Method for assessing endometrial cancer progression in patients with hyperplastic processes in endometrium

FIELD: medicine.

SUBSTANCE: clinical anamnestic parameters are determined in the patients with hyperplastic processes in the endometrium, and a hysteroscopy with an endometrial biopsy is performed. The activity of calpains, total activity of proteasomes, and activity of 20S proteasomes are determined. Age-dependent discriminator functions Y1 and Y2 are determined by formulas. If Y1>Y2, the patient is referred to a group of low risk of endometrial cancer development. If Y1<Y2, the patient is referred to a group of high risk of endometrial cancer development.

EFFECT: invention enables the high-sensitivity and specificity prediction of endometrial cancer development in the patients suffering from the hyperplastic processes in the endometrium.

1 tbl, 3 ex

 

The invention relates to medicine, specifically to Oncology, ways of assessing the risk of developing endometrial cancer in patients with hyperplastic processes of the endometrium.

Endometrial cancer (EC) is one of the most common types of gynecologic cancer pathology, in recent decades there has been a significant rejuvenation of the patients. In this regard, re is an important health problem and is characterized by high costs of health care resources. Secondary prevention of re is an important way in reducing morbidity and mortality from PE. A promising direction in solving this problem, the objective is the formation of groups at increased cancer risk, especially among patients with hyperplastic processes of the endometrium (GOE).

Objective and reliable criteria to predict cancer risk in patients with ET in the known sources of information are few and far between. Attempts were made to develop algorithms for prediction of malignancy of benign tumors of the endometrium based on clinical and histological parameters, however, this approach proved to be effective only for patients with atypical hyperplasia. One of the existing methods of determining cancer risk among patients with Predrag�Ohm endometrium is the decision rule, built with consideration of clinical and morphological parameters (duration of menstrual cycle, the number of abortions, number of audiiton in the endometrium, the number of pathological mitoses, mitotic index, the activity of proto-oncogenes bcl-2) and classification of sick of atypical hyperplasia to the class "cancer" or "precancer" [6].

Another known method is based on the levels of estrone and progesterone in the serum of the patient, the level of ER in the endometrium, the activity of enzymes of estrogen metabolism and a number of clinical and anamnestic parameters (age at menarche, presence/absence of uterine fibroids and hypertension) [5]. Based on the analysis of clinical data, including patient age, endometrial thickness, frequency of bleeding, body mass index and the presence or absence of diabetes mellitus, N. Burbos et al. built model "Norwich DEFAB" to calculate the risk of developing endometrial carcinoma in patients with vaginal bleeding in menopause [9]. However, these approaches are of limited use, due either to the complexity of performance analysis, or with restrictions on age of the patient or the type of hyperplasia.

Known methods of risk assessment based on the determination of molecular parameters. For assessment of risk for PE in patients with atypical GOE proposed definition of the expression of membrane�x protein of syndecan-1, in patients with polyps markers of angiogenesis VEGFB, VEGFR2, CD31 and CD34 [12, 13].

The known method of determination of indications for hysterectomy in patients with endometrial hyperplasia according to the level of tumor markers VEGF and p53 protein in endometrial tissue: when more than 1 value of the first score and the second detection make the conclusion about the necessity of hysterectomy [3].

Closest to the claimed method is a method based on using a mathematical model, proposed by L. Hunanyan et al., which includes both clinical history and molecular indicators: type of the GOE, the hereditary history of cancer patients, other diseases of reproductive system, neuroendocrine disorders, the expression of Claudine-3, PTEN, Ki-67 [4].

The disadvantages of this method are:

- lack of sensitivity and specificity that makes it impossible to assess the practical significance and the efficacy of the method;

- limitations associated with the age of the patient;

- a large number of defined parameters.

New technical problem - increasing the accuracy of determining the possible outcome of the disease in relation to malignancy tissue.

