Thromboelastographic profiles as a tool for thrombotic risk in digestive tract cancer
Background: Quantification of the magnitude of thrombotic risk associated with malignancy and with anti-cancer therapy is indispensable to use anticoagulant drugs which selectively interfere with haemostatic mechanisms protecting patients from venous thromboembolism (VTE) and probably from tumor pro...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2007
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Цитувати: | Thromboelastographic profiles as a tool for thrombotic risk in digestive tract cancer / M.L. Papa, F. Capasso, L. Pudore, S. Torre, S. Mango, V. Russo, P. Delrio, R. Palaia, F. Ruffolo, M.D. d’Eufemia, D. De Lucia, M. Napolitano, P. Di Micco, V. Parisi // Experimental Oncology. — 2007. — Т. 29, № 2. — С. 111-115. — Бібліогр.: 26 назв. — англ. |
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irk-123456789-1385812018-06-20T03:03:21Z Thromboelastographic profiles as a tool for thrombotic risk in digestive tract cancer Papa, M.L. Capasso, F. Pudore, L. Torre, S. Mango, S. Russo, V. Delrio, P. Palaia, R. Ruffolo, F. d’Eufemia, M.D. De Lucia, D. Napolitano, M. Di Micco, P. Parisi, V. Original contributions Background: Quantification of the magnitude of thrombotic risk associated with malignancy and with anti-cancer therapy is indispensable to use anticoagulant drugs which selectively interfere with haemostatic mechanisms protecting patients from venous thromboembolism (VTE) and probably from tumor progression. However, none of activation coagulation markers has any predictive value for the occurrence of the thrombotic events in one individual patient. Current clotting methods can’t reveal the overall dynamic clot formation; in contrast thromboelastographic methods specifically assess overall coagulation kinetics and its strength in whole blood. Aim: Objective of study was to evaluate if the activation of coagulation as eventually revealed by ROTEM® thromboelastometry could assess an hypercoagulable state in surgical neoplastic patients. Patients and Methods: Fifty consecutive patients with carcinoma of the digestive tract in preoperative period (23 M, 27 F aging 61.5 (45–79 years) and 147 healthy subjects (71 M, 76 F) were studied. A recent thromboelastometric method based on thrombelastography after Hartert was employed. Measurements were performed on ROTEM Coagulation Analyzer. The continuous coagulation data from 50 min course were transformed into dynamic velocity profiles of WB clot formation. Results: Standard parameters (CT, CFT, MCF) of cancer patients were similar to controls. CT (in cancer patients): females 50 s (38.3–58.7), males 50 s (42–71.2) vs 51 s (42–59), p = 0.1210 / 53 s (42–74.8), p = 0.1975 (in controls). CFT (in cancer patients): females 72 s (32- 92.4), males 80 s (50.2- 128.7) vs 78 s (62–100), p = 0.0128 / 80 s (59–124.4), p = 0.9384 (in controls). MCF (in cancer patients): females 70 mm (59.9–82.5), males 63 mm (56–73.7) vs 69 mm (59–95.8), p = 0.9911 / 69 mm (53.6–90), p = 0.0135 (in controls). Females showed a higher MaxVel when compared to males. The MaxVel was increased in cancer patients: females 19 mm /100 s (14.3–49.5) males 18 mm / 100 s (11–27) vs 15 mm 100 s (11.8–22), p < 0.001 / 13 mm / 100 s (10–21.8), p < 0.001 in controls .The t-MaxVel was shortened in cancer patients: females 65 s (48.6–112.8), males 81 s (50.1–135.9) vs 115 s (56.8–166), p <0.001 / 115 s (59.8–180.8), p = 0.0002 in controls. The AUC was increased in cancer patients: females 6451 mm 100 (5511–8148), males 5984 mm 100 (5119-6899) vs 5778 mm 100 (4998–6655), p < 0.001 / 5662 mm 100 (4704–6385), p = 0.0105. Conclusion: Unlike other assays measuring variations in a single component during coagulation, the thrombelastographic method records a profile of real-time continuous WB clot formation, and may provide extensive informations on haemostasis in neoplastic patients before surgery. Предпосылки исследования количественная оценка риска тромбоза, связанного со злокачественными заболеваниями и противоопухолевой терапией, обязательно включает в себя применение средств-антикоагулянтов, защищающих больного от развития венозной тромбоэмболии (VTE)и возможно п рогрессии заболевания . Тем не менее ни один из маркеров ак- тивации коагуляции не имеет прогностической ценности с точки зрения возможности возникновения тромбоза у каждого отдельно взятого пациента. Современные мето ды оценки свертывания крови не отража ют образование тромба винамике ; наоборот, метод тромбо эластографии дает возможность специфически оценить кинетику свертывания крови целом . Цель: определить, в какой мере активность коагуляции, определяемой методом тромбоэ ластометрии, отражает состояние гиперсвертываемости крови у больных онкологического профиля после хирургического вмешательства. Пациенты и м ды: обследованы 50 больных раком пищ еваритель ного тракта в дооп ерационный п ериод (27 женщин, 23 му жчины, средний возраст 61,5 года (45–79 лет) и 147 здоровых доноров (71 мужчина, 76 женщин). Применяли метод тромбоэластометрии , основанный на тромбоэластографии Гартерта, с использованием анализатора коагуляциифирмыROTEM. Текущие д анные о свертывании за 50 мин измерений представили в виде динамичных профилей вязкости при образовании сгустка крови. Результаты: стандартные параметры (перио д коагуляции (CT), перио д образования сгу стка (CFT), максимал ь ная п лот- ность сгустка (MCF)) больных онкологического п рофиля близки к контроль ным . CT у больных онкологического п рофиля составлял: у женщин — 50 с (38,3–58,7), у му жчин 50 (42–71,2) vs 51 (42–59), p = 0,1210/53 ( 42–74,8 ), p = 0,1975 в контрольной группе . CFT у таких пациентов составлял : у женщин — 72 ( 32–92,4 м жчин – 80 с (50,2–128,7) vs 78 (62–100), p = 0,0128 80 (59–124,4), p = 0,9384 в контрол ьной группе . MCF у больных онкологического п составлял: у женщин — 70 мм (59,9–82,5), у мужчин — 63 мм (56–73,7) vs 69 мм (59–95,8), p = 0,9911 / 69 мм (53,6–90), p = 0,0135 в контрол ьной группе. У женщинпоказатели вязкости крови MaxVel были выше, чем у му жчин . Показатели MaxVel повышены у таких пациентов : у женщин — 19 мм/100 с (14,3–49,5) у му жчин — 18 мм/100 (11–27 ) vs 15 мм / 100 (11,8–22), p < 0,001 / 13 мм / 100 с (10–21,8), p <0,001 в контрол ьной группе. ь t-MaxVel понижен у больных онкологического профиля: у женщин – 65 с (48,6–112,8) , у мужчин – 81 с (50,1–135,9) vs 115 с (56,8–166), p < 0,001 / 115 с (59,8–180,8), p = 0,0002 в контрольной группе. Показатель AUC у повышен у женщин — 6451 мм 100 (5511–8148), у мужчин — 5984 мм 100 (5119–6899) vs 5778 мм 100 (4998–6655), p < 0,001 / 5662 мм 100 (4704–6385), p = 0.0105. Выводы в отличие от других мето дов, измеря ющих вариации отдельных комп онентов системы крови, метод тромбо эластографии отражает текущийп рофиль образования сгу сткав режиме реаль ного времени является информативным споссобом оценки состояния гемостаза у онкологических больных. 