ORIGINAL ARTICLE

Immunohistochemical Evaluation of Sentinel Lymph Nodes in Colon CancerSorlea S.1, Coros M.F.2, Georgescu R.2, Gyorgy-Fazakas I.1, Branzaniuc Klara1, Milutin Doina3, Pavai Z.1, Copotoiu C.4

1 Discipline of Anatomy and Embriology, Faculty of Medicine, University of Medicine and Pharmacy, Targu Mureș 2 Clinic of Surgery I, Faculty of Medicine, University of Medicine and Pharmacy, Targu Mureș, Mureș County Hospital 3 Pathology department, Faculty of Medicine, University of Medicine and Pharmacy, Targu Mureș 4 Clinic of Surgery I, Faculty of Medicine, University of Medicine anf Pharmacy, Targu Mureș, Emergency County Hospital

Background: Lymph node status is the most important predictive factor in the treatment of colic cancer. As sentinel lymph node (SLN) biopsy might upstage stage II colon cancer, it could have therapeutic consequences in the future. Aim: To investigate and evaluate nodal microstaging and ultrastaging using cytokeratin immunohistochemistry. Material and methods: In 20 consecutive patients operated on First Surgery Clinic of the County Hospital Mures for colon cancer, subserosal injection with Patent Blue dye was used for SLN detection. In searching for occult micrometastases, each SLN was ex-amined. In tumor-negative SLNs at routine hematoxylin-eosin (H&E) examination (pN0) we performed cytokeratyn(CK) immunohisto-chemistry (IHC). Results: The procedure was successful in 19 out of 20 patients (95%). The SLN was negative in 12 patients detected by H&E and IHC, in 10 patients the non-SLN was also negative, leading to a negative predictive value of 89% and an accuracy of 93%. In 6 patients with SLN negative by HE was pozitive by IHC, leading to a 33% value of upstaging.Conclusions: The SLN concept in colon carcinoma using Patent Blue V is feasible and accurate. It leads to upstaging of nodal status in 6 cases (33%) when IHC techniques are involved. The clinical value of the method will be evaluated by postoperative chemotherapy efficiency.

Keywords: sentinel lymph node, colon, carcinoma

IntroductionColorectal carcinoma is the most common gastrointesti-nal malignancy. Lymph node status as the most important predictor of outcome indicates the use of adjuvant che-motherapy.The reported 5-year survival rate is 70–80% for patients with node negative disease (st. I–II), but only 45–50% for those with node positive tumors (st.III) [1]. Adjuvant chemotherapy significantly improves the 5-year survival in patients with node positive disease. Despite the favorable prognosis of patients with localized colon cancer without regional lymph node metastasis, 20–30% of these patients will develop recurrent disease, after apparently cu-rative resection. It is therefore necessary to perform a more detailed histological examination of negative lymph nodes by histological examination with haematoxylin-eosine staining (HE) and immunohistochemistry with cytoker-atin (CK).Understaging may be the result of inadequate numbers of examined lymph nodes, missing some metasta-ses [1,2,3]. For adequate staging and treatment of patients with colon cancer, meticulous examination of at least 12 nodes harvested by pathological analysis is mandatory [4].

Sentinel node technique was described by Cabanas in 1977 in penile cancer, and Morton Giuliano introduced the method for melanoma and breast cancer. In colon can-cer the sentinel lymph node is defined as the first tumor draining lymph node, with the highest potential to harbor metastatic disease [5,6,7]. This allows a targeted examina-tion of a smaller number of nodes that can be examined

with multiple sections and immunohistochemistry for the accurate detection of metastases and micrometastases and to provide a better staging of colon cancer.

We used methylene blue dye to identify sentinel lymph nodes and examined them with haematoxylin-eosine stain-ing and immunohistochemical technique with cytokera-tin. In tumor-negative SLN’s at routine hematoxylin-eosin (H&E) examination (pN0) we performed CK8/CK18 im-munohistochemistry (IHC).

Material and methodsOnly patients with histological proven primary colon car-cinoma were included in the study. Patients with distant metastases or gross lymph node involvement as shown by preoperative examinations or palpation during surgery were excluded.