To solve the problem in the method of assessing the risk of developing endometrial cancer in patients with hyperplastic processes of the endometrium, on�waiting for a hysteroscopy with endometrial biopsy and determination of clinical and anamnestic indicators in the endometrium define activity kalainov, total proteasome activity, the activity of the 20S proteasome, also take into account the patient's age, then calculate the discriminant functions Y1 and Y2 in the formulas:

Y1=0,0285×X1+0,0091×X2+0,5429×X3-0,0031×X4-14,6595;

Y2=0,0517×X1+0,0239×x2+0,6568×X3-0,0072×X4-21,0753, where

X1 - activity kalainov, ·103U/mg protein

X2 - total proteasome activity, ·103U/mg protein

X3 - the patient's age, years

X4 - 20S activity of a pool of proteasomes, ·103U/mg protein

and when Y1>Y2, the patient is referred to the group with low risk, and if Y1<Y2 - to the group with a high risk of developing endometrial cancer.

The method is as follows.

For assessment of risk for PE in patients with ET is carried out to determine the activity of kalainov, total activity of the proteasome, activity of 20S proteasomes in the endometrium. To do this during hysteroscopy with biopsy taking a tissue sample and placed in liquid nitrogen. The tissue samples to determine the activity of enzymes stored at -70°C and thawed no more than 1 times. Before determining the activity of the investigated enzymes frozen tumor tissue was homogenized to a powder in liquid nitrogen. 20S proteasomes isolated by salting out method using ammonium sulfate to 70% saturation [2]. Chymotrypsinogen proteasome activity and activity �of alpinev appreciate the hydrolysis fluorogenic of oligopeptides Suc-LLVY-AMC (Sigma, USA) [7]. The resulting product register on the fluorimeter "Hitachi-850 (Japan) at an excitation wavelength of 380 nm and emission 440 nm. To evaluate the activity of protease impurity is used, a specific proteasome inhibitor is MG132 and specific inhibitor of kalainov - ALLN. Per unit activity of proteasomes and kalainov take the amount of enzyme which hydrolyses 1 nmol Suc-LLVY-AMC for 1 min. Specific activity of proteasomes and kalainov expressed in units of activity per 1 mg of protein. The protein content was determined by Lowry's method. Also take into account the patient's age, then calculate the discriminant functions Y1 and Y2 in equations.

Y1=0,0285×X1+0,0091×x2+0,5429×X3-0,0031×X4-14,6595;

Y2=0,0517×X1+0,0239×x2+0,6568×X3-0,0072×X4-21,0753, where

where X1 is the activity of kalainov, ·103U/mg protein).

X2 - total proteasome activity, ·103U/mg protein).

X3 - the patient's age, years;

X4 - 20S activity of a pool of proteasomes, ·103U/mg protein.

and when Y1>Y2 the patient is referred to the group with a low probability, and if Y1<Y2 the patient is referred to the group with a high probability of developing endometrial cancer.

The proposed method is based on the analysis of clinical trial data. The approach to the assessment of risk of development of PE in patients with ET is due to a number of assumptions:

The unpredictability of the biological behavior of modified tissue and the possibility of its m�lignisul largely due to insufficient knowledge of the complex mechanisms of tumor growth and the lack of data on the extent of involvement in these processes of molecular and biochemical changes, occur in cells at different stages of carcinogenesis.

Know the importance of the work of the proteolytic systems in the development and progression of tumors [11]. So, for PE it shows the increased proteasome - dependent proteolysis and the change in the subunit composition of proteasomes in malignant tissue compared with surrounding intact endometrium [1]. In addition, increased activity of proteasome correlates with the stage of disease. These data indicate the involvement of proteasome in the pathogenesis of endometrial cancer, and suggest the ability to register changes in proteasomal system in the early stages of tumor development. Calpaines also are among the most important specific intracellular proteases. It is known that the change in their activity and expression is characteristic of many tumors [10]. Been shown to increase their expression with endometrial cancer compared with hyperplasia, which indicates the possible involvement of these proteinases in the development of malignancy in hyperplastic tissue [8].

