Effect of Hemoglobin on Efficacy of CAR-T Therapy in Patients with Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of hemoglobin (Hb) on the efficacy of chimeric antigen receptor T cell therapy (CAR-T) in patients with multiple myeloma (MM).@*METHODS@#From June 2017 to December 2020, 76 MM patients who received CAR-T therapy in the Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, with complete clinical data and evaluable efficacy, were selected as the research objects. According to the receiver operating characteristic (ROC) curve, the best cut-off value was obtained. The patients were divided into groups on the basis of Hb 105.5 g/L as the cut-off value. The age, sex, serum calcium, β2-microglobulin, serum creatinine, lactate dehydrogenase (LDH), and the influencing factors of CAR-T treatment efficacy in MM patients were analyzed.@*RESULTS@#Hb was an influencing factor of efficacy. Univariate analysis showed that Hb, LDH, and albumin affected the efficacy of CAR-T therapy. Multivariate analysis showed that Hb ( OR=1.039, 95% CI: 1.002-1.078) and LDH ( OR=1.014, 95% CI: 1.000-1.027) were the influencing factors for the efficacy of CAR-T therapy.@*CONCLUSION@#The efficacy of CAR-T therapy in MM patients with low Hb is poor, and Hb is a factor affecting the efficacy of CAR-T therapy.

The Efficacy and Safety of Daratumumab-Based Regimen in Treatment of Multiple Myeloma Patients with Renal Impairment / 中国实验血液学杂志

OBJECTIVE@#To investigate the efficacy and safety of daratumumab in treatment of multiple myeloma (MM) patients with renal impairment (RI).@*METHODS@#The clinical data of 15 MM patients with RI who received daratumumab-based regimen from January 2021 to March 2022 in three centers were retrospectively analyzed. Patients were treated with daratumumab or daratumumab combined with dexamethasone or daratumumab combined with bortezomib and dexamethasone and the curative effect and survival were analyzed.@*RESULTS@#The median age of 15 patients was 64 (ranged 54-82) years old. Six patients were IgG-MM, 2 were IgA-MM,1 was IgD-MM and 6 were light chain MM. Median estinated glomerular filtration rate (eGFR) was 22.48 ml/(min·1.73 M2). Overall response rate of 11 patients with MM was 91% (≥MR), including 1 case of stringent complete response (sCR), 2 cases of very good partial response (VGPR), 3 cases of partial response (PR) and 4 cases of minor response (MR). The rate of renal response was 60%(9/15), including 4 cases of complete response (CR), 1 case of PR and 4 cases of MR. A median time of optimal renal response was 21 (ranged 7-56) days. With a median follow-up of 3 months, the median progression-free survival and overall survival of all patients were not reached. After treatment with daratumumab-based regimen, grade 1-2 neutropenia was the most common hematological adverse reaction. Non-hematological adverse reactions were mainly infusion-related adverse reactions and infections.@*CONCLUSION@#Daratumumab-based regimens have good short-term efficacy and safety in the treatment of multiple myeloma patients with renal impairment.

The In Vivo Intervention and Relative Mechanism of Genistein on the Inflammation and Thrombophilia in Lymphoma-Bearing Mice / 中国实验血液学杂志

OBJECTIVE@#To investigate the in vivo intervention and relative mechanism of Genistein (GEN) on tumor-associated inflammatory and tumor thrombophilia in lymphoma-bearing mice.@*METHODS@#Forty female Balb/c mice aged 5-6 weeks were injected with murine-derived Pro B-cell lymphoma cell line 38B9 to establish a lymphoma mouse model, which was randomly divided into control group, tumor-bearing group, GEN drug intervention group and cyclophosphamide (CTX）drug intervention group. Histopathologic was used to evaluate the tumorigenesis. Tumor formation was observed, and tumor tissues were collected of HE and immunohistochemical staining. ELISA and flow cytometry were used to detect the expression of inflammatory factors and the changes of thrombus indices in plasma after intervention of GEN and Cyclophosphamide (CTX) respectively. Immunohistochemistry method was used to detect the expression of CD19 in tomor tissues of tummor bearing mice.@*RESULTS@#After 14 days of tumor bearing, the mice were tumorigenic. The lymphoma cells were diffusely distributed in the tumor tissue and the expression of CD19 in the tumor tissue was positive. The inflammatory factors such as IL-6, NETs and CLEC-2, and thrombotic indices such as TF, FIB and D-D in lymphoma-bearing mice were significantly higher than those before tumor-injection and lower than those after drug-intervention (all P<0.05). The levels of CLEC-2 and D-D in GEN group were significantly lower than those in CTX group (P<0.05).@*CONCLUSION@#Tumor-associated inflammation and thrombophilia exist in lymphoma-bearing mice. GEN shows better anti-inflammatory and anti-thrombotic effects compared with CTX by interfering with tumor inflammatory factors.

