Method of detection of specific liver disease in leprosy patients

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to leprology, and can be used particularly for the detection of specific liver disease in the leprosy patients. Blood serum of the leprosy patients is analysed to determine the level of US-M.leprae and DIS-BSA leprosy mycobacteria antigen antibodies, and also the concentration of alpha-1-antitrypsin. If observing the level of antibodies to two antigens increased as compared to a norm and the concentration of alpha-1-antitrypsin increased higher than 346.7 mg/dl, liver disease is detected in the leprosy patients.

EFFECT: method provides higher accuracy.

4 ex

 

The invention relates to medicine, namely leprology, and can be, in particular, used to determine the specific liver lesions in patients with leprosy.

From the practice of medicine known method for determination of liver lesions in patients with leprosy, consisting in the fact that in the serum biochemical method determines the activity of gamma-glutamyl transferase (Cemba VP, Naumov old Testament, Yushchenko A.A., Umnov SG, Bezrukavnikov O.A., Urlaeva N. Diagnostic value of the determination of the activity of gamma-glutamyltransferase in serum of leprosy patients with chronic persistent hepatitis // Scientific-practical conference "Topical issues of treatment of sexually transmitted infections, and chronic dermatoses": Abstracts of scientific papers.- Ekaterinburg, 2002. - N.218). The disadvantages of this method is its use only in combination with other diagnostic methods, due to its poor accuracy; increased activity of the enzyme occurs in other diseases (pancreatitis, ischemic heart disease) and conditions (height, prolonged physical activity, pregnancy, circulatory failure), and not always due to chronic liver disease, false-positive test results can be associated with taking patients medicine is the R drugs (anticonvulsants, anticancer, isoniazid) and contribute to overdiagnosis pathological conditions of the liver, which reduces the accuracy of the method. All these shortcomings do not allow to obtain a specific technical result - increase the accuracy of the method.

There is also known a method of determining liver lesions in patients with leprosy, consisting in determining the serum amount of the peroxide-modified Eparistera APO-B-containing lipoproteins. (Patent No. 2188431 from 27.08.2002). The disadvantages of this method is the inability to identify specific lesions of the liver, the complexity and duration of the exercise. In addition, to obtain results, it is necessary to carry out the complex mathematical calculations. These shortcomings do not allow to obtain a specific technical result - increase the accuracy of the method.

The closest method to the present invention is a method of identifying Mycobacterium leprosy patients with regression of the disease, consisting in the detection of antibodies to Mycobacterium leprosy using enzyme immunoassay (Patent No. 2137137 from 10.09.1999).

The similarity of this method to offer is that they both belong to the field of medicine, namely leprology, and consist in the detection of antibodies to Mycobacterium leprosy.

The known method has the following problems is ADI:

- the lack of accuracy of the method due to the fact that the known method it is impossible to determine the localization of specific process;

a well - known method includes the additional definition of cross-reacting antibodies to antigens of Mycobacterium leprosy, which can be found in other diseases;

- the duration of the method due to the necessity of formulating a response counter immunoelectrophoresis;

- accurate localization of specific process according to the autopsy.

These shortcomings do not allow to obtain a specific technical result - increase the accuracy of the method.

Leprosy is a chronic infectious granulomatous disease caused by Mycobacterium leprae (.leprae), characterized by a long incubation period, a prolonged course, a variety of clinical manifestations, prone to periodic exacerbations, affecting mainly the skin, the mucous membranes of the upper respiratory tract, peripheral nervous system, musculoskeletal system and internal organs. Of the last, most often affected is the liver, which usually develops chronic leprosy hepatitis, supported by clinical and laboratory and instrumental methods of examination. Disturbances of liver function in patients with Le is romanosky type of leprosy are expressed in deviation of the protein, pigment, enzyme, carbohydrate metabolism, and also in violation of the antitoxic function of the liver. Leprosy process quite distinct specific and nonspecific changes in the parenchyma and stroma of the organ. In the etiology of liver disease in leprosy cannot be excluded and toxic component caused by the prolonged administration of drugs for a specific leprosy therapy and for the treatment of comorbidity. Thus, the problem of liver disease in leprosy remains one of the most important in modern leprology. Its most important aspect is the timely detection of liver damage, especially since some patients might latent or oligosymptomatic the course of the disease. At the same time, specific liver damage in leprosy is observed at early stages of the disease, and the onset of regression leprosula process is not indicative of intactness liver. In addition, in the presence of functional liver failure increases the probability of adverse effects from leprosy chemotherapy, patients receive for a long time. In this regard, relevant is the search for new ways to determine liver damage.

In clinical practice widely used indicators Belko the acute phase, because of the important role of the latter in the cascade of reactions nonspecific defense in pathological processes.

Along with antibodies able to specifically bind foreign antigens in the blood plasma contains another group of protective protein inhibitors of proteinases. Widely distributed in tissues and biological fluids of proteolytic enzymes having high physiological activity, requires a fine regulation of their actions. In certain pathological conditions caused by inflammatory reaction, excessive activation of proteases that may contribute to the degradation of tissue proteins. An important method of control of proteolysis is the presence in the body specific protein inhibitors of proteolytic enzymes, to the extent necessary to limit their biological activity.

