Efficacy of CLAE Chemotherapy Regimen Followed by Allogeneic Hematopoietic Stem Cell Transplantation in Patients with Relapsed/Refractory Acute Leukemia / 中国实验血液学杂志

OBJECTIVE@#To observe the efficacy and safety of CLAE intensive chemotherapy followed by allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with relapsed/refractory acute leukemia (R/R AL).@*METHODS@#CLAE regimen [cladribine 5 mg/(m2·d), d 1-5; cytarabine 1.5 g/(m2·d), d 1-5; etoposide 100 mg/(m2·d), d 3-5] followed by allo-HSCT was used to treat 3 R/R AL patients. The patients received CLAE chemotherapy in relapsed or refractory status and underwent bone marrow puncture to judge myelodysplastic state. After an interval of 3 to 5 days, followed by preconditioning regimen for allo-HSCT [fludarabine 30 mg/(m2·d), d -7 to d -3; busulfan 0.8 mg/kg q6h, d -6 to d -3 or d -5 to d -2. If the bone marrow hyperplasia was not active and the blasts were less than 10%, busulfan should be used for 3 days. If the bone marrow hyperplasia was active and the blasts were more than 10%, busulfan should be used for 4 days]. Cyclosporin A, mycophenolate mofetil and short-term methotrexate were used for graft-versus-host disease (GVHD) prevention. After transplantation, the status of minimal residual disease (MRD) and bone marrow chimerism were regularly monitored in all 3 patients, and demethylation drugs or dasatinib were used to prevent recurrence 3 months after transplantation.@*RESULTS@#2 patients with t(11;19) translocation and relapse/refractory acute myeloid leukemia recurred within 6 months after induction of remission, and received intensive chemotherapy with CLAE regimen followed by haploidentical allo-HSCT and unrelated donor allo-HSCT, respectively. The two patients both relapsed 6 months after transplantation, then achieved complete remission by donor lymphocyte infusion, interferon, interleukin-2 and other methods, and disease-free survival was 2 years after transplantation. The other patient was chronic myelogenous leukemia who developed acute lymphoblastic leukemia during oral administration of tyrosine kinase inhibitor, accompanied by T315I and E255K mutations in ABL1 kinase region and additional chromosomal abnormalities. After morphological remission by induction chemotherapy, central nervous system leukemia was complicated. Intensive chemotherapy with CLAE regimen followed by sibling allo-HSCT was performed in the positive state of MRD. The patient relapsed 3 months after transplantation, and achieved remission after chimeric antigen receptor T-cell (CAR-T) therapy, however, he died 5 months after transplantation because of severe cytokine release syndrome (CRS) and GVHD.@*CONCLUSION@#CLAE regimen followed by allo-HSCT may be an effective salvage treatment option for R/R AL patients to prolong the overall survival.

Effect of CXCR4 on the Treatment Response and Prognosis of Carfilzomib in Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To explore the effect of CXCR4 on the treatment response and prognosis of Carfilzomib (CFZ) in multiple myeloma.@*METHODS@#Dataset GSE69078 based on microarray data from two CFZ-resistant MM cell lines and their corresponding parental cell lines (KMS11-KMS11/CFZ and KMS34-KMS34/CFZ) were downloaded from Gene Expression Omnibus (GEO). Differentially expressed genes (DEGs) were identified, and Protein-protein interaction (PPI) network was established to identify the key genes involved in CFZ resistance acquisition. Finally, the prognostic roles of the CFZ risistance key genes in MM using MMRF-CoMMpass data study was verified.@*RESULTS@#44 up-regulated and 46 down-regulated DEGs were identified. Top 10 hub genes (CCND1, CXCR4, HGF, PECAM1, ID1, HEY1, TCF4, HIST1H4J, HIST1H2BD and HIST1H2BH) were identified via Protein-protein interaction (PPI) network analysis. The CoMMpass data showed that high CXCR4 expression showed correlation to relative higher relapse and progress rates and the overall survival was significant decreased in high CXCR4 patients (P=0.013).@*CONCLUSION@#CXCR4 perhaps plays a crucial role in CFZ acquired resistance, which might help identifying potential CFZ-sensitive patients before treatment and providing a new therapeutic target in CFZ-resistant MM.

