OBJECTIVETo analyze the clinicopathological features and prognostic factors of patients with diffuse large B-cell lymphoma(DLBCL).

METHODSNinety-four cases of DLBCL followed up were selected in Fujian Tumor Hospital. The immunohistochemistry method was used to detect the protein expressions of BCL-2 BCL-6, MYC, CD10 and MUM-1, the gene abnormalities of MYC and BCL-2 were analyzed by fluorescence in situ hybridization, and the clinical pathological features and the related factors affecting prognosis in the patients with DLBCL were analyzed.

RESULTSThe protein positive rates of BCL-2, BCL-6, MYC, CD10 and MUM-1 in 94 patients were 75.53% (71/94), 58.51% (55/94), 52.13% (49/94), 15.96% (15/94) and 34.04% (32/94) respectively. The detection rate of MYC gene abnormality was 20.93% (9/43) and the detection rate of BCL-2 gene abnormality was 44% (22/50); 2 kinds of gene abnormalities were of multiple copies, and 2 cases (2.13%) were abnormal in MYC and BCL-2 genes simultaneously. The median survival time of 3 years in 94 patients was 21.79 months (2-36 months), and the overall survival rates of 1 and 3 years were 82.98% and 64.89% respectively. Single factor analysis revealed that the high ECOG score (≥ 2), high international prognostic index (IPI) classification, positive expression of BCL-6 protein, and MYC and BCL-2 gene simultaneously abnormal were the risk factors influencing the prognosis (all P<0.05). COX regression analysis showed that IPI classification, ECOG score and treatment methods were independent factors influencing the prognosis (all P<0.05).

CONCLUSIONIPI classification, ECOG score and treatment methods have greater impacts on the prognosis of patients with DLBCL. Chemotherapy combined with radiotherapy or surgical treatment can significantly improve the prognosis of patients.

Genes, myc ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; Proto-Oncogene Proteins c-bcl-6

The antidiabetic drug metformin has been found to have beneficial effects in various neurological disorders; however, the molecular mechanisms underlying these effects remain unclear. Here we report that metformin protects neuronal cells from quinolinic acid (QUIN)-induced excitotoxicity. For this, we pretreated N18D3 neuronal cells with metformin prior to QUIN for 24 h. We found that pretreating the cells with metformin significantly improved cell survival rate in a concentration-dependent manner and reduced apoptotic cell death, as revealed by a MTT assay and DAPI staining, respectively. Calcium imaging using fluo-4 showed that metformin (100 µM) inhibited the intracellular calcium increase that was induced by QUIN. In addition, mRNA expression of pro-apoptotic genes, p21 and Bax, was decreased and of anti-apoptotic genes, Bcl-2 and Bcl-xl, was increased with metformin treatment compared to QUIN-induced cells. The immunoreactivity of phosphorylated ERK1/2 was elevated in cells treated with metformin, indicating the ERK1/2 signaling pathway in the neuroprotective effects of metformin in QUIN-induced cell death. Collectively, our data demonstrates that metformin exerts its neuroprotective effects by inhibiting intracellular calcium increases, allowing it to regulate ERK1/2 signaling and modulate cell survival and death genes.
Apoptosis ; Calcium ; Cell Death ; Cell Survival ; Genes, bcl-2 ; Metformin ; Nervous System Diseases ; Neurons ; Neuroprotection ; Neuroprotective Agents ; Quinolinic Acid ; RNA, Messenger

The present study evaluated the effects of Androctonus amoreuxi scorpion venom, Cerastes cerastes snake venom and their mixture on prostate cancer cells (PC3). An MTT assay was used to determine the anti-proliferative effect of the venoms, while quantitative real time PCR was used to evaluate the expression of apoptosis-related genes (Bax and Bcl-2). Furthermore, colorimetric assays were used to measure the levels of malondialdehyde (MDA) and antioxidant enzymes. Our results show that the venoms significantly reduced PC3 cell viability in a dose-dependent manner. On the other hand, these venoms significantly decreased Bcl-2 gene expression. Additionally, C. cerastes venom significantly reduced Bax gene expression, while A. amoreuxi venom and a mixture of A. amoreuxi & C. cerastes venoms did not alter Bax expression. Consequently, these venoms significantly increased the Bax/Bcl-2 ratio and the oxidative stress biomarker MDA. Furthermore, these venoms also increased the activity levels of the antioxidant enzymes, catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase. Overall, the venoms have cytotoxic and anti-proliferative effects on PC3 cells.
Apoptosis ; Catalase ; Cell Survival ; Gene Expression ; Genes, bcl-2 ; Glutathione Peroxidase ; Glutathione Reductase ; Hand ; Humans* ; Malondialdehyde ; Oxidative Stress ; Prostate* ; Prostatic Neoplasms* ; Real-Time Polymerase Chain Reaction ; Scorpion Venoms ; Scorpions* ; Snake Venoms* ; Snakes* ; Superoxide Dismutase ; Venoms ; Viper Venoms ; Viperidae*

OBJECTIVETo investigate the effect of BCL-2/MYC double-hit on prognosis in diffuse large B-cell lymphoma（DLBCL）.

