Expressions and clinical significance of GAS1, IL-1RAP and PRF1 in patients with ALK positive anaplastic large cell lymphoma / 中华血液学杂志

Objective: To investigate the expressions of growth arrest-specific protein (GAS1), IL-1 receptor accessory protein (IL-1RAP) and perforin (PRF1) in patients with anaplastic lymphoma kinase positive, anaplastic large cell lymphoma (ALK+ ALCL) and their relationships with clinical significances and the prognoses of ALK+ ALCL. Methods: Twenty-six formalin-fixed paraffin-embedded (FFPE) samples of ALK+ ALCL patients who were diagnosed from January 2011 to September 2016 were collected. Twelve FFPE samples of patients with ALK+ALCL, 13 FFPE samples of patients with peripheral T cell lymphoma (not otherwise specified) (PTCL-NOS) and 8 FFPE samples of patients with angioimmunoblastic T-cell lymphoma (AITL) were used as control groups. RQ-PCR and immunohisto-chemical staining were used to detect the mRNA and protein expressions of GAS1, IL-1RAP and PRF1. The clinical data were analyzed. Results: ①The expression levels of GAS1, IL-1RAP and PRF1 gene and protein in ALK+ ALCL group were higher than those of the control groups (P<0.05), but the expression levels had no statistically significant differences between the control groups (P>0.05). ②Patients with elevated lactate dehydrogenase (LDH) (0.77 vs 1.38, z=-3.292, P=0.001) or International prognostic index (IPI)≥3(0.62 vs 1.29, z=-2.495, P=0.013) had lower expression level of GAS1. Patients with stage Ⅲ/Ⅳ disease (0.89 vs 1.18, z=-2.212, P=0.027) or IPI≥3 (0.48 vs 1.13, z=-2.008, P=0.045) had lower expression level of PRF1. IL-1RAP expression level was not associated with clinical features. ③ALK+ ALCL patients in complete remission (CR) group had higher expression levels of GAS1 and PRF1 than patients in non-remission (NR) group (P values were 0.016 and 0.009). ④Kaplan-Meier survival analysis showed that patients with high expression levels of GAS1 and PRF1 had longer median overall survival and progression-free survival than patients with low expression levels of GAS1 and PRF1. Conclusion: GAS1, IL-1RAP and PRF1 could be molecular markers for ALK+ ALCL patients. They have potential diagnostic value and can be used for differential diagnosis in some difficult cases. ALK+ ALCL patients with high expression levels of GAS1 or PRF1 have better curative effects and prognoses.

Effects of Recombinant Human Erythropoietin on MPC-11 Myeloma Cells Cultured In Vitro / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of recombinant human erythropoietin (rhEPO) on myeloma cell line MPC-11 in vitro and its anti-tumor mechamism so as to provide the theoretic evidence for treatment of multiple myeloma by using rhEPO.</p><p><b>METHODS</b>MPC-11 cells were cultured in RPMI 1640 medium, then the MPC-11 cells in logarithmic growth phase were seed in 96 well culture plates at a density of 1×10/ml cells with 100 µl per well. For all the experiments, the control and rhEPO groups were set up, and the rhEPO group was divided into different concentration groups: 20, 40, 60, 80, 100, 200 and 400 rhEPO U/ml. The cultured MPC-11 cells in each groups were collected at day 1, 2, 3, 4, 5 and 6, and the viability of MPC-11 cells was assayed by using CCK-8 method, the MPC-11 cell apoptosis was detected by flow cytometry with Annexin V/PI double staining, the levels of IL-6, IgG and kappa light chain in cell supernatant were detected by ILISA.</p><p><b>RESULTS</b>The apoptosis rate in MPC-11 cells treated with 200 U/ml of rhEPO at day 21 was significantly higher than that in control group(P<0.05); the apoptosis rate in MPC-11 cells treated with 400 U/ml of rhEPO at day 10, 15, 21 was higher than that in control group at same time points, respectively (P<0.05), suggesting that the MPC-11 cell apoptosis rate was enhanced with increase of rhEPO concentration and prolonging of culture time(P<0.05), the IL-6 level in supernatant of cell treated with 400 U/ml of rhEOP for 15 days was lower than that in control group(P<0.05), while the IL-6 level in supernatant of cells treated with 100, 200, 400 U/ml of rhEPO for 21 days decreased (P<0.05); The levels of IgG and kappa light chain in supernatant of cells treated with 400 U/ml of rhEPO for 21 days all were significantly lower than those in control group(P<0.05).</p><p><b>CONCLUSION</b>The rhEPO can induce apoptosis of MPC-11 cells with concentration- and time-dependent manner, and can reduce levels of IgG, kappa light chain secreted by MPC-11 cells with time-dependent manner.</p>

