抗凝血相关因子抗体检测方法的建立及临床应用

目的抗磷脂抗体综合征(APS)患者血清中是否存在识别纤溶酶、凝血酶和活性蛋白C(APC)的抗磷脂抗体(aPL)。方法分别用凝血酶、纤溶酶和APC包被酶标板,建立抗纤溶酶、抗凝血酶和抗APC抗体的检测方法。结果在40例APS患者和40名正常对照的血清中,前者抗纤溶酶抗体阳性率为42.5%(17/40),抗凝血酶抗体阳性率为35%(14/40),对照组则两者均为阴性,差异有显著性(P<0.01)。但两组样本抗APC抗体检测结果的A值经检验差异无显著性(P>0.05)。结论APS患者的血清中可检测出抗纤溶酶、抗凝血酶和抗APC抗体,其临床意义有待进一步研究。     

SYMPTOMATIC COLITIS AS THE INITIAL PRESENTATION OF WEGENER'S GRANULOMATOSIS

SUMMARY Wegener's granulomatosis is a multisystem disorder which rarely presents with gastrointestinal involvement. We report a case in which colitis was the reason for hospital admission. Diagnosis may be difficult unless clinical, serological and histopathological features are all considered.     

Rapid detection of Flt3 mutations in acute myeloid leukemia patients by denaturing HPLC

BACKGROUND: fms-related tyrosine kinase 3 (Flt3) is the most commonly mutated gene in human acute myeloid leukemia (AML) and has been implicated in its pathogenesis. Because screening of Flt3 in AML patients by PCR followed by gel electrophoresis is time-consuming and fails to detect some very small internal tandem duplications (ITDs), we developed a method for screening of FLT3 receptor mutations with PCR plus denaturing HPLC (D-HPLC). METHODS: Total mRNAs extracted from 34 AML patients were first analyzed for the presence of juxtamembrane length mutations and tyrosine kinase domain point mutations by a conventional method involving PCR amplification, restriction enzyme digestion, and agarose gel electrophoresis (PCR-RED-AGE). Subsequently, the same patient panel was analyzed by D-HPLC, using specifically designed primers and optimized running temperatures for the length and point mutation analysis. RESULTS: Thirty-four patients were analyzed by PCR-RED-AGE; 9 were positive for known Flt3 mutations: 6 of 34 (18%) for ITDs in exon 14 and 3 of 34 (9%) for point mutations in exon 20. The same patient panel was analyzed by D-HPLC, and additional nucleotide changes were discovered; in total, 14 sequence variations were identified: 7 of 34 (21%) for ITDs in exon 14; 2 of 34 (6%) for point mutations in exon 20; 1 of 34 (3%) for a new point mutation in exon 16; and 4 of 34 (12%) for polymorphisms in exons 13 and 14. Direct sequencing analysis identified nucleotide alterations in each of the "D-HPLC positives" but in none of the "D-HPLC negatives", yielding a specificity and sensitivity of 100% for D-HPLC-based screening. CONCLUSIONS: This novel D-HPLC-based procedure, which is optimized for identification of new point mutations in the catalytic and regulatory domains of FLT3 receptor, could potentially be useful for studies involving precise genotype determination, which could be critical for selection of innovative AML therapies targeting the FLT3 protein.     

Primary cutaneous lymphomas: a reprisal

Primary cutaneous lymphomas (PCLs) are a group of lymphoid neoplasms provided with heterogeneous clinical, histological, immunohistochemical and molecular features. They can be classified in two groups: cutaneous T-cell lymphomas (CTCLs) and cutaneous B-cell lymphomas (CBCLs). Recent studies show an increase of the incidence of PCLs over the last three decades. Our aim is to evaluate the commonest types of PCL analysing the clinical characteristics, histology, phenotype, molecular biology, prognosis and therapy.     

Lymphoma classification: the quiet after the storm

The classification of malignant lymphomas remained controversial for over 30 years. The first scheme was proposed by Rappaport in the '60th and was based on incorrect histogenetic concepts. To overcome these limitations, several groups formulated new proposals in '70th. Among these two merited attention: the Lukes and Collins and the Kiel Classifications. They were based on the assumption that each lymphoma category might be related to a precise differentiation step of the lymphoid system, thus excluding any correlation with histiocytes, present on the Rappaport scheme. The Kiel Classification became very popular in Europe, while the one of Luke and Collins did not meet success in the United States (U.S.). In 1978, the National Cancer Institute proposed an international trial to compare the classifications used in Europe and U.S. The result was the genesis of the Working formulation, the tool for lymphoma classification in the U.S. up to the early '90th, but which was conversely rejected in Europe. In order to get over this lack of transatlantic communication, in 1994 the Revised European-American Lymphoma (REAL) Classification was proposed by the International Lymphoma Study Group. Its goal was to list "real" entities, each defined by the presence of homogeneous morphologic, phenotypic, cytogenetic, molecular, and clinical criteria, along with the possible recognition of its normal counterpart. The REAL Classification became the model for the WHO Classification of all haematopoietic tumours published in 2001. The present review aims to analyse future perspectives after the fourth edition of the WHO Classification released in 2008.     

