Method of diagnosing kidney allotransplant rejection

FIELD: medicine.

SUBSTANCE: method of diagnosing rejection of kidney allotransplant includes determination of phase height PH of peripheral blood live lymphocytes by method of phase-interference microscopy, determination of quantity of lymphocytes with phase height PH≤1.5 mcm, 1.5 mcm<PH≤2 mcm, 2 mcm<PH≤2.5 mcm, PH>2.5 mcm, selection of lymphocyte activity coefficients for each limit, said phase heights of lymphocytes equal k3=3, k2=2, k1=1, k0=0 respectively. Obtained data are used to determine functional activity of lymphocytes in sample by formula: FA=(k3n3+k2n2+k1n1+k0n0)/n, where n is number of lymphocytes in sample, n3 is number of lymphocytes with PH≤1.5 mcm, n2 is number of lymphocytes with 1.5 mcm<PH≤2 mcm, n1 is number of lymphocytes with 2 mcm<PH≤2.5 mcm, n0 is number of lymphocytes with PH>2.5 mcm, k3, k2, k1, k0 are coefficients of lymphocyte activity, and if value of lymphocyte functional activity is within FA=1.8-2.0, rejection of kidney transplant is diagnosed.

EFFECT: application of claimed method makes it possible to set diagnosis in due time, which considerably increases efficiency of anti-crisis therapy in post-transplantation period.

2 ex, 1 tbl

 

The invention relates to medicine, in particular for laboratory research in transplantation.

Allotransplantation kidney is the method of choice in treatment of patients with end-stage chronic renal failure (CRF)which supported the life of the patient, is its social and medical rehabilitation.

An important factor in the deterioration of the function of the renal allograft is acute rejection in the early postoperative period, therefore, timely diagnosis and adequate treatment are important predictors of a favorable clinical outcome.

Rejection - specific immune reaction to transplantation antigens of the donor, causing an inflammatory lesion of the graft. The main morphological components of this process are tubulo-interstitial nephritis is a cellular type of exclusion and Takayasu in the vascular type of rejection, a rejection reaction involved almost all the mechanisms of cellular and humoral response. Despite the use of modern immunosuppressive drugs, acute rejection is a frequent cause of graft loss.

Currently, the diagnosis of acute rejection is based on a comprehensive assessment of clinical symptoms and signs, ISM is on the performance of laboratory and special instrumental methods. Monitoring the development of acute rejection reactions performed on the basis of clinical, instrumental and laboratory and immunological data.

A known method for the diagnosis of transplant rejection (the"Transplantation". Under the editorship of academician V.I. Shumakov. M: "MIA", 2006. - S-506) - percutaneous biopsy of the renal allograft, which allows you to conduct morphological analysis of the pathological processes in ATP using histological, immunological and electron microscopic studies. Percutaneous biopsy of renal transplant is performed by the surgeon in aseptic conditions using a special biopsy needle under control of the ultrasonic sensor, followed by fixing, painting and cooking micropreparative.

The disadvantages of this method are the invasiveness of the possibility of developing complications, the involvement of several experts, the use of expensive reagents for preparation of microscopic specimens.

A known method for the diagnosis of transplant rejection (the"Transplantation". Under the editorship of academician V.I. Shumakov. M: "MIA", 2006. - S), based on the definition of complex laboratory indices of peripheral blood and urine of the recipient. Laboratory signs of rejection are as follows: in the blood - the sharp increase in creatinine (25% from the original is about) and urea, leukocytosis, lymphocytosis, a sharp increase in sedimentation rate, hyperkalemia, hypercoagulation; in urine - increased proteinuria and decrease the specific gravity of the urine. Research is carried out by standard techniques on a daily basis.

In addition to acute rejection reactions, these symptoms can occur in different combinations in infectious complications, surgical complications, toxicity of calcineurin inhibitors and other drugs.

