OBJECTIVETo detect the clonal relationship, the rearrangement, and the mutational status of IgVH gene; the influence of these molecular characteristics on the clinical outcome in Hodgkin variant of Richter syndrome; and the possible molecular pathogenesis in this transformation.

METHODSThe clonal rearrangements and mutational status of IgVH genes were analyzed in Hodgkin variant of Richter syndrome and B-CLL with Reed-Stemberg (R-S)-like cells by GeneScan analysis and sequencing. Semi-nest PCR based on laser capture microdissection was utilized to compare the clonal relationship between B-CLL and R-S/R-Slike cells. Immunohistochemical staining was used to detect the different expressions of ZAP70, p53, IRF-4 and LMP1 in the two components.

RESULTS(1) 5/6 B-CLL cases transformed to Hodgkin lymphoma (HL)/R-S-like cells carried the mutated IgVH genes; (2) 2 cases of R-S cells and 1 case of R-S-like cells were clonally distinct from B-CLL clone and express LMP1, whereas 1 case of R-S-like cells was relating to the surrounding B-CLL cells and did not express LMP1; (3) 2/6 B-CLL cases transformed to HL convey VH4-34 and VH3-48 respectively.

CONCLUSIONS(1) Richter transformation to HL/R-S-like cells evolves from the B-CLL which originates from the germinal center or post germinal center B cells, indicating that different lymphoma cells of different subtypes in Richter syndrome come from different B cell lineage and possibly involve a different pathogenesis and pathway; (2) HL and R-S-like cells evolve from either the B-CLL clone or may develop as a clonally unrelated lymphoma, the independent secondary malignancies are appear to be EBV-positive, possibly as a consequence of the underlying immunodeficiency; (3) The biased usage of IgVH genes suggested a role of antigens involved in the HL variant of Richter syndrome.

Aged ; Aged, 80 and over ; Clone Cells ; pathology ; Female ; Herpesvirus 4, Human ; Hodgkin Disease ; classification ; genetics ; pathology ; virology ; Humans ; Immunoglobulin Variable Region ; genetics ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; pathology ; Male ; Middle Aged ; Mutation ; Reed-Sternberg Cells ; pathology ; Syndrome

OBJECTIVETo study the clonal rearrangements and mutation status of IgVH genes in classic Richter's syndrome, the relationship between molecular findings of IgVH gene and clinical outcome, and to deciper the possible molecular mechanism of transformation.

METHODSThe clonal rearrangements and mutation status of IgVH genes were analyzed in cases of classic Richter's syndrome by Genescan and sequencing. Immunohistochemical study for zeta-chain associated protein kinase 70 kDa (ZAP70), p53 and interferon regulation factor 4 (IRF-4) was also performed.

RESULTSSamples of 18 cases of B-chronic lymphocytic leukemia (B-CLL)/ diffuse large B-cell lymphoma (DLBCL,78. 3%) had identical tumor cell clones, whereas DLBCL developed as a clonally independent neoplasm in 5 patients (21.7%). Among the clonally related group, 12 cases carried unmutated VH genes in both B-CLL and DLBCL components and VH3-23, VH3-74 and VH1-2 were accounted for the B-CLL transformation to DLBCL. Immunohistochemical study showed that the transformed DLBCL expressed CD5 in 32.1% of cases, CD23 in 14.3%, ZAP70 in 23.8%, p53 in 80.6% and IRF-4 in 82.6% of the cases respectively. Follow-up data were available in 17 patients with classic Richter's syndrome. The median survival period was 7 months. No significant difference in survival rate was obtained between the clonally related or unrelated groups, between IgVH gene mutated or unmutated groups, and between the groups with or without expression of ZAP70, p53 and IRF-4.

CONCLUSIONSThe ratio of clonally related transformed DLBCL from B-CLL to clonally unrelated DLBCL is 2:1. Clonal transformation to DLBCL predominantly occurs in B-CLL patients carrying unmutated IgVH genes. The biased IgVH gene usage suggests antigens are involved in classic Richter's syndrome. Molecular differences of IgVH genes and very poor clinical outcome of this group of transformed DLBCL indicate that there cases may be regarded as a distinct subset of DLBCL.

Aged ; Aged, 80 and over ; B-Lymphocytes ; pathology ; Genes, p53 ; genetics ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; Lymphoma, B-Cell ; genetics ; Lymphoma, Large B-Cell, Diffuse ; genetics ; Male ; Middle Aged ; Somatic Hypermutation, Immunoglobulin ; genetics ; ZAP-70 Protein-Tyrosine Kinase ; genetics

OBJECTIVETo investigate the rate of dual rearrangements of lymphocytic antigen receptor genes in non-Hodgkin lymphomas (NHL) and its pathogenesis and pathologic significance.

METHODSPCR analysis of monoclonal, polyclonal and dual rearrangements of IgH and TCR gamma, TCR beta genes was carried out in 125 cases of NHL to evaluate the rate of dual rearrangements, immunohistochemistry was performed for a Ki67 protein expression in 117 cases and the proliferation index was calculated. The relationship between antigen receptor gene rearrangements and proliferation index was analyzed.

RESULTSCombination of the two pairs of IgH gene primers with the multiplex PCR for TCR gamma and TCR beta gene revealed dual rearrangements in 8% (8/96) of B-NHL, 17% (5/29) of T-NHL. In B cell NHL, IgH gene monoclonal, dural and polyclonal rearrangements were identified in 65, 8 and 15 cases respectively, while in T-cell NHL, they were in 15, 5 and 9 cases, respectively. There was no significant difference between proliferation index and monoclonal, dual, polyclonal rearrangements in both B-NHL and T-NHL by One-way test.

CONCLUSIONDual rearrangements in NHL are not rare and have no relationship with proliferation index.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carrier Proteins ; genetics ; Cell Proliferation ; Child ; Female ; Gene Rearrangement ; Genes, T-Cell Receptor gamma ; genetics ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; metabolism ; Lymphoma, B-Cell ; genetics ; metabolism ; pathology ; Lymphoma, Non-Hodgkin ; genetics ; metabolism ; pathology ; Lymphoma, T-Cell ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Polymerase Chain Reaction ; Receptors, Antigen ; genetics ; Young Adult

To search for bioactive compounds from the flower disc of Helianthus annuus L., chromatography was used to isolate and purify the chemical constituents, their structures were identified by spectral analysis, MTT method was applied to investigate their cytotoxic activities, some compounds showed moderate cytotoxic activities on SF-268, MCF-7 and HepG2 cell lines. Eleven compounds were obtained from the flower disc of H. annuus, and identified as ent-kaurane-2alpha, 16alpha-diol (1) and entkaurane-15alpha,16alpha-epoxy-17-al-19-oic acid (2), and nine known diterpenes, ent-kaurane-16P-ol (3), phyllocladan-16beta-ol (4) , ent-atisan-16a-ol (5) , grandifloric acid (6) , angeloylgrandifloric acid (7), ent-kaurane-16-en-19-oic acid (8), ent-kaurane-17-hydroxy-15-en-19-oic acid (9), ent-kaurane-16beta, 17-dihydroxy-19-oic acid (10), and ciliaric acid (11). Compounds 1 and 2 are new compounds, some compounds showed cytotoxic activities on SF-268, MCF-7 and HepG2 cell lines.
Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Crystallization ; Diterpenes ; chemistry ; isolation & purification ; pharmacology ; Diterpenes, Kaurane ; chemistry ; isolation & purification ; Flowers ; chemistry ; Helianthus ; chemistry ; Humans ; Inhibitory Concentration 50 ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry ; Transition Temperature