Positive MRD suggests a poor prognosis for ALL patients with or above CR2 before allogeneic transplantation / 中华检验医学杂志

Objective:To investigate the value of minimal residual disease (MRD) in prediction of prognosis in acute lymphoblastic leukemia (ALL) patients with or above complete remission 2 (CR2) underwent.Methods:A retrospective analysis was performed on 201 ALL patients who received allogeneic stem cell transplantation (allo-SCT) and pretransplant disease status ≥CR2 in Peking University People′s Hospital from January 2009 to December 2018. MRD was measured by multi-parameter flow cytometry at 1 month before transplantation and 1 month, 2 months, 3 months, 4 months, 6 months, 9 months or 12 months after transplantation. To investigate the influence of dynamic changes of MRD before and after transplantation on prognosis.Results:201 ALL patients, including 126 males and 75 females, with a median age of 18 years. The 3-year cumulative incidence of relapse (CIR), non-relapse mortality (NRM), leukemia-free survival (LFS) and overall survival (OS) of all cases were 34%, 16%, 50%, and 56%, respectively. Positive pre-SCT MRD patients with higher 3-year CIR (47% vs 26%, P=0.003), lower 3-year LFS (40% vs 55%, P=0.047) and OS (42% vs 60%, P=0.065) than those with negative one. Subjects with positive post-MRD had higher 3-year CIR (73% vs 22%, P<0.001) and lower 3-year LFS (28% vs 56%, P=0.005) and OS (32% vs 60%, P=0.040) compared with those with negative one. Multivariate analysis showed that both pre-MRD and post-MRD were associated with higher CIR ( HR=1.823, P=0.018; HR=3.474, P<0.001), lower LFS ( HR=1.779, P=0.007; HR=2.185, P=0.001) and OS ( HR=1.609, P=0.034; HR=1.970, P=0.001). Negative pre-and post-SCT MRD group had lower 3-year CIR (17%, 42%, 82%; P<0.001) and higher 3-year LFS (61%, 44%, 18%; P<0.001) and OS (63%, 47%, 27%; P<0.001) compared with those unrisen post-SCT MRD group, and increased post-SCT MRD group. Multivariate analysis showed that pre-and post-SCT MRD dynamics were associated with CIR, LFS and OS ( P<0.01 for all) independently. The pre-and post-SCT MRD dynamics could better distinguish CIR (C=0.669) from that of pre-SCT MRD (C=0.587) and post-SCT MRD (C=0.629). Conclusion:Our data suggest that pre-SCT MRD, post-SCT MRD and the dynamic peri-SCT MRD could be used to predict transplant outcome of ALLpatients with or above CR2 who underwent allo-SCT.

Characteristic and prognostic significance of leukemia stem cells associated antigens expressions in t (8;21) acute myeloid leukemia / 中华血液学杂志

Objective@#To investigate the characteristic and prognostic significance of leukemia stem cells associated antigens expressions including CD34, CD38, CD123, CD96 and TIM-3 in t (8;21) AML.@*Methods@#Bone marrow samples of 47 t (8;21) AML patients were collected at diagnosis from October 2015 to April 2018 in Peking University Peoples’ Hospital, then flow cytometry method was performed to detect the expression frequencies of CD34, CD38, CD123, CD96 and TIM-3 to analyze the relationship between leukemia stem cells associated antigens expressions and relapse.@*Results@#Of 47 t (8;21) AML patients tested, the median percentages of CD34+CD38-, CD34+ CD38-CD123+, CD34+CD38- CD96+ and CD34+ CD38- TIM-3+ cells among nucleated cells were 2.37%, 0.24%, 0.27% and 0.06%, respectively. All the frequencies of CD34+CD38-, CD34+CD38-CD123+, CD34+CD38-CD96+ and CD34+ CD38-TIM-3+ cells had no impact on the achievement of CR after the first course of induction. All higher frequencies of CD34+CD38-, CD34+CD38-CD123+, CD34+CD38-CD96+ cells were related to higher 2-year CIR rate. Whereas, the frequency of CD34+ CD38- TIM-3+ cells had no impact on CIR rate. Both high frequency of CD34+ CD38- cells and the high level of minimal residual diseases (patients with <3-log reduction in the RUNX1-RUNX1T1 transcript level after the second consolidation therapy) were independent poor prognostic factors of CIR[P=0.025, HR=6.9 (95%CI 1.3-37.4) ; P=0.031, HR=11.1 (95%CI 1.2-99.2) ].@*Conclusion@#Different leukemia stem cells associated antigens had distinct prognostic significance in t (8;21) AML. High frequencies of CD34+ CD38-, CD34+ CD38- CD123+ and CD34+CD38-CD96+ cells at diagnosis predicted relapse in patients with t (8;21) AML.