The information content of selected criteria, also confirmed the presence of significant differences in activity of proteasomes and kalainov in PE and GE identified in the study, 60 patients (mean age of 57.1±1.7 years) with morphologically verified diagnosis of PE stage I and 50 ill� with hyperplastic processes of the endometrium. All patients with PE was carried out surgical treatment - hysterectomy in a different range. Patients with ET have received organ preservation or organolead surgical treatment.

Risk assessment of PE was based on indicators of the state of proteolytic systems (activity total pool of proteasomes, pools of 26S and 20S, the subunit composition of the proteasome activity kalainov), a clinical-anamnestic data (age, status, menstrual function, the presence of metabolic syndrome, body mass index). The total number of patients included in the analysis, accounted for 40 people. The classification was carried out for groups 1 - low risk of re, 2 - high risk of developing PE. Discriminant analysis included the following stages: initial selection of features to build the model, a set of training samples (the group of patients who are registered attributes that were selected), the selection of informative features and formation on their basis of the group of differential functions, checking the quality of the resulting model. The most important prognostic indicators are presented in table 1 [informative Features of the indicators included in the mathematical model to assess the risk of developing endometrial cancer in patients with hyperplastic processes of the endometrium. Note: Wilks Lambda value is responsible for the implementation of the conditions of pravomernost enable indicator in the model and the significance of the difference values of the discriminant functions in groups what is essential when creating a model; Toler. (tolerance) is a measure for the informativeness of a variable that is a necessary condition for inclusion of a variable in the model; p-level of significance].

Quality assessment of the obtained functions were performed on 20 patients, not included in the main sample. The sensitivity and specificity of the obtained discriminant models were, respectively, 77% and 100%.

Clinical examples

Example 1: Patient A., 60 years old, was admitted to the gynecology Department, research Institute of Oncology of the Russian medical Academy with complaints of pain in the lower abdomen and bloody discharge from the genital tract on the background of 8 years postmenopause. The patient underwent hysteroscopy with endometrial biopsy. Histological conclusion: atypical hyperplasia of the endometrium. Determined the risk of PE in this patient according to the proposed method: was used a mathematical model to predict cancer risk. In the first stage, the calculation was performed discriminant functions Y1 and Y2 in terms of the indicators in this patient.

Y1=0,0285*4,33+0,0091*3,66+0,5429*60-0,0031*6,52-14,6595=18,05

Y2=0,0517*4,33+0,0239*3,66+0,6568*60-0,0072*6,52-21,0753=18,59

In the calculation of the discriminant functions revealed that Y1<Y2. Thus, the patient is classified in a group of "endometrial cancer" with a significance level of p<0.05, which means high risk R�Suite re. The patient performed hysterectomy with appendages, in a morphological study of surgical material on the background of atypical endometrial hyperplasia foci of well-differentiated adenocarcinoma.

Example 2: Patient N., 51 years old, was admitted to the gynecology Department, research Institute of Oncology of the Russian medical Academy with complaints of recurrent episodes of bleeding from the genital tract amid postmenopausal for 3 years. History of endometrial hyperplasia. Morphological conclusion after hysteroscopy with endometrial biopsy: simple typical hyperplasia of the endometrium. Determined the risk of PE in this patient according to the proposed method: used a mathematical model to predict cancer risk. In the first stage, the calculation was performed discriminant functions Y1 and Y2 in terms of the indicators in this patient.

Y1=0,0285*14,33+0,0091*34,7+0,5429*51-0,0031*46,6-14,6595=13,60

Y2=0,0517*14,33+0,0239*34,7+0,6568*51-0,0072*46,6-21,0753=13,66

In the calculation of the discriminant functions revealed that Y1<Y2. Thus, the patient is classified in a group of "endometrial cancer" with a significance level of p<0.05, which means a high risk of developing PE. The patient was performed endometrial ablation. At the control examination after 2.5 years after treatment during the hysteroscopy with endometrial biopsy revealed focal endometrial, intraepithelial�of obayatelnaya, charming hyperplasia (EIN).

