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Article
The role of Wilm̛̛s Tumor1 immunohistochemical marker in surface epithelial ovarian tumors
دور المناعي النسيجي للمعلم WT1 في أورام المبيض الطلائية السرطانية

Authors: Lubab F. Talal لباب فاضل --- Sahira A. Ali ساهرة عبود --- Muna I. AL Hafedh منى اسماعيل
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2015 Volume: 57 Issue: 2 Pages: 145-150
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Wilms’ tumor 1 is a tumor suppressor gene. The gene is located in chromosome 11p13. And its expression was found in many solid tumors (including ovarian tumor) and also expressed in hematologic malignancies, Recent studies found that WT1 to be involved in angiogenesis.Objectives: To evaluate the expression of WT1 in surface epithelial ovarian tumorand study the possibility of using WT1 as replacement of both;ovarian tumor marker CA125 and a endothelial cell phenotypic marker CD34.Patients and methods: This is a study of a retrospective ( cross sectional ) of sixty cases with total abdominal hysterectomy and bilateral salpingo - oopherectomy collected from department of Histopathology – Teaching Laboratories / Medical City Teaching Hospital , as well as Al alwya hospital and Al Habibia hospital in Baghdad during the period of study from December 2007 to December 2012. Thirty cases diagnosed as surface epithelial ovarian tumors and thirty cases of histologically normal ovarian tissue which were included as a control group. Formalin - fixed, paraffin - embedded ovarian tissue blocks from 60 cases were used . Three section of 4 micron for each taken and stained with WT1, CD34, and CA125 immunohistochemical marker on positively charged slides.Results: there were a significant correlation between expression of WT1 and histological types of surface epithelial ovarian tumor with a higher expression in serous tumors among other cancer types (P-value < 0.001).There was a significant positive correlation between the expression of WT1 and CA125 scores ( p-value < 0.001).There was a significant correlation between WT1microvessel density (MVD) expression and CD34- microvessel density (MVD) expression in ovarian tumors (P-value = 0.05).On the other hand, there were no significant correlation of WT1 with the age of cases (P-value = 0.9) and with the grade of ovarian tumors ( P-value = 0.23) . Conclusions: The present study demonstrates high expression of WT1 in both tumor and endothelial cells in surface epithelial ovarian tumors, and it had dual usages in evaluation of both ovarian tumor cells and the vascular density. That was proved by demonstrating a significant correlation between WT1 and CA125 expression, and between WT1-MVD and CD34- MVD . There was no statistically significant association between WT1 expression and different tumor grades. There was significance differences in WT1expression among different histological subtypes of primary ovarian carcinomas, with serous carcinoma as the most frequent type. Key words: Ovarian tumor; WT1; CA125; CD34.