2007 Article Thromboelastographic profiles as a tool for thrombotic risk in digestive tract cancer / M.L. Papa, F. Capasso, L. Pudore, S. Torre, S. Mango, V. Russo, P. Delrio, R. Palaia, F. Ruffolo, M.D. d’Eufemia, D. De Lucia, M. Napolitano, P. Di Micco, V. Parisi // Experimental Oncology. — 2007. — Т. 29, № 2. — С. 111-115. — Бібліогр.: 26 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/138581 en Experimental Oncology Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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Original contributions Original contributions Papa, M.L. Capasso, F. Pudore, L. Torre, S. Mango, S. Russo, V. Delrio, P. Palaia, R. Ruffolo, F. d’Eufemia, M.D. De Lucia, D. Napolitano, M. Di Micco, P. Parisi, V. Thromboelastographic profiles as a tool for thrombotic risk in digestive tract cancer Experimental Oncology |
description |
Background: Quantification of the magnitude of thrombotic risk associated with malignancy and with anti-cancer therapy is indispensable to use anticoagulant drugs which selectively interfere with haemostatic mechanisms protecting patients from venous thromboembolism (VTE) and probably from tumor progression. However, none of activation coagulation markers has any predictive value for the occurrence of the thrombotic events in one individual patient. Current clotting methods can’t reveal the overall dynamic clot formation; in contrast thromboelastographic methods specifically assess overall coagulation kinetics and its strength in whole blood. Aim: Objective of study was to evaluate if the activation of coagulation as eventually revealed by ROTEM® thromboelastometry could assess an hypercoagulable state in surgical neoplastic patients. Patients and Methods: Fifty consecutive patients with carcinoma of the digestive tract in preoperative period (23 M, 27 F aging 61.5 (45–79 years) and 147 healthy subjects (71 M, 76 F) were studied. A recent thromboelastometric method based on thrombelastography after Hartert was employed. Measurements were performed on ROTEM Coagulation Analyzer. The continuous coagulation data from 50 min course were transformed into dynamic velocity profiles of WB clot formation. Results: Standard parameters (CT, CFT, MCF) of cancer patients were similar to controls. CT (in cancer patients): females 50 s (38.3–58.7), males 50 s (42–71.2) vs 51 s (42–59), p = 0.1210 / 53 s (42–74.8), p = 0.1975 (in controls). CFT (in cancer patients): females 72 s (32- 92.4), males 80 s (50.2- 128.7) vs 78 s (62–100), p = 0.0128 / 80 s (59–124.4), p = 0.9384 (in controls). MCF (in cancer patients): females 70 mm (59.9–82.5), males 63 mm (56–73.7) vs 69 mm (59–95.8), p = 0.9911 / 69 mm (53.6–90), p = 0.0135 (in controls). Females showed a higher MaxVel when compared to males. The MaxVel was increased in cancer patients: females 19 mm /100 s (14.3–49.5) males 18 mm / 100 s (11–27) vs 15 mm 100 s (11.8–22), p < 0.001 / 13 mm / 100 s (10–21.8), p < 0.001 in controls .The t-MaxVel was shortened in cancer patients: females 65 s (48.6–112.8), males 81 s (50.1–135.9) vs 115 s (56.8–166), p <0.001 / 115 s (59.8–180.8), p = 0.0002 in controls. The AUC was increased in cancer patients: females 6451 mm 100 (5511–8148), males 5984 mm 100 (5119-6899) vs 5778 mm 100 (4998–6655), p < 0.001 / 5662 mm 100 (4704–6385), p = 0.0105. Conclusion: Unlike other assays measuring variations in a single component during coagulation, the thrombelastographic method records a profile of real-time continuous WB clot formation, and may provide extensive informations on haemostasis in neoplastic patients before surgery. |
format |
Article |
author |
Papa, M.L. Capasso, F. Pudore, L. Torre, S. Mango, S. Russo, V. Delrio, P. Palaia, R. Ruffolo, F. d’Eufemia, M.D. De Lucia, D. Napolitano, M. Di Micco, P. Parisi, V. |
author_facet |
Papa, M.L. Capasso, F. Pudore, L. Torre, S. Mango, S. Russo, V. Delrio, P. Palaia, R. Ruffolo, F. d’Eufemia, M.D. De Lucia, D. Napolitano, M. Di Micco, P. Parisi, V. |
author_sort |
Papa, M.L. |
title |
Thromboelastographic profiles as a tool for thrombotic risk in digestive tract cancer |
title_short |
Thromboelastographic profiles as a tool for thrombotic risk in digestive tract cancer |
title_full |
Thromboelastographic profiles as a tool for thrombotic risk in digestive tract cancer |
title_fullStr |
Thromboelastographic profiles as a tool for thrombotic risk in digestive tract cancer |
title_full_unstemmed |
Thromboelastographic profiles as a tool for thrombotic risk in digestive tract cancer |
title_sort |
thromboelastographic profiles as a tool for thrombotic risk in digestive tract cancer |
publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
publishDate |
2007 |
topic_facet |
Original contributions |
url |
http://dspace.nbuv.gov.ua/handle/123456789/138581 |
citation_txt |
Thromboelastographic profiles as a tool for thrombotic risk in digestive tract cancer / M.L. Papa, F. Capasso, L. Pudore, S. Torre, S. Mango, V. Russo, P. Delrio, R. Palaia, F. Ruffolo, M.D. d’Eufemia, D. De Lucia, M. Napolitano, P. Di Micco, V. Parisi // Experimental Oncology. — 2007. — Т. 29, № 2. — С. 111-115. — Бібліогр.: 26 назв. — англ. |
series |