Example 3: Patient R., age 51, turned on the Advisory appointment in the Department of gynecology, research Institute of Oncology, Russian Academy complaints once spotting from the genital tract on the background of 2 years postmenopausal. Morphological conclusion after hysteroscopy with endometrial biopsy: simple typical hyperplasia of the endometrium. Determined the risk of PE in this patient according to the proposed method: used a mathematical model to predict cancer risk. In the first stage, the calculation was performed discriminant functions Y1 and Y2 in terms of the indicators in this patient.

Y1=0,0285*22,22+0,0091*126,47+0,5429*51-0,0031*425,4-14,6595=13,49

Y2=0,0517*22,22+0,0239*126,47+0,6568*51-0,0072*425,4-21,0753=13,43

In the calculation of the discriminant functions revealed that Y1 > Y2. Thus, the patient is classified in a group of "endometrial hyperplasia" with a significance level of p<0.05, which indicates a low risk of re. In response to an underlying medical condition and the impossibility of hormone therapy, the patient underwent organ-preserving treatment in the amount of electrosurgical ablation of the endometrium.

Thus, the developed method with the use of mathematical modeling allows to objectify the criteria cancer risk in patients with hyperplastic processes and with a high degree of sensitivity and specific�STI to predict the development of PE, which in turn allows you to plan adequate volume therapy in secondary prevention of PE.

Table 1
IndicatorsWilks' LambdaToler.P-level
The activity of kalainov0,72170,94300,0001
Total proteasome activity0,64770,8735With 0.0013
The age of the patient0,51690,90520,0928
The activity of the 20S pool of proteasomes0,52170,87680,1078
Wilks' Lambda=0.48; p<0,0000

Sources of information

1. Proteasome activity and subunit composition in the tissue of endometrial cancer: relationship with clinical and morphological parameters [text] / L. V. Spirina, I. V. Kondakova, V. D. Koval and others // Bulletin of experimental biology and medicine. - 2012. - No. 4. Pp. 491-494.

2. The multiplicity of forms of the proteasome and not�have a quiet approaches to their separation / B. E. Abramov, V. M. Astakhov, P. A. erokhov etc. / / Izvestiya ran. Biological series. - 2004. - No. 2. Pp. 150-156.

3. RF patent №2310857, IPC G01N 33/68, G01N 33/53. Method of determining indications for hysterectomy in endometrial hyperplasia / Ishchenko A. I. [and others]; applicant and patentee of the State educational institution of higher professional education Moscow medical Academy n. a. I. M. Sechenov. - 2006108570/15; Appl. 20.03.2006; publ. 20.11.2007.

4. Prediction of endometrial cancer in women with endometrial hyperplasia in premenopausal age / Unanyan A. L., I. S. Sidorov, E. A. Kogan, D. V. Baburin // Obstetrics, gynaecology and reproduction. - 2012. - N2. - Pp. 18-24.

5. Prognostic value of determination of sex hormones, their receptors and enzymes in the synthesis and metabolism of estrogens in patients with endometrial cancer / N. In. Bochkarev, I. V. Kondakova, L. A. Kolomiets et al. / / Problems of Oncology. - 2007. - Vol. 53, No. 3. Pp. 315-320.

6. Prognostic criteria cancer risk proliferative processes of the endometrium / A. L. Chernyshev, L. A. Kolomiets, N. G. Kritsky, V. I., Suhodol // Russian journal of cancer. - 2003. - No. 5. - P. 22-25.

7. 26 S proteasome is also been other ideas where production of an authentic major histocompatibility class I-restricted epitope from an intact protein substrate / Ben-Shahar S., Komlosh A., Nadav E. et al. // The J of Biol. Chem. - 1999. - Vol.274. - No.31. - P. 21963-21972.

