[Expression and Clinical Significance of Cytokines and Lymphocyte Subsets in Patients with Diffuse Large B-Cell Lymphoma].

OBJECTIVE: To study the role of cytokines and lymphocyte subsets in the diagnosis, prognosis and efficacy evaluation of DLBCL patients, and the effects of Tislelizumab on immune function and cytokines in DLBCL patients. METHODS: Twenty-three patients with newly diagnosed DLBCL were selected as DLBCL group and 34 patients with megaloblastic anemia as the control group. The levels of peripheral blood cytokines IL-2, IL-4, IL-6, IL-10, TNF- α and IFN-γ by ELISA method. The levels of peripheral blood CD3+, CD4+, CD8+ T lymphocytes, B lymphocytes and NK cells， the ratio of CD4+/CD8+ were detected by flow cytometry. The levels of cytokins and lymphocyto subsets in DLBCL patients with different clinical data and different therapeutic effects were compared. RESULTS: The levels of cytokines IL-2, IL-6 and IL-10 in DLBCL group were significantly higher than those in control group, but there was no significant correlation between cytokine levels and age and gender. The higher IPI score, higher Ann Arbor stage, B symptoms, higher ß 2-MG, LDH and CRP levels, IL-6 and IL-10 levels were significantly higher, and IL-4 was also significantly higher in patients with high LDH levels. Compared with the ineffective group, the levels of IL-6 and IL-10 were significantly lower and the level of CD4+ T cells and the ratio of CD4+/CD8+ was significantly higher in the effective group before therapy. The levels of IL-6, IL-10 and B lymphocytes in the effective group decreased significantly after therapy compared to those before therapy. After 4 cycles of therapy, the level of IL-2 and the ratio of CD4+/CD8+ in the Tislelizumab group were significantly higher than those in the non-Tislelizumab group, and the level of CD8+ T cells was significantly lower than that in the non-Tislelizumab group(P<0.05). The level of B lymphocytes in both the Tislelizumab group and the non-Tislelizumab group after therapy was significantly lower than that before therapy. CONCLUSION: The expression of cytokines and lymphocyte subsets in peripheral blood of patients with DLBCL is abnormal, which is related to the severity, prognosis and therapeutic effect of the disease. Tislelizumab can improve the immune function of patients with DLBCL by affecting cytokines and lymphocyte subsets and strengthen anti-tumor immunity.

Coexistence of diffuse large B-cell lymphoma, acute myeloid leukemia, and untreated lymphoplasmacytic lymphoma/waldenström macroglobulinemia in a same patient: A case report.

BACKGROUND: The Coexistence of myeloid and lymphoid malignancies is rare. Myeloid leukemia occurs more frequently as a secondary event in patients receiving chemotherapy agents for lymphoid malignancies. Synchronous diagnoses of diffuse large B-cell lymphoma (DLBCL), acute myeloid leukemia (AML), and untreated lymphoplasmacytic lymphoma/Waldenström macroglobulinemia (LPL/WM) in the same patient have not been reported. Here we report one such case. CASE SUMMARY: An 89-year-old man had a chest wall mass histopathologically diagnosed as DLBCL. The bone marrow and peripheral blood contained two groups of cells. One group of cells fulfilled the criteria of AML, and the other revealed the features of small B lymphocytic proliferative disorder, which we considered LPL/WM. Multiple chromosomal or genetic changes were detected in bone marrow mononuclear cells, including ATM deletion, CCND1 amplification, mutations of MYD88 (L265P) and TP53, WT1 overexpression, and fusion gene of BIRC2-ARAP1, as well as complex chromosomal abnormalities. The patient refused chemotherapy because of old age and died of pneumonia 1 mo after the final diagnosis. CONCLUSION: The coexistence of DLBCL, AML, and untreated LPL/WM in the same patient is extremely rare, which probably results from multiple steps of genetic abnormalities. Asymptomatic LPL/WM might have occurred first, then myelodysplastic syndrome-related AML developed, and finally aggressive DLBCL arose. Therefore, medical staff should pay attention to this rare phenomenon to avoid misdiagnoses.

[Expression Level of S100A6 mRNA in MM and Its Clinical Significance].

OBJECTIVE: To investigate the expression level of S100A6 mRNA in MM and to its clinical significance, and to evaluate its significance. METHODS: The expression level of S100A6 mRNA in MM patients was determined by real time quantitative PCR(RQ-PCR), and its relationship with the clinical features and outcomes of patients was analyzed by statistic method. RESULTS: S100A6 mRNA was detected in 20 MM patients. Compared with normal persons, the S100A6 mRNA expression in MM patients was higher. In different groups, the S100A6 mRNA expression in MM patients of 3 stages was higer than that in patients of 1 and 2 stages. MM patients with higher S100A6 mRNA expression had poor prognosis and higer extramedullary metastasis rate. CONCLUSION: The high expression of S100A6 mRNA is associated with poor prognosis and may be a prognostic molecular marker of MM.

