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Objective: To investigate the relationship between clinical indicators of CRAB symptoms and antioxidant enzyme activity in patients with multiple myeloma (MM)., Methods: The activity of catalase (CAT), glutathione peroxidase (GPX), and superoxide dismutase (SOD) in the bone marrow supernatants of 44 patients with MM and 12 patients with non-malignant hematological diseases was detected by colorimetric assay, and then the differences in the activity of antioxidant enzymes between the two groups were compared. Furthermore, the relationship between the activity of antioxidant enzymes in the MM group and the levels of serum calcium, serum creatinine (Scr), hemoglobin (Hb), alkaline phosphatase (ALP) as well as bone lesions were analyzed., Results: The antioxidant enzyme activity was lower in MM patients compared with the control group ( P < 0.05). When the concentrations of serum calcium and ALP were higher than the normal levels, Hb was lower than 85 g/L, and there were multiple bone lesions, the activity of CAT, SOD and GPX was significantly declined ( P < 0.05); When the concentration of Scr≥177 μmol/L, the activity of GPX was significantly declined ( P < 0.05). Regression analyses showed that CAT, SOD and GPX were negatively correlated with serum calcium ( r =-0.538, r =-0.456, r =-0.431), Scr ( r =-0.342, r =-0.384, r =-0.463), and ALP ( r =-0.551, r =-0.572, r =-0.482)., Conclusion: The activity of antioxidant enzymes, including CAT, SOD and GPX, were decreased in patients with MM and they were negatively correlated with some clinical indicators of CRAB symptoms (such as serum calcium, Scr, and ALP), which suggests that promoting the activity of antioxidant enzymes may be beneficial to treat the CRAB symptoms of the patients with MM.

Objective: To investigate the efficacy, safety and prognosis of auto-HSCT between classical and modified conditioning regimen in patients with B-cell non-Hodgkin lymphoma., Methods: 36 patients diagnosed as B-cell non-Hodgkin lymphoma treated with autologous hematopoietic stem cell transplantation from January 2015 to June 2018 in Tianjin Cancer Hospital were retrospectively analyzed. The patients were divided into two groups: Idarubicin group and non-Idarubicin group. The overall survival (OS), progression-free survival (PFS), adverse reactions and hematopoietic reconstitution time between the two groups were compared. Survival analysis was performed by using the Kaplan-Meier method. Log-rank test was used for comparison between groups, and Cox regression was used for multivariate analysis., Results: The median follow-up time was 29 months. Among these 36 patients with B-cell non-Hodgkin lymphoma before transplantation, 21 patients achieved CR and 15 patients achieved PR. The reconstitution time of neutrophil (P>0.05) and platelet (P>0.05) was not significantly different between Idarubicin and non-Idarubicin group. Also, the adverse reactions were not significantly different between two groups. The addition of idarubicin showed not aggravate the adverse reactions of patients. The OS and PFS of patients with idarubicin was longer than that of patients without idarubicin. The multivariate analysis showed that, the modified conditioning regimen and the remission state before transplantation were closely associated with prognosis., Conclusion: The above-mentioned results indicated that the combination of modified conditioning regimen with idarubicin can lengthen the OS and PFS of the patients significantly, and show not aggravate of bone marrow inhibition, moreover, the hematopoietic reconsititution time show not lengthen, which means that it can be a safe and effective choice for autologous HSCT in the patients with B cell non-Hodgkin lymphoma.

Exosomes are microvesicles that can be secreting in many kinds of cells, under the condition of normal and pathology. Exosomes contain abundant proteins, miRNAs and RNA fragments, which play an important role in communicating between cells, especially non-contact cells. The interaction between blood tumor cells and immune microenvironment is inseparable from the involvement of exosomes. Exosomes are involved in the interaction among tumor cells, NK cells, T cells, stromal cells and endothelial cells, such as promoting the proliferation of blood tumor cells, helping blood tumor cells to achieve immune evasion, promoting angiogenesis and migration of tumor cells. Therefore, exosomes are closely related to the diagnosis, treatment and prognosis of hematological tumors. In this review, the basic characteristics of exosomes, the important role of exosomes in the immune microenvironment of hematological tumors, and the research progress of exosomes in clinical applications was sammrized briefly.

Objective: To analyze the clinical and pathological characteristics of primary gastrointestinal non-Hodgkin's lymphoma (PGI-NHL) patients, and to explore the factors affecting the patients' survival and prognosis., Methods: The clinical data of 219 patients with PGI-NHL diagnosed in our hospital from March 2009 to April 2016 was collected and retrospectively analyzed. Survival analysis was performed by using the Kaplan-Meier method. Log-rank test was used for comparison among the groups, and Cox regression was used for multivariate analysis., Results: Among the 219 patients with PGI-NHL, 126 patients were males and 93 patients were females. 182 patients were IPI 0 to 2 and 37 patients were IPI 3 to 5. There were 205 cases (93.6%) of B cell phenotype and 14 cases (6.4%) of T cell phenotype. 140 patients (63.9%) were patients with primary gastric NHL, including 85 DLBCL and 19 MALT. 79 cases (36.1%) were patients with primary intestinal NHL, including 46 DLBCL, 4 MALT, 7 FL, 3 MCL and 4 Burkitt lymphoma. 23 cases were HP positive and received anti-HP therapy. 57 cases and 32 cases received surgery and chemotherapy respectively. 84 cases received combination treatment of surgery and chemotherapy and 11 cases received combination treatment of radiotherapy and chemotherapy. Overall survival (OS) of indolent B-cell non-Hodgkin's lymphoma was longer than that of invasive B-cell non-Hodgkin's lymphoma, which shows better prognose. Kaplan-Meier analysis showed that there was no difference between progression-free survival (PFS) and OS in the patients with different origin sites, age and sex. There was no significant difference in PFS between B-cell and T-cell-derived patients, whereas OS of B-cell-derived PGI-NHL patients was longer than that of T-cell-derived PGI-NHL patients. The OS and PFS of patients with IPI 0-2 were longer than those of patients with IPI 3-5. According to Lugano and Ann Arbor staging systems, there was no difference in prognosis of patients between phase I/II and III/IV. The prognosis of patients treated with surgery alone was worse than that of patients treated with combination therapy, and the prognosis of patients with surgery combined with chemotherapy was not significantly different from that of patients with chemotherapy alone., Conclusion: B-cell phenotype, indolent and low IPI score lymphoma indicate better prognosis, while that of different origin site, sex and age shows no different in prognosis. Surgery is used only for emergency case or pathological materials, and these patients should be treated with chemotherapy-based combined treatment.

Objective: To compare the prognostic value of different staging systems in primary intestinal diffuse large B cell lymphoma（PI-DLPCL）, and their correlation with clinicopathological characteristics，treatment and prognosis of PI-DLBCL., Methods: A total of 68 patients with PI-DLBCL were recruited from January 2009 to July 2017. All the patients underwent staging by using TNM, Lugano, Blackledge and Musshoff system, survival curves for the PI-DLBCL patients were plotted using the Kaplan-Meier method and were judged by the log-rank test. The accuracy of each staging system for predicting survival of PI-DLBCL patients was evaluated by calculating the area under curve(AUC) of the receiver operating characteristic(ROC). The correlation of the 4 staging systems, clinical features patients and treatment regimes with PFS and OS were analysed., Results: The median follow-up time was 52 (1-105) months, the median PFS time was 41(1-86) months, patients did not reached the median OS time. The most frequently involved site was ileocecal (30.9%), followed by small intestine (29.4%) and colon (29.4%), multiple sites involvement (7.4%) and rectum (2.94%)．The PFS and OS rates at 5-year were 44.9% and 51.1%, respectively. Kaplan-Meier survival curves and log-rank test results showed that using different staging systems to describe the cumulative retention rates of PFS and OS in PI-DLBCL patients, none of the 4 staging systems can distinguish the survival curves of each stage significantly. The results of ROC curve showed that the prediction ability of the Lugano staging system was better than other staging system for 1 year PFS （AUC=0.826；P=0.015）and 1 year OS（AUC=0-792；P=0.001） in PI-DLBCL patients. The 3 year PFS rate in the operation+chemo or radio-therapy group (62 cases) and the single operation group (6 cases) were 53.9% and 16. 7%,respectively（P=0.116），The 3 year OS rate were 66.7% and 16.7%（P=0.015）,respectively. Patients who received chemotherapy combined with rituximab had a higher 3-year PFS(66.0％ vs 44.0％，P=0.139) and 3.year OS(70.2% vs.39.2%，P=0.148).The patients with ileocecal lesion had higher PFS rate and OS rate than other sites(P<0.05). Multivariate Cox regression analysis indicated that only bone marrow invasion was an independent prognostic factor in patients with PFS., Conclusion: Bone marrow invasion is an independent risk factor for PFS in patients with PI-DLBCL , according to this limited preliminary data，Lugano staging system for stratifying and predicting the prognosis of PI-DLBCL patients is better than other staging system.