The renal protective effect of saxagliptin combined with metformin on type 2 diabetic mice and its relationship with redox balance / 中国药理学通报

Aim To evaluate the reno-proteetion of saxagliptin combined with metformin in mice with T2DM anrl its relationship with redox balance.Meth¬ods C57BL/6J mice were fed with high fat diet and injected with low dose STZ intraperitoneally to establish T2DM mouse model.Then they were randomly divided into T2DM group, glibenclamide group ( Gli group), metformin group ( Met group) , saxagliptin group ( Sax group) and saxagliptin + metformin group ( S + M group) , and normal control group ( NC group) with 8 mice in each group.Eight weeks after intervention the mice were weighed.Blood, urine and renal tissue sam¬ples were collected to measure GHbA,c, FBG, Alb, 8-OHdG, 8-iso-PG, SOD, GSH, GSSG and Ucr.The pathological morphology of renal tissues in each group was observed.Results Saxagliptin combined with metformin reduced significantly the levels of Alb/Ucr ( UACR) , 8-0HdG/Ucr( UOCR) , 8-iso-PG/Ucr( UP- CR) , increased the activity of SOD and GSH/GSSG ratio, and improved the pathological changes of renal tissues, which were superior to those in Met group and Sax group.Conclusions Saxagliptin combined with metformin have a synergistic protective effect on the kidneys of type 2 diabetic mice.The mechanism is partly related to alleviating oxidative stress and impro¬ving redox balance in vivo.

Efficacy of Micro-Transplantation Consolidation Therapy for Patients with Acute Myeloid Leukemia after Complete Remission / 中国实验血液学杂志

OBJECTIVE@#To investigate the efficacy and safety of micro-transplantation in acute myeloid leukemia (AML).@*METHODS@#The clinical data of 13 adult AML patients who received micro-transplantation as consolidation therapy from July 2014 to October 2019 was retrospectively analyzed, and the adverse reactions and efficacy of micro-transplantation were followed up.@*RESULTS@#Eight patients received micro-transpantation were still in complete remission, 5 patients relapsed after micro-transplantation, 1 of them received umbilical cord blood micro-transplantation after remission by reinduction, and all of the 13 patients have survived till now. The median overall survival time was 13 months, and the median relapse-free survival time was 12 months. All 13 patients developed grade 2-4 hematological adverse reactions. The median recovery time of neutrophils and platesets was 13 (11-15) and 15 (13-17) days, respectively. None of the 13 patients developed acute or chronic graft versus host disease. Twelve patients suffered from different infections, however, there were no serious organ function injury complications happened.@*CONCLUSION@#The micro-transplomtation of HLA-incompatible stem cells derived from peripheral blood or umbilical and blood is an effective regimen for the consolidation therapy of AML, especially for the patients suffered from low and moderate risk of AML or the aged AML patients.

Research Advance on the Role of Spleen Tyrosine Kinase Inhibitors in Hematologic Malignancies / 中国实验血液学杂志

Abstract　　Spleen tyrosine kinase (SYK) is not only a key kinase in the B-cell receptor (BCR) signaling pathway, but also a critical component of other signal transduction pathways such as Fc receptor, complement receptor and integrin. Abnormal activation of SYK closely related to the occurrence and development of hematological malignancies, thus targeting SYK has become a research hotspot. Several SYK inhibitors including Fostamatinib, Entospletinib and Cerdulatinib were being evaluated in clincal trials. As a second generation SYK inhibitor, Entospletinib has achieved good efficacy in lymphoid and myeloid hematologic tumors. Furthermore, Entospletinib can significantly relieve hematopoietic stem cell transplantation(HCT) related graft versus host disease (GVHD). In this review the role of SYK inhibitors in treatment of hematological malignancies is summarized brifely.