METHODSA retrospective study was conducted to investigate clinical and pathological data of 111 patients with DLBCL. CD10, BCL-6, MUM-1, BCL-2 protein expressions were examined by immune-histochemical methods, and abnormal BCL-2 and MYC genes were analyzed by FISH for patients with sufficient pathological data. SAS 8.2 was adopted to perform Chi- square test, COX's proportional Hazard Model, Life table survival analyses.

RESULTSOf 111 patients, male 77 cases, female 34 cases, the median age was 55（14-85）years, CD10, BCL-6, MUM-1, BCL-2 positive rates were 15.7%（16/102）, 58.8%（60/102）, 33.0%（34/103）, 74.8（77/103）respectively, the abnormal rate of BCL-2 gene was 43.1%（25/58, 24 cases with multiple copies, 1 case with translocation）, and the abnormal rate of MYC gene was 20.4%（10/49, 10 cases with multiple copies）. Coexistence of BCL-2 and MYC genes abnormalities accounted for 13.0%（6/46）. According to the classification of Hans model, GCB subgroup accounted for 41.2%（42/102）, and non-GCB subgroup 58.8%（60/102）, the median survival time was 24 months, 3-year and 5-year overall survival rates were 48.5% and 39.7% respectively. Overall survival rates of normal and abnormal BCL-2 gene were 34.2%，22.8%, respectively with no statistical significance（P=0.770）. Overall survival rates of normal and abnormal MYC gene were 35.9% and 22.2% ，with no statistical significance（P=0.650）. Overall survival rate of double-hit was 0, far worse than that of single abnormal gene（P=0.034）, which implied double-hit of BCL-2 and MYC gene abnormality to be adverse prognostic factors. BCL-6 protein express could be classified as benign prognostic factors, while ECOG score≥2, escalated IPI index as adverse prognostic factors, and further COX risk model regression analysis indicated that ECOG score, IPI grading and treatment methods were independently adverse factors affecting prognosis. Comprehensive therapy based on chemotherapy could improve outcome.

CONCLUSIONBCL-2/MYC genes double-hit was the factor for the adverse outcome in DLBCL patients. However, ECOG score, IPI risk grading and treatment methods were the independent factors affecting prognosis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Genes, myc ; Humans ; Lymphoma, Large B-Cell, Diffuse ; Male ; Middle Aged ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; Retrospective Studies ; Survival Rate ; Translocation, Genetic ; Young Adult

OBJECTIVE: To investigate the process of apoptosis in lungs and liver induced by crushing hindlimbs of rat, and study the mechanism of crush injury. METHODS: The rat experimental model of hindlimbs crush injury was established. The cell apoptosis in lungs and liver was detected by TUNEL assay, and the expression of Bax, Bcl-2 and caspase-3 apoptin was examined by immunohistochemistry. RESULTS: Compared with the control group, the partial muscle injury of rat's hindlimbs was more serious with more apoptosis observed in lungs and liver (P < 0.05). The expression of Bax was up-regulated and Bcl-2 was down-regulated, whereas caspase-3 expression was activated (P < 0.05). CONCLUSION The cell apoptosis has increased significantly in lungs and liver after crush injury of hindlimbs in rat. The correlation factor released during tissue injury may mediate apoptosis process.
Animals ; Apoptosis/physiology* ; Caspase 3/metabolism* ; Genes, bcl-2 ; Hindlimb/injuries* ; Immunohistochemistry ; Liver/physiopathology* ; Lung/physiopathology* ; Rats ; Up-Regulation ; bcl-2-Associated X Protein

OBJECTIVETo explore the expression of Bcl-2 mRNA and its effect on prognosis of patients with primary gastrointestinal diffuse large B-cell lymphoma (PGI-DLBCL).

METHODSReal time quantitative PCR was used to determine the expression of Bcl-2 mRNA in 40 PGI-DLBCL patients and 17 healthy controls. The association of Bcl-2 expression with clinicopathological features and prognosis of the patients was analyzed.