Sentinel lymph node mapping was carried out through an open procedure by injection of 1–3 ml Blue Dye with a tuberculin syringe and 29 gauge needle subserosally in 4 quadrants around the tumor. The subserosal injection was carried out prior to vascular ligation. Within 5 to 10 min-utes after the blue dye injection, the SLN’s could be identi-fied by following blue stained lymphatic vessels leading to the blue stained sentinel node [8–12]. These nodes were tagged with a long suture. Sentinel nodes were defined as the first four bluestaining nodes seen within the regional basin. After marking of the SLN’s, routine resection was performed.

BIOPSIA NODULULUI SANTINELA IN CANCERUL DE COLON

S.SORLEA1, M.F.COROS2, R.GEORGESCU2, I.GYORGY-FAZAKAS1, KLARA BRANZANIUC1, DOINA MILUTIN3, Z.PAVAI1, C.COPOTOIU4

1- Disciplina Anatomia si Embriologie, Universitatea de Medicina si Farmacie, Targu Mureş 2- Clinica Chirurgie 1, Spitalul Clinic Judetean Mures, Universitatea de Medicina si Farmacie, Targu Mureş 3- Departmentul de AnatomoPatologie, Universitatea de Medicina si Farmacie, Targu Mureş 4- Clinica Chirurgie 1, Spitalul Clinic Judetean de Urgenta Tg-Mures, Universitatea de Medicina si Farmacie, Targu Mureş

Rezumat Introducere:Statusul nodulilor limfatici este cel mai important factor predictiv în tratamentul cancerului de colon. Biopsia nodulului santinelă poate trece din stadiul I în stadiul II cancerul de colon, cu consecinţe asupra tratamentului postoperator . Chimioterapia adjuvantă administrată în cazurile cu invazie ganglionară îmbunătăteşte semnificativ supravieţuirea. Material si metoda:Am utilizat injectarea subseroasă peritumorala de colorant albastru de metilen pentru identificarea SLN la o serie de 40 pacienţi operaţi pentru cancer de colon în Clinica Chirurgie 1 a Spitalului Clinic Judeţean Mures.pentru detectarea metastazelor am utilizat examinarea histologica cu coloratie hematoxilin eozina(HE), În cazul nodulilor sentinela negativi (pN0) la coloraţia de rutina cu hematoxilin eozină s-a realizat examinarea imunohistochimică cu citokeratină (IHC). Rezultate:Am realizat procedura cu succes la un număr de 38/40 pacienţi (95 %). Un numar total de 530 de noduli limfatici au fost examinati,dintre acestia au fost marcati ca si noduli sentinela un numar de 76 (in medie 1,89/caz). SLN a fost negativ la un numar de 24 pacienţi atît la coloraţia HE cît şi la IHC, la 20 pacienţi ganglionii non SLN au fost deasemenea negativi cu o valoare predictiv negativă de 83,33 % şi o acurateţe de 93%. La 9 pacienţi cu SLN negativ la coloraţia HE , au fost pozitivi la IHC ceea ce a condus la o suprastadializare de 23,68 %. Concluzii:Utilizarea examinarii ganglionului santinelă si identificarea acestuia utilizând albastru de metilen este o metodă fezabilă şi corectă. Am realizat suprastadializarea prin utilizarea examinării imunohistochimice (IHC) în 9 cazuri (23,68 %). Biopsia nodulului santinelă poate trece din stadiul I în stadiul II cancerul de colon, cu consecinţe asupra tratamentului postoperator. Valoarea clinică a metodei va fi evaluată prin eficienţa tratamentului chimioterapic postoperator la acesti pacienti.