The disadvantages of this method: the absence of strict specificity for acute rejection reactions, it is impossible to diagnose rejection reactions in the subclinical stage, rejection reactions without clinical dysfunction of the graft.

The closest is a method for diagnosing rejection, kidney transplant, including the determination of the phase height PH of living of peripheral blood lymphocytes using phase-interference microscopy, the number of lymphocytes in the sample (RF patent for the invention №2348932, IPC G01N 33/48, publ. 2009). The method is based on the evaluation of morphological and functional parameters of living unfixed and unstained peripheral blood lymphocytes of renal transplant recipients. To achieve a technical result carry out comprehensive rapid assessment linear (geometric diameter, perimeter, area is d') and space-structural (refractive - height and volume) metrics of each lymphocyte using laser interference microscope, allowing to study living cells in transmitted light. Diagnosis is based on the evaluation of phase portraits of lymphocytes. Determine the value of the heterogeneity of the nuclear structures of living cells and when the index value of less than 0.2 to not less than 20% of the cells, make a conclusion about the beginning of the rejection of renal transplant.

The disadvantages of the method are its high complexity and lack of precision, because the use to assess the status of lymphocytes as geometric and phase parameters does not allow to take into account the constancy of the quantity of the nuclear substance cells because the activation of small blood lymphocytes can be misleading due to the use in the calculation of the diameter. Difficult the introduction of the rate of activity of lymphocytes in clinical practice, because it requires two stages of mathematical analysis data.

The task of the invention is to remedy these disadvantages due to the fact that only considers the phase parameters of the nuclear structures of peripheral blood lymphocytes that most accurately describes the processes of activation of lymphocytes in rejection of the renal allograft.

For this purpose, in a method of diagnosis from the Argania kidney transplant, including the definition phase height PH of living of peripheral blood lymphocytes using phase-interference microscopy, the number of lymphocytes in the sample was asked to identify the number of lymphocytes with phase height PH ≤ 1.5 µm, 1.5 µm ≤ PH ≤ 2 µm 2 µm ≤ PH ≤ 2.5 μm, PH ≥ 2,5. New is also the fact that choosing the coefficients of lymphocyte activity for each limit specified phase heights lymphocytes equal to k3=3, k2=2, k1=1, k0=0, respectively, the data and determine the functional activity of lymphocytes in the sample FA by the formula:

FA=(k3n3+k2n2+k1n1+k0n0)/n

where n is the number of lymphocytes in the sample,

n3the number of lymphocytes with PH ≤ 1.5 µm,

n2the number of lymphocytes with 1.5 µm ≤ PH ≤ 2 µm,

n1- the number of cells with 2 µm ≤ PH ≤ 2.5 μm,

n0the number of lymphocytes with PH ≥ 2.5 and

k3, k2, k1, k0- the activity coefficients of the lymphocytes

and when the value of the functional activity of lymphocytes within FA=1,8-2,0 diagnose rejection kidney transplant.

The selected indicator diagnosis of FA, which determine the rejection kidney transplant, characterizes the refractive index for all components of living blood lymphocytes and depends on the nature of the intracellular substances is STV, its concentration and conformational state.

The main volume of the lymphocyte is the nucleus, and nuclear proteins contribute to the optical parameters of the cell, such as a phase height. The number of chromatin in somatic cell standard (diploid) set of chromosomes is constant and does not depend on the majority of factors practically does not change the contents of histone and nonhistone proteins, suggesting that a constant concentration of nuclear matter. The refractive index for all components of cellular structures average of 1,088 and may vary in a small range when the change in the conformational state of the molecules is less than the dimension of the complex of proteins, DNA and RNA, the lower the refractive index. Thus, the phase height of lymphocytes depends primarily on the degree of packing of the chromatin in the nucleus and activity of the processes of protein synthesis. That is, this value allows you to indirectly assess the process of rejection of the renal allograft. For the phase height of lymphocytes there is an inverse dependence on the degree of activation is less than the PH value, the higher the activity of the cell. A practical way limits were established phase heights for different levels of activation. The most active are cells with a PH of less than 1.5 μm, which corresponds to the maximum degree of decondensation the HRO is Atina and activation of lymphocytes, this group was assigned the activity coefficient equal to 3. Similarly identified the activity coefficients for the other phase within the heights.