Influence of the Ratio of Peripheral Hemoglobin-to-Red Cell Distribution width on the Prognosis of Patients with Diffuse Large B-cell Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To investigate the influence of peripheral hemoglobin (Hb)-to-red cell distribution width (RDW) ratio (HRR) on the prognosis of patients with diffuse large B-cell lymphoma (DLBCL).@*METHODS@#Data of 265 patients with diffuse large B-cell lymphoma (DLBCL) at the Affiliated Hospital of Xuzhou Medical University from January 2014 to December 2019 were retrospectively analyzed. 132 healthy people in the same period were used as normal control group. The best cut-off points of HRR was determined by receiver operating characteristics (ROC) curve; the chi-square test was used to analyze the correlation of clinical characteristics with HRR; the Kaplan-Meier method was used to compare the overall survival (OS) and progression-free survival (PFS) of HRR patients in different groups; the Cox proportional risk model was used for univariate and multivariate analysis.@*RESULTS@#The best cut-off value of HRR was 0.936, which was divided into low HRR group and high HRR group. The low HRR group had a higher ECOG score, higher incidence of advanced Ann Arbor stage, higher NCCN-IPI score, and elevated LDH level. K-M survival analysis showed that OS (P<0.001) and PFS (P<0.001) in the low HRR group were significantly shorter than that in the higher HRR group. The multivariate analysis revealed that HRR was an independent predictor of OS（HR＝0.379，95％CI：0.237-0.605，P<0.001） and PFS （HR＝0.384，95％CI：0.241－0.614，P<0.001） in DLBCL patients.@*CONCLUSION@#Low HRR（<0.936） in patients with DLBCL indicates a poor prognosis, which is an independent prognosis risk factor.

Analysis of Related Early Warning Indexes of Cytokine Release Syndrome in Multiple Myeloma Patients after CAR-T Treatment / 中国实验血液学杂志

OBJECTIVE@#To investigate the relationship between the levels of ferritin, C-reactive protein (CRP), lactate dehydrogenase (LDH) and interleukin-6 (IL-6) in peripheral serum and cytokine release syndrome (CRS) in patients with relapse and/or refractory multiple myeloma (R/R MM) after receiving chimeric antigen receptor T cells (CAR-T) immunotherapy.@*METHODS@#Twenty-eight patients with R/R MM were treated with 1×10@*RESULTS@#Among the 28 patients, 27 cases (96.4%) developed CRS, 24 cases (85.7%) in 1-2 grade CRS and 3 cases (10.7%) in 3-5 grade. The severity grade of CRS of 27 patients was positively correlated with the peak values of ferritin, CRP, LDH, and IL-6 in peripheral blood (r@*CONCLUSION@#After receiving CAR-T cellular immunotherapy, the incidence of CRS in patients with R/R MM is higher, but most of them are in grade 1 or 2. The severity of CRS is positively correlated with the levels of ferritin, CRP, LDH and IL-6 in peripheral blood.

Kinetics of Infiltrating CD3 / 中国实验血液学杂志

OBJECTIVE@#To explore the kinetics of infiltrated T cell in murine acute graft-versus-host disease (aGVHD) target organs after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and its relationship with tissue pathological damage and aGVHD progress.@*METHODS@#Male C57BL/6 (H-2K@*RESULTS@#Compared with BMT group, the number of infiltrated T cells in aGVHD target organs including liver, lung and gut increased since day 7 in BMT+T group (P<0.05). On day 14, 28, 40 and 47 after transplantation, more infiltrated CD3@*CONCLUSION@#Pathological damage of aGVHD target organs is induced by CD3

Expression and Significance of PD-1, TIM-3 and VISTA on T Cell of Acute Myeloid Leukemia Patients / 中国实验血液学杂志