خلفية عن الدراسة: المعلم المناعيWT1 هوعامل نسخ ويقع في الكروموسوم 11p13, اكتشف لأول مرة في الورم الكلوي ((Wilms حيث انه يعمل كمثبط للجينات السرطانية , كما يوجد في عدة أورام من ضمنها أورام المبيض وأورام الدم الخبيثة, وفي الآونة الأخيرة وجد بان WT1 له أيضا دور في تكوين الأوعية الدموية.هدف الدراسة: صممت هذه الدراسة للبحث عن العلاقة بين المعلمات المناعية WT1 ،CA125, CD34 وأورام المبيض الطلائية السطحية ودراسة إمكانية استخدام المعلم المناعيWT1 كبديل لكل من CA125و .CD34 المرضى وطرائق العمل: شملت هذه الدراسة )60( حالة تراجعية لاستئصال الكامل للرحم والمبايض والأنابيب والتي تم جمعها على شكل بلوكات نسيجية مطمورة بالشمع ومثبتة بالفورمالين جيدا , من قسم النسيج المرضي في مختبرات التعليمية /مدينة الطب بالإضافة إلى مستشفى العلوية ومستشفى الحبيبية في بغداد خلال الفترة من ديسمبر 2007إلى ديسمبر2012.تم دراسة ثلاثين حالة من أورام المبيض الطلائية السطحية بالإضافة إلى ثلاثون نسيج مبيض طبيعي خالي الورم والتي استخدمت كمجموعة سيطرة, قد عملت ثلاثة مقاطع نسيجية على شرائح زجاجية ذات شحنة موجبة لكل ٍ من المعلماتWT1، CD34، وCA125النتائج: كان هناك علاقة كبيرة بين المعلم المناعي WT1والنوع النسيجي لأورام المبيض الطلائي السطحي مع تعبير العالي في الأورام النسيجية المصلية بين أنواع السرطان الأخرى قيمة ( p >0.001) . ووجد بان هناك علاقة إيجابية ذات دلالة إحصائية بين المعلم المناعي WT1وCA125p<0.001)). وأيضا هناك علاقة كبيرة بين WT1-MVD و CD34- MVD في أورام المبيض (P = 0.05). أما من الناحية الأخرى فانه لم يكن للمعلم المناعي WT1 أي علاقة مع العمر في الحالات التي تمت دراستها فقيمة p= 0.9) ) , كما انه لا توجد أية علاقة مع درجات تمايز الورم في المبيض . (P = 0.23) الاستنتاجات: توضح هذه الدراسة التعبيرات العالية للمعلم المناعي WT1في كل من الخلايا السرطانية والخلايا البطانية للأوعية الدموية في أورام المبيض الطلائية السطحية ، وكان للمعلم المناعي WT1 استخدام مزدوج في تقييم كل من الخلايا السرطانية في المبيض وكثافة الأوعية الدموية.وقد اثبت ذلك من خلال وجود علاقة موجبة بين المعلمين WT1وCA125 ، وبين كثافة الأوعية الدموية الصغيرة المقاسة بواسطة المعلمين WT1وCD34 لا توجد علاقة بين المعلم المناعي WT1و درجات تمايز ورم المبيض.وقد وجد بان هنالك علاقة كبيرة في تعبير المعلم المناعي WT1 و مختلف الأنواع النسيجية لأورام المبيض مع تعبير عالي ل WT1للنوع النسيج المصلي الخبيث.مفتاح الكلمات:ورم المبيض ؛ WT1؛ CA125؛ CD34.


Article
The Prognostic Role of (CD34) In The Angiogenesis of Endometrial Adenocarcinoma

Author: Lubab Fadhil Talal
Journal: Iraqi Academic Scientific Journal المجلة العراقية للاختصاصات الطبية ISSN: 16088360 Year: 2006 Volume: 5 Issue: 1 Pages: 77-86
Publisher: The Iraqi Borad for Medical Specialization المجلس العراقي للاختصاصات الطبية

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Abstract

ABSTRACT: BACKGROUND: Tumor angiogenesis is essential for tumor growth and metastases. Recently ,studies showed that microvessel density (MVD),a measure of tumor angiogenesis has found to have a prognostic significance for predicting metastases and survival in endometrial adenocarcinoma and identifying patients at high risk for recurrences . The aim of this study is to understand the correlation between the MVD and various clinicopathological factors in endometrial adenocarcinoma in order to evaluate the role of angiogenesis in the assessment of the biological behavior of endometrial adenocarcinoma and to prove that MVD can be used as a valuable prognostic factor and can detect high risk patients for post-operative recurrence who may need adjuvant therapy after surgery . PATIENTS AND METHODS: Forty-one (41) cases were included in this study ; one case is normal endometrium in the proliferative phase , 5 cases with sever endometrial hyperplasia and 35 cases with endometrial adenocarcinoma taken from the Pathology Department at the Medical City and Private Pathology Laboratory between the years 2000-2004 . Paraffin blocks cut in two sections ; one for Haematoxylin & Eosin stain and the other for CD34 immunostaining slides were studied well for tumor grade ,stage ,myometrial invasion and pelvic and paraaortic lymph nodes metastases . The mean MVD was studied by scanning the tumor sections at a low power X40 and X100 then identifying areas of highest neovascularization (hot spots) .Five hot spot areas were selected from each section ,and the MVD were counted at X200 magnification (x10 ocular ,x20 objective) by taking the mean of those microvessels. The correlation between mean MVD and various clinicopathological factors were studied including the age ,tumor grade ,stage ,myometrial invasion and pelvic and paraaortic lymph nodes metastases . RESULTS : CD34 is a reliable and sensitive endothelial cell marker used in the assessment of tumor angiogenesis The study showed that a significant statistical correlation was found between the mean MVD and patients age , tumor grade and myometrial invasion. While there were no statistical correlation significance between the mean MVD and tumor stage and pelvic & paraaortic lymph nodes . CONCLUSION : We conclude that MVD is important in the assessment of the biological behavior of endometrial adenocarcinoma , where it is an indicator for the emergence and growth of tumor and it is important to detect patients at high risk of recurrences and to decide the post-operative adjuvant therapy