[Effect of free light chain ratio normalization after treatment on prognosis of patients with multiple myeloma].

This study was aimed to investigate the normalization of serum free light chain ratio (sPLCR) after treatment of multiple myeloma (MM) and its influence on the prognosis of MM patients. The clinical data of 42 patients with MM were analyzed retrospectively from January 2009 to November 2013 in out department. According to sPLCR consecutive normalization for more 4 weeks or not after treatment, the patients were classified in patients with mormalized sPLCR and patients with abnormalized sPLCR, then the influence of traditional prognostic factors of MM on sPLCR and effect of sPLCR on overall survival (OS) time of MM patients were analyzed. The results showed that the influence of age, ISS stage displayed statistical difference between sFLCR normalization group and abnormalization group, the age ≥ 65 years and ISS stage III negatively impacted on sFLCR normalization (P < 0.05). The response rates of patients with normalized sFLCR were as follows: CR - 60%, VGPR - 38.89%; PR - 28.57%; 17 patients (40.48%) with sFLCR normalization showed superior OS, as compared with patients with sPLCR abnormalization (P < 0.01). It is concluded that the sFLCR normalization is the independent prognostic factor for MM, suggesting that the MM patients with sPLCR normalization after treatment have superior prognosis.

[Myeloid/natural killer cell acute leukemia resembling acute promyelocytic leukemia].

In order to improve the recognition of myeloid/natural killer cell acute leukemia and to reduce misdiagnosis, one case of myeloid/natural killer cell acute leukemia resembling acute promyelocytic leukemia(APL) was reported and the related articles published were reviewed. A series of clinical tests, the morphologic and immunophenotypic analysis of leukemia cells, cytogenetic and molecular biological examinations were performed. The results indicated that the patient had anemia, thrombocytopenia and leucocytosis, but no evidence of lymphadenopathy and hepatosplenomegaly. The morphology of leukemia cells was similar to that of abnormal promyelocytic cells, especially the variant of M3 (M3v) leukemia cells. The leukemia cells expressed CD117, CD33, CD15, CD56 and cMPO, but did not express CD34, HLA-DR, CD13 and CD16. Abnormal cytogenetics with del (7) (q22q32) was found. Neither t(15;17) nor PML/RARα gene rearrangement was detected. The patient failed to show a differentiation-induction response to all-trans retinoic acid(ATRA). In conclusion, the myeloid/natural killer cell leukemia is extremely rare. It is very important to distinguish the disorder from APL/M3v. The patient with myeloid/natural kill cell acute leukemia should be treated with chemotherapy as acute myeloid leukemia.

[Apoptosis-inducing effect of annexin A2 on multiple myeloma cells and its related mechanisms].

This study was purposed to investigate the apoptosis-inducing effect of Annexin A2 gene (AnxA2) on multiple myeloma (MM) cells and its mechanisms. The human MM cell lines U266 and RPMI8226 were transfected by using siRNA targeting at AnxA2; the expressions of AnxA2 mRNA and protein in the siRNA-transfected cells were detected by real-time PCR and Western blot, respectively; the cell apoptosis was assayed by flow cytometry. The results showed that silencing AnxA2 gene by siRNA resulted in decreased expressions of AnxA2 gene and protein, increased apoptosis of U266 and RPMI8226 cell lines (P < 0.05), at the same time resulted in down-regulation of apoptosis-related gene expressions including p65NF-κB, IL-2, IL-6 (P < 0.05), and up-regulation of P53 gene expression (P < 0.05). It is concluded that the AnxA2 silence plays a promoting role in apoptosis of MM cell lines U266 and RPMI8226.

[Angioimmunoblastic T-cell lymphoma with large granular lymphocytosis].

To improve the recognition of angioimmunoblastic T-cell lymphoma (AITL) and to reduce misdiagnosis, a case diagnosed as AITL with large granular lymphocytosis was reported and the related articles were reviewed. A series of clinical tests, pathologic examination and immunohistochemical test, TCR gene rearrangement detection by multiple PCR and assay of lymphocyte immunophenotypes were carried out. The results indicated that the patient was characterized by fever, skin rash, generalized lymphadenopathy, splenomegaly, pleural effusion, ascites, anemia and thrombocytopenia, increase of circulating large granular lymphocytes with CD3(-) and CD16(+), CD56(+) were detected, T-cell receptor γ-chain gene was rearranged. More large granular lymphocytes with abnormal mitosis were found in ascites. The histological and immunohistochemical changes observed by the lymph node biopsy were compatible with AITL, some cells of which were CD56-positive. In conclusion, AITL is characterized by aggressive progress and generally occurs in elderly patients, its clinical prognosis is poor, the large granular lymphocytosis may be an autoimmune response to the tumor cells or originate from tumor stem/progenitor cells.