Effects of oxygen-enriched negative pressure wound therapy in improving wound microenvironment to tissue proliferation and vascularization / 医学研究生学报

ObjectiveIt is indefinited that oxygen-enriched negative pressure wound therapy, namely negative pressure wound therapy combined with topical oxygen therapy (NPWT+TOT), improve the effects of wound microenvironment in tissue proliferation and vascularization. The objective is to discuss effects of oxygen-enriched negative pressure wound therapy in improving wound microenvironment to tissue proliferation and vascularization.MethodsTo select sixty-four patients in the outpatient wound care center of the eastern theater general hospital from January to October, 2019, which were randomly divided into the experimental group (NPWT+TOT) and the control group (NPWT), 32 cases in each group. The patients were treated with oxygen-enriched negative pressure wound and negative pressure wound respectively for 2 weeks to observe the changes of wound temperature and PH before and during intervention. Bacterial culture and immunohistochemical staining which were made from wound secreta and wound bed tissues to observe bacterial culture results, tissue proliferation activity and microvascular density before intervention and 14 days after intervention. After the intervention, the patients were treated by standard wet therapy and followed up to wound healing or 3 months after the intervention to observe the wound healing rate and wound healing time.ResultsAfter two weeks' continuous intervention, wound temperature of patients increased and PH value decreased significantly between the experimental group and the control group. Meanwhile, the intervention group was more effective, and there were significant differences between the two groups (P<0.05). The positive rate of bacterial culture after intervention in the experimental group and the control group was 26.67% and 41.38% respectively, with no statistically significant difference (P=0.233). Compared with the control group, the increase of tissue activity and microvascular density in the experimental group was more significant (P<0.05). After three months' follow-up, the wound healing rate of the experimental group was increased by 12.5% compared with the control group, and the average wound healing time was shortened by 9.2 days.ConclusionOxygen-enriched negative pressure wound therapy can improve wound microenvironment, reduce the positive rate of wound bacterial culture, improve the proliferation activity of wound tissue and degree of vascularization, and promote wound healing.

Reaserch Advance on Bruton Tyrosine Kinase Inhibitors in the Treatment of B-Cell Tumors--Review / 中国实验血液学杂志

Abstract　　In recent years, development of the targeted drugs according to the biological characteristics of tumors have provided more treatment options for tumor patients. It was found that the overactivation or abnormality of B cell receptor (BCR) signal pathway closely related to the occurrence and development of various B cell tumors, such as chronic lymphocytic leukemia (CLL) and mantle cell lymphoma (MCL). As a key kinase in the BCR pathway, BTK inhibitors have obvious anti-tumor effect when its activity is being inhibitered. Currently, BTK inhibitors developed include the first-generation Ibrutinib and the second-generation Acalabrutinib, which can be targeted at the inhibition of BTK and its downstream signaling pathway, and have important therapeutic value for a variety of B-cell tumors, such as CLL and partial non-Hodgkin's lymphoma (NHL). However, its side effects and drug-resistance also gradually emerged, effective drug combination therapy has shown a certain clinical activity. This reviews summarizes briefly the mechanism and status of BTK inhibitors in the treatment of various B-cell tumors.

Effect of Puerarin in Reducing Insulin Resistance in HepG2 Cells via PI3K/Akt/GSK-3β Signal Transduction Pathway / 中国实验方剂学杂志

Objective: To observe the effect of puerarin on phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and glycogen synthase kinase-3β (GSK-3β) in insulin resistant HepG2 cells. Method: HepG2 cells were treated with palmitic acid 0.5 mmol·L-1 and insulin 9×10-4 U·L-1 to induce insulin resistant condition for 24 h. Cell viability was detected by methyl thiazolyl tetrazolium (MTT) assay to determine the concentration of puerarin. This experiment included normal control group, model control group and puerarin groups of different doses (40, 80, 160,320 μg·L-1). Glucose detection kit was used to detect the content of glucose in cell culture supernatant. Tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) levels in supernatant of cell culture medium were detected by enzyme-linked immunosorbent assay (ELISA). Hepatic glycogen assay kit was used for detecting the hepatic glycogen content in HepG2 cells. Western blot was applied to detect protein expression levels of PI3K, Akt, p-Akt, GSK-3β and p-GSK-3β. Result: Compared with those in the normal control group, the glucose consumption rate was significantly down-regulated in HepG2 cells in the model control group (PPα and IL-6 were increased in supernatant of cell culture medium (PPβ protein expression was up-regulated (PPα and IL-6 were reduced in supernatant of cell culture medium (PPβ protein expression was down-regulated, but its phosphorylation inactivation was increased (PConclusion: Puerarin alleviates the insulin resistance of HepG2 cells by strengthening the PI3K/Akt/GSK-3β signal transduction process and increasing the glycogen content in hepatocytes.

