Method for prediction of haematogenic metastasis of diffuse gastric cancer
SUBSTANCE: what is presented is a method for prediction of haematogenic metastasis of diffuse gastric cancer by immunohistochemistry of the primary tumour preparations. For the purpose of the study, the antibodies Cyclophilin A Antibody (CypA) clone NBP 1-54388 at the working dilution of 1:1200. If there is a positive CypA cytoplasmic expression in the primary tumour cells, a low risk of haematogenic metastasis is predicted.
EFFECT: invention enables predicting the low risk of haematogenic metastasis accompanying diffuse gastric cancer and avoiding excessive adjuvant therapy.
2 dwg, 2 tbl, 2 ex
The invention relates to medicine, specifically to Oncology, and relates to methods for predicting low risk of hematogenous metastasis in the diffuse type of gastric cancer.
Currently one of the most important histological classification of gastric cancer is the classification proposed by Lauren P. separation of tumors in the intestinal and diffuse types . Despite significant advances in diagnosis and prognosis of the disease in cancer of the stomach are still high morbidity and mortality from this disease .
The most significant prognostic parameters, predicting hematogenous the dissemination, which is one of the most frequent causes of death of patients with gastric cancer, consider the depth of germination the walls of the body, intravascular invasion, the severity of angiogenesis in the tumor site and lymphogenous metastasis[3, 4, 5].
Closest to the present invention is a method for determining indicators five-year survival rate and intensity of expression in the primary tumor indicator of apoptosis p53 proposed IkeguchiM., Cai J., N. Yamane et al. (1999). At high rates of expression of the five-year survival rate is 82%, with a low of 60%. The known method is not sufficiently accurate and informative in terms of forecasting emetogenic metastasis in diffuse gastric cancer the Disadvantage of this method is the fact, the study is carried out on the total group of patients with gastric cancer without specifying histotype tumor, does not specify a particular boundary values relating to "high" and "low" . In addition, five-year survival is a broad concept, including cases of occurrence of deaths not only from distant metastasis.
A new technical challenge - improving the accuracy and informative way.
To solve the problem in the method for predicting hematogenous metastasis in the diffuse type of gastric cancer by immunohistochemical studies of drugs of the primary tumor for research use antibodies Cyclophilin A Antibody (CypA)clone NBP1-54388 when working dilution of 1:1200 and in the presence of positive cytoplasmic expression of Sura in the cells of the primary tumor predicts a low risk of hematogenous metastasis.
The method is as follows. In patients found in the operational material diffuse type gastric cancer performed immunohistochemistry preparations of the primary tumor. For detection of expression of cyclophilin A use antibodies Cyclophilin A Antibody (CypA) clone NBP1-54388, working dilution 1:1200, firms Novus, USA. Detect the presence of cells with positive cytoplasmic expression of CypA in primary tumors with what Olson magnification (×400). In cases with positive expression of CypA predict a low risk of hematogenous metastasis.
The essence of the proposed method is illustrated by the following examples.
Example 1. Patient S., 43, with diffuse type gastric cancer without preoperative chemotherapy, stage T2N3M0. The study of the risk of hematogenous metastasis according to the proposed method. When immunohistochemical study was determined by the positive expression of CypA in the cells of the primary tumor. Within 60 months of follow-hematogenous metastasis has not occurred.
Example 2. Patient O., 31, with diffuse type gastric cancer without preoperative chemotherapy, stage T2N3M0. When immunohistochemical study was determined by negative expression of CypA. 24 months after surgery the patient appeared hematogenous metastases in the liver.
The selection marker driven by the study of literature data, patent sources and analyzed clinical observations. Cyclophilin A (CypA) is a protein localized in the cytosol, one of the representatives of the family of immunophilins, catalyzes the reaction of CIS-TRANS isomerization of X-Pro peptide bond . It is shown that this protein is involved in regulation of cell growth and differentiation . In addition to the cytosolic form, there is secretiruema form, which sintesio the t activated macrophages . The extracellular form of CypA acts as an agent of chemotaxis regulating the migration of monocytes, neutrophils, eosinophils and T-cells . This protein possesses immunomodulatory activity, stimulating the maturation and antigen presenting function of dendritic cells . In recent years there have been many studies that have revealed the overexpression of CypA in malignant tumors of various localizations [7, 8]. However, the exact role of CypA in carcinogenesis is not fully clear.
