OBJECTIVETo investigate the role of cytogenetic analysis in the detection of bone marrow (BM) involvement in patients with non-Hodgkin's lymphoma (NHL).

METHODSThe bone marrow samples of 74 patients with NHL were detection by using morphology, cytogenetic test, flow cytometry and molecular biological assay. The detected results of morphology, cytogenetic test, flow cytometry and molecular biological assay alone and thier combined detection were compared, the detective rate and consistencies of the 4 methods were analyzed.

RESULTSThe detection rates of BM involvement by using morphology, cytogenetic, flow cytometry, and molecular biological assays were 21.6%, 17.6%, 23.0% and 33.8% respectively. The detective rate was enhanced to 44.6% by combining the 4 methods. Cytogenetic test showed the result consistent with the other methods.

CONCLUSIONAlthough cytogenetic test shows a lower detective rate than the other methods, but in some patients the cytogenetic test can detect the abnormality of bone marrow which can not be detected by other methods alone, the combination test of 4 detection methods can enhance the detectable rate of BM involvement.

Bone Marrow ; pathology ; Bone Marrow Examination ; Cytogenetic Analysis ; Flow Cytometry ; Humans ; Lymphoma, Non-Hodgkin ; diagnosis ; genetics

The aim of this study was to evaluate the significance of id4 gene promotor methylation detection in NHL patients. MS-PCR method was used to detect the status of id4 gene methylation in health donors and newly diagnosed NHL patients. The results indicated that the id4 gene was unmethylated in bone marrow samples from health donors. Among 18 newly diagnosed NHL patients, including one NHL patient with bone marrow cells involved, 4 patients were found in id4 gene methylation by MS-PCR. The 14 patients with id4 gene unmethylation were in their stable status and no bone marrow involvement were found by bone marrow biopsy during the 8-month follow-up. During the follow-up, the patient with both bone marrow involvement and id4 gene methylation turned to leukemia, in 2 out of the 3 patients with id4 gene methylation but without bone marrow involvement at diagnosis, the bone marrow involvement was found at last. It is concluded that the id4 gene methylation may be an indicator for MRD in NHL patients without bone marrow involvement.
Bone Marrow ; pathology ; DNA Methylation ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; Lymphoma, Non-Hodgkin ; genetics ; pathology ; Promoter Regions, Genetic ; genetics

Paraffin-embedded tissue samples from 30 cases of non-Hodgkin's lymphoma(NHL) and 10 of reactive hyperplasia, were processed for interphase cytogenetic chromosomal study. We performed non-fluorescent in situ hybridization(NFISH) using the enzymatic method with digoxigenin-labeled DNA centromeric probes for chromosome 7,12,18 and X, and a painting probe for chromosome 18. Chromosomal aberrations were observed in 27(90%) out of 30 cases of NHL. The most commonly observed numerical aberration was extracopy of X chromosome. There were some characteristic aberrations corresponding to each grade and group of NHL by International Working Formulation: In low grade NHL(9 cases), a third were associated with extracopy of chromosome 12, and disomy X was frequently found in small lymphocytic lymphoma(75%). With intermediate grade(16 cases), tetraploidy(25%), translocation of chromosome 18(25%), and extracopy of chromosome 18(19%) were characteristically associated. These results suggest that interphase NFISH is an easily performable method in retrograde cytogenetic study of archival materials. Some specifically correlated chromosomal aberrations corresponding to the histopathologic grades and groups could provide us more valuable information for determining pathologic diagnosis and assessing the clinical outcome of NHL.
*Chromosome Aberrations ; Human ; Immunophenotyping ; *In Situ Hybridization ; Interphase ; Lymphoma, Non-Hodgkin/*genetics/pathology ; Paraffin Embedding ; Pseudolymphoma/genetics/pathology