OBJECTIVE@#To study the expression of multiple negative costimulatory molecules on peripheral blood T cells in patients with acute myeloid leukemia (AML) and its affection on prognosis.@*METHODS@#The peripheral blood samples from patients with newly diagnosed AML, complete remission (CR), and no-remission (NR) were collected, the expression levels PD-1、VISTA and TIM-3 in CD4 and CD8 T cells were detected by flow cytometry , and the clinical data of patients were analyzed.@*RESULTS@#The expression levels of PD-1、VISTA and TIM-3 of CD4 and CD8 T cells in the newly diagnosed AML patients were significantly higher than those in control group (P＜0.05). The expression levels of PD-1、TIM-3 and VISTA of CD4 and CD8 T cells in the CR group were significantly lower than those in newly diagnosed and the NR group (P＜0.05). The TIM-3 expression level positively correlated with VISTA expression level of CD4 and CD8 T cells in newly diagnosed AML patients (r=0.85 and 0.73). The VISTA and PD-1 expression level of CD4 T cells in newly diagnosed AML, NR after first induction chemotherapy and high risk patients significantly increased (P＜0.05), the TIM-3 expression level of CD8 T cells in high risk group significantly increased (P＜0.05), and the VISTA expression level of CD8 T cells in CBFβ-MYH11 mutation-positive group significantly decreased (P＜0.05).@*CONCLUSION@#The expression of PD-1、TIM-3 and VISTA in AML peripheral blood T cells may be involved in the immune escape of AML and can be the targets of treatment for acute myeloid leukemia patients.

Expression and Clinical Significance of EZH2 in Patients with Diffuse Large B Cell Lymphoma Accompanied by HBV Infection / 中国实验血液学杂志

OBJECTIVE@#To explore the expression and clinical significance of EZH2 in DLBCL patients accompanied by HBV infection.@*METHODS@#The clinicopathological data of 59 patients with DLBCL accompanied by HBV infection in our hospital from February 2015 to October 2017 were analyzed retrospectively. The patients were divided into HBV negative and HBV positive groups by serological testing before surgery. The expression of EZH2 was detected by immumohistochemical staining, and the clinicopathological characteristics and survival were analyzed and compared between these two groups.@*RESULTS@#There were 30 patients (50.8%) in the HBV negative group and 29 patients (49.2%)in the HBV positive group. The differences of age, LDH level and IPI score between two groups were statistically significant (P＜0.05). The expression of EZH2 in HBV- positive group was significantly higher than that in the HBV- negative group (P＜0.05), where the expression of EZH2 correlated with the expression of the BCL-6 (r=0.282, P＜0.05), especially in the GCB-DLBCL (r=0.549, P＜0.05). PFS was not significantly different between two groups of HBV (P＞0.05), while the PFS in the R-CHOP regimen group was higher than that in the CHOP regimen group (P＜0.05). COX multivariate analysis showed that both the chemotherapy regimen without R and the increased level of LDH were the risk factors affecting the prognosis of DLBCL patients (P＜0.05).@*CONCLUSION@#EZH2 highly expresses in HBV positive group, suggesting that the significance of EZH2 in DLBCL with HBV infection is worth further explore.

Effects of Combined Infusion of Mesenchymal Stem Cells and Endothelial Progenitor Cells on Lung Injury after Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

OBJECTIVE@#To investigate the effects of combined infusion of mesenchymal stem cells (MSC) and endothelial progenitor cells (EPC) on lung injury after hematopoietic stem cell transplantation (HSCT).@*METHODS@#The experiment was divided into normal control group, irradiation group, bone marrow cell transplantation group (BMT group), BMT+EPC group, BMT+MSC group and BMT+EPC+MSC group. The model of HSCT was established, on the 30th day after transplantation, the mice were sacrificed. Then lung tissue was taken for testing. The mRNA expression levels of VEGF, IL-18, IL-12b were detected by RT-PCR, and protein expression level of NLRP3 was detected by Western blot. The expression of MPO and CD146 was observed by immunohistochemistry assay.@*RESULTS@#The expression level of VEGF gene in BMT+EPC+MSC group was significantly higher than that in other groups (P＜0.01). The expression level of IL-18 and IL-12b gene was the highest in BMT group and the lowest in BMT+EPC+MSC group, and the difference was statistically significant (P＜0.05). HSCT could increase the expression of NLRP3 protein, and the BMT+EPC+MSC could significantly reduce the level of NLRP3 protein in lung cells, tending to normal. Compared with normal tissues, the BMT+EPC+MSC could improve the lung tissue structure more effectively, the expression of MPO positive cells was lower, and the expression of VEGF positive cells was higher.@*CONCLUSIONS@#The combined infusion of MSC and EPC can promote capillary regeneration, alleviate inflammation and promote lung repair after HSCT, which is superior to single EPC or MSC infusion.

