Expression of Ikaros and FUT4 in Children's Acute Lymphoblastic Leukemia and Their Relationship / 中国实验血液学杂志

OBJECTIVE@#To explore the possible molecular mechanism of Ikaros regulation on FUT4 expression by analyzing the correlation of the functional state of Ikaros with level of FUT4 expression, so as to provide the theoretical basis for personalized treatment in children with ALL.@*METHODS@#The subtypes of Ikaros were identified by nested PCR and sequencing. The expression level of FUT4 was detected by quantitative PCR and analyzed by ΔΔCt method in the early stage of treatment, remission and relapse of ALL.@*RESULTS@#Ik1 and Ik2 were the main functional subtypes, and the dominant negative Ikaros was Ik6; the Ik6 was detected in 23 patients with ALL. It was found that 2.73% patients expressing Ik6 alone and 18.18% patients with heterozygous expression were detected. The expression of FUT4 in the newly diagnosed ALL was higher than that in the control group, and the functional Ikaros negatively correlated with the FUT4 expression(r=-0.6329).@*CONCLUSION@#Dominant negative Ikaros closely correlated with the relapse of acute lymphoblastic leukemia in children. The functional Ikaros negatively correlated with FUT4 expression. Ikaros inhibit the transcriptional activity of FUT4, that may be the molecular mechanism of Ikaros regulating the expression of FUT4.

Stage-specific embryonic antigen: determining expression in canine glioblastoma, melanoma, and mammary cancer cells

The expression of stage-specific embryonic antigens (SSEAs) was determined in several types of canine cancer cells. Flow cytometry showed SSEA-1 expression in glioblastoma, melanoma, and mammary cancer cells, although none expressed SSEA-3 or SSEA-4. Expression of SSEA-1 was not detected in lymphoma, osteosarcoma, or hemangiosarcoma cell lines. Relatively stable SSEA-1 expression was observed between 24 and 72 h of culture. After 8 days in culture, sorted SSEA-1⁻ and SSEA-1⁺ cells re-established SSEA-1 expression to levels comparable to those observed in unsorted cells. Our results document, for the first time, the expression of SSEA-1 in several canine cancer cell lines.

Stemness Signature of Equine Marrow-derived Mesenchymal Stem Cells

BACKGROUND: Application of competent cells such as mesenchymal stem cells (MSCs) for treatment of musculoskeletal disorders in equine athletes is increasingly needed. Moreover, similarities of horse and human in size, load and types of joint injuries, make horse as a good model for MSCs therapy studies. This study was designed to isolate and characterize stemness signature of equine bone marrow-derived mesenchymal stem cells (BM-MSCs). METHODS: BM of three mares was aspirated and the mononuclear cells (MNCs) were isolated using density gradient. The primary MNCs were cultured and analyzed after tree passages (P3) for growth characteristics, differentiation potentials, and the expression of genes including CD29, CD34, CD44, CD90, CD105, MHC-I, MHC-II and pluripotency related genes (Nanog, Oct-4, Sox-2, SSEA-1, -3, -4) using RT-PCR or immunocytochemistry techniques. RESULTS: The isolated cells in P3 were adherent and fibroblast-like in shape with doubling times of 78.15 h. Their clonogenic capacity was 8.67±4% and they were able to differentiate to osteogenic, adipogenic and chondrogenic lineages. Cells showed expression of CD29, CD44, CD90, MHC-I and Sox-2 while no expression for CD34, MHC-II, CD105, and pluripotency stemness markers was detected. CONCLUSIONS: In conclusion, data showed that isolated cells have the basic and minimal criteria for MSCs, however, expressing only one pluripotency gene (sox-2).