Comparison of the effectabout improved negative pressure wound therapy with filling dressing for treating traumatic wounds / 医学研究生学报

Objective The purpose of this study is to use nano-silver dressing as filling dressing for negative pressure wound therapy, and to observe the effect of treating traumatic infected wounds, so as to provide a basis for optimizing negative pressure wound therapy technology. Methods Eighty patients with physical traumatic wounds were enrolled in the outpatient wound care center. They were randomly divided into the intervention group and the control group (n=40 in each group). In the intervention group, the filling dressing for negative pressure wound therapy used nano silver dressing. In the control group, the standard negative pressure wound therapy with normal saline gauze as filling dressing was adopted. All patients were treated with negative pressure for at least 14 days, and then were treated with moist wound therapy until followed up for wound healed. The wound volume reduction rate was the main outcome indicator 14 days after intervention in the two groups. The bacterial positive rate and the wound healing rate at the end of 3 months follow-up, and final healing time were the secondary outcome indicators. Results 36 cases in the control group and 40 cases in the intervention group completed the expected negative pressure wound therapy time and follow-up. The baseline data of the two groups had no difference. The wound volume reduction rate (70.95±20.73)% in the intervention group after 14 days of treatment was significantly higher than that in the control group (64.42±22.33)% (P< 0.05), and the bacterial positive rate (20%) was lower than that in the control group (44.44%)(P< 0.05). At the end of the follow-up, the healing rate of the intervention group (97.50%) was higher than that of the control group (66.67%)(P<0.001). The healing time of the intervention group（50.85±15.81d）was shorter than of the intervention group (62.58±16.18d)(P<0.05). Conclusion Improving the filling dressing for negative pressure wound therapy can effectively reduce the volume of traumatic wound and the positive rate of pathogenic bacteria, and help to control wound infection and improve wound healing outcomes.

Therapeutic Response and Prognosis of Adult Acute Myeloid Leukemia with Chromosome Karyotype Abnormalities / 中国实验血液学杂志

OBJECTIVE@#To investigate the efficacy and prognosis of acute myeloid leukemia (AML) patients with chromosome karyotype abnormalities.@*METHODS@#The clinical features and treatment responses of 91 patients with AML were collected and analyzed retrospectively. The efficacy and survival rate of the AML patients with normal and abnormal chromosome karyotype were compared.@*RESULTS@#Chromosome translocations and monosomal karyotypes were the main heterogeneity of AML. There was no significant difference in complete remission rate and overall response rate between the normal and abnormal karyotype groups, but the recurrence rate was higher in abnormal karyotype group. There was no significant difference in response of AML patients received the standard "3+7 regimen" and pre-excitation chemotherapy in the treatment of normal and abnormal karyotype groups. The relapse free survival time (RFS) was longer in the normal karyotype group, but there was no significant difference in overall survival time (OS).@*CONCLUSION@#The abnormal karyotype of AML is an independent prognostic factor, monosomal karyotype shows a poor prognosis, and the recurrence rate in AML patients with monosomal karyotype is higher.

Relationship Between Helicobacter Pylori Infection and Carotid Intima Thickness by Meta–analysis / 中国循环杂志

Objectives: To investigate the relationship between helicobacter pylori (Hp) infection and carotid intima thickness. Methods: We searched the databases of Pubmed, Embase, CNKI, VIP and Wanfang for published studies on the relationship between Hp infection and carotid intima thickness in China and abroad. 2 reviewers independently identified and enrolled the articles, extracted specific data and assessed the quality of reference by Jadad scale. Meta-analysis was conducted by Stata 12.0 Software. Results: A total of 9 case-control studies were enrolled, all the reference quality scores≥4 which included 694 subjects with Hp positive and 676 with Hp negative. Meta-analysis indicated that carotid intima thickness was higher in Hp positive subjects than Hp negative subjects, standard mean difference (SMD)=0.803, 95% CI(0.686-0.919), P<0.05. Age related study presented that carotid intima thickness was higher in Hp positive subjects than Hp negative subjects at the age≥60 years'population, SMD=1.296, 95%CI (1.125-1.467), P<0.05; carotid intima thickness was also higher in Hp positive subjects than Hp negative subjects at the age<60 years'population, SMD=0.379, 95%CI (0.220-0.537), P<0.05. Conclusions: Hp infection has been related to the thickness of carotid intima and it could make carotid artery intima thickening.