The results of the study found that the number of lymphocytes based on their activity coefficient in relation to the total number of lymphocytes in the sample determine the amount of activity of the entire population.

The method is as follows.

Lymphocytes were isolated from peripheral blood (PC), stabilized by heparin at a concentration of 25 IU per 1 ml of a standard method of Vosp on the density gradient for lymphocytes Ficoll-Paque (p=1,077 g/cm3) manufactured by Amersham Biosciences (Sweden). The blood was diluted with saline 1:1, then PC has layered in the centrifuge tube on a layer of the gradient. Centrifuged at 1500 rpm for 45 minutes Ring mononuclear cells at the border of the gradient was collected and washed with saline solution twice for 5-7 min at 1000 rpm

The cells were placed in the Goryayev camera with a mirror-coated and examined using the FMC Cytoscan. The sample for one study was 50-100 cells. The phase portraits of lymphocytes were processed using mathematical package MatLab.

As a criterion for the diagnosis of rejection was defined functional activity of the sample of lymphocytes FA, as the magnitude of the back of proportional the th phase a height PH of each cell in the sample, characterizing conformational state nucleolar proteins and the degree of decondensation chromatin: FA=(k3n3+k2n2+k1n1+k0n0)/n, where FA is the functional activity of lymphocytes; n is the number of cells in the sample, n3the number of lymphocytes with PH ≤ 1.5 µm, n2the number of cells with 1.5 µm ≤ PH ≤ 2 µm, n1the number of cells with 2 µm ≤ PH ≤ 2.5 μm, n0the number of cells with PH ≥ 2,5, k3, k2, k1, k0factor activity of lymphocytes.

Research was conducted nuclear structures lymphocytes by the method of computing the phase morphometry in healthy persons In this group included 30 people were studied over 2000 phase-interference portraits with the definition of the functional activity of the nuclei vital unstained peripheral blood lymphocytes:

FA=(3·100+2·196+89+0·41)/426=1,83 for T-lymphocytes;

FA=(3·109+2·19,8+90+0·42)/439=1,85 for b-lymphocytes.

In healthy individuals is FA amounted to 1.83±0.1 T and 1.85±0.1 b-lymphocytes. Received high scores are associated with a predominance of lymphocytes with medium and low values of the phase height, i.e. functionally active cell.

Examined 18 patients with end-stage chronic renal receiving renal replacement therapy, which subsequently executed kidney transplantation. Dann the group represented 50% of women (n=9) and 50% of men (n=9), the average age was 42±9.5 years.

The reasons that led to end-stage chronic renal failure were chronic glomerulonephritis - 61,1% (n=11), chronic pyelonephritis is 22.2% (n=4), chronic tubulo-interstitial nephritis and 5.6% (n=1), syndrome Alport and 5.6% (n=1) and gout 5,6% (n=1).

Method of renal replacement therapy in renal transplant recipients in this group was peritoneal dialysis in 39% (n=7), hemodialysis 61% (n=11). Average time on dialysis amounted to 29.7±18,4 months.

This group was also determined for the functional activity of the nuclei of lymphocytes:

FA=(3·15+2·210+1·102+0·36)/363=1,56 for T-lymphocytes;

FA=(3·12+2·192+1·99+0·33)/336=1,54 for b-lymphocytes.

In table 3.1.1. presents the values of the functional activity of the nuclei of T - and b-lymphocytes of patients THEN undergoing dialysis and healthy individuals (donors).