A Novel Marker for Screening Paroxysmal Nocturnal Hemoglobinuria Using Routine Complete Blood Count and Cell Population Data

BACKGROUND: Final diagnosis of paroxysmal nocturnal hemoglobinuria (PNH) may take years demanding a quick diagnosis measure. We used the facts that PNH cells are damaged in acid, and reagents for measuring reticulocytes in Coulter DxH800 (Beckman Coulter, USA) are weakly acidic and hypotonic, to create a new PNH screening marker. METHODS: We analyzed 979 complete blood counts (CBC) data from 963 patients including 57 data from 44 PNH patients. Standard criteria for PNH assay for population selection were followed: flow cytometry for CD55 and CD59 on red blood cells (RBCs) to a detection level of 1%; and fluorescent aerolysin, CD24 and CD15 in granulocytes to 0.1%. Twenty-four PNH minor clone-positive samples (minor-PNH+) were taken, in which the clone population was <5% of RBCs and/or granulocytes. Excluding PNH and minor-PNH+ patients, the population was divided into anemia, malignancy, infection, and normal groups. Parameters exhibiting a distinct demarcation between PNH and non-PNH groups were identified, and each parameter cutoff value was sought that includes the maximum [minimum] number of PNH [non-PNH] patients. RESULTS: Cutoff values for 5 selected CBC parameters (MRV, RDWR, MSCV, MN-AL2-NRET, and IRF) were determined. Positive rates were: PNH (86.0%), minor-PNH+ (33.3%), others (5.0%), anemia (13.4%), malignancy (5.3%), infection (3.7%), normal (0.0%); within anemia group, aplastic anemia (40.0%), immune hemolytic anemia (11.1%), iron deficiency anemia (1.6%). Sensitivity (86.0%), specificity (95.0%), PPV (52.1%), and NPV (99.1%) were achieved in PNH screening. CONCLUSION: A new PNH screening marker is proposed with 95% specificity and 86% sensitivity. The flag identifies PNH patients, reducing time to final diagnosis by flow cytometry.

Grey zone lymphoma with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin lymphoma: clinicopathologic characterization of 16 cases showing different patterns / 中华病理学杂志

<p><b>OBJECTIVE</b>To profile the clinicopathologic features of a series of grey zone lymphoma (GZL) cases with hybrid features of diffuse large B-cell lymphoma (DLBCL) and classical Hodgkin lymphoma (CHL), with a purpose to gain an in-depth understanding of the borderline B-cell neoplasm.</p><p><b>METHODS</b>The clinical, morphologic and immunophenotyical characteristics of 16 cases were retrospectively analyzed.</p><p><b>RESULTS</b>The patients were mostly male adults, with a male to female ratio of 1.7: 1.0 and a mean age of 40.2 years. Eight patients presented with peripheral nodal lesions and five cases with mediastinal involvement. Histologically and immunophenotypically, the 16 cases were classified into three sub-categories. In 4 cases, the morphologic features resembled CHL more closely, but the neoplastic cells showed uniform and intense positive staining of CD20 (pattern 1). Although the initial impression of the other 8 cases was that of DLBCL, the expression levels of CD20 and PAX5 were variable, and CD30 or CD15 was positive (pattern 2). A characteristic feature of pattern 3, observed in the remaining 4 cases, demonstrated a broad spectrum of morphology with hybrid features of both CHL and DLBCL. The neoplastic cells in pattern 3 were positive for CD20, CD30 and CD15. EBV-LMP1 was detected in 6 of the 11 tested cases. Clinically, most patients with GZL seemed insensitive to immuno-chemotherapy of the R-CHOP regimen.</p><p><b>CONCLUSIONS</b>The diagnostic criteria for GZL with features intermediate between DLBCL and CHL is proposed by the three histologic patterns commonly seen in these lesions. Cases presented with peripheral lesions might differ from those with mediastinal presentation pathologically. At current time, there is no effective treatment for these borderline B-cell lymphomas and the prognosis is poor.</p>

