[Clinical Characteristics and Prognostic Factors of Primary FL Patients with Grade 3 or Large B Cell Transformation].

OBJECTIVE: To investigate the clinical characteristics and prognostic factors of primary follicular lymphoma (FL) patients with grade 3 or large B cell transformation, so as to provide more reference for the subsequent clinical diagnosis and treatment. METHODS: Forty-seven primary FL patients with grade 3 or large B cell transformation from March 2010 to March 2018 were selected, the clinical characteristics and survival of patients were analyzed. Cox regression model were used to evaluate the related prognostic factors. RESULTS: The cumulative progression-free survival rate and cumulative overall survival rate of 47 patients in 3-year follow-up reached to 55.32% (26/47) and 80.85% (38/47) respectively. There were significant differences in cumulative progression-free survival rate and cumulative overall survival rate among different subgroups of IPI, FLIPI-1 and FLIPI-2 in 3-year follow-up (P<0.05). The cumulative progression-free survival rate during follow-up for 3-year in the patients with Ann Arbor staging for III-IV stage, lymph node-involved site≥5, lymph node-involved site with diameter more than 3 cm≥3 and extranodal lesions≥3 was significantly lower than other subgroups (P<0.05). The cumulative overall survival rate in 3-year follow-up of patients with LDH levels≥240 U/ml was significantly lower than patients with LDH levels < 240 U/ml (P<0.05). Univariate analysis showed that Ann Arbor stage for III-IV, lymph node-involved site number≥5, >3 cm lymph node-involved site number≥3, extranodal lesion site number≥2, IPI score=2-3, FLIPI-1 score and FLIPI-2 score≥3 were the risk factors for progression-free survival (P<0.05); LDH≥240 U/ml, IPI score=2-3 and FLIPI-2 score≥3 were risk factors for overall survival (P<0.05). Cox regression model multivariate analysis showed that IPI score=2-3 was the independent risk factor for progression-free survival and overall survival (P<0.05). FLIPI-2 score≥3 was the independent risk factor for overall survival (P<0.05). CONCLUSION: Primary FL patients with grade 3 or large B cell transformation by using the existing treatment regimen might be possibly curable, and the current treatment strategies and IPI score can be used to predict the clinical prognosis of patients.

[Effect of MiRNA-145 on Apoptosis of Leukemia HuT 78 Cells and Its Mechanism].

OBJECTIVE: To investigate the effect and mechanism of miRNA-145 on leukemic cell apoptosis. METHODS: After transfection of miRNA-145 mimic and negative control mimic in leukemia cells by Lipofectamine 2000 liposome, the MTT assay was used to detect the effect of miRNA-145 on cell proliferation. Flow cytometry was used to detect the effect of miRNA-145 on cell cycle and apoptosis. Western blotting assay was used to detect the expression levels of BCL-2, CDK6, Cyclin D1, BAX, PI3K p-PI3K, p-AKT and AKT. RESULTS: The relative level of microRNA in HuT 78 cells transfected with miRNA-145 was 2.3±02, which was significantly higher than that in blank control group and miRNA-NC group (P＜0.05). MTT assay showed that the proliferation level of HuT 78 cells transfected with miRNA-145 mimic was significantly lower than that of blank control and miRNA-NC group (P＜0.05). Flow cytometry showed that the cells at G0/G1, S and G2/M phase of HuT 78 cells were significantly decreased after transfection with miRNA-145 mimic (P＜0.05). Annexin V/PI double staining assay showed that the apoptosis rate of HuT 78 cells was 17.6%±3.4%，which was significantly higher than that in blank control group and miRNA-NC group (P＜0.05). Western blot showed that the expression levels of BCL-2, CDK6 and Cyclin D1 in HuT 78 cells were significantly lower than those in blank control and miRNA-NC group (P＜0.05), and BAX expression in HuT 78 cells was significantly higher than that in blank control and miRNA-NC group (P＜0.05). Western blot showed that expression of PI3K, p-PI3K, AKT and p-AKT in HuT 78 cells transfected with miRNA-145 mimic were significantly lower than that in blank control and miRNA-NC group (P＜0.05). CONCLUSION: Upregulation of miRNA-145 may inhibit the proliferation of leukemia cells and promote the apoptosis, which may be related with the inhibition of PI3K/AKT signaling pathway.

HLA-Mismatched Microtransplant in Older Patients Newly Diagnosed With Acute Myeloid Leukemia: Results From the Microtransplantation Interest Group.