Table 3.1.1
FA T - and b-lymphocytes of patients DPN (M±σ)
Group inspectionThe value of FA T-lymphocyteThe value of FA b-lymphocytes
Donors N=301,83±0,11,85±0,1
Patients DPN N=181,56±0,15*1,54±0,2*
Note: the significance of differences p<0,05; *differs from control

As for T-and b-lymphocytes is characterized by a significant decrease in the functional activity of the nuclei of cells. On the basis of the obtained results we can assume that in T - and b-immunity there are signs of functional disability associated, apparently, with the suppression of cell activation. At the same time, the data obtained do not allow to assess the proliferative capacity of lymphocytes, but only characterize the status of immunoreactivity in patients with end-stage CRF.

After kidney transplantation research was conducted nuclear structures lymphocytes by the method of computing the phase morphometrics in the early postoperative period.

11 patients were observed during uncomplicated with the primary function of the graft. The measure of the functional activity of the nucleus was 1.4±0,27 as for T-and b-lymphocytes.

In 6 patients with delayed from nitrogen and vadovedyelare function without transplant immunological conflict (no infectious complications) FA was 1.43±0.3 for all cell types.

Thus, there were no significant differences in functional the activity of the nuclei in patients with primary and delayed graft function.

In 3 patients with ORO to the manifestation of clinical manifestations is the functional activity of the nucleus was increased to 1.9±0.1 T and/or b-cells depending on the type of reaction.

After conducting protivogradovoy therapy "heartbeats" metilprednisolona up to a total dose of 1.5-3.0 g and antithymocyte globulin - ATG (Fresenius), FA was comparable with the rate in the group with uncomplicated.

Example 1.

Patient B., born in 1973, with a diagnosis of chronic glomerulonephritis, chronic renal failure, end-stage, arterial hypertension, myocardial observed in MONICA 2005

Considers himself ill for more than 10 years, when a survey revealed high values of HELL, changes in the urine. The deterioration since 2003, when a marked increase in creatinine, urea, proteinuria. Conducted conservative treatment with prednisolone. Since 2005 ESRD, end-stage. Start of treatment peritoneal dialysis since March 2005, with the use of low and sredneoblastnyh solutions. In 2006, he suffered 2 episodes dialysis peritonitis.

14.10.07 performed allotransplantation cadaveric kidneys with the coincidence of the two antigens from the donor 40 years, died of a head injury. Time primary heat ischemia 25 minutes, expiration conservation amounted to 30 hours. Immunosuppressive therapy consisted of prograf, prednisone, metypred,CellCept, zenapax. Function delayed graft, continued treatment with peritoneal dialysis. Diuresis on 1, 3, 5, 7, 9, 11 day amounted to 400, 270, 250, 230, 330, 350 ml, respectively. The level of plasma creatinine 1, 3, 5, 7, 9, 11 day 1,24; 1,3; 1,0; 1,03; 0,89; 0,8 mmol/l, respectively.

According to the complex laboratory of peripheral blood and urine recipient was administered computer. 5 and 11 days after allotransplantation was performed phase morphometry T - and b-lymphocytes of peripheral blood. Sampling populations of each type of lymphocytes ranged from 70 to 100 cells. Was determined by the height of each phase of the lymphocyte and the value of this indicator cell was assigned the corresponding activity coefficient from 0 to 3. Counts the total number of lymphocytes and the number of cells in each group and calculated the functional activity of T-lymphocytes:

for 5 days FA=(k3·8+k2·50+k1·20+k0·2)/80=(3·8+2·50+1·20+0·2)/80=1,86;

for 11 days FA=(3·6+2·20+1·36+0·38)/100=0,94;

and b-lymphocytes:

for 5 days FA=(3·10+2·44+1·18+0·4)/76=1,79;

for 11 days FA=(3·16+2·50+1·14+0·12)/92=1,76.

FA b-lymphocytes despite standard post-transplant drug therapy remained high, allowing us to assume the presence of rejection reactions in the vascular type.

24.10.07, (10 day) performed a biopsy of the kidney transplant.

