[The Reduction of CD4+ T Lymphocytes after the Treatment of Follicular Lymphoma with the Bendamustine Containing Regimen May Predict the Occurrence of Infection and Efficacy].

OBJECTIVE: To investigate the effectiveness, safety, and related prognostic factors of the treatment of follicular lymphoma (FL) with a regimen containing Bendamustine. METHODS: The clinical data of 129 FL patients who were treated with Bendamustine containing regimen were collected from January 1,2020 to October 30,2022 in the Hematology Department of Lianyungang Second People's Hospital and Jiangsu Provincial People's Hospital. The patients were divided into three groups: Bendamustine plus Rituximab (BR), Bendamustine plus Obinutuzumab (GB), Rituximab + Cyclophosphamide + Epirubicin / Doxorubicin + Vindesine + Prednisone (R-CHOP). The efficacy, safety and related prognostic factors of the treatment of FL with a regimen based on Bendamustine were retrospectively analyzed. RESULTS: The ORR was 98% for the BR group, 94% for the GB group, and 72.3% for the R-CHOP group, while the CR rate was 61.2%,70% and 40.4%, respectively. The ORR and CR rates of the R-CHOP group were statistically different from those of the BR group and GB group (P < 0.05). The 3-year PFS rate of the BR group, GB group, and R-CHOP group was 89.6%, 90.9%, 48.9%, respectively. There was a statistically significant difference in 3-year PFS between the R-CHOP group, BR group, and GB group (P < 0.05), while there was no statistically significant difference in 3-year OS（P ＞0.05）. Hematological adverse reactions were mainly bone marrow suppression. Lymphocytes and CD4+T lymphocytes decreased to the lowest level about 6 months after treatment, and the incidence of lymphopenia in BR group and GB group was higher than that in R-CHOP group, with a statistical difference (P < 0.05). The higher incidence of non-Hematological adverse reactions were pulmonary infection, EB virus infection, hepatitis B virus reactivation, and gastrointestinal reactions without statistical difference in 3 groups (P >0.05), and were all controllable. The Receiver operating characteristic of CD4+T lymphocyte count showed that AUC of BR group was 0.802, and the critical value was 258/uL; AUC of GB group was 0.754 with a critical value of 322/uL. CONCLUSION: The treatment of FL with the Bendamustine containing regimen has good efficacy and controllable adverse reactions, but lymphocytopenia was significant after treatment, and the curative efficacy in combination with various CD20 monoclonal antibodies was different. The lowest CD4+T lymphocyte count can be used as a predictive factor for the occurrence of infection and efficacy of the Bendamustine containing regimen for FL.

[Clinical significance of tissue factor and vascular endothelial growth factor expressions on CD14+ monocytes in patients with non Hodgkin lymphoma].

OBJECTIVE: To explore the values of tissue factor (TF) and vascular endothelial growth factor (VEGF) expressions on peripheral CD14+ monocytes in disease assessment, prognosis, and short-term efficacy evaluation of non-Hodgkin lymphoma (NHL) patients. METHODS: TF and VEGF expressions on CD14+monocytes in 47 NHL patients (disease group) before chemotherapy and after 4 chemotherapy cycles and in 30 healthy subjects (control group) were detected by flow cytometry, and the potential relationship among TF, VEGF, International Prognostic Index (IPI), and short-term efficacy were analyzed. RESULTS: TF and VEGF expressions on CD14 + monocytes in disease group were significantly higher than those in control group ( all P <0. 01) and positive correlation was showed between them (r = 0. 708, P = 0.00). TF and VEGF expressions in Ann Arbor stage III and IV (n = 22 and 19) , symptomatic (n = 22) , lactate dehydrogenase (LDH) increased (n = 21) , Eastern Cooperative Oncology Group (ECOG) score 2-4 (n = 12) and extranodal lesions >1 (n = 16) groups were significantly higher than those in Ann Arbor stage II (an = 6) , asymptomatic (an =25) , LDH normal (n = 26) , ECOG score 0-1 ( n = 35) and extranodal lesions ~1 ( na = 31) groups, respectively (all P <0.05). The expressions of TF and VEGF on CD14 + monocytes in high-risk (n = 7) or high-middle-risk (n = 11) groups were significantly increased compared with low-risk (n = 15) or low-middle-risk(n = 14) groups, respectively (all P <0. 01). TF and VEGF expressions in non-remission group before chemotherapy (n = 11) were both obviously higher than those in remission group (an = 36, all P <0. 01) , and after chemotherapy their expressions in remission group were significantly lower than those before chemotherapy (all P <0. 01) , while such significant changes were not observed in the non-remission group ( all P > 0. 05). CONCLUSION: The high expressions of TF and VEGF on peripheral CD14 + monocytes can be useful markers in dis-ease assessment, prognosis evaluation and short-term efficacy observation of NHL patients.