RESULTSThe expression level of Bcl-2 mRNA in PGI-DLBCL patients was 1.03 ± 0.93, significantly higher than that of the controls (0.41 ± 0.21) (P < 0.05). The expression of Bcl-2 mRNA in stage IIE-IV patients (1.28 ± 1.01) was significantly higher than that in the stage I-II2 patients (0.62 ± 0.61) (P < 0.05). The expression of Bcl-2 mRNA in patients with international prognostic index (IPI) score >2 (1.95 ± 1.27) was significantly higher than those with IPI score ≤ 2 (0.86 ± 0.75)(P < 0.05). The expression of Bcl-2 mRNA in patients with complete remission (CR) (0.71 ± 0.58) was significantly lower vs. 2.42 ± 0.91 in patients with no CR (P < 0.05). Univariate analysis indicated that β2-MG, IPI score>2, the Lugano staging, and Bcl-2 mRNA expression were associated with overall survival (OS) and progression-free survival (PFS) (P < 0.05). Multivariate analysis indicated that IPI score>2 was independently associated with OS (P < 0.05), and both IPI score >2 and Bcl-2 mRNA expression were independently associated with PFS (P < 0.05).

CONCLUSIONSThe expression of Bcl-2 mRNA in the tumor tissue of PGI-DLBCL patients is significantly higher than that in controls. PGI-DLBCL patients with higher expression of Bcl-2 have a poor chemotherapy response and inferior prognosis. IPI score >2 and higher expression of Bcl-2 mRNA are independent poor prognostic factors for PFS in PGI-DLBCL patients.

Disease-Free Survival ; Genes, bcl-2 ; Humans ; Lymphoma, B-Cell ; diagnosis ; genetics ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; genetics ; metabolism ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism

OBJECTIVETo identify and investigate clinicopathological features of B cell lymphomas with concurrent myc and bcl-2/IgH or bcl-6 translocations ("double-hit" lymphoma).

METHODSTissue microarray was constructed from formalin-fixed and paraffin-embedded tissue samples of aggressive B cell lymphomas diagnosed between 2009 and 2012, including 129 cases of diffuse large B cell lymphoma (DLBCL), 5 cases of B-cell lymphoma, unclassifiable with features intermediate between diffuse large B-cell lymphoma and Burkitt lymphoma (BCLU), 7 cases of Burkitt lymphoma and 4 cases of high-grade follicular lymphoma with diffuse large B cell lymphoma component. Interphase fluorescence in-situ hybridization (FISH) was performed with a panel of probes including myc, bcl-2/IgH and bcl-6 to document related gene translocation and copy number changes. Medical record review was performed and follow-up data was recorded.

RESULTSAmong 145 cases, 5 cases (3.4%) of B cell lymphomas with concurrent myc and bcl-2/IgH or bcl-6 rearrangements (double-hit lymphomas) were identified, including 2 cases involving myc and bcl-2 translocations (1 DLBCL and 1 BCLU), and 3 cases involving myc and bcl-6 translocations (all DLBCLs). Three cases with concurrent bcl-2/IgH and bcl-6 translocations were found. Single gene translocations or increase of copy numbers were found in 66 cases, representing 51.2% (66/129) of all de novo DLBCLs. Ki-67 index of the 5 "double-hit" lymphomas ranged from 60% to 100%. Clinical follow-up data were available in 4 of the 5 "double-hit" lymphoma patients, three of whom died within 2 years and 1 patient was alive after 36 months of follow-up.

CONCLUSIONS"Double-hit" B-cell lymphomas are rare and can only be identified by molecular detection. They should not be considered synonymous with BCLU morphologically, and may present entities within other morphological spectra. Most of the patients have a poor prognosis. Further in-depth studies of larger case numbers are required to determine the pathologic and genetic variables of the lesion.

Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Burkitt Lymphoma ; drug therapy ; genetics ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Genes, bcl-2 ; Genes, myc ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, B-Cell ; drug therapy ; genetics ; Lymphoma, Follicular ; drug therapy ; genetics ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; genetics ; Male ; Middle Aged ; Prednisone ; therapeutic use ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Proto-Oncogene Proteins c-bcl-6 ; genetics ; Proto-Oncogene Proteins c-myc ; genetics ; Retrospective Studies ; Translocation, Genetic ; Vincristine ; therapeutic use

OBJECTIVETo perform immunophenotyping and molecular genetic analysis for diffuse large B-cell lymphoma (DLBCL), and to explore their correlation and implication for prognosis.

METHODSImmunohistochemical streptavidin peroxidase (SP) method was used to determine the expression of CD10, BCL6 and MUM1 in 59 cases of DLBCL. A Hans algorithm was used to classify DLBCL into germinal center B-cell (GCB) and non-GCB subtypes. Interphase fluorescence in situ hybridization (FISH) assay was performed on paraffin-embedded lymphoma tissue sections to detect translocations and amplifications of BCL6, BCL2 and MYC genes with dual-color break-apart BCL6 probe, dual-color dual-fusion IgH/ BCL2 probe and dual-color break-apart MYC probe, respectively.