From 70 to 80% antiprotease activity of blood falls on alpha-1-antitripsin. Acting as the main inhibitor of neutrophil elastase and alternative inhibitor entire class of serine proteases, alpha-1-antitripsin natural increases in inflammatory processes. Inhibitors Proteus can be considered as indicators of the severity of the inflammatory response and massive tissue damage, the effectiveness of therapy and prognosis of the disease, and their definition is of great diagnostic value in inflammatory and destructive processes taking place in a number of diseases and conditions, including leprosy.

The chronic nature of the disease, the generalization process, slow regress under the influence of chemotherapy can lead to persistent forms of Mycobacterium leprosy in various organs and tissues - liver, spleen, lymph nodes, nerves. Markers persistence of Mycobacterium leprosy in organs and tissues, leading to the development of specific complications leprosula process are antibodies. Antibody titers reflect the degree of antigenic load, and their determination in serum to evaluate the effectiveness of treatment and may be one of the criteria of cure the sick. At the same time, serological screening of patients with leprosy in the stage regression can identify individuals with the persistence of mycobacteria in tissues and organs and thus to form groups the risk of relapse and the development of specific complications, including liver.

As antigens in serological analysis with leprosy are semisynthetic analogue of specific carbohydrate epitope of phenolic glycolipid-1 of Mycobacterium leprosy and native preparation of Mycobacterium leprosy, t is engaged in a complex of species-specific antigens. Detection in the serum of patients with clinical regression of antibodies to these antigens is indicative of the presence of the bacilli of leprosy in the body.

The present invention solves the main task of improving the accuracy of the method. The essence of the invention is expressed by a set of essential features that are sufficient to provide the technical invention of the result.

The proposed method is that when the detection of antibodies to Mycobacterium leprosy additionally determine alpha-1-antitripsin in the serum and when it is above 346,7 mg/DL judged on specific liver lesions in patients with leprosy.

The proposed method is as follows.

Examined 55 patients with leprosy receiving inpatient and outpatient treatment in FGU "Institute for the study of leprosy University." All patients were divided into groups. The first group consisted of 30 patients who were identified clinical, laboratory and instrumental (according to the results of ultrasound) signs of liver disease of mixed etiology (specific, toxic, viral). The second group included 25 patients who did not have at the time of examination for signs of liver damage.

Alpha 1-antitrypsin in serum define immunoturbidimetric method using commercial is a mini set of the firm "Sentinel", Italy. Measurement is carried out on a programmable photometer "Humalyzer 2000 (Germany). Statistical processing of the obtained results is carried out using student's criterion.

Antibodies to Mycobacterium leprosy is determined as follows. Take blood from a finger using the scarificator in an amount of 0.1 ml in a dry test tube. The tube of blood is placed for 1 h in a thermostat at 37°C, and then in a refrigerator (+4°C) for 1 h for retraction of the clot. Pooled serum is sucked off in a dry test tube and examined using enzyme-linked immunosorbent assay (ELISA). For long-term storage of serum frozen at -20°C. Use two antigenic preparation of M.leprae: native - soluble antigen of M.leprae destroyed by ultrasound, and semi-synthetic (DIS-BSA) on the basis of the carbohydrate epitope of phenolic glycolipid (PGL-1) of M.leprae.

Soluble antigens were obtained from M.leprae isolated from infected tissues of rats with experimental leprosy, according to the method of Draper (Draper P. Problems related to purification of M.leprae from armadillos tissues and standertiration of M.Leprae preparations - Jn.: Report on the Enlarged S.C. Meeting, Geneva, 1979. W.H.O. Document, Annex 1, prat 1/79, p.4-4).

M.leprae allocate by differential centrifugation of tissue homogenates in the density gradient of sucrose, and then separated from the tissue particles in two-phase polymer system consisting of 7% dextran T-500 (Pharmacia) and 5% pole is elangligis with molecular weight of 6000 daltons (BDH company). Peeled .leprae subjected to ultrasonic destruction apparatus M E-100 (UK) in saline solution for 7 min at 18 kHz (kilohertz). Effect of destruction of controlled microscopically. The suspension is destroyed M.Leprae centrifuged (10000 rpm for 40 min d=23 cm), adosados used as the native antigen (USD-M.Leprae). Semi-synthetic antigen (DIS-BSA) were obtained from the Bank who.

Setting ELISA carried out on polystyrene microplate for immunological reactions single use (production Association “metroliner” THE 64-2-375-86).

At the same time sensibiliser two tablets: 1 - USD-M.leprae, 2nd synthetic antigen (DIS-BSA).