Effects of metformin on SIRT1 expression of renal tissues in type 2 diabetic rats / 中国药理学通报

Aim To observe the effects of metformin (MET) on the silencing regulatory protein 1 (SIRT1) mRNA and protein expression in renal tissues of type 2 diabetic mellitus (T2DM) rats, and explore its reno-protective mechanisms. Methods Thirty model T2DM rats were randomly divided to glibenclamide in-tervention group (GLY group, 5 mg·kg-1·d-1), metformin intervention group (MET group,300 mg· kg-1· d-1) and diabetic control group (T2DM group),and 10 rats with normal glucose tolerance were used as normal control(NC group). After 8 weeks, HbA1c,blood glucose (BG), urea nitrogen (BUN), urinary albumin, creatinine and glomerular basement membrane thickness (GBMT) were detected. The ex-pression of SIRT1 protein in renal tissues was detected by immunohistochemistry, the expression of SIRT1 mRNA in renal tissues was detected by real-time PCR, and urinary SIRT1 protein was detected by ELISA. Results At the end of 8 weeks, the levels of BG,HbA1c,urinary albumin/urinary creatinine(UACR), urinary SIRT1/urinary creatinine (USIR) and GBMT in MET and GLY groups were significantly lower than those in T2DM group (P<0.05). There was no sig-nificant difference in BG, HbA1c and GBMT between MET group and GLY group (P>0.05). The expres-sions of SIRT1 mRNA and protein in MET group were significantly lower than those in NC group (P <0.05), but higher than those in T2DM group (P <0.05). The UACR, expression of SIRT1of renal tis-sues in MET group was higher than that in GLY group (P<0.05),but urinary SIRT1 protein was lower than that in GLY group (P <0.05). Conclusion Met-formin can increase the expressions of SIRT1 in renal tissues of T2DM rats,which may be related to its renal protection.

Establishment of Drug-resistant Acute Lymphoblastic Leukemic Cell Lines and Their Resistance Mechanism / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To establish the adriamycin(ADR)-resistant ALL cell lines and to investigate their drug-resistan mechanisms.</p><p><b>METHODS</b>The drug-resistant cell lines SUP-B15/ADR and RS4;11/ADR were derived by exposing the parental cells [SUP-B15(Ph) and RS4;11(Ph)] to the ascending concentrations of ADR. The cell viability was detected by CCK-8 method. The expression of P-gp was examined by Western blot, and RT-qPCR was performed to detect the expression of MDR1.</p><p><b>RESULTS</b>The drug-resistant cell lines SUP-B15/ADR and RS4;11/ADR were successfully established, their resistance indexes were 14.088±0.763 and 10.473±1.024, respectively. After the cryopreserved SUP-B15/ADR and RS4;11/ADR cells were resuscitated, their survival rates were 88.4±1.2% and 89.3±1.6% respectively, while their resistance indexes were 13.976±0.967 and 10.342±0.846 respectively (P>0.05). When the drug-resistant cells were cultured in the medium without ADR for 1 month, their drug-resistance indexes dropped down to 12.893±1.255 and 9.327±0.321 respectively(P<0.05). Drug-resistant cell lines had the cross-resistance to cytarabine and etoposide. The expression of P-gp and MDR1 in drug-resistant cells was significantly higher than that in wild-type cells.</p><p><b>CONCLUSION</b>Two drug-resistant ALL cell lines have been successfully established by exposing to the ascending concentration of ADR. The over-expression of MDR1 and P-gp in drug-resistant cells may be one of the mechanisms underlying the drug resistance.</p>

