Analysis of Clinical Pathological Features and Prognosis in Young Patients with Diffuse Large B Cell Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To explore the clinical pathological features of the patients with diffuse large B cell lymphoma (DLBCL) and their prognostic factors.@*METHODS@#The prognosis of the clinical pathological features and their influence on prognosis of 177 patients diagnosed as DLBCL at the first visit from January 2013 to May 2017 in our hospital were analyzed retrospectively.@*RESULTS@#The univariate analysis showed that overall survival (OS) and progression-free survival (PFS) were associated with later Ann Arbor stage (Ⅲ-Ⅳ) ( P＜0.01, P＜0.05), high performance status (ECOG score 2-4) (P＜0.01, P＜0.05), extranodal involvement ＞1 (P＜0.01, P＜0.05), elevated LDH level (P＜0.01, P＜0.05). B symptom (P＜0.05) and elevated β2-MG level (P＜0.05) also influenced OS. COX multivariate analysis showed that the elevated β2-MG level (P＜0.05) and later stage (Ⅲ-Ⅳ) (P＜0.05) have an independent influence on OS, later stage (Ⅲ-Ⅳ) (P＜0.05) also independently influenced PFS. The patients with high aaIPI score (2-3) and bone marrow involvement before treatment had poor OS (P＜0.01, P＜0.01) and PFS (P＜0.05, P＜0.01).@*CONCLUSION@#Elevated β2-MG level can independently influence OS, and later stage (Ⅲ-Ⅳ) can independently influence both OS and PFS. High aaIPI score (2-3) and bone marrow involvement before treatment have an inferior influence on OS and PFS.

Establishment of A20 Murine B Lymphoma Transplantation Model Labeled with Luciferase / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To establish the animal model of luciferase-transfected A20 murine B cell lymphoma, so as to provide experimental tools to explore the effect of graft versus tumor.</p><p><b>METHODS</b>Luciferase- labeled A20 cells were cloned with puromycin selection. Transfected A20 cells and CBL/6 bone marrow were inoculated into the irradiated BALB/c mice by injection in tailvein to establish the transplantation model. The bioluminescent imaging technique was used to monitor the tumor growth, and then the survival, body weight, tumor formation and pathological characteristics of target organs were observed.</p><p><b>RESULTS</b>A20 cell line stably expressing luciferase gene was successfully obtained. The the bioluminicent imaging found that the tumor luminescence could be observed on day 8 of A20 cell inoculation, and the mean fluorescent intensity was increased along with the tumor growth. Compared with the BMT group, the survival rate and body weight of BMT+A20-Lucmice were decreased significantly. General anatomy showed the tumor mainly formed in the liver and spleen.</p><p><b>CONCLUSION</b>A mouse transplantation model with luciferase- transfected A20 cells has been successfully established, thus laying a foundation for investigation of graft-versus-tumor.</p>

Effect of G-CSF in vitro Stimulation on Distribution of Peripheral Lymphocyte Subsets in the Healthy Persons / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of granulocyte-colony stimulating factor (G-CSF) in vitro stimulation on the distribution of lymphocyte subset in healthy human.</p><p><b>METHODS</b>Peripheral blood mononuclear cells (PBMNCs) were collected from 8 healthy volunteers by density gradient centrifugation on Ficoll-Paque. In vitro 200 ng/ml G-CSF or 200 ng/ml G-CSF plus 10 µg/ml ConA directly act on PBMNCs, then the colleted cells were cultivated for 3 days. Lymphocyte subsets were stained with the corresponding fluoresce labeled antibodies and detected by flow cytometry.</p><p><b>RESULTS</b>The levels of T cells in G-CSF group and G-CSF+ConA group were both higher than that in the control group (P<0.001, P<0.05). However, there were not significantly different in B cells and NK cells levels among the 3 groups. Furthermore, analysis of the effect of G-CSF on T cell subsets indicated that the levels of CD4T cells and CD8T cells in G-CSF group were both significantly higher than those in control group (P<0.01, P<0.05), Treg cells was not different between G-CSF and control group. Compared with the control group, the level of CD4T cells, CD8T cells and Treg cells in G-CSF+ConA group significantly increased (P<0.05, P<0.01, P<0.01). Analysis of G-CSF receptor (G-CSFR) expression showed that G-CSFR expression on T cells in G-CSF+ConA group dramatically increased, as compared with control group (P<0.01).</p><p><b>CONCLUSION</b>The levels of CD4T cells and CD8T cells in healthy human peripheral blood can be increased by G-CSF stimulation. ConA can enhance the level of T cells and induce G-CSFR expression on T cells.</p>

Ascending input from dorsal nuclei of lateral lemniscus mediates the plasticity of inferior colliculus neurons in mice / 生理学报