The first tablet sensibiliser USD-M.Leprae in carbonate-bicarbonate buffer pH 9,25±0,25 (12,18 g Na2CO3, 3,47 g NaHCO3and 0.2 NaN3dissolve in 1 l distilled water) at a rate of 5 μg of antigen in 1 ml of buffer in a volume of 0.1 ml per 1 well (18 hours at +4°C). The tablet then washed 3 times in Sauternes saline solution and 3 times with distilled water with the addition of 0.05% tween - 20, after which the wells fill in 0.1 ml of 1% solution of bovine serum albumin in 0.02 M phosphate-buffered saline (pH 7.3 to 7.4) and incubated for 1 hour at 37°C. Then re-produce the flushing holes in the above manner and fill in holes on 0.1 ml of the investigated sera ill the x leprosy, diluted to 1:400 in phosphate-saline buffer (pH 7,4)containing 1% calf serum. The tablet also put a positive control serum of patients with active leprosy) and negative control (donor serum at the same dilution (1:400). Each serum sample is poured into 2 wells (duplicate). Tablet incubated at 37°C for 1 hour, and then again repeat the process of washing. Then to each well was added 0.1 ml conjugate (rabbit antibodies to common human immunoglobulins labeled with peroxidase) in a dilution of 1:1000 and incubated 1 hour at 37°C. After wash add substrate mixture of 5-aminosalicylic acid with 0.05% hydrogen peroxide (100 ml) heated to 70°C distilled water, add 80 mg of 5-aminosalicylic acid, the solution is cooled to room temperature and set pH 5,9-6,0; to 18 ml of the resulting solution add 2 ml of 0.05% solution of hydrogen peroxide). Tablet stand at room temperature for 60 minutes

In the presence of serum antibodies to USD-M.leprae substrate mixture is painted in dark brown color (positive reaction), and the control wells remain colorless or slightly yellow (negative reaction). The results of the reaction is evaluated on the photometer (Minireder 590, USA) at a wavelength of 450 nm according to the intensity of the color sample and expressed in units of optical tightly the ti (OP). Positive consider samples, indicators OP in which 2 or more times higher than those in negative control OD. The OD of the negative control does not exceed 0,20.

At the same time carry out sensitization of the second tablets. Antigen DIS-BSA diluted in carbonate-bicarbonate buffer (pH 9,25±0,25) at the rate of 2 μl of the antigen in 1 ml buffer, poured into 0.1 ml per well (1.18 g Na2CO3; 3,74 g NaHCO3and 0.2 NaN3dissolve in 1 l of distilled water) and leave at 37°C for 18 hours. The tablet then washed 3 times in Sauternes saline with 0.05% tween - 20 (34 g NaCl in 4 l of distilled water, bringing the pH to 7.4 with 2 m Na2HPO4). After washing the wells fill in 0.1 ml of 1% solution of bovine serum albumin in 0.02 M phosphate-buffered saline (pH 7.3 to 7.4) and incubated for 1 hour at a temperature of +37°C. Then produce washing as described above and poured into the wells of the test serum (0.1 ml), patients with leprosy, diluted to 1:200 in phosphate-saline buffer (0,584 g NaCl, 12.8 g of Na2HPO4; 2,62 g NaH2PO4containing 1% calf serum. Similarly diluted serum from patients with active leprosy (positive control) and serum donor (negative control). The tablet is incubated for 1 hour at a temperature of +37°C, then washed again. Then to each well add p is 0.1 ml conjugate (rabbit antibodies to human immunoglobulins of class M, peroxidase labeled) at a dilution of 1:1000 and incubated for 1 hour at a temperature of +37°C. After wash add substrate - 5-aminosalicylic acid with 0.05% hydrogen peroxide (100 ml) heated to 70°C distilled water, add 80 mg of 5-aminosalicylic acid, the solution is cooled to room temperature and set pH 5,9-6,0; to 18 ml of the resulting solution add 2 ml of 0.05% solution of hydrogen peroxide). Tablet stand at room temperature for 40-60 minutes

In the presence of serum antibodies to DIS-BSA substrate mixture is painted in dark brown color (positive reaction), and the control wells remain colorless or slightly yellow (negative reaction). The results of the reaction appreciate instrumental in the photometer (Minireader 590, USA) at a wavelength of 450 nm according to the intensity of the color sample and expressed in units of optical density (OD). Positive consider samples, indicators OP in which 2 or more times higher than those of OP in the negative control OD of the negative control does not exceed 0.15.

The results showed that the concentration of alpha-1-antitrypsin deficiency in patients with leprosy in the liver was higher compared to patients with leprosy who do not have this disease, and was 389,77±to 43.1 mg/DL and 298,40±15,68 mg/DL, respectively (p<0,05). Thus, the average value with the holding of alpha-1-antitrypsin (M) in patients with leprosy, with the defeat of the liver (first group)amounted to 389,77 mg/DL. Taking into account the standard error of the mean (t) the lower boundary of the studied parameters in the first group is equal to 346,7 mg/DL. She was adopted as a criterion, when exceeding the values of which in patients with leprosy were defined as damage to the liver.

Serological examination of patients, related to the first and second groups revealed a higher level of antibodies to antigens of Mycobacterium leprosy (USD-M.leprae and DIS-BSA) in patients with signs of liver damage (normal for USD-M.leprae<0,20; for DIS-BSA<0,15). The level of antibodies to both antigens in the second group were in the range of valid values.

Thus, the positive results of serological surveys in conjunction with an increased level of alpha-1-antitrypsin indicate persistence of Mycobacterium leprosy in the liver, as evidenced by clinical, laboratory (blood count) and instrumental (ultrasound) methods.