Effects of PCI-32765 and Dasatinib on the Acute Lymphoblastic Leukemic Cells and Their Mechanisms / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of Btk inhibitor (PCI-32765) and BCR-ABL tyrosine kinase inhibitor (Dasatinib) on proliferation and apoptosis of acute lymphoblastic leukemia (ALL) cell lines (Sup-B15, RS4;11) and the possible mechanism.</p><p><b>METHODS</b>RS4;11 and Sup-B15 cells were treated with PCI-32765 and Dasatinib, the cell proliferation and apoptosis were detected by CCK-8, the Btk and other apoptotic proteins were detected by Western blot.</p><p><b>RESULTS</b>PCI-32765 could inhibit the proliferation of RS4;11 and Sup-B15 cells in a dose-dependent manner, Sup-B15 cells were more sensitive to PCI-32765 than RS4;11 cells, their ICwere 3 µmol/L and 8 µmol/L respectively, the difference between them was statistically significant (P<0.05). Dasatinib also could inhibit the proliferation of RS4;11 cells and Sup-B15 cells in a dose-dependent manner. The ICwas 5 µmol/L and 5 nmol/L, respectively, the difference between them was statistically very significant (P<0.01), and the inhibitory effect was enhanced by the combination of Damatinib with the PCI-32765(P<0.05). The cell survival rate decreased gradually in PCI-32765 or Dasatinib alone group and the combination group at the different time-point (8, 12, 24, 36, 48 and 72 h), the 2 drugs showed a synergistic effect on cells in a time-dependent manner. After being treated with PCI-32765 and Dasatinib, the RS4;11 and Sup-B15 cells showed that cell shrinkage, increase of cytoplasmic density, nuclear pyknosis, deviation and karyorrhexis, and increase of the apoptotic cells in the combination group, while the promotive effect of low dosage dasatinib on apoptosis of RS4;11 cells was not strong. PCI-32765 and Dasatinib could decrease the expression and activity of BCR-ABL, Btk, Lyn, Src in Sup-B15 and RS4;11 cells.</p><p><b>CONCLUSION</b>PCI-32765 or Dasatinib can inhibit the proliferation and induce the apoptosis of Sup-B15 and RS4;11 cells, PCI-32765 and Dasatinib displayed the synergistic effects. The possible mechanism may be related with the blocking of B cell receptor(BCR) signal pathway, thereby inhibiting the cell proliferation and promoting the cell apoptosis.</p>

Detection of Thrombin Using Double Quenching Molecular Beacon / 分析化学

A double quenching molecular beacon ( MB) with simple structure was designed based on organic quencher and G bases, and a simple detection method for thrombin was developed using this MB. In this MB, FAM and BHQ-1 were selected as fluorophore and organic quencher, three continuous nucleotides with G base were connected with BHQ-1, and the loop of MB was designed as a nucleic acid aptamer of thrombin. In the absence of thrombin, the MB was in the stem-loop structure, the fluorophore FAM was close to BHQ-1 and G bases, the fluorescence of FAM was dual quenched by BHQ-1 and G bases, and the fluorescence signal of FAM was very weak. In the presence of thrombin, MB specifically bound thrombin and formed a G-quadruplex structure. The stem-loop structure of the MB was destroyed, and FAM was separated with BHQ-1 and G bases, leading to recovery of fluorescence of FAM. Under the optimal conditions, the fluorescence intensity of FAM exhibited a good linear relationship with concentration of thrombin in the range of 0. 4-40. 0 nmol/L, and regression equation was △I=24. 63C (nmol/L)+ 13. 06 (R2 =0. 9972) with the detection limit of 0. 18 nmol/L (3σ, n=9). The average recoveries of this method in serum samples were 96. 3%-98. 7%, which indicated that the method had high accuracy.