[Effects of thalidomide on Annexin II gene regulation].

OBJECTIVE: To investigate the effect of thalidomide on Annexin II (AnxA2) gene regulation in multiple myeloma cell line RPMI8226 and human microvascular endothelial cell line HMEC-1 cells in vitro, and explore the potential mechanism of thrombosis induced by thalidomide. METHODS: RPMI8226 and HMEC-1 cells were cultivated in vitro. Real time quantitative PCR (RQ-PCR) was used to detect the influence of thalidomide at different concentration on the expression of AnxA2 mRNA, flow cytometry (FCM) and confocal microscopy were used to detect the cell surface protein level after the samples were stimulated with different concentrations of thalidomide. RESULTS: AnxA2 mRNA level in RPMI8226 cells treated with thalidomide at 12.5 microg/ml, 25.0 microg/ml and 50.0 microg/ml was decreased compared with the control group (0.60+/-0.15, 0.33+/-0.14, 0.42+/-0.16, vs 1.07+/-0.16, respectively, P<0.05) and did so in HMEC-1 cells (0.21+/-0.20, 0.08+/-0.08, 0.17+/-0.16 vs 1.16+/-0.24, respectively, P<0.05). The AnxA2 protein level in RPMI8226 cells treated with above mentioned concentrations of thalidomide was also decreased compared with the control (3.39+/-0.32, 2.82+/-0.28, 3.21+/-0.23 vs 5.53+/-0.32, respectively, P<0.05) and that did so in HMEC-1 cells (0.72+/-0.11, 0.64+/-0.08, 0.67+/-0.08 vs 1.40+/-0.15, respectively, P<0.05). CONCLUSIONS: Thalidomide can inhibit the expression of AnxA2 mRNA and protein in RPMI8226 and HMEC-1 cells, which may be one of the mechanisms for the development of thrombosis induced by thalidomide in multiple myeloma patients.

[Effects of Annexin II gene silencing by siRNA on proliferation and invasive potential of Jurkat lymphoma cells].

OBJECTIVE: To investigate the effects of Annexin II (AnxA2) gene silencing by siRNA on proliferation and invasive potential of lymphoma cell line Jurkat cells. METHODS: A synthesized siRNA duplex targeting to AnxA2 was transfected into Jurkat cells. Transfection efficiency was analyzed by real-time PCR and flow cytometry. MTT assay for cell proliferation and transwell plates for invasive potential were performed. RESULTS: Compared with the negative controls, the cell proliferation inhibitory rate of the AnxA2 siRNA transfected Jurkat cells was significantly increased at 24 h, 48 h and 72 h [(17.4 +/- 2.3)%, (22.4 +/- 3.8)%, (37.6 +/- 1.5)% vs (-1.3 +/- 5.1)%, (-5.5 +/- 4.4)%, (-10.8 +/- 5.5)%, respectively, P<0.05]. The cell invasive potential of the transfected Jurkat cells was inhibited remarkably at 48 h (11.3 +/- 4.2 vs 54.3 +/- 8.7, P<0.01). CONCLUSION: AnxA2 gene silenced by siRNA can inhibit the proliferation and the invasive potential of Jurkat cells remarkably.

[Expressions of vascular endothelial growth factor and cyclooxygenase-2 in patients with multiple myeloma and its significance].

This study was aimed to investigate the expressions of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) in patients with multiple myeloma (MM) and its clinical significance. Expression of VEGF was detected by enzyme linked immunosorbent assay (ELISA) and the level of COX-2 was detected by Western blot. The results showed that the serum VEGF level of multiple myeloma patients (365.34 +/- 65.63 pg/ml) was higher than that in the normal persons (122.52 +/- 39.29 pg/ml) (p < 0.05); the serum VEGF level of patients at advanced stage (395.07 +/- 54.90) pg/ml was higher than those at stable stage (300.33 +/- 44.22) pg/ml (p < 0.05). The serum Cox-2 positive rate in the patients (31%) was higher than that in normal persons (0%) (p < 0.01); the serum Cox-2 positive rate in the patients at advanced stage (50%) was higher than those at stable stage (21%) (p < 0.01). It is concluded that VEGF and COX-2 may play an important role in the pathogenesis and development of multiple myeloma, they can be used to evaluate the status of patients with MM.