Criteria of the proposed method are selected on the basis of data analysis of clinical studies
The study included 29 patients with intestinal and 17 with diffuse type gastric cancer stage T1-3N0-3M048% men and 52% women. The average patient age of 59.5±10,1 years. Preoperative treatment of patients was not performed. The operation was carried out in volume Subtotal distal gastrectomy, gastroektomii proximal gastrectomy, Subtotal palliative gastrectomy surgery Billroth 2. The follow-up period was 5 years.
Histological type of gastric cancer was defined according to the classification proposed by Lauren P., 1965. Immunohistochemically the study was performed according to standard methods. For detection of expression of cyclophilin A (CypA) was used antibodies Cyclophilin A Antibody (clone NBP 1-543 88) (razocherovanie 1:1200) company Novus, USA. We determined the presence of cells with positive cytoplasmic expression of the studied marker at large magnification (h). Statistical processing of results was performed using the software package Statistica 6,0 for Windows.
In groups of patients with intestinal and diffuse types of gastric cancer was evaluated expression of CypA (figure 1, 2). The frequency of occurrence of cases with positive expression of CypA did not differ in patients with intestinal and diffuse types of gastric cancer (table 1).
In patients with diffuse type tumors hematogenous metastases were detected less frequently with positive expression of CypA, while in the intestinal type of gastric cancer, the incidence of distant metastases was not associated with the presence of CypA expression (table 2). The sensitivity of the method was 40%, the specificity determining the proportion of persons with a low risk of hematogenous metastasis - 100%.
Thus, the proposed method allows us to predict a low risk of hematogenous metastasis and to avoid excessive aggressive adjuvant therapy.
Sources of information used in the preparation of descriptions:
1. Lauren P. The two histological main types of gastric carcinoma: diffuse and so-called intestinal-type carcinoma [Text] / P. Lauren // Acta. Pathol. Environ. Scand. - 1965. No. 64. - P.31-49.
2. Tenderenda M. Expression of CD34 in gastric cancer and its correlation with histology, stage, proliferation activity, p53 expression and apoptotic index / M. Tenderenda, P. Rutkowski, D. Jesionek-Kupnika, R. Kubiak // Pathol Oncol Res. - 2001. - Vol.7(2). - P.129-34.
3. Fischer G. Determination of enzymatic catalysis for the cis-trans-isomerization of peptide binding in proline-containing peptides [Text] / G. Fischer, H. Bang, C. Mech // Biomed. Biochim. Acta. - 1984. - Vol 43. - P.1101-1111.
4. Bharadwaj, U. Effects of cyclophilin A on myeloblastic cell line KG-1 derived dendritic like cells (DLC) through p38 MAP kinase activation [Text] / U. Bharadwaj, R. Zhang, H. Yang, M. Li, L.X. Doan, C. Chen, Q. Yao // J. Surg. Res. - 2005. - Vol.127. - P.29-38.
5. Sherry C. Identification of cyclophilin A as A proinflammatory secretory product of lipopolysaccharide-activated macrophages [Text] / B. Sherry, N. Yarlett, A. Strupp, A. Cerami // Proc. Natl. Acad. Sci. - 1992; - Vol.89. - P.3511-3515.
6. IkeguchiM., Cai J., N. Yamane et al. // Cancer (Philad.). - 1999. - V.85. - P.2329-2335.
7. Lee J. Role of Cyclophilin A during Oncogenesis / J. Lee // Arch Pharm Res. - 2010. - VoL 33. P.181-187.
8. Obchoei S. Cyclophilin A enhances cell proliferation and tumor growth of liver fluke-associated cholangiocarcinoma / S. Obchoei, S. M. Weakley, S. Wongkham, Wongkham C, Sawanyawisuth K, Q. Yao, C. Chen // Mol Cancer. - 2011. - Vol.10. - P.102.
Figure 1. The expression of CypA in cribrosa structures in intestinal type gastric cancer (HC. ×400).
Figure 2. The expression of CypA in discrete tumor cells with diffuse type gastric cancer (HC. ×400).
Table 1. The frequency of occurrence of cases of CypA expression in tumor cells in intestinal and diffuse type of gastric cancer.