Expression of Versican and Its Related Molecules in Patient with Multiple Myeloma and Its Clinical Significance / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression and clinical significant of VCAN and its related molecules in patients with MM.@*METHODS@#Ficoll density gradient centrifugation method was used to speared the bone marrow mononuclear cell in 25 cases of MM before and after treatment, the relative mRNA expression of VCAN and their related molecules (FAK, FN, MK, and HAS) in bone marrow was detected by real-time quantitative PCR, and their protein expression was determined by Western bolt.@*RESULTS@#The expression of VCAN, FK and FN in the effective group after treatment was significantly lower than that before treatment (P＜0.05), however, the expression of MK and HAS showed no statistically significantly different before and after treatment (P＜0.05). The expression of VCAN of patients in non remission group was significantly higher than that in control group (P＜0.05). The expression of FAK and FN of patients in no remission group was significant increased as compared with the patients in newly diagnosed group (P＜0.05). The relative expression of VCAN mRNA in the patients at 3rd stage was significantly higher than those at the 1st stage (P＜0.05) and control group but showed no significant difference to the patients at 2nd stage (P＜0.05). The expression of VCAN and its related proteins (FAK, MK, FN) showed positively correlation in bone marrow mononuclear cells of MM patients (P＜0.05). The correlation between VCAN and HAS was not statistically significant (r=0.259，P＞0.05). Survival analysis showed that the relative expression of VCAN mRNA was associated with OS (P=0.049) and PFS (P=0.041) in MM patients.@*CONCLUSION@#VCAN and its related molecules are highly expressed in MM patients; VCAN may act as potential biomarker in the development of multiple myeloma.

Clinical Significance of Platelet Closure Time in Patients with Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To explore the clinical significance of platelet closure time (PCT) in patients with multiple myeloma (MM).@*METHODS@#Peripheral blood samples of 50 newly diagnosed MM patients treated in our hospital from July 2018 to November 2019 and 34 healthy persons underuent physical at the same time were collected. PCT induced by collagen/epinephrine (CEPI) and collagen/adenosinediphosphate (CADP) in peripheral blood were detected by PFA-200，and the clinical data included age, sex, leukocyte count, hemoglobin level, platelet count and level of serum creatinine, cystatin c, blood calcium, β-microglobulin (β-MG), bone marrow plasma cells, light chain protein, as well as the MM types, ISS stage of patients were collected.@*RESULTS@#The level of PCT in MM patients was significantly higher than that in healthy persons; the level of PCT were significantly increased with the increasing of ISS stage in newly diagnosed MM patients; After chemotherapy with bortezomib/dexamethasone (BD), the level of PCT in 15 patients who were responded to the treatment was significantly lower than those before treatment.@*CONCLUSION@#The platelet closure time is abnormal in MM patients, moreover, relates to the progress of the disease. It has an important clinical significance for the evaluation of diagnostic stage and therapeutic efficacy evaluation of MM patients.

Th17 Cell Subset levels in Peripheral Blood of Patients with Aplastic Anemia and Their Clinical Significance / 中国实验血液学杂志

OBJECTIVE@#To study the level and clinical value of Th17 cell subset in the peripheral blood of the patients with aplastic anemia (AA).@*METHODS@#Eighty-five patients with aplastic anemia in our hospital from May 2017 to February 2018 were selected. Among them, 51 patients with poor curative effect were enrolled in group A. and the 34 patients with good curative effect were enrolled in group B, the 35 healthy persons were selected as controls. The levels of serum IL-17, IL-6, IL-4 and IFN-β were analyzed by ELISA; the levels of peripheral blood Th17 cell subset were analyzed by flow cytometry; the expression levels of RORγt and STAT3 mRNA in lymphocytes were analyzed by RT-PCR; the expression levels of RORγt and STAT3 protein in lymphocytes were analyzed by Western blot.@*RESULTS@#There was no significant difference in sex, age, WBC count and complications between group A and group B (P＞0.05). Non-severe aplastic anemia (NSAA) in group B accounted for 23 cases, and the NSAA ratio (23/34) was significantly higher than that in group A (20/51) (P＜0.05). The Hb level (86.25±7.9 g/L) and Plt count (54.7 ± 6.3×10/L) in group B were significantly higher than those in group A (P＜0.05). ELISA showed that the levels of IL-17 and IL-6 in group A were significantly higher than those in control group (P＜0.05); the levels of IL-17 and IL-6 in group B were significantly lower than those in group A (P＜0.05); the levels of IL-4 and IFN-β in group A were significantly lower than those in control group (P＜0.05); the levels of IL-4 and IFN-βin group B were significantly higher than those in group A (P＜0.05). Flow cytometry showed that the proportion of Th17 cells in peripheral blood of group A was higher than that of group B (P＜0.05). RT-PCR showed that the levels of RORγt and STAT3 mRNA in group A were significantly higher than those in control group (P＜0.05), and the mRNA levels of RORγt and STAT3 in group B were significantly lower than those in group A (P＜0.05). Western blot showed that the protein levels of RORγt and STAT3 in group A were significantly higher than those in control group (P＜0.05), and the protein levels of RORγt and STAT3 in group B were significantly lower than those in group A (P＜0.05).@*CONCLUSION@#The Th17 cells in peripheral blood of AA patients increase, and the inflammatory reaction in the patients are enhanced. It may be related with the rise of RORγt and STAT3 gene expression, which provides a research direction for clinical treatment of AA.