210 Clujul Medical 2011; 84, Supl. 2 - Vol.1

Al XII-lea Congres al Societății Române de Anatomie, Cluj-Napoca 9-11.06.2011

KT.pdf RMF­OGS .pdf(Digitized )

AMM.pdf CM.pdf Ratios of values from Articles

TitlesLymph node 

staging in colon cancer 

(chap. 4)

The role of sentinel node in the treatment of 

colon cancer

Immunohistochemical Evaluation of SLN in Colon 

Cancer

Biopsia nodulului santinela in cancerul 

de colon

Article 2/Article 4; 

Article 1/Article 4

Authors Kelder, Wiggers

Sorlea, Copotoiu et al

Sorlea, Coros et al

Sorlea, Coros et al (small  integers)

Total patient number 30 20 20 40 1/2;  3/4Range of ages of all patients 48­85 49­79 49­79 49­79 identity; ­Average  age of all patients 69 61.45 61.45 61.45 identity; ­Number of women 14 12 12 24 1/2; ­Average age of women ­ 60.91 60.91 60.91 identity; ­Number of men 16 8 8 16 1/2 ;  identityAverage age of men ­ 62.25 62.25 62.25 identity; ­Total number of patients with SLN 29 19 19 38 1/2; ­Successful of procedure 97% 95% 95% 95% identity; ­Number of failed cases by procedure 1 1 1 2 1/2;   1/2Number of nodes/sample ­ 3 ­ 32 3 ­ 32 3 ­ 32 identity; ­Mean number of nodes/sample ­ 13.75 13.75 13.75 identity; ­Procedure accuracy 93% ­ 93% 93% ­ ; identityNegative predictive value 89% ­ 89% 83.33 ­ ;   ­Value of upstaging 33% ­ 33% ­ ­ ;   ­Patients with positive SLN by H­E  4 6 6 12 1/2;   1/3; 2/3Percentiles of patients with positive SLN ­ 30% 30% 30% identity;  ­Number of patients with SLN+ & other LN+

­ 6 6 12 1/2; ­

Percentiles patients SLN+ & other LN+ ­ 30% 30% 30% identity; ­Number of patients with SLN­ (H­E & IHC)

18 ­ 12 24 ­;   3/4

Abbreviations: RMF­OGS, Revista de Medicină şi Farmacie­Orvosi és Gyógyszerészeti Szemle; AMM = Acta Medica Marisiensis; CM, Clujul Medical; SLN, Sentinel Lymph Nodes; H­E, Hematoxylin­Eosin; IHC, Immunohistochemistry.

ORIGINAL ARTICLE

IntroductionThe cartilage is a complex and specialized tissue. It is ex-tremely difficult to repair or to replace it, once damaged. The repair tissue found in the cartilage defects is fibrocar-tilage, which is mechanically and chemically inferior to hyaline cartilage [1]. The management of cartilage defects remains controversial and over the last five decades various treatment options and surgical techniques have been tried to optimize the clinical outcome.

In a review of 993 knee arthroscopies in patients with a mean age of 35 years, there was an 11% incidence of full-thickness lesions that could have benefited from surgical treatment [2]. In a larger and more generalized study, Curl et al. reviewed 31,516 knee arthroscopies of patients in all age groups and reported chondral lesions in 19,827 (63%) of patients; 5% of all cases were found in patients younger than 40 years of age who had grade IV lesions [3]. A review of 1,000 arthroscopies by Hjelle et al reported chondral or osteochondral lesions of any type in 610 patients (61%), out of which 190 patients had focal lesions (19% of all cases). Many of these lesions were clinically silent at the time of detection [4].

Keeping in mind that those procedures are relatively new, we presented the author’s and the Orthopedic Clinic's experience in using these techniques. The aim of this study is to evaluate, but not compare, the results of two of the

most used cartilage repair techniques: transchondral drill-ing and osteochondral autografting.

Material and methodsBetween January 2009 and June 2010, we performed 55 transchondral drillings and 10 mosaicplasties on patients with articular cartilage defects of the knee. The study is a prospective longitudinal one, with 6 months patient fol-low-up. In the group with transchondral drilling, 39 pa-tients (70.9 %) were male. In the group with mosaicplasty, 8 patients were male and 2 female. The medial condyle was affected in 58 cases (89.23%).

The mean age for the group with transchondral drilling was 42.55±9.32 years, the patients being between 19 and 49 years old. For the group with mosaicplasty, the mean age was 44.23±6.87 years, the patients being between 39 and 51 years old.