Article
Immunohistochemical analysis of CD34 to evaluate angiogenesis in chronic lymphocytic leukemia
التعبير المناعي النسيجي الكيميائي للدالة CD34 لتثمين تكوين الاوعية الدموية الدقيقة في ابيضاض الدم اللمفاوي المزمن

Author: Shaymaa M. Abdelateef شيماء محمد عبد اللطيف
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2013 Volume: 55 Issue: 2 Pages: 131-134
Publisher: Baghdad University جامعة بغداد

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Background: Chronic lymphocytic leukemia (CLL) results from a progressive accumulation of long-lived, functionally incompetent, nonproliferating lymphocytes.Angiogenesis is defined as the formation of new capillaries from pre-existing blood vessels and plays an important role in the progression of solid tumors as well as several hematologic malignancies like CLL.Patients and methods: A retrospective cross-sectional study done on 68 patients with CLL compared with 15 control individuals (anemic patients), all recruited at the Medical City Teaching Laboratories from January 2005 to December 2008. The bone marrow biopsy (BMB) of each was re-examined histologically. Immunohistochemical (IHC) technique was performed on BMB sections utilizing monoclonal mouse CD34 class II antibody. Results: This study revealed higher microvessel density (MVD) in BM of CLL patients than normal control marrows. CLL patients had a mean MVD of 10.78 VS 4.26 miorovessel count/high power field (MVC/HPF) in control subjects. Significant inverse correlation was found between MVD and PCV levels and platelet count. A Significant positive correlation was found between MVD and the percentage of lymphocytes in the bone marrow, BM infiltration pattern and the modified Rai stage of CLL patients Conclusions: Angiogenesis in CLL BM was significantly higher than control marrows and was more accentuated with advanced clinical stage of the disease.Key words: Chronic lymphocytic leukemia; CD34; IHC

الخلفية : ابيضاض الدم اللمفاوي المزمن ينتج من التجمع التراكمي للخلايا اللمفاوية طويلة العمر والتي وظيفيا غير فعالة و غير قابلة للانقسام. تكوين الاوعية الدموية الدقيقة هو تكون الشعيرات الدموية الجديدة من اوعية دموية سابقة و يلعب دوراً مهما في تقدم الاورام الصلبة وكذلك العديد من الاورام الدموية مثل ابيضاض الدم اللمفاوي المزمن.الهدف من الدراسة: تقييم تكوين الاوعية الدموية الدقيقة في ابيضاض الدم اللمفاوي المزمن وذلك بحساب كثافة الاوعية الدموية الدقيقة باستخدام الدالة CD34.اعداد و طرائق العمل: شملت هذه الدراسة التراجعية ثمان وستون حالة من ابيضاض الدم اللمفاوي المزمن (48 ذكر و 20 انثى). كانت لديهم خزع لنقي العظم اجريت اجريت في المختبرات التعليمية في مدينة الطب مابين الفترة من كانون الثاني2005 الى كانون الاول 2008 كذلك تم دراسة مجموعة ضابطة (العدد 15 حالة).تم استخدام التقنية المناعية النسيجية الكيميائية للكشف عن CD34 في خزع نقي عظم الحالات المرضية والمجموعة الضابطة. النتائج :- كان معدل كثافة الاوعية الدموية لابيضاض الدم اللمفاوي المزمن 10.78 والذي كان اعلى من معدل كثافة الاوعية الدموية الدقيقة للمجموعة الضابطة والذي كان 4.26 . (هذا الفرق معتد به احصائيا") كما وجدت علاقة عكسية ومعتدة بين زيادة كثافة الاوعية الدموية الدقيقة و كلا من مستوى حجم الخلية المحشو و عدد الصفيحات.هناك علاقة موجبة و معتدة احصائيا بين كثافة الاوعية الدموية و كلا من النسبة المئوية للخلايا اللمفاوية في نقي العظم, طراز ارتشاح نقي العظم وتقدم مرحلة المرض. الاستنتاجات :-1.تكون الاوعية الدموية الدقيقة في نقي عظم المرضى اعلى من المجموعة الضابطة.2.وجدت علاقة موجبة و معتدة احصائيا بين زيادة تكون الاوعية الدموية الدقيقة و مرحلة عالية الخطورة للمرض. 5. الدالة ٣٤- التعبير المناعي النسيجي - ابيضاض الدم اللمفاوي