Experimental Oncology |
work_keys_str_mv |
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2025-07-10T06:06:33Z |
last_indexed |
2025-07-10T06:06:33Z |
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fulltext |
Experimental Oncology ���� ��������� ����� ���ne�� ������� ��������� ����� ���ne�� �����ne�� ����� ��� ���
Veno�s thromboembolism �VTE�� is seven fold in-
creased in patients with malignancy when compared to
patients not affected by solid t�mors [�]. Malignancy
ind�ces an acq�ired thrombophilic state [�]; therefore��
in almost all cancer patients a s�b clinical activation of
blood coag�lation takes place�� even witho�t symptoms
of thrombosis. The hypercoag�lable state in cancer
arises mostly from the capacity of t�mo�r cells to ex-
press and release specific procoag�lant activities like
cancer procoag�lant �CPA�� and tiss�e factor �TF�� and
to interact with the host’s endotheli�m�� platelets and
monocytes-macrophages ind�cing a prothrombotic
phenotype within these cells [3]. The association be-
tween malignancy and VTE s�ch as the relationship
between t�mor growth and coag�lation activation has
been known since Tro�ssea�’s time [4]. Nowadays it is
evident that the coag�lation system plays an important
role in the biology of malignant t�mors: the activation
of haemostasis ind�ces a contin�o�s formation and
removal of fibrin which mediates the adhesion of t�mor
cells to endotheli�m facilitating their migration thro�gh
the tiss�es and contrib�ting to t�mor progression [�]. Re-
cent st�dies have revealed a nonhaemostatic role of TF
in generation of coag�lation proteases and s�bseq�ent
activation of proteases activated receptors �PARs�� on
vasc�lar cells. This TF dependent signaling contrib�tes
to a variety of biological processes incl�ding inflamma-
tion�� angiogenesis and metastasis [6]. The prothrombotic
state of cancer patients is enhanced by therape�tic in-
terventions�� s�ch as s�rgery�� chemotherapy�� hormone
therapy�� radiotherapy and it is related to disease stage
and to the site of origin of the primary t�mor: patients
with haematological cancer have the highest risk of VTE��
followed by those with l�ng and gastrointestinal cancers
[��� 6]. Therefore q�antification of the magnit�de of the
thrombotic risk associated with malignancy and anti-
cancer therapy is essential to �se anticoag�lant dr�gs
which selectively interfere with haemostatic mechanisms
probably protecting patients both from VTE and from
t�mor progression [��]. St�dies on haemostatic system in
cancer patients show an increase in clotting factors lev-
els�� markers of thrombin and fibrin generation�� fibrynolitic
proteins and thrombocytosis [8]. However�� none of the
coag�lation activation markers has any predictive val�e
ThromboelasTographic profiles as a Tool for
ThromboTic risk in digesTive TracT cancer
M.L. Papa1, F. Capasso1, L. Pudore1, S. Torre1, S. Mango1, V. Russo1, P. Delrio2, R. Palaia2, F. Ruffolo2,
M.D. d’Eufemia3, D. De Lucia3, *, M. Napolitano3, P. Di Micco4, V. Parisi2
1Laboratory of Haemostasis and Thrombosis, San Giovanni Bosco Hospital of Naples, Naples, Italy
2Division of Oncological Surgery, Istituto Nazionale Tumori, IRCCS, Fondazione Pascale, Naples, Italy
3Pathology Division, Second University of Naples, Naples, Italy
4Fatebenefratelli Hospital of Naples, Naples, Italy
Background: Quantification of the magnitude of thrombotic risk associated with malignancy and with anti-cancer therapy is indispensable
to use anticoagulant drugs which selectively interfere with haemostatic mechanisms protecting patients from venous thromboembolism
(VTE) and probably from tumor progression. However, none of activation coagulation markers has any predictive value for the occurrence
of the thrombotic events in one individual patient. Current clotting methods can’t reveal the overall dynamic clot formation; in contrast
thromboelastographic methods specifically assess overall coagulation kinetics and its strength in whole blood. Aim: Objective of study
was to evaluate if the activation of coagulation as eventually revealed by ROTEM® thromboelastometry could assess an hypercoagulable
state in surgical neoplastic patients. Patients and Methods: Fifty consecutive patients with carcinoma of the digestive tract in preoperative
period (23 M, 27 F aging 61.5 (45–79 years) and 147 healthy subjects (71 M, 76 F) were studied. A recent thromboelastometric method
based on thrombelastography after Hartert was employed. Measurements were performed on ROTEM Coagulation Analyzer. The con-
tinuous coagulation data from 50 min course were transformed into dynamic velocity profiles of WB clot formation. Results: Standard
parameters (CT, CFT, MCF) of cancer patients were similar to controls. CT (in cancer patients): females 50 s (38.3–58.7), males 50 s
(42–71.2) vs 51 s (42–59), p = 0.1210 / 53 s (42–74.8), p = 0.1975 (in controls). CFT (in cancer patients): females 72 s (32- 92.4),
males 80 s (50.2- 128.7) vs 78 s (62–100), p = 0.0128 / 80 s (59–124.4), p = 0.9384 (in controls). MCF (in cancer patients): females
70 mm (59.9–82.5), males 63 mm (56–73.7) vs 69 mm (59–95.8), p = 0.9911 / 69 mm (53.6–90), p = 0.0135 (in controls). Females
showed a higher MaxVel when compared to males. The MaxVel was increased in cancer patients: females 19 mm /100 s (14.3–49.5)
males 18 mm / 100 s (11–27) vs 15 mm 100 s (11.8–22), p < 0.001 / 13 mm / 100 s (10–21.8), p < 0.001 in controls .The t-MaxVel was
shortened in cancer patients: females 65 s (48.6–112.8), males 81 s (50.1–135.9) vs 115 s (56.8–166), p <0.001 / 115 s (59.8–180.8),
p = 0.0002 in controls. The AUC was increased in cancer patients: females 6451 mm 100 (5511–8148), males 5984 mm 100 (5119-
6899) vs 5778 mm 100 (4998–6655), p < 0.001 / 5662 mm 100 (4704–6385), p = 0.0105. Conclusion: Unlike other assays measuring
variations in a single component during coagulation, the thrombelastographic method records a profile of real-time continuous WB clot
formation, and may provide extensive informations on haemostasis in neoplastic patients before surgery.
