Method of monitoring fibrous process in liver of patients with chronic hepatitis c

FIELD: medicine.

SUBSTANCE: invention relates to field of medicine, in particular hepatology and infectious diseases, and can be used for determination of stage of fibrous process in monitoring of patients with chronic hepatitis C. To realise method levels of blood serum cytokins are determined in patients with chronic hepatitis C with diagnosed by means of biopsy or other non-invasive method stage of fibrous process 2 times a year, with further calculation of cytokine profile integral index (CPII) on their basis, at initial stage F0 growth of CPII higher than -8 testifies to debut of fibrous changes in liver (transition to stage F1), at initial stage F1, drop of CPII below -10 testifies to transition to stage F2, at initial stage F2 growth of CPII higher than -3 testifies to transition of fibrosis to stage F3, at initial stage F3 drop of CPII below -3 testifies to development of cirrhosis.

EFFECT: determination of stage of fibrous process in monitoring of patients with chronic hepatitis C.

3 ex, 1 tbl, 5 dwg

 

The invention relates to medicine, in particular Hepatology and infectious diseases, and can be used to determine the stage of the fibrotic process in the monitoring of patients with chronic hepatitis C. For carrying out the method in patients with chronic hepatitis C is installed in the biopsy or other non-invasive stage fibrotic process 2 times a year to determine the integral indicator of cytokine profile (IPSP), then at the initial stage F0 growth IPCP above -8 indicates the onset of fibrotic changes in the liver (transition to stage F1), at an initial stage F1 drop IPCP below -10 indicates the transition to the stage of F2, at the initial stage F2 growth IPCP above -3 indicates the transition of fibrosis stage F3 at the initial stage F3 drop IPCP below -3 shows the development of liver cirrhosis METAVIR.

Infection with hepatitis C virus (HCV) is characterized by a very high incidence throughout the world (about 200 million people), a high level of chronicity of the process with unfavorable outcomes in cirrhosis or liver cancer [1, 2, 3]. From a socio-economic perspective of chronic hepatitis C requires a significant investment on dispensary observation of patients when accompanied by expensive diagnostic monitoring [3, 4]. With clinical positions "gold standard" of diagnosis�IKI fibrotic process in the liver biopsy unsafe for the patient [5, 6], and numerous ways for non-invasive diagnosis of liver fibrosis is not always accurate, require a combination of several tests [7, 8, 9, 10] and in need of new effective diagnostic techniques that best reflects the pathogenesis of fibrotic changes in the liver and are readily available for monitoring patients with both methodological and economic points of view.

It is known that fibrotic processes in the liver is very closely associated with immunological changes in patients with chronic hepatitis C and largely depend on the ratio in the patient cytokines profibrotic and protivovirusnogo action. In particular, it was shown that the progression of liver fibrosis has been fostered cytokines associated with CD4+ Tx2immune response (IL-4, IL-5, IL-13, IL-21), while protivovirusnymi properties have Tx1-associated cytokines IFN-γ and IL-12 [11]. Particularly pronounced provironum action is characterized by transforming growth factor β (β), through which the receptors on the membrane of myofibroblasts greatly affects the transcription of genes responsible for the synthesis of procollagen I and III [11, 12]. In turn, the secretion β macrophages effectively induced IL-13 [13]. A very important function of the blockade of fibrotic changes in the liver performs IL-10, which show�about numerous models, including with HCV [14].

In this regard, the aim of the present invention was the development of a new method of monitoring fibrotic process in the liver in patients with chronic hepatitis C based on the integral evaluation of the cytokine profile of blood serum in this disease.

Currently, there are ways to assess fibrotic process on the basis of cytokine status in patients with chronic hepatitis C. Thus, the known methods for predicting cirrhosis of the liver on the background of chronic hepatitis C from the use of genetic methods of research, in particular, by analyzing the DNA of patients for the presence of gene polymorphisms of cytokines [15, 16]. Analysis of the level of cytokines in the diagnosis of liver fibrosis may be part of a complex immunological tests designed for non-invasive diagnosis of liver fibrosis in patients with chronic hepatitis C [17].