Article
Angiogenesis and MMP-2expression in Oral Squamous Cell Carcinoma&Verrucous Carcinoma and its Correlation with Clinicopathological Parameters

Authors: Bashar H. Abdullah --- Seta A. Sarkis --- Ban F. AL Drobie
Journal: Diyala Journal of Medicine مجلة ديالى الطبية ISSN: 97642219 Year: 2014 Volume: 6 Issue: 1 Pages: 15-26
Publisher: Diyala University جامعة ديالى

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Abstract

Background: An important step of tumor progression in which Matrix metalloproteinase have been implicated is angiogenesis, because these enzymes degrade the extracellular matrix and provide a permissive microenvironment for the growth of new blood vessels.The present study conducted to evaluate the immunohistochemical expression ofMatrixmetalloproteinase -2(MMP-2) andangiogenic marker (CD34) in Oralsquamous cell carcinoma(SCC)versus verrucous carcinoma(VC) and to correlate their expressions with the clinicopathological parameters.Material & Methods: MMP-2 and CD34 expression was examined immunohistochemically in twenty four paraffin tissue blocks of squamous cell carcinoma and verrucous carcinoma (twelve cases of each). Results: All cases of Oral SCC exhibited positive immunostaining for MMP-2, while only one case of VC showed –ve expression. Interestingly all cases of VC showed –ve MMP-2 immunostaining of the basal cell layer. Generally lymphatic vessels were more than blood vessels in both VC&SCC cases. The mean MMP-2 immunoexpression was (59.00%) for both stage I &stage II, while the higher CD34 immuno expression was in stage I. The mean expression of MMP-2 was higher in well differentiated OSCCs, while for CD34 it was higher in poorly differentiated OSCCs followed by moderately differentiated, then well differentiated, however no statistically significant difference was found. Non- significant correlation was found concerning the expression of both markers for both lesions.Conclusion: No statistical correlation was found between MMP-2 expression and angiogenesis in OSCC and OVC.

Keywords

SCC --- VC --- MMP-2 --- CD34


Article
A Study Of Angiogenesis In Human Colorectal Tumors By Using Anti-Cd34 Antibody

Author: Nabil Salmo
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2007 Volume: 49 Issue: 4 Pages: 425-433
Publisher: Baghdad University جامعة بغداد