Diagnostic Value of Protocadherin 15 for NK/T Cell Lymphoma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore whether PCDH15-SI is stable and accurate to diagnose NK/T-cell lymphoma or not.</p><p><b>METHODS</b>The paraffin-biopsiy specimens were collected from 45 cases of NK/T-cell lymphoma and 33 cases of non-NK/T-cell lymphoma in Department of pathology, West China Hospital of Sichuan University. The paraffin sections were stained by immunohistochemistry and blank controls were set up. The peripheral blood was collected from these cases of NK/T-cell lymphoma and non-NK/T-cell lymphoma, as well as from the normal controls in the same hospital. The serum PCDH15 levels were detected by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>Immunohistochemistry showed that PCDH15 were expressed in biopsy specimens of both the NK/T-cell lymphoma in the experimental group and the non-NK/T cell lymphoma in the control group. The expression rate in the NK/T-cell lymphoma group was higher than that in control group. The PCDH15-SI can be detected in NK/T cell lymphoma, non-NK/T cell lymphoma and the normal controls, which is higher in lymphoma patients than that in control, and the serum level was higher in the non-NK/T-cell lymphoma patients than that in the NK/T-cell lymphoma patients. Also, the serum level was higher in the patients with advanced disease than that in the patients at early stage.</p><p><b>CONCLUSION</b>PCDH15 does not act as the new diagnostic marker for NK/T cell lymphoma. The serum level of PCDH15-SI may be helpful to the staging and judging therapeutic effect and prognosis for the patients with non-Hodgkin's lymphoma.</p>

Diagnostic value of serum levels of BamHI-W, LMP-1 and BZLF1 in NK/T-cell lymphoma / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the diagnostic value of serum levels of BamHI-W fragment, latent membrane protein-1 (LMP-1), BZLF1 and ZEBRA protein in patients with natural killer (NK)/T-cell lymphomas (NKTCLs), and to evaluate their relationship with clinical features.</p><p><b>METHODS</b>A total of 144 cases were analyzed in this study, including 48 NKTCLs patients, 48 other types of non-Hodgkin's lymphomas (NHL) patients and 48 healthy individuals as controls. Fluorescent quantitative real-time polymerase chain reaction (RQ-PCR) was used to measure the copy number of BamHI-W, LMP-1 and BZLF1 in serum. Enzyme linked immunosorbent assay (ELISA) was applied to measure the serum levels of ZEBRA protein. The relative operating characteristic (ROC) curve was applied in the evaluation of the tested markers in diagnosis of NKTCL patients, and the correlations among the tested markers and clinical feature were analyzed.</p><p><b>RESULTS</b>Compared with the controls, NKTCL group showed significantly higher levels of all the tested markers (P < 0.01). The median values of serum BamHI-W, LMP-1 and BZLF1 DNAs level were 1870, 394 and 499 copies/ml, respectively. And the median value of ZEBRA protein level was 73.3 µg/L. Furthermore, the ROC curves analysis revealed that all the area under curve (AUC) of LMP-1, BZLF1 and ZEBRA were more than 0.70, which were probably helpful in the diagnosis of NKTCL. To predict the presence of NKTCL, BamHI-W showed a high sensitivity of 81.3%, while BZLF1 showed a high specificity of 81.2%. Untreated patients seemed to have a significantly higher level of serum LMP1 DNA than that of treated patients (median value 898 copies/ml vs 0 copies/ml, P = 0.050). Correlation analysis showed that serum BamHI-W DNA level was correlated with the presence of B symptoms. All the three genes expressed in 94.4% of the untreated cases. On the other hand, none of them expressed in treated cases.</p><p><b>CONCLUSIONS</b>It suggested that combined measurements of BamHI W, LMP1 and BZLF1 DNA levels might be helpful to the diagnosis and therapeutic monitor of NKTCL.</p>