In all patients we performed a conventional radiogra-phy (anteroposterior and lateral views). In 8 patients, 6 from the transchondral drilling group (10.9%) and 2 pa-tients from the mosaicplasty group we performed a CT scan. Magnetic resonance imaging was performed in 13 patients from the transchondral drilling group (23.6%) and 4 patients from the mosaicplasty group.

The performed procedure was chosen based on patient age, physical activity and lesion size.

Transchondral Drilling and Osteochondral Autografting (Mosaicplasty) in Knee Articular Cartilage DefectsIvănescu A1, Melinte R2, Sólyom Á2, Moraru L3, Petrișor M4, Brânzaniuc Klara1

1 University of Medicine and Pharmacy of Tîrgu Mureș, Department of Anatomy and Embriology 2 County Emergency Clinical Hospital Tîrgu Mureș, Orthopedic Clinic II 3 County Emergency Clinical Hospital Tîrgu Mureș, Clinic of Cardiovascular Surgery 4 University of Medicine and Pharmacy of Tîrgu Mureș, Department of Medical Informatics and Biostatistics

Background: The cartilage is a complex and specialized tissue. It is extremely difficult to repair or to replace it, once damaged. The management of cartilage defects remains controversial and over the last five decades various treatment options and surgical techniques have been tried to optimize the clinical outcome. Objective: The aim of this study is to evaluate, but not to compare the results of two of the most used cartilage repair techniques: trans-chondral drilling and osteochondral autografting. Material and methods: Between January 2009 and June 2010, we performed 55 transchondral drillings and 10 mosaicplasties on patients with articular cartilage defects of the knee. All patients were followed up at 6 months. Hughston clinical and radiological scales were used to evaluate the patients in the transchondral drilling group. Results: The Hughston Clinic score was 2 in 2 cases (3.6%), 3 in 5 cases (9.9%) and 4 in 48 cases (86.5%), giving over 95% of good results. The Hughston radiological score was 2 in one case (2%), 3 in 4 cases (7.3%) and 4 in 50 cases (90.7%). In the mosaicplasty group,the average area of the osteochondral lesion covered with autologous osteochondral transplantation ranged from 0.8 to 6 cm2 (average: 2.13 cm2). The diameter of the grafts used ranged from 6 to 10 mm and 1 to 6 grafts were used in each case to achieve >90% covering of the lesion area. Conclusions: Both techniques offer satisfactory functional outcome and do not compromise the patients’ future options.

Keywords: transchondral drilling, osteochondral autografting, mosaicplasty, articular cartilage

RESEARCH ARTICLE

Acta Medica Marisiensis 2013;59(2):165-168

The New Changes in the 7th AJCC/UICC Staging System of Gastric CarcinomasHălmaciu Ioana1, Gurzu Simona2, Suciu BA3, Comișel SI4, Dénes L1, Boc Lacrima5, Brînzaniuc Klara1

1 Department of Anatomy and Embryology, University of Medicine and Pharmacy, Tîrgu Mureș, Romania2 Department of Pathology, University of Medicine and Pharmacy, Tîrgu Mureș Romania3 Department of Surgery, University of Medicine and Pharmacy, Tîrgu Mureș, Romania4 Department of Oral and Maxillofacial Surgery, County Emergency Clinical Hospital, Tîrgu Mureș, România5 Student, University of Medicine and Pharmacy, Tîrgu Mureș, Romania

Objective: The aim of this study was to analyze in parallel the 6th and the newest 7th AJJCC/UICC (American Joint Committee on Cancer/

International Union Against Cancer) staging system in order to highlight changes brought by the new staging system.

Methods: We analyzed data obtained retrospectively from 134 hospitalized patients diagnosed with gastric carcinomas, who underwent

surgery at the Surgery Clinics of the County Emergency Clinical Hospital of Tîrgu Mureș, Romania between 2008–2010. The data have been

obtained from histopathology reports, and the analyzed parameters were the following: age, gender and pTNM staging. For all cases included

in the study restaging was performed according to the 7th AJJCC/UICC staging system.