The proposed method is achieved, in comparison with the prototype, improving the accuracy of the method.

The proposed method was successfully tested in the clinic FGU "Institute for the study of leprosy, Roszdrav" 55 patients during 2007-2009

Below are the results of testing.

Example 1.

Patient M., born in 1932 History no. 1963.

DS: Leprosy-related issues is ichno-lepromatosis type (BL), stage regression.

Clinically observed pain and dyspeptic syndromes, according to the data of laboratory and instrumental methods (biochemical blood analysis, ultrasound examination of abdominal cavity organs): increase the level of aspartate aminotransferase, increased liver size, increased echogenicity and diffusely heterogeneous structure of the body.

The determination of specific lesions of the liver was carried out by the proposed method.

Serological survey on 31.03.2008,

Blood (0.1 ml) was taken in a test tube from a finger was placed in a thermostat (1 h, 37°C), then in a refrigerator (+4°C, 1 h). Selected serum and was titrated in phosphate-buffered saline (pH 7,4)containing 1% calf serum to obtain two dilution: 1:200 and 1:400. In two wells (duplicate) polystyrene tablet, pre-sensitised USD-.leprae (5 μg/ml)were filled with 0.1 ml of serum of the patient M in a dilution of 1:400; two wells were filled with 0.1 ml (1:400) serum pool of donors (negative control), two wells were filled with 0.1 ml (1:400) serum pool of patients with active leprosy (positive control). The tablet was placed in a thermostat (37°C, 1 h), then washed with phosphate-saline buffer (pH 7,4)containing 0.05% tween-20 (FSS T), after which the wells were filled with 0.1 ml conjugate (rabbit antibodies to common human immunoglobulins labeled with peroxidase) is in the attachment 1:1000 and incubated for 1 h at 37°C. Then the tablets were washed FSB-T, the wells were added to 0.1 ml of substrate mixture (5-aminosalicylic acid with 0.05% hydrogen peroxide, pH 5,9-6,0), kept at room temperature for 1 h and was photometrically the results of the reaction. Index OP in the wells with negative control was equal to 0.05. Index OP in the wells with positive control was equal to 0.98. Index OP in the wells with the serum of the patient M was equal to 0.68.

The serum of the patient M in a dilution of 1:200 in the amount of 0.1 ml was poured in two wells (duplicate) tablet, pre-sensitised DIS-BSA. In two wells poured on ML ml serum pool of donors (negative control), two holes poured into 0.1 ml of a serum pool from patients with active leprosy (positive control) in the same dilution (1:200), and the serum of the patient M, and incubated at 37°C, 1 h Then the wells were washed FSB-T (pH of 7.4) was added to 0.1 ml conjugate (rabbit antibodies to human immunoglobulins of class M, peroxidase labeled) at a dilution of 1:1000. Tablet incubated 1 hour at 37°C and then washed as described above. To the wells were added to 0.1 ml of substrate mixture (5-aminosalicylic acid with 0.05% hydrogen peroxide, pH of 5.9 to 6.0). The tablet was kept 35-40 min at room temperature and evaluated photometrically at microphotometer at a wavelength of 450 nm. The measure OD of the negative control b is l equal to 0.03. The measure OD of the positive control was found to be 0.96. Index OP serum of the patient M was equal to 0.29.

Alpha 1-antitrypsin in the serum was determined immunoturbidimetric method using a commercial kit company "Sentinel", Italy. The measurements were performed on a programmable photometer "Humalyzer 2000 (Germany). Statistical processing of the obtained results was performed using student's criterion. The concentration of alpha-1-antitrypsin in serum from 31.03.2008 was 458,1 mg/DL.

Thus, the positivity of the patient together with an increased level of alpha-1-antitrypsin deficiency indicates persistence of Mycobacterium leprosy in the liver, as evidenced by clinical, laboratory (blood count) and instrumental (ultrasound) methods.

Conclusion: the proposed method allows identifying specific liver disease in this patient.

Example 2.

Patient A., born in 1945 History No. 3070.

DS: Leprosy, lepromatous (LLs) type, stage regression.

Clinically noted inconstant pain and dyspeptic syndromes, according to the data of laboratory and instrumental methods (biochemical blood analysis, ultrasound examination of abdominal cavity organs): increase the level of aspartate aminotransferase, elevated levels of total protein, propanganda the content of albumin, the increase in liver size, increased echogenicity and diffusely heterogeneous structure of the body.

The determination of specific lesions of the liver was carried out according to the method described in example 1. Serological study from 2.06.2008 g: antibodies to USD-.leprae - 0,87 (normal <0,20); antibodies to DIS-BSA - 0,19 (normal <0,15). The concentration of alpha-1-antitrypsin in serum from 2.06.2008 hastavra 431,4 mg/DL.

Thus, the positivity of the patient together with an increased level of alpha-1-antitrypsin deficiency indicates persistence of Mycobacterium leprosy in the liver, as evidenced by clinical, laboratory (blood count) and instrumental (ultrasound) methods.