Key Words: cancer, thromboelastography, hypercoagulable state, thromboelastometry, surgery.
Received: May 11, 2007.
*Correspondence: E-mail: domenico.delucia@unina2.it
Abbreviations used: ��C �� area under curve�� C��� �� clottin����C �� area under curve�� C��� �� clottin��C��� �� clottin��
formation time�� CP� �� cancer procoa��ulant�� C�� �� clottin�� time��
MC� �� maximum clot firmness�� P�Rs �� proteases activated re-
ceptors�� ��� �� tissue factor�� V��E �� venous thromboembolism.
Exp Oncol �����
���� ��� �������
��� Experimental Oncology ���� ��������� ����� ���ne��
for the occ�rrence of the thrombotic events in one in-
divid�al patient [�]. Since reliable methods�� that co�ld
have a higher predictive val�e for thrombosis risk�� are
needed�� o�r aim is to get better informations eval�ating
the profile of extended time coag�lation analysis. Th�s��
the investigation of coag�lation dynamics in whole blood
co�ld�� in o�r opinion�� disclose an abnormal pattern in
cancer patients�� especially in those at increased throm-
botic risk. C�rrent laboratory clotting techniq�es cannot
f�lly identify s�bjects with an increased thromboembolic
risk: their performance in plasma and the addition of
b�ffered sol�tions limit their relevance to overall dynamic
clot formation in whole blood [��]. In contrast ROTEM
thrombelastometry specifically assess overall coag�la-
tion kinetics and strength in whole blood�� providing a
global assessment of haemostatic f�nction [��].
Therefore the aim of this st�dy was to investi-
gate whether an hypercoag�lable state�� revealed by
ROTEM�� can be an important variable to eval�ate
the coag�lation derangements in patients affected by
cancer of the digestive tract.
maTerials and meThods
Patients. �� patients with histologically confirmed
solid cancer of the digestive tract were st�died.
�3 were male�� ��� were female and all aging 6�.� �4��
��� years��. The criteria for incl�sion in the st�dy have
been: patients with a carcinoma of the digestive tract
witho�t history of VTE who were candidate for s�rgery.
�4�� healthy s�bjects ���� M�� ��6 F�� selected on the basis
of sex and age were enrolled in o�r st�dy as control
gro�p. The st�dy was approved by Ethics Committee
of the Instit�te. Informed consent was collected from
all participants.
Sample collection. Blood samples were drawn be-
tween 8 am and � am�� after �� h of fasting. The blood
�nine vol�mes�� was placed in t�bes with �.��� M of
trisodi�m citrate �one vol�me�� �ntil analysis.
Test procedure. A recent thromboelastometric
method �ROTEM®; Pentapharm Ltd�� M�nich�� Ger-
many; distrib�ted in Italy by Dasit�� Milano���� based on
the thrombelastography after Hartert��was employed
[��]. Meas�rements were performed on a ROTEM
Coag�lation Analyzer. Citrated blood samples were
recalcified with CaCl� �star-TEM®�� reagent�� and ac-
tivated with tiss�e thromboplastin from rabbit brain
�ex-TEM�� reagent�� for monitoring the extrinsic system.
The ROTEM® analysis determines the onset of the
coag�lation process: clotting time �CT��; the kinetics of
clotting formation and stability: clotting formation time
�CFT�� and maxim�m clot firmness �MCF�� �Fig. ��� a��.
The ROTEM® software also calc�lates the novel pa-
rameters according to Sorensen [�4] s�ch as MaxVel��
t-MaxVel�� AUC by data derived from the ROTEM c�rve.
MaxVel describes the maxim�m velocity of the clot
formation. It is the maxim�m rate of clot formation�� the
maxim�m of the one derivative c�rve.
t-MaxVel describes the time from start of the mea-
s�rement till MaxVel �maxim�m of the one derivative��
is reached. It is a parameter similar to CT.
AUC describes the area �nder the velocity �one
derivative�� c�rve and is eq�ivalent to MCF in a c�rve
where the test had been r�n till MCF had been reached
[�4] �Fig. ��� b��.
fig. 1, a. Standard thrombelastographic tracking �CT�� CFT�� MCF��
fig. 1, b. ROTEM c�rve displayed by ROTEM Gamma software
Velocity profile�� the first derivative of the ROTEM tracking: maxi-
m�m velocity �MaxVel���� time to maxim�m velocity �t-MaxVel����
area �nder c�rve �AUC�� from Sorensen [�3]
Statistical analysis. Statistical significance of the
differences of val�es between patients and the healthy
controls was calc�lated by the Mann — Whitney U test.
The differences were considered statistically signifi-
cant only for p-val�es less �.�� �Tables ��� ���.
Table 1. RO��EM standard parameters in neoplastic patients and in
healthy reference subjects
Healthy
Males Median 5P 95P p value Patients
Males Median 5P 95P
C�� 53 42 74,8 0.1975 C�� 50 42 71,2
C��� 80 59 122,4 0.9384 C��� 80 50,2 128,7
MC� 69 53,6 90 0.0135 MC� 63 56 73,7
Healthy
�emales
Patients
�emales
C�� 51 42 59 0.1210 C�� 50 38,3 58,7
C��� 78 62 100 0.0128 C��� 72 32 92,4
MC� 69 59 95,8 0.9911 MC� 70 59,9 82,5
Medians and reference ran��e (5��95% percentile ) for controls and pa-
tients.
Table 2. RO��EM velocity parameters in neoplastic patients and in healthy
reference subjects
Healthy
Males Median 5P 95P p value Patient
Males Median P5 P95
MaxV 13 10 21,8 0.001 MaxV 18 11 27
MaxV-t 115 59.8 180.8 0.0002 MaxV-t 81 50.1 135.9
��C 5662 4703.6 6385 0.0105 ��C 5984 5118.7 6899.2
Healthy
�emales
Patients
�emales
MaxV 15 11.8 22 0.001 MaxV 19 14.3 49.5
MaxV-t 115 56.8 166 0.001 MaxV-t 65 48.6 112.8
��C 5778 4998 6655 0.001 ��C 6451 5514.8 8148.4
Medians and reference ran��e (5��95% percentile) for controls and for patients.
Experimental Oncology ���� ��������� ����� ���ne�� ��3���� ��������� ����� ���ne�� ��3��ne�� ��3�� ��3 ��3
resulTs
Standard parameters of ROTEM �CT�� CFT�� MCF�� were
not different in cancer patients as compared to controls.