Both methods for its availability significantly inferior to the method of monitoring the severity and prognosis of cirrhotic changes in the liver in chronic hepatitis C based on the definition in the serum levels of various cytokines [18], which can serve as an analogue of the claimed invention. For implementing the method in the serum define the content of cytokines IL-4, IL-10, IL-R, and TNF-a. When IL-4 above 5.6 PG/ml, IL-10 above 35 PG/ml, and TNF-above 14 ng/ml and IL-R above 8.5 PG/ml set cash�e pronounced liver fibrosis. Common analogue of the claimed invention is in methods of immunological test based on determining the level of serum cytokines by the method of enzyme-linked immunosorbent assay, and also in carrying out research under the control of liver biopsy.

The advantage of both methods can be considered their economic feasibility and availability, as the definition of the levels of cytokines falls into the category of routine methods of immunoassay based on enzyme-linked immunosorbent assay and in principle can be carried out on the basis of domestic test systems, which significantly reduces their cost.

Along with this, the analogue has certain disadvantages, one of which is the ability of a single definition of marked liver fibrosis in a patient with no prospect of its use for long-term monitoring of the patient's condition CHC and hence the identification of the indications to perform antiviral therapy based on the severity and extent of progression of the fibrotic process. In addition, using the method establishes a pronounced fibrosis without the possibility of clear identification of each stage of the fibrotic process. The disadvantage of this method is associated with a methodical approach to its solution because it is based on setting the level of the individual qi�of okinow in serum without consideration of the contribution ("weight") of each cytokine in the development of individual stages of fibrotic changes in the liver.

In this regard, a feature of this invention is that to achieve the technical result of using a different set of tested cytokines in serum by calculation on the basis of a particular factor is an integral indicator of cytokine profile (IPSP), which takes into account the contribution of each cytokine in the development of the fibrotic process in the liver at each stage of its development in patients with chronic hepatitis C.

The object of the research included 66 people with verified diagnosis of chronic hepatitis C, which was previously performed liver biopsy and histologically established stage of liver fibrosis according to the METAVIR score [19]. As a result, all the patients studied in accordance with the stages of fibrosis of the liver was divided into 5 subgroups: stage F0 (no fibrosis) to 4 guests, on stage F1 (early fibrotic changes) - 25 people on stage F2 (pronounced fibrosis) - 7 people on stage F3 (significant fibrosis) - 9 people on stage F4 (cirrhosis) 21. Control was a group of 17 healthy people (donors).

Each of the surveyed determined the level of this cytokine serum: interferon α (IFN-α), interferon γ (IFN-γ), interleukin-10 (IL-10), interleukin-12 (IL-12), interleukin-13 (IL-13), tumor necrosis factor α (TNF), transforming growth factor � (β).

Material for the study was venous blood of patients in an amount of 5 ml, which were taken by syringe from the cubital vein, centrifuged at 3000 rpm in the cold for 10 minutes, the serum was poured 0.5 ml ependorf, were frozen and stored until use at -70°C. enzyme-linked Immunosorbent assay (ELISA) was performed from serum samples using a tablet photometer "OPSYS MR" (reader) of the company "THERMOLABSYSTEMS" (Finland) in accordance with instructions for use of the equipment and a set of monoclonal antibodies. The latter included: anti-IFNα MKAT (Vector-best, Russia) to determine the level of IFN-α; anti-IFNγ MKAT (Vector-best, Russia) to determine the level of IFN-γ; anti-IL-10 MKAT (Vector-best, Russia) to determine the level of IL-10; anti-IL-12 MKAT (Vector-best, Russia) to determine the level of IL-12; anti-IL-13 MKAT (Vector-best, Russia) to determine the level of IL-13; anti-TNFα MKAT (Vector-best, Russia) to determine the level of TNF; anti-TGFβ MKAT (Biosource, Canada) to determine the level β.

Statistical data processing was performed on the basis of the statistical software package SPSS 17.0. A comparative analysis was carried out using non-parametric statistics using the Mann-Whitney test in accordance with the distribution data. Statistical definition the formula for calculating IPCP was carried out by plotting a linear regression equation. Check �Heathrow on the received data according to the developed criteria of diagnosis was carried out based on the calculation of 95% confidence intervals in accordance with the stages of liver fibrosis METAVIR, certain biopsy. The ratio of sensitivity and specificity of the developed tests of separate stages of hepatic fibrosis were established by linear regression with plotting the ROC curve and calculating the area under the curve - AUROC[20, 21, 22].