Objective: To evaluate the quality of life (QOL) on patients with multiple myeloma(MM) during maintenance therapy and to explore the related factors important for QOL., Methods: The demography, clinical and laboratorial data of 66 MM patients during maintenance therapy were collected and explored by using a cross-sectional question naire(EORTC QLQ C30 V 3.0). The statistical analysis was performed using Nowegram normal mode(NM) and reference values(RV) of MM patients which were used as control., Results: In comparison with Nowegran normal mode, the scores of general health status, physical function, role function and social function of patients during maintenance therapy were lower than those of normal mode (61.3, 73.9, 65.4 and 65.2 vs 75.3, 89.9, 83.3 and 85.8 respectively), while the scores of constipation and financial difficulty were higher than those of normal mode(16.7 and 44.4 vs 10.7 and 9.7 respectively) (P<0.05). In comparison with reference values, the scores of general health status, emotional and coguitive functions of patients during maintenance therapy were significantly higher than those of reference values(61.3, 81.7 and 84.3 vs 55.7, 71.3 and 78.1 respectively) (P<0.05). In addition, the maintenance therapy yet decreasd the scores of fatigue, nausea and vomiting, pain, dyspnoea, insomnia, appetite loss and constipation of patients, but increased the score of financial difficulty of patients (P<0.05). The age of initial diagnosis, serum LDH level, peripheral neuropathy, high ratio of own expense and underlying diseases were main factors affecting the general health status of patients (P<0.05), while the decrease of Hb level, increase of blood Ca ++ level and accompanied genetic changes negatively influence the QOL (P<0.05), while the high culture level showed positive effect on QOL (P<0.05). The choise of drugs for maintenace (therapy thalidomide and bortezomib) not had significant effect on QOL of patients., Conclusion: The maintenance therapy can improve the QOL of MM patients, the age at initial diagnses, serum LDH level, peripheral neuropathy and high ratio of own expence are the main factors affecting the QOL of MM patients.

Objective: To explore the maintaining measures for the vitality of hematopoietic stem cells (HSC) in vitro, so as provide technical support for ultra long distance transport of HSC collected from unrelated donors., Methods: Peripheral blood hematopoietic stem cells (PBHSC) were treated by different methods according to various groups, then stored at 4 ℃ in the refrigerator. The percentage of CD34 + cells, relative cell activity, relative cell proliferation rate, relative colony-forming rate, oxygen fraction and intracellular reactive oxygen species (ROS) were detected at 0, 24, 48 and 72 h after storage of PBHSC respectively., Results: The percentage of CD34 + cells during 72 h storage did not altered. Along with the prolonging of storage time, the relative cell activity, relative cell proliferation rate and relative colony-forming rate gradually decreased in untreated PBHSC(control group), the related coefficients were -0.796, -0.883 and -0.815 respectively. Plasma dilution, antioxidants and oxygenation could improve the relative cell activity and relative cell proliferation rate, but oxygenation could decrease the relative colony-forming rate of PBHSC. The combination of 2 or 3 factors showed stronger protection effects on PBHSC. The intracellular level of ROS decreased gradually with the prolonging of storage time. Oxygenation of PBHSC could increase oxygen fraction, and also increase the intracellular level of ROS at the same time. The addition of antioxidants could reduce the level of ROS., Conclusion: The percentage of CD34 + cells can not serve as the indicator of PBHSC vitality. Plasma dilution, oxygenation and antioxidants can increase the survival and viability of PBHSC, but oxygenation can increase the intracellular ROS level and impair colony-forming ability of PBHSC. The combination of multiple factors can maintain the vitality of PBHSC better.

Diffuse large B cell lymphoma（DLBCL）is the most common non-Hodgkin's lymphoma in adults，accounting for 30%-40% of non-Hodgkin's lymphoma in adults. The treatments of this lymphoma mainly include chemotherapy，radiotherapy and autologous hematopoietic stem cell transplantation. A large amount of studies show that autologous hematopoietic stem cell transplantation considered as the main treatmant approach can be used for primary high-risk young and relapsed or refractory DLBCL patients. Many scholars have evaluated the effectiveness of autologous hematopoietic stem cell transplantation for primarily high-risk young and relapsed or refractory DLBCL patients，and tried to improve the effectiveness of transplantation that showed some advantagcs for DLBCL patients.

Objective: To evaluate the therapeutic effect and adverse reactions of the maintenance therapies with Thalidomine or Bortezomib in the patients with newly diagnosed multiple myeloma (MM), so as to provide a reference for clinical treatment., Methods: A retrospective analysis was conducted to compare the progression-free survival (PFS), overall survival (OS) and adverse reaction rate of 23 MM patients received the maintenance therapies of Bortezomib and of 68 MM patients received maintenance therapy of Thalidomine., Results: The maintenance therapy with Bortezomib could extend the PFS of MM patients as compared with Thalidomine (PFS rate of patients on the maintenance therapy of Bortezomib in 12th, and 24th month was 100%, 88.89%, and that of Thalidomine-treated group was 72.31%, 47.54%). What's more, some specific patients could get better 2-year PFS rate in Bortezomib group than that in Thalidomine group, such as older than 65 years old, after autologous hematopoietic stem cell transplantation(ASCT), having genetic changes, extramedullary lesions, poor renal function, low serum free light chain ratio, high β2-MG, anemia, high LDH, VGPR of induction and consolidation therapy. The OS rate of Bortezomib on 18th, 24th and 30th month was 100%, 88.89%, 80% verus 91.52%,83.63%,72.90% of the group with thalidemide at the same time. As for 2-year OS rate, the Bortezomib group was higher than Thalidomine without statistical differences. However, the patients such as older than 65 years old, poor renal function and with extramedullary lesions, would also get higher 2-year OS rate from Bortezomi. Bortezomib and thalidomide could cause bone marrow suppression, peripheral neuritis and other adverse reactions., Conclusion: The efficacy of maintenance therapy with Bortezomib is superior to thalidomide. As a conclusion, bortezomib is a better option for maintenance therapy of MM patient.

Objective: To evaluate the therapeutic efficacy and prognosis of autologous stem Hematopoietic cell transplantation (auto-HSCT) in multiple myeloma (MM) patients., Methods: A retrospective study was conducted for 56 patients diagnosed with MM and then received auto-HSCT in our hospital from December 2008 to September 2016., Results: All the patients successfully underwent hematopoietic reconstruction without transplantation-related mortality (TRM). The complete response (CR) rate of all the patients after induction chemotherapy was 23.2% (13/56), while the CR rate of these patients with auto-HSCT increased to 78.6% (44/56) (P<0.01). The CR plus VGPR (very good partial response) rates of these 56 patients after induction chemotherapy and auto-HSCT were 53.6%（30/56）and 94.6%（53/56） respectively (P<0.01). The median progression-free survival (PFS) time and median overall survival (OS) time were 37 and 71 months, respectively. The median PFS time in the patients with induction therapy containing bortezomib was 37 months, however, the median OS time did not reach to 71 months; the median PFS (P<0.01) and the median OS (P<0.01) in the patients with the induction chemotherapy without bortezomib was 27 and 51 months, respectively. Univariate analysis demonstrated that the patients maintained CR or VGPR after auto-HSCT or with less than 6 cycles of induction chemotherapy significantly correlated with PFS (P<0.01)., Conclusion: auto-HSCT can further increase the CR rate, prolong PFS and OS time. Sequential auto-HSCT after bortezomib-based therapy is the first line therapy for the transplant-eligible MM patients. Maintenance treatment is beneficial to the sustained CR+VGPR patients after auto-HSCT.