CT �in cancer patients��: females �� s �38.3��8.������ males
�� s �4�����.��� vs �� s �4�������� p = �.����/�3 s �4����4.8����
p = �.����� �in controls��. CFT �in cancer patients��: females
��� s �3����.4���� males 8� s ���.����8.���� vs ��8 s �6���������
p = �.���8 / 8� s ������4.4���� p = �.�384 �in controls��.
MCF �in cancer patients��: females ��� mm ���.��8�.�����
males 63 mm ��6���3.���� vs 6� mm ������.8���� p = �.����/
6� mm ��3.6������� p = �.��3� �in controls��. The analysis
showed: an increase in MCF in male controls as compared
to patients and a shorter CFT in female patients than in
controls �Table ���. The contin�o�s coag�lation data from
a �� min-time co�rse were transformed into dynamic
velocity profiles of WB clot formation [�4] �Fig. ���.
fig. 2, a, b, c, d, e, f. �box plot��: medians and reference range
�����% percentile�� of MaxVel�� t-Max Vel�� AUC for controls and
patients
There were higher MaxVel in female controls
as compared to male controls. The res�lts were in
accordance with earlier st�dies [��]. The ROTEM®
velocity parameters res�lted significantly different in
patients when compared to the parameters of healthy
s�bjects. The MaxVel was increased �cancer patients��:
females �� mm ���/ s ��4.3�4�.����� males �8 mm
���/ s ��������� vs �� mm ���/s ���.8������� p <�.���/�3
mm ���/s ������.8���� p < �.��� �controls��. The t-
MaxVel was shortened �cancer patients��: females
6� s �48.6� ���.8���� males 8� s ���.���3�.��� vs ��� s
��6.8��66���� p < �.���/��� s ���.8��8�.8���� p = �.����
�controls��. The AUC was increased �cancer patients��:
females 64�� mm ��� ������8�48���� males ��84 mm
��� ������68���� vs �����8 mm ��� �4��8�66������ p <
�.���/�66� mm ��� �4���4�638����� p = �.���� �con-
trols�� �Table ���.
discussion
Reliable markers and methods to predict thrombo-
tic risk are essential to clinical management�� especially
for high- risk patients�� i. e.�� cancer patients �ndergoing
therape�tic interventions. Most laboratory tests act�-
ally �sed for st�dying haemostasis are performed on
platelet-poor plasma with clotting [��] or chromogenic
[�6] end points. Nowadays�� thanks to a better �nder-
standing of the role of platelets�� le�kocytes and eryth-
rocytes in the clotting process�� to eval�ate thrombin
generation d�ring blood coag�lation has become the
best approach in order to assess the global complex
process [���]. However�� whereas thrombin generation
tests are diffic�lt to perform in real time clinical prac-
tice [�8]�� thrombelastographic recording of the whole
blood coag�lation process is anticipated to indirectly
reflect the co�rse of thrombin generation .The classical
thrombelastography prod�ces a profile of the overall
rheological changes occ�rring d�ring coag�lation
and in the past it has been prevalently �sed to assist
clinicians in the control of after-s�rgery bleeding.
A newer modification of classical thrombelastography
is thromboelastometry �ROTEM®���� which avoids some
technical limitations of the traditional method�� s�ch as
sensitivity to vibrations or mechanical shocks. ROTEM
�ses a ball bearing system for power transd�ction
which makes it easily transportable and less s�scep-
tible to mechanical stress�� movement and vibration.
F�rthermore�� the activation of the samples accelerates
the meas�rement process and enhances reprod�cibil-
ity compared with conventional thromboelastography
[��]. The data obtained whit this new techniq�e are
contin�o�s�� digital and retrievable for f�rther calc�la-
tions; by processing of data the thrombelastographic
time co�rse can be transformed into a dynamic veloc-
ity profile of the changes in blood elasticity occ�rring
d�ring WB clot formation. The instr�ments software is
�sed to calc�late three new parameters in the assess-
ment of coag�lation dynamic properties. The pattern
of the new val�es: MaxVel�� t-MaxVel�� AUC�� display a
remarkable degree of similarity between endogeno�s
thrombin potential �thrombogram�� and thrombelasto-
graphic model. Therefore the profile of whole blood
coag�lation by thrombelastography�� as an indirect
meas�re of thrombin generation�� may provide exten-
sive informations on haemostasis�� not only for the clini-
cal management of bleeding b�t also for thrombophilic
states [��]. In vivo markers of coag�lation activation
�prothrombin fragment � + � / F� + ��� and fibrinolysis
�tiss�e plasminogen activator /t-PA�� correlate very well
��4 Experimental Oncology ���� ��������� ����� ���ne��
with ROTEM® clotting time �CT�� and maximal lysis �ML��
in a validation of rotation thrombelastography model
of systemic activation of coag�lation and fibrinolysis
[��]. TEG has been s�ccessf�lly �sed in clinical setting
to detect hypercoag�lable states. A postoperative hy-
percoag�lable state�� as revealed by TEG�� was associa-
ted with thrombotic complications in a wide gro�p of
s�rgical patients followed d�ring postoperative period
�ntil discharge [�3].
fig. 3. Velocity profiles of WB clot formation before s�rgery in neo-
plastic patients and in healthy s�bjects. �a�� Healthy male s�bject
�red�� and male patient �green�� velocity profile �b��. Healthy female
s�bject �red�� and female patient �green�� velocity profile
The correlation between thrombotic complications
and hypercoag�lability confirms that s�rgical patients
are at high risk for hypercoag�lability and that this plays
an important role in the pathogenesis of thrombosis. In
patients affected by malignant disease�� thrombosis is
the most freq�ent complication and the second ca�se
of death [�4]. S�rgical interventions in these patients
increase the risk of postoperative VTE �approximately
two-three fold�� in comparison to the risk in non-can-
cer patients �ndergoing the same proced�res. The
American College of Chest Physicians �ACCP�� has
stratified patients with malignancy in the highest risk
category of s�rgical patients and �rged ro�tine throm-
boprophylaxis [��]. Therefore�� especially for high-risk
patients�� it is strongly desirable to �se a one’s disposal
test characterized by a higher predictive val�e of the
thrombotic event.
Thrombelastography represents a val�able method
which monitors haemostasis �nder low shear environ-
ment as a whole dynamic process instead of revealing
information on isolated parts of the different linked path-
ways. Indeed thromboelastometry provides information
abo�t the whole process of clot formation which res�lts
from interdependent steps: coag�lation activation��
thrombin prod�ction�� fibrin formation and polymeriza-
tion�� platelet activation�� platelet-fibrin interaction.