The main differences between cytokine profiles in patients with chronic hepatitis C at different stages of the fibrotic process was detected mainly in comparison with the group of healthy individuals (table 1). In this case, each stage was characterized by its reliable shifts, which involved almost all tested cytokines. However, the differences between stages were isolated and did not allow to identify individual stage.

All obtained data were used for the regression analysis with the construction of the linear regression equation. The result was obtained integral coefficient cytokine profile (IPSP), which was calculated by the formula 1.

Formula 1

IPCP=5,052-0,192*[IFN]-0,006*[IFN-γ]+0,169*[IL-10]+0,044*[IL-12]-0,146*[IL-13]-0,145*[TNF]-0,050*[β],

where [IFN] - the level of IFN-α in PG/l in the serum of the patient, [IFN-γ] - the level of IFN-γ in PG/l in the serum of the patient, [IL-10] - the level of IL-10 in PG/l in the serum of the patient, [IL-12] - level IL-12 in PG/l in the serum of the patient, [IL-13] - level IL-13 PG/l in the serum of the patient, [TNF - the level of TNF-a in PG/ml in the serum of the patient, [β] - level β in PG/l in the serum of the patient.

The relevant calculations IPCP for each patient, expressed as ranges of values with 95% confidence intervals of this ratio, shown in Fig.1.

As can be seen from the figure, if the study was carried out only once during the initial examination of the patient, to determine the stage of liver fibrosis would be rather difficult, since data on the different stages can be "layered" on each other. However, when monitoring of the patient and the knowledge of the initial stage of liver fibrosis using IPCP to assess the stage is diagnostically highly effective.

So, in the case of the initial stage of the F0 rise IPCP above -8 indicates the onset of fibrotic changes in the liver, that is, the fibrous transition process to the stage F1. The diagnostic value of this test, evaluated by constructing ROC curve and compute the area under the curve (AUROC), is quite high because AUROC=0,765, as illustrated in Fig.2.

At the initial stage F1 drop IPCP below -10 indicates the transition process to the stage of pronounced liver fibrosis F2, and the diagnostic value of the test, judging by the ROC curve characterizing this transition is largest IPCP is even more variable AUROC=0,824 (Fig. 3).

When the original study� F2 growth IPCP above -3 indicates the transition of fibrosis stage significant fibrosis F3 as can be seen in Fig. 4. This transition, estimated largest IPCP is diagnostically significant, while the diagnostic accuracy of the test is approaching the absolute, since AUROC=1 (transition from one value range to another is observed in 100% of cases).

At the initial stage F3, on the contrary, the fall IPCP below -3 shows the development of liver cirrhosis - F4 (Fig. 5) with relatively high AUROC=0,824.

The method is illustrated by the following figures.

Fig.1. 95% confidence intervals (95% CI) values IPCP in patients with chronic hepatitis C at different stages of liver fibrosis.

Fig.2. ROC-curve IPCP to go CHC patients with stage F0 to F1 stage.

Fig.3. ROC-curve IPCP to go CHC patients with stage F1 stage F2.

Fig.4. ROC-curve IPCP to go CHC patients with stage F2 to the stage of F3.

Fig.5. ROC-curve IPCP to go CHC patients with stage F3 on stage F4.

The method is illustrated by the following clinical examples.

Example 1.

Patient A., 37 years old, is observed as outpatients in the EMC Hepatology center for Infectious clinical hospital №1 with a diagnosis of Chronic hepatitis With severe activity, minimal fibrosis (stage 1 according to the METAVIR score)".

According to history the first antibody to hepatitis C virus identified in April 2011 in the clinical examination in polyclinic №49. The result was confirmed by a study � may 2011 for anti-HCV and detection of HCV RNA. The patient addressed in the SDS No. 1, diagnosed with genotype 1b HCV. For biochemical analysis of blood revealed the activity level of ALT 47 U/L. Treatment with interferon, pegylated interferon, ribavirin during the life of the patient had not received.

Related other chronic diseases denies. According to the patient's allergies to medications no. From illness by history: sepsis at the age of 2 years, (a transfusion of blood plasma), tonsilectomy in 1983, rheumatism at the age of 10. Working as a lawyer.