El sistema lewis hoy. nuevos enfoques / The lewis system today. new approaches

La expresión de antígenos (Ags) Lewis depende de alelos heredados en loci independientes, el gen Secretor (SE) que codifica la fucosiltransferasa 2 (FUT2) y el gen Lewis (LE) que codifica la fucosiltransferasa 3 (FUT3). El gen Se codifica una glicosiltransferasa que adiciona una fucosa en la cadena precursora de tipo 1 formando el Ag H en secreciones y fluidos. Como los azúcares inmunodominantes del Ag A y B pueden ser agregados a la cadena H de tipo 1, la FUT2 también controla la expresión de Ag A y B en las secreciones. El gen se es un alelo no funcional. El gen Le codifica una transferasa diferente que adiciona una fucosa en el 2do carbono en el precursor de tipo 1. El alelo le no es funcional. Las FUT2 y FUT3 interactúan para la formación de Ags Lewis en secreciones y fluídos. Los Ags Lewis en los eritrocitos no son en realidad parte integral de la membrana, están adsorbidos sobre la superficie en forma pasiva a partir del plasma. Están ampliamente distribuidos en tejidos humanos, eritrocitos, endotelio, riñón, tracto genitourinario, epitelio gastrointestinal y son receptores para algunos patógenos. Los anticuerpos (Acs) anti-Lewis en general no son clínicamente significativos, aunque se han publicado algunos casos de reacciones transfusionales hemolíticas, enfermedad hemolítica fetoneonatal y rechazo de transplante renal. Este trabajo es una revisión sobre los Ags del Sistema Lewis enfocada hacia sus diferentes funciones biológicas y su importancia en campos variados fuera del Banco de Sangre y la Inmunohematología tradicional.

The expression of Lewis blood group antigens depends on the alleles inherited at independent loci, FUT2 Secretor gene (SE) and FUT3 Lewis gene (LE). The Se and Le alleles encode separate fucosyltransferases that interact to form Lewis antigens in secretions and fluids. The Lewis antigens on red blood cells are not integral to the membrane but are passively adsorbed from the plasma. The allele Se encodes a transferase that adds fucose to type 1 precursor chains in secretions and fluids to form type 1 H antigen. Because A and B terminal sugars may be added to type 1 H chains, FUT2 also controls A and B expression in secretions. The FUT2 allele se gen is a nonfunctional allele. The FUT3 allele Le encodes a transferase that adds a fucose in other position in type 1 H precursor. The FUT3 allele le gen is a nonfunctional allele. The Le antigens are widely distributed in human tissues and fluids and are receptors for some pathogenic organisms. Lewis antibodies are rare and clinically no significant, although there are some reports of hemolytic transfusion reactions, hemolytic disease of the newborn and renal transplant rejection. This review focuses on different biological functions of Lewis antigens and their importance in some fields other than Blood Banks and traditional.

The Molecular Nature of Very Small Embryonic-Like Stem Cells in Adult Tissues

Pluripotent stem cells (PSCs) have been considered as the most important cells in regenerative medicine as they are able to differentiate into all types of cells in the human body. PSCs have been established from several sources of embryo tissue or by reprogramming of terminally differentiated adult tissue by transduction of so-called Yamanaka factors (Oct4, Sox2, Klf4, and cMyc). Interestingly, accumulating evidence has demonstrated the residence of PSCs in adult tissue and with the ability to differentiate into multiple types of tissue-committed stem cells (TCSCs). We also recently demonstrated that a population of pluripotent Oct4(+) SSEA-1(+)Sca-1(+)Lin-CD45(-) very small embryonic-like stem cells (VSELs) resides in the adult murine bone marrow (BM) and in other murine tissue. These very small (~3-6 microm) cells express pluripotent markers such as Oct4, Nanog, and SSEA-1. VSELs could be specified into several tissue-residing TCSCs in response to tissue/organ injury, and thus suggesting that these cells have a physiological role in the rejuvenation of a pool of TCSCs under steady-state conditions. In this review article, we discuss the molecular nature of the rare population of VSELs which have a crucial role in regulating the pluripotency, proliferation, differentiation, and aging of these cells.

The Expression of CD10 and CD15 Is Progressively Increased during Colorectal Cancer Development

BACKGROUND: The aim of this study was to examine the expression of CD10 and CD15 in tumor cells, stromal cells and infiltrating inflammatory cells during colorectal carcinoma (CRC) development and to investigate their expression levels between the tumor center and invasive front and compare them to clinicopathological parameters in invasive CRC. METHODS: We performed immunohistochemical staining for CD10, CD15, and E-cadherin in 42 cases of CRC, 49 of tubular adenoma, 15 of hyperplastic polyp, and 17 of non-neoplastic colon. RESULTS: CD10 was expressed in tumor cells (tCD10), stromal cells (sCD10) and infiltrating inflammatory cells (iCD10), and CD15 was expressed in tumor cells (tCD15) and infiltrating inflammatory cells (iCD15). Their expressions were progressively increased during CRC development and the iCD10 expression level was significantly correlated with the iCD15 expression level in invasive CRC. Invasive front revealed a higher expression level of iCD10 and iCD15 than the tumor center. Moreover, the iCD15 expression level of invasive front was significantly correlated with the degree of tumor budding and tCD15 in whole tissue sections was closely associated with tumor depth. CONCLUSIONS: The present study suggests that the expression of CD10 and CD15 is associated with the development and progression of CRC.