Diffuse large B-cell lymphoma (DLBCL) is biologically aggressive and most common pathological type of non-Hodgkin's lymphoma (NHL), and about one-third of these patients are refractory or relapsed ultimately. Based on the molecular heterogeneity, DLBCL can be divided into 3 main molecular subtypes: germinal center B-cell like (GCB), activated B-cell like (ABC) and primary mediastinal B-cell lymphoma (PMBL). Arising from B cells at distinct stages of differentiation, these subtypes are also diverse in mechanisms of oncogenic activation. So targeting specific oncogene addictions within the DLBCL subtypes offers a more-precise approach to therapy in comparison with the traditional chemotherapies, undoubtedly bringing a bright hope for the treatment of relapsed/refractory DLBCL. In this article, the advances in diagnosis and therapy of GCB-DLBCL, ABC-DLBCL and PMBL relapsed/refractory diffuse large B-cell lymphoma are summarized.

Objective: To explore the relationship between expression of CD96 and CD123 and prognosis of patients with myelodysplastic syndrome(MDS)., Methods: Eight-nine MDS patients(MDS group) and 20 persons without hematologic disease as controls(Control group) were enrolled. The patients were grouped by the risk. All participants received bone marrow biopsy. Mononuclear cells were extracted, CD34 + CD38 - CD123 + and CD34 + CD38 - CD96 + cells were counted by using flow cytometry. Expressions of 2 type cells in control group, MDS group and its subgroups were analyzed., Results: The proportion of CD34 + cells and CD34 + CD38 - cells in mononuclear cells of patients in MDS group was higher than in control group (P<0.05). The proportions of CD34 + CD38 - CD123 + cells and CD34 + CD38 - CD96 + cells in CD34 + CD38 - cells were significantly higher than that in control group(P<0.05) and the proportion increased with the risk. In the low-and middle-risk group, the rates of complete remission(CR) and partial remission(PR) of patients with CD123 - and CD96 - were higher than those in patients with CD123 + and CD96 + ; in the middle-2 and high risk patients, the PR of patients with CD123 - was higher than that in patients with CD123 + (P<0.05). The CR rate of patients with CD96 - was higher than that of patients with CD96 + (P<0.05)., Conclusion: The differentiation of CD34 + cells in bone marrow of MDS patients is abnormal, and the high expression of CD123 and CD96 cells existes. These findings may partially explain the cause of hematopoietic stem cell malignant clone in MDS patients.

Objective: To observe the efficacy of chemotherapy consisted of bortezomib as main druy in maintenance therapy for recurrence of newly diagnosed MM patients., Methods: The clinical data and outcome of 37 MM patients during 2008-2013 were analyzed retrospectively, the 37 MM patients were divided into 2 group: 19 cases including 13 cases of newly diagnosed MM with symptoms and 6 cases of relapsed refractory MM were enrolled in group A; 17 cases of newly diagnosed MM with symptoms were enrolled in group B. The patients of group A received maintenance therapy consisted of bortezomib plus dexamethasone (VD group), while the patient group B received maintenance therapy consisted of melphalan plus prednisone(MP group), then the therapeutic efficacy of 2 group was compared., Results: The overall response rate(ORR) in VD groupe was 84.2%(16/19), out of which CR rate reached 42%(8/19), PR rate reached 31.6%(6/19), MR rate reached 10.5%(3/19). During median follow-up for 21.8(5-51) months, death occurred, while the ORR in MP group was 52(9/17), out of which CR rate was 23.5%(4/17), PR rate reached 23.5%(4/17), MR rate reached 5.9%(1/17). Druing median follow-up for 16.4(4-39) months, the worteity reaced 64.7%(11/17). The differencr between 2 groups was significant(P<0.05). The median OS time of patients in VD group was 21.6 months, that in MP group was 17.9 months(P<0.05). The median PFS in VD group and MP group were 13.4 and 9.4 months respectively(P<0.001)., Conclusion: The ORR and CR rates of bortezomib maintenance therapy for newly diagnosed and relapsed / refractory MM patients are very high, and its toxicity can be controlled, therefore, the patients need maintenance therapy after remission.

The pathogenesis of diffuse large B-cell lymphoma(DLBCL) has not been fully elucidated, but people realized that the occurrence of DLBCL is caused by the mutual change of genetics and epigenetics with the rapid development of modern biological technology. Epigenetics is to explore the expression changes of genes and proteins without the changes of gene sequences, and this change is reversible and can be inherited. In this review, the latest advances of epigenetic changes in DNA methylation, microRNA regulation, and histone modification for DLBCL are summarized, so as to provide a new strategy to study the early diagnosis, treatment and prognosis of DLBCL.

Splenic marginal zone lymphoma (SMZL) is an uncommon indolent B-cell non-Hodgkin's lymphoma. The clinical features of SMZL are splenomegaly, lymphocytosis and cytopenia in peripheral blood. Immunopheno-typically, the neoplastic cells are typically positive for CD45, CD20, CD79a, PAX5, IgD, BCL-2. The previous common used treatment is splenectomy and chemotherapy. Nowadays, rituximab-based chemotherapy regimens has improved the curative effect dramaticlly. IFN-α with or without ribavirin can be used to treat the patients with HCV infection. Some new target agents such as Bruton tyrosine kinase, NOTCH inhibitor, CAL-101 are under clinical research. This review mainly discuss the progress in the treatment of SMZL.

Background: A survey of early stage follicular lymphoma(FL) revealed that the rigorously staged FL patients at first diagnosis had a better outcome as compared with non-rigorous staged FL patients, but there were no similar reports in China., Objective: To explore the relationship between the rigorous staging at first diagnosis and the prognosis of FL patients at different stages., Methods: The clinical data of 111 patients with newly diagnosed FL from 2008 to 2014 year were collected and analyzed. The rigorous staging included: (1) bone marrow aspiration and biopsy, (2) imaging examination of whole body including CT and ultrasounic scan, or PET/CT, either or both is defined as rigorous staging, or else as non-rigorous staging., Results: The FL patients at I-II stages by rigorous staging showed a superior progression-free survival(PFS) compared with non-rigorous staging patients(P=0.048). For all the patients, the age, serum LDH, bone marrow lesion and more than 3 foci of diameter larger than 3 cm correlated with prognosis in univariate analysis, and multivariate analysis revealed that the age, serum LDH and bone marrow imolvement were the independent prognostic factors., Conclusion: Rigorous staging leads to better outcomes, suggesting that accurate and appropriate testing is important for the patients at the first treatment. The close correlation of bone marrow with prognosis indicates that the evaluation of bone marrow is very important for the daily clinical practice.

Objective: To establish a method for determination of glycosyltransferase and to explore the enzyme A, B glycosyltransferase activity in human serum so as to lay the foundation for the determination of enzyme level and enzyme activity., Methods: The glycosyltransferase activity kit was used to draw phosphate standard curves in our laboratory. The A and B glycosyltransferase activity were determined by the standard curves., Results: The standard curves (y=2671.3x-0.596 R 2 =0.9998) for determing glycosyltransferase activity suitable for use in our laboratory were drawn. At the same time the method was set up for determination of A, B glycosyltransferase in human serum., Conclusion: The establised method of the determination of glycosyltransferase is suitable for common type of enzyme activity and suitable for the A, B glycosyltransferase in human serum.

Multiple myeloma (MM) is a malignancy of terminally differentiated monoclonal B cells, which is characterized by the presentation of malignant plasma cell within the bone marrow and the secretion of monoclonal immunoglobulin. Extramedullary disease (EMD) may be found either at diagnosis or during therapy of these patients. For the patients with EMD, the response to conventional chemotherapy is poor and the prognosis is unfavorable. This review mainly discusses the research progress in the pathogenesis, the clinical feature, the therapy and the prognosis of MM with EMD.

Objective: To investigate the clinical characteristics and prognostic factors of patients with peripheral T cell lymphoma (PTCL)., Methods: The clinical data of 46 elderly PTCL patients admitted in Tianjin Medical University Cancer Hospital from April 2008 to August 2014 were collected, the clinical features, prognostic factors and treatments, as well as followed-up outcome were analyzed retrospectively. Survival analysis was performed by Kaplan-Meier method, and the COX proportional hazard model was used to perform multivariate analysis., Results: The median survival time was 11 months, and the expected 1-year, 2-year and 3-year overall survival rate (OS) was 50%, 36% and 33%, respectively. Univariate analysis showed that the age, ECOG score, Charlson Comorbidity Index Score, the efficacy and course of chemotherapy were all the prognostic indicators affecting the OS and progression free survival (PFS) in this cohort of elderly patients. Multivariate analysis indicated that ECOG score and course of chemotherapy were the independent prognostic indicators affecting the OS and PFS (P < 0.05)., Conclusion: ECOG score and course of chemotherapy are of great significance for predicting the prognosis in elderly PTCL patients. The elderly patients's general condition and completion of a certain intensity of chemotherapy are an important measure to prolong survival time in elderly PTCL patients.