Results: 71.66% of cases were adenocarcinomas, 7.46% mucinous adenocarcinoma, 14.17% signet ring cell carcinoma, and 6.71% undif-

ferentiated carcinoma. The signet ring cell carcinomas predominated before 65 years of age (p = 0.003). Compared to the 6th staging system,

in the new system pT2 percentages decreased signifi cantly from 38.8% to 6.71%, and pT4 increases from 11.19% to 55.97% (p <0.0001).

The pN3 cases increased from 20.9% to 45.52%, because all cases classifi ed as pN2 in the old staging system, became pN3 in the new

system. Some of the pN1 cases turned into pN2 in the new system (p = 0.004). Stage IV cases also decreased from 29.85% to 14.94%, due

to regrouping of stage III.

Conclusions: There are signifi cant changes between the two staging systems. The new staging system aims to achieve a better postopera-

tive follow-up.

Keywords: gastric cancer, 6th AJJCC/UICC staging system, 7th AJJCC/UICC staging system

Received: 27 April 2012

IntroductionGastric cancer is one of the most frequent gastrointestinal tumors, and represents the second cause of cancer death worldwide, although global incidence is declining [1].

It is known that most of gastric cancer patients are di-agnosed in advanced stages, due to unspecifi c symptoms, and also to late reporting of patients to the physician [2]. Surgery is the only option providing substantial improve-ment of survival in cases with early diagnosis, but even in patients diagnosed with early stages, the 5-year survival rate is about 50% [3]. Patients with advanced stages of gastric cancer can benefi t from palliative care or neoadju-vant chemotherapy. Th us, accurate quantifi cation of tumor stage is an extremely important aspect in establishing the subsequent treatment protocol for the patient.

Th e stage of the disease also represents one of the most important prognostic factors of gastric cancer; therefore TNM staging has the main role in establishing the treat-ment protocol [4]. In 2010 the 7th TNM staging of gastric carcinomas has been introduced by the American Joint Committee on Cancer (AJCC) and International Union Against Cancer (UICC) [5].

Th e present study aims to highlight the importance of changes brought by the 7th TNM staging of AJCC/UICC, in order to achieve a better postoperative staging. Th e changes brought by the 7th staging system compared to the 6th staging system are listed in Table I [5,6].

MethodsOne-hundred forty-three patients who underwent surgical intervention during 2008–2010 were enrolled in the study. Open surgery was performed in each case to remove the gastric tumor. In all cases, formalin-fi xed embedded tissues were used. Sections were dewaxed and were stained with Hematoxylin-Eosin.

We analyzed the histological type and grade of the tu-mor. Th ese parameters were correlated with the patients’ age and gender. Only carcinomas of the stomach were in-cluded in our study. Lymphomas, carcinoid tumors and gastrointestinal stromal tumors were excluded.

We analyzed in parallel the 6th and 7th AJCC/UICC staging systems [5,6] in order to underline the clinical sig-nifi cance of the new staging of gastric carcinomas.

Data was collected with Microsoft Excel, and analyzed with GraphPad InStat software. Categorical data analysis was conducted with the chi-square test. Th e level of signifi -cance was set at p <0.05.

Correspondence to: Ioana Hălmaciu

E-mail: anca_halmaciu@yahoo.com

DOI: 10.2478/amma-2013-0039

Unauthenticated | 86.126.25.127Download Date | 10/28/13 8:35 PM

ORIGINAL ARTICLE

Stem Cells Harvest from Volunteer DonorsTănase Alina1, Berteanu Cristina2, Călugăroiu Carmen1, Marculescu Alexandra1, Copotoiu Sanda-Maria3, Brânzaniuc Klara3, Azamfirei L3, Cirstoiu C2, Stoica Maria4, Cernea Daniela4, Stoica GA4, Rosin A5

1 Fundeni Bone Marrow Transplant Center Fundeni 2 Bucharest Emergency University Hospital 3 University of Medicine and Pharmacy of Tîrgu Mureș 4 Craiova Emergency University Hospital 5 Bucharest Institute of Haematology