A comparison between cDNA and DNA samples for the detection of c-KIT exon 17 mutation in acute myeloid leukemia patients / 中华检验医学杂志

Objective To investigate the impact of sample typeon the detection of c-KIT exon 17 mutation in acute myeloid leukemia (AML) patients. Methods A retrospective study was conducted on 51 bone marrow samples collected from 37 AML patients [17 maleand 20 female, with a median age of 33 (range from 1 to 82)] at diagnosis or after treatment from June 2016 to August 2018. Of the 37 cases of AML, 24 were t(8; 21) AML, 11 were inv(16)/t(16;16) AML and 2 were non-CBF-AML. RNA and DNA were simultaneously extracted from every sample. PCR followed by Sanger sequencing were used to screen c-KIT exon 17 mutation, and the comparisons were made between paired cDNA and DNAsamples. Results (1) Of the 51 paired samples, 14 pairs were simultaneously detected positive for c-KITmutation in both of cDNA and DNA samples, but 17 pairs were detected negative in both, and the remaining 20 pairswere only detected positive for the mutation in cDNA but not in DNA, with an inconsistency rate of 39.2％. The positive rate of detecting c-KITmutation was significantly higher in cDNA than in DNA samples (66.7％vs 27.5％,P=0.000073). (2)Inconsistent mutation results between paired cDNA and DNA samples occurred in t(8;21)AML, inv(16)AML and non-CBF-AML patients with the inconsistency rate of 36.4％(12/33), 27.2％(3/11) and 71.4％ (5/7), respectively. (3)The inconsistency rate was significantly higher in samples collected after treatment compared with those collected at diagnosis (72.7％vs 13.8％, P=0.00003). (4) All 5 serially monitored patients with c-KITmutation had inconsistency in mutation detection between cDNA and DNA samples during follow up. Conclusion cDNA improves the detection of c-KIT exon 17 mutation in AML patients compared with DNA, which is especially common after treatment.

Minimal residual disease in adults with Philadelphia chromosome negative acute lymphoblastic leukemia in high-risk / 中华血液学杂志

Objective@#To explore the significance of minimal residual disease (MRD) in predicting prognosis and guiding therapy of adults with Philadelphia-chromosome negative acute lymphoblastic leukemia (Ph- ALL) in high-risk.@*Methods@#Data of newly diagnosed adults with Ph- ALL in high-risk who achieved CR were reviewed. Variables associated with outcome were identified by COX regression model and Landmark analysis.@*Results@#A total of 177 patients, 99 (56%) cases male with a median age of 40 years (range, 16-65 years) were included in this study. Of them, 95 (54%) patients received allo-HSCT in CR1. Multivariate analyses showed that MRD negativity after the first cycle of consolidation (HR=0.52, 95%CI 0.30-0.89, P=0.017) and achieving CR within 4 weeks (HR=0.43, 95%CI 0.24-0.79, P=0.006) were the factors significantly-associated with longer DFS, and allo-HSCT was associated with both longer DFS (HR=0.13, 95%CI 0.08-0.22, P<0.001) and OS (HR=0.24, 95%CI 0.15-0.41, P<0.001) . Landmark analysis was performed on 121 patients, of 85 patients achieving MRD negativity after the first cycle of consolidation, multivariate analyses showed that MRD negativity after the third cycle of consolidation was significantly-associated with longer DFS (HR=0.18, 95%CI 0.05-0.64, P=0.008) and OS (HR=0.14, 95%CI 0.04-0.50, P=0.003) . For the patients achieving MRD negativity after both the first and the third cycles of consolidation, the 3-year DFS rate in the allo-HSCT cohort had a higher trend compared with that in the chemotherapy cohort (75.2% vs 51.3%, P=0.082) , however, the 3-year OS rates in the 2 cohorts were similar (72.7% vs 68.7%, P=0.992) . In those with MRD positivity after the first and/or the third cycle of consolidation, 3-year DFS (64.8% vs 33.3%, P=0.006) and OS (77.0% vs 33.3%, P=0.028) rates in the allo-HSCT cohort were significantly higher than those in the chemotherapy cohort, and similar to those in the cohort achieving MRD negativity after both the first and the third cycles of consolidation and receiving allo-HSCT.@*Conclusions@#MRD negativity after the first cycle of consolidation was a predictor for better outcome in adults with Ph- ALL in high-risk. The survival advantage of the allo-HSCT cohort was not pronounced compared with that in the chemotherapy cohort even in those with high-risk features but achieving MDR negativity after both the first and third cycles of consolidation. However, allo-HSCT could be a good option for the patients with MRD positivity after the first and/or the third cycle of consolidation.