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Background: Angiogenesis plays a crucial role in tumorigensis; several reports have described a significant increase in microvessel density (MVD) in colorectal carcinogenesis There are several methods to measure the angiogenesis in neoplasms, but immunohitochemistry seems to be the mainstay of all. This method enable us to measure the tumor microvessel densities highlighted by using antibodies directed against endothelial cell markers like CD31,CD34 or others; then assessment of MVD by manual count of the number of microvessels in what appears to be the most vascular area of the tumor(called the hot spot) using a protocol described by Weidner et al.Automated cellular imaging system is used to analyze immunohistochemically stained slides .studies have shown that the device offers accurate precision and reproducibility of immunostained slide analysis exceeding that possible with manual evaluation which was the prevalent method . Aims of the study: To assess the angiogenesis in normal, adenomatous (benign) and malignant colorectal tissues using CD34 and the microvessels will be measured both manually by hot spot method as the MVD and by the use of computerized image analysis system as fraction area ,we correlate between microvessels density and fraction area with various clinicopathological parameters in colorectal cancer (CRC), and to a compare between the results which obtained from both methods. Methods: Paraffin embedded archival materials from 50 cases including three normal resection (non tumoruos) margins, 12 benign colonic lesions and 35 colonic adenocarcinoma were used . 5mm section were cut and they were stained by anti CD34 antibody Angiogenesis was measured as MVD by two methods : manually by light microscope and by a computer image analysis system (as fraction area). Then the MVD and fraction area were correlated with different clinicopathological parameters. Results : This study demonstrate that there is a statistical difference in MVD and fraction area in both hot spot method and CIAS respectively between benign and malignant tumors . P value < 0.05 in hot spot method and less than 0.001 in CIAS and there was highly significant correlation between MVD and fraction area with the grade . There was significant increase in MVD and fraction area from well differentiated to moderately differentiated and to poorly differentiated .There was no significant correlation between MVD and lymph node involvement by hot spot method but CIAS proved a significant correlation between fraction area and lymph node involvement.Both methods (hot spot and CIAS) proved no significant correlation with age , sex , size of the tumor , site of the tumor , stage of the tumor and the number of lymph node involverment . Conclusions: Assessment of tumor vascularity by counting the microvessels using anti CD34 antibody is useful in quantifying angiogenesis in colorectal adenocarcinoma . Intratumoral microvessels count (by manual and computerized method) is important in the assessment of the biological behavior of CRC. Microvessel count is higher in malignant tumors than benign tumors and it is correlated with the tumor grade and higher MVD is associated with lymph node metastasis . Statistically no significant correlation was found between MVD and age, sex of the patients, tumor size, site, stage, histological type and number of lymph node metastasis . The use of CIAS for assessment of angiogenesis is reliable, reproducible and more precise than the manual method.


Article
IMMUNOHISTOCHEMICAL EXPRESSION OF CD34, SMOOTH MUSCLE ACTIN AND TYPE IV COLLAGEN IN BREAST CARCINOMA. A CLINICOPATHOLOGICAL STUDY

Authors: Ban J Qasim بان جمعة قاسم --- Alaa G Hussein علاءغني حسين --- Rasha F Salih رشا فوزي صالح
Journal: IRAQI JOURNAL OF MEDICAL SCIENCES المجلة العراقية للعلوم الطبية ISSN: P16816579,E22244719 Year: 2012 Volume: 10 Issue: 2 Pages: 148 -153
Publisher: Al-Nahrain University جامعة النهرين

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Background: Tumoral angiogenesis is essential for the growth and spread of breast cancer cells.Objective: To evaluate angiogenesis by measuring microvessel density (MVD) with CD34 and its maturity with smooth muscle actin (SMA) immunohistochemistry and to study invasion of basement membrane by tumor cells using type IV collagen.Methods: In the present study microvascular quantification was undertaken on 52 cases of breast carcinoma and 5 cases of benign breast lesions after immunohistochemical staining of tumor vessel, using CD34 antibody and SMA antibody. Microvessel quantification was performed at x400 magnification in the three most vascular areas of the tumors (hot spots).Results: The difference in MVD between benign and malignant cases is significant (P= 0.001). MVD is significantly correlated with L.N. involvement (P=0.004) and lymphovascular permeation (P=0.001), no statistical significant correlation between MVD and age of patient (P=0.656), tumor size (P=0.052), tumor grade (P=0.324).Conclusion: Measurements of angiogenesis may have clinical utility in the evaluation of breast cancer, particularly for estimation of metastatic risk. A high MVD may be a poor prognostic marker of breast carcinoma and a target for antiangiogenic therapy.Key words: Angiogenesis, CD34, SMA, Collagen IV, breast carcinoma.