Relationship between Thrombotic Events and Peripheral Neutrophil-to-lymphocyte Ratio in Patients with Newly Diagnosed Essential Thrombocythemia / 中国实验血液学杂志

OBJECTIVE@#To explore the role of neutrophil-to-lymphocyte ratio（NLR) in patients with newly diagnosed essential thrombocythemia（ET) and its relationship with thrombotic events.@*METHODS@#The clinical and follow-up data of 150 ET patients were retrospectively analyzed. The risk factors of thrombotic events and role of NLR by statistical methods.@*RESULTS@#Age (P<0.01) and JAK2V617F mutation (P<0.01) were the independent risk factors for thrombotic events at diagnosis; WBC count (P<0.05), NLR (P<0.01), age (P<0.05) and thrombosis history at diagnosis (P<0.05) were independent risk factors for future thrombotic events. The ROC curve showed that NLR for prediction of future thrombotic events was suprior to other risk factors. The Kaplan-Meier analysis showed that the progress-free survival time in thrombotic events patients with higher NLR (median survival 22.3 months, 95% CI:17.8-26.8) was significantly shorter than that of patients with lower NLR (median survival 55.5 months, 95% CI:53.4-57.5) . After follow-up 60 months, the thrombosis progress-free survival in lower NLR patients reached 97.4%, while that in the patients with higher NLR was rate 46.7%.@*CONCLUSION@#NLR at diagnosis is a better predictive parameter for future thrombotic events than other clinical parameters in ET patients, but without corralations with thrombosis at diagnosis.

Survey and Establishment of the Hematological Parameter Reference Intervals for Adult in Xuzhou Area of China / 中国实验血液学杂志

OBJECTIVE@#To establish the reference intervals of the hematological parameters in normal adult people of Xuzhou erea.@*METHODS@#The red blood cells (RBC), platelets, white blood cells (WBC) and related parameters were detected by hematoanalyzers in 82514 healthy people including 41257 males and 41257 females in the Medical Center of the Affiliated Hospital of Xuzhou Medical University.@*RESULTS@#The range of RBC count: (4.33-5.51) ×10/L in male and（3.82-4.85）×10/L in female, the range of Hb level: (132-172) g/L in male and（107-145）g/L in female, the range of HCT: 40 %-50 % in male and 34 %-44 % in female, the range of platelet count: (113-268) ×10/L in male and (126-289) ×10/L in female, the range of WBC count: (4.00-9.40) ×10/L in male and （3.54-9.30）×10/L in female, the range of NEUT count: (1.91-5.76) ×10/L in male and（1.67-5.30）×10/L in female, the range of MONO count: (0.18-0.58) ×10/L in male and（0.16-0.52）×10/L in female, the range of LYM is（1.3-3.4）×10/L in male and（1.2-3.1）×10/L in female, etc.Conclusion: There is significant difference in the blood cell parameters between Xuzhou and other areas. Among them, the lower limit of Hb reference interval for adult women in Xuzhou area is obviously lower, and the upper and lower limits of adult Plt reference interval are significantly lower than other areas in China and abroad. The upper and lower limits of the WBC reference interval are close to the domestic areas, but lower than that in some foreign regions. Through this survey, the reference intervals of different hematologic parameters of healthy people in Xuzhou area primarily has been established, Some of the indexes, such as RBC, Hb, HCT, and Plt have significant sex differences. The reference intervals for them have been estublisted respectively.