Clinicopathologic study of 40 cases of mediastinal tumours of haematopoietic and lymphoid tissues / 中华病理学杂志

<p><b>OBJECTIVE</b>To study clinical and histopathological features, and diagnosis of mediastinal tumours of haematopoietic and lymphoid tissues (MTHL).</p><p><b>METHODS</b>Forty cases of MTHL were analyzed for clinicopathology by microscopy and immunohistochemical staining and in situ hybridization, according to the updated 2008 WHO classification of tumours of haematopoietic and lymphoid tissues.</p><p><b>RESULTS</b>In 40 cases of MTHL, there were 20 males and 20 females. The ratio of male/female was 1:1. The mean age was 31.8 years and median age was 29 years (range, 12 - 70 years).Superior vena cava syndrome was observed in 28 cases. The specimens of 4 cases were obtained by lumpectomy, whereas 36 cases by biopsy (25 cases by thoracoscopy, 1 by core needle aspiration). Twenty cases lay in anterior mediastinum, and 2 in posterior, 1 in superior, 8 in anterior and superior, 2 in posterior and superior, 2 in anterior and middle, 1 in middle and anterior mediastinum.Frozen section were performed in 28 cases, and 17 cases were diagnosed as tumours of haematopoietic and lymphoid tissues (consistency ratio was 60.7%). Twelve cases were classical Hodgkin lymphomas (cHL) (8 were nodular sclerosis subtype, and 3 were mixed cellarity, 1 was lymphocyte-rich subtype), and 10 were primary mediastinal (thymic) large B cell lymphoma (PMBCL), 10 were precursor lymphocyte neoplasm [8 were T lymphoblastic leukemia/lymphomas (T-LBL), 2 were B-LBL], 1 was MALT lymphoma, 1 was composite lymphoma (PMBCL and cHL), 2 were myeloid sarcomas, 4 were gray zone lymphomas (GZL) (3 had morphology reminiscent of cHL, and 1 of DLBCL, all cases were positive for CD20, PAX5, CD30 and CD15).EBER were detected in 11 cases by in situ hybridization, 2 of which were positive (18.2%), and the 2 positive cases were cHL.</p><p><b>CONCLUSIONS</b>MTHLs occur predominantly in adolescents and young adults, mainly present as superior vena cava syndrome and anterior mediasinal masses. cHL, PMBCL, T-LBL were the most common MTHLs.GZLs mainly occur in young adults, those whose morphology reminiscent of cHL, immunohistochemistry reminiscent of PMBCL, and vice versa. Thoracoscopy, frozen section and a suitable panel of antibodies were practical approaches to MTHL.</p>

[Evaluation of immunoreactivity of cellular markers of CD15 and CD30 among patients with classic Hodgkin's lymphoma]

Hodgkin's lymphoma [HL] is a curable lymphoid malignancy. Different Immunohistochemical markers specially CD15 and CD30 are used to diagnose HL and differentiate it from other lymphomas including anaplastic large cell lymphoma [ALCL]; this study was carried out in patients with Hodgkin's lymphoma to determine the frequency of these markers in Iranian patients with HL. A cross sectional study was done out on all patients with definite diagnosis of classic Hodgkin's lymphoma in the selected hospitals. Patients were classified according to WHO classification of the HL type into: mixed cellularity, nodular sclerosis, lymphocyte rich and lymphocyte depletion subtypes. CD15 and CD30 immunophenotype were detected by monoclonal immunostaining method. The frequency was determined in each group and C.I. [confidence interval] was calculated for the Iranian population. The studies were done on 65 patients. Mean age was 31.9 +/- 18.1 years. 37 [56.9%] were male. CD15 and CD30 was positive in 50 [76.9%] and 58 [89.2%] respectively. Both markers were positive in 46 [70.8%] and were negative in 3 [4.6%]. There was no significant relationship between CD15 and CD30 positivity and age/ gender [p<0.6]. It seems that there is a high frequency of positivity for CD15 or CD30 in Iranian patients with Hodgkin's lymphoma. These markers are useful in diagnosis of Hodgkin's lymphoma. However, in cases who are CD15 negative and CD30 positive, it is better to use additional markers for avoiding misdiagnosis as anaplastic large cell lymphoma

CD20 positivity in classical Hodgkin's lymphoma: Diagnostic challenge or targeting opportunity.