Background: Over the last several decades allogeneic hematopoietic cell transplantation (HCT) has emerged as an important therapeutic option for a number of malignant and non-malignant conditions. The collection of hematopoietic stem cells mobilized from the bone marrow into the bloodstream of healthy donors has now become a routine procedure throughout the world.Materials and methods: A number of 86 procedures of hematopoietic stem cells (HSC) harvest and cryopreservation from 64 volunteer donors, 54 adults (28 women and 26 men) and 10 children (5 girls and 5 boys) with ages between 6–66 years (on ave-rage 30.5) were car-ried out in the Bone Marrow Transplant Center from Clinical Institute Fundeni, Bucharest.Results and discussions: HSC mobilization was achieved for all the 64 volunteer donors by administration of Filgastrim, on an average 8.4 mcg/donor weight (limits: 5–16.64 mcg/donor weight), leukapheresis procedure being realized in day +5 of Filgastrim administration.Conclusions: In conclusion, a healthy volunteer donor, will undergo in most cases 4 or 5 days of Filgastrim administration. The WBC and the number of CD34+ cells from the periphe-ral blood will be counted beginning with the 4th day. When the number of CD34+ cells from peripheral blood will reach a certain level (usually on the 4th or 5th day), the vo-lunteer donor will be sent to the apheresis unit for harvesting stem cells.

Keywords: stem cells, harvesting, volunteer donors

IntroductionWidely accepted cancer treatment strategies include chemo-therapy and radiotherapy. The rationale for administration high-dose chemotherapy and/or radiation to patients with therapy-sensitive tumor is to reduce tumor burden. Deli-very of these therapies with respect to higher drug doses and intensified schedule are often limited by organ toxicity (eg bone marrow, heart, lung) [1]. To overcome these dose limitations, hematopoietic stem cell transplantation, high-dose therapy supported by the infusion of hematopoietic stem cells, has evolved as a medical procedure to allow for administration of intense drug doses with tole-rable organ and hematopoietic toxicity [2].

Over the last several decades allogeneic hematopoietic cell transplantation (HCT) has emerged as an important therapeutic option for a number of malignant and non-ma-lignant conditions. Besides the effect of high-dose chemo-therapies and/or total body irradiation on malignancies the benefit of allogeneic HCT is due to the graft-versus-leukemia/tumor (GVL) immune reaction by transplanted donor immune cells. However, allogeneic HCT is limited by the immunologic recognition and destruction of host tissues, termed graft-versus-host-disease (GVHD). GVHD continues to be a major source of morbidity and mortality following allogeneic HCT which limits use of HCT for a broader spectrum of diseases and patients [3].

The collection of hematopoietic stem cells mobilized from the bone marrow into the bloodstream of healthy do-nors has now become a routine procedure throughout the world. Peripheral blood stem cells (PBSCs) mobilized by

recombinant human granulocyte-colony stimulating factor (rhG-CSF) were usually used for allogeneic hematopoietic stem cell transplantations. The immediate side effects of rhG-CSF administration and PBSC collection have long been established; however, information as to possible long-term consequences has been limited up to now [4].

Transplantation of G-CSF-mobilized peripheral blood cells is our current standard practice in allogeneic trans-plantation.

Pluripotent stem cells express the cell surface marker Ag CD34. This marker is the indicator most frequently used in clinical practice to determine the extend and efficiency of peripheral blood stem cells collections.

Although not a complete measure of quantity and qual-ity of collected cells, blood samples from collections are assayed to determine the number of CD34+ cells present. Once specific cell targets are achieved, cell collections are completed and stored for future use.

It is not known the minimum safe number of CD34+ cells needed for clinical engraftment of all lineages, as this may vary depending on the stem and progenitor cell subset composition in a given patient. However, it is known that a graft content of more than 5–10 × 106 CD34+cells/kg of body weight is safe, and, most important, only has a minor risk of engraftment failure [5].

Traditionally, HSC were harvested from the iliac crests under general anesthesia. Thereafter, mobilized PBSC have been increasingly used in both auto- and allo-HSCT. In the 1990s, unmanipulated CB cells collected and cryopre-served at birth have been used both in related and unrela-