Objective: To analyze the clinical characteristics and prognostic factors of the newly diagnosed Hodgkin's lymphoma (HL) patients., Method: The clinical data of 223 patients with pathologically confirmed and newly diagnosed HL (since January 2001 to December 2012) were collected from Tianjin Medical University Cancer Hospital. The patients' route features, laboratory examination results, clinical manifestations and short-term efficacy were analyzed. Furthermore, the Log-rank test and Cox proportional hazard model were applied to analyze the prognosis. All data were analyzed with SPSS software version 1.0 and graphPad Prism 5., Results: Among the 223 newly diagnosed HL patients, the ratio of male to female was approximately 1.25:1 and the median age was 40 years old (range 3-84 years). The median survival period is 62 months (range 8-189 months). The 5-year overall survival (OS) rate was 92.8% for all 223 HL patients. Multivariate analysis showed that HL originated from mediastinal lymph node region, the extranodal sites were involved, lactate dehydrogenase (LDH) level increased and CR2 was unable to fulfil the CR/CRu which are independent prognostic factors of HL patients (P < 0.05). Therefore, according to the above-mentioned adverse prognostic factors, 223 cases of HL patients were regrouped as low, intermediate and high risk. The HL 5-year OS rate of three groups were 100%, 96.9% and 42.9%, OS in each group was significantly different (P < 0.05)., Conclusion: HL displays a high incidence in young male patients but the curative efficacy is optimal and the survival period is long. The HL originated at mediastinal lymph node region, involved extranodal sites, lactate dehydrogenase (LDH) level increased and CR2 is unable to fulfil the CR/CRu which are independent prognostic factors of HL patients. According to the above-mentioned factors for the initial treatment of patients with HL, risk discrimination can predict the prognosis of patients with HL to a certain extent.

Objective: To investigate the clinical characteristics, treatment and prognosis of solitary myeloid sarcoma (MS)., Methods: The clinical data of 14 solitary MS patients were retrospectively analysed, including their clinical features and treatment, and were evaluated., Results: A total of 14 cases of solitary MS mainly occurred in middle-aged population with the median age 41 years old (17-62 years old). The involved sites were more extensive, including breast, testis, spinal canal, skin, gastrointestinal system, nose and so on. The poorly differentiated cells of small to medium size showed diffuse distribution, relatively consistent morphology and a higher ratio of cytoplasm. The nucleus is in round or oval shape with fine and dense chromatin. Pathological mitosis was easily observed. Expression of MPO, lysozyme, CD43, myeloid-derived cells were positive. Treatment methods included surgery, chemotherapy and stem cell transplantation. Median survival time of 14 patients was 22.5 months; overall survival (OS) was 35.7% (5/14), median disease-free survival reached to 10.4 months on averge (3.5 months to 16 months), and 2-year overall survival (OS) was 50.3%., Conclusion: The incidence of solitary MS is low, with a tendency progressing to leukemia, the chemotherapy regimen of anthracycline+cytarabine combined with radiotherapy can achieve better clinical efficacy.

Objective: To detect the base sequences of all exons and part of introns in the GYPA gene of the glycophorin GPA and to investigate the polymorphism of M, N alleles in Chinese population., Methods: A total of 225 blood sample were randomly colleeted from unrelated Chinese volunteers and were detected by serology techniques. The primers were designed by self, the seguencing of GYPA gene related with sample exon 1-7 full length sequences of bases and intron-1-7 partial sequence was performed, the polymorphism of M, N gene mutation in mucleotide sequence was analysed., Results: The results of M and N genotyping were in agreement with the results of serological detection. The 23rd base of intron-2, the 55th base of intron-3, the 63rd base of intron-4, the 55th, 189th and 190th base of intron-6, the 712th base variation of exon-7 in the gene M and N were used to subdivide the gene M and N into the mutant M103, M201, M202, N101, N102, N103, N104, and N201. At the same time, it was found that 42th and 54th base were mutated, the base T was inserted between 59th and 60th base in the intron-2, the new mutations occurred in the alleles 28, 29, 65 and 102 in intron-3 in this study., Conclusion: The polymorphism of the the Chinese population's GYPA gene occurs in all the exons and partly in the introns. The gene polymorphism of M and N blood group in Chinese population might provide the theoretical basis for the studies of clinical blood transfusion, human population genetics and molecular biology.

Therapy-related myelodysplasia syndromes/acute myeloid leukemia (t-MDS/AML) is a clinical syndrome occurring as a late complication after chemotherapy and (or) radiotherapy, attracting much more attention owing to the improved treatment agents and longer survival of many treated patients. According to the WHO classification of 2008, t-MDS/AML is a serious complication of chemotherapy or radiotherapy given to a malignant or nonmalignant condition consisting of t-AML, t-MDS and t-MDS/myeloproliferative diseases (t-MDS/MPD). This review mainly focuses on the pathogenesis, relationship with primary tumour, treatment and prognosis of t-MDS/AML.

Epstein-Barr virus (EBV) belongs to r-DNA herpes virus family and the first discovered virus associated with human tumors. Numerous studies have found that EBV infection is related to the occurrence of a variety of lymphomas, such as Hodgkin's lymphoma, Burkitt's lymphoma, NK/T-cell lymphoma, HIV-related lymphoma and diffuse large B-cell lymphoma. Recent studies indicated that the gene expression products of EBV latent infections play an important role in the lymphoma, and these findings provide the theoretical basis for the treatment of EBV-related lymphoma. This review mainly discusses the progress in the mechanism and the therapy of EBV-related lymphoma.

The aim of this study was to investigate the effect of alanine solution as α-N-acetylgalactosaminidase enzyme reaction buffer on the enzymatic activity of A antigen. The binding ability of α-N-acetylgalactosaminidase with RBC in different reaction buffer such as alanine solution, glycine solution, normal saline (0.9% NaCl), PBS, PCS was detected by Western blot. The results showed that the efficiency of A to O conversion in alanine solution was similar to that in glycine solution, and Western blot confirmed that most of enzymes blinded with RBC in glycine or alanine solution, but few enzymes blinded with RBC in PBS, PCS or normal saline. The evidences indicated that binding of enzyme with RBC was a key element for A to O blood group conversion, while the binding ability of α-N-acetylgalactosaminidase with RBC in alanine or glycine solution was similar. It is concluded that alanine solution can be used as enzyme reaction buffer in A to O blood group conversion. In this buffer, the α-N-acetylgalactosaminidase is closely blinded with RBC and α-N-acetylgalactosaminidase plays efficient enzymatic activity of A antigen.

The purpose of this study was to evaluate the potential associations between HLA-A, B, DRB1 gene and leukemia. A total of 1186 leukemic patients, including 326 patients with acute lymphoblastic leukemia (ALL), 545 patients with acute myeloid leukemia (AML), 315 patients with chronic myeloid leukemia (CML), and 1234 healthy unrelated donors were typed and were compared in a single centre by using same technique, then the Bonferroni correction method was used to correct the Type I error. The results indicated that as compared with the control,the frequency of HLA-DRB1(*)09 in ALL group significantly decreased (10.87% versus 16.08%; Pc = 0.014, OR = 0.637, 95% CI = 0.487-0.834), while in comparison with control, the frequency of HLA-B(*)18 in CML group was significantly higher (1.28% vs 0.20%; Pc = 0.039, OR = 6.336, 95% CI = 2.066-19.434). The positive and negative relation may exist between certain HLA molecules and leukemia. The negative relation between HLA-DRB1(*)09 and ALL indicated that DRB1*09 might play an important role by a restricted T-cell immune response in the early leukemogenic events, whereas the positive relation between HLA-B(*)18 and CML suggests that the B(*)18 molecules may not actively present leukemia-specific antigens resulting in immune escape. It is concluded that these findings can contribute to developing more appropriate method in leukemic immunotherapy.