In concl�sion�� the �se of thrombelastometric
method has red�ced the need of s�bstit�tive therapies
for the clinical management of bleeding problems d�r-
ing major s�rgical interventions �liver transplantations��
cardiovasc�lar proced�res�� ne�ros�rgery�� [�6]. Final-
ly�� the a�thors feel that the diagnosis of hypercoag�-
lable state by �sing thromboelastometry�� in patients
at higher thrombotic risk �especially cancer patients
d�ring s�rgery���� co�ld provide a rationale for more
targeted prophylactic antithrombotic treatments.
acknowledgemenTs
This st�dy was f�nded with grants in aid from DASIT
S. p. A Milno. Italy. We thank Dr. M. Lakner �Pentapharm
GmbH�� Germany�� and Dr. V. Scala �Dasit S. p. A�� Italy��
for their contrib�tions to the st�dy. The a�thors thankThe a�thors thank
also Dr Danilo Petraccaro on behalf of PC FIREWIRE
Society�� Naples�� Italy�� �e-mail: pcfirewire@j�mpy.it�� for
his s�pport on technical and graphic works.
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ТромбоэласТография как меТод оценки риска
Тромбоза при опухолях пищевариТельного ТракТа
Предпосылки исследования:: ��������������� ������ ����� ��������, ����������� �� ���������������� �������������� ���������������� ������ ����� ��������, ����������� �� ���������������� �������������� �
п�������пух�����й ����п��й, ���������ь�� ���ю���� � ����� п��������� ���д���-��������у�������, ��щ�щ�ющ�х ���ь����
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Цель: �п��д����ь, � ����й ���� ���������ь ����у������, �п��д�������й ����д�� ������э�����������, �������� ����������э�����������, �������� ���������������������, �������� ����������
��п���������������� ����� у ���ь��х ��������������� п������� п���� х��у���������� ���ш����ь����. Пациенты и мето�ето�
ды: �����д����� 50 ���ь��х ����� п�щ��������ь���� ������ � д��п���������й п����д (27 ���щ��, 23 �у�����, ���д��й����� п�щ��������ь���� ������ � д��п���������й п����д (27 ���щ��, 23 �у�����, ���д��й п�щ��������ь���� ������ � д��п���������й п����д (27 ���щ��, 23 �у�����, ���д��й
������� 61,5 ��д� (45–79 ���) � 147 �д�����х д������ (71 �у�����, 76 ���щ��). ���������� ����д ������э�����������,. ���������� ����д ������э�����������,
���������й �� ������э����������� Г�������, � ��п��ь�������� ����������� ����у������ ����� ROTEM. ���ущ�� д�����ROTEM. ���ущ�� д����� ���ущ�� д�����
� ����������� �� 50 ��� ��������й п��д������� � ��д� д��������х п������й ��������� п�� ����������� ��у���� �����.
Результаты: ����д������ п�������� (п����д ����у������ (CT), п����д ������������ ��у���� (CFT), ��������ь���� п���-����д������ п�������� (п����д ����у������ (CT), п����д ������������ ��у���� (CFT), ��������ь���� п���- (п����д ����у������ (CT), п����д ������������ ��у���� (CFT), ��������ь���� п���-п����д ����у������ (CT), п����д ������������ ��у���� (CFT), ��������ь���� п���-CT), п����д ������������ ��у���� (CFT), ��������ь���� п���-), п����д ������������ ��у���� (CFT), ��������ь���� п���-, п����д ������������ ��у���� (CFT), ��������ь���� п���-п����д ������������ ��у���� (CFT), ��������ь���� п���-CFT), ��������ь���� п���-), ��������ь���� п���-, ��������ь���� п���-��������ь���� п���-
����ь ��у���� (MCF)) ���ь��х ��������������� п������� ������ � �������ь���. CT у ���ь��х ��������������� п�������MCF)) ���ь��х ��������������� п������� ������ � �������ь���. CT у ���ь��х ��������������� п�������) ���ь��х ��������������� п������� ������ � �������ь���. CT у ���ь��х ��������������� п������� ���ь��х ��������������� п������� ������ � �������ь���. CT у ���ь��х ��������������� п����������ь��х ��������������� п������� ������ � �������ь���. CT у ���ь��х ��������������� п�������. CT у ���ь��х ��������������� п�������у ���ь��х ��������������� п������� ���ь��х ��������������� п����������ь��х ��������������� п�������
����������: у ���щ�� �� 50 � (38,3–58,7), у �у���� �� 50 � (42–71,2) vs 51 � (42–59),: у ���щ�� �� 50 � (38,3–58,7), у �у���� �� 50 � (42–71,2) vs 51 � (42–59),у ���щ�� �� 50 � (38,3–58,7), у �у���� �� 50 � (42–71,2) vs 51 � (42–59), 50 � (38,3–58,7), у �у���� �� 50 � (42–71,2) vs 51 � (42–59),� (38,3–58,7), у �у���� �� 50 � (42–71,2) vs 51 � (42–59), (38,3–58,7), у �у���� �� 50 � (42–71,2) vs 51 � (42–59),у �у���� �� 50 � (42–71,2) vs 51 � (42–59), �� 50 � (42–71,2) vs 51 � (42–59),�� 50 � (42–71,2) vs 51 � (42–59),50 � (42–71,2) vs 51 � (42–59),� (42–71,2) vs 51 � (42–59), (42–71,2) vs 51 � (42–59),� (42–59), (42–59), p = 0,1210/53 � ( 42–74,8 ),0,1210/53 � ( 42–74,8 ),� ( 42–74,8 ), ( 42–74,8 ), p = 0,1975 0,1975