Effects of hematological complete remission status before transplantation on outcomes of patients with B-cell acute lymphoblastic leukemia receiving haploidential stem cell transplantation / 临床检验杂志

Objective@#To investigate the association of hematological complete remssion (HCR) status on the outcomes of the patients with B-cell acute lymphoblastic leukemia (B-ALL) who were undergoing haploidentical stem cell transplantation (Haplo-SCT). @*Methods@#Retrospective analysis was performed on 317 patients with B-ALL who received Haplo-SCT with HCR before transplantation in the Institute of Hematology, Peking University from September 2012 to June 2016. A Cox proportional hazards model was used to analyze the effects of HCR status before transplantation on the outcomes of Haplo-SCT. @*Results@#The 3-year cumulative incidences of non-relapse mortality (NRM) and cumulative incidence of relapse (CIR) were 15% and 15%, respectively. The 3-year probabilities of leukemia-free survival (LFS) and overall survival (OS) were 71% and 74%, respectively. There was no statistical difference for 3-year NRM, CIR and LFS among the HCR patients with recovery of absolute neutrophil count (ANC) and platelet (CR) group, without recovery of ANC and with or without recovery of platelet (CRi) group and those in HCR with recovery of ANC but without recovery of platelet (CRp) group (P value ＞0.05 for all). The probability of OS in cases of CR group was significantly higher than that of CRi group (76% vs 59%，P=0.049). Multivariate analysis showed that factors associated with CIR included pre-transplantation minimal residual disease (P=0.006) and chronic GVHD (P=0.020). Platelet engraftment was associated with NRM, LFS, and OS (P＜0.001 for all). Grades Ⅲ-Ⅳ GVHD was associated with NRM (P＜0.001) and OS (P=0.035). Chronic GVHD was correlated with LFS (P＜0.001). @*Conclusion@#Our results indicate that no effect of HCR status before transplant on the outcomes was observed in patients with B-ALL who underwent Haplo-SCT.

Clinical implication of minimal residue disease monitoring by WT1 gene detection and flow cytometry in myelodysplastic syndrome with allogeneic stem cell transplantation / 中华血液学杂志

Objective@#To investigate the clinical significance of minimal residual disease (MRD) monitoring by using WT1 gene and flow cytometry (FCM) in patients with myelodysplastic syndrome (MDS) who receiving allogeneic stem cell transplantation (allo-HSCT).@*Methods@#WT1 gene and MDS-related abnormal immunophenotype were examined by real-time quantitative polymerase chain reaction (RQ-PCR) and FCM, respectively. The bone marrow samples were collected from patients with MDS who received allo-HSCT from Feb, 2011 to Oct, 2015 in Peking University People’s Hospital before and after transplantation.@*Results@#Among 92 MDS patients, 40 (48.2%) patients were positive for WT1 (WT1+) and 9 (10.8%) patients were positive for flow cytometry (FCM+). 27 patients (29.3%) met the criteria of our combinative standard, MRDco (MRDco+). Only FCM+ post-transplant (P<0.001) and MRDco+ (P=0.017) were associated with relapse. The cumulative incidence of relapse (CIR) at 2 years were 66.7% and 1.2% (P<0.001) in FCM+ and FCM- groups. MRDco+ group had a 2-year CIR of 23.0% while MRDco- group had a 2-year CIR of 1.6% (P=0.004). The specificity of post-transplant WT1, FCM and MRDco to predict relapse was 59.0%, 96.4% and 74.7%, respectively. The sensitivity of these three MRD parameters to predict relapse was 66.7%.@*Conclusion@#Post-transplant FCM and MRDco are useful tools to monitor MRD for MDS after transplantation. The preemptive intervention based on MRDco is able to reduce the relapse rate.