CAL-101 is a selective inhibitor of the phosphatidylinositol-3 kinase (PI3K), it inhibits the survival, proliferation and migration of tumor cells by directly inducing apoptosis and inhibiting micro-environmental interactions. It has been determined that the P110δ isoforms of PI3K expressed primarily in cells of hematopoietic lineage, such as B and T cells. This review focuses on the target, mechanism of action, the use and prospect of CAL-101 in tumors of blood and lymph systems.

Lymphoblastic lymphoma (LBL) comprises 2% to 4% of non-Hodgkin lymphomas cases in adults, of which 85% to 90% of LBL in adults is of T-cell phenotype. This study was aimed to evaluate the clinical characteristics and prognostic factors of patients with mediastinal T-LBL. Based on the retrospective analysis of the clinical data of 35 patients with mediastinal T-LBL during the period from January 1998 to January 2011, the clinical characteristics and prognostic factors of mediastinal T-LBL were summarized. The results showed that the total of 35 patients were identified (male 24 and female 11), with a median age of 19 (5 - 52) years. The majority of patients were in stage III/IV, 16 cases (45.7%) presented bulky mediastinal mass. Intrathoracic effusions (pleural, pericardial) were not uncommon (62.9%). Overall survival rate (OS) and progression-free survival rate (PFS) at 3 years for the entire cohort were 36% and 24%, respectively. OS and PFS at 5 years were 25% and 16.7%, respectively. Anemia at diagnosis were an important, independent predictor of OS (P = 0.048). Bulky mass (P = 0.048), superior vena cava syndrome (P = 0.021), and abnormal PLT count at diagnosis was the independent prognostic factors for PFS (P = 0.021). It is concluded that the patients with primary mediastinal T-LBL are characterized by a low incidence, bad prognosis, and short survival. For patients accompanying with anemia, bulky mass and superior vena cava syndrome, their prognosis is worse.

This study was aimed to investigate the expression levels of CXCR4 and VEGF in serum of patients with DLBCL and their clinical significances. The peripheral blood of 44 patients with newly diagnosed DLBCL and 20 healthy adults as a control group were chosen for study. And the expression levels of CXCR4 and VEGF in serum were detected by ELISA. The results showed that the expressions of VEGF and CXCR4 in DLBCL patients were higher than those in the control group (P < 0.05). The expression of VEGF was positively correlated with the expression of CXCR4 in DLBCL patients, and the correlation coefficient was 0.743 (P < 0.05). The VEGF expression in DLBCL patients was correlated with LDH, immunotyping, the number of extranodal involvements, Ann Arbor staging and ECOG performance score; while the expression of CXCR4 was correlated with LDH, immunotyping, the number of extranodal involvements and Ann Arbor staging. Univariate analysis showed that LDH, extranodal involvements, immunotyping, Ann Arbor staging, CXCR4 and VEGF were associated with OS. Multivariate analysis showed that the immunotyping and CXCR4 expression independently associated with OS. It is concluded that both expression levels of VEGF and CXCR4 are significant higher than those in the control group. CXCR4 expression positively correlates with VEGF expression and displays a prognostic significance for OS. This study suggests that combined targeting VEGF and CXCR4 may become a novel therapeutic strategy for diffuse large B cell lymphoma.

Aim of this study was to explore the effects of cryopreservation on biological characteristics of wharton's jelly derived mesenchymal stem cells (WJ-MSC), and to provide experimental evidence for clinical applications and the establishment of WJ-MSC bank. Primary WJ-MSC were produced by umbilical cord tissue culture in vitro. Fifth passage of WJ-MSC acquired by continuous cell culture were mixed with cryoprotectants, frozen in -80°C refrigerator and stored in liquid nitrogen. After the cryopreserved WJ-MSC were thawed, the first passage of WJ-MSC was obtained through cell culture and was taken as the 1st preserved passage (PP1). Thus, PP2-PP15 WJ-MSC were obtained by continuous cell subculture. The 1st control passage (CP1) to 15th passage (CP15) represented the 6th passage to 21st passage WJ-MSC acquired by subculturing in non-cryopreserved group. The biological characteristics of WJ-MSC from cryopreserved and control group, including the recovery rate of nucleated cells, trypan blue exclusion, CCK-8 activity, cell apoptosis, cell adherence, proliferation index, cell surface antigen, cell cycle and the capacities of induced differentiation into adipocyte, osteoblast and neuron, were detected and compared. The results indicated that the recovery rate of nucleated cells of cryopreserved WJ-MSC was 98.2%, trypan blue exclusion rate was 94.3%, CCK-8 activity was 91.4%, apoptotic rate was 3.9%, and the adherence rate was 92.6%. There was a statistically significant difference in proliferation index between PP1 and CP1 (P < 0.05), but there were no statistically significant differences between PP2-PP15 and their corresponding controls. The subculture cells highly expressed CD29, CD44, CD71, CD73, CD90, CD105, CD166 and HLA-ABC, and lowly expressed CD34, CD45 and HLA-DR. The expressions of above-mentioned surface antigens were not different statistically between two groups. The proliferation latency and logarithm proliferation of the subculture cells between two groups were also not different. After induced differentiation into adipocyte, osteoblast and neuron, the staining with oil red O, alkaline phosphatase and neuron-specific enolase was performed respectively, and the positive degrees were not clearly different macroscopically between two groups. Relatively high levels of triglyceride, alkaline phosphatase, and neuron-specific enolase in relevant cells could be detected, but had no significant differences between two groups. It is concluded that some WJ-MSC (< 10%) are damaged after cryopreservation, and the biological characteristics of WJ-MSC in cryopreservation group keep constant, as compared with that in non-cryopreservation group.

Alteration in the balance between cell apoptosis and proliferation is one of the pathophysiological mechanisms of the myelodysplastic syndromes (MDS). The question of whether the excessive apoptosis and/or proliferation predominantly involve the subset of progenitor cells (CD34(+) cells) or mature cells (CD34(-) cells) remains a controversial issue. This study was purpose to analyze the apoptosis and proliferation status of CD34(+) and CD34(-) cells in bone marrow (BM) of patients with MDS, to investigate the pathogenesis of MDS and to determine the relation of apoptosis and proliferation status of CD34(+) and CD34(-) cells with prognosis of MDS. The proportion of CD34(+) cells, the apoptosis and proliferation ratio (A/P) of CD34(+) and CD34(-) cells in BM of 40 patients with MDS, including 20 cases of high-risk MDS and 20 cases of low-risk MDS, and 10 normal persons as control were detected by flow cytometry; the influence of CD34(+) and CD34(-) cell apoptosis and proliferation levels on prognosis of MDS was evaluated by univariate and multivariate analysis of survival. The results showed that the proportion of CD34(+) cells in BM of high-risk MDS patients was significantly higher than that in BM of low-risk MDS patients and in normal BM [(1.92 ± 0.10)%, (1.09 ± 0.04)%, (1.03 ± 0.05)% respectively]. The apoptotic rates (AR) of both CD34(+) and CD34(-) cells were significantly higher in low-risk MDS [(54.75 ± 2.18)%, (80.36 ± 1.68)%] than in high-risk MDS [(24.87 ± 2.69)%, (23.12 ± 1.23)%] and in normal BM [(18.51 ± 2.74)%, (20.98 ± 2.21)%]. When compared between CD34(+) cells and CD34(-) cells in low-risk MDS, a greater AR of CD34(-) cells was found. However, the higher proliferative rate of CD34(+) cells was observed in high-risk MDS. In low-risk MDS, a higher A/P ratio was found in CD34(-) cells than in CD34(+) cells; whereas this ratio was equalized or inverted in high-risk MDS. In addition, the survival and prognosis correlated significantly with AR of CD34(+) cells. It is concluded that the early MDS is predominantly associated with excessive apoptosis of the mature CD34(-) cells. The proliferation rate of cells increases with the disease progression in MDS subsets, especially, in the subset of CD34(+) cells. Surprisingly, the apoptosis of CD34(+) cells may be a useful prognostic factor, and the inhibition of apoptotic mechanisms may induce the transformation of MDS to leukemia.

NK/T cell lymphoma, a rare type of non-Hodgkin's lymphoma, is a highly aggressive disease with poor prognosis. Clinically, it is classified into nasal, non-nasal, and aggressive lymphoma/leukemia subtypes. They are characterized by geographic distribution and are universally associated with Epstein-Barr virus (EBV) infection. Due to its low occurrence and dismal clinical outcome, no therapeutic strategy is currently identified in this disease. Combined chemotherapy and radiotherapy have better effects for stage I/II nasal NK cell lymphoma. As for stage III/IV nasal NK cell lymphoma and non-nasal, and aggressive subtypes, chemotherapy is the main treatment method. Recently, some studies have demonstrated promising outcomes in the selected cases by high-dose chemotherapy supplemented with auto- or allo-HSCT.