Keywords

Angiogenesis --- CD34 --- SMA --- Collagen IV --- breast carcinoma


Article
Assessment of microvessels density and inflammatory status in oral lichen planus

Author: Ban F. Al- Drobie بان فاضل الدروبي
Journal: Journal of baghdad college of dentistry مجلة كلية طب الاسنان بغداد ISSN: 16800087 Year: 2013 Volume: 25 Issue: special issue 1 Pages: 43-47
Publisher: Baghdad University جامعة بغداد

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Background: Oral lichen planus (OLP) is a chronic inflammatory disease with an autoimmune inflammatorypathogenesis. The purpose of this study was to evaluate the role of angiogenesis in the pathogenesis of OLP, usingCD34 stain to highlight the blood vessels for measuring the microvessel density (MVD) as well as to evaluate therelation of this marker with the degree of inflammationMaterials and Methods: Immunohistochemical (IHC) staining technique was used to evaluate angiogenesis usingCD34 in 46 paraffin blocks 10 of them obtained from normal mucosa and 36 from cases diagnosed as lichen planus ,20 of them diagnosed as Reticular type while 16 as erosive type. Severity of inflammation was divided into mild,moderate and severe according to the number of mononuclear cell infiltrate.Results: The mean MVD as determined by the mean number of CD34 positive vessels in hot areas for OLP group washigher 39.2 (31.9 for reticular group & 48.375 for erosive group), whereas the mean MVD for control group 19.5 so thatthere was statistically significant difference between OLP group and the control group (p value<0.001). Regardingthe severity of inflammation mean MVD was significantly increased as the degree of inflammation increased ( pvalue<0.001).Conclusion: Angiogenesis may be an integral component associated with the development of the OLP


Article
Assessment Of Vascular And Lymphatic Vessels Density In Benign Vascular Lesions Using CD34 And D2-40 Immunohistochemical Markers

Authors: Jawaher M. Tater --- Bashar H. Abdullah
Journal: Journal of baghdad college of dentistry مجلة كلية طب الاسنان بغداد ISSN: 16800087 Year: 2017 Volume: 29 Issue: 2 Pages: 61-64
Publisher: Baghdad University جامعة بغداد

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Background: Vascular tumors and malformations, comprising a broad category of lesions often referred to asvascular anomalies. Hemangioma, represents a variety of vascular lesions (both malformations and tumor), whilelobular capillary hemangioma is a common vascular lesion of the skin and mucous membranes that occurs mainly inchildren and young adults. Lymphangiomas are malformations of the lymphatic system. At the level of lightmicroscopy the small lymphatics vessels may be similar to capillaries and sometimes are only tentatively identified bythe nature of their contents or by immunohistochemical staining procedure. This study aimed to assess the vascularand lymphatic vessels density in benign vascular lesions using CD34 and D2-40 immunohistochemical markers.Materials and Methods: Twenty two formalin-fixed paraffin-embedded tissue blocks of Hemangioma/vascularmalformation, thirty of lobular capillary hemangioma and another twenty of lymphangioma.Results: Lymphatic vessel density expressed by D2-40 immunomarker was found in all cases with mean (24.01±14.74)in lymphangioma ,for lobular capillary hemangioma it was (12.67±6.66) and for hemangioma was (9.77±6.82)where as the mean of microvessel density count measured by CD34 immunomarker was (49.87±31.97) for lobularcapillary hemangioma , in hemangioma it was (37.42±23.40) and (25.90±12.23 ) for lymphangioma.Conclusions: All vascular lesions are a mixture of blood and lymphatic vessels with different proportions,hemangioma shows high percentage of blood vessels and lymphangioma shows high percentage of lymphaticvessels.


