=7.540, P=0.006); day 15 of the CAR-T cell reinfusion (3/8 vs. 24/29, χ 2=6.512, P=0.011); day 28 of the reinfusion (3/8 vs. 276/29, χ 2=10.065, P=0.002). The probability of extramedullary MFC MRD positive tumor infiltration in the death group was higher than that in the survival group(7/8 vs. 14/29, χ 2=3.931, P=0.047). After CAR-T cell immunotherapy, the recovery period of CD19-positive cells in the death group, or the time for CAR-T cells to kill CD19-positive cells, was shorter than that in the survival group [42.00 days(30.00,49.00) vs. 55.00 days(41.50,73.50), Z=0.022, P=0.020]. Conclusion:The positive results of MRD by MFC at the following timepoints may predict unfavorable outcomes, such as post-CAR-T therapy and prior to transplantation, day 15 and 28 of the CAR-T cell immunotherapy, which may provide some guidance for clinical management.
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Objective:To investigate the effectiveness of detecting MC B/P in PB by FCM for EBV+PTLD screening.Methods:481 patients with fever and large lymph nodes after allogenic hematopoietic stem cell transplantation (ALLO-SCT) in Ludaopi Hospital from 2018 to 2019 were retrospectively analyzed. The results of post-transplantation days, viral load (EBV, CMV) and MCB/P were detected. To evaluate the value of MC B/P in the diagnosis of PTLD by the sensitivity, specificity, positive predictive value and negative predictive. Logistic regression was used to analyze the clinical influencing factors of EBV-associated PTLD. The median fellow-up time was 449 days (range: 184 days to 700 days).Results:The diagnosis of PTLD was established in 51 patients. 55 patients who detecting MC B/P by FCM were positive. There were significant differences between the PTLD negative and positive groups in lymph node enlargement, age, EBV, CMV, monoclonal B cells, and monoclonal plasma cells ( P<0.05). Monoclonal plasma, monoclonal B and days after transplantation are important relationship with the diagnosis of PTLD, which have good diagnostic value for EBV-associated PTLD. Conclusion:FCM screening peripheral blood MC B/P has good diagnostic performance for EBV-associated PTLD. Monoclonal B, monoclonal plasma and the number of days of PTLD after transplantation were correlated with EBV-associated PTLD.
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Objective:This study aims to analyze the counts (per kilogram of body weight) or percentages of transplanted lymphocyte subgroups in children with non-infectious pulmonary complications (NIPC) and air-leak syndrome (ALS) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and explore its significance in the progression of lung complications after transplantation.Methods:The patients with NIPC and ALS after allo-HSCT from January 2013 to December 2019 in Hebei Yanda Ludaopei Hospital were retrospectively studied and the influencing factors in the progress of NIPC after HSCT were statistically analyzed.Results:Of the 2026 children who received HSCT treatment, 59 patients (34 males and 25 females) developed NIPC, the probability was 2.9% (59/2 026), and the probability of combined ALS was 1.4% (28/206). The differences in the comparison between NIPC progressed to ALS group (ALS group) and failed to progress to ALS group (non-ALS group) in the patient′s age( P=0.028), disease condition before transplantation( P=0.022), NIPC onset time( P=0.004) were significant. The P values of the percentage of NKT-like cells in the bone marrow ( P=0.008) or peripheral stem cells ( P=0.003) accounted for the lymphocytes. CD4+CD25+dim cells in bone marrow ( P=0.029) or peripheral stem cells ( P=0.036) accounted for the CD4+lymphocytes and the ratio of CD4/CD8 in bone marrow( P=0.004) or peripheral stem cells ( P=0.020) were less than 0.05, which meant the differences in patients′ refusion cells were significant. In the binary logistic regression model, the percentage of bone marrow NKT-like cells to lymphocytes, the ratio of bone marrow CD4+/CD8+and the percentage of peripheral stem NK cells to lymphocytes were important risk factors for the progression of NIPC to ALS. The rest factors were excluded from the model (AUC=0.918, P<0.05). Conclusion:During allo-HSCT transplantation, a high proportion of NKT-like cell and NK cell levels, and a high CD4+/CD8+ratio in the infusion of donors with high immune tolerance have an important correlation with the progression of the NIPC.
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Objective To explore the clinical efficacy of omeprazole,levofloxacin and metronidazole in the treatment of chronic gastritis caused by Helicobacter pylori.Methods 100 patients with chronic gastritis were selected as the research subjects.In accordance with digital table method,100 patients were randomly divided into the observation group and control group.Each group had 50 patients.The observation group was given triple therapy with omeprazole,levofloxacin and metronidasole.The control group was only given omeprazole.Results The total effective rate of the observation group was 94％,which was significantly higher than 82％ of the control group (x2 =7.37,P ＜ 0.05).The negative rate of Hp in observation group was 90％,which was significantly higher than 72％ of the control group (x2 =8.12,P ＜ 0.05).Conclusion The clinical efficacy of triple therapy of omeprazole,levofloxacin and metronidazole was better than the single therapy of omeprazole,which has significant effect,less adverse reactions.