The comparison of predicting clinical outcomes between immunolophenotype and hematological complete remission before human leukocyte antigen-matched sibling donor transplantation in acute myeloid leukemia / 中华血液学杂志

Objective@#To assess the prognostic significance of immunophenotype complete remission (ICR) and hematological complete remission (HCR) before human-leukocyte antigen (HLA)-matched sibling donor transplantation (MSDT) in acute myeloid leukemia (AML) patients.@*Methods@#A cohort of 182 AML (non-APL) patients undergoing MSDT in HCR was retrospectively studied [including complete remission with ANC and PLT recovery (CR), CR with incomplete PLT recovery (CRp), CR with inconplete ANC and PLT recovery (CRi)]; ICR was determined as undetective minimal resudial disease (MRD) by multi-parameter flow cytometer.@*Results@#①Of the 182 patients, 97 were male, 85 female, and the median age was 41(4-62) years. ②The CR and CRi+CRp rates were 80.8% (147/182) and 19.2%(35/182), respectively; The 4-year cumulative incidence of relapse[CIR, (11.0±4.3)% vs (16.0±7.1)%, χ2=0.274, P=0.600], non-relapse mortality[NRM, (14.0±4.3)% vs (9.0±6.3)%, χ2=0.913, P=0.339], leukemia-free survival[LFS, (75.0±5.1)% vs (75.0±8.3)%, χ2=0.256, P=0.613], and overall survial [OS, (77.0±5.2)% vs (80.0±8.1)%, χ2=0.140, P=0.708] were comparable between the CRp+CRi and CR groups. ③Compared with the non-ICR group (n=35), the ICR group (n=147) showed lower 4-year CIR [(11.3±3.4) % vs (55.2±8.8) %, χ2=32.687, P<0.001], better 4-year LFS [(76.2±4.7)% vs (32.8±8.7)%, χ2=26.234, P<0.001] and OS[(79.0±4.7)% vs (39.0±9.1)%, χ2=25.253, P<0.001], and comparable NRM[(12.5±4.1)% vs (12.0±7.1)%, χ2=1.002, P=0.656]. ④Mulitvariate analysis confirmed the independent prognostic value of ICR in lower CIR [HR=11.026(95%CI 4.685-25.949), P<0.001], higher LFS [HR=5.785 (95% CI 2.974-11.254), P<0.001] and OS[HR=5.578 (95% CI 2.575-27.565), P<0.001].@*Conclusion@#The results indicated that ICR instead of HCR pre-transplantation had a significant prognostic value in AML patients undergoing MSDT.

Transformation from promyelocytic leukemia with t (15; 17) ( q22; q21) to acute monocytic leukemia with t (11; 17) (q23; q21) in a case / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To report on a case of therapy-related acute monocytic leukemia(t-AML) with t(11;17) (q23;q21)/MLL-AF17q after successful treatment for acute promyelocytic leukemia(APL) with t(15;17) (q22;q21)/PML-RARα.</p><p><b>METHODS</b>A MICM method (bone marrow morphology(M), immunophenotype(I), cytogenetics(C), and molecular biology(M)) was used for the diagnosis and classification of the disease at the time of onset and transformation.</p><p><b>RESULTS</b>The patient was initially identified with typical morphology and immunophenotype of APL. She has carried t(15;17)(q22;q21) and PML-RARα fusion gene but was without t(11;17)(q23;q21) or MLL gene abnormalities. After 13 months of successful treatment, she has transformed to AML with typical morphology and immunophenotype. t(11;17)(q23;q21) and MLL-AF17q fusion gene were detected in her bone marrow sample, while no PLZF-RARα fusion gene was detected by real-time quantitative reverse-transcription PCR(RQ-PCR) and fluorescence in situ hybridization(FISH).</p><p><b>CONCLUSION</b>t-AML is a serious complication after successful treatment of APL. t(11;17)(q23;q21) is not specific for the diagnosis of variant APL and can also be detected in t-AML. RQ-PCR and FISH are essential for the diagnosis of such patients.</p>

Comparison and analysis of minimal residual disease in multiple myeloma patients detected by 4-color and 8-color fluorescence antibody panels / 中华血液学杂志