To determine whether the p53 expression might be a predictor for treatment sponse and overall survival in nodal non-Hodgkin's lymphoma (NHL), we analyzed e expression of p53 in 69 NHL patients. p53 protein expression was analyzed by munohistochemistry with long-term follow up (1-148 months: median 12.2). p53 pression was noted in 23/69 (33.3%) patients. Complete response (CR) rate to stemic chemotherapy was correlated with stage (I/II) (p=0.038), but not with 3 expression (p=0.2856). Poor overall survival was associated with stage =0.0010) or IPI score (p=0.0076), but not with p53 expression (p=0.8601). From ratification analysis by stage, in stage III/IV patients, the p53 positive oup had a trend to be associated with poor overall survival than the p53 gative group. Multivariate analysis revealed that p53 positive group was sociated with less CR rate compared to the p53 negative group (p=0.046), ereas overall survival was correlated with stage (p=0.0320), not with p53 atus. p53 expression was associated with less CR rate in patients with DLBL. rther studies with large numbers of samples and homogenous group of NHL are eded to determine the prognostic value of cell cycle regulator, p53 in NHL.
Antibodies, Monoclonal ; Cell Cycle Proteins/biosynthesis ; Female ; Gene Expression ; Human ; Immunohistochemistry ; Immunophenotyping ; Lymph Nodes/pathology* ; Lymph Nodes/metabolism* ; Lymphoma, Non-Hodgkin/pathology* ; Lymphoma, Non-Hodgkin/metabolism* ; Lymphoma, Non-Hodgkin/genetics ; Lymphoma, Non-Hodgkin/drug therapy ; Male ; Middle Age/Mpartment of Microbiology ; Prognosis ; Protein p53/immunology ; Protein p53/genetics ; Protein p53/biosynthesis*

The study was aimed to investigate the BCL-XL expression and mutation, and its clinical significance in non-Hodgkin's lymphoma. Lymphoma cells were selectively isolated by laser microdissection. BCL-XL expression from lymphoma tissue and microdissected lymphoma cells was measured by using real-time quantitative reverse transcription-polymerase chain reaction. BCL-XL mutation was analyzed by using direct sequencing of PCR products. The results showed that compared to 15 patients with reactive hyperplasia, BCL-XL was overexpressed in follicular lymphoma (n = 30), both in lymphoma tissue (P = 0.0064) and in microdissected lymphoma cells (P < 0.0001). No significant rise of BCL-XL expression was observed in patients with T-cell lymphoma (n = 24) and diffuse large B cell lymphoma (n = 24). In follicular lymphoma, high BCL-XL level was associated with multiple extranodal involvement (P = 0.0004), elevated lactate dehydrogenase level (P = 0.0019), high-risk international prognostic index (P = 0.0013) and a short overall survival time (P = 0.0451). Mutation analysis revealed one synonymous mutation (Codon 109 ACA-->ACC) in one case of follicular lymphoma patient. It is concluded that BCL-XL expression is closely correlated with progress of follicular lymphoma and prognosis of patients with follicular lymphoma. The value of BCL-XL expression as a prognostic marker in follicular lymphoma should be considered.
Base Sequence ; Humans ; Lymphoma, Follicular ; genetics ; pathology ; Lymphoma, Non-Hodgkin ; genetics ; pathology ; Molecular Sequence Data ; Point Mutation ; bcl-X Protein ; biosynthesis ; genetics

OBJECTIVETo investigate the relationship between histopathological subtype of non-Hodgkin' s lymphoma(NHL) and chromosomal abnormalities, and compare the difference of chromosomal abnormalities between China and the West.

METHODSRoutine G banding chromosome analysis and fluorescent in situ hybridization( FISH) were performed on lymph node specimens from 155 NHL patients.

RESULTSDiffuse large B-cell lymphoma( DLBCL) constituted 38.1% of the cases followed by follicular lymphoma(FL) 17.4% , small lymphocytic lymphoma( SLL) 10.3% , peripheral T-cell lymphoma ( PTCL) ( unspecified) 8.4%, and angioimmunoblastic lymphoma 7.1%. One hundred nineteen patients (76. 8%) had clonal chromosomal abnormalities. The incidence of chromosomal abnormalities among FL, SLL, DLBCL, anaplastic large cell lymphoma (ALCL) and precursor T-cell lymphoblastic lymphoma (TLBL) was 96.3% , 87.5% , 86.4%, 83.3% and 83.3%, respectively. Complex karyotype was 86. 3% in DLBCL. The most frequent structural abnormalities in DLBCL involved chromosomes 3, 6, 14 and 1, with had high frequencies of 3q27 (41.2%) and 6q21, 6q23, 6q25 involvement (23.5%). But only 2 cases of DLBCL had classical t( 14; 18) (q32;q21) which was lower than that in the West (20%). The positive rate of IgH rearrangement was 40. 1% in DLBCL by FISH. No 13q14 deletion was found in 16 cases of SLL. Normal karyotype was observed in 8/11 cases with angioimmunoblastic T-cell lymphoma patients.