The objective of this study was to evaluate the value of morphologic diagnosis for acute leukemia (AL), to explore the relation of morphologic diagnosis with immunology, cytogenetics and molecular biology diagnosis of AL and to analyze the onset characteristics of AL in 10 years. The samples of bone marrow and peripheral blood from 233 newly diagnosed cases of AL were collected during 2001-2011 years; the morphologic examination and immunologic, cytogenetic and molecular biologic examination (ICM) were carried out, the consistency of morphologic diagnosis with ICM diagnosis was compared, the onset characteristics of AL was analyzed. The results showed that: (1) the consistent rate of immunology, cytogenetics, molecular biology diagnosis with morphologic diagnosis was 84.3%. The order of consistent rat was AUL, M0 < M1 < HAL < M4 < M2 < M3 < M5 < ALL < M6, M7, AP; (2) Misdiagnosis always occurred among AUL, M0, M1, ALL and HAL or among M2a, M3v, M4 and M5. (3) In 233 cases, the highest ratio of blast was observed in M1 (92.5%), while the lowest ratio of blast was observed in M2 (49.5%). (4) AL occurred more frequently in males than that in female (147:86). (5) AL occurred in patients aged from 1 to 88 years. The median age was 41.5 for AUL, 40.8 for M0, 43.4 for M1, 46.3 for M2, 33.8 for M3, 42.6 for M4, 48.8 for M5, 77.3 for M6, 2.5 for M7, 65.0 for AP, 29.1 for ALL and 40.3 for HAL. (6) The number of patients in the later five years (139 cases) was significantly greater than that in the first five years (94 cases), especially the patients with M1, M2, M3, M4, and M5. It is concluded that morphologic diagnosis has important clinical value in the MICM diagnosis of AL. Attaching importance to the confusing cell morphology and onset characteristics of AL can improve the diagnostic accuracy.

This study was aimed to evaluate whether human mesenchymal stem cell (MSC) and the Stro-1 positive subgroup have inducing immune tolerance effect in mouse skin graft model. Human MSC were isolated and cultured from bone marrow-derived mononuclear cells of healthy adults, and Stro-1 positive cells were sorted out. Female C57BL/6 mice and female BALB/c mice were respectively used as donors and recipients in skin allogenic graft model. The recipients were divided randomly into 4 groups: (1) Stro-1(+) MSC group: 2×10(6) Stro-1(+) MSC were injected into the irradiated recipient mice before skin graft. (2) MSC group: 2×10(6) MSC were injected into the irradiated recipient mice before skin graft. (3) Irradiation control group: the recipient mice were just irradiated before skin allogenic graft. (4) Congenic control group: the irradiated BALB/c mice received the skin from the congenic mice. The survival time and pathologic changes of skin grafts were observed by macro- and microscopy with HE staining. The transforming growth factor β1 (TGF-β1) concentration in plasma of recipient mice was measured by ELISA before and after grafting. The results indicated that the survival time of skin grafts in the MSC group was (12.13 ± 3.34) d, which was not notably longer than the irradiation control group (11.38 ± 1.01) d. The survival time of skin grafts was significantly prolonged in the Stro-1(+) MSC group (30.68 ± 5.89) d, as compared with the irradiation control group and the MSC group, respectively; the pathologic examination of skin grafts showed a clear structure. After grafting, the TGF-β1 concentration in plasma of recipient mice was almost the same as before grafting in the irradiation control group and the congenic control group, but it significantly increased in the MSC group and the Stro-1(+) MSC group. It is concluded that the Stro-1(+) MSC induce greater immune tolerance than the unsorted MSC, and significantly prolong the survival time of skin grafts in vivo, while TGF-β1 does not contribute to the immune tolerance.

This study was purposed to detect the expression of transforming growth factor β1 (TGF-β1) and its receptors (TGF-βR) and to investigate their roles in pathogenesis of immune thrombocytopenic purpura (ITP). The expressions of TGF-β1 and their receptors TGF-βRI, TGF-βRII and TGF-βRIII in the peripheral blood of patients with ITP and healthy persons were detected by the real-time PCR, and differences of their expression levels were analysed. The results showed that the expression of TGF-β1 and TGF-βRII mRNA in ITP patients was significantly higher than that in the healthy controls, while the TGF-βRI mRNA expression in ITP patient was significantly lower than that in the controls. The expression of TGF-βRIII was not statistically different between the two groups. It is concluded that TGF-β1 and its receptors including TGF-βRI and TGF-βRII express abnormally in the peripheral blood of ITP patients, which suggests that the TGF-β signaling pathway probably play a vital role in the pathogenesis of the ITP.

The effects of conventional treatment for myelodysplastic syndrome (MDS) are not remarkable to date, while only a minority of patients was eligible for allogeneic stem cell transplantation. As epigenetics plays a significant role during the occurrence and development of MDS, and histone deacetylase inhibitors (HDACI), a class of gene expression modulating drugs, are currently being developed for therapy of several types of solid tumor, more attention is paying to HDACI as potential therapy of MDS. This review summarizes briefly the rationale for HDACI use in MDS, the common mechanism of HDACI, the present state of the clinical efficiency, and future development in this field.

Lenalidomide is an immunomodulatory, antiangiogenic drug that is a structural analog of thalidomide. Studies showed that lenalidomide may work through various mechanisms in hematologic malignancies. These mechanisms involved direct cytotoxicity as well as through indirect effects on tumor immunity etc. It is approved by the Food and Drug Administration for treatment of multiple myeloma and myelodysplastic syndromes, and proved to have a good efficacy. Recent studies demonstrate that oral lenalidomide alone produces durable responses with manageable adverse events in patients with non-Hodgkin's lymphoma, relapsed/refractory Hodgkin's lymphoma, chronic lymphocytic leukemia and older patients with acute myeloid leukemia etc, warranting further investigation of treatment for these patients. This review focuses the related studies and the latest progression about lenalidomide in hematological malignancies in order to provide some references and help to the use of lenalidomide for the treatment of hematological malignancies.

The aim of this study was to explore the effect of mesenchymal stem cell (MSC) conditioned medium (MSC-CM) on proliferation, migration and adhesion of human umbilical vein endothelial cell (CRL1730) and its mechanism. Isolation and purification of MSC were performed with the classic adhering method, the surface markers (CD29, CD90, CD45 and CD34) in MSC were detected by flow cytometry. MSC were treated and cultured for 3 d, the MSC-CM or MSC overexpressing stem cell-derived factor-1 (SDF-1) conditioned medium (Ad-SDF-1-MSC-CM) were collected. Subsequently, CRL1730 cells were treated respectively with 2% FBS-DMEM, 15% FBS-DMEM (control group), MSC-CM or Ad-SDF-1-MSC-CM for 24 h, the proliferation of CRL1730 cells was detected by MTT method. CRL1730 cell migration in vitro was performed by using wound healing system. The adhesion ability of CRL1730 cells was analyzed by microscope. The results indicated that the CRL1730 cells treated with Ad-SDF-1-MSC-CM showed greater proliferative capacity than CRL1730 cells treated with MSC-CM. While adding with AMD3100 5 µmol/L, the blocker of CXCR4, the CRL1730 proliferation mediated by Ad-SDF-1-MSC-CM was significantly reduced. Meanwhile, compared with MSC-CM, Ad-SDF-1-MSC-CM had greater effects for promoting CRL1730 migration and enhancing adhesion ability of CRL1730 cells, these effects were significantly inhibited by AMD3100. It is concluded that MSC-CM promotes the migration and adhesion ability of CRL1730 cells through SDF-1 expressed by MSC.