Detection of Thrombin Using Double Quenching Molecular Beacon / 分析化学

A double quenching molecular beacon ( MB) with simple structure was designed based on organic quencher and G bases, and a simple detection method for thrombin was developed using this MB. In this MB, FAM and BHQ-1 were selected as fluorophore and organic quencher, three continuous nucleotides with G base were connected with BHQ-1, and the loop of MB was designed as a nucleic acid aptamer of thrombin. In the absence of thrombin, the MB was in the stem-loop structure, the fluorophore FAM was close to BHQ-1 and G bases, the fluorescence of FAM was dual quenched by BHQ-1 and G bases, and the fluorescence signal of FAM was very weak. In the presence of thrombin, MB specifically bound thrombin and formed a G-quadruplex structure. The stem-loop structure of the MB was destroyed, and FAM was separated with BHQ-1 and G bases, leading to recovery of fluorescence of FAM. Under the optimal conditions, the fluorescence intensity of FAM exhibited a good linear relationship with concentration of thrombin in the range of 0. 4-40. 0 nmol/L, and regression equation was △I=24. 63C (nmol/L)+ 13. 06 (R2 =0. 9972) with the detection limit of 0. 18 nmol/L (3σ, n=9). The average recoveries of this method in serum samples were 96. 3%-98. 7%, which indicated that the method had high accuracy.

SDF-1α/CXCR4 Mediated Drug Resistance Can be Reversed by Ibrutinib in Acute Lymphoblastic Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of Ibrutinib on the chemoresistance mediated by SDF-1α/CXCR4 axis in ALL cells.</p><p><b>METHODS</b>Flow cytometry was used to detect the apoptosis of cell line and expression of surface membrane CXCR4, Western blot was used to determine the expression level of CXCR4, ERK and Bcl-xL proteins, qPCR was used to assay the mRNA level of CXCR4.</p><p><b>RESULTS</b>Ibrutinib enhanced the apoptosis induced by adriamycin(ADR) (17.100±4.3% to 28.133±3.16%); Ibrutinib inhibited the phosphorylation of CXCR4 induced by SDF-1α and with concentration- and time- dependent manner (r=-0.99659, r=-0.99764, r=-0.99980). Ibrutinib inhibited the expression and activity of CXCR4 downstream signaling molecules pERK and BCL-xL.</p><p><b>CONCLUSION</b>Ibrutinib can enhance the sensitivity of SUP-B15 to ADR, reverse SDF-1α/CXCR4-mediated chemoresistance in Phacute lymphoblastic leukemia cells. This mechanism of ibrutinib may be assosiated with inhibiting CXCR4/ERK/BCL-xL.</p>

Expression of Btk and NFκB in Acute Lymphoblastic Leukemia Cells and Its Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the role of Btk and NFκB in the incidence, development, prognosis and therapeutic efficacy for acute lymphoblastic leukemia(ALL) through detecting their expression in leukemia cells.</p><p><b>METHODS</b>Bone marrow samples from 51 ALL patients were collected, and the mononuclear cells were separated by Ficoll density gradient centrifugation. The expressions of Btk and NFκB at RNA and protein levels were detected by RT-PCR and Western blot respectively.</p><p><b>RESULTS</b>(1)At protein level, Btk and NFκB were expressed in all newly diagnosed 51 ALL patients, among them 38 patients had higher expression level of Btk, 34 patients had high NFκB expression level. The expression of Btk and NFκB was higher in the cells from newly diagnosed ALL patients than that in the cells from patients in CR(P<0.05), and the expression of Btk and NFκB was higher in relapsed ALL patients. (2)The expression of Btk and NFκB in the ALL patients was followed-up higher expression of Btk: among the 38 newly diagnosed B-ALL cases, 27 experienced CR (71%) and 12 of which achieved CR after one course chemotherapy (one course CR) (31%). Moreover, 16 out of the 27 CR patients relapsed after a short period (less than 6 months) (59%). On the contrary, among the 13 patients with low Btk expression, 11 achieved CR (84.6%) after one course and 1 relapsed (8 months after CR) (7.6%). A similar pattern of NFκB expression was observed.</p><p><b>CONCLUSION</b>Btk and NFκ B may play an important role in the incidence and progression of ALL, possibly serving as the potential therapeutic targets of ALL and the indexes for prognosis.</p>