IMPORTANCE: The outcome of older patients with acute myeloid leukemia (AML) remains unsatisfactory. Recent studies have shown that HLA-mismatched microtransplant could improve outcomes in such patients. OBJECTIVE: To evaluate outcomes in different age groups among older patients with newly diagnosed AML who receive HLA-mismatched microtransplant. DESIGN, SETTING, AND PARTICIPANTS: This multicenter clinical study included 185 patients with de novo AML at 12 centers in China, the United States, and Spain in the Microtransplantation Interest Group. Patients were divided into the following 4 age groups: 60 to 64 years, 65 to 69 years, 70 to 74 years, and 75 to 85 years. The study period was May 1, 2006, to July 31, 2015. EXPOSURES: Induction chemotherapy and postremission therapy with cytarabine hydrochloride with or without anthracycline, followed by highly HLA-mismatched related or fully mismatched unrelated donor cell infusion. No graft-vs-host disease prophylaxis was used. MAIN OUTCOMES AND MEASURES: The primary end point of the study was to evaluate the complete remission rates, leukemia-free survival, and overall survival in different age groups. Additional end points of the study included hematopoietic recovery, graft-vs-host disease, relapse rate, nonrelapse mortality, and other treatment-related toxicities. RESULTS: Among 185 patients, the median age was 67 years (range, 60-85 years), and 75 (40.5%) were female. The denominators in adjusted percentages in overall survival, leukemia-free survival, relapse, and nonrelapse mortality are not the sample proportions of observations. The overall complete remission rate was not significantly different among the 4 age groups (75.4% [52 of 69], 70.2% [33 of 47], 79.1% [34 of 43], and 73.1% [19 of 26). The 1-year overall survival rates were 87.7%, 85.8%, and 77.8% in the first 3 age groups, which were much higher than the rate in the fourth age group (51.7%) (P = .004, P = .008, and P = .04, respectively). The 2-year overall survival rates were 63.7% and 66.8% in the first 2 age groups, which were higher than the rates in the last 2 age groups (34.2% and 14.8%) (P = .02, P = .03, P < .001, and P < .001, respectively). The 1-year cumulative incidences of nonrelapse mortality were 10.2%, 0%, 3.4%, and 26.0% in the 4 age groups and 8.1% in all patients. The median times to neutrophil and platelet recovery were 12 days and 14 days after induction chemotherapy, respectively. Five patients had full or mixed donor engraftment, and 30.8% (8 of 26) of patients demonstrated donor microchimerism. Two patients (1.1%) developed severe acute graft-vs-host disease. CONCLUSIONS AND RELEVANCE: Microtransplant achieved a high complete remission rate in AML patients aged 60 to 85 years and higher 1-year overall survival in those aged 60 to 74 years.

[Analysis of Indicators Related with Cell Proliferation and Apoptosis in the Patients with B Cell Non-Hodgkin's Lymphoma].

OBJECTIVE: To analyze the indicators related with cell proliferation and apoptosis in patients with B cell non-Hodgkin's lymphoma(B-NHL). METHODS: Seventy-two cases of B-NHL and 50 cases of reactive hyperplasia of lymph nodes were entrolled in the experimental group and control group respectively. The expression levels of proliferating cell nuclear antigen(PCNA), X-linked inhibitor of apoptosis(XIAP) and B cell lymphoma leukemia-2(BCL-2) in paraffin-embedded tissue samples of 2 groups were detected by immuno-histochemistry staining, and their ralationship with pathologic factors was analyzed. RESULTS: The positive cell rates of PCNA, XIAP and BCL-2 in experiment group were significantly higher than those in control group (P<0.05); the positive cell rate of PCNA in B-NHL patients at III-IV stage was higher than that in B-NHL patients at I-II stage(P<0.05), however, the positive cell rates of XIAP and BCL-2 in B-NHL patients with different pathologic factors were not significantly different(P>0.05). The progression-free survival(PES) time in PCNA low positive expression group was longer than that in PCNA high positive expression group(P<0.05), but the PFS time between B-NHL patients with XIAP positive and negative expression was not significantly different(P>0.05); the PFS time also was not significantly different between B-NHL patients with BCL-2 positive and negative expression(P>0.05). CONCLUSION: All the PCNA, XIAP and BCL-2 participate in the pathngenesis of B-NHL, among them the positive level of PCNA obviously influences the clinical staging of B-NHL.

[Effect of G Protein-coupled Receptor Kinase 6 on Proliferation and Apoptosis of Multiple Myeloma Cells and Its Mechanisms].