Article
Trehalose and ascorbic acid improves the Cryopreservation of umbilical Cord Blood hematopoietic stem Cells (CD34+) with low Concentrations of Dimethylsulfoxide
التريهالوز وحامض الاسكوربيك يحسنان الخزن بالتجميد لخلايا دم الحبل السري الجذعية + CD34 مع التراكيز القليلة لـ Dimethylsulfoxide

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Abstract

Cryopreservation of umbilical cord hematopoietic stem cells (HSCs) is essential step in stem cell transplantation. TheDimethylsulfoxide (DMSO) mostly used as a cryoprotective agent, associated with toxic effects to stem cells. Inorder to minimize the effects of DMSO, low concentrations of DMSO with additional cryoprotectants should used. In thisstudy mononuclear cells (MNCs) isolated by ficoll from umbilical cord blood (UCB), which contain hematopoietic stemcells (CD34+), were used for cryopreservation.Cryopreservation of UCB-derived MNCs was done for period of one month by using uncontrolled-rate freezingtechnique at -196˚C in liquid nitrogen. Cryopreservation solution was used which consisted of minimum essentialmedium (MEM) and 20% fetal calf serum (FCS) supplemented with the cryoprotectant dimethylsulfoxide (DMSO) intwo concentrations 2.5% and 5% DMSO, lower than usually used 10%, with and without 25μg/ml trehalose or 80μg/mlascorbic acid to improve cryopreservation process. The addition of trehalose and ascorbic acid improved cryopreservationprocess in comparison with control. Addition of 5% DMSO alone and with additives showed a better result than 2.5%DMSO alone or with additives for cryopreserving CD34+ cells as indicated by immunocytochemistry.Washing out DMSO also affected the count and viability of MNCs. These results indicated that it could use lowconcentrations of DMSO in cryopreservation of HSCs by association with trehalose and ascorbic acid.

عادة Dimethylsulfoxide (DMSO) الخزن بالتجميد لخلايا الحبل السري الجذعية خطوة اساسية في نقل الخلايا الجذعية. اليستخدم تراكيز اقل DMSO يستخدم كعامل حماية من التجميد يرافقه بعض السمية للخلايا الجذعية. لاجل تقليل تأثير المنه مع مواد تحمي من التجميد. في هذه الدراسة تم عزل الخلايا وحدة النواة والحاوبة على الخلايا الجذعية باستخدام الفيكول لغرض˚ التجميد. تم النجميد لخلايا وحيدة النواة والمعزولة من دم الحبل السري لمدة شهر باستخدام تقنية التجميد الممباشر بدرجة - 196 م% و 20 % من سيرم جنين الابقار بالاضفة الى تراكيز اقل من 10 MEM بالنايتروجين السائل. يتألف محلول الخزن بالتجميد من وسطوهي 2.5 % و 5% مع و بدون 25 مايكروغرام/مل من التريهالوز أو 80 مايكروغرام/ مل من حامض الاسكوربيك لتحسين DMSO منعملية الخزن بالتجميد. ان اضافة التريهالوز وحامض الاسكوربيك حسن عملية التجميد بالقارنة بالسيطرة.لوحده او مع الاضافات اظهرت نتائج افضل من تركيز 2.5 % لوحده او مع الاضافات للخلايا الجذعية المخزونة DMSO % ان اضافة 5أثر على عدد وحيوية الخلايا الوحيدة النواة. هذه النتائج اكدت DMSO بالتجميد كما اثبت بتقنية الكيمياء المناعية. غسل الفي الخزن بالتجميد للخلايا الجذعية باضافة التريهالوز وحامض الاسكوربيك. DMSO امكانية استخدام تراكيز اقل من


Article
Immunohistochemical study of bone marrow angiogenesis in multiple myeloma and its correlation with clinicopathological factors
دراسة مناعية نسيجية كيميائية لعملية تكون الاوعية الدموية الجديدة لنخاع العظم في الورم النقوي المتعدد وعلاقتها مع مختلف المعايير المرضية والسريرية

