Clinical Characteristics and Nomogram Model of Nosocomial Infection in Patients with Newly Diagnosed Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To explore the clinical characteristics of nosocomial infection in newly diagnosed multiple myeloma(NDMM) patients, and establish a predictive nomogram model.@*METHODS@#The clinical data of 164 patients with MM who were treated in Shanxi Bethune Hospital from January 2017 to December 2021 were retrospectively analyzed. The clinical characteristics of infection were analyzed. Infections were grouped as microbiologically defined infections and clinically defined infections. Univariate and multivariate regression models were used to analyze the risk factors of infection. A nomogram was established.@*RESULTS@#164 patients with NDMM were included in this study, and 122 patients (74.4%) were infected. The incidence of clinically defined infection was the highest (89 cases, 73.0%), followed by microbial infection (33 cases, 27.0%). Among 122 cases of infection, 89 cases (73.0%) had CTCAE grade 3 or above. The most common site of infection was lower respiratory in 52 cases (39.4%), upper respiratory tract in 45 cases (34.1%), and urinary system in 13 cases (9.8%). Bacteria(73.1%) were the main pathogens of infection. Univariate analysis showed that ECOG ≥2, ISS stage Ⅲ, C-reactive protein ≥10 mg/L, serum Creatinine ≥177 μmol/L had higher correlation with nosocomial infection in patients with NDMM. Multivariate regression analysis showed that C-reactive protein ≥10 mg/L (P＜0.001), ECOG ≥2 (P=0.011) and ISS stage Ⅲ （P=0.024） were independent risk factors for infection in patients with NDMM. The nomogram model established based on this has good accuracy and discrimination. The C-index of the nomogram was 0.779（95%CI: 0.682-0.875）. Median follow-up time was 17.5 months, the median OS of the two groups was not reached (P=0.285).@*CONCLUSION@#Patients with NDMM are prone to bacterial infection during hospitalization. C-reactive protein ≥10 mg/L, ECOG ≥2 and ISS stage Ⅲ are the risk factors of nosocomial infection in NDMM patients. The nomogram prediction model established based on this has great prediction value.

The Prognostic Value of Early Relapse after Autologous Hematopoietic Stem Cell Transplantation in Newly Diagnosed Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To assess the impact of early relapse (ER) after autologous hematopoietic stem cell transplan-tation (AHSCT) on overall survival (OS) for multiple myeloma (MM) patients.@*METHODS@#Clinical data of 37 patients with MM undergoing AHSCT in department of hematology of Shanxi Bethune Hospital from January 2012 to December 2017 were retrospectively analyzed. The effect of ER on OS of patients was analyzed. The effects of international staging system (ISS) staging, cytogenetics, pre-transplant efficacy, minimal residual disease, and age on OS of the patients were also analyzed respectively.@*RESULTS@#Among the 37 patients, 13 cases (35.1%) had ER, and 24 cases (64.9%) had non-ER. 3 patients with ER had extramedullary disease, but none with non-ER showed extramedullary disease. More than or equal to very good partial rate (VGPR) in patients with ER and without ER were 3 cases (23.1%) and 15 cases (62.5%), respectively, and the curative effect of the former was significantly lower than that of the latter (P<0.05). The median follow-up time was 31 (12-96) months, and median OS time was 93 months in all the patients. The median survival time of patients with ER was 17 months, and the median progression free survival was 7 months, both were significantly shorter than 93 months and 38 months of patients with non-ER (P<0.05). Univariate analysis showed that the OS was affected by ER, cytogenetic abnormalities (FISH), and ≥VGPR before transplantation. Multivariate analysis showed that ER was an independent prognostic factor.@*CONCLUSION@#The prognosis of patients with ER after AHSCT in newly diagnosed MM is poor. ER is an independent prognostic factor of survival.