The patient was recommended a biopsy of the liver 16.06.2011 G. biopsy diagnosed chronic hepatitis With severe activity, minimal fibrosis (stage 1 according to the METAVIR score).

The results of the study of cytokine status for 3 days before performing a liver biopsy (13.06.2011): IFN - 4,75 PG/l, IFN-γ - 40 ng/l, IL-10 - 11,67 ng/l, IL-12 - the 8.25 ng/l, IL-13 is 74.5 PG/l, and TNF - 0, β - 8,72 PG/L. IPCP=-7,24, which corresponds to stage F1 and was confirmed by the results of subsequent liver biopsy.

Since January 2012, the patient was noted level rise Alat: 31.01.2012. - 163 U/l and was recommended to undergo antiviral therapy. Before starting therapy in January 2012, the patient underwent a comprehensive examination. During the physical examination - without features According to abdominal ultrasound echographic pattern corresponds to the diffuse changes of the liver by the type of chronic hepatitis without exacerbation, CWR increased to 162 mm.

The results of the study of cytokine status: IFN - 8,93 PG/l, IFN-γ - 110 ng/l, IL-10 - 14,11 ng/l, IL-12 - at 11.57 ng/l, IL-13 - 83,8 PG/l, and TNF-a was 5.04 PG/ml, β - 11,0 PG/L. IPCP=-13,89 that evidence in favor of the transition of liver fibrosis stage F2. The patient was assigned a procedure transient elastography, which was installed the average density of the liver was 8.8 kPa, which confirmed the patient had stage F2, testified to the progression of the fibrotic process in the last six months and confirmed the diagnostic value of monitoring of the patient by examining its cytokine profile.

Example 2.

Patient P., age 31, is observed as outpatients in the EMC Hepatology center for Infectious clinical hospital №1 with a diagnosis of Chronic hepatitis With moderate activity." According to the case history and discharge from hospital for the first time, antibodies to hepatitis C virus identified in August 2010 in the survey in the gynecological Department of the hospital №70. In September 2010, identified HCV RNA, genotype 1b. Further analysis in September 2011, according to the results of fibroelastoma liver diagnosed second stage of liver fibrosis (7,8 kPa). Treatment with interferon, pegylated interferon, ribavirin during the life of the patient did not receive, do not take any drugs �permanent. For the last 6 months. there is the rise of Alat level: 16.08.2011 G. - 46 U/L, 05.01.2012. - 49 U/L. Related, and other chronic diseases denies. According to the patient allergies to medications no. During the life of the ill SARS and influenza, inpatient treatment for incomplete spontaneous abortion in pregnancy small term. Is a Manager.

Before you start antiviral therapy in August 2011, the patient underwent a comprehensive examination. During the physical examination were unremarkable. According to abdominal ultrasound: echoprint diffuse changes of the liver by the type of hepatosis-hepatitis.

The results of the study of cytokine status (11.08.2011): IFN - 2,66 PG/l, IFN-γ - 214 ng/l, IL-10 - 7,70 ng/l, IL-12 - 16,34 ng/l, IL-13 to 8.1 PG/l, TNF - a 6.8 PG/ml, β was 7.2 PG/L. IPCP=-8,81 that corresponds to phase F2.

After a few months the patient began to receive therapy with pegylated α2b 1 time per week subcutaneously and ribavirin at a dose calculated according to body weight. Viral load before treatment was of 1.09×106 IU/ml HCV RNA, after a month of therapy was determined by the viral load of 7.14×102 IU/ml HCV RNA. Thus, rapid virologic response has not been achieved.

In February 2012, the patient was performed a liver biopsy, was installed stage F2 by METAVIR scale and was performed in parallel re ISS�adowanie cytokine status.

The results of the study of cytokine status (on 03.01.2012): IFN - 10,23 PG/l, IFN-γ - 30 ng/l, IL-10 - 10,56 ng/l, IL-12 - 8,40 ng/l, IL-13 was 78.5 PG/l, and TNF - 0, β - the 11.04 PG/L. IPCP=-8,57 that corresponds to phase F2. As can be seen, the levels of individual cytokines, being changeable parameters, in this patient after 6 months is quite changed, but the value IPCP, despite this, remained stable and still testified to the presence in a patient with stage F2.