8. Immunhistochemical analysis for expression of calpaini, calpain 2 and calpastatin in endometrial cancer / D. Salehn, I. Fromberg, C. Haugk, et.al. // Anticancer Res. - 2010. - Vol.30, No. 7. - P. 2837-43.

9. Predicting the risk of endometrial cancer in postmenopausal women presenting with vaginal bleeding: the Norwich DEFAB risk assessment tool / N. Burbos, P. Musonda, I. Giarenis, et al. // Br J Cancer. - 2010. - Vol.102, No.8. - P. 1201-1206.

10. The calpain system and cancer / S. J. Storr, N. O. Carragher, M. C. Frame, et al. // Nature Reviews Cancer. - 2011. - Vol.11. - P. 364-374.

11. The cancer degradome: proteases in cancer biology / D. Edwards, G. Hoyer-Hansen, F. Blasi, B. F. Sloane. - Springer, 2008. - 1000 p.

12. The expression ofsyndecan-1 is related to the risk ofendometrial hyperplasia progressing to endometrial carcinoma / H. Kim, D. S. Choi, S. J. Chang, et al. // J Gynecol Oncol. - 2010. - Vol.21, No. 1. - P. 50-5.

13. Vascular markers CD31, CD34, actin, VEGFB, and VEGFR2, are prognostic markers for malignant development in benign endometrial polyps / B. Hvingel, M. Lieng, B. Roald, A. Ørbo // Open Journal of Obstetrics and Gynecology. - 2012. - Vol.2. - P. 18-26.

A method of assessing the risk of developing endometrial cancer in patients with hyperplastic processes of the endometrium, including hysteroscopy with endometrial biopsy and determination of clinical and anamnestic indicators, characterized in that in the endometrium identify molecular characteristics, such as activity of kalainov, total proteasome activity, the activity of the 20S proteasome, also take into account the age of the patient, and then calculate the discriminant functions Y1 and Y2 by the formula:
Y1=0,0285×X1+0,0091×X2+0,5429×X3-0,0031×X4-14,6595;
Y2=0,0517×X1+0,0239×x2+0,6568×X3-0,0072×X4-21,0753, where
X1 - activity kalainov, ·103U/mg protein,
X2 - total proteasome activity, ·103U/mg protein,
X3 - the age of the patient, l�t,
X4 - 20S activity of a pool of proteasomes, ·103U/mg protein,
and when Y1>Y2 the patient is referred to the group with low risk, and if Y1<Y2 - to the group with a high risk of developing endometrial cancer.



 

Same patents:

FIELD: medicine.

SUBSTANCE: seromucoid concentration is measured in supernatant of a biological fluid aspirated from the nasopharynx of the newborns suffering a generalised form of the intrauterine mono-cytomegalovirus infection or mixed cytomegalovirus infection. If the seromucoid concentration is 0.110-0.140 absorbance units, the early stage of the generalised form of the intrauterine mono-cytomegalovirus infection is diagnosed. If the seromucoid concentration is 0.141-0.171 absorbance units, the early stage of the generalised form of the intrauterine mixed cytomegalovirus infection caused by a combination of the cytomegalovirus and type 1 herpes simplex virus is diagnosed.

EFFECT: using the declared method enables the effective differential diagnosis of the generalised form of the intrauterine mono or mixed cytomegalovirus infection in the newborns.

1 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: predicting toxaemia of pregnancy is ensured by determining 24-hour urine placenta-like alkaline phosphatase and lactoferrin when the woman is 20-22 weeks pregnant, and then 2 weeks later, and the measured placenta-like alkaline phosphatase and lactoferrin of more than 16.0 ng/ml enable estimating potential toxaemia of pregnancy.

EFFECT: method enables predicting potential toxaemia of pregnancy, including the comprehensive assessment of protein risk factors taking into account the clinical presentation and significance of each factor in numeric equivalent.

1 tbl

FIELD: medicine.

SUBSTANCE: invention relates to field of medicine. Claimed is method, including determination of presence or absence of K-rasG12D mutation. Presence of K-rasG12D mutation points to the fact that patient will not respond to treatment with B-Raf inhibitor.