CONCLUSIONSThe distribution of the histopathological subtypes of NHL is different among different geographical areas. The chromosomal abnormalities in DLBCL was comparable between China and the West, but t( 14; 18) was infrequent in the former. The chromosomal abnormalities in SLL was different from the West. The incidence of chromosomal abnormalities in angioimmunoblastic T-cell lymphoma was lower than that in the West.

Adult ; Chromosome Structures ; Cytogenetic Analysis ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; classification ; genetics ; pathology ; Lymphoma, Non-Hodgkin ; classification ; genetics ; pathology ; Male

OBJECTIVETo analyze and optimize the gene rearrangement primers of different frame regions (FR) of immunoglobulin heavy chain (IgH) genes by bioinformatic methods and explore the application of these primers in the detection of paraffin-embedded lymphoma tissues.

METHODSThree pairs of primers from IgH FR1, FR2 and FR3 regions (P1c, P2A and P31, respectively) were selected as the B cell gene rearrangement primers after comparison of the gene fragments in 44 IgH variable and 6 joining regions. Using one pair of T cell receptor (TCR) gamma primer as the T cell gene rearrangement primer, 101 histopathologically confirmed lymphoproliferative samples including 80 B cell lymphomas, 14 T cell lymphomas, and 7 reactive proliferative lymph nodes were examined by PCR for gene arrangement. The DNAs from DG75 and Jurkat cell lines were used as the positive controls for B and T cell lymphoma, respectively, with those from reactive proliferative lymph nodes as the negative control.

RESULTSThe positivity rates of IgH primers (P1c, P2A and P31) in the 80 B cell lymphomas were 37.5% (30/80), 52.5% (42/80) and 70.0% (56/80), respectively, and only one of the 14 T cell lymphoma cases was positive for the primers, suggesting significant differences in the detection rates of B cell lymphomas by the 3 primers. The detection rate was increased to 83.9% by combining the results by P31 and P2A primers. No positivity was found in the proliferative reaction tissues.

CONCLUSIONPrimers from IgH FR3 region genes are more sensitive than that from the FR1 and FR2 regions in the detection of gene rearrangement in paraffin-embedded lymphoma tissues. The detection rates can be increased by combining the results with the primers for IgH FR3 with that of FR2.

DNA Primers ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; genetics ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Lymphoma, B-Cell ; diagnosis ; genetics ; pathology ; Lymphoma, Non-Hodgkin ; diagnosis ; genetics ; pathology ; Lymphoma, T-Cell ; diagnosis ; genetics ; pathology ; Male ; Paraffin Embedding

OBJECTIVETo investigate the expression, localization and interrelationship of P27(kip1) and cyclin D3 in non-Hodgkin's lymphoma (NHL).

METHODSThe expressions of P27(kip1), cyclin D3 and index Ki-67 was detected in 100 NHL and 20 reactive lymph nodes by immunohistochemical technique. The expression and localization of P27(kip1) and cyclin D3 in 3 NHL cell lines were detected by Western blot, double immunolabelling and laser scanning confocal microscopy, respectively.

RESULTSIn general the expression of P27(kip1) in NHL was lower than in control group, and was negatively related to the tumor aggressiveness and proliferating activity; the expression of cyclin D3 in NHL was higher than in control group, and was positively related to the tumor aggressiveness and proliferating activity. There was a negative correlation between P27(kip1) and cyclin D3. Nevertheless, anomalous high P27(kip1) expression was found in a few NHL tissues with high expression of cyclin D3 and Ki-67. Overexpression and colocalization of P27(kip1) and cyclin D3 was found in Raji cell line.

CONCLUSIONSUnder expression of P27(kip1) and overexpression of cyclin D3 may play a role in the occurrence and development of NHL. Anomalous high P27(kip1) expression and its interaction with cyclin D3 may be another mechanism for tumor genesis of NHL.