� �������ь��й ��упп�. CFT у ����х п�������� ����������: у ���щ�� �� 72 � ( 32–92,4) у �у���� – 80 � (50,2–128,7) vs. CFT у ����х п�������� ����������: у ���щ�� �� 72 � ( 32–92,4) у �у���� – 80 � (50,2–128,7) vsу ����х п�������� ����������: у ���щ�� �� 72 � ( 32–92,4) у �у���� – 80 � (50,2–128,7) vs ����х п�������� ����������: у ���щ�� �� 72 � ( 32–92,4) у �у���� – 80 � (50,2–128,7) vs����х п�������� ����������: у ���щ�� �� 72 � ( 32–92,4) у �у���� – 80 � (50,2–128,7) vs п�������� ����������: у ���щ�� �� 72 � ( 32–92,4) у �у���� – 80 � (50,2–128,7) vs����������: у ���щ�� �� 72 � ( 32–92,4) у �у���� – 80 � (50,2–128,7) vs: у ���щ�� �� 72 � ( 32–92,4) у �у���� – 80 � (50,2–128,7) vsу ���щ�� �� 72 � ( 32–92,4) у �у���� – 80 � (50,2–128,7) vs �� 72 � ( 32–92,4) у �у���� – 80 � (50,2–128,7) vs�� 72 � ( 32–92,4) у �у���� – 80 � (50,2–128,7) vs72 � ( 32–92,4) у �у���� – 80 � (50,2–128,7) vs� ( 32–92,4) у �у���� – 80 � (50,2–128,7) vs ( 32–92,4) у �у���� – 80 � (50,2–128,7) vs) у �у���� – 80 � (50,2–128,7) vs у �у���� – 80 � (50,2–128,7) vsу �у���� – 80 � (50,2–128,7) vs – 80 � (50,2–128,7) vs� (50,2–128,7) vs(50,2–128,7) vs
78 � (62–100), p = 0,0128 / 80 � (59–124,4),� (62–100), p = 0,0128 / 80 � (59–124,4), (62–100), p = 0,0128 / 80 � (59–124,4), = 0,0128 / 80 � (59–124,4), 0,0128 / 80 � (59–124,4), / 80 � (59–124,4),/ 80 � (59–124,4), 80 � (59–124,4),80 � (59–124,4),� (59–124,4), (59–124,4), p = 0,9384 � �������ь��й ��упп�. MCF у ���ь��х ��������������� п������� 0,9384 � �������ь��й ��упп�. MCF у ���ь��х ��������������� п�������� �������ь��й ��упп�. MCF у ���ь��х ��������������� п�������. MCF у ���ь��х ��������������� п�������у ���ь��х ��������������� п������� ���ь��х ��������������� п����������ь��х ��������������� п������� ��������������� п���������������������� п�������
����������: у ���щ�� �� 70 �� (59,9–82,5), у �у���� �� 63 �� (56–73,7) vs 69 �� (59–95,8),: у ���щ�� �� 70 �� (59,9–82,5), у �у���� �� 63 �� (56–73,7) vs 69 �� (59–95,8),у ���щ�� �� 70 �� (59,9–82,5), у �у���� �� 63 �� (56–73,7) vs 69 �� (59–95,8), �� 70 �� (59,9–82,5), у �у���� �� 63 �� (56–73,7) vs 69 �� (59–95,8),�� 70 �� (59,9–82,5), у �у���� �� 63 �� (56–73,7) vs 69 �� (59–95,8),70 �� (59,9–82,5), у �у���� �� 63 �� (56–73,7) vs 69 �� (59–95,8),�� (59,9–82,5), у �у���� �� 63 �� (56–73,7) vs 69 �� (59–95,8), (59,9–82,5), у �у���� �� 63 �� (56–73,7) vs 69 �� (59–95,8),у �у���� �� 63 �� (56–73,7) vs 69 �� (59–95,8), �� 63 �� (56–73,7) vs 69 �� (59–95,8),�� 63 �� (56–73,7) vs 69 �� (59–95,8),63 �� (56–73,7) vs 69 �� (59–95,8),�� (56–73,7) vs 69 �� (59–95,8), (56–73,7) vs 69 �� (59–95,8),�� (59–95,8), (59–95,8), p = 0,9911 / 69 �� (53,6–90), 0,9911 / 69 �� (53,6–90),�� (53,6–90), (53,6–90),
p = 0,0135 � �������ь��й ��упп�. �� ���щ�� п��������� ��������� ����� MaxVel ���� ��ш�, ��� у �у����. ���������� 0,0135 � �������ь��й ��упп�. �� ���щ�� п��������� ��������� ����� MaxVel ���� ��ш�, ��� у �у����. ����������,0135 � �������ь��й ��упп�. �� ���щ�� п��������� ��������� ����� MaxVel ���� ��ш�, ��� у �у����. ����������0135 � �������ь��й ��упп�. �� ���щ�� п��������� ��������� ����� MaxVel ���� ��ш�, ��� у �у����. ����������� �������ь��й ��упп�. �� ���щ�� п��������� ��������� ����� MaxVel ���� ��ш�, ��� у �у����. ����������. �� ���щ�� п��������� ��������� ����� MaxVel ���� ��ш�, ��� у �у����. ������������ ���щ�� п��������� ��������� ����� MaxVel ���� ��ш�, ��� у �у����. ���������� п��������� ��������� ����� MaxVel ���� ��ш�, ��� у �у����. ����������п��������� ��������� ����� MaxVel ���� ��ш�, ��� у �у����. ���������� ��������� ����� MaxVel ���� ��ш�, ��� у �у����. ������������������� ����� MaxVel ���� ��ш�, ��� у �у����. ����������MaxVel ���� ��ш�, ��� у �у����. �������������� ��ш�, ��� у �у����. ����������. ��������������������
MaxVel п���ш��� у ����х п��������: у ���щ�� �� 19 ��/100 � (14,3–49,5) у �у���� �� 18 ��/100 � (11–27 ) vs 15 �� /п���ш��� у ����х п��������: у ���щ�� �� 19 ��/100 � (14,3–49,5) у �у���� �� 18 ��/100 � (11–27 ) vs 15 �� /: у ���щ�� �� 19 ��/100 � (14,3–49,5) у �у���� �� 18 ��/100 � (11–27 ) vs 15 �� /у ���щ�� �� 19 ��/100 � (14,3–49,5) у �у���� �� 18 ��/100 � (11–27 ) vs 15 �� / �� 19 ��/100 � (14,3–49,5) у �у���� �� 18 ��/100 � (11–27 ) vs 15 �� /�� 19 ��/100 � (14,3–49,5) у �у���� �� 18 ��/100 � (11–27 ) vs 15 �� /19 ��/100 � (14,3–49,5) у �у���� �� 18 ��/100 � (11–27 ) vs 15 �� /��/100 � (14,3–49,5) у �у���� �� 18 ��/100 � (11–27 ) vs 15 �� /100 � (14,3–49,5) у �у���� �� 18 ��/100 � (11–27 ) vs 15 �� /� (14,3–49,5) у �у���� �� 18 ��/100 � (11–27 ) vs 15 �� / (14,3–49,5) у �у���� �� 18 ��/100 � (11–27 ) vs 15 �� /у �у���� �� 18 ��/100 � (11–27 ) vs 15 �� / �� 18 ��/100 � (11–27 ) vs 15 �� /�� 18 ��/100 � (11–27 ) vs 15 �� /18 ��/100 � (11–27 ) vs 15 �� /��/100 � (11–27 ) vs 15 �� /100 � (11–27 ) vs 15 �� /� (11–27 ) vs 15 �� / (11–27 ) vs 15 �� /�� /
100 � (11,8–22),� (11,8–22), (11,8–22), p < 0,001 / 13 �� / 100 � (10–21,8),�� / 100 � (10–21,8), / 100 � (10–21,8),/ 100 � (10–21,8),100 � (10–21,8), � (10–21,8), (10–21,8), p <0,001 � �������ь��й ��упп�. ���������ь t-MaxVel п������ у� �������ь��й ��упп�. ���������ь t-MaxVel п������ у. ���������ь t-MaxVel п������ у ���������ь t-MaxVel п������ уt-MaxVel п������ у-MaxVel п������ уMaxVel п������ у п������ у