Effect of Serum Free Light Chain Ratio and Normalization Ratio after Treatment on Diagnosis and Prognosis of Patients with Newly Diagnosed Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To evaluate the value of serum free light chain (sFLC) κ/λ ratio (sFLCR) on the diagnosis and prognosis of patients with newly diagnosed multiple myeloma(MM), and explore the effect of sFLCR normalization on the prognosis of patients after 4 courses of induction therapy.@*METHODS@#The clinical data of 43 newly diagnosed MM patients from January 2014 to January 2019 were analyzed retrospectively. Immunoturbidimetry was used to detect the expression levels of sFLC κ and λ. According to the ratio of involved and uninvolved sFLC, using 100 as a boundary, the MM patients were divided into the high ratio group (sFLCR≥100 or ≤0.01) and the low ratio group (0.010.05).@*CONCLUSION@#Patients in the high ratio group at the initial diagnosis have worse renal function, later stage of disease, lower deep remission rate, earlier disease progression, shorter survival time, and worse clinical prognosis.

Effect of Low-Dose Triptolide and Sorafenib Alone and Their Combination on AML Cell Line MV411 and the Pathway of STAT5 / 中国实验血液学杂志

OBJECTIVE@#To investigate the effects of inhibiting proliferation and inducing apoptosis of low-dose triptolide and sorafenib alone or in combination on FLT3-ITD acute myeloid leukemia cell line MV4-11 and STAT5 pathway.@*METHODS@#The MV4-11 cells were treated with low dose triptolide(IC) and sorafenib(IC) alone or in combination for 48 hours. The cell proliferation and inhibition were detected by using CCK-8 kit, the cell apoptosis was detected by flow cytometry, the expression of FLT3,STAT5 in mRNA and protein levels was detected by RT-PCR and Western blot respectively.@*RESULTS@#The treatment of MV4-11 cells with low dose triptolide and sorafenib alone and in combination for 48 hours could inhibit cell proliferation and induce cell apoptosis, moreover the inhibitory rate and apoptotic rate of MV4-11 cells in drug-combination group both were higher than those in single drug group. The mRNA expression and protein expression of FLT3,STAT5 signaling pathway in drug combination group were significantly lower than those in single drug group.@*CONCLUSION@#Low-dose triptolide combined with sorafenib can synergistically inhibit the proliferation and induce the apoptosis of MV4-11 cells, which may be related with the inhibition of FLT3 and STAT5 pathway.

Effect of BRD4 Inhibitor JQ1 on Proliferation Inhibition and Apotosis Induction in Jurkat Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect and possible mechanism of bromo-domain inhibitors (JQ1) on proliferation inhibition and inducing apoptosis of acute T lymphocyte leukemia cell line (Jurkat) .</p><p><b>METHODS</b>Jurkat cell line was treated by JQ1 at different concentrations. MTT was used to detect the cell proliferation inhibition rate. The flow cytometry with AnnexinV-FITC/PI fluorescence staining was used to detect the changes of apoptosis rate, and real-time fluorescent quantitative PCR was used to detect c-Myc/Notch1 gene expression levels.</p><p><b>RESULTS</b>With the increasing of drug concentration and prolonging of time, the inhibitory rate of Jurkat cell growth was enhanced in time-dose dependent manner; Jurkat cells was treated by 0.8, 1.6, and 4 µ mol/LJQ1 for 48 h and 72 h, the cell apoptosis rate was enhanced with the increase of drug concentration and prolonging of time, and the difference was statistically different in comparison with the control group(P<0.05); PCR detection indicated that Notch1 and c-Myc mRNA expression was reduced in 48 h after JQ1 treatment, which was statistically different from the control group,(P<0.05) .</p><p><b>CONCLUSION</b>JQ1 can effectively inhibit the growth of Jurkat cell line, and potentially induce apoptosis through Notch1 and c-Myc gene. Hence JQ1 may be one of new methods used to treat T-ALL.</p>