OBJECTIVE: To study the effect of G protein-coupled receptor kinase 6(GRK6) on proliferation and apoptosis of multiple myeloma(MM) cells and its mechanisms. METHODS: The samples were collected from MM patients and healthy people for study in vivo. The plasma cells isolated from multiple myeloma patients, as well as U266 and NCI H929 myeloma cell lines were used for study in vitro. Western blot and quantitative real-time PCR were used to evaluate the protein and mRNA of expression of GRK6 in multiple myeloma, cell proliferation and apoptosis were tested by BrdU and Annexin V-FITC/PI assays. RESULTS: The protein and mRNA expression of GRK6 in multiple myeloma was higher than those in control group, and the expression level of GRK6 in stage I of MM was higher than that in control group, while the expression level of GRK6 in stage II was higher than that in stage I, but lower than that in stage III (P<0.05). U266 and MM cells showed high-sensitivity to CX-4945, except NCI H929. GRK6 expression level in U266, NCI H929 and MM cells treated with siRNA and CX-4945, significantly decreased as compared with those cells treated by CX-4945 alone. Cell proliferations of U266, NCI H92 and MM groups treated with CX-4945 were (58.25±18.24)%, (64.32±20.03)% and (45.42±25.01)% respectively, moreover, their apoptotic rates were (62.82±53.21)%, (43.25±47.05)% and (85.67±40.32)% respectively. CONCLUSION: The expression level of GRK6 in multiple myeloma increases, and GRK6 inhibitor CX-4945 inhibits proliferation and promotes apoptosis of myeloma cells; GRK6 regulates Rac1 and involves in the proliferation and apoptosis pathway of multiple myeloma cells.

[Infection Status and Prognosis Analysis of Epstein-Barr Virus in Patients with Newly Diagnosed Hodgkin's Lymphoma].

OBJECTIVE: To investigate the Epstein Barr virus(EBV) positive rate in newly diagnosed Hodgkin's lymphoma(HL) and the prognostic significance of EBV status. METHODS: A total of 120 previously untreated patients with histologically confirmed Hodgkin's lymphoma were enrolled in this study. The EBV infection status was confirmed through examining EBV-RNA(EBER) or EBV latent membrane protein-1, and these patients were divided into EBV positive group and EBV negative group. The correlation of clinical features and EBV infection status was analyzed. For analysis of prognostic significance of EBV infection, the patients were divided into dead and survival groups, and the factors affecting the living conditions were analyzed. RESULTS: Among 120 patients with HL, 36 patients(30.0%) were found with EBV infection. In EBV+ HL group patients were male, aged 6-15 and 61-74, the proportion of patients with mixed cellular sybtype was significantly higher than that in EBV- HL group(P<0.05). The 1 year and 2 year total survival rate of patients in EBV+ group were 88.9% and 83.3%, and significantly lower than 97.6% and 95.2% in EBV- group. Out of the 120 patients with HL, 10 patients died(8.3%). In death group, patients aged 61-74 and did not received radiotherapy, their proportion of EBV+ infection was significantly hyher than that in survival group (P<0.05). Multiriabl analysis showed that the age 61-74 and EBV+ infection were the risk factors for survival coditions of patients (P<0.05). CONCLUSION: The EBV infection relates with HL, the clinical featuses of HL patients with EBV+ or EBV- are different, the total survival time of patients in EBV+ group is shorter than that of patients in EBV- group, the EBV+ infection is a risk factor for total survival time of patients with HL.

[Clinical Efficiency and Safety of the First-line CHOP Regimen Containing PLD Applied to Treat Aged Patients with Advanced DLBCL].

OBJECTIVE: To explore the clinical efficiency and safety of CHOP regimen containing pegylated liposomal doxorubicin (PLD) for the aged patients with advanced diffuse large B-cell lymphoma (DLBCL). METHODS: Fifty aged patients with advanced DLBCL treated in our hospital from February 2010 to February 2014 were selected and divided into two groups. Out of 50 cases, 25 cases received standard CHOP regimen (sCHOP group), other 25 cases received CHOP regimen containing PLD at dose of 30 mg/m2 (PLD+CHOP). These patients were followed up for 18 months, and the total effective rate, the survival rate and the adverse reaction rate were compared between these two groups. RESULTS: After receiving different treatments, the survival rate of patients on 6, 12 and 18 months in PLD+CHOP group was 88.0%, 80.0% and 76.0%, respectively, and the survival rate of 18 month was significantly higher than that in the sCHOP group (P<0.05); The total effective rate in the PLD+CHOP group was statistically higher than that in the sCHOP group (P<0.05); and all the incidences of non-hematological toxicity, peripheral sensory neuropathy, lung infection, gastrointestinal reaction and hepatotoxicity were not statistically different between two groups (P>0.05), while the incidence of cardiac toxicity including acute myocardial infarction, congestive heart failure, atrioventricular block (AV block) and paroxysmal atrial tachycardia significantly decreased in the PLD+CHOP group (P<0.05). CONCLUSION: The efficiency of CHOP regimen containing PLD for the aged patients with advanced DLBCL has been confirmed to be significant, and its cardiac toxicity is low, thus being worth to be popularized and applied for the treatment of advanced diffuse large B-cell lymphoma.