The auditory system has the ability to adjust its structure and function as the environment changes, which is called auditory plasticity. In the auditory system, inferior colliculus (IC) is an important relay station, which accepts the ascending inputs from dorsal nuclei of lateral lemniscus (DNLL). The present study was aimed to investigate the role of the DNLL in the formation of the plasticity of IC neurons. Here, we used extracellular single unit recording and electrical stimulation to investigate the plasticity of IC neurons in Kunming mice. The results showed that after the cessation of 30-minute electrical stimulation on contralateral DNLL, 95% of the inhibited IC neurons and 86% of the facilitated IC neurons showed plastic changes. Moreover, 1 h after the contralateral DNLL stimulation was stopped, the plastic changes in 74% of the inhibited IC neurons vanished, but still were maintained in 26% of the inhibited IC neurons. These results suggest that the contralateral DNLL ascending input can induce plastic changes of IC neurons, and this kind of effect can be maintained for a certain period of time, which is beneficial to enhance the sound intensity sensitivity of IC neurons.

Application of Chimeric Antigen Receptor-Modified NK Cells in Multiple Myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the killing effect of CAR (CD138-CD28-CD3ζ)-NK cells on myeloma cells through construction of CAR(CD138-CD28-CD3)-NK cells.</p><p><b>METHODS</b>The antiCD138scFv-CD28-CD3 zeta plasmid pcDNA3.1 was constructed, which then together with 3 plasmid lentiviral packaging system were transfected into 293T cells, the virus was collected. Furthermore, in order to get the stably transfected cell line, the NK92MI cell line was infected by the virus, then the positive cells were screened by puromycin. The expression of the CARNK cells were verified by RT-PCR and Western blot. At last the ability of secreting cytokine CD107a was detected by flow cytometry, and the statistical analysis was carried out to verify the anti-myeloma effect of CAR-NK cells.</p><p><b>RESULTS</b>Gene fragment of the CAR(antiCD138scFv-CD28-CD3ζ) was constructed successfully by gene engineering technique in vitro, and the gene sequence was verified to be correct by sequencing. By virus packaging technology, the virus expressing the protein of the CAR was obtained. PCR and Western blot verified the expression of CAR fusion protein on the sufurce of NK cells. The cell killing experiment confirmed that the CAR-NK cells possessed the ability to secrete cytokine CD107a superior to control cells and showed the obvious killing effect on multiple myeloma cells.</p><p><b>CONCLUSION</b>The CAR can be constructed in vitro, and express on NK92 cells. The CAR-NK cells can kill the multiple myeloma cells expressing CD138 antigen, thereby plays an antimyeloma effect.</p>

Effect of rhG-CSF Mobilization on S1P5 Expression in T Lymphocyte Subsets of Allo-HSCT Donors / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) mobilization on S1P5 expression in T lymphocyte subsets of allo-HSCT donors.</p><p><b>METHODS</b>The peripheral blood was collected from 10 allo-hematopoietic stem cell transplantation (allo-HSCT) donors before and after mobilization with rhG-CSF for 4 days. The flow cytometry was used to detect S1P5 expression in T lymphocyte subsets.</p><p><b>RESULTS</b>There was no S1P5 expression on the surface of T-lymphocytes both before and after rhG-CSF mobilization. After fixation with permeabilization agent, S1P5 expression could be detected in lymphocytes after rhG-CSF mobilization, which indicates S1P5 may be located in cells. Compared with level before rhG-CSF mobilization, S1P5 expression was significantly increased in T lymphocyte subsets after rhG-CSF mobilization, CD3(+)T cells (57.92±2.32)% vs (7.94±1.47)%(P<0.05）, CD4(+)T cells (72.58±1.73)% vs （5.48±0.82）%(P<0.05）, CD8(+)T cells（51.79±3.57）% vs （6.46±1.01）%（P<0.05）,CD3-/CD56(+)NK cells（40.00±1.47）% vs（4.97±0.74）%（P<0.05）. The up-regulated level of S1P5 expression in CD4(+)T cells was most high(P<0.05).</p><p><b>CONCLUSION</b>S1P5 expression significantly increases in T lymphocyte subsets after rhG-CSF mobilization, and the up-regulated level of S1P5 expression in CD4(+)T cells is the most high.</p>

Expression Profile of lncRNA NONHSAT040475 in Peripheral Blood Leukocytes of Healthy Persons / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression profile of lncRNA NONHSAT040475 in peripheral blood leukocytes of healthy persons.</p><p><b>METHODS</b>The peripheral blood mononuclear cells were collected from 10 healthy volunteers, the CD3(+) T cells,CD19(+) B cells,CD56(+) NK cells and granulocytes were purified and sorted by flow cytometry. Then, the expression of lncRNA NONHSAT040475 in peripheral blood leukocyte subsets was detected by real-time quantitative PCR.</p><p><b>RESULTS</b>the expression levels of lncRNA NONHSAT040475 were various in lymphocyte subsets, its expression in B lymphocytes was significantly higher, as compared with T lymphocytes, while the expressions of lncRNA NONHSAT040475 in NK cells and granulocytes were relative low（P<0.01）.</p><p><b>CONCLUSION</b>lncRNA NONHSAT040475 is widely expressed in human peripheral blood leukocytes, and mainly expressed in B lymphocytes, therefore, laying the foundation for further study of B lymphocyte-related diseases. This study has brought a new opportunity for diagnosing and illustrating the molecular mechanism of hematologic disorder or autoimmune disease.</p>