Table 2. The relationship of CypA expression in tumor cells with the presence of hematogenous metastases in the intestinal type of gastric cancer.
|The same type of cancer is the RCU||The expression of CypA (number of patients, ach, %)|
|Intestinal||3/29 (10%)||26/29 (90%)|
|Diffuse||2/17 (12%)||15/17 (88%)|
|Expression of cyclophilin A||Hematogenous metastases (number of patients, ach, %)|
|not (M0)||there are (M+)||not (M0)||there are (M+)|
|intestinal type||diffuse type|
|negative||3/3 (100%)||0/3 (0%)||0/2 (0%)||2/2 (100%)|
|positive||22/26 (85%)||4/26 (15%)||12/15 (80%)||3/15 (20%)|
|χ2=0.53; p=0,46||χ2=5,4; p=0.01|
A method for predicting hematogenous metastasis in the diffuse type of gastric cancer by immunohistochemical studies of drugs of the primary tumor, characterized in that for carrying out studies using antibodies Cyclophilin A Antibody (CypA) clone NBP 1-54388 when working dilution of 1:1200, and in the presence of positive cytoplasmic expression of CypA in the cells of the primary tumor predicts a low risk of hematogenous metastasis.
SUBSTANCE: cardiac recipient's blood plasma placental growth factor (P1GF) is measured prior to grafting and within 24 to 36 months after grafting. If observing the P1GF level increased more than by 60% of the pre-grafting values, the development of irreversible graft dysfunction is predicted.
EFFECT: it is possible to predict the remote development of irreversible graft dysfunction in the recipients after cardiac graft surgeries.
SUBSTANCE: single-domain antibody (nanoantibody) is proposed, which specifically binds a carcino-embrional antigen (CEA) of a human being and characterised by a full amino acid sequence. Also the method is considered to detect a CEA protein in biological fluids and tissues of a human being with usage of a nanoantibody according to the invention.
EFFECT: invention may find further application in diagnostics and therapy of cancer.
3 cl, 4 dwg, 4 ex
SUBSTANCE: single-domain antibody (nanoantibody) is proposed, which specificaly binds a mucin 1 protein (MUC1) of a human being and characterised by a full amino acid sequence. Also the method is considered to detect a MUC1 protein in biological fluids of a human being with usage of a nanoantibody according to the invention.
EFFECT: invention may find further application in diagnostics and therapy of cancer.
3 cl, 3 dwg, 4 ex
SUBSTANCE: invention refers to medicine, namely endocrinology, and concerns predicting the clinical effectiveness in diabetic polyneuropathy in the patients suffering type 2 diabetes and dyslipidemia. That is ensured by evaluating the intensity of diabetic polyneuropathy manifestations taking into account neuropathic dysfunctional score, evaluating triglycerides and glycolised blood hemoglobin before the treatment; the derived data are used to specify a therapeutic approach followed by further estimation of a probability of a successful correction of diabetic polyneuropathy by formula
EFFECT: method provides the improved clinical effectiveness ensured by the individual therapeutic approach based on the lipid and carbohydrate metabolism of a specific patient.
SUBSTANCE: diagnosticum comprises human lymphocytes to identify donor-specific major histocompatibility complex antigen antibodies during the post-transplant period, characterised by the fact that the diagnosticum comprises lymphocytes obtained from the multiorgan donor's spleen; a method for preparing a diagnosticum, predicting a rejection reactions following the transplantation from multiorgan donors, including a reaction to identify the donor-specific major histocompatibility complex antigen antibodies, characterised by the fact that the above diagnosticum is used. The diagnosticum contains T lymphocytes bearing the class I major histocompatibility complex antigens, and B lymphocytes bearing the class II major histocompatibility complex antigens in ratio suitable to provide an adequate antigen-antibody reaction in studying the donor-specific class I and II HLA antibodies in recipient's serum for a long transplantation period for humoral rejection diagnosis and evaluation of the immunosuppression therapy effectiveness.
EFFECT: preparing the high-quality diagnosticum.
4 cl, 2 ex, 2 tbl
SUBSTANCE: method is implemented by detecting a marker in herparinised venous blood by incubating the blood with a mycobacterial antigen material (a test sample) and with normal saline (a reference); thereafter the blood is examined to measure CD45+ lymphocytes of the fluoroisothiocyanate-labelled markers; the fluorescence of the labelled CD45+ lymphocytes in the test sample in relation to the reference enables to consider that the individual has been infected with the very species of mycobacteria with the antigen material has been used for the blood incubation in a test tube.
EFFECT: method enables supporting for the fact of the mycobacterial infection, identifying the infection agent species, assessing the vaccination effectiveness; the method can be implemented for a relatively short time.
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention discloses using the monoclonal antibodies (MCAs) produced by the hybridomas B/4H1 and 4H7 and directed to variable determinants as a part of influenza B virus proteins. The hybridoma B/4H1 and 4H7 strains are deposited in the Russian Collection of Vertebral Cell Cultures (RKKK P) under Nos. 719 D and 720 D, respectively. The two current influenza B viral evolution lines are identified with using the MCAs B/4H1 and MCAs 4H7 directed to variable determinants of Victorian-type and Yamagata-type influenza B hemagglutinin, respectively.