BACKGROUND: It is now well established that Hodgkin cells are clonal B cells with a CD30 and CD15 phenotype. However, on immunohistochemistry, in our experience and the experience of others, CD20 positivity in an otherwise typical classical Hodgkin's Lymphoma is not uncommon and if associated with CD15 negativity poses a potential diagnostic trap and is likely to be called B-NHL. OBJECTIVE: To assess the frequency of B-cell related antigens CD20 and CD79a in classical Hodgkin's Lymphoma. MATERIALS AND METHODS: A total of 91 consecutive cases of classical Hodgkin's Lymphoma were analyzed for co-expression of CD20 and CD79a. Both males and females of all ages were included in this study. All cases of nodular lymphocyte predominant Hodgkin's Lymphoma were excluded. All the cases were stained with a panel of antibodies including LCA, CD20, CD79a, CD30, CD15, CD3, EMA and Alk. Protein. RESULTS: All 91 cases of classical Hodgkin's Lymphoma showed negativity for LCA and positivity for CD30. Eighteen cases (19.8%) showed distinct membrane staining with CD20 in most of the large atypical cells. However, out of these, only 7 cases (7.7%) showed CD79a co-expression, which was largely focal. CD15 negativity with CD20 positivity was seen in 7 (7.7%) cases of otherwise typical classical Hodgkin's Lymphoma. CONCLUSIONS/RECOMMENDATIONS: CD20 expression is frequent in classical Hodgkin's Lymphoma and our results are in consensus with reported literature on this subject. In these cases, LCA negativity of large cells was extremely useful in clinching the right diagnosis.

Molecular confirmation of enterovirus type 71 infection: a post-mortem study of two cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the diagnostic application of molecular detection of enterovirus type 71 (EV71) infection using post-mortem paraffin-embedded tissue.</p><p><b>METHODS</b>Two autopsy cases of EV71 infection were studied by histopathological and immunohistochemical methods. Reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the viral RNA in paraffin-embedded tissue samples.</p><p><b>RESULTS</b>Characteristic features of acute encephalitis were seen in the brain, with most prominent lesions found in the brain stem in both cases. Inflammatory cells were largely CD68-positive microglia with a few CD15-positive neutrophils in the areas of neuronal necrosis. The 5'-untranslated region of EV71 was detected in the medulla by RT-PCR using paraffin-embedded tissues of both cases. Sequencing analysis of the RT-PCR products showed 100% homology to the EV71 strain, recently submitted to the GenBank database from Fuyang, Anhui province.</p><p><b>CONCLUSIONS</b>Molecular detection of EV71 can be performed on formalin-fixed, paraffin-embedded tissue samples from fatally infected patients. Timely and accurate diagnosis of the infection by such molecular approach is crucial for the proper clinical and public health intervention.</p>

Clinically unsuspected Hodgkin's lymphoma diagnosed primarily from bone marrow trephine biopsy: report of six cases.

Bone marrow may be the initial or rarely the only site of involvement in Hodgkin's lymphoma. A high index of suspicion is required to pick up the histopathological changes of Hodgkin's lesions in the bone marrow like necrosis, presence of Reed-Sternberg cell or its variant in a polymorphic background infiltrate, focal fibrosis and myxoid change especially in the absence of classical clinical picture. Bone marrow with immunohistochemistry has a valuable role in the staging and in the diagnosis of primary medullary Hodgkin's lymphoma. B-symptoms may easily masquerade as an infectious process as in all our cases the patients had fever as a presenting feature, in four of them tuberculosis was suspected clinically and two had received antitubercular therapy elsewhere. We report six human immunodeficiency virus-negative patients diagnosed over a period of 5 years in which the initial diagnosis of Hodgkin's lymphoma was suggested from bone marrow histology.