EFFECT: invention provides effective method of identifying patient who does not respond to treatment with B-Raf inhibitor.

6 cl, 34 dwg, 12 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to oncology, and can be used for detecting endogenous intoxication (EI) symptoms in the patients suffering colorectal cancer. That is ensured by studying the complete blood count and deriving an integral intoxication index (III) by formula: EI=(Im+St+Seg+Myel+Plas)*10(Bas+Eos+Lym+Mon)*Er*CC, wherein III is the integral intoxication index; Im is immature leukocytes, %; St is stab neutrophils, %; Seg is segmented neutrophils, %, Myel is myelocytes, %; Plas is plasma cells, %; Bas is basophiles, %; Eos is eosinophils, %; Lym is lymphocytes, %; Mon is monocytes, %; Er is absolute erythrocytes to the power of 1012; CC is an absolute colour characteristics.

EFFECT: invention provides diagnosing the absence (III=4,6) or presence (III=10,4) of the EI symptoms and associated respiratory disorders and enables assessing the therapy efficacy.

FIELD: medicine.

SUBSTANCE: predicting heart rhythm disorder in the patients with pre-excitation syndromes is ensured by measuring patient's blood plasma matrix metalloproteinase-9 (MMP-9), and if the measured value is less than 90.6 ng/ml, the favourable clinical course accompanied by no heart rhythm disorders (HRD) is predicted, whereas the MMP-9 concentration of more than 90.6 ng/ml enables predicting HRD.

EFFECT: method enables predicting HRD in the patients with pre-excitation syndromes by high sensitivity and specificity of the method with its low invasiveness and no counterindications.

3 ex

FIELD: medicine.

SUBSTANCE: technique consists in measuring a pinopod count in the germinal epithelium, an expression level of progesterone and oestrogen alpha receptors in the endometrial stroma, calculating their ratio, and determining an expression level of implantation LIF factor on the 21st-24th day of the menstrual cycle ('implantation window'). If the pinopod count is less than 35%, the expression ratio of progesterone and oestrogen alpha receptors in the endometrial stroma less than 3.5, as well as the LIF expression level of less than 2 points, disturbed fertility associated with uterine myoma is diagnosed.

EFFECT: technique enables applying the differentiated approach to managing the patients planning pregnancy, suffering uterine myoma undeforming the uterine cavity, as well as with myomatous nodule up to 5,0 cm in diameter.

2 ex, 4 dwg

FIELD: medicine.

SUBSTANCE: method involves sampling a biological tissue, introducing a biologically active substance therein, keeping the prepared mixture in a thermostat at 37°C and incubating, performing microscopic examination for identifying the forms of trichomonads. The sampled biological tissue represents the urogenital mucosa that is incubated for 24 h in a nutrient medium. The prepared sediment is separated and added with the biologically active substance nitrosoguanidine in an amount of 500 mcg/ml. That is followed by incubating for 15 min at +37°C; a phosphate buffer with pH 5.6 heated to +37°C is added and centrifuged twice at 1000 rpm for 10 min. The material is placed into the nutrient medium for trichomonad growing, incubated for at least 24 hours, and microscopic examination is used to identify atypical nonflagellate and typical round and amoeboid forms of trichomonads.

EFFECT: using the declared invention enables the effective, fast and reliable identification of typical changed and atypical forms of trichomonads.

3 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: method involves measuring pregnant women's blood anti-cytomegalovirus antibody titre and glutathione reductase activity in erythrocytes of the obstetric patient suffering the cytomegaloviral infection. If the anti-cytomegalovirus antibody titre is 1:1600, whereas the measured glutathione reductase activity is below 4.48±0.22 units/gHb, the aggravated cytomegaloviral infection is diagnosed.

EFFECT: higher diagnostic technique.

FIELD: medicine.