Authors: Rana Z. Naji رنا زهير ناصي --- Abdulkareem M. Jaafar عبد الكريم محمد جعفر --- Hasan A. Buraid حسن عبد الرزاق بريد
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: 00419419 Year: 2014 Volume: 56 Issue: 2 Pages: 195-199
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: In recent years, bone marrow angiogenesis was reported to be involved in the pathogenesis and progression of certain hematological malignancies like multiple myeloma, leukemias, and lymphomas. Recent studies have suggested that bone marrow angiogenesis plays an important role in the pathogenesis and prognosis of multiple myeloma.Objectives: at the present study, bone marrow angiogenesis in multiple myeloma was examined using immunohistochemical staining for CD34, and correlated with various pathological and clinical parameters.Patients and methods: This is a retrospective study; where by archival paraffin-embedded tissue blocks along with the clinical and hematological records of fifty-two patients with multiple myeloma and twenty controls were obtained from the Department of Hematology of the Medical City Teaching Laboratories in the period from January 2010 to January 2013. Bone marrow angiogenesis was studied by immunohistochemical staining for CD34 to identify microvessels.Results: There was a significant association between higher angiogenesis grades and advanced clinical stage of the disease (p = 0.002), and higher plasma cell percent in the bone marrow (p = 0.03), and increasing immaturity of the plasma cell (p = 0.001), and diffuse pattern of bone marrow infiltration by plasma cells (p = 0.007).Conclusions: Patients with increased plasma cell burden, immature plasma cell morphology, and diffuse pattern of infiltration had a higher microvessel density. Key words: Multiple myeloma; angiogenesis; CD34

خلفية عن الدراسة: في السنوات الاخيرة اصبح من الضروري معرفة عملية تكون الاوعية الدموية الجديدة في نخاع عظم المريض لفهم عملية تطور المرض وتكهن نتائجه الاخيرة خصوصا في بعض سرطانات الدم مثل الورم النقوي وسرطان الدم وسرطان العقد اللمفاوية. إفترضت الدراسات الحديثة ان عملية تكون الاوعية الدموية الجديدة في نخاع العظم تلعب دور مهم في عملية تطور المرض وتكهنه لدى المصابين بالورم النقوي.الهدف من الدراسة: فحص عملية تكون الاوعية الدموية الجديدة لدى المرضى المصابين بالورم النقوي المتعدد باستعمال التقنية المناعية النسيجية الكيميائية للدالة CD34 وعلاقتها مع مختلف المعايير المرضية والسريريةالمرضى والمواد واطرائق العمل: هذه دراسة بأثر رجعي حيث تحلل قطع نخاع العظم المغمورة في البارافين والموجودة في الارشيف مع السجلات السريرية وسجلات تحاليل الدم الى 52 مريض مصاب الورم النقوي المتعدد مع عشرين فرد سيطرة تم الحصول عليها من قسم امراض الدم في المختبرات التعليمية في مدينة الطب والمسجلة من كانون الثاني 2010 الى كانون الثاني 2013. تمت دراست عملية تكون الاوعية الدموية الجديدة في نخاع العظم باستخدام تقنية الصبغ المناعية النسيجية الكيميائية للدالة CD34 لتمكننا من التعرف على أدّق الاوعية الدموية.النتائج: كانت هنالك علاقة ترابط مميزة ما بين درجات عملية تكون الاوعية الدموية الدقيقة الجديدة العالية وبين المرحلة السريرية المتقدمة للمرض (P=0.002) وبين النسبة العالية لخلايا البلازما في نخاع العظم (P=0.03) وبين عدم النضوج الزائد لخلايا البلازما (P=0.001) وبين النمط المنتشر لتسرب خلايا البلازما في نخاع العظم (P=0.007).الإستنتاجات: المرضى الذين لديهم زيادة في نسبة خلايا البلازما او زيادة عدم النضوج الشكلي او زيادة النمط المنتشر لتسرب خلايا البلازما في نخاع العظم تكون لديهم كثافة عالية للاوعية الدموية الدقيقة.كلمات مفتاحية: الورم النقوي المتعدد ;عملية تكون الاوعية الدموية الدقيقة الجديدة ;الدالة CD34.

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