RESULTSIn the 59 cases of DLBCL, 28.8% (17/59) belonged to GCB subtype, and 71.2% (42/59) belonged to non-GCB subtype. The incidences of BCL6, BCL2 and MYC gene translocations were 24.1% (14/58), 1.7% (1/59) and 5.3% (3/57), respectively. The incidences of BCL6, BCL2 and MYC gene amplifications were 17.2% (10/58), 22.0% (13/59) and 21.1% (12/57), respectively. BCL6 amplification was not correlated with BCL6 translocation (P=0.424), but was correlated with amplifications of BCL2 and MYC (C=0.405 and 0.403, respectively, P <0.01). The incidence of BCL6 translocation in GCB type was higher than that in non-GCB type, and amplifications of BCL6, BCL2 or MYC were more frequently encountered in non-GCB type, though no statistical significance was detected (P=0.089 and 0.106, respectively). By univariate analysis, immunophenotyping and international prognostic index (IPI) exerted a significant effect on overall survival (OS) (P=0.047 and 0.001, respectively), but to which BCL6 translocation and amplification of the 3 genes were not related (P=0.150 and 0.444, respectively). By multivariate analysis, IPI score was the only independent prognostic factor for OS (RR =3.843, P=0.017).

CONCLUSIONThe GCB subtype of DLBCL is less common in the patient cohort. Common genetic aberrations have included BCL6 translocation and BCL6, BCL2 and MYC amplifications. Amplification of the 3 genes is strongly correlated with each other, and the incidence of BCL2 translocation is low. Immunophenotyping only has minor significance for the prognosis. Genetic aberrations cannot predict the clinical outcome of DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA-Binding Proteins ; genetics ; Female ; Genes, bcl-2 ; Genes, myc ; Humans ; Immunophenotyping ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; genetics ; immunology ; Male ; Middle Aged ; Proto-Oncogene Proteins c-bcl-6

OBJECTIVETo study the expression of miR-16 and bcl-2 in T lymphoblastic lymphoma/leukemia (T-LBL/ALL) and its relationship to prognosis.

METHODS70 cases of T-LBL/ALL with follow-up data were studied by using immunohistochemical EnVision method for CD1a, CD3, cCD3, CD7, CD10, CD20, CD23, CD34, CD43, CD45RO, CD99, TDT, MPO, bcl-2 and Ki-67. The expression levels of miR-16 were examined by TaqMan real-time polymerase chain reaction (RT-PCR). Thirty cases of reactive lymph node were selected as control.

RESULTSAmong the 70 cases of T-LBL/ALL, the percentages of tumor cells expression of TDT, CD99, CD3, CD7, CD10, CD34, CD1a, cCD3, bcl-2, CD45RO and CD43 were 94.3% (66/70), 94.3% (66/70), 68.6% (48/70), 92.9% (65/70), 32.9% (23/70), 24.3% (17/70), 40.0% (28/70), 51.4% (36/70), 34.3% (24/70), 37.1% (26/70), and 48.6% (34/70). Separately, while tumor cells expression of MPO, CD20 and CD23 was all negative. A figure of Ki-67 expression > 80% was found in 24 cases and ≤ 80% in 46 cases. The expression of miR-16 was up-regulated in T-LBL/ALL, and it was 5.07 times of the reactive lymph node(P = 0.001). The high expression group of miR-16 was significantly correlated with longer over survival (P = 0.041). The prognosis of negative bcl-2 group was better than bcl-2 positive one(P = 0.904). The relationship of miR-16 and bcl-2 was significant(P = 0.042,χ(2) = 4.147). Survival multivariate COX proportional hazard regression analysis revealed that the low expression of miR-16 might be a independent poor prognosis factor (P = 0.049).

CONCLUSIONSWhile the high expression group of miR-16 has longer OS than that in low expression group. The prognosis of bcl-2 negative was better than bcl-2 positive. miR-16 may be a independent prognosis factor. The relationship of miR-16 and bcl-2 might suggested that gene regulation may be influenced by them.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Genes, bcl-2 ; Humans ; Immunohistochemistry ; Infant ; Male ; MicroRNAs ; metabolism ; Middle Aged ; Neoplasm Staging ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Survival Rate ; Young Adult

Animals ; Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Gene Rearrangement ; Genes, myc ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; genetics ; Prednisone ; therapeutic use ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; genetics ; metabolism ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; Translocation, Genetic ; Vincristine ; therapeutic use