Objective@#To explore and compare 4-color and 8-color fluorescence antibody panels for detecting minimal residual disease of multiple myeloma patients after therapy.@*Methods@#One 8-color antibody panel was built including CD38 and CD138 for the identification of plasma cells (PCs) , membrane antigen CD45, CD19, CD56 and CD117, cytoplasmic Kappa (cκ) and Lambda (cλ) light chain antigen. Six tubes of 4-color panels were built, among them, membrane antigen CD45/CD19, CD56/CD117, CD19/CD56 and light chains were analyzed combined by CD38/CD138 for PCs gate in the tubes M1-3 and tube C-κ/λ, respectively; CD19 or CD45 and cκ/cλ light chains were detected in the tube MC1-CD38 for CD38/SSC identified PCs gate and tube MC2-CD138 for CD138/SSC identified PCs gate separately. Twenty normal volunteer bone marrows and seventy-three specimens from multiple myeloma patients after complete remission were measured and analyzed.@*Results@#MRD positive samples were discriminated in 82.19% of the specimen evaluated through either abnormal plasma cells (aPCs) or clonal plasma cells (cPCs) by 8-color antibody panel. Among of them, consistency was 89.04%. The median percentage of cPCs was 0.105%. The lowest sensitivity of experiment was 0.004%. Percentage of PCs identified by CD38/ SSC gate was higher than that by CD38/CD138 (P<0.001) and CD138/SSC gate (P=0.001) apparently. The lowest MRD positive rate was found in tube C (65.75%) , which lower than 8-color panel obviously (P=0.024) . The percentages of aPCs measured by tube M2-56/117 were significantly lower than other tubes (P=0.014) . MRD positive rate determined by cPCs was higher than that by aPCs both in the tube MC1-CD38 and tube MC2-CD138, whose concordance rate were 68.49% and 79.45%, respectively. Compared with 8-color panel, tube MC2-CD138 the best choice among six tubes of 4-color panels, which has the best sensitivity, accuracy and negative predicted value, higher positive predicted value and specificity. Tube M1-45/19 was the second choice.@*Conclusions@#CD38/CD138 are the best markers for gating PCs. Membrane antigen and cκ/cλ detected simultaneously is a better method for MM MRD detection and 8-color antibody panel is an ideal approach. Two tubes of 4-color antibody combination, M1-45/19 and MC2-CD138 are recommended in the 4-color panels.

Bortezomib improves progression-free survival in multiple myeloma patients overexpressing preferentially expressed antigen of melanoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Significant efforts have been made to identify factors that differentiate patients treated with novel therapies, such as bortezomib in multiple myeloma (MM). The exact expression pattern and prognostic value of the cancer/testis antigen preferentially expressed antigen of melanoma (PRAME) in MM are unknown and were explored in this study.</p><p><b>METHODS</b>The transcript level of PRAME was detected in bone marrow specimens from 100 newly diagnosed MM patients using real-time quantitative polymerase chain reaction, and the prognostic value of PRAME was determined through retrospective survival analysis. PRAME expression higher than the upper limit of normal bone marrow was defined as PRAME overexpression or PRAME (+).</p><p><b>RESULTS</b>Sixty-two patients (62.0%) overexpressed PRAME. PRAME overexpression showed no prognostic significance to either overall survival (n = 100) or progression-free survival (PFS, n = 96, all P > 0.05) of patients. The patients were also categorized according to regimens with or without bortezomib. PRAME overexpression tended to be associated with a lower two-year PFS rate in patients treated with non-bortezomib-containing regimens (53.5% vs. 76.9%, P = 0.071). By contrast, it was not associated with the two-year PFS rate in patients with bortezomib-containing regimens (77.5% vs. 63.9%, P > 0.05). When the patients were categorized into PRAME (+) and PRAME (-) groups, treatment with bortezomibcontaining regimens predicted a higher two-year PFS rate in PRAME (+) patients (77.5% vs. 53.5%, P = 0.027) but showed no significant effect on two-year PFS rate in PRAME (-) patients (63.9% vs. 76.9%, P > 0.05).</p><p><b>CONCLUSION</b>PRAME overexpression might be an adverse prognostic factor of PFS in MM patients treated with non-bortezomib-containing regimens. Bortezomib improves PFS in patients overexpressing PRAME.</p>

Effects of different nutrition intervention on alimentary tract reconstruction in knowledge and practice nutrition and efficacies of patients with type 2 diabetes mellitus / 中华全科医师杂志

