Method for predicting the relapse of serous ovarian cancer
FIELD: medicine, oncology.
SUBSTANCE: the present innovation deals with predicting the relapse of serous ovarian cancer after radical therapy before its clinical manifestation. The suggested innovation could be applied in daily practice of pathologoanatomic departments of hospitals, oncological dispensaries, specialized centers and research Institutes. The risk for the development of relapse is represented by a dichotomous variable calculated by the formula determined by logistic regression that includes the combination of several variables being morphological parameters of tumor, that is the area of tumor cell, average ploidy of tumor cells' nuclei, the level of p53 expression the level of expression of estrogen receptors, the level of PCNA expression. The innovation enables to predict repeated cancer growth according to morphological characteristics of primary tumor till clinical manifestation of the relapse.
EFFECT: higher accuracy and efficiency of prediction.
The invention relates to medicine, namely to Oncology, and is intended for the diagnosis before clinical manifestation of recurrent serous ovarian cancer after radical treatment.
Known clinical predictor of early recurrence and/or metastasis of cervical cancer , based on the determination of the ratio of the absolute number of lymphocytes to the absolute number of peripheral blood monocytes with concomitant radiation therapy, followed by dividing the ratio by 4,05. When the values of the result above one say no probability of relapse within the first year of observation, while values below one ascertain the presence of continued tumor growth and recurrence of cancer in the first year after treatment. However, this method can only be used when cervical cancer and is not applicable to carcinomas other localization, in addition, the result does not reflect the degree of probability of a positive or negative result at the end of the year relapse-free period.
Also known is a method for predicting the recurrence of breast cancer , based on biochemical studies of menopausal patients after combined treatment. Relapse ascertain if the reduction ratio of the concentration of astria the and to the concentration of estrone and estradiol in urine depending on the duration of recurrence-free period. It should be noted that the method is applicable only when breast cancer in postmenopausal women and uses the parameters defined for biochemical analysis.
The closest method described in . It lies in the fact that the forecast of development of recurrent endometrial cancer is set by the value of the ratio of cathepsin D and kislotostabilen inhibitors as determined by biochemical studies in tissue tumors, and endometrial. When the level of the coefficient is greater than the values typical for tissue intact endometrium, more than 2.4-2.8 times, predict the development of recurrence of endometrial cancer in the period up to 6 months.
The disadvantages of the method are the inability to use, ovarian cancer, prognostic judgment is not probabilistic in nature, and used options require specific biochemical methods.
The invention consists in that the forecast of the onset of relapse is established on the basis of the probability values calculated according to a formula determined using logistic regression. The formula is a combination of several variables, which is the morphological parameters of the tumor.
The technical result of the invention is the ability to forecast re-cancerous growth on morphologies the m characteristics of the primary tumor before the clinical onset of relapse, as well as the ability to use it when planning tactics of treatment of patients, as well as in the choice of treatment method.
The method is implemented as follows: define such parameters tumor size tumor cells, the average ploidy of the nuclei of tumor cells, the expression level of p53, the expression level of estrogen receptors, the expression level of PCNA.
The area of the cancer cell is measured by the eyepiece-micrometer or television image analyzer in μm2.
The average ploidy of the nuclei of tumor cells is measured by the photometer in units of the number of sets of chromosomes, C.
The magnitude of expression of p53 is expressed in points: the lack of p53 expression in tumor cells is scored 0; the expression of p53 detected in 1-10% of cells - 1 point; p53 is visualized in 11-50% of tumor cells - 2 points; p53 is defined in 51-100% of cancer cells is 3 points.
The level of expression of estrogen receptors was expressed in points: lack of expression of estrogen receptors in tumor cells is scored 0; the expression of estrogen receptors identified in 1-10% of cells - 1 point; estrogen receptors visualized in 11-50% of tumor cells - 2 points; estrogen receptors is determined in 51-100% of cancer cells is 3 points.
The magnitude of expression of PCNA (nuclear antigen of proliferating cells) is determined as a percentage of the number of PCNA - positive cells in the SDA is Oli.
