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Journal of Health Sciences (J Health Sci) 2012- (2)6: 80−88 The journal has had 4 points in Ministry of Science and Higher Education of Poland parametric evaluation. List B item 683
AI Gozhenko1, AN Likhoded2, VV Shukhtin1, AV Bogdanova1, W Zukow3 А.И. Гоженко1, А.Н. Лиходед2, В. В. Шухтин1, А.В. Богданова1, В.А. Жуков3
1SE Ukrainian Scientific-Research Institute of Transport Medicine, Odessa, Ukraine 2KU Odessa Regional Medical Center, Odessa, Ukraine 3Radom University, Radom, Poland
1ГП Украинский научно-исследовательский институт медицины транспорта 2КУ «Одесский Областной Медицинский Центр»
3Radom University, Radom Poland
© The Author (s) 2012-
This article is published with open access at Licensee Open Journal Systems of Radom University in Radom, Poland
Keywords: pyelonephritis, urinary syndrome, inflammation, markers of CD38 and CD45. Ключевые слова: пиелонефрит, мочевой синдром, воспаление, маркеры CD38 и CD45.
35 persons of sufferings are inspected by a pyelonephritis. By a research purpose, there was a study of indexes of subpopulation of lymphocytes of urine by an immune complex peroxidase-antiperoxidase (markers of CD38 and CD45). The method of determination of lymphocytes of CD38 and CD45 is developed in urine, certain their amount at healthy persons. As a result of the conducted research it was exposed for patients a pyelonephritis increase of subpopulation of lymphocytes of CD38 and CD45 in urine.
Обследовано 35 человек страдающих пиелонефритом. Целью исследования, было изучение в моче показателей маркеров активации лейкоцитов (субпопуляции СD38 и СD45) с помощью иммунного комплекса пероксидаза-антипероксидаза. Разработана методика определения лейкоцитов CD38 и CD45 в моче, определено их количество у здоровых лиц. В результате проводимого исследования было выявлено у больных пиелонефритом увеличение субпопуляций лейкоцитов CD38 и CD45 в моче.
Relevance of the topic: pyelonephritis refers to inflammatory diseases, the etiology of which play a role as many microorganisms are constantly living in the human body, belonging to the normal human microflora, populate the skin and mucous membranes — endogenous flora and microorganisms living in the environment (exogenous flora), bacteria, fungi such as Candida, viruses, mycoplasma [3,7,12]. For diagnosis of inflammatory genes in the kidney, as well as their activity is most often study the qualitative changes Urine: bacteriuria, proteinuria, pyuria, cylindruria [1,2,5]. However, it is shown that white blood cells in the urine detected in healthy persons, and pyelonephritis of them, as a rule, considerably increases particularly acute process.
However, of particular interest is diagnosis of chronic pyelonephritis, in which the urine is not always revealed a large number of white blood cells. In order to improve the accuracy of diagnosis of inflammatory processes in the kidney, proposed the definition of active leukocytes in urine, called cells Shterngeymer — Malbin whose presence is especially true for chronic pyelonephritis [4,6]. Meanwhile it was established that in some cases with pyelonephritis, such as alkaline urine can not
identify leukocytes, and their conversion into cells Shterngeymera — Malbina more dependent on the reduction of urine osmolality than the presence of inflammation in the kidneys [9,10, 11]. In connection with the above, we have developed and tested a method definition in the urine leukocyte activation markers CD38 and CD45, which are increased in the blood is considered as evidence of inflammatory processes [13,14,15].
Research goal was to compare the diagnostic indicators of inflammation in the kidney (bladder syndrome) and the definition of leukocyte activation markers CD38, CD45.
Materials and Methods: We examined 10 healthy subjects and 17 patients suffering from pyelonephritis in age from 40 years to 68 years, the average age of patients was 55 years (the distribution of the patients were in 2 groups). Group comprised 7 patients pyelonephritis without comorbidity, the second group included 10 patients of whom had concomitant diseases (diabetes, hypertension), the average age of this group was 60 years.
A control group of 10 people who do not have inflammatory kidney disease, the average age of this group was — 45 years.
The clinical diagnosis of pyelonephritis established according the history, clinical and laboratory studies (increased creatinine, urine analysis data Nechipurenko urine on Zimnitskiy, also performed ultrasonography in patients to pay attention — the extension of renal pelvis, rough outlines of the cups, the heterogeneity of the parenchyma with areas scarring. Additionally the number of patients the radioisotope renography. in the urine along with the determination of the total number of cells was determined subset of leukocyte CD38, CD45 according to our methodology.