Geest is the logical study of biopsy kidney.

Light microscopy performed with dyed - hematoxylin-eosin, CHIC-reaction, trigram on Mason. In preparation 13 glomeruli, there is a small delay cells in the lumen of the capillary loops in the rest of the glomeruli without features. Swelling of interstice. Diffuse rare infiltration by mononuclear cells without marked tendency to invasion in tubules (tubelite 4-5 lymphocytes at the edge of the tubule). Tubules look enhanced by reducing the height of the tubular epithelium. 1 middle-sized arteries are individual cells in the subendothelial region. In small arteries and arterioles pronounced vacuolization of myocytes.

Results immunofluorescence: IgG - no, IgA - no, IgM - no, C3 - no, fibrin - no, C4D on the vascular endothelium is defined.

Conclusion: acute vascular rejection 2A (according to the classification of Banff), residual effects of acute tubular necrosis.

On the 8th day started antichrista therapy ATG in a total dose of 500 mg.

After conducting antikrizove therapy functional activity of lymphocytes decreased:

T-lymphocytes on the 14th day FA=(3·0+2·16+1·38+0·26)/80=0,87;

B-lymphocytes on the 14th day FA=(3·0+2·16+1·34+0·30)/80=0,83.

The function of the graft was stabilized, the marked increase in diuresis: 14, 16, 18 days - 600, 1100, and 2100 ml creatinine Levels of plasma 14, 16, 18, the day was 0.65; 0,49; 0.28 mmol/l and reached a rate to 25 days.

15.11.07 operation: removal of the peritoneal catheter. 22.11.07 removed the stent from urethra of neurotransplantation.

29.11.07 the patient was discharged with satisfactory function of the renal graft.

Example 2.

Patient M., 49, entered MONICA with a diagnosis of chronic glomerulonephritis, chronic renal failure-end stage, arterial hypertension, myocardial, the status of peritoneal dialysis. Ill for 25 years, for the treatment of peritoneal dialysis since February 2004

Transplantation of cadaveric kidneys made 13.07.2007. Donor male, 22 years old, cause of death was traumatic brain injury. Time primary heat ischemia 20 min Term conservation 30 hours.

Immunosuppressive therapy consisted of Cya, prednisolone, metypred, CellCept, zenapax. Function delayed graft, continued treatment with peritoneal dialysis. Diuresis on 1, 3, 5, 7, 9, 11 day amounted to 400, 500, 350, 300, 400, 400 ml, respectively. The level of plasma creatinine 1, 3, 5, 7, 9, 11 day 1,5; 0,97; 0,92; 0,88; 0,95; 0,88 mmol/l, respectively.

On day 7 after allotransplantation was performed phase morphometry T - and b-lymphocytes of peripheral blood. Sampling populations of each type of lymphocytes ranged from 65 to 100 cells. Was determined by the height of each phase of the lymphocyte and the value of this indicator cell was assigned a corresponding coefficient actively the minute from 0 to 3. Counts the total number of lymphocytes and the number of cells in each group and calculated the functional activity:

T-lymphocytes on day 7 FA=(3·14+2·27+1·16+0·8)/65=1,72;

B-lymphocytes on day 7 FA=(3·16+2·40+1·8+0·8)/72=1,89.

20.07.07 performed a biopsy of the kidney transplant.

Histological examination of kidney biopsy: in two renal biopsies contained 6-8 glomeruli, scleratinian no. The proximal part of glomeruli with slightly advanced lumen, epithelium them in a state of protein malnutrition. The interstitium is marked with a diffuse-focal lymph-plasma-cell infiltration (i2) with a tendency to penetrate into the wall and the lumen of the tubules, there are tricks partial destruction of the walls of the tubules (t3). Arterioles are not changed. In one partially cut artery marks the boundary distance mononuclear cells. Conclusion: Acute rejection 1B according to the Banff classification.