Next, it calculates the probability of relapse according to the formula:
where X1 is the area of the cancer cell (μm2);
X2 - the average ploidy of the nuclei of tumor cells (C);
X3 - intensity labeling of p53 (in points);
X4 - intensity labeling of estrogen receptors (in points);
X5 - the value of the expression of PCNA (%).
While the final variable Y can take values in the range from 0 to 1. 0≤Y≤0,5 should be regarded as an indicator of low risk of resumption of tumor growth, while the 0.5≤Y≤1 testifies in favor of high risk of relapse.
The advantages of this method are the small number of morphological criteria, simplicity and low cost of the reagents to obtain the necessary parameters, the possibility of use in routine pathological practice and medical science. Examples of usage:
1. Patient M., 53 years of age, no history of the disease 4533, entered the Republican clinical Oncology dispensary mountains. Cheboksary September 25, 1998, in the direction of the precinct gynecologist. Upon receipt complained of heaviness in the lower abdomen, the increase in the abdomen. Heredity is not burdened, pregnancy was not. Objectively: the patient was in good condition, skin physiological painting,hemodynamic parameters within normal limits, vesicular breathing in lungs. Palpation of the abdomen is enlarged, soft, painless, during deep palpation by Obraztsov is determined by the free liquid in the abdominal cavity. At vaginal examination revealed a dense formation of granulomas in the pelvis on the left size of about 10 cm Pathological vaginal discharge no. Cytological examination of ascitic fluid in punctate Red space revealed atypical cells. Other changes in laboratory parameters are not set. At laparotomy diagnosed tumor of the left ovary solid character size 7 cm, capsule amazed. Produced hysterectomy with appendages and resection of the omentum with the following 6 courses of chemotherapy and hormonal therapy according to the standard scheme. Clinical diagnosis: Cancer of the left ovary, clinical stage Ic (FIGO), T1N0M0the ascites.
Histological examination was revealed serous histological type of carcinoma of the ovary with a low degree of histological differentiation. After conducting tsitomorfometricheskie study found that the average area of the tumor cells is equal to 82,35 μm. The average ploidy of the nuclei of tumor cells was 4,19 C. the expression of the antigen p53 was moderate (2 points), the expression level of PCNA - 70%, and the response to estrogen receptors weak (1 b the ll).
Thus, the calculated dichotomous variable Y:
Conclude that in this patient a very high risk of new cancer growth.
According to clinical observation of the patient obtained from the oncological dispensary, it is established that the patient M was diagnosed with a relapse of malignant neoplasms 9.04.2001 year.
2. Patient A., 58 years old, no history of 1598, came to the Republican clinical Oncology dispensary mountains. Cheboksary February 3, 1998, complaining of nagging abdominal pain, weakness. In anamnese 4 of pregnancy, heredity is not burdened. Objectively: the patient was in good condition, skin physiological painting, hemodynamic parameters in normal vesicular breathing in lungs. Palpation of the abdomen is enlarged, soft, painless, during deep palpation by Obraztsov is determined by the free liquid in the abdominal cavity. At vaginal examination in the area of the right epididymis diagnosed lumpy tumor site size up to 5 cm Pathological vaginal discharge no. When the puncture Red space in the peritoneal fluid revealed atypical cells. Other changes in laboratory parameters are not set. At laparotomy diagnosed tumor of the law is on solid ovarian character size 5 cm capsule amazed. Produced hysterectomy with appendages and resection of the omentum with the next 2 courses of chemotherapy and hormonal therapy according to the standard scheme. Clinical diagnosis: Cancer of the right ovary, clinical stage Ic (FIGO), T1N0M0the ascites.
Histological examination was revealed serous histological type of carcinoma of the ovary with a low degree of histological differentiation. After conducting tsitomorfometricheskie study found that the average area of the tumor cells is equal to 52,65 μm2. The average ploidy of the nuclei of tumor cells was 5,64 C. the expression of the antigen p53 - moderate (2 points), the expression level of PCNA - 100%, and the response to estrogen receptors high (3 points).