Determination of leukocyte subpopulations of cells with immune complex-peroxidase-antiperoxidase. For this 4 ml. mochi diluted with saline at a ratio of 1 to 2, and centrifuged for 15 min., At 1800 rpm., At room temperature. Supernatant was decanted, added saline, resuspended and centrifuged at 1000 rpm. min., at room
temperature. The washed leukocytes were diluted with saline, bringing them to a concentration of 4.2×10 cells / ml. On cytological preparations consistently applied 100 L monoclonal antibody CD38, SD45 and complex horseradish peroxidase-antiperoksidaza. Finished stained with methylene green and horseradish peroxidase activity was determined to identify the various lymphocyte subpopulations. Cells with detectable antigen linked to horseradish peroxidase, were along the cytoplasmic dark brown rim.
Results and discussion
In the control group there were no changes in urine specific to bladder syndrome. The number of leukocytes by Nechipurenko not exceed 2000, and the red blood cells were within 1000, also did not show proteinuria and bacteriuria. However, the white blood cells found in the urine marker CD38 index was in the range of up to 4%, and CD45 that does not exceed 8%.
However, in patients with pyelonephritis revealed the presence of urinary symptoms. Since all patients 1 and 2 groups found proteinuria, increased number of white blood cells and red blood cells (Table 1) for bacteriological examination of urine culture in the patients examined pathogens seeded in all cases, but the degree of bacteriuria ranged from 104 to 107 degrees. According to the results of bacteriological and microscopy studies determined the most E. coli, 75% to 80%, against 60%, 75%, 30%, Staphylococcus, Klebsiella, 15%, in all other cases — the microbial association of 5%, Pseudomonas Wand — 1% -2%, 0%, streptococci. In general urine analysis revealed more than 10 white blood cells per field of view.
Table 1. Urinary symptoms and figures leukocyturia patients with pyelonephritis.
Of indices Исследуемый показатель Protein, mg / ml Белок, мг/мл Leukocytes Лейкоциты Erythrocytes Эритроциты Leukocyte CD38 Лейкоциты CD38 Leukocytes CD45 Лейкоциты CD45
Sick Group 1 Without comorbidity Больные 1 группа Без сопутствующей патологии 0,104 30,000 2,100 16% 21%
0,104 10,000 2,300 28% 30%
0,104 10,500 3,500 16% 20%
0,099 15,000 3,200 12% 24%
0,104 8,750 2,000 14% 24%
0,096 50,000 2,500 18% 32%
0,096 20,000 2,500 16% 20%
Sick Group 2 With comorbidities Больные 2 группа С сопутствующей патологией 0,104 20,500 3,000 15% 24%
0,096 10,000 3,500 14% 24%
0,302 15,000 3,000 15% 24%
0,104 8,500 3,200 14% 20%
0,096 20,000 2,500 16% 32%
0,096 20,500 3,500 15% 24%
0,302 45,000 3,000 16% 32%
0,104 50,000 3,200 18% 32%
0,099 30,000 3,500 15% 20%
0,104 15,000 3,000 14% 20%
In patients with pyelonephritis in the urine by Nechipurenko found an increase in the number of white blood cells and white blood cells CD38, CD45 (Table 1). These data indicate that CD38 is increased in all patients 3 times or more, and the number of white blood cells and CD45 also increased to 32%. The indices CD38 and CD45 more than 2−3 times the value determined in healthy subjects. It should be noted that significant differences in terms of CD38 and CD45 were found between the patients of the first and second group. Also found no relationship between leukocyturia, proteinuria, red blood cell and the activity of white blood cells by CD38 and CD45.
Thus increasing leukocyte CD38, CD45 in urine is an important indicator of inflammation in the kidney. Given that according to the literature the same changes in the blood, tears testify activation of leukocytes, which is observed in the inflammatory process, it is believed that in this way you can judge the presence and even the stage of the inflammatory process in the kidneys. Moreover, these changes reflect the active immune response and inflammation caused by the change of physical and chemical composition of urine. Therefore the determination of lymphocyte subpopulations is prognostically more reliable for the diagnosis of inflammatory diseases of the kidney test than the definition of active cell Shterngeymera — Malbina and total white cell count urine and other indicators of bladder syndrome.
1. In the urine of patients with pyelonephritis defined leukocyte CD38 and CD45 in excess in healthy individuals.
2. In the diagnosis of inflammation in the kidneys with pyelonephritis can be used method of determination of leukocyte CD38 and CD45 with monoclonal antibodies.
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This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http: //creativecommons. org/licenses/by-nc/3. 0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.
Received: 10. 12. 2012.
Revised: 25. 12. 2012.
Accepted: 26. 12. 2012.

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