Influence of Leukodeplated Blood Transfusion on Cellular Immunity of Acute Leukemia Patients / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the influence of leukodeplated blood transfusion on cellular immunity of patients with acute leuemia, so as to provide support for application of leuko-deplated blood transfusion in clinic.</p><p><b>METHODS</b>A total of 100 AL patients from January 2012 to December 2015 were chosen, and were divided into 2 groups: leukodeplated blood transfusion group(50 cases) and routine blood transfusion group(RBT) as control (50 cases). The effective rate, side effects, peripheral blood T cells and expression level of TLR2 and TLR4 were compared between 2 groups.</p><p><b>RESULTS</b>The expression levels CD3(+), CD4(+), CD8(+), CD4(+)/CD8(+) of TLR2 and TLR4 in control group were (52.18±2.14)%, （27.28±1.19）%,（24.21±1.65）%,1.22±0.18,0.62±0.04 and 0.57±0.05, respectively, after treatment; while these indicators in LdBT group were （52.18±2.14）%,（30.97±2.01）%,（27.08±1.55）%,1.39±0.24,0.91±0.06 and 0.87±0.07, respectively, and above-mentioned indicators in LdBT group were significantly higher than those in control group（P<0.05）. Compared with these indicators before treatment, CD3(+), CD4(+), CD8(+) and CD4(+)/CD8(+) in the patients increased significantly（P<0.05）. The efficiency was 92.00% (46/50) in LdBT group, and 84.00% (42/50) in control group, without statistically significant difference（P>0.05）. The rate of side effects in study group was 6% (3/50), 18% (9/50) in control group, with statistically significance difference （P<0.05）.</p><p><b>CONCLUSION</b>Leukodeplated blood transfusion can improve the cellular immunity of AL patients, and reduce the rate of side effects.</p>

Effects of Salvianolate on Myosin Heavy Chain in Cardiomyocytes of Congestive Heart Failure Rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of Salvianolate on myosin heavy chain (MHC) in cardiomyocytes of congestive heart failure (CHF) rats.</p><p><b>METHODS</b>Sixty male SD rats were divided into 6 groups according to random digit table, i.e., the normal control group (NCG), the model group, the Captopril group (CAG), the low dose Salvianolate group (LSG), the high dose Salvianolate group (HSG), the Captopril and high dose Salvianolate group (CSG), 10 in each group. CHF rat model was established with peritoneal injection of adriamycin in all rats except those in the NCG. Equal volume of normal saline was peritoneally injected to rats in the NCG, once per week for 6 successive weeks. Corresponding medication was started from the 5th week of injecting adriamycin. Rats in the CAG were administered with Captopril solution at the daily dose of 10 mg/kg by gastrogavage. Rats in the LSG and the HSG were administered with Salvianolate solution at the daily dose of 24.219 mg/kg and 48.438 mg/kg respectively by gastrogavage. Salvianolate was dissolved in 2 mL 5% glucose solution and administered by peritoneal injection. Rats in the CSG were peritoneally injected with high dose Salvianolate solution and administered with Captopril solution by gastrogavage. Two mL normal saline was peritoneally injected to rats in the model group, once per day for 8 successive weeks. Eight weeks later, the cardiac function and myocardial hypertrophy indices were detected by biological signal collecting and processing system. mRNA expression levels of alpha-MHC and beta-MHC in cardiac muscle were detected by fluorescence quantitative PCR. Expressions of protein kinase C (PKC) in cardiac muscle were detected by Western blot.</p><p><b>RESULTS</b>Compared with the normal control group, heart mass index (HMI) and left ventricular mass index (LVMI) obviously increased in the model group (P < 0.01). Compared with the model group, HMI and LVMI decreased in HSG, CAG, and CSG groups (P < 0.05, P < 0.01). It was more obviously lowered in the CSG group than in the CAG group (P < 0.05). Compared with the NCG, the mRNA expression level of alpha-MHC in cardiac muscle decreased, the mRNA expression level of p-MHC and the expression of PKC in cardiac muscle increased in the model group (P < 0.01). Compared with the model group, the mRNA expression level of alpha-MHC in cardiac muscle was increased, and the mRNA expression level of beta-MHC and the expression of PKC in cardiac muscle were decreased in HSG, CAG, and CSG groups (P < 0.05, P < 0.01). There was statistical difference between the CSG group and the CAG group (P < 0.05).</p><p><b>CONCLUSIONS</b>Salvianolate could up-regulate the mRNA expression level of alpha-MHC, and down-regulate the mRNA expression level of beta-MHC in cardiac muscle. Its mechanism might be related to decreasing the expression of PKC.</p>