[Prognostic Significance of CUEDC1 in Patients with Acute Myeloid Leukemia (non-M3)].

OBJECTIVE: To investigate the prognostic significance of CUEDC1 in patients with acute myeloid leukemia (non-M3). METHODS: 52 cases newly diagnosed AML (non-M3) were selected and enrolled in AML non-M3 group, at the same time, 10 cases of iron doficiency anemia were selected and enrolled in control group. The bone marrow mononuclear cells(BMMC) were isolated from bone marrow of patients, the expression level of CUEDC1 in BMMC was detected by RT-PCR, the expression level of CUEDC1 mRNA in BMMC of AML-subtype patients was compared. The AML patients were divided into low and high expression groups according to the expression level of CUEDC1 mRNA, and the complete remission rate after the first chemothrapy course was compared, and the relative expression level of CUEDC1 mRNA between the remission and the non-remission group were compared. RESULTS: CUEDC1 was expressed in BMMC of 52 newly diagnosed patients with AML (non-M3) of all subtypes, which was higher than that in control group (P<0.05), and the expression level of CUEDC1 mRNA in M5 patients was the highest (P<0.05). In CUEDC1 low expression group, induced complete remisson rate （76.2%,16/21） after the first course of treatment seemed higher than that of the high expression group（67.7%,21/31）, but the difference was not statistically significant; the expression level of CUEDC1 mRNA in the remission group of patients with newly diagnosed AML(non-M3) was lower than that in the non-remission group(P<0.05). CONCLUSION: CUEDC1 is highly expressed in newly diagnosed patients with AML, among which the CUEDC1 mRNA expression level in M5 patients is the highest, the expression of CUEDC1 mRNA possibly relates to the prognosis of patients with AML.

[Clinical Significance of TF and VEGF Expressions on Peripheral CD14 Positive Monocytes in Patients with Diffuse Large B Cell Lymphoma].

OBJECTIVE: To investigate the clinical significance of tissue factor (TF) and vascular endothelial growth factor (VEGF) expression on peripheral blood CD14 positive monocytes in patients with diffuse large B cell lymphoma (DLBCL). METHODS: The expressions of TF and VEGF on peripheral CD14+ monocytes in 41 patients with DLBCL (DLBCL group) before chemotherapy and after 4 chemotherapeutic courses, and in 20 healthy subjects (control group) were detected by flow cytometry respectively, meanwhile, the relationship of the expression of TF and VEGF with international prognostic indexes (IPI) and short-term effects were analysed. RESULTS: The expression levels of TF and VEGF on peripheral CD14+ monocytes in DLBCL group were significantly higher than those in control group (P<0.01), and a positive correlation was found between the two groups (r=0.755, P<0.01). The expression of TF and VEGF on CD14+ monocytes in patients with prognostic risk factors significantly increased as compared with those in patients without prognostic risk factors (P<0.05), but there were no significant differences of TF and VEGF expressions on CD14+ monocytes in DLBCL group with different sex, age, subtypes (P＞0.05). As compared with patients without prognostic risk factors, the expression levels of TF and VEGF on CD14+ monocytes of patients with prognostic risk factors significantly increased (P<0.05). The expression of TF and VEGF on CD14+ monocytes in DLBCL group showed an increasing tendency along with the increase of IPI index (P<0.01). The expression levels of TF and VEGF on CD14+ monocytes in remission group before chemotherapy were lower than those in non-remission group (P<0.01); after chemotherapy, the expression levels of TF and VEGF on CD14+ monocytes in remission group were lower than those before chemotherapy (P<0.05), while the TF and VEGF expression levels in non-remission group were no singnificauly different from TF and VEGF levels before chemtherapy (P>0.05), the survival of patients in group with low expression of TF and VEGF was superior to that in group with high expression of TF and VEGF (P<0.05). CONCLUSION: The paripheral blood CD14+ monocytes in DLBCL patients highly express the TF and VEGF, which relate with IPI, therapeutic efficacy and survival, thus the TF and VEGF expression levels are of reference significance for evaluating the therapeutic efficacy and prognosis of patients.

[Construction of A Lentiviral Vector Carrying CUEDC1 Gene and Its Effect on the Proliferation and Colony-formating Ability of MOLT-4 Cells].