Thus, the calculated dichotomous variable Y:
Conclude that in this patient the risk of a relapse tumor growth is minimal.
According to clinical observation of the patient obtained from the Oncology center is established that the patient A. during the time of follow up relapse is not diagnosed.
1. The patent of Russian Federation №2247377 published 27.02.2005, Bulletin No. 6.
2. Application for invention No. 2003124101 published 27.01.2005, Bulletin No. 3.
3. The patent of Russian Federation №2250077, publish the consistent 20.04.2005, Bulletin No. 11.
A method for predicting the recurrence of serous ovarian cancer, which consists in the estimation of some parameters of the tumor and prognosis for him the possibility of recurrence of cancer, characterized in that it evaluated only morphological characteristics of the tumor and the likelihood of re-growth is determined by the formula
where X1 is the area of the cancer cell (μm), measured by the eyepiece-micrometer or television image analyzer, μm2;
X2 - the average ploidy of the nuclei of tumor cells (C) is measured by the photometer in units of the number of sets of chromosomes (C);
X3 - intensity labeling of p53 (point), the lack of p53 expression in tumor cells is scored 0; the expression of p53 detected in 1-10% of cells - 1 point; oncogene suppressor p53 is visualized in 11-50% of tumor cells - 2 points; p53 is defined in 51-100% of cancer cells - 3 points;
X4 - intensity labeling of estrogen receptors (point), no expression of progesterone receptor in tumor cells is scored 0; the expression of the progesterone receptor identified in 1-10% of cells 1 point; progesterone receptor visualized in 11-50% of tumor cells - 2 points; progesterone receptor is determined in 51-100% of cancer cells - 3 points;
X5 - the value of the expression of PCNA (nuclear ant the gene proliferating cells), %is determined by the number of PCNA - positive cells in the tumor, while the final variable Y can take values in the range from 0 to 1, 0≤Y≤0,5 should be regarded as an indicator of low risk of resumption of tumor growth, while the 0.5<Y≤1 testifies in favor of the high likelihood of recurrence.
FIELD: veterinary science.
SUBSTANCE: the suggested method deals with isolating lymphocytes out of peripheral blood and lymphoid organs to achieve their concentration up to working concentration followed by reaction of blast-transformation with mitogen as phytohemagglutinin, incubation of lymphocytes and registration of reaction results by radiometric method: concentration of lymphocytes should achieve working concentration value of 10-15x106 cells/ml, incubation of lymphocytes should be performed for 48-54 h at 37.5-40.0 C, index of lymphocytic stimulation should be calculated by the following formula:index of stimulation = (average amount of impulses/min in experiment) / average amount of impulses/min in control) The method provides increased efficiency in evaluating immune state in minks due to increased sensitivity in reaction of blast-transformation.
EFFECT: higher accuracy and efficiency of detection.
1 ex, 5 tbl
FIELD: biotechnology, in particular for biochemical, immunological and biological investigation and for production of insulin pharmaceuticals.
SUBSTANCE: semisynthetic human insulin is prepared by transpeptidation of swine insulin in presence of trypsin in mass ratio trypsin/swine insulin of 1:300-1000 and molar excess threonyn di-tert-butyl ester in relation to swine insulin, in aqueous-organic medium. Reaction is inhibited by pH reducing and reaction mixture dilution with water by 2-3 times. Obtained human insulin ester is purified by HPLC followed by providing of human crude insulin crystals and repeated HPLC-purifying. Method of present invention makes it possible to increase human insulin ester yield up to 98 % and decrease by-product contamination in finished product up to 0.9 %.
EFFECT: method of improved yield; product of improved purity.
FIELD: biochemical engineering.
SUBSTANCE: set is usable for testing antibodies in tumors, like radioactively labeled anti-CD20-conjugates. The conjugates are used for treating and diagnosing B-cell lymphoma tumors. All reagents are sterile.