A total of 206 patients of type 2 diabetes mellitus (T2DM) were assigned into the trial group (n =156) and the control group (n =50) according to whether real object-based diabetic diet was provided.Traditional nutritional education for both groups during hospitalization.The patients in the trial group received a real object-based diabetic diet guided by a nutritionist who adjusted feeding regimens and skills before and after operation.The control group had a free access to a normal diet.The patient understanding of knowledge,attitude and practice (KAP) was examined by nutrition surveys.And their postoperative nutrition states and complications were assessed by questionnaires during a 6-month follow-up.The total average score of cognitive divided was 93.4 ± 7.5,the average score of diet therapy knowledge 13.8 ± 1.6 and the average scores of attitude and practice 9.6 ± 1.9 and 32.2 ±3.9.Significant differences existed between two groups (P ＜ 0.05).The incidence rates of hypoglycemia,hyperglycemia and gastrointestinal obstruction were lower than the control group (P ＜ 0.05,P ＜ 0.01).It suggested that real object-based diabetic diet guidance plus traditional nutrition education could improve the KAP level and may lower the incidence of postoperative complications in T2DM patients with gastric bypass.

Blastic plasmacytoid dendritic cell neoplasm: a case report / 中华皮肤科杂志

A 19-year-old female presented with a dark erythematous maculopapule measuring 2 cm in diameter on the left lower limb for more than one year.Physical examination revealed an enlarged cherry-like lymph node in the right inguinal region.Histopathology of the lesion revealed that the normal structure of skin disappeared absolutely,and there was a diffuse infiltrate of medium-sized lymphoid tumor cells with unclear nucleoli.Immunohistochemically,the tumor cells strongly expressed CD4,CD56 and CD123,and partially expressed terminal deoxynucleotidyl transferase (TdT).Flow cytometric analysis of bone marrow aspirates showed that abnormal cells amounted to 59.9％,and were positive for CD123st (strong),human leucocyte antigen-DRst (strong),CD56,CD304 (blood dendritic cell antigen-4),CD7,CD11b,CD33,CD36 and chemokine (C-X-C motif) receptor 4,CD13 (dim),CD4 (partial) and CD117 (partial).Transmission electron microscopy of bone marrow cells revealed that there were massive uniformly sized lymphoid cells with thick processes on cell surfaces,a high nucleus/cytoplasm ratio,centrally located round nuclei occasionally with deep gyrus-like notches,clumped marginated heterochromatin and multiple nucleoli.Moreover,a small amount of cytoplasm was observed in the lymphoid cells with the presence of mitochondria and endoplasmic reticulum arranged in concentric circles,which were characteristic of plasma cells.The features of both dendritic cells and plasma cells were found in the tumor cells through transmission electron microscopy.Based on the above findings,a diagnosis of blastic plasmacytoid dendritic cell neoplasm (BPDCN) was made.

Study on IFN-γ/TGF-β Level in a Model of Acute Inflammation and Melanoma / 中国肿瘤临床

Objective: To build a mouse tumor model with a manufactured surgical wound representing acute inflammation, and to evaluate the relationship between acute inflammation or wound healing and the process of tumor growth. Then to observe the impact of IFN-γ/TGF-β on tumor growth. Methods: Male C57BL mice of six weeks were used and divided into the experiment group and the control group. The B16F10 mela-noma cell suspension was injected into the left groin area of each mouse. A wound measured 1 cm in diame-ter was built on the opposite side of bodies in the experiment group when tumor volume was about 0.5 cm~3.The expression of IFN-γ/TGF-β in blood serum and tumor tissues were examined by ELISA. In order to fur-ther confirm the effect of TGF-β on tumor growth, another 16 mice models with melanoma were established and 8 of them received IFN-γ injection (the experiment group). Results: When acute inflammation had influenc-es on tumor, a two-phase development was presented. In the early phase, the growth of tumor in the mice with wound was slower than that in the control group. In the early phase, the release of IFN-γ was higher and the release of TGF-β was lower in the experiment group. In the later phase, the growth of tumor in the mice with wound was similar to that in the controls and the release of TGF-β was higher. In vivo experiment con- firmed the above results. In the early phase, the release of TGF-β was not significantly different between the experiment group and the control group (P>0.05). In the later phase, the release of TGF-β in the experiment group was higher than that in the control group (P<0.05). Conclusion: In the early phase of acute inflamma-tion, inhibitory effects of IFN-y on tumor growth were presented. In the later phase, the inhibited tumor was re-sistant to IFN-γ through the release of TGF-β to balance the effect of inflammatory factors on tumor cells.