SUBSTANCE: group of inventions relates to medicine, cosmetology, production of food products, vitamins, food supplements, drugs and describes versions of device for realisation of non-invasive potentiometric determination of oxidant/antioxidant activity of biological tissues, which includes device for measuring potentials and double-sided electrode, made in form of plate with similar working surface, covered with electricity-conducting gel, containing mediator system. Electrodes are fixed on biological tissue in such a way that one working surface, playing role of measuring electrode, is in direct contact with biological tissue via gel, second working surface pale role of comparison electrode. Electrodes contact with each other via gel, with oxidant/antioxidant activity being determined by formulae with application of difference between final and initial potentials.

EFFECT: simplification, as well as increase of accuracy and reliability of determination, is achieved.

14 cl, 3 tbl, 4 dwg

FIELD: medicine.

SUBSTANCE: invention represents a method for the prediction of preeclampsia in the second trimester of pregnancy by blood examination, differing by the fact that the activity of acid and neutral proteinases is measured in blood serum of the women 7-8 weeks pregnant; if the activity of acid proteinases is more than 5.6 mcmole/l, while the activity of neutral proteinases is more than 3.9 mcmole/l, the preeclampsia progression in the second trimester of pregnancy is predicted.

EFFECT: higher accuracy and specificity of the method for the prediction of gestational toxicosis.

2 ex

FIELD: medicine, psychiatry.

SUBSTANCE: one should isolate DNA out of lymphocytes of peripheral venous blood, then due to the method of polymerase chain reaction of DNA synthesis one should amplify the fragments of hSERT locus of serotonin carrier gene and at detecting genotype 12/10 one should predict the risk for the development of hallucino-delirious forms of psychoses of cerebro-atherosclerotic genesis.

EFFECT: more objective prediction of disease development.

3 ex

FIELD: medicine, urology.

SUBSTANCE: one should conduct subcutaneous prevocational tuberculin test and, additionally, both before the test and 48 h later it is necessary to perform the mapping of prostatic vessels and at decreased values of hemodynamics one should diagnose tuberculosis. The information obtained should be documented due to printing dopplerograms.

EFFECT: more reliable and objective information.

1 ex, 1 tbl

FIELD: molecular biology.

SUBSTANCE: the suggested innovation deals with the fact that nucleic acids should be isolated directly out of the sample without pipetting stage but with the help of interconnected reservoirs being prepared beforehand. The above-mentioned vessels should be applied either separately or being interconnected according to standard microtitrating format. The sample should be mixed with a lyzing buffer and nucleic acids are bound with matrix in closed system including, at least, two interconnected reservoirs. Forced movement of sample's mixture and buffer back and forth from one reservoir into another one for several times through narrow passage provides their thorough intermixing. The method provides quick and safe isolation of nucleic acids.

EFFECT: higher efficiency.

44 cl, 4 dwg, 1 ex

FIELD: medicine, phthisiology, microbiology.

SUBSTANCE: diagnostic material is poured preliminary with chlorohexidine bigluconium solution, homogenized, kept at room temperature for 10-12 h and centrifuged. Precipitate is poured with Shkolnikova's liquid medium, incubated at 37oC for 3 days, supernatant part of Shkolnokova's medium is removed, fresh Shkolnikova's medium is added, and precipitate is stirred and inoculated on the dense cellular egg media. Sensitivity of the strain is determined in 3 weeks by the presence of growth in the control tube only. Invention provides enhancing precision and reducing time for assay. Invention can be used in assay for medicinal sensitivity of tuberculosis mycobacterium.

EFFECT: improved assay method.

3 ex

FIELD: medicine, biotechnology, pharmacy.

SUBSTANCE: invention relates to agents used for treatment of pathological states associated with disorder of synthesis of neuromediating substances. Method involves the development of pharmaceutical composition and a method for it preparing. Pharmaceutical composition represents subcellular synaptosomal fractions: synaptic membranes, "light" synaptosomes and "heavy" synaptosomes prepared from gray matter of cerebral hemispheres from experimental animals based on the goal-seeking modification of humoral mediators of nerve endings transformed to synaptosomes in development and regression of malignant processes. The composition provides inhibiting the growth of tumor cells, to elevate span-life of patients with ascite Ehrlich's sarcoma, breast adenocarcinoma Ca-755, Wolker's carcinosarcoma-256.