Conclusion: the proposed method allows identifying specific liver disease in a particular patient.

Example 3.

Patient N., born in 1932 History No. 3645.

DS: Leprosy, lepromatous (LLs) type, stage regression.

Clinically and information about liver disease are absent. According to ultrasound: liver enlarged, echo is increased, the structure of the diffuse inhomogeneous.

The determination of specific lesions of the liver was carried out according to the method described in example 1. Serological study from 4.02.2008 g: antibodies to USD-.leprae - 0,11 (normal <0,20); antibodies to DIS-BSA - 0,16 (normal <0,15). The concentration of alpha-1-antitrypsin in serum from 4.02.208, was 314,0 mg/DL.

Conclusion: slight increase in levels of antibodies to only one of the two antigens (DIS-BSA) together with the concentration of alpha-1-antitrypsin deficiency below a set criterion does not allow to reliably determine the specific liver disease in a particular patient.

Example 4.

Patient K., born in 1934 History No. 3829.

DS: Leprosy, lepromatous type (LLs), stage regression.

Clinical and laboratory signs of infection are absent. The determination of specific lesions of the liver was carried out according to the method described in example 1. Serological study from 26.05.2008 g: antibodies to USD-M.leprae - 0,04 (normal <0,20); antibodies to DIS-BSA - 0,03 (normal <0,15). The concentration of alpha-1-antitrypsin in serum from 26.05.2008 was 132,0 mg/DL.

Conclusion: the obtained results indicate the absence of specific lesions of the liver.

The proposed method is achieved, in comparison with the prototype, improving the accuracy of determination of specific liver lesions in patients with leprosy. The proposed method allows to accurately determine not only the localization of the pathological process, but also in vivo to establish etiology. In addition, the proposed method early, preclinical, identifying specific signs of liver damage in patients with leprosy in the active stage,and the stage regression. The proposed method does not require the reaction of the counter immunoelectrophoresis and complex mathematical calculations, which significantly reduces the time of its implementation, there is no need for further definition of cross-reacting antibodies to antigens of Mycobacterium leprosy, which can be found in other diseases. The method can be reproduced in the clinical laboratory, is minimally invasive and burdensome for the patient.

Thus, the authors proposed earlier by anyone not offer a way to define specific liver lesions in patients with leprosy.

The proposed method can be recommended for use in leprosy and STI institutions of the country's health system.

Method for determination of liver lesions in patients with leprosy, including the study of blood serum, characterized in that to determine the serum level of antibodies to antigens of Mycobacterium leprosy USD-M.leprae and DIS-BSA and alpha-1-antitrypsin, and when the level of antibodies to the two antigens relative to the standards and content of alpha-1-antitrypsin above 346,7 mg/DL determine liver damage in patients with leprosy.



 

Same patents:

FIELD: medicine.

SUBSTANCE: essence of invention includes selection of sample for testing from area of malignant affection in patient, suffering from cancer of large intestine, selection of control sample, measurement of the level of one or several genetic markers, selected from the group, comparison of measured levels in experimental samples with control ones. Reducing of levels of one or several genetic markers in comparison with control sample, indicates increased resistance to docetaxel. Also described are sets which allow to predict or realise monitoring of patient's response to docetaxel.

EFFECT: identification of genetic markers, by which it is possible to realise patient's response to chemical therapy by docetaxel.

18 cl, 9 ex, 3 tbl, 16 dwg

FIELD: chemistry; biochemistry.

SUBSTANCE: present invention relates to molecular biology and can be used in designing agent and methods of modulating body functions associated with HGF/c-met signalling pathway. The invention discloses HGF/c-met polypeptide-antagonists which are mutant forms of HGF which contain a mutation in the N-terminal part of the β-chain and/or in its dimerisation part. The disclosed polypeptides have lower biological activity compared to wild type HGG and can be used in modulating activity of c-met, cell proliferation, cell migration and angiogenic cell activity.

EFFECT: invention describes a method of obtaining HGF muteins using DNA recombinant technology and agents which are necessary for its existence.

22 cl, 8 dwg, 1 ex

FIELD: medicine.

SUBSTANCE: in order to predict insufficiency of saturation with oxygen of peripheral blood of pregnant women who have herpes-virus infection, content of glucose-6-phosphat-dehydrogenase and TNFα in peripheral blood is determined. Discriminant equation D=-0.589·G-6-PDG+{+1.23·TNFα} is solved. If value of discriminant function is equal or greater than 89.95, predicted is development of oxygen insufficiency, accompanied with reduction in peripheral blood of pregnant women of pO2 with titre of antibodies to Herpes simplex virus 1:12800 to 27.63±0.7 mm Hg, and of HbO2 to 90.2±0.47% (control: pO2 - 39.22±0.5 mm Hg; HbO2 - 95.3±0.27%).

EFFECT: increased accuracy of determining possibility of development of oxygen insufficiency in pregnant woman with herpes-virus infection.

3 tbl

FIELD: medicine.