Prognostic Significance of CD45CD117 Cells in Patients with Acute Myeloid Leukemia after Complete Remission / 中国实验血液学杂志

OBJECTIVE@#To investigate the predictive value of CD45CD117 phenotype-abnormal cells (hereinafter referred to as "abnormal cells") for relapse and prognosis in adult patients with acute myeloid leukemia (AML) within 2 weeks after the first complete remission (CR1).@*METHODS@#The clinical data of patients with newly diagnosed AML (non-acute promyelocytic leukemia) admitted in our department from July 1, 2014 to June 30, 2017 were analyzed retrospectively, and the relationship between clinical features at the initial diagnosis and the abnormal phenotype cells of CD45CD117 within 2 weeks after CR1 with the prognosis were analyzed.@*RESULTS@#A total of 91 patients with CD45CD117 abnormal cells were detected. The median age was 51 years old, the median WBC count was 11.60×109/L, and the median ratio of bone marrow blast cells was 0.35 at initial diagnosis. According to the FAB classification, 1 (1.1%), 7 (7.7%), 38 (41.7%), 20 (22.0%), 21 (23.1%) and 4 (4.4%) patients were classifice as M0, M1, M2, M4, M5, and M6, respectively. According to the NCCN risk stratification, 30 (33.0%), 51 (56.0%), and 10 (11.0%) patients were determined as good, moderate, and poor prognosis, respectively. The median ratio of abnormal cells within 2 weeks after CR1 was 1.8500 (0.0236-8.0000)%. The median time from initiation of induction therapy to the acquisition of CR was 46 days, median recurrence-free survival time was 319 days, and median overall survival time was 352 days. A total of 45 patients relapsed, of which 14 died; 46 patients did not relapse, of which 3 died. The cutoff of abnormal cells by receiver operating characteristic curve (ROC) analysis was 2.055% (Se=0.733，Sp=0.761). The abnormal cell ratio was＞2.055% in 44 patients, the median ratio of abnormal cells was 3.075%, among which 33 patients relapsed and 12 patients died; the abnormal cell ratio was ＜2.055% in 47 patients, the median ratio of abnormal cells was 1.150%, 12 patients relapsed and 5 patients died. Regression analysis showed that WBC count＞50×10/L and abnormal cell ratio＞2.055% were independent risk factors for recurrence. The abnormal cell ratio＞2.055% group had a 2-year RFS rate of 54.3% and a 2-year OS rate of 52.8%. The abnormal cell ratio＜2.055% group had a 2-year RFS rate of 86.6% (P=0.018), and a 2-year OS rate of 85.3% (P＜0.05).@*CONCLUSION@#For adult AML patients, CD45CD117 phenotypical abnormal cells ratio＞2.055% within 2 weeks after CR1 is an independent risk factor for recurrence, which also is an dverse factor for RFS and OS.

Effect of Stably Down-regulating FMI Expression of K562 Cells on Sensitivity of K562 cells to Imatinib Mesylate / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of stably down-regulating the FMI expression of K562 cells on the sensitivity of K562 cells to Imatinib (IM) and its possible mechanism.@*METHODS@#Western-blot was used to detect the expression of FMI protein in K562 cells and peripheral blood mononuclear cells from the patients with chronic myelogenous leukemia, chronic myeloid blast crisis and healthy volunteers. The specific interference sequences targeting at the human FMI gene were designed and ligated into the lentiviral vector LV3; the three plasmid system-packaged lentivirus particles were used to transfect K562 cells to screen K562 cells that stably down-regulated FMI. CCK-8 assay and flow cytometry were used to determine effect of IM on cell proliferation and apoptosis. The transcription level of FMI and Fz8 in leukemia cells was detected by fluorescent quantitative PCR. The protein expression levels of FMI, Fz8, NFAT1, BCR-ABL and β-catenin in leukemia cells were detected by Western-blot.@*RESULTS@#The expression of FMI protein could be detected in peripheral blood mononuclear cells of the patients with CML-BC and K562 cells, the FMI expression could not be detected in all the patients with CML-CP and healthy volunteers. The recombinant lentiviral vector LV3/FMI had been successfully constructed the lentivirus was packaged, and the K562 cells stably down-regulating the FMI protein were screened. After stable down-regulation of FMI expression in K562 cells, the proliferation rate of leukemia cells decreased and the apoptosis rate was increased under the same drug concentration. Both the transcription and protein expression levels of Fz8 decreased. The NFAT1 total protein level increased, as well as the nuclear translocation of protein was enhanced. There was no significant change in the expression level of BCR-ABL fusion protein. The expression level of β-catenin protein decreased.@*CONCLUSION@#After the stable down-regulation of FMI expression, the sensitivity of K562 cells to IM and apoptosis of cells increase, which are performed possibly by inhibiting the FMI-Fz8 signaling pathway and activating the Ca-NFAT and Wnt/β-catenin signaling pathway.