Example 3.

Patient K., 43 years old, is observed as outpatients in the EMC Hepatology center for Infectious clinical hospital №1 with a diagnosis of Chronic hepatitis With severe activity, pronounced fibrosis (stage 3 according to the METAVIR score)". According to history the first antibody to hepatitis C virus identified in November 2010, when examined in hospital №57, where the patient was treated about toxico-allergic dermatitis. For biochemical analysis of blood revealed a high level of ALT activity is more than 600 µmol/L. min. In January 2011, identified HCV RNA, genotype 1b.

These determine the stage of liver fibrosis were contradictory. Further analysis according to transient elastography revealed fibrosis stage 4 (16,3 kPa). According to the results of a liver biopsy diagnosed chronic hepatitis With significant fibrosis (stage 3 according to the METAVIR score).

To resolve the disputed issue was the dash�eno determination of cytokine status and obtained the following result: IFN - 12,5 PG/l, IFN-γ - 55,0 ng/l, IL-10 and 18.2 ng/l, IL-12 - 6,0 PG/l, IL-13 to 21.3 ng/l, TNF - a 7.3 PG/ml, β to 5.0 PG/L. IPCP=-1,73, which corresponds to stage F3 and biopsy confirms the data, but not transient elastography.

The results of the determination of levels of cytokines in different stages of liver fibrosis compared with the control are presented in table 1.

td align="center"> 0,591
Table 1
Average values of the levels of cytokines in blood of patients with chronic hepatitis C at different stages of liver fibrosis and in healthy persons
Stage of liver fibrosisTested cytokinesPatients with CHCHealthy people, n2=17p1p2
IFN-α, PG/l5,60±4,637,70±3,87Of 0.531-
IFN-γ PG/l45,0±90,085,6±56,00,136-
F IL-10, PG/l8,16±1,6212,3±3,870,066-
n1=4IL-12, PG/l10,6±1,5912,7±6,690,964-
TNF-a, PG/ml5,34±4,052,45±3,360,209-
IL-13, PG/l98,8±29,664,6±40,20,044-
β, PG/l5,90±0,847,06±2,080,234-
IFN-α, PG/l8,08±4,037,70±3,870,5470,363
IFN-γ PG/l 27,4±35,785,6±56,00,0010,870
F1IL-10, PG/l10,9±5.08 mm12,3±3,870,1780,442
n1=25IL-12, PG/l10,1±5,1012,7±6,690,0950,617
TNF-a, PG/ml3,33±5,392,45±3,360,9840,735
IL-13, PG/l57,0±47,164,6±40,20,8900,158
β, PG/l10,6±2,257,06±2,08<0,000,520
IFN-α, PG/l8,62±4,277,70±3,870,427
IFN-γ PG/l92,0±67,785,6±56,00,9410,221
F2IL-10, PG/l12,0±3,4112,3±3,870,9240,958
n1=7IL-12, PG/l12,6±3,4312,7±6,69Of 0.5310,634
TNF-a, PG/ml2,96±3,492,45±3,360,9210,777
IL-13, PG/l65,0±38,264,6±40,20,5300,751
β, PG/l10,5±1,467,06±2,080,0020,610
IFN-α, PG/lA 10.4±2,547,70±3,870,0350,999
F3IFN-γ PG/l52,4±4,4585,6±56,00,0820,564
n1=9IL-10, PG/l18,8±14,412,3±3,870,1780,564
IL-12, PG/l8,93±2,8012,7±6,690,0490,564
TNF-a, PG/ml7,17±3,442,45±3,360,0110,999
IL-13, PG/l41,1±44,364,6±40,20,1450,999
�β, PG/l5,77±2,567,06±2,080,1240,043
IFN-α, PG/l10,1±4,407,70±3,870,2100,621
IFN-γ PG/l10,6±24,885,6±56,00,0010,348
F4IL-10, PG/l11,8±3,3112,3±3,870,6070,399
n1=21IL-12, PG/l13,0±7,7612,7±6,690,9990,999
TNF-a, PG/mlOr 3.28±4,562,45±3,360,7140,939
IL-13, PG/l55,6±36,0 64,6±40,20,9340,573
β, PG/l10,7±3,427,06±2,080,0010,505
Note: p1- probability of differences between parameters in patients and healthy individuals, p2- probability of differences between parameters in human patients at different stages of fibrosis with the previous stage, n1- the number of patients, n2- the number of healthy, grey color marks the significance of differences according to the Mann-Whitney test at p<0,05