10 day started antichrista therapy of methylprednisolone in a total dose of 2500 mg

The study of functional activity of cells 11 and 14 days showed the effectiveness of the treatment.

T-lymphocytes:

on the 11th day FA=(3·0+2·8+1·28+0·44)/80=0,55;

14 day FA=(3·0+2·10+1·30+0·32)/72=0,69;

B-lymphocytes:

on the 11th day FA=(3·12+2·52+1·18+0·18)/100=1,58;

14 day FA=(3·0+2·16+1·34+0·30)/80=0,83.

The function of the graft was stabilized, the marked increase in diuresis: 14, 16, 18 day - 000, 1700 and 2000 ml creatinine Levels of plasma 14, 16, 18 day amounted to 0.68; 0,29; 0.21 mmol/l and reached norms by 20 days.

Thus, the efficiency protivogradovoy therapy can be quantified on the dynamics of the values of the functional activity of lymphocytes. Reduction of FA up to 1.3-1.4, which corresponds to the values in uncomplicated post-transplant period, testifies to the effectiveness of the treatment.

A method for diagnosing rejection, kidney transplant, including the determination of the phase height PH of living of peripheral blood lymphocytes using phase-interference microscopy, the total number of lymphocytes in the sample, characterized in that to determine the number of lymphocytes with phase height PH ≤ 1.5 µm, 1.5 µm ≤ PH ≤ 2 µm 2 µm ≤ PH ≤ 2.5 μm, PH ≥ 2.5 μm, while choosing the activity coefficients of lymphocytes for each limit specified phase heights lymphocytes equal to k3=3, k2=2, k1=1, k0=0, respectively, the data and determine the functional activity of lymphocytes in the sample FA by the formula:
FA=(k3n3+k2n2+k1n1+k0n0)/n, where
n is the number of lymphocytes in the sample,
n3the number of lymphocytes with PH ≤ 1.5 mm
n2the number of lymphocytes with 1.5 µm ≤ PH ≤ 2 µm,
n1- the number of cells with 2 µm ≤PH ≤ 2.5 μm,
n0the number of lymphocytes with PH ≥ 2.5 μm,
k3, k2, k1, k0- the ratios of lymphocyte activity, and when the value of the functional activity of lymphocytes within FA=1,8-2,0 diagnosed with kidney transplant rejection.



 

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EFFECT: higher accuracy of detection.

FIELD: medicine, diagnostics.

SUBSTANCE: the present innovation deals with blood sampling, separating plasma against erythrocytes, moreover, in plasma on should detect activity of antithrombin III, proteins C and S, XIIa-dependent fibrinolysis and concentration of plasminogen obtained results should be expressed as relative units followed by calculating integral parameter that characterizes the state of anticoagulant-fibrinolytic potential (IPAFP) by the following formula: IPAFP = [(C1 + C2)/(C3 + C4)] x 100, where C1 - the ratio of observed value of antithrombin III activity to the value of inferior border of the range of analogous parameter norm; C2 - the ratio of observed value for the activity of proteins C and S system to the value of inferior border of the range of this parameter norm; C3 - the ratio of the value of inferior border of plasminogen concentration under normal conditions to observed value of analyzed parameter; C4 - coefficient calculated with the help of regression equation: C4 = 0.9 + (0.01 x X), where X - terms of lysis of patient's euglobulin clot/min, and at IPAFP value of 101.4 U and higher one should state anticoagulant-fibrinolytic blood potential to be in norm, in interval of 64.8 - 101.3 -as insufficient, and at 64.7 and below - as critical. The present method simplifies the procedure of evaluating the state of endogenous anticoagulants and activity of XIIa-dependent fibrinolysis.

EFFECT: increased diagnostic value of obtained results.

3 ex, 1 tbl

FIELD: medicine, laboratory diagnosis.