The objective of this study was to explore the effect of mesenchymal stem cells (MSC) on HL-60 proliferation and its molecular mechanism. HL-60 cells co-cultivated with MSC were used as experiment group, and the cells cultivated solely were taken as control group. HL-60 cells in the two groups were counted at different time. The time-quantity curve was drawn. The cell cycle and apoptosis ratio of HL-60 cells were compared between the two groups and expressions of Survivin and Bcl-2 protein were detected by flow cytometry. The results showed that HL-60 cells cultivated with MSC were obviously inhibited, especially on day 5 and 7 (P < 0.01). HL-60 cells were distributed on the phase of G(0)/G(1) [control group (47.0 ± 9.0)% vs experiment group (70.0 ± 16.0)%, P = 0.003], and apoptotic peak appeared. Both of Survivin and Bcl-2 protein expressions in HL-60 cells decreased [Bcl-2 protein in control group (63.0 ± 9.1)% vs experiment group (50.0 ± 14.1)%, P = 0.045; Survivin in control group (94.0 ± 9.3)% vs experiment group (77.0 ± 11.8)%, P = 0.006]. It is concluded that the MSC can inhibit HL-60 cell proliferation, and promote HL-60 cell apoptosis.

Hematopoietic stem cells (HSC), the well-characterized adult stem cells both in the markers and function, show tremendous therapeutic potential. However, the level of hematopoietic stem cells in bone marrow is very low, which makes it difficult to work with. Based on cell surface markers and dye staining, HSC can be isolated from bone marrow and peripheral blood by using magnetic-activated cell separation and fluorescence-activated cell sorting. Over 40 years of research, many surface markers have been identified to purify mouse HSC, and CD34(-)LSK cells are regarded as the mostly used enrichment of HSC in mouse bone marrow. The purpose of this review is to provide insight into the advance of these markers, and isolation and purification of HSC.

The study was aimed to investigate the expression of stromal cell derived factor (SDF-1) in bone marrow (BM) and its relation with apoptosis of BM CD34(+) cell and angiogenesis in myelodysplastic syndrome (MDS). 40 patients with MDS were divided into low-risk group and high-risk group according to IPSS score system. BM samples were collected. SDF-1 levels, the apoptosis of CD34(+) cells and microvessel density (MVD) of BM were detected by ELISA, flow cytometry and immunochemistry, respectively. The results showed that the SDF-1 level in MDS patients was significantly higher than that in normal controls(p < 0.05), and SDF-1 level in low-risk group was significantly higher than that in high-risk group. Apoptosis of CD34(+) cells significantly increased in low-risk group compared with other groups (p < 0.05). MVD in BM biopsy significantly increased in high-risk MDS group (p < 0.05), compared with low-risk MDS group which also had higher MVD than the control group (p < 0.05). Positive correlation was found between apoptosis of CD34(+) cells and SDF-1 levels in low-risk group, and SDF-1 level and MVD in high-risk group. It is concluded that the expression of SDF-1, apoptosis of BM CD34(+) cells and MVD were significantly abnormal in MDS patients, especially in different risk group, suggesting that SDF-1 level is related to cell apoptosis and angiogenesis.

Many studies indicate that lymphoid neoplasms are related with chromosome translocations and the molecular alterations involving in the cell cycle and/or apoptotic pathways. However, survival of B and T tumor cells also depends on interactions of these cells with the accompanying cells comprising the lymphoma microenvironment. Immune cells, stromal cells and numerous molecular together make up the microenvironment and have functional interaction with tumor cells, promoting tumor growth and drug resistance. Different types of lymphoma have various clinical courses, therapy responses and prognoses, which show a close relationship with the microenvironment. This review summarizes several components of lymphoma microenvironment including macrophages, adhesion molecules and chemokines and the roles of microenvironment in classic non-Hodgkin's lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, suggesting that the microenvironment influence the prognosis of lymphoma, targeting microenvironment may be a potential method in lymphoma therapy.

In order to explore the effect of VEGF on mesenchymal stem cell (MSC) proliferation and its possible signal transduction mechanism, MSC culture was performed with the classical bone marrow adhering method; characteristics of passage 3 rat MSC (P3MSC) was identified through multi-differentiation and surface marker assay (CD34, CD45, CD90, CD29); P3MSC were treated with 20 ng/ml VEGF, and the effect of VEGF on the MSC proliferation was measured during 12, 36 and 72 hours by MTT assay. Subsequently, P3MSC were treated with extracellular-signal regulated kinase (ERK1/2) inhibitor PD98059 (50 µmol/L) or p38 mitogen-activated protein kinase (p38MAPK) inhibitor SB203580 (30 µmol/L) for 30 minutes, the culture medium was replaced with new medium including 20 ng/ml VEGF. After 72 hours, the effect of PD98059 or SB203580 on MSC proliferation mediated by VEGF was measured by MTT assay. The result showed that the cultured MSC expressed PDGFR-α, PDGFR-β and NRP1, but did not express VEGF-R (Flk1 and Flt1). The MSC had the multi-differentiation ability and displayed the characteristics of CD90+ (96.7%), CD29+ (94.6%), CD34- (0.79%) and CD45- (0.84%). The MSC proliferation rate increased gradually with prolonging of the functioning time of 20 ng/ml VEGF, and MSC proliferation rate may reach to maximum value after treating with 20 ng/ml VEGF for 72 hours. The effect of VEGF on MSC proliferation was found to be abolished, even was under level of control group after treating with PD98059 or SB203580 for 30 minutes. Furthermore, the inhibitory effect of PD98059 on MSC proliferation was obviously higher than that of SB203580. It is concluded that the VEGF can promote MSC proliferation, and its possible mechanism may relate to ERK1/2 pathway.

The aim of this study was to investigate the best method to preserve human bone marrow cells and the effectiveness of long term cryopreservation at -80 degrees C. The human bone marrow cells in 20 samples were firstly frozen by a programmed freezer or -80 degrees C refrigerator, and then were preserved in liquid nitrogen with DMSO-AuP (10% dimethylsulfonamide, 10% autologous plasma) or DMSO-HES-HuA (5% dimethylsulfonamide, 6% hydroxyethyl starch, 4% human serum albumin) as cryoprotectant for 21 to 25 years. They were thawed in 38 degrees C. The cell sample frozen in -80 degrees C refrigerator was frozen at a low frozen speed of 1 degrees C/min which was the same as the programmed freezer before -30 degrees C. Before detection the bone marrow cells were taken from liquid nitrogen and were thawed in 38 degrees C, then the suspension of bone marrow cells was prepared for detection. The cell morphology and recovery rate of erythrocytes, nucleocytes and platelets; the recovery rate of hematopoietic stem progenitors cells, as well as mesenchymal stem cells were determined. The results showed that the protective effectiveness of DMSO-HES-HuA was better than DMSO-AuP. The mature erythrocytes were destroyed lightly [(3.5 +/- 1.5)% versus (12.6 +/- 4.8)%], the hemolysis rate was lower [(3.3 +/- 1.6)% versus (23.1 +/- 5.1)%]. Osmotic fragility of erythrocytes in the former was not changed, but was dropped in the latter. The recovery rates of red cell, platelet, granulocyte-macrophage colony forming units and long term culture-initiating cells were higher in the former than that in the latter [(96.1 +/- 1.8)%, (70.0 +/- 9.5)%, (49.2 +/- 10.9)%, (54.2 +/- 13.8)% versus (76.3 +/- 5.6)%, (52.7 +/- 8.1)%, (43.5 +/- 12.3)%, (47.2 +/- 13.6)% respectively]. With each kind of cryoprotectant or frozen method, the frozen MSC could keep the original growth properties. With the same cryoprotectant and different frozen method, the cryopreservative effectiveness was not different. The influence of the cryoprotectant prescriptions and the frozen methods on the cryopreservative effectiveness was little. It is concluded that the human bone marrow cells with DMSO-AuP or DMSO-HES-HuA as cryoprotectant, frozen by a programmed freezer or -80 degrees C refrigerator, could be then preserved in liquid nitrogen for long time. When the preserving time was as long as 21 to 25 years, the morphology, the recovery rate and the activity of various kinds of cells were still good. The method of freezing by -80 degrees C refrigerator with 5% DMSO-6% HES-4% HuA and preserving in liquid nitrogen would be convenient, cheap and easily-manipulated for preservation of the human bone marrow cells.