SUBSTANCE: in estimation of local inflammation activity in case of background diseases of neck of uterus sampling of analysed biomaterial is performed by bringing of device for native material removal to external neck of uterus fauces. Said device has elongated holder in form of rod with length, exceeding length of vagina. From distal side of rod, at angle to it, located is operation element in form of roller. Proximal end of rod serves as manipulator-handle. Sampled material is placed until complete submergence into tightly closable test tube, filled with 1.0 ml of 0.155 M solution of sodium chloride. Mixture is centrifuged for 10 minutes. Obtained materials are analysed by method of solid phase ELISA analysis on boards by means of set of reagents "Vector-Best". Concentration of cytokine of interleukin-6 in analysed samples is determined spectrophotometrically. If value is higher than 50 pg/ml to 92 pg/ml, conclusion about presence of chronic inflammation process is made. If value is higher than 92 pg/ml, conclusion about active course of inflammatory process and expressed immune response is made.

EFFECT: application of method allows to increase quality of analysed native material and accuracy of obtained result without complicating analysis process.

2 cl, 3 ex

FIELD: medicine.

SUBSTANCE: invention can be used for diagnostics of presence of destructive process in uterine appendages in case of localised peritonitis if gynecological genesis, and for selection of optimal treatment tactics. In order to realise the method content of C-reactive protein (CRP) in blood serum is determined by turbodimetric method, if values of C-reactive protein content are 130 mg/l and higher, purulent-destructive process in uterine appendages in case of localised peritonitis is diagnosed, if values of C-reactive protein content are lower than 130 mg/l diagnosed is purulent salpingitis in case of pelvioperitonitis.

EFFECT: application of method allows to detect presence of purulent-destructive process in uterine appendages in case of pelvioperitonitis in urgent way, which determines tactics of further treatment.

2 ex

FIELD: medicine.

SUBSTANCE: invention can be used for obtaining diagnostic and/or therapeutic agents, in particular antibodies, which specifically interact with proteins of cell surface of target cells. Claimed is method of screening phage display library and including it method of isolation of library element, which is specific partner of binding of target cell surface protein, and method of isolation of unknown cell surface protein, specifically interacting with library element. Method of screening phage display library according to invention includes contact of said library with cells of interest with further separation of cells, which bound with one or several elements of expression library, from non-bound elements by separation through organic phase, and differs by introduction of additional stage, which consists in carrying out before said separation of at least one stage of material washing, and ensures essential increase of method efficiency.

EFFECT: higher efficiency.

30 cl, 7 dwg, 6 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to ophthalmology, can be used for prediction of progression of myopia acquired in schools in 10-14 and 15-17 year old children. Blood serum is analysed for cartilage glycoprotein-39 with the use of sandwich-type enzyme immunoassay. If its concentration is within 34.7-53.4 ng/ml in 10-14-year-old children and 22-56.3 ng/ml in 15-17-year-old children, a permanent course of myopia is predicted. The concentration 18.5-33.9 ng/ml in 10-14-year-old children and 53.7-69 ng/ml in 15-17-year-old children enables to predict a progressive course of myopia.

EFFECT: use of the invention improves accuracy of the prediction of clinical course.

4 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to diagnostic techniques and concerns a diagnostic techniques technique for respiratory function of peripheral blood erythrocytes in pregnant women in herpes virus infection episode in the third trimester. The technique consists in determination of glutathione reductase and herpes virus antibody titre concentration by the enzyme immunoassay in peripheral blood in pregnant women suffered from a herpes virus infection episode. If glutathione reductase concentration is 8.36±0.137 E/g Hb (ref. - 7.82±0.185 E/g Hb), herpes virus antibody titre is 1:6400. If herpes virus antibody titre is increased to 1:12800, erythrocyte glutathione reductase concentration is decreased 4.48±0.22 E/g Hb.

EFFECT: technique is characterised by high sensitivity and allows predicting the development of impaired respiratory activity of erythrocytes in the pregnant women with the herpes virus infection episode.

FIELD: medicine.

SUBSTANCE: simultaneously two parametres are determined: for men - in ejaculate, and for women - in discharge of cervical canal, namely, slgA and lactoferrin. If level of secretory immunoglobulin A is 3.01±0.50 mcg/ml and lower and simultaneous level of lactoferrin is 6876±89 ng/ml and higher, gonococcus infection with systemic manifestations is diagnosed, if said condition is absent, localised gonorrhea is diagnosed.

EFFECT: application of claimed method allows carrying out differential diagnostics of various forms of gonococcus infection in case of its oligosymptomatic course.

3 ex, 2 tbl

FIELD: medicine.

SUBSTANCE: before and after treating chronic heart disease, modified LP(a) are determined as follows. Blood serum 0.6 ml is treated with 0.1% Triton X-100 0. 2 ml, incubated for 15 minutes at 20°C. Then 7% polyethylene glycol 6000 is added, incubated with the Sudan Black stain at 40°C for 1 h; it is followed with electrophoretic separation of LP in agarose gel in a cavity 4×20 mm. The decreasing level of modified LP(a) by 40% and more in comparison with the primitive level, treating chronic heart disease treatment is considered as effective.

EFFECT: higher accuracy of estimating clinical effectiveness of chronic heart disease.