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Method of monitoring fibrotic process in the liver in patients with chronic hepatitis C, including a study of the content in the serum interferon α and γ, interleukin-10, -12, -13 and tumor necrosis factor α, transforming growth factor β, characterized in that the calculated Integral Indicator of Cytokine Profile (IPSP) by the formula: IPCP=5,052-0,192*[IFN]-0,006*[IFN-γ]+0,169*[IL-10]+0,044*[IL-12]-0,146*[IL-13]-0,145*[TNF]-0,050*[TFR], where [IFN] - the level of IFN-α in PG/l in the serum of the patient, [IFN-γ] - the level of IFN-γ in PG/l in the serum of the patient, [IL-10] - the level of IL-10 in PG/l in the serum of the patient, [IL-12] - level IL-12 in PG/l in the serum of the patient, [IL-13] - level IL-13 PG/l in the serum of the patient, [TNF-a] - level �α in PG/ml in the serum of the patient, [β] - level β in PG/l in the serum of the patient; and, at the initial stage F0 growth IPCP>-8 assess how the transition to stage F1, at an initial stage F1 drop IPCP<-10 - as the shift to F2 at the initial stage F2 growth IGMP>-3 is how the transition of fibrosis stage F3 at the initial stage F3 drop IPCP<-3 - as the development of cirrhosis of the liver, fibrosis of the liver evaluated by the METAVIR score.



 

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2 dwg, 2 ex

FIELD: medicine.

SUBSTANCE: invention aims at treating drug-induced dry eye syndrome (DI-DES). Treating DI-DES implies taking the past medical history, measuring tear production and eye xerosis values reduced and increased respectively in relation to the norm. Unpreserved ocular hypotensive medications are prescribed in the patient. Unpreserved artificial tears are also applied. The lachrymal fluid is analysed by a multicytokine technique. If the analysis shows increased concentrations of proinflammatory cytokines - interleukin-6, interleukin-8, interleukin-12, Th-1 - interleukin-2, interferon-gamma, and Th-2 - interleukin-4, by min 30% in relation to the patient's age norm, a chronic immune ocular inflammation is detected. That requires transpalpebral Blepharogel-1 phonophoresis and 1% hydrocortisone ointment phonophoresis on the sub-mastoidal region from both sides; the therapeutic course is 8-10 daily procedures.

EFFECT: optimal conditions for diagnosing and reasoned differentiated therapy of DI-DES that enables prescribing the pathogenetically reasoned therapy in due time and increasing the efficacy of the therapeutic exposure.

1 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: in a premature baby the concentration of neuron-specific enolase (NSE), concentration of a brain-derived neurotrophic factor (BDNF), concentration of a vascular-endothelial growth factor (VEGF) in umbilical blood and concentration of the vascular-endothelial growth factor (VEGF) in peripheral blood are determined on the basis of the enzyme immunoassay of umbilical and peripheral blood serum on the 7-th day of life, a prognostic index (PI) is calculated by formula: PI=-0.007×X1+0.006×X2-0.05×X3+0.0004×X4-3.9, where X1 is VEGF content in umbilical blood at birth (ng/ml); X2 is VEGF content in peripheral blood on the 7-th day of life (ng/ml); X3 is NSE content in umbilical blood (mcg/l); X4 is BDNF content in umbilical blood (ng/ml); Const=-3.9. When PI is higher than 0, a conclusion about the absence of risk of occlusive posthaemorrhagic hydrocephalus formation is made, and if PI is lower than 0, a high risk of the said pathology development is predicted.

EFFECT: invention makes it possible to increase the efficiency of prediction of occlusive posthaemorrhagic hydrocephalus formation in the premature children with an extremely low body weight at birth.

2 ex

FIELD: medicine.