Extramedullary infiltration of acute monocytic leukemia/monoblastic sarcoma: a clinicopathologic and immunophenotype analysis of 5 cases / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features, diagnosis and differential diagnosis of extramedullary infiltration of acute monocytic leukemia/monoblastic sarcoma.</p><p><b>METHODS</b>Five cases of extramedullary infiltration of acute monocytic leukemia/monoblastic sarcoma were selected from 102 cases of myeloid sarcoma diagnosed during the period from 1990 to 2006. The clinicopathologic findings and followup data were retrospectively analyzed. Immunohistochemical study was also carried out with SP method.</p><p><b>RESULTS</b>Among the 5 cases studied, 3 were males and 2 were females, including 2 children and 3 adults. Generalized lymphadenopathy was found in 4 patients and skin lesions were observed in 2 patients. The tumor cells in all cases were positive for CD68 (KP1), CD68 (PGM1), lysozyme and CD45. They were negative for MPO, CD15, CD163, TdT, CD117, T and B cell markers. The Ki-67 index ranged from 40% to 80%. Follow-up data were available in all the 5 patients. Four of the 5 patients died of the disease, with the average survival time being 6.25 months.</p><p><b>CONCLUSIONS</b>Monoblastic sarcoma is a rare disease with poor prognosis. It is almost impossible to distinguish monoblastic sarcoma from granulocytic sarcoma and other types of small round cell tumors on the basis of morphologic examination alone. Immunohistochemistry is mandatory for a correct diagnosis.</p>

Lymphocyte predominance hodgkin lymphoma clincopathological and immunohistochemical interpretations using CD15 and CD20

Hodgkin disease is a lymphoid tumor that accounts for less than [1%] of all DeNovo neoplasms occurring every year worldwide. The aims of the study was to assess the expression of immunohistochemical markers namely [CD[20], CD[15] in malignant cells of Hodgkin's disease. Over the period extending from [January 2006-March 2008], specimens form 31 cases of lymphocyte predominance Hodgkin lymphoma [Rye's classification] as formalin fixed, paraffin embedded tissue blocks from excisional lymph node biopsies. For each case, five representative sections were prepared. One stained with hematoxyline and eosine [HxE], and four other sections [on positively charged slides] prepared for IHC procedures using CD[15], and CD[20]. For the evaluation of markers expression, a semiquantitative evaluation system was used to register the staining intensity and the numbers of positive cells. The most common site of lymph node biopsy was the cervical lymph node in [61.2%] of cases. Immunohistochemically, CD[20] was expressed in [12.9%] of cases, while CD[15] was expressed in [77.4%] of cases. This study has shown that HD with a nodular growth pattern and a lymphocyte-rich background encompasses 2 entities with distinct morphologic, phenotypical features. Therefore, the precise classification of such case requires a combination of conventional histology and immunohistology with distinct panel of antibodies

Immunophenotypic and cytogenetic features in 51 cases of chronic lymphocytic leukemia / 中国实验血液学杂志

The study was aimed to investigate the immunophenotypic and cytogenetic features of chronic lymphocytic leukemia (CLL) in order to provide an evidence for diagnosis and therapy. Immunophenotypic analysis was performed by using a panel of monoclonal antibodies and three-color immunofluorescence staining methods of flow cytometry in 51 patients with CLL, and the cytogenetic features were analyzed by R-banding technique. The results indicated that among 51 CLL cases, the positive rate of CD19 and CD23 was 96.1%, followed by CD15 (94.1%), CD20 (82.4%) and CD22 (78.4%). The positive rate of CD38 was 23.5%. Forty-six patients expressed both CD5 and CD19. Seven main clonal chromosomal abnormalities were detected by conventional cytogenetics (CC) in eighteen cases (35.3%), with three cases of +12, two cases of 13q(-), other chromosomal abnormalities included +14, 6q(-), t (11; 14), t (14; 18) and t (2; 7). Expression of the antigens had no relationship with chromosomal abnormalities. It is concluded that typical CLL express CD5, CD19 and CD23, and the positive rate detected by CC in CLL is low. Immunophenotyping in combination with cytogenetic technique plays an important role in the diagnosis and prognosis of CLL.