Mechanism of Notch1 Pathway in SUP-B15 Cell Apoptosis Induced by JQ1 / 中国实验血液学杂志

<p><b>OBJECTIVE</b>The study was aimed to investigate the possible mechanism of Notch1 pathway in apoptosis of Ph(+) human ALL Cells(SUP-B15 cells) induced by bromodomain inhibitors JQ1.</p><p><b>METHODS</b>The SUP-B15 cells were treated with different concentrations of JQ1 for different times. The cell proliferation was analyzed with cytotoxicity test(MTT method). Cell cycle was detected by fluorescence microscopy and flow cytometry. The mRNA expression of MIS2, Notch1, Hes1, BCR-ABL in Notch1 pathway was detected by real-time quantitative PCR.</p><p><b>RESULTS</b>JQ1 0-4 µmol/L could significantly inhibit the viability of SUP-B15 cells treated in does-and time-dependent manner. After SUP-B15 cells were treated with 1,2,4 µmol/L JQ1 for 48 h, the JQ1 could induce S cycle arrest in does-dependent manner which was statistical different from the control at the same time (P<0.05). MIS2, Notch1, Hes1, BCR-ABL mRNA expression was down-regulated by JQ1 which was statistical different from the control (P<0.05).</p><p><b>CONCLUSION</b>The JQ1 can effectively inhibit the growth and proliferation of SUP-B15 cells and the Notch1 pathway may be one of the important apoptosis mechanisms in Ph(+) ALL cells induced by JQ1.</p>

Apoptosis-inducing effect of tetrandrine and imatinib on K562/G01 cells and its related mechanism / 中国实验血液学杂志

This study was purposed to explore the apoptosis-inducing effect of tetrandrine (Tet) and imatinib (IM) alone or both combined on K562/G01 cells and their mechanism. MTT assay was used to detect the inhibitory effect of drugs on cell growth, flow cytometry was used to detect the cell cycle and apoptosis rate. The expression of caspase-3/BCL-2 mRNA was determined by real time-PCR, and the expression of caspase-3/BCL-2 protein was assayed by Western blot. The results showed that after being treated by 1.0 µmol/L IM or 1.5 µmol/L Tet alone and combination of these two drugs for 48 h, the inhibitory rate was (22.74 ± 0.05)%, (20.34 ± 0.57)% and (44.28 ± 0.60)%, respectively, suggesting that inhibitory effect of two drug combination was more obvious. The arrest of cell cycle at G1/S phase could be observed after Tet treatment. Early apoptosis rate was (7.81 ± 0.16) %, (14.10 ± 0.28) % respectively after being treated by combination of 1.5 µmol/L and 3.0 µmol/L Tet with 1.0 µmol/L IM. After being treated with Tet alone, FQ-PCR and Western blot showed that the expressions of caspase-3 mRNA and caspase-3 protein were up-regulated, the expressions of BCL-2 mRNA and protein were down-regulated, the effect of both drug combination was more significant. It is concluded that IM or Tet alone can induce apoptosis of K562/G01. Combination of IM with Tet shows obvious synergistic effect, mechanism of which may associate with up-regulation of caspase-3 mRNA and protein expressions, and down-regulation of BCL-2 mRNA and protein expressions.

Effect of bortezomib on reverse multidrug resistance and XIAP expression in imatinib-resistant primary cells of chronic myeloid leukemia in blastic crisis / 中国实验血液学杂志

This study was aimed to explore the effects of proteasome inhibitor bortezomib on the drug sensitivity of imatinib-resistant primary cells in blastic phase of chronic myeloid leukemia (CML) and the expression of XIAP. MTT method was used to detect the inhibitory effect on cell growth, flow cytometry was used to assay the apoptosis and P170 expression, and RT-PCR was used to monitor the expression of XIAP mRNA. The results showed that the effect of imatinib or bortezomib alone showed an inhibitory effect on MNC in time-and dose-dependent manner; 5 and 10 nmoL/L bortezomib combined with imatinib could significantly enhance the sensitivity of mononuclear cells to imatinib. The increase of apoptosis rate, and the decrease of P170 expression could be observed by flow cytometry during treatment with bortezomib. The over-expression of XIAP could be down regulated by bortezomib. It is concluded that the bortezomib could inhibit primary cells of leukemia and enhance sensitivity of CML primary cells to imatinib.The bortezomib may increase the cell apoptosis by inhibition of XIAP expression, so as to provide the experiment evidence to spread bortezomib for the clinical treatment of chronic myeloid leukemia.