SUBSTANCE: group of inventions relate to medicine and deals with method of diagnosing neurodegenerative disease in individual, including the following stages (i) determination of one or several parameters, selected from group, consisting of 3ab40 or value of calculated parameter, selected from group, consisting of 2ab40+3ab40, 2ab40+3ab40+2ab42+3ab42 and 1ab40+2ab40+1ab42+2ab42; (ii) comparison of parameter value with standard value, corresponding to value of said parameter in standard sample; and (iii) diagnostics of neurodegenerative disease, in case if increase of parameter value in comparison with standard value is observed. Group of inventions also deals with method of detecting stage, preceding neurodegenerative disease, method of differentiating neurodegenerative disease from stage, preceding said neurodegenerative disease.

EFFECT: group of inventions provide high sensitivity and specificity of detection methods.

13 cl, 12 ex, 14 dwg, 12 tbl

FIELD: medicine.

SUBSTANCE: patient's synovial fluid is sampled, and patient's supernatant chemokines CXCL9/MIG, CXCL10/IP-10 and CXCL11/ITAC are measured. A diagnosis of rheumatoid arthritis is diagnosed, if at least one of chemokines exceeds a threshold; the chemokine thresholds make 2625.8 pg/ml for CXCL9/MIG, 3108.2 pg/ml for CXCL9/MIG and 32.4 pg/ml for CXCL11/ITAC respectively. If the measured values are below the thresholds for three chemokines at the same time, the absence of rheumatoid arthritis and potential osteoarthrosis are stated.

EFFECT: using the given method enables differentiating rheumatoid arthritis and osteoarthrosis by measuring specific markers in the synovial fluid taken from the location directly.

2 ex

FIELD: medicine.

SUBSTANCE: invention relates to method of diagnosing rheumatoid arthritis, method of determining therapeutic agent for treatment of rheumatoid arthritis and set for realisation of methods. Methods are characterised by the fact that include stage of measuring amount of talin in plasma or serum of animal subject. Said measurement is carried out, for instance, by immunologic method with application of antibody, binding with talin. If amount of talin is higher than its average value in control subject without rheumatoid arthritis, rheumatoid arthritis is diagnosed in subject. In case of reduction of talin amount after introduction of therapeutic agent in comparison with amount of talin before introduction, therapeutic effect is stated. Set in accordance with claimed invention contains solid-phase carrier, to which antibody, binding with talin, is attached.

EFFECT: increased efficiency of diagnostics.

9 cl, 4 tbl, 4 ex, 3 dwg

FIELD: medicine, ophthalmology.

SUBSTANCE: in lacrimal liquid one should detect the content of interleukin 8 (IL-8) and that of interleukin 1 beta (IL-1β) to calculate prognostic coefficient (PC) due to dividing the first value by the second one by the following formula: At PC value being below 10.0 one should predict favorable disease flow, and at PC value being above 10.0 - unfavorable flow.

EFFECT: higher accuracy of prediction.

2 ex

FIELD: medicine, medicinal microbiology.

SUBSTANCE: method involves growing microorganism culture to be studied in solid nutrient medium followed by preparing microbial suspension and its incubation in the presence of lactoferrin. Control sample is prepared in parallel series. Control and experimental samples are incubated, supernatant is removed from bacterial cells and lactoferrin concentration is determined in supernatant of experimental and control sample by immunoenzyme analysis. Then anti-lactoferrin activity is calculated by difference of concentrations of residual lactoferrin in experimental and control samples. This method provides enhancing the sensitivity and precision in carrying out the quantitative evaluation of anti-lactoferrin activity in broad spectrum of microorganisms that is urgent in diagnosis and prognosis of diseases with bacterial etiology. Invention can be used in determination of persistent indices of microorganisms for assay of their etiological significance in pathological processes.

EFFECT: improved assay method.

3 tbl, 3 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

FIELD: medicine, cardiology.

SUBSTANCE: in peripheral blood one should detect the level of CD95(+) and CD16(+) neutrophilic granulocytes and at combination of increased level of CD95(+) neutrophilic granulocytes by 4 times and more and CD16(+) neutrophilic granulocytes by 0.6 times against the norm with ECG signs of myocardial infarction one should predict lethal result of large-focal myocardial infarction.

EFFECT: higher accuracy of prediction.

FIELD: medicine, parasitology.