OBJECTIVE: To construct a lentiviral vector carrying human CUEDC1 gene, to establish leukemic cell line MOLT-4 stably expressing recombinant plasmid, to analyze the expression of CUEDC1 in MOLT-4 cells and to investigate its effect on the proliferation of MOLT-4 cells. METHODS: The CUEDC1 gene was amplified by RT-PCR, and then was subcloned into the lentiviral vector pCDH to generate a lentiviral vector pCDH-CUEDC1. Recombinant lentivirus was generated by co-transfection of 3 plasmids, and transfected into MOLT-4 cells. The Real-time PCR and Western blot were respectively applied to detect the expression of CUEDC1 mRNA and protein, the CCK-8 and colony formation assay were used to evaluate the effect of CUEDC1 on proliferation of MOLT-4 cells. RESULTS: The recombinant lentiviral vector pCDH-CUEDC1 had been constructed successfully. After infection of MOLT-4 cells with the lentivirus, the recombinant plasmid could stably up-regulate the expression of CUEDC1 and protein. The CCK-8 detection and colony formation assay showed that exogenous CUEDC1 could significantly promote cell growth and the colony formation of MOLT-4 cells. CONCLUSION: The recombinant lentiviral vector carrying human CUEDC1 has been successfully constructed, exogenous CUEDC1 can significantly promote cell growth and the colony formation of MOLT-4 cells.

[Effect of OCT4A Gene on the Biological Characteristics of K562 Cells].

OBJECTIVE: To evaluate biological effects of OCT4A gene on K562 cells and explore the molecular mechanism of K562 cell apoptosis. METHODS: Two recombinant lentiviral vectors were constructed, which could stablely up- regulate and down- regulate OCT4A protein. Recombinant lentivirus was generated by co-transfection of three-plasmids and transfec-ted into K562 cells. The experiments were divided into 5 groups: normal, pLVX-OCT4A-ZsGreen1, pLVX vector control, PLB-OCT4A shRNA and non-specific shRNA groups. Western blot was applied to detect the expression of OCT4A protein, the cell counting kit-8 was applied to evaluate the effect of OCT4A on proliferation of K562 cells. The apoptosis and differentiation of K562 cells were detected by flow cytometry with AnnexinV/7-AAD double staining. The mRNA expressions of caspase-3,BIM,BCL-xL,BAX in K562 cells were determined by real time PCR. RESULTS: The OCT4A fragment was amplified by reverse transcription polymerase chain reaction(RT-PCR), the 2 lentiviral vectors were successfully constructed. In comparson with those in the control group, the expression of OCT4A protein of pLVX-OCT4A-ZsGreen1 group was significantly increased, but decreased in PLB-OCT4A shRNA group. CCK-8 assay showed that the higher the content of OCT4A protein, the faster the cell proliferation. The apoptosis rate was (3.48±0.52)% of pLVX-OCT4A-ZsGreen1 group, which was lower than that of control group, while the apoptosis rate PLB-OCT4A shRNA group was (7.25±0.57)%, which was higher than that of control group (P<0.05), however, the K562 cells differentiation was not influenced(P>0.05). Compared with control group, the gene expression of Caspase-3,BIM and BAX was down-regulated(P>0.05), but a significant up-regulation of BCL-xL gene expression was observed(P<0.05). CONCLUSION: Two lentiviral vectors have been successfully constructed, which can stably up- and down- regulate the expression of OCT4A in K562 cells respectively. OCT4A can promote the K562 cell proliferation and inhibit the apoptosis, the mechanism may be related with up-regulation of BCL-xl expression.

[Clinical and Pathologic Features of Myeloid Sarcoma].

OBJECTIVE: To explore the clinicopathologic features, differential diagnosis and therapy of myeloid sarcoma. METHODS: The clinical data including clinical manifestations, laboratorial tests, histopathologicical examination, immunohistochemistry and clinical prognosis of 10 patients with myeloid sarcoma were analyzed retrospectively. Among 10 patients, 5 male and 5 female, aged 23 to 71 years old (median = 36 years). RESULTS: 2 cases of myeloid sarcoma were secondary from chronic myeloid leukemia, and 1 cases of myeloid sarcoma occurred after the allogeneic hematopoietic stem cell transplantation due to acute myeloid leukemia, and the others lacked the anamnesis of malignancies. The neoplasms occurred at bone, brain, skin, breast, epididymis, uterine cervix, small intestine, ovary and lymph nodes. Microscopically, the tumor cells were round or oval, which infiltrated diffusely or arranged in single-file. The cytoplasm was scarce and immature eosinophils were scattered. The nuclei were round, oval or focally irregular, and the mitosis was visible. The neoplasms were positive for MPO, CD34, CD43, CD45, CD99 and CD117 by immunohistochemical staining. 4 patients progressed into acute myeloid leukemia from 2 to 10 months after the diagnosis of myeloid sarcoma. All of them achieved complete remission after inductive chemotherapy, but 3 patients relapsed from 3 to 12 months after remission and only survived for 14 to 23 months. 4 patients were treated by using chemotherapy before bone marrow abnormality, and with the disease-free survival for 1 to 48 months. CONCLUSION: Myeloid sarcoma needs to be distinguished from lymphoblastic lymphoma, Burkitt's lymphoma, blastic plasmacytoid dendritic cell neoplasms and so on. The diagnosis and differential diagnosis of myeloid sarcoma are dependent on the pathological and immunohisto-chemical features. The chemotherapy and allogeneic hematopoietic stem cell transplantation of acute myeloid leukemia are the main methods for treatment of myeloid sarcoma.