The significance of dynamic detection of WT1 expression on patients of hematologic malignancy following allogeneic hematopoietic stem cell transplantation / 中华内科杂志

Objective To evaluate preliminarily the significance of dynamic detection of Wilms'tumor gene(WT1)expression level on monitoring minimal residual disease(MRD)and predicting clinical relapse in patients of malignancy following allogeneic hematopoietic stem cell transplantation(allo-HSCT).Methods WT1 expression level WaS measured with real.time quantitative reverse transcription polymerase chain reaction(RQ-RT-PCR)method on 102 bone marrow specimens from 31 patients following allo.HSCT.WT1 expression level was determined as the ratio of WT1 mRNA to ABL mRNA times 100%.Resuits The levels of WT1 expression showed significant difference between the relapsed group and the non-relapsed group(P＜0.01),with 0.80%and 0.17%as their median expression level respectively.After dynamic measuring WT1 expression level on patients in the relapsed group.the level was found to increase significantly as compared with those during or before the clinical hematological relapse.both being＞1.0%.The level at the time of relapse Was significantly higher than the latest previous one(P＜0.05).Conclusion Dynamic detection of WT1 expression level with RQ-RT-PCR may be of help in monitoring MRD and warning clinical relapse Oil the patients following allo-HSCT.

Effect of acupuncture of Baihui on neurotransmitter of post cerebral infarction depression / 中华中医药杂志

Objective:To observe neurotransmitter changes of post cerebral infarction depression (PCID) before and after being treated with amitriptyline or acupuncture of Baihui,in order to explore a good way with a less toxicity and good effect of treatment on PCID.Methods:The 60 patients were divided into treatment group and control group.Besides the basic treatment,the treatment group was given acupuncture of Baihui (DV20) and point injection therapy,the control group was given amitriptyline. Both before and after treatment,compared the two groups with HAMD score,neurologic impairment score,plasma 5-HT and NE,and observed the adverse events and side effects.Results:The results of two groups in monoamine neurotransmitter 5-HT(5-HT),norepinephrine (NE) and the HAMD scale were significantly improved.Conclusion:Acupunture of Baihui and point injection of Herba Erigerontis had equal effect to the amitriptyline on PCID.Furthermore,it had fewer venenosus side effects.It provided a new way for the use of the traditional Chinese medicine in treating PICD.

Inhibition of K562 cell growth and tumor angiogenesis in nude mice by antisense VEGF(121) cDNA transfection / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of antisense vascular endothelial growth factor (VEGF)(121) cDNA transfection on the growth of K562 cells in nude mice.</p><p><b>METHODS</b>K562 cells transfected with the antisense (AS) or sense (S) VEGF(121) cDNA, and the vector (V, pcDNA3) alone were transplanted subcutaneously into nude mice and the growth of the transfected cells in vivo was investigated. The effects of transfected K562 cells on human bone marrow endothelial cells (BMEC) were analyzed by MTT assay, the microvessel density (MVD) in tumor mass by vWF immunohistochemistry stain.</p><p><b>RESULTS</b>K562/V tumor grew more slowly [(207.5 +/- 192.9) mm(3) vs (445.0 +/- 150.9) mm(3), P < 0.05] and K562/S tumor more rapidly than K562/V tumor did [(1 174.6 +/- 508.7)/mm(3) vs (445.0 +/- 150.9) mm(3), P < 0.01]. K562/S cell culture supernatant was more strongly in promoting the proliferation of BMEC than K562/V supernatant did, but K562/AS supernatant resulted in a marked decrease of the promoting effect as compared with K562/V's. The MVDs in K562/AS, K562/S, and K562/V tumors were [(11.0 +/- 7.6)/0.72 mm(2) vs (50.8 +/- 11.7)/0.72 mm(2) vs (18.9 +/- 7.0)/0.72 mm(2)], respectively.</p><p><b>CONCLUSIONS</b>Antisense VEGF(121) cDNA transfected K562 cells show growth retardation in transplanted nude mice, decrease of tumor MVD, and decrease of promoting BMEC proliferation capacity.</p>