The clinical characteristics of 681 patients with diffuse large B-cell lymphoma / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze clinical features and the differences of GCB and non-GCB phenotypes for diffuse large B-cell lymphoma (DLBCL) in different age groups, Ki-67 index and international prognostic index (IPI).</p><p><b>METHODS</b>Clinical data of 681 patients with DLBCL hospitalized in West China Hospital from January 2000 to December 2010 were retrospectively analyzed.</p><p><b>RESULTS</b>Of these DLBCL cases, the median age was 56 years old with a male predominance, 51.4% stage III-IV, 37.6% B symptoms, 30.2% IPI 3-5 scores, 49.8% from extranodal sites, 29.0% gastrointestinal tract infiltration, 38.3% low absolute lymphocyte count (ALC), 56.1% elevated serum lactate dehydrogenase (LDH) level, 83.0% elevated β(2)-microglobulin (β(2)-MG) level. B symptoms was associated with bone marrow involvement with the odds ratio 5.212 (95%CI 2.821 - 9.632, P = 0.000). Among 268 with DLBCL patients classified by Hans' classification, 28.4% were GCB and 71.6% non-GCB. The proportions of patients with HBsAg-positive, elevated serum LDH level and Bcl-2 positive expression in non-GCB group was higher than those in GCB group (P < 0.05). The differences between GCB and non-GCB DLBCL were not revealed in terms of age subgroups, Ki-67 expression status and IPI subgroups. The high (≥ 60%) Ki-67 group included more patients with extranodal site involvement compared with the low (< 60%) Ki-67 group (51.8% vs 38.7%, P = 0.008). The proportion of patents with low ALC in IPI 3-5 scores group was higher than in IPI 0-2 scores group (P = 0.000). The multivariate analysis showed that high IPI had statistically significant negative influence on survival (P = 0.000).</p><p><b>CONCLUSIONS</b>Most patients with DLBCL were middle-aged male from our data. The patients with primary nodal (PN) was almost equal to those with primary extranodal (PEN). The most frequent extranodal site was gastrointestinal tract. The non-GCB phenotype was significantly more common than GCB phenotype in this study, and the non-GCB group included more patients with HBsAg-positive and Bcl-2 positive expression. Low ALC was observed predominantly in the high risk group. IPI score was an independent prognostic indicator for survival.</p>

The clinical features, therapeutic effects and prognosis of mantle cell lymphoma / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze clinical features, therapeutic effects and prognostic factors of patients with mantle cell lymphoma (MCL).</p><p><b>METHODS</b>Clinical data of 37 MCL patients hospitalized in our hospital from January 2000 to March 2010 were retrospectively analyzed.</p><p><b>RESULTS</b>The median age was 62, with a male predominance. 97.30% of the patients were in Ann Arbor stage III ∼ IV, 54.05% with B symptoms, 64.86% with bone marrow involvement, 29.73% with splenomegaly, 24.32% with lymphocytosis and 51.35% with elevated LDH. Ki-67 was detected in 22 cases, and patients with Ki-67 ≤ 40% accounted for 68.18%. Of 37 cases, the overall response rate (ORR) of rituximab combined with chemotherapy was 92.31%, being higher than those of CHOP (46.15%) and CHOP + IFN (42.86%) regimens. There were statistical differences in the 3-year progression-free survival (PFS) and overall survival (OS) between rituximab + chemotherapy and CHOP or CHOP + interferon regimens (P < 0.05, respectively). Splenomegaly, elevated WBC, lymphocytosis and Ki-67 > 40% were identified as adverse prognostic factors.</p><p><b>CONCLUSION</b>Most patients with MCL were older adults, with a male predominance and usually had bone marrow involvement and poor prognosis. Rituximab combined with chemotherapy could improve ORR and OS of MCL.</p>

Establishment of a multiple myeloma local tumor model in mice / 中国实验血液学杂志

This study was purposed to establish a multiple myeloma local tumor model in the BLAB/c mice. Healthy BLAB/c mice were injected subcutaneously with 6 x 10(5) MPC-11 cells. In the peak time of the subcutaneous nodules observed, five mice were randomized selected to be executed and the subcutaneous nodules of these mice executed were used to detect the CD138 and kappa light chain by means of HE staining and the immunohistochemistry methods. The serum immunofixation electrophoresis (IFE) of tumor-bearing mice were performed at 5, 7, 9, 11, 12, 35 and 65 days after the initial MPC-11 cell injection. Hemoglobin level was assayed at 15 and 30 days after the initial MPC-11 cell injection. The serum levels of IL-6 were also assayed at 35 and 65 days after the initial MPC-11 cell injection. The tumor volume was monitored twice a week and their body weights were measured once a week. The results showed that the peak of the subcutaneous nodules appeared at 12 to 15 days after the initial MPC-11 cell injection. The serum monoclonal immunoglobulin could be detected at 12 days after MPC-11 cell injection. The results of HE staining and immuno-histochemistry assay for detection of CD138 and kappa light chain positive expressions proved that the subcutaneous tumor nodules originated from MPC-11 plasmacytes. The serum monoclonal protein (M protein) of the tumor-bearing mice was detected at 12 days after bearing tumor which manifested thick bands of IgG and kappa light chain. The peak time of mortality was at 20 to 40 days after the initial MPC-11 cell injection, and the median survival time was 31 days. Anemia in mice appeared at 15 days. There was a significant difference of Hb level between the tumor-bearing group and the normal group at 15 and 30 days respectively (p < 0.05). The serum level of IL-6 in tumor-bearing mice was higher than that in the normal group. It is concluded that to establish the multiple myeloma local tumor model in mice by using subcutaneous injection of MPC-11 cells has various advantages, such as simple method of model established, relative high success of bearing tumor, easy observation of tumor growth change and so on. This model can be useful for studying and evaluating the therapeutic efficacy for multiple myeloma through monitoring the changes of tumor size, serum IL-6 level and serum immunofixation electrophoresis.