EFFECT: wide range of functional applications.
103 cl, 38 dwg, 46 tbl
SUBSTANCE: method involves studying luminal-dependent iron-induced model system chemiluminescence on citrate-phosphate buffer, determining luminous sum and carrying out repeated model system chemiluminescence registration with chronic cholecystitis patient bile in remission state being added. The luminous sum being grown by 111-127% when compared to model system, stone formation in the gallbladder is to be predicted.
EFFECT: high accuracy of prognosis.
FIELD: medicine, medicinal microbiology.
SUBSTANCE: for detection of trichomonas forms method involves blood seeding in Teras nutrient medium with addition of human or equine serum blood inactivated at temperature +56°C for 5-10 min and antibiotics in the dose 500 U/ml. Culture is incubated for 2 days at temperature +37°C followed by reseeding in similar medium but containing human or equine serum blood inactivated for 15-20 min and antibiotics in the dose 250 U/ml and incubated again at temperature +37°C for 2 days. Then reseeding is carried out in medium containing serum inactivated for 30 min and without addition of antibiotics and samples are incubated at temperature +37° for 2 days. Also, 1-2 additional reseeding are carried out in medium with completely inactivated human or equine serum blood and without addition of antibiotics. Based on active budding or shizogony the conclusion is done about the presence of live atypical forms of trichomonas and in case the absence of multiplication - about the presence of degenerative destructing forms. Using the method allows detecting atypical forms of trichomonas.
EFFECT: improved detecting method.
1 tbl, 2 ex
FIELD: medicine, pharmacy.
SUBSTANCE: invention relates to study of antibacterial effect of ozonized physiological solution (OPS) and can be used in suppurative and combustion surgery. Method involves isolation of bacterial strain from patient blood with severe thermal trauma complicated by hospital infection and its parent properties are assayed. The strain is cultured on slant agar, poured with OPS in the ozone concentration 1, 3 and 5 mg/l and incubated in thermostat for 24 h. Standard suspensions containing 5 x 105 CFU/ml are prepared from diurnal cultures that are incubated in nutrient medium under condition of microbiological analyzer for 18 h. Then dynamic determination of optical density of medium is carried out in combination with plotting curves reflecting the growth of microorganisms and used them for evaluation of antibacterial effect of OPS on the microorganism strain. Also, elongation of LAG-phase, double time of microorganism counts and amount of microorganisms in 1 ml of nutrient medium are determined. Using the proposed method provides enhancing precision and information content of antibacterial effect of the ozonized physiological solution.
EFFECT: improved evaluation method.
1 dwg, 1 ex
SUBSTANCE: method involves producing sample A from patient blood for preparing leukocytic suspension, blood platelets suspension and medium containing no blood platelets. Physiologic saline and the sample A are placed into acoustic cells of two devices connected to computer for controlling biological fluids under temperature supported in them at 15-40°C. The temperature is lower in the first device. Ultrasound is let pass through them at frequency varying with in bandwidth 2-40 MHz and passage speed and transmission coefficient are concurrently measured in the sample A and then ultrasound passage speed is measured in the sample A during time interval lasting so long that leukocytic suspension, blood platelets suspension and medium containing no blood platelets subsequently substitute the physiologic saline in the first device and ultrasound propagation speed is measured in them. Hemoglobin concentration, erythrocyte, leukocyte and blood platelets number, hematokrit and erythrocyte sedimentation rate in whole blood are determined from the data received.
EFFECT: high accuracy of measurements.
FIELD: veterinary medicine.
SUBSTANCE: the method deals with preparing the mixture of microbial culture E.Coli, introducing it into experimental and control tubes, adding blood serum tested into experimental tubes, adding 24-h E.Coli culture into all tubes followed by mixing and measuring optic density of tubes' content at wave length being equal to 460 mcm followed by incubation for 3 h at 37° C to conduct additional measuring optical density and establishing bactericidal activity of blood serum by the formula. The innovation applies pre-inactivated (at 56° C) rabbit's blood serum obtained due to heating rabbit's blood in water bath for 2 h at 38° C and centrifuged for 15 min at 2500-3500 rot./min.