1 tbl, 6 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to gastroenterology and cardiology, can be used in medical institutions for determination of severity level of metabolic disorders and risk of metabolic syndrome in the patients suffering chronic cholecystitis. For risk evaluation of metabolic syndrome in the patients with chronic cholecystitis by the severity of immune-metabolic disorders, the following significant diagnostic characters are used: fasting glucose, apoprotein atherogenicity coefficient (apoB/anoAl), high density lipoprotein cholesterol, T-helpers and T-suppressors ratio, phagocytic index. The derived values are entered in an arithmetic formula to evaluate the risk of metabolic syndrome by the severity level of immune-metabolic disorders by total value expressed in standard units.

EFFECT: method facilitates work of a doctor, allows to evaluate the patient's status in a relatively short time, to predict the risk of metabolic syndrome and to prescribe the well-timed therapy to prevent the progression of metabolic syndrome and cardiovascular complications in the patients with chronic cholecystitis.

1 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to neurosurgery. It involves clinico-neurological examination to be carried on. A degree of nerve conduction abnormality is determined. Neurofibroma is diagnosed by the presence of pain symptom, a palpated space-occupying lesion and the absence or functionally insignificant decrease in nerve trunk conductivity. Neurinoma is diagnosed by the presence of pain symptom, a palpated space-occupying lesion and the evident functionally significant abnormalities of nerve trunk conductivity.

EFFECT: technique provides a noninvasive procedure, higher diagnostic accuracy, the absence of tumour malignisation, and reduction of recurrences.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to oncology and can be used for prediction of cerebral metastases of pulmonary cancer The method involves the determination of prognostic indices by clinical anamnestic data If observing such prognostic indices as 55-60-year-old age in males, diagnosed peripheral adenocarcinoma or small cell carcinoma of right lung, T2-T3N2 (IIIA) stage, non-administered adjuvant therapy, the occurrence of cerebral metastases, mainly in the left brain is predicted within approximately 10 months after a primary tumour has been detected.

EFFECT: use of the invention allows individually predicting the probability and time of manifestation, localisation of cerebral metastases

1 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to ophthalmology, can be used for prediction of progression of myopia acquired in schools in 10-14 and 15-17 year old children. Blood serum is analysed for cartilage glycoprotein-39 with the use of sandwich-type enzyme immunoassay. If its concentration is within 34.7-53.4 ng/ml in 10-14-year-old children and 22-56.3 ng/ml in 15-17-year-old children, a permanent course of myopia is predicted. The concentration 18.5-33.9 ng/ml in 10-14-year-old children and 53.7-69 ng/ml in 15-17-year-old children enables to predict a progressive course of myopia.

EFFECT: use of the invention improves accuracy of the prediction of clinical course.

4 ex

FIELD: medicine.

SUBSTANCE: method relates to medicine, hepatology. In a patient with chronic viral hepatitis, abdominal ultrasound is applied. The following diagnostic criteria are observed: thickness of left lobe of liver; length of spleen, liver size, liver echogenicity, diametre of splenic vein; diametre of superior mesenteric vein, diametre of hepatic artery, mean blood velocity in portal vein. The histology activity index is calculated by an original mathematical formula.

EFFECT: method enables a noninvasive morphological examination of hepatic tissue sampling to determine the histology activity index in patients with chronic viral hepatitis.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to aggressive medical therapy, resuscitation science, critical care medicine, laboratory diagnostics and can be used by resuscitators, intensivists, laboratory doctors for well-timed diagnosis and consequently, for individualised aggressive medical therapy of acute disseminated intravascular coagulation. The integrated assessment of links of haemostatic system and the clinical appraisal of organ dysfunction are applied in a measurer, and when observing structural hypercoagulation characterised by fibrinogen level increase, thrombocyte activity increase, growth of soluble fibrin complex (SFC) level, and also when observing chronometric hypercoagulation characterised by time tests, palette-derived factor 4 activity (P4) with manifested petechial haemorrhage and organ dysfunction, and coagulation cascade activation with underlying depression of antithrombin III and protein C, a hypercoagulation stage of acute DIC is diagnosed. Chronometric hypercoagulation by Activated partial thromboplastin time (APTT), INR, fibrinogen and P4 with manifested signs of structural hypocoagulation by thrombin time prolongation and D-dimer activity increase with underlying further intensification of anticoagulant system deficiency, progression of target organs dysfunction and mixed haemorrhage show a transitive stage of acute DIC. If observing said hypocoagulation changes and disturbed fibrinolytic activity with prevailing either decompensated organ and tissue dysfunction, or hemorrhagic syndrome up to system haemorrhages, or their combination with hemorrhagic syndrome characterised by polymorphism of clinical picture and localisation: petechial-haematoma haemorrhage at the stress-induced stomach ulcers, hematuria, a coagulopathy stage of acute DIC is diagnosed that is characterised either by depression of fibrinolysis, or preserved fibrinolytic activity, or by activation of secondary fibrinolysis, or by acute primary fibrinolysis.

EFFECT: method allows optimising classification of acute disseminated intravascular coagulation, improving diagnostic significance of the classification and simplifying a diagnostic prospecting which provide a basis to consider the staging of acute disseminated intravascular coagulation.