Role of Ca-NFAT Signaling Pathway in Ph ALL Drug-resistance Mediated by Bone Marrow Stromal Cells / 中国实验血液学杂志

OBJECTIVE@#To explore the role of Ca-NFAT signaling pathway in Ph-ALL drug resistance mediated by bone marrow stromal cells.@*METHODS@#The transcription level of NFAT mRNA in Sup-B15 cells and Ph ALL primary cells was detected by polymerase chain reaction. The expression of P-glycoprotein in Sup-B15 cells was detected by flow cytometry. The change of NFAT protein in Sup-B15 cells was detected by Western blot. AnnexinV/7-AAD was used to label cells. Flow cytometry was used to detect cell apoptosis; Fluo 3-AM dye was used to label cells, and flow cytometry used to detect changes of Ca concentration in leukemia cells.@*RESULTS@#NFAT expression could be detected in both Sup-B15 and Ph ALL primary cells; P-glycoprotein could not be detected by flow cytometry; CAS could significantly inhibit NFAT protein expression in clinically applied drug concentrations (2.5, 5 μmol/L); Clinically applied concentration of CAS (2.5, 5 μmol / L) has no significant effect on the apoptosis of Sup-B15 cells, while higher concentration of CAS (10 μmol / L) could induce apoptosis of Sup-B15 cells. Bone marrow stromal cells OP9 could, decrease the sensitivity of Sup-B15 cells and Ph ALL primary cells to imatinib (IM); After co-culture with bone were marrow stromal cells, the Ca concentration in Sup-B15 cells was enhanced, the levels of NFAT protein and nullear protein in sup-B15 cells also were enhanced. The addition of CAS in co-culture system could inlibit the Ca-NFAT signaling pathway, reduce the protective effect of OP9 on Sup-B15 cells.Conclution:The Ca-NFAT sigualing pathway, contributes to the survival of Ph ALL cells. Bone marrow stromal cells can mediate the resistance of Ph ALL cells to IM by activating Ca-NFAT signaling pathway.

Multivariate Analysis of Factors Influencing Recovery from Hemorrhagic Cystitis after Allo-HSCT / 中国实验血液学杂志

OBJECTIVE@#To analyze the incidence of hemorrhagic cystitis (HC) after allogeneic hematopoietic stem cell transplantation and the factors affecting HC, so as to provide clinical evidence for further treatment of HC.@*METHODS@#The HC of 113 patients after allogeneic hematopoietic stem cell transplantation in Affiliated Hospital of Xuzhou Medical University between the years 2014-2016 was analyzed respectively. All cases of HC were divided into HC group and non-HC(control) group. The follow-up time: from preeonditionig day to 180 d after transplantation. The 10 clinical parameters were selected for univariate analysis with COX regression analysis: sex, age (＜25 years and 25 years), primary disease, conditioning regimen with anti-thymoglobulin(ATG), sex-mismatch in recipients, haploidential HSCT, cytomegalovirus (CMV) viremia, EB viremia, graft-versus-host disease (GVHD), and primary disease relapse, the factors significant at the 0.1 level in univariate analysis should be further evaluated by multivariate analysis using a COX regression analysis. The difference was significant at P＜0.05 in multivariate analysis.@*RESULTS@#The HC occured in 31 of 113 patients (27.4%), with 5 cases of grade I (5.5%), 19 of grade II (16.8%), 5 of grade III (4.4%), and 2 of grade IV (1.8%). The median time of HC onset was 37 days (26-70 d) after transplantation. The median duration of HC was 14 days (5-55d). Univariate analysis showed that conditioning with anti-thymoglobulin (ATG) (RR=6.170, 95%CI: 1.875-20.306, P＜0.01), CMV viremia (RR=7.633, 95%CI：2.318-25.133) (P＜0.01), haploidentical HSCT (RR=0.307, 95%CI：0.137-0.686, P＜0.01), GVHD (RR=1.891, 95%CI：0.918-3.898, P＞0.05) were the risk factors for recovery from HC. The multivatiate analysis of above-mentioned risk factors with statistical significance showed that only CMV viremia (RR=4.770, 95%CI: 1.394-16.326, P＜0.05) was the indentified risk factor affecting the recovery from HC.@*CONCLUSION@#Monitoring CMV viremia and antivirotic treatment are effective measurs to prevent the occurrence of HC and promote the recovery from HC.