Analysis the immunophenotyping of chronic lymphoid leukemia / 中华检验医学杂志

Objective To investigate the characteristic immunophenotype of B cell chronic lymphoid leukemia in china. Method Single and multiparameter flow cytometry were used to analysis 163 cases of B cell chronic lymphoid leukemia. Results 71.8%(117/163) of cases co-expressed CD5 and B cell markers. The patients were classified into category of B cell chronic lymphocytic leukemia(B-CLL), hairy cell leukemia(HCL) and other B-cell lymphoproliferative disorders(LPDs) by using the scoring system that was recommended by world health organization (WHO). The B-CLL typically display the composite phenotypes: CD5+,CD23+,CD20+,CD19+,HLA-DR+,but the CD22,CD11c,CD25 and FMC7 were variable present in some B-CLL cases.CD103 seems the most specific marker for HCL.To differentiate diagnosis of atypical B-CLL with B-prolymphocytic leukemia(B-PLL) or mantle cell lymphoma(MCL), one must not rely exclusively on immunophenotypic dates, cytogenetic or molecular biology detection would be helpful. The index of froward scatter( FSC) and antigens expression of tumor B cells could be calculated by dividing the relevant value of residual normal T cell within same sample as internal control, so the cell size and the intensity of antigen expression could be comparable each other and quantitative between different investigations. Conclusion immunophenotypic analysis is an extremely useful adjunct in the diagnosis of chronic lymphoid leukemia.

Standardization and Quality Control in Flow Cytometric Enumeration of CD34(+) cells / 中国实验血液学杂志

The enumeration of CD34(+) cells by flow cytometry is commonly employed to assess hematopoietic stem/progenitor cells in cord blood, peripheral blood and apheresis products. Interlaboratory variation of CD34(+) cells enumeration is exceedingly large. Factors contributing to those variation, currently established flow cytometry protocols and comparison between these protocols were reviewed. CD34(+) cells in 45 cord blood samples, 12 normal bone marrows and 4 apheresis products were also enumerated in our laboratory by using ProCUNT kit, and results showed that the ProCUNT is highly standardizaed and could assist in reducing interlaboratory variation of CD34(+) cells.

The Function of Dendritic Cells Derived from Chronic Myeloid Leukemia / 中国实验血液学杂志

The purpose of this study was to investigate the function of dendritic cells derived from chronic myeloid leukemia (CML-DC). Mononuclear cells were prepared from bone marrow and peripheral blood of 24 patients with CML, and the DCs were obtained by incubation of MNCs with media containing GM-CSF, IL-4 and TNF-alpha. The phenotype of CML-DCs was identified by flow cytometry. FITC-dextran uptake, (3)H-TdR incorporation or MTT assay and lactate dehydrogenase release assay were used to detect uptake of exogenous antigen in immature DCs, the antigen presenting ability in mature DCs and specific cytotoxicity of CTL to leukemic cells, respectively. The DCs with high expression of CD1a, CD86, CD80, HLA-DR, CD54 and CD4 were obtained from marrow and blood of patients with CML. The uptake of FITC-DX was observed in early DCs. There was a potent stimulation to allo-MLR in DCs cultured for 7 - 10 days, and a lightly lower stimulation to auto-MLR. CML-DCs can induce the generation of specific cytotoxic T cells. These results suggest that CML-DCs are functional DCs with the ability to induce anti-leukemia effect.

beta1 Integrin Dysfunction in Adult Chronic Myeloid Leukemia Bone Marrow Cells / 中国实验血液学杂志

According to our previous experiments, Ph(+) chronic myeloid leukemia (CML) cell line K562 cells have defects in beta 1 integrin activation. In order to search the same regularity in Ph(+) CML bone marrow cells, bone marrow mononuclear cells (BMMNC) from 12 cases of Ph(+) CML and 10 cases of normal individuals were studied. Their expression rate of 9EG7 epitope on beta1 integrin post treatment by 8A2 or GM- or G-CSF and cell adhesion ability with soluble fibronectin (FN) were evaluated by flow cytometry; in addition, the effects of CGP57148B, a highly specific ABL tyrosine kinase inhibitor, were observed. Our results showed that 9EG7 expression rate and FN binding rate were very low in all the inactivated cells. The parameter increased markedly post 8A2 activation in both NBMMNCs and CMLBMMNCs, but the degree of increase in CMLBMMNCs was significantly lower than that in NBMMNCs; GM-CSF or G-CSF could significantly increase the parameters in NBMMNCs while had no effects on that in CMLBMMNCs. CGP57148B could increase the beta1 integrin activation potential of CMLBMMNCs but had no effects on that of NBMMNCs. The results indicate that decreased activation potential of beta1 integrin in CMLBMMNCs is the major cause of adhesion defects of Ph(+) CML cells; beta1 integrin functional insufficiency in CMLBMMNCs could not be directly reversed by ABL tyrosine kinase inhibitor CGP57148B.