EFFECT: higher accuracy of detection.
FIELD: medicine, laboratory diagnostics, clinical gastroenterology.
SUBSTANCE: it is necessary to carry out morphological studying for 3 slices out of 3 mucosal departments of large intestine, not less to detect the presence of the focus of low-differentiated fibroblasts and xanthomatous or fatty cells in mucosal own plate. In case of their availability one should predict unfavorable flow of inspecific ulcerous colitis.
EFFECT: higher accuracy of prediction.
FIELD: medicine, laboratory diagnostics.
SUBSTANCE: the present innovation deals with detecting immunological disorders for matching pathogenetic therapy. In lesion foci, at the border with apparently healthy parts and in the nearest healthy parts of skin it is necessary to detect the value of microcirculation due to laser Doppler flowmetry and at maximal value of microcirculation in lesion focus one should detect affected processes of T-lymphocytes differentiation, and at minimal value of microcirculation in lesion focus one should state upon affected function of T-suppressors. Thus, the innovation enables to develop simplified and high-informative method for detecting the character of immunological lesions in children in case of atopic dermatitis.
EFFECT: higher efficiency.
SUBSTANCE: for the purpose to detect helminths and their larvae it is necessary to obtain microscopic preparations by mixing blood samples with anticoagulant K3-EDTA and physiological solution at 1:1 ratio, preparing the second solution based upon the first solution with 0.5%-NaOH and taking, correspondingly, at the ratio of 2:1 to incubate the solution obtained at 60° C for about 5-10 min, centrifuge it at 3000 rot./min for 5 min, remove supernatant, incubate the residue at 37° C for 10 min, treat the residue with 1%-Lugol's solution at the ratio of 1:1, prepare smears, dry them at 37 C for about 15-20 min and carry out microscopy at objectives 10 and 40. The present innovation enables to detect parasites in 100% of cases.
EFFECT: higher efficiency.
2 dwg, 2 ex
SUBSTANCE: preparations for carrying out microscopy should be prepared out of glucose-enriched supernatant, on mixing blood sample with anticoagulant K3-EDTA it is necessary to add glucose concentrated solution at not more than 10% against the volume of blood sample to create in blood sample glucose concentration of about 50-100 mM/l, incubate blood samples in a thermostat at 37° C for 15 min, take about 10-15 mcl out of the upper layer of supernatant to be applied onto a slide and covered with a cover glass to carry out microscopy at objectives being 10 and 40. The innovation enables to detect parasites in 100% of cases.
EFFECT: higher efficiency.
3 dwg, 2 ex
FIELD: medicine, otorhinolaryngology.
SUBSTANCE: the present innovation deals with treating diseases of the upper respiratory tract, at predicting pathological processes or diseases of nasal mucosa and paranasal sinuses and, also, with developing new methods for studying biological liquids. The method deals with sampling nasal secreta with a brush due to several light movements in area of nasal septum and inferior concha in anterior-posterior direction leaving about 1-1.5 cm against anterior edge, in the site of ciliated epithelium location followed by mixing it with 0.5 ml sodium chloride physiological solution, centrifuging for 15 min at 900 g, application of supernatant at the volume of 2.0-2.5 mcl onto glass surface as a drop and drying it due to wedge or terminal dehydration till obtaining the structure of solid phase. Thus, the innovation enables to obtain nasal secreta useful for morphological investigation due to above-mentioned types of dehydration.
EFFECT: higher efficiency.
2 cl, 2 dwg, 2 ex
FIELD: medicine, orthopedics, traumatology.
SUBSTANCE: one should measure the height of longitudinal pedal arch with the help of a foot-meter. In 2-3 min it is necessary to fulfill repeated measurement under conditions of maximal muscular relaxation. At decreased height of longitudinal pedal arch one should diagnose the availability of longitudinal platypodia.
EFFECT: higher accuracy of clinical diagnostics.