This study was aimed to compare the influences of bone marrow mesenchymal stem cells (BMMSCs) from patients with acute myeloid leukemia (AML), AML patients with complete remission (CR) and non-leukemia patients on HL-60 cells. The HL-60 cells were divided into three groups: group of co-cultivation with BMMSCs of AML patients, group of co-cultivation with BMMSCs of AML patients with CR and group of co-cultivation with BMMSCs of non-leukemia patients. The count of HL-60 cells, the CD11b and survivin expression of HL-60 cells, the cell cycle distribution of the HL-60 cells in 3 groups were compared by flow cytometry, the morphology and differentiation rate of HL-60 cells in 3 groups were observed and compared by microscopy. The results showed that there were no differences in HL-60 cell count at five and seven days, in HL-60 distribution at the G(0)/G(1) phase, in survivin and CD 11b expressions in 3 groups. All cells of 3 groups began to mature, and the differentiation rates in 3 groups were 18.0 +/- 3, 17.0 +/- 1.3 and 19.0 +/- 2.0 respectively, therefore there were no significant differences between the 3 groups (p = 0.23). It is concluded that there is no influence of BMMSCs in 3 groups on the proliferation and differentiation of HL-60 cells.

This study was aimed to compare partial biological characteristics of bone marrow mesenchymal stem cells (BMMSCs) in AML patients, AML patients with complete remission (CR) and non-leukemia patients. The bone marrow (BM) MSCs were divided into 3 groups: group of MSCs from AML patients, group of MSCs from AML patients with CR, group of MSCs from non-leukemia patients. The morphologic features of MSCs were observed by light microscopy; CFU-F numbers of MSCs were counted after Wright-Giemsa staining in situ; the fusion times of MSCs were determined; the growth curves of MSCs were drawn by counting cell numbers; the immunophenotypes and cell cycle of MSCs were detected by flow cytometry; the DI values of MSCs were calculated. The results showed that the morphologic features of MSCs in 3 groups did not display difference; there was significant difference (p < 0.01) of CFU-F numbers in 3 groups, while CFU-F number of MSCs in AML group was minimal; there was significant difference of MSC fusion time in 3 groups, while fusion time of MSCs in AML group was most long; the growth curves of MSCs in 3 groups were similar; MSCs in 3 groups highly expressed CD105 and CD106, but not expressed CD45; the cell distribution ratios at phase of G(0)/G(1) for MSCs in 3 groups were 89.9 +/- 4.0%, 90.2 +/- 3.0% and 91.0 +/- 3.0% respectively; the DI values of MSCs in 3 groups were between 0.9 and 1.1. It is concluded that no significant difference of biological characteristics of the second generations of MSCs is found between those in leukemia and non-leukemia patients.

The aim of this study was to explore the difference of MSC migration mediated by SDF-1/CXCR4 axis through Boyden chamber in vitro migration assay. The SDF-1 density-dependence of MSC migration was observed. Subsequently, the effects of different blocking agents on hSDF-MSC migration were observed after MSC were treated with 50 nmol/L wortmannin, 10 micromol/L LY294002, 50 micromol/L PD98059, 10 micromol/L U73122, 126 micromol/L AMD3100 and 50 nmol/L verapamil respectively. The results showed the efficiency of MSC migration increased gradually with the increasing of hSDF-1 density. And after MSCs treatment with 50 nmol/L wortmannin, 10 micromol/L LY294002, 50 micromol/L PD98059, 10 micromol/L U73122 and 126 micromol/L AMD3100 respectively, the ability of MSC migration decreased. The ability of MSCs migration obviously decreased when MSCs were treated with U73122, AMD3100. It is concluded that the SDF-1/CXCR4-mediated MSC migration may be related to mitogen-activated protein kinase (MAPK), phosphatidylinositol phospholipase C (PI-PLC) and protein kinase (PKC) signal pathways.

This study was aimed to investigate whether the thrombopoietin (rhTPO) may facilitate myelofibrosis or not. The modified Dexter culture system with various concentrations of rhTPO was used to culture the stromal cells in vitro; the proliferative activity of cells was detected by MTT method; the morphologic changes were observed by light and scanning electron microscopy; the staining changes of ALP, PAS, AS-D NCE and IV type collagen were observed by cytochemistry method; the changes of fibronectin, laminin and IV type collagen were assayed by immunohistochemistry method; the cell surface antigens were assayed by flow cytometry. The results indicated that rhTPO could promote the proliferation of stromal cells which was related to the concentrations of rhTPO. Proliferative activity of stromal cells increased with increasing of rhTPO concentration, and was not related to the exposure time. On day 3 stromal cells adhered to the wall, and became oval. On day 7 stromal cells turned to fusiform and scattered dispersively. On day 12 to 14 these cells ranged cyclically and became long fusiform. Cells covered 70%-80% area of bottle bottom at that time. By day 16 to 18 these cells covered more than 90% area of bottom and ranged cyclically. They displayed the same shape as fibroblasts. By light microscopy with Wrights-Giemsa staining, fibroblasts predominated morphologically, few macrophages, endothelial cells and adipose cells were found. There were no significant differences between experimental group and control group. On day 14 to 42 the adherent cells were positive with PAS staining, poorly positive with ALP and naphthol AS-D chloroacetate esterase (AS-D NCE) staining, and the difference in cytochemistry was not significant between two groups. When these cells were dyed with Masson's trichrome and Gomori's staining, neither collagen fibers nor reticular fibers were positive, but fibronectin, laminin, and collagen type IV appeared positive stronger in experimental group than those in control. The expressions of these molecules were not dependent on culture time. By scanning electron microscopy microvilli and fibers on cell surface appeared more and more, monolayer cells evolved into multilayer cells, and newly-formed fibroblasts appeared gradually as culture time prolonged. These alterations were not different among various groups. The expressions of CD34, CD45, CD105, CD106, and CD166 were not affected obviously by rhTPO. It is concluded that rhTPO had no effects on histochemical properties of stromal cells. Fiber staining and scanning electron microscopic examinations revealed that rhTPO can not facilitate fiber formation of stromal cells. But rhTPO may be able to augment the expressions of fibronectin, laminin and collagen type IV of stromal cells. Therefore it is still necessary to follow up the patients for a long time, who have received rhTPO therapy clinically.

This study was aimed to establish the quantitative analysis of hIL-2 in culture supernatant by multifunctional Luminex 100. The lymphocytes were separated from ACD-anticoagulated peripheral blood by density gradient method. The lymphocytes were stimulated with PHA for 48 hours, and frozen at -20 degrees C The relative fluorescence units of standard preparations and samples were detected by multifunctional Luminex 100, and the sample concentrations were calculated by standard curve. The results indicated that the regression equation of standard preparation is Lg (RFU) = 1.547 + 0.867 LgC. ANOVA F = 301.7427, p < 0.05 (nu = 6). The analysis of variance showed F = 301.7427, p < 0.05 (nu = 6). The test of regression coefficient showed t = 17.3707 (nu = 6), p < 0.05. It is concluded that method for induction and measurement of human IL-2 in vitro is established. The standard curve established by this way is statistically significant. There is linear relationship between the concentration of hIL-2 and fluorescence intensity.

The myelodysplastic syndromes (MDS) include a diverse groups of clonal and potentially malignant bone marrow disorders. Evidences exist that microenvironment cells from MDS marrow show functional abnormalities, which may be relevant to the incidence of such a disease. Mesenchymal stem cells (MSCs) are a very important component of hematopoietic microenvironment. This study was supposed to investigate the biological characteristics and functions of MSC derived from patients with MDS in low-risk. MSCs from bone marrow samples of 11 low-risk MDS patients were isolated, cultured and expanded. Morphology, immunophenotype and osteoblasts differentiation were analyzed. Their capacity of proliferation and hematopoietic supporting in vitro were measured. A real-time quantitative reverse transcriptase polymerase chain reaction method (RQ RT-PCR) was used for detecting the expression levels of relative cytokines and chemokines in MSC. MSCs from healthy donors were used as controls. The results showed that the culture-expanded cells from MDS patients displayed a typical fibroblast-like morphology. Cells were positive for SH2 (CD105), SH3 (CD73), Thy-1 (CD90), while negative for CD34 and CD45. After induction, these cells could differentiate into osteoblasts. The proliferative ability of MSCs in MDS patients were not different from those of MSC isolated from normal bone marrow (p > 0.05), however, their capacity of hematopoietic supporting in vitro were significantly weaker (p < 0.05). RQ RT-PCR detection indicated that the SDF-1 gene expression level in MSCs of low-risk MDS patients was significantly higher than that in MSC derived from healthy donors (p < 0.01). It is concluded that the abnormal function of MSC influences the regulation of hemotopoiesis in the bone marrow microenvironment of MDS patients. It is worthy to further investigate the new clue in etiological mechanism and therapeutic strategies for MDS.