Partial nodal involvement by marginal zone lymphoma. Use of IGK gene rearrangement analysis in diagnostic work-up

Nodal marginal zone lymphoma (NMZL) is an indolent B-cell lymphoma that originates from the marginal zone of B-cell follicles. The tumour is rather uncommon, and shares some morphologic and immunophenotypic similarities with the extranodal form of marginal zone lymphomas. However, diagnosis of NMZL implies the exclusion of lymphoplasmacytic lymphoma, follicular lymphoma, and lymph node involvement by extra nodal or splenic marginal zone B-cell lymphoma In addition, its distinction from reactive conditions, including T-zone hyperplasia, are sometimes problematic based on morphologic grounds. We describe a patient who presented with cervical and inguinal lymphadenopathies and high inflammation indexes. Bone marrow and lymph node biopsies were performed for definitive diagnosis. Bone marrow histological and immunophenotypic examinations were normal and excluded haematological disease. In contrast, lymph node evaluation showed some features compatible with a possible lymphoproliferative disorder, even though no definite diagnosis could be made based on morphologic and immunohistochemical investigation. In particular, the problem of a differential diagnosis between NMZL and a florid hyperplasia of monocytoid B-elements was posed. Thus, in order to assess the nature (neoplastic vs. reactive) of the lesion, molecular analysis of the immunoglobulin genes was performed by PCR. Notably, although no clonal rearrangements were revealed by IGHV@ analysis, further evaluation of the immunoglobulin light chain (IGKV@) confirmed the presence of a clonal B-cell population. Accordingly, a final diagnosis of NMZL was made. In conclusion, this case is a good example of the crucial role of complete molecular analysis in the diagnostic work up of lymphoproliferative disorders.     

Dental Expression and Role in Palliative Treatment

World Health Organization defines palliative care as the active total care of patients whose disease is not responding to curative treatment. Palliative care for the terminally ill is based on a multidimensional approach to provide whole-person comfort care while maintaining optimal function; dental care plays an important role in this multidisciplinary approach. The aim of the present study is to review significance of dentist''s role to determine whether mouth care was effectively assessed and implemented in the palliative care setting. The oral problems experienced by the hospice head and neck patient clearly affect the quality of his or her remaining life. Dentist plays an essential role in palliative care by the maintenance of oral hygiene; dental examination may identify and cure opportunistic infections and dental disease like caries, periodontal disease, oral mucosal problems or prosthetic requirement. Oral care may reduce not only the microbial load of the mouth but the risk for pain and oral infection as well. This multidisciplinary approach to palliative care, including a dentist, may reduce the oral debilities that influence the patient''s ability to speak, eat or swallow. This review highlighted that without effective assessment of the mouth, the appropriate implementation of care will not be delivered. Palliative dental care has been fundamental in management of patients with active, progressive, far-advanced disease in which the oral cavity has been compromised either by the disease directly or by its treatment; the focus of care is quality of life.     

Immunohistochemical and other prognostic factors in B cell non Hodgkin lymphoma patients, Kampala, Uganda

Background  

Non Hodgkin lymphomas are the most common lymphomas in Uganda. Recent studies from developed countries have shown differences in survival for the different immunophenotypes. Such studies are lacking in Africa where diagnosis is largely dependent on morphology alone. We report immunohistochemical and other prognostic factors in B cell non Hodgkin lymphoma patients in Kampala, Uganda.     

The bone marrow stroma in hematological neoplasms--a guilty bystander

In the setting of hematological neoplasms, changes in the bone marrow (BM) stroma might arise from pressure exerted by the neoplastic clone in shaping a supportive microenvironment, or from chronic perturbation of the BM homeostasis. Under such conditions, alterations in the composition of the BM stroma can be profound, and could emerge as relevant prognostic factors. In this Review, we delineate the multifaceted contribution of the BM stroma to the pathobiology of several hematological neoplasms, and discuss the impact of stromal modifications on the natural course of these diseases. Specifically, we highlight the involvement of BM stromal components in lymphoid and myeloid malignancies, and present the most relevant processes responsible for remodeling the BM stroma. The role of bystander BM stromal elements in the setting of hematological neoplasms is discussed, strengthening the rationale for treatment strategies that target the BM stroma.