EFFECT: valuable medicinal and anti-tumor properties of composition.

12 cl, 3 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: method involves carrying out microscopic examination of blood serum samples taken from femoral vein and cubital vein. Femoral vein sample is taken on injured side. The examination is carried out before and after treatment. The blood serum samples are placed on fat-free glass slide in the amount of 0.01-0.02 ml as drops, dried at 18-30°C for 18-24 h. The set of pathological symptoms becoming larger or not changed after the treatment in comparison to sample taken before treatment, and morphological picture of samples under comparison taken from the cubital vein showing no changes or being changed to worse, the treatment is considered to be effective.

EFFECT: enabled medicamentous treatment evaluation in course of treatment to allow the treatment mode to be changed in due time; avoided surgical intervention (amputation); retained active life-style of aged patients.

4 dwg

FIELD: medicine, clinical toxicology.

SUBSTANCE: at patient's hospitalization one should gather the data of clinical and laboratory values: on the type of chemical substance, patient's age, data of clinical survey and laboratory values: body temperature, the presence or absence of dysphonia, oliguria being below 30 ml/h, hemoglobinuria, erythrocytic hemolysis, exotoxic shock, glucose level in blood, fibrinogen and creatinine concentration in blood serum, general bilirubin, prothrombin index (PTI), Ph-plasma, the state of blood clotting system. The state of every sign should be evaluated in points to be then summed up and at exceeding the sum of points being above "+20" one should predict unfavorable result. At the sum of "-13" prediction should be stated upon as favorable and at "-13" up to "+20" - prediction is considered to be doubtful.

EFFECT: higher accuracy of prediction.

2 ex, 3 tbl

FIELD: medicine, juvenile clinical nephrology.

SUBSTANCE: disease duration in case of obstructive pyelonephritis should be detected by two ways: either by detecting the value of NADPH-diaphorase activity, as the marker of nitroxide synthase activity in different renal department and comparing it to established norm, or by detecting clinico-laboratory values, such as: hemoglobin, leukocytes, eosinophils, urea, beta-lipoproteides, lymphocytes, neutrophils, the level of glomerular filtration, that of canalicular reabsorption, urinary specific weight, daily excretion of oxalates, arterial pressure, and estimating their deviation against average statistical values by taking into account a child's age.

EFFECT: higher efficiency of detection.

7 dwg, 1 ex, 6 tbl

FIELD: medicine, urology.

SUBSTANCE: the present innovation deals with differential diagnostics of prostatic cancer and other prostatic diseases at the stage of primary inspection. The method includes the detection of PCA and calculation of probability coefficient for prostatic cancer (PCC) by the following formula: where e - the foundation of natural logarithm (e=2.718…), PCA - the level of total blood PCA in ng/ml, V - patient's age in years. At PCC value being above 0.2 one should diagnose prostatic cancer and to establish final diagnosis one should perform polyfocal prostatic biopsy. The method enables to increase accuracy of diagnostics at decreased number of unjustified prostatic biopsies.

EFFECT: higher efficiency of diagnostics.

2 ex

FIELD: medicine, biology.

SUBSTANCE: invention relates to nutrient medium used for accumulation of cells for the following cytological and/or immunocytochemical analysis carrying out. Invention relates to medium containing salts NaCl, KCl, anhydrous CaCl2, MgSO4 x 6 H2O, MgCl2 x 6 H2O, Na2HPO4 x 2 H2O, KHPO4, NaHCO3, and also glucose and Henx's solution, 10% albumin solution and polyglucin taken in the ratio 1:1:1. Invention provides enhancing the preservation of cells.

EFFECT: improved an valuable properties of nutrient medium.

3 ex

Up!