Triptolide inhibits proliferation and induces apoptosis of imatinib resistant K562/G01 cells / 中国实验血液学杂志

This study was aimed to explore the inhibitory effect of triptolide on proliferation and inducing apoptosis effect of K562/G01 cells and their possible mechanism. MTT assay was used to detect the effect of imatinib or triptolide alone and their combination on K562/G01 proliferation; the cell cycle, apoptosis rate, P-gp protein expression were detected by flow cytometry (FCM); the expression of P-gp was assessed by Western blot; the BCR/ABL gene expression was assayed by real time quantitative PCR. The results showed that triptolide could enhance the effect of imatinib on proliferation inhibition and apoptosis of K562/G01, arrested the cell cycle in G1 phase, down-regulated the expression of BCR/ABL gene and P-gp protein. It is concluded that triptolide induces K562/G01 cell proliferation inhibition and apoptosis, the mechanism may be related to cell cycle arrest, decrease of P-gp protein expression, inhibition of BCR/ABL gene expression.

Prophylactic effect of TLR5 agonist flagellin on acute graft versus host disease after allogeneic hematopoietic stem cell transplantation and its mechanism / 中国实验血液学杂志

This study was aimed to investigate the prophylactic effect of Toll like receptor (TLR)5 agonist flagellin on acute graft versus host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and its possible mechanism. The animal model with allo-HSCT aGVHD was established by using purebred mice (male mouse C57BL/6 as donor, female mouse BALB/c as recipient) with complete-unidentical major histocompatibility antigen. The recipient mice were randomly divided into 3 groups: group 1 in which mice were injected with high purity (95%) flagellin before and after allo-HSCT respectively, group 2 in which mice received allo-HSCT without injection of flagellin, group 3 in which mice were radiated alone. The aGVHD features of mice in group 1 and 2 were observed and compared. The results showed that the typical symptoms of aGVHD appeared in transplanted mice. The death peak of mice in group 2 appeared at day 4-5 after transplantation. The aGVHD symptoms were obviously alleviated and the mean survival time was prolonged significantly in mice group 1 as compared with mice in group 2 (P < 0.05). The comparison of WBC count in peripheral blood of mice in 3 groups before transplantation showed no significant difference (P > 0.05), while WBC count of mice in group 1 and 2 showed the significant difference at days 14 and 21 after transplantation (P < 0.05). The pathological appearances of aGVHD in mice of group 1 were obviously reduced as compared with mice in group 2. The flow cytometric detection of Treg cell/CD4(+) T cell levels at different time before and after transplantation demonstrated that the Treg cell level in mice of group 1 at weeks 2-4 after transplantation significantly increased as compared with mice in group 2 (P < 0.05). It is concluded that flagellin can effectively prevent the aGVHD occurrence after allo-HSCT, reduce the symptoms and pathological changes of aGVHD, obviously prolong mean survival time of mice in group 1. The mechanism of flagellin effect may be associated to increase of Treg cell level in mice after allo-HSCT.

Effect of low-dose decitabine in combination with imatinib mesylate in K562 cells / 白血病·淋巴瘤

Objective Imatinib mesylate (IM) is the most active agent in treating chronic myeloid leukemia (CML). 5-Aza-2-deoxycytidine (DAC) is a cytosine analogue that inhibits DNA methylation and the activity in myeloid leukemia. Therefore,we investigated combining these two drugs in human leukemia cell line K562. Methods The effects of IM and DAC was examined in K562 cells including cell viability using MTT method,cell cycle phase and cell death using flow cytometric (FCM),and the expression of bcr-abl mRNA by RT-PCR. Results Both DAC and IM resulted in time and concentration-dependent induction of cell death. DAC and IM in combination produced a greater inhibition of growth against K562 cells (F =43.947,165.580,321.193,296.101,P<0.05). The main effect and interaction between two drugs was statistically significant (F = 202.759,168.457,417.538,P <0.001) after 24 h,48 h,72 h and a greater reduction in expression of bcr-abl mRNA than either agent alone. The difference was statistically significant (F =71.981,P <0.05). The number of G1 phase cells were increased significantly when induced by single agent. 48 h incubation with IM 0.2 μmol/L alone or combined with DAC 4 μmol/L showed 6.7 %,8.4 % pre-apoptosis cells,respectively. After incubation for 48 h with DAC 4 μmol/L,the expression of mRNA were decreased by 14 %,IM 0.2 μmol/L showed 40 % reduction,and combination group were significantly depressed for the mRNA expression by 60 %. Conclusion The combination of DAC and IM showed synergistic effects on cell death in K562 cells. These data suggested that DAC used in combination with IM has clinical potential in the treatment of chronic myeloid leukemia.