Significance of Peripheral Blood Lymphatic to Monocyte Ratio in the Progress of PGI-DLBCL / 中国实验血液学杂志

OBJECTIVE@#To explore the significance of lymphocyte to monocyte ratio (LMR) in the disease progress of primary gastrointestinal diffuse large B-cell lymphoma (PGI-DLBCL).@*METHODS@#The clinical data of 43 patients diagnosed as PGI-DLBCL in our hospital from January 2011 to December 2015 were collected, and the disease progress was followed up.@*RESULTS@#According to the ROC curve, the threshold value of LMR for 2 years PFS (%) of PGI-DLBCL patients was 2.6. Unvariate analysis showed that LMR (P＜0.05), large enclosed mass lesion (P＜0.01) and IPI (P＜0.05) were prognostic factors affecting PFS, the COX regression model multivariate analysis showed that LMR＜2.6 [ (risk ratio (RR)=3.083, 95%CI 1.828-8.313, P＜0.01], and large enclosed mass lesions (RR=2.718, 95%CI 1.339-6.424, P＜0.05) were the independent adverse prognostic factor for two years PFS.@*CONCLUSION@#Both LMR＜2.6 and large enclosed mass lesions relate with the progress of PGI-DLBCL.

Analysis of Cytogenetic Characteristics and Clinical Prognosis in 236 Patients with Myelodysplastic Syndrome / 中国实验血液学杂志

OBJECTIVE@#To investigate the relationship of cytogenetic features, clinical characteristics and prognosis in patients with myelodysplastic syndrome.@*METHODS@#The clinical characteristics and prognosis of 236 patients with MDS admitted to the Affiliated Hospital of Xuzhou Medical University from January 2013 to September 2017 were analyzed retrospectively, the follow-up observation and correlation analysis were performed.@*RESULTS@#There were 33 cases of refractory cytopenia with unilateral dysplasia (RCUD), 8 cases of refractory anemia with ring-shaped iron granulocytes (RARS), 70 cases of refractory cytopenia with multiple dysplasia (RCMD), 23 cases of refractory anemia (RA), 46 cases of refractory anemia with excessive blasts (RAEB-1), 48 cases of (RAEB-2), MDS-U 2 cases, simple del(5q) 6 cases. The detection analysis showed that the chromosome abnormality rate and complex chromosome abnormality rate in RAEB group (RAEB-1 + RAEB-2) and in non-RAEB group (RARS+RCMD+RCUD+RA) were 48.94% vs 43.94%, and 18.09% vs 12.69%, respectively, which were no statistically different. The grouping according to IPSS-R and IPSS showed that the chromosome abnormality rate gradually increased along with enhancement of risk stratifi-cation (P<0.05). The cytogenetic characteristics analysis showed that a total of 108 cases had chromosomal abnormalities, the detection rate was 45.76%, Out of 108 cases, 36 cases had complicated karyotypes, accounted for 15.25% of all patients. The types of chromosomal abnormalities mainly include numbers, structural abnormalities and mixed abnormalities. The chromosome abnormality with the highest detection rate was +8, accounted for 30.56% (33/100) of patients with chromosome abnormalities; followed by -7/7q-, del(5q), del(20q), etc. -7/7q-chromosome abnormalities accounted for 29.63% of all karyotypic abnormalities (including -7/7q-chromosome abnormalities alone and other chromosome abnormalities). The median age of the patients with MDS was 61 (13-88) years old, and the male-female ratio was 1.36∶1. Analysis of blood cell characteristics showed that the three lines were reduced or increased to varying degrees. The median WBC count was 2.8 (0.3-267.1)×10/L, the median Hb level was 69 (20-156) g/L, and the median Plt count was 55 (2-1733)×10/L. The 1 year OS rate in 32 cases of chromosome 7 abnormality and 128 cases of normal chromosome was 25% and 44.53%, respectively, the difference was statistically significant (χ=4.05, P<0.05) .@*CONCLUSION@#Chromosome karyotype is an independent factor affecting the prognosis of patients with MDS. It is important for the diagnosis, treatment and prognosis evalnation of patients with MDS. The overall prognosis of patients with abnormal chromosome 7 is poor. .