EFFECT: using the prepared MCA kit enables identifying the newly recovered influenza B viruses Besides, the MCAs under the invention may be used to study the antigenic structure and variability of this agent, to identify them as belonging to a specific evolution line; the MCAs may be also used to construct the diagnostic test systems.
3 cl, 12 dwg, 8 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention describes a chemically modified peptide that is a fragment of human protein S100β consisting of a sequence of 15 amino acids corresponding to the amino acids in position 18-32 of the amino acid sequence of protein S100β. The peptide is covalently bound to a spacer (Ahx) consisting of 2-aminoheptane acid or a similar compound, and covalently bound to the spacer by a biotin molecule (Bio). The peptide is described by general formula: A-Tyr1-Ser2-Gly3-Arg4-Glu5-Gly6-Asp7-Lys8-His9-Lys10-Leu11-Lys12-Lys13-Ser14-Glu15-E, wherein A represents Bio-Ahx, Bio-Acp, Bio or 0, and B represents 0 or -NH2. Bio means biotin; Ahx means a residue of 2-aminoheptane acid; Acp means a residue of 6-aminocapronic acid, 0 means the absence of any amino acid. A diagnostic technique and a method for prediction of melanoma provides measuring blood natural antibodies (nAB) to the peptide according to the invention with the stages of melanoma diagnosed by the antibody concentration as compared to the reference. The clinical outcome and therapeutic effectiveness in a craniocereberal injury are predicted by measuring the nAB to the peptide according to the invention. If observing the blood nAB level in the patients lower as compared to the standard reference, the favourable clinical outcome is diagnosed, and the therapy is stated to be effective.
EFFECT: invention may be used effectively as instant diagnosing of the disturbed blood-brain barrier and monitoring of the clinical effectiveness in CCIs and some cancers.
3 cl, 9 dwg, 1 tbl, 5 ex
SUBSTANCE: technique according to the invention involves the two-stage immobilisation of S and R brucellosis antigens on a tray by a non-specifically adsorbed mouse's monoclonal antibodies 2H2 and 2H8 in wells of the immunological tray (the first stage), specific binding S and R brucellosis antigens (the second stage) thereto, introduction and incubation of the analysed material (blood serum or milk) combined with introduction of mixed mouse's monoclonal antibodies 2H2 and 2H8 marked by horseradish peroxidase. The brucellosis agent antibodies are detected by being competitive with the antigen in binding on the well surfaces of the tray between the antibodies of the analysed sampled and fragment-marked monoclonal antibodies. A decreased level of the enzymatic signal in the analysed sample as compared with the negative reference testifies to contamination of the animal. Introducing the analysed sample into the two wells of the tray with S and R brucellosis antigens enables differentiation thereof for antibody specificity to any given form of the disease agent.
EFFECT: method involving the enzyme immunoassay enables increasing the effectiveness of health-improving measures, reducing the length of health-improvement in livestock farms with the negative brucellosis situation, dropping the disease incidence.
3 tbl, 3 ex
SUBSTANCE: ultrasound-assisted transcutaneous targeted needle liver-biopsy is used in the patients with acute destructive pancreatitis; that is followed by the immunohistological analysis of the biopsy micropreparations in parallel sections 5 mcm thick marked with monoclonal antibodies: CD 79a for identifying B-Lm and CD 68 for identifying MF. That is followed by optical microscopy at magnification x400 with morphometry of B-Lm periportal regions and MF lobe parenchyma; the results are further processed by automated image analysis systems. A relative area taken by the analysed cell elements in the micropreparation is determined by formula (A). If the value A for the B-Lm periportal regions falls within the range of 0.4 to 1.14, while the value A for the MF lobe parenchyma falls within the range of 0.89 to 2.95, a pre-infectious phase of acute destructive pancreatitis is stated. If the value A for the B-Lm periportal regions falls within the range of 1.41 to 4.98, while the value A for the MF lobe parenchyma falls within the range of 4.34 to 9.52, a phase of suppurative septic complications of acute destructive pancreatitis is stated.
EFFECT: using the method provides the well-timed correction of the therapeutic actions; the method is attended by a lower risk of the needle biopsy; it possess high accuracy and simplicity of implementation.
2 tbl, 2 ex, 6 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.
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.
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
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.
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
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.