[Clinical Comparative Study of Two Kind Doses of Bortezomib Combinated with Bisphosphonates for Treating Patients with Multiple Myeloma Ostespathy].

OBJECTIVE: To investigate the clinical efficacy and safety of conventional dose and reduction dose of bortezomib in combination with bisphosphonates for treating patients with multiple myeloma ostespathy. METHODS: A total of 150 patients with multiple myeloma ostespathy were chosen in the period from March 2011 to July 2015 and randomly were divided into 2 groups: A group (75 cases) and B group (75 cases). The patients in A and B groups were treated with conventional dose of bortezomib and reduction dose of bortezomib on the basis of bisphosphonates respectively and the clinical efficacy, the improvement rate of life quality, NRS score, levels of IL-6 and CRP before and after treatment, and the adverse effects of 2 groups were compared. RESULTS: There was no significant difference in the clinical efficacy between 2 groups (P<0.05). The improvement rate of patients life quality in B group was significantly better than that in A group (P>0.05). There was no significant difference in the NRS score, levels of IL-6 and CRP after treatment between 2 groups (P>0.05). There was no significant difference in the incidence of neutrophil reduction and thrombocytopenia between 2 groups (P<0.05). The incidence of BiPN, nausea and vomiting, herpes zoster and fatigue of B group was significantly lower than that in A group (P<0.05). CONCLUSION: Conventional dose and reduction dose of bortezomib in combination with bisphosphonates for treating patients with multiple myeloma ostespathy possess the same effects, including pain relief and disease progression control; but the reduction dose of bortezomib application can efficiently improve the life quality of patients and reduce the risk of adverse reactions.

MiR-301a promotes cell proliferation by directly targeting TIMP2 in multiple myeloma.

BACKGROUND: Multiple myeloma (MM) is a plasma cell malignancy characterized by clonal proliferation of plasma cells in the bone marrow and microRNAs play a crucial role in its tumorigenesis and development. The purpose of this study was to investigate the biological functions of miR-301a in MM. METHODS: Quantitative real-time PCR was used to detect the expression level of miR-301a. Cell proliferation was assessed by MTT assay. Flow cytometry was performed to valuate cell apoptosis and cell cycle distribution. Moreover, luciferase reporter assay and western blot were conducted to determine the potential target of miR-301a in MM cells. RESULTS: MiR-301a is significantly up-regulated in MM clinical bone marrow samples and cell lines compared with normal controls. Gain-of-function and loss-of-function studies in MM cell line U266 showed that miR-301a acts as an oncogene in MM by promoting cell proliferation and inhibiting apoptosis. Furthermore, a tumor suppressor gene, tissue inhibitor of metallopeptidases-2 (TIMP2) was identified as a direct target of miR-301a and knockdown of TIMP2 could mimic the effect of miR-301a in MM. CONCLUSIONS: MiR-301a promotes cell proliferation and inhibits apoptosis by direct targeting TIMP2 in MM, and miR-301a might represent a novel molecular in MM and may provide helpful therapeutic strategies for MM treatment.

MicroRNA-204 inhibits cell proliferation in T-cell acute lymphoblastic leukemia by down-regulating SOX4.

BACKGROUND: MicroRNAs (miRNAs) are a group of small non-coding RNAs that play important roles in the pathogenesis of human diseases by negatively regulating gene expression. The aim of this study was to explore the effect of miR-204 on cell proliferation migration and invasion in T-cell acute lymphoblastic leukaemia (T-ALL). METHOD: miR-204 expression was determined in bone marrow samples from 32 leukemia patients and 32 healthy controls by quantitative real-time PCR (qRT-PCR). The effect of miR-204 on cell proliferation was evaluated by CCK8 assay, cell migration and invasion were evaluated by transwell migration and invasion assays, In addition, the regulation of SOX4 by miR-204 was evaluated by luciferase reporter assay and western blot. RESULTS: our results revealed that miR-204 was low expressed in T-ALL. Cell proliferation assay showed that the cell proliferation ability was inhibited by miR-204 mimics. Moreover, migration and invasion assay suggested that overexpression of miR-204 could significantly suppressed the migration and invasion ability of T-ALL cells. Luciferase reporter assay confirmed that miR-204 directly bound to the 3' untranslated region of SOX4, and western blot suggested that miR-204 inhibited the expression of SOX4 at the protein levels. CONCLUSIONS: Our findings indicated that miR-204 negatively regulates SOX4 and inhibited proliferation, migration and invasion of T-ALL cell lines. Thus, miR-204 might represent a potential therapeutic target for T-ALL intervention.