Effect of bortezomib on the drug sensitivity of imatinib resistant K562/G01 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of bortezomib (BOR) on the drug sensitivity of imatinib-resistant chronic myeloid leukemia cell line K562/G01 cell and its mechanism.</p><p><b>METHODS</b>MTT assay was used to detect the inhibition effect of cell growth, flow cytometry to cell cycle, and real time-PCR to the expression of COX-2 and mdr1 mRNA.</p><p><b>RESULTS</b>Combination of 10 and 20 nmol/L BOR with imatinib could significantly enhance the sensitivity of K562/G01 to imatinib, the reverse factor was 1.83 and 2.72-fold respectively. Cell cycle arrested at G(2)/M phase could be observed by flow cytometry on BOR treatment. The over-expression of COX-2 and mdr1 could be down-regulated by BOR.</p><p><b>CONCLUSIONS</b>BOR can enhance the imatinib sensitivity of imatinib resistant K562/G01 cell. The mechanism may be related to cell cycle phase arrested at G2/M and down-regulation of COX-2 and mdr1 expression.</p>

Relationship between CD4(+)CD25(+) regulatory T cells, IL-2, TGF-beta and acute graft-versus host disease after allogeneic hematopoietic stem cell transplantation / 中国实验血液学杂志

This study was aimed to investigate the relationship between CD4(+)CD25(+) regulatory T cells, IL-2, TGF-beta and acute graft-versus host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The percentage of peripheral blood CD4(+)CD25(+) Treg cells in CD4(+) T cells of 13 patients with hematological malignancies after allo-HSCT were detected by flow cytometry; serum levels IL-2 and TGF-beta in these patients were measured by ELISA. The results indicated that all the patients achieved engraftment. 5 patients developed aGVHD of grade I-II, 4 patients developed aGVHD of grade III-IV. The percentage of peripheral blood CD4(+)CD25(+) Treg cells out of CD4(+) T cells in patients without aGVHD was higher than that in patients with aGVHD (p < 0.05); the serum level of IL-2 in patients without aGVHD was lower than that of patients with aGVHD (p < 0.05); the serum level of TGF-beta in patients without aGVHD was higher than that of patients with aGVHD (p < 0.05). It is concluded that CD4(+)CD25(+) Treg cell level and the serum level of IL-2 and TGF-beta all are related to incidence and severity of aGVHD. These factors may be used as indicators for early evaluating and monitoring aGVHD after allo-HSCT.

Arsenic trioxide combined with buthionine sulfoximine enhances apoptosis in multidrug-resistant human leukemia K562/ADM cells in vitro / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the apoptosis-inhancing effect of the combination of arsenic trioxide (As2O3 ) and buthionine sulfoximine (BSO) on multidrug-resistant human leukemic K562/ADM cells, to compare the effect of As2O3 alone and As2O3 combined with BSO and As2O3 alone, and to determine the effect of intracellular GSH content on this treatment.</p><p><b>METHODS</b>As2O3 was used in a dose of 0.5 micromol/L, 2.0 micromol/L and 5.0 micromol/L, respectively, and BSO was used in a dose of 100 micromol/L in the culture of multidrug-resistant human leukenic K562/ADM cells. The cell proliferation activity was assessed with MTT assay. The cell apoptosis was detected by flow cytometry using Annexin-V and propidium iodide (PI) staining. Intracellular GSH content was measured using glutathione assay kit by spectrophotometry.</p><p><b>RESULTS</b>After the GSH contents were reduced by the combination of arsenic in clinic dose (0.5, 2 micromol/L) and BSO (100 micromol/L), respectively, the K562/ADM cell proliferation activity was obviously inhibited and the cell apoptosis-inducing effect was advanced in 24 hours. In 48 and 72 hours, the effect of the combination group (clinic dose arsenic group) was significantly stronger than that of clinic dose arsenic alone group and the high dose arsenic alone group.</p><p><b>CONCLUSION</b>The cell apoptosis-inducing effect of arsenic in combination of BSO on multidrug resistant human leukemia K562/ADM cells is significantly enhanced in comparison with that of arsenic alone. The reduction of intracellular glutathione content is closely correlated with this apoptosis-enhancing effect.</p>