Role of Mesenchymal Stem Cells in Preventing GVHD: A Meta-Analysis / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of mesenchymal stem cells (MSC) in the prevention of graft versus host disease (GVHD) after hematopoietic stem cell transplantation (HSCT).</p><p><b>METHODS</b>Randomized controlled trials (RCT) were identified from PubMed (1950.1-2014.3), EMbase (1970.1-2014.3), Cochrane Central Register of Controlled Trials (CENTRAL, issue 4, 2014) of the Cochrane Library, China Biological Medicine (CBM, 1978.1-2014.3). References of retrieved articles were also identified. The quality of each RCT was evaluated by the Cochrane collaboration's tool for assessing the risk of bias. Data analysis was performed with Review Manager 5.1 to evaluate the efficacy of MSC in the prevention of GVHD after HSCT.</p><p><b>RESULTS</b>A total of 3 English articles involving 117 patients were included. Meta-analysis indicated that MSC did not reduce the incidence of acute GVHD and chronic GVHD (RR:0.44, 95% CI: 0.08 to 2.51, P = 0.35; RR:0.85, 95% CI: 0.54 to 1.33, P = 0.47). However, MSC did not increase occurrence of relapse and cytomegalovirus infection (RR:1.52, 95% CI:0.63 to 3.68, P = 0.35;RR:1.05, 95% CI:0.72 to 1.53, P = 0.78). Finally, MSC did not improve overall survival rate of patients received HSCT (RR:1.06, 95% CI:0.79 to 1.43, P = 0.71).</p><p><b>CONCLUSION</b>MSC may have a preventive effect on GVHD in patients undergoing HSCT. However, the evidence is weak due to the small sample sizes. Thus, a reliable conclusion about the preventive effect of MSC on GVHD at the moment has not been made, further larger, high quality, randomized and controlled trials are warranted.</p>

Effects of rhG-CSF mobilization on the polarization and migration of donor's CD4(+)T lymphocytes / 中国实验血液学杂志

The polarization and migration of T lymphocytes involves the adhesive interaction of lymphocyte function-associated antigen 1(LFA-1) with its ligand intercellular adhesion molecule 1 (ICAM-1). This study was aimed to investigate the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) mobilization on the polarization and migration of donor's CD4(+) T cells in peripheral blood. The peripheral blood mononuclear cells were collected from 10 healthy volunteers and 10 donors on the fifth day of mobilization with rhG-CSF. And the CD4(+)T cells were purified by miniMACS. The polarization and migration of CD4(+) T cells activated by stroma cell-derived factor -1α (SDF-1α) and ICAM-1 were detected by using inverted and confocal microscopes respectively. The results showed that the percentage of polarized CD4(+)T cells from donors(32.42 ± 4.91)% was lower than that from healthy controls(56.55 ± 5.35)% (P < 0.01), the migration velocity of CD4(+)T cells from donors (7.06 ± 1.44 µm/min) was also lower than that of healthy controls(9.05 ± 1.91 µm/min)(P < 0.01). It is concluded that the polarization and migration of CD4(+)T lymphocytes is impaired after rhG-CSF mobilization.

Effects of rhG-CSF Stimulation in vitro on the Adhesion and Polarization of Human CD4⁺T Lymphocytes / 中国实验血液学杂志

The adhesion and polarization of T lymphocytes involved in the adhesive interaction of lymphocyte function-associated antigen 1 (LFA-1) with its ligand intercellular adhesion molecule 1 (ICAM-1). This study was aimed to investigate the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) stimulation in vitro on the adhesion and polarization of CD4⁺ T cells of healthy human in peripheral blood. The peripheral blood mononuclear cells were collected from 12 healthy volunteers. The CD4⁺ T cells were sorted by miniMACS. The sorted CD4⁺ T cells were incubated with rhG-CSF for 24 h, then the adhesion and polarization of CD4⁺ T cells activated by stroma cell-derived factor -1α (SDF-1α) and ICAM-1 were detected by ELISA and inverted microscope. The results showed that the percentage of adhesion CD4⁺T cells in the experimental group (rhG-CSF acting on the healthy adult volunteers) (61.9 ± 5.9)% was lower than that in the control group (healthy adult volunteers without rhG-CSF stimulation) (68.3 ± 7.3)% (P < 0.05). The percentage of polarized CD4⁺T cells in the experimental group (24.3 ± 4.3)% was also lower than that in control group (47.1 ± 5.1)% (P < 0.05). It is concluded that the adhesion and polarization of CD4⁺T lymphocytes can be inhibited after rhG-CSF stimulation.