Cell Line, Tumor ; Cyclin D3 ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; metabolism ; Cyclins ; genetics ; metabolism ; Humans ; Lymphoma, Non-Hodgkin ; genetics ; metabolism ; pathology

The metastasis-suppressing role of the nm23 gene in the metastatic spread of malignant tumor is still debated. We examined the nm23-H1 protein expression and gene mutation in non-Hodgkin's lymphomas to compare with the clinicopathologic parameters. The expression of nm23-H1 protein was immunohistochemically examined in 150 cases of non-Hodgkin's lymphomas; 85 diffuse large B cell lymphomas (DL-BCL), 18 marginal zone B cell lymphomas (MZL), 3 mantle cell lymphomas, 25 peripheral T cell lymphomas, not otherwise specified (TCLNOS), and 19 NK/T cell lymphomas (NK/T). Eighty-one cases (58 DLBCL, 6 MZL, 4 TCLNOS, and 13 NK/T) were studied for nm23-H1 gene mutation in exon 1 to 5. The high expression of nm23-H1 protein was associated with the high IPI score (p=0.019) and the low survival rate of the patients (p=0.0039). The gene mutation of nm23-H1 was detected in 10.3% of DLBCL and 30.7% of NK/T; but none in MZL and TCLNOS. The mutation was found in exon 1 in 5 cases, exon 2 in two cases, exon 4 in one case and both exon 1 and 2 in two cases. Our results suggest that the expression of nm23-H1 protein can be used as a poor prognostic marker in non-Hodgkin's lymphomas, and the mutational change of gene may operate in the lymphomagenesis.
Tissue Array Analysis ; Survival Analysis ; Prognosis ; Polymorphism, Single-Stranded Conformational ; Nucleoside-Diphosphate Kinase/*genetics/metabolism ; Mutation/*genetics ; Middle Aged ; Male ; Lymphoma, T-Cell/genetics/metabolism/pathology ; Lymphoma, Non-Hodgkin/genetics/metabolism/*pathology ; Lymphoma, Mantle-Cell/genetics/metabolism/pathology ; Lymphoma, Large-Cell, Diffuse/genetics/metabolism/pathology ; Lymphoma, B-Cell/genetics/metabolism/pathology ; Immunohistochemistry ; Humans ; Female ; DNA Mutational Analysis ; Base Sequence

OBJECTIVETo study the cytogenetic characteristics of B cell non-Hodgkin's lymphoma (B-NHL) with bone marrow involvement, and to explore the clinical significance and prognosis.

METHODSClinical data of 126 B-NHL patients with bone marrow involvement diagnosed in our hospital were retrospectively analyzed. Chromosome banding analysis was performed after 24 h culture.

RESULTS(1) The B-NHLs included were diffuse large B-cell lymphoma (DLBCL) 38.9% (49 cases), lymphoplasmacytic lymphoma (LPL) 19% (24 cases), mantle cell lymphoma (MCL) 16.7% (21 cases), follicular lymphoma (FL) 9.5% (12 cases), marginal zone lymphoma (MZL) 8.7% (11 cases) and small lymphocytic lymphoma (SLL) 7.1%(9 cases). (2) Chromosome aberrations (CA) were detected in 52 of 126 patients (41.3%) by conventional cytogenetics (CC), including clonal CA 38 cases, and non-clonal CA 14 cases. Ploidy levels in 38 clonal CA cases were pseudodiploid (57.9%), hypodiploid (15.8%) and hyperdiploid (26.3%). The incidence of chromosomal abnormalities among DLBCL, MCL, MZL, LPL, FL and SLL was 73.4%, 38.1%, 36.4%, 8.3%, 8.3% and 11.1%, respectively. (3) Clonal CA, CA more than two kinds, and CA of chromosomes 2, 3, 9, 11, 17, 18 and 20 were associated with shorter overall survival (OS) in DLBCL. More than two kinds of CA and CA of chromosome 3, 13 were associated with shorter OS in MCL.

CONCLUSIONSThe incidence of CA was higher in aggressive lymphoma than in indolent lymphoma. Complex CA were quite common, and some specific CA might have prognostic significance.

Adolescent ; Adult ; Aged ; Bone Marrow ; pathology ; Child ; Child, Preschool ; Chromosome Aberrations ; Female ; Humans ; Lymphoma, Non-Hodgkin ; classification ; genetics ; pathology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Young Adult