SUBSTANCE: one should carry out immunoenzymatic assay to detect diagnostic optic density and that of labeled immune complex in a plot's hole with tested serum measured in conventional units at wave length being 492 nm. One should calculate coefficient of antibodies concentration measured in conventional units by the following formula: CAC = (Odtsh - Odd) x 100, where CAC - coefficient of antibodies concentration, Odtsh - optic density of the hole with tested serum, Odd - diagnostic value of optic density, 100 - coefficient of serumal dilution. By CAC value one should detect the titer of antibodies to Lamblia intestinalis antigens to interpret results of the trial. The method enables to study the dynamics of disease flow.

EFFECT: higher efficiency and accuracy of diagnostics.

1 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: the present innovation deals with studying and treating diseases of inflammatory, autoimmune and degenerative genesis. One should perform sampling of heparinized blood followed by its sedimentation to obtain blood plasma with leukocytes and centrifuging to isolate the latter which are washed against erythrocytic and serumal admixtures, and, also, it deals with calculating the number of cells in samples out of leukocytic suspension after incubation (B) for 1.5 h at 37 C in holes of plastic microplotting board, out of leukocytic suspension one should additionally prepare two samples, one should be applied to calculate total number of leukocytes before incubation (A), the second sample undergoes incubation at the same mode at addition of autoserum to calculate the number of cells remained after incubation (C). One should state upon adhesive properties of leukocytes by the index of spontaneous adhesion (D), where D=(A-B)/B.100%, and effect for enhanced cellular adhesion under the impact of autoserum should be detected by the value of K=(B-C)/C.100% at K ≥ 30%, where B - C - the number of cells undergone additional adhesion after addition of autoserum. The present innovation widens functional possibilities of the suggested method due to obtaining additional values depicting adhesive properties of blood leukocytes.

EFFECT: higher accuracy of detection.

FIELD: medicine, immunology.

SUBSTANCE: one should carry out reaction of blast-transformation, detect proliferation of T-lymphocytes activated with antibodies to CD3 in the presence of interleukin-7 (ACT IL-7) and in the presence of interleukin-7 and dexametazone (ACT IL-7 D), calculate the index for dexametazone action as the ratio of ACT IL-7 to ACT IL-7 D, moreover, the value of dexametazone action index being above 1.2 indicates increased production of cytokins that suppress T-lymphocytes in neonatals. The method enables to detect functional defect of immune system that characterizes neonatal period.

EFFECT: higher efficiency of detection.

2 ex

FIELD: medicine.

SUBSTANCE: method involves measuring forced exhalation volume per 1 s (FEV1) in l, full right ventricle evacuation time (RVE) in ms and angiotensin II value (AII) in ng/l. Discriminant relationship is built as D=0.504·RVE+3.038·FEV1 - 2.0·AII. D being less than 83.88, pulmonary hypertension occurrence is predicted within 1 year. D being equal to or greater than 83.88, no pulmonary hypertension is predicted to occur.

EFFECT: enhanced accuracy of prediction.

FIELD: medicine, medicinal immunology.

SUBSTANCE: method involves determination of heterophilic antibodies in human serum blood by the Paul-Bunnel's method relatively the level of circulating immune complexes, complement-activating properties of heterophilic antibodies by incubation of standardized ram erythrocytes with 0.8% serum for 30 ± 5 min and the following measurement of the erythrocytes lysis degree. The measurement of the effector function coefficient of heterophilic antibodies is carried out by the complement system Keff.f.h.a.-c.s. by the formula: Keff.f.h.a.-c.s. = Y/Tg.a. wherein Y means a lysis degree, %; Tg.a. means a reverse titer of heterophilic antibodies to ram erythrocytes. The damage assay is carried out by comparison of the immune status with the relative level of circulating immune complexes in serum. Method provides detection of preclinic from of immunodeficiency and autoimmune diseases that opens the possibility for their prophylaxis at most early stages of development. Invention can be used for assay of damage in the immune status in human serum blood.

EFFECT: improved method for assay.

5 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method involves concurrently examining anti-inflammatory IL-4 level in blood serum and lacrimal fluid. The value being within the limits of 60-70 pg/l in blood serum and 5-15 pg/l in lacrimal fluid, disease prognosis is considered to be unfavorable. The IL-4 concentration being within the limits of 90-100 pg/l in blood serum and 20-30 pg/l in lacrimal fluid, disease prognosis is considered to be favorable.

EFFECT: high accuracy of diagnosis.

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