���ь��х ��������������� п�������: у ���щ�� – 65 � (48,6–112,8) , у �у���� – 81 � (50,1–135,9) vs 115 � (56,8–166),vs 115 � (56,8–166), 115 � (56,8–166),
p < 0,001 / 115 � (59,8–180,8), p = 0,0002 � �������ь��й ��упп�. ���������ь AUC у п���ш�� у ���щ�� �� 6451 �� 100AUC у п���ш�� у ���щ�� �� 6451 �� 100 у п���ш�� у ���щ�� �� 6451 �� 100
(5511–8148), у �у���� �� 5984 �� 100 (5119–6899) vs 5778 �� 100 (4998–6655),vs 5778 �� 100 (4998–6655), 5778 �� 100 (4998–6655), p < 0,001 / 5662 �� 100 (4704–6385),
p = 0.0105. Выводы:: � ������� �� д�у��х ����д��, �������ющ�х �������� ��д��ь��х ���п������� ������� ������������� ������� �� д�у��х ����д��, �������ющ�х �������� ��д��ь��х ���п������� ������� ������������ ������� �� д�у��х ����д��, �������ющ�х �������� ��д��ь��х ���п������� ������� ������������������� �� д�у��х ����д��, �������ющ�х �������� ��д��ь��х ���п������� ������� ������������ �� д�у��х ����д��, �������ющ�х �������� ��д��ь��х ���п������� ������� �������������� д�у��х ����д��, �������ющ�х �������� ��д��ь��х ���п������� ������� ������������ д�у��х ����д��, �������ющ�х �������� ��д��ь��х ���п������� ������� ������������д�у��х ����д��, �������ющ�х �������� ��д��ь��х ���п������� ������� ������������ ����д��, �������ющ�х �������� ��д��ь��х ���п������� ������� ����������������д��, �������ющ�х �������� ��д��ь��х ���п������� ������� ������������, �������ющ�х �������� ��д��ь��х ���п������� ������� �������������������ющ�х �������� ��д��ь��х ���п������� ������� ������������ �������� ��д��ь��х ���п������� ������� �������������������� ��д��ь��х ���п������� ������� ������������ ��д��ь��х ���п������� ������� ��������������д��ь��х ���п������� ������� ������������ ���п������� ������� ���������������п������� ������� ������������ ������� ������������������� ������������ ������������������������
�����, ����д ������э����������� �������� ���ущ�й п�����ь ������������ ��у���� � ������ ����ь���� ������� � �����������, ����д ������э����������� �������� ���ущ�й п�����ь ������������ ��у���� � ������ ����ь���� ������� � ���������������д ������э����������� �������� ���ущ�й п�����ь ������������ ��у���� � ������ ����ь���� ������� � ����������� ������э����������� �������� ���ущ�й п�����ь ������������ ��у���� � ������ ����ь���� ������� � �����������������э����������� �������� ���ущ�й п�����ь ������������ ��у���� � ������ ����ь���� ������� � ����������� �������� ���ущ�й п�����ь ������������ ��у���� � ������ ����ь���� ������� � ������������������� ���ущ�й п�����ь ������������ ��у���� � ������ ����ь���� ������� � ����������� ���ущ�й п�����ь ������������ ��у���� � ������ ����ь���� ������� � ��������������ущ�й п�����ь ������������ ��у���� � ������ ����ь���� ������� � ����������� п�����ь ������������ ��у���� � ������ ����ь���� ������� � �����������п�����ь ������������ ��у���� � ������ ����ь���� ������� � ����������� ������������ ��у���� � ������ ����ь���� ������� � ����������������������� ��у���� � ������ ����ь���� ������� � ����������� ��у���� � ������ ����ь���� ������� � �������������у���� � ������ ����ь���� ������� � ����������� � ������ ����ь���� ������� � ������������ ������ ����ь���� ������� � ����������� ������ ����ь���� ������� � ����������������� ����ь���� ������� � ����������� ����ь���� ������� � ���������������ь���� ������� � ����������� ������� � ������������������ � ����������� � ������������ ����������� ����������������������
������������� �п������� ������ ����������� ��������� у �������������х ���ь��х. �п������� ������ ����������� ��������� у �������������х ���ь��х.�п������� ������ ����������� ��������� у �������������х ���ь��х. ������ ����������� ��������� у �������������х ���ь��х.������ ����������� ��������� у �������������х ���ь��х. ����������� ��������� у �������������х ���ь��х.����������� ��������� у �������������х ���ь��х. ��������� у �������������х ���ь��х.��������� у �������������х ���ь��х. у �������������х ���ь��х.у �������������х ���ь��х. �������������х ���ь��х.�������������х ���ь��х. ���ь��х.���ь��х..
Ключевые слова: слова:слова:: ���, ������э������������, ���������� ��п������у������, ������э������������, х��у�����.���, ������э������������, ���������� ��п������у������, ������э������������, х��у�����., ������э������������, ���������� ��п������у������, ������э������������, х��у�����.������э������������, ���������� ��п������у������, ������э������������, х��у�����., ���������� ��п������у������, ������э������������, х��у�����.���������� ��п������у������, ������э������������, х��у�����. ��п������у������, ������э������������, х��у�����.��п������у������, ������э������������, х��у�����., ������э������������, х��у�����.������э������������, х��у�����..
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