SUBSTANCE: method involves determination of the patient blood content of globulin-alpha 1, globulin-beta, globulin-gamma and the total bilirubin content followed by calculation of diagnosis indices for the patient (Y1, Y2, Y3) by using the computer program "Statistica 1.5" and introducing values X1, X2, X3 and X4 in computer wherein X1 means globulin-alpha 1 value; X2 means globulin-beta value; X3 means globulin-gamma value; X4 means total bilirubin value. Obtained values of diagnosis indices for the individual patient (Y1, Y2, Y3) are compared with average values of diagnosis indices (Y1', Y2', Y3') for different urogenital infections followed by comparison by sign and value. By the maximal coincidence of diagnosis index values for the individual patient with average diagnosis index values urogenital disease is diagnosed and the following diagnosis index average values are used: for chlamydiosis: Y1' = -2; Y2' = -0.1; Y3' = -0.2; for mycoplasmosis: Y1' = 2; Y2' = 0.8; Y3' = -0.04; for ureaplasmosis: Y1' = 2; Y2' = -1; Y3' = 0.02; for health persons: Y1' = -2; Y2' = 0.1; Y3' = 0.2. Invention provides the development of a method for express-diagnosis of infection at initial stage and diagnosis of atypical forms that occur in these diseases, and differential diagnosis of chlamydiosis, mycoplasmosis and ureaplasmosis. Invention can be used for carrying out the differential diagnosis of chlamydiosis, mycoplasmosis and ureaplasmosis.

EFFECT: improved method for express-diagnosis.

2 tbl, 4 ex

FIELD: medicine.

SUBSTANCE: method involves determining absolute value of ratio between lymphocyte number and absolute value of monocyte number in peripheral blood at the end of combine radiation therapy. The ratio is divided by 4.05. The result value being greater than 1, no disease relapse occurrence is predicted during the first observation year. The value being less than 1, tumor growth progress is stated and carcinoma relapse is predicted at the first year after treatment.

EFFECT: enhanced accuracy in detecting pathological process progress before observing clinical manifestations.

1 tbl

FIELD: medicine.

SUBSTANCE: method involves determining infrared radiation absorption coefficient in blood plasma in bandwidth of 1543-1396 cm-1. The infrared radiation absorption coefficient is determined in %. The value being equal to 29.7±1.1%, catarrhal cholecystitis is diagnosed. The value being 26.4±1.4%, phlegmonous cholecystitis is diagnosed. The value being 21.2±1.8%, gangrenous cholecystitis is diagnosed. The value being equal to 18.6±0.5%, gangrenous perforated cholecystitis case is diagnosed. The value in norm is equal to 32.4±0.8%.

EFFECT: high accuracy and specificity of diagnosis.

FIELD: biomedicine.

SUBSTANCE: the present innovation deals with biomedical measuring technologies, in particular, to those to detect bactericide activity of blood serum according to the level of its inhibiting impact upon luminescence intensity of sulfur-sensitive luminescent bacteria (ΣimpO) against control - luminescence intensity the same sulfur-sensitive luminescent bacteria that had no contact with blood serum (ΣimpK), then one should calculate the value of bactericide activity of blood serum by the following formula:

As sulfur-sensitive luminescent bacteria one should apply either natural or recombinant microorganisms being characterized by direct proportionality between intensity of decreased spontaneous bioluminescence level and degree of bactericide effect. For example, it is possible to apply Escherichia coli strain with genes of Photobacterium leiognathi luminescent system. The suggested method enables to shorten the duration for detecting bactericide activity of blood serum and decrease its labor intensity.

EFFECT: higher efficiency of detection.

1 cl, 1 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: method involves determining blood insulin I and thyroxin T content and phagocytic leukocyte activity (PLA). Activity coefficient is calculated on the basis of formula KA=IxPLA/T. KA value being found greater than 2.8 units, considerable amelioration treatment effect is predicted. The value being from 1.4 to 2.8 units, amelioration is predicted. KA being less than 1.4 units, lower treatment efficiency is predicted.

EFFECT: high reliability of prognosis.

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