The effect of arsenic trioxide (As2O3) combined with BSO on K562/ADM cell and its mechanisms / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the apoptosis-induction, P-glycoprotein (P-gp) and mdr1 mRNA inhibition effects of arsenic trioxide (As2O3) and buthionine sulfoximine (BSO) on multidrug-resistant cell line K562/ADM cells, and to determine the relationship between intracellular GSH content and arsenic effect.</p><p><b>METHODS</b>K562/ADM cells were treated with arsenic (0.5, 2.0, 5.0 micromol/L) alone or combined with BSO (100 micromol/L). The cell proliferating capacity was assessed with MTT assay, and cell apoptosis by Annexin V and propidium iodide (PI) staining. Intracellular GSH contents were measured using a glutathione assay kit by spectrophotometry. P-gp expression was determined by flow cytometry, and mdr1 mRNA expression by semi-quantitative RT-PCR.</p><p><b>RESULTS</b>The GSH contents in K562/ADM cell was (81.13 +/- 3.91) mg/g protein. After the GSH contents were degraded by BSO, the K562/ADM cell proliferating capacity was obviously inhibited and the cells were induced apoptosis in 24 hours by the combination of clinic dose arsenic group (0.5, 2.0 micromol/L) and BSO (100 micromol/L). The cell apoptosis rates at 48 hours in arsenic alone group and combination group were (59.29 +/- 6.01)% and (65.06 +/- 8.29)%, and at 72 hours were (82.15 +/- 9.28)% and (92.72 +/- 9.41)% retrospectively. At 48 hours, the mdr1 mRNA inhibition effect of the combination group was obviously stronger than that of high dose arsenic alone group. At 72 hours, the P-gp inhibition effect of the combination group (clinic dose arsenic group, 0.5, 2.0 micromol/L) was obviously stronger than that of high dose arsenic alone group (5.0 micromol/L).</p><p><b>CONCLUSION</b>The intracellular GSH contents are closely correlated with the arsenic effect. The combination of conventional dose arsenic and BSO significantly induces K562/ADM cell apoptosis and inhibits P-gp and mdr1 mRNA expression in the cells.</p>

Clinical studies on expressions of Fas and mdr-1 in acute leukemia and their correlations / 中国实验血液学杂志

To explore the Fas and mdr-1 expression and their correlation in multidrug resistance (MDR), Fas and mdr-1 expressions of bone marrow from 59 patients with newly diagnosed AL before therapy and after complete remission were detected by direct immunofluorescence with flow cytometry and semi-quantitative RT-PCR, respectively. The results showed that in newly diagnosed AL patients, Fas expression in AML was higher than in ALL (P < 0.05), mdr-1 expression in AML and ALL had no difference (P > 0.05), the expressions of Fas and mdr-1 had correlation (r = -0.282, P < 0.05) in AL; the results of simple-variable and multivariable COX survival factor model analysis suggested that Fas and mdr-1 expressions were prognostic factors for the effect of therapy and survival. Log rank test, comparing the groups of Fas(+) with Fas(-), mdr-1(+) with mdr-1(-), demonstrated that the CR rates and median remission time of every two groups had significant difference. It is concluded that in AL, Fas and mdr-1 expressions have high correlation with the effect of treatment, Fas expression probably is one of the favorable prognostic factors, mdr-1 is an unfavorable prognostic and less effective factor.

Research progress on mechanisms of arsenic trioxide in treating lymphoma / 肿瘤研究与临床

Now the treatment of lymphoma mainly uses combined chemotherapy and radiotherapy.Es- pecially hemopoietic stem cell transplantation is effective in the treatment to the patients with refractory and later stage.During the recent years the domestic and foreign scholars have obtained the encouraging results with arsenic trioxide to treat lymphoma.This article has reviewed the experimental study and the clinical re- search progress about arsenic trioxide in application to lymphoma cells.