Analysis of T cell subsets and CD3zeta chain expression in myelodysplastic syndrome patients / 中国实验血液学杂志

Immune mediated suppression of hematopoiesis has been considered as one of the most important mechanisms leading to pancytopenia in myelodysplastic syndromes. This research was aimed at evaluating immune state of the MDS patients, analyzing the peripheral blood T cell subsets and CD3zeta chain expression and searching the possible reasons of hematopoietic disorders in 11 cases of MDS. Peripheral blood mononuclear cells were collected from 11 patients whose diagnosis was confirmed according to the new WHO diagnostic criteria. Flow cytometry was used for the counts of IFNgamma(+)CD4(+) cell (Th1), IL4(+)CD4(+) cell (Th2), IFNgamma(+)CD8(+) cell (Tc1), and IL4(+)CD8(+) cell (Tc2), and for the analysis of expression of CD3zeta chain in T cell subsets. The results showed that CD8(+) cells increased significantly in MDS patients; there was no significant difference between Th1/Th2, Tc1/Tc2 ratios of T cell subsets and normal control; CD3zeta chain, the functional protein in the signal transduction pathway of T cell, was over expressed in the CD8(+) cell. In conclusion, research indicates that abnormal changes of T cell subgroups exist in peripheral blood of MDS patients. Enhancement of CD8(+) cells and over-expression of CD3zeta chain are important features, which suggest that CD8(+) cells play the most critical role in the pathologic process as compared with other T cell subsets. The over active immunity mediated by T cell subset may be one of the major mechanisms resulting in cytopenia in MDS.

A case-control study for assessing the relation between the incidence of malignant lymphomas and environmental factors in Sichuan province / 中华流行病学杂志

<p><b>OBJECTIVE</b>To evaluate the relationships of malignant lymphoma (ML) and exposure factors, including environmental, drug using, socioeconomic level and life style etc.</p><p><b>METHODS</b>(1) Hospital-based case-control study with 1:2 matching was used; (2) 150 cases of ML and 300 controls were resulted; (3) logistic regression was used.</p><p><b>RESULTS</b>(1) Adjust odds ratios (aOR) regarding contact to organic solvents were: benzene aOR = 2.78, P = 0.001;banana oil aOR = 2.28, P = 0.043; paint aOR = 1.96, P = 0.023; (2) aOR on pesticides contact: organic phosphorus aOR = 1.98, P = 0.034; chrysanthester aOR = 2.57, P = 0.013; organic nitride aOR = 2.55, P = 0.003; (3) aOR of using aspirin was 2.36, P = 0.005; (4) dog-raising: aOR = 1.76, P = 0.034; (5) the aORs of interval-term exposure to small dosage X-ray was 0.49, P = 0.015, and frequently catching cold was 0.67, P = 0.024; (6) the P-value of different occupations was more than 0.05; cigarette smoking OR = 1.01, P = 0.897; giving up smoking OR = 1.17, P = 0.073; alcohol-drinking aOR = 1.01, P = 0.414; hair-dyes OR = 1.06, P = 0.850; hair-perms OR = 1.26, P = 0.438; socioeconomic level OR = 1.00, P = 1.000 and with introversive character OR = 1.26, P = 0.266.</p><p><b>CONCLUSIONS</b>(1) Factors that might increase the risk of suffering from malignant lymphomas would include direct contact to organic solvents and some pesticides (organic phosphorus, chrysanthester and organic nitride), intake of aspirin, as well as dog-raising. (2) Regular exposure to small dosage X-ray and frequently catching cold seemed to decrease the risk of suffering from ML. (3) Occupation, smoking, giving up smoking, alcohol-drinking, hair-dyes, hair-perms, socioeconomic level and introversive character showed no significant relations to the risk of suffering from ML in this study.</p>