[Influence of dipyridamole on expression of PAC-1 and CD62p in patients with malignant lymphoma].

This study was purposed to explore the expressions of platelet-activated markers PAC-1 and CD62p in peripheral blood of malignant lymphoma patients and the influence of dipyridamole on their expression. 32 lymphoma patients were divided into simple chemotherapy group (simple group) and chemotherapy plus dipyridamole group (combined group) randomly, and 15 healthy peoples were selected as control group. The dipyridamole of 100 mg/day was given to the patients in combined group. The expression levels of PAC-1, CD62p and fibrinogen (Fib) were detected by flow cytometry and magnetic bead method on day 0, 3, 7 and 14 of chemotherapy respectively. The results showed that the levels of PAC-1, CD62p and Fib in lymphoma patients were significantly higher than those in control group (p < 0.01, 0.05), moreover there was positive correlation between levels of PAC-1 and Fib (r = 0.549, p < 0.01). PAC-1 expression on day 0 and 3 of chemotherapy in simple group was higher than that on day 14 (p < 0.05, 0.01) and CD62p expression on day 3 of chemotherapy was higher than that on day 0, 7 and 14 (p < 0.05, 0.01). PAC-1 expression in combined group on day 14 of chemotherapy was lower than than on day 0 and 3 (p < 0.05, 0.01), and CD62p on day 14 was lower than that on day 3 of chemotherapy (p < 0.05); PAC-1 and CD62p expressions in combined group on day 3, 7 and 14 of chemotherapy were decreased than those in simple group, but Fib level was not changed significantly. It is concluded that the patients with malignant lymphoma usually accompany with platelet activation and hyperfibrinogenemia in peripheral blood. Applying dipyridamole routine dosage in chemotherapy can efficiently restrain platelet activation.

[Relationship between coagulation function and international prognostic index in lymphoma patients].

The study was aimed to investigate the clinical significance of coagulation function changes in lymphoma patients and to analyze the relationship between their changes and international prognostic index (IPI). The prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and fibrinogen (FIB) were detected by magnetic bead method in 75 lymphoma patients and 20 healthy persons. The dehydrogenase (LDH) level was detected by rate method in all lymphoma patients and healthy persons. The results showed that (1) the APTT and FIB more obviously increased in lymphoma patients which displayed as hyperfibrinogenemia, as compared with control group (p < 0.05, p < 0.01); no obvious changes of coagulation indexes presented in patients with different ages and extranodal lesions (p > 0.05, p < 0.01). (2) APTT and FIB levels in stage III and IV patients were much higher than those in the stage II (p < 0.05 and < 0.01), and FIB level in stage IV group was significantly higher than those in the stage III (p < 0.05). FIB level in symptomatic group was significantly higher than that in asymptomatic group (p < 0.01). (3) APTT and FIB in increased LDH group were obviously higher than those in control group (p < 0.05, p < 0.01). Furthermore, FIB in increased LDH group was higher than that in normal LDH group (p < 0.05). FIB in performance status (PS) 2 - 4 groups increased significantly as compared with those in PS 0-1 group (p < 0.01). (4)FIB levels in the low-middle-risk, high-middle-risk and high-risk groups were significantly higher than those in control group (p < 0.01), while FIB levels in high-middle-risk and high-risk groups were higher than those in low-risk group (p < 0.05). (5) the number of FIB increased patients in symptomatic group, increased LDH group, PS 2 - 4 group and Ann Arbor stage III-IV group were much higher than those in counterparts (p < 0.05 or 0.01).There were positive correlations between FIB and LDH level, PS grades, Ann Arbor stages as well as risk grades respectively (p < 0.05 or 0.01). It is concluded that lymphoma patients usually accompany with hyperfibrinogenemia which may be influenced by Ann Arbor stage, systemic symptom, LDH level and PS grade. FIB is supposed to be an effective indication of prognosis in lymphoma patients.