1 dwg, 4 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to diagnostic techniques and concerns a diagnostic techniques technique for respiratory function of peripheral blood erythrocytes in pregnant women in herpes virus infection episode in the third trimester. The technique consists in determination of glutathione reductase and herpes virus antibody titre concentration by the enzyme immunoassay in peripheral blood in pregnant women suffered from a herpes virus infection episode. If glutathione reductase concentration is 8.36±0.137 E/g Hb (ref. - 7.82±0.185 E/g Hb), herpes virus antibody titre is 1:6400. If herpes virus antibody titre is increased to 1:12800, erythrocyte glutathione reductase concentration is decreased 4.48±0.22 E/g Hb.

EFFECT: technique is characterised by high sensitivity and allows predicting the development of impaired respiratory activity of erythrocytes in the pregnant women with the herpes virus infection episode.

FIELD: medicine.

SUBSTANCE: method of chewing system functioning evaluation refers to medicine, particularly to orthopaedic dentistry. A chewing test involves face bow recording in patients. A face bow index (FBI) is calculated by formula: where vtµ is a wave rate with the most frequent chewing wave length in a chewing phase, vhµ is a wave rate of the same amplitude in a chewing phase, t is a duration of a chewing phase actually (s), v is a number of masticatory movements in a chewing phase, Δt is a mean difference of the length of the most frequent wave and the other chewing waves (s). If the index is 2 and more, chewing system functioning is considered to be good. If the index is less than 2, lowered system effectiveness is observed.

EFFECT: application of the method aims at the objectification of human chewing system functioning evaluation when refining a diagnosis, allows evaluating chewing system functioning, its efficiency, proving the necessity and nature of remedial rehabilitation actions.

2 ex, 4 dwg, 1 tbl

FIELD: medicine.

SUBSTANCE: differential diagnostics of herpetic infections of gastric mucosa us membranes of a stomach or dental pulp is ensured by coordination marking of a biopsy material. Histologic sections of thickness no more than 3 mcm are prepared. The prepared sections are applied on slides as follows: the first section is arranged on the first slide, the second section is applied on the second, the third slide is used for the third section, the fourth section is placed on the first slide and further the same sequence, the number of slides is determined by the identified virus count. Each preparation is analysed for specific viruses of the herpes groups by immunohistochemistry with using specific monoclonal antibodies. The results of each reaction are recorded by the presence of specifically stained antigen granules in a histologic sample, and the images of the results of each reaction are saved in the electronic form. The formed images of the following sections are aligned by the coordination mark. If observing the specifically stained antigen granules of various localisation, herpetic mixed infection of gastric mucosa and dental pulp is diagnosed. When observing the specifically stained antigen granules of the same localisation herpetic monoinfection with immune cross-reaction is diagnosed.

EFFECT: technique presents higher accuracy of diagnosing herpetic infections of gastric mucosa or dental pulp ensured by differential diagnostics between mixed infection and monoinfection with immune cross-reaction.

3 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: total number of slow muscle fibres (NSMF), total number of fast muscle fibres (NFMF), total number of fast muscle fibres type IIA (NFMFIIA) and total number of fast muscle fibres type IIB (NFMFIIB) are determined; genetically total factor F is calculated by formula: F=NFMF/NSMF), while total body factor of FMF type IIA - FIIA and total body factor of FMF type IIB - FIIB are evaluated by formulae: FIIA=NFMFIIA/NSMF, FIIB=NFMFIIB/NSMF. If F exceeds 3 and the FIIA to FIIB relation exceeds 3, a strong unbalanced active type of the HNA is observed; if F exceeds 3, and the FIIA to FIIB relation is equal to 1 to 3, a strong balanced mobile type of HNA is determined; if F exceeds 3, and the FIIA to FIIB relation is equal to 0.3 to 1, a strong balanced inert type of the HNA is present. If F exceeds 3, and the FIIA to FIIB relation is less than 0.3, a strong unbalanced passive type of the HNA is diagnosed; if F is within 0.3 to 3, and the FIIA to FIIB relation exceeds 3, a moderate unbalanced active type of the HNA is observed; if F is within 0.3 to 3, and the FIIA to FIIB relation is estimated within 1 to 3, a moderate balanced mobile type of the HNA is evident; if F is within 0.3 to 3, and the FIIA to FIIB relation is within 1 to 0.3, a moderate balanced inert type of the HNA is diagnosed; if F is within 0.3 to 3, and the FIIA to FIIB relation is less than 0.3, a moderate unbalanced passive type of the HNA is present. If F is less than 0.3, and the FIIA to FIIB relation exceeds 3, a weak unbalanced active type of the HNA; if F is less than 0.3, and the FIIA to FIIB relation is within 1 to 3, a weak balanced mobile type of the HNA is indicated; if F is less than 0.3, and the FIIA to FIIB relation is within 0.3 to 1, a weak balanced inert type of the HNA is indicated; if F and the FIIA to FIIB relation is less than 0.3, a weak unbalanced passive type of the HNA is shown.

EFFECT: method extends the range of products for type determination.

3 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

Up!