diagnostic value of monoclonal antibody panel in cases of Hodgkin's lymphoma

The interface between Hodgkin's lymphoma [HL] and B-cell NHL has become more ambiguous in recent years. Yet, the clinical presentation, prognosis, and treatment requirement of Hodgkin's lymphoma are very different from most B-cell NHL. Thus, distinguishing between those lymphomas is still mandatory. Reviewing and immunophenotyping [using a monoclonal antibody panel] of cases diagnosed by routine histologic sections stained with hematoxylin and eosin [H and E] as Hodgkin's lymphoma, subtyping the cases in view of the new classification based on morphologic and immunophenotypic data, as well as studying the efficacy of human fascin antibody as a new marker for Reed-Sternberg cells. All cases were submitted to pathological examination of the routine H and E stained section as well as immunohistochemical evaluation of fascin, CD30, CD 15, CD20, and CD3 expression with subsequent re-evaluation of cases and establishing a final diagnosis based on morphologic and immunophenotypic data. In the present work, positive immunostaining for fascin was expressed in 97.7% of classic HL, CD30 in 80.9%, CD 15 in 70.2%, and CD20 in 10.6%. Only 26 cases [57.7%] showed coexpression of CD30 and CD15. After the review of immunohistochemical slides the diagnosis was confirmed in 37 cases of classic HL. Problematic cases were reclassified into CHL of mixed cellularity subtype [2 cases], CHL of lymphocyte rich subtype [2 cases], and CHL of the lymphocyte depletion subtype [2 cases]. Two additional cases were classified as T-cell rich large B-cell lymphoma [TCRBCL]. Two cases remained unclassified after immuno staining. The present study emphasized that immunohistochemistry supports and refines the H and E based diagnosis of Hodgkin's lymphoma and that Fascin is a sensitive marker for RS cells and may be used to differentiate between Hodgkin's and non-Hodgkin's lymphoma in difficult cases. Furthermore, complete immunohistochemical panel is needed to distinguish between T-cell rich B-cell lymphoma and lymphocyte rich-classic Hodgkin's lymphoma whether nodular or diffuse taking into consideration that absence of CD 30 or CD 15 immuno staining or expression of CD20 does not 'rule out the diagnosis of classic Hodgkin 's lymphoma and that coexpression of CD 30 and CD 15 when coupled with CDS expression necessitates complementary studies

Interfollicular Hodgkin's disease: an uncommon pattern that may cause diagnostic difficulty.

Interfollicular Hodgkin's Disease is characterised by reactive follicular hyperplasia with involvement of the interfollicular area of lymph node by Hodgkin's lymphoma. It represents a peculiar pattern of focal involvement of lymph node and does not constitute a classical subtype. Its importance rests in the fact that it can be misinterpreted as one of the many causes of reactive hyperplasia of lymph node and not as Hodgkin's disease. Eleven cases of interfollicular Hodgkin's disease were diagnosed in a period of five years. Majority of the patients were less than twenty years and all had localised lymphadenopathy. Lymph node biopsy showed follicular hyperplasia with expanded interfollicular area. Careful search of the interfollicular area showed infiltration by inflammatory cells and scattered Reed-Sternberg and Hodgkin's cells. Immunohistochemistry with CD 15 and CD 30 highlighted the atypical cells. This report emphasises on the problems in diagnosis of interfollicular Hodgkin's disease.

Cell phenotypes, immunoglobulins and complement in lesions of eumycetoma caused by madurella mycetomatis

Mycetoma [maduromycosis] is a common health problem in Sudan. The causative organisms are either true fungi [eumycetoma] or actinomycetes [actinomycetoma]. The commonest eumycetoma in Sudan is caused by M mycetomatis. The cell phenotypes, immunoglobulins and complement in lesions of M mycetomatis were characterized by immunohistochemistry. In the HandE sections there were three types of inflammatory reactions. Type I reaction consisted of three zones: a neutrophil zone surrounding the grain, an intermediate zone of macrophages and giant cells and a peripheral zone consisting of lymphocytes and plasma cells. The neutrophils stained positively for CD15. The macrophages were positive for CD68. The majority of cells in the outermost zone were CD3 positive [T lymphocytes]; they were rimmed by CD20 positive cells [B lymphocytes]. In type II reaction there was no neutrophil zone, the grain being surrounded by macrophages and giant cells that stained positive for CD68. Type III reaction consisted of a discrete epithelioid granuloma without wellformed grains. IgG, IgM and C3 were found on the surface of the grain and the hyphae