Immunohistochemical features of gastrointestinal stromal tumors and their role for differential diagnosis and prognosis

The aim. To clarify all most important immunohistochemical features of gastrointestinal stromal tumors with different histological patterns and analyze the role of expression of Ki-67, MMP-9, VEGF and p16ink4A as a predictive markers of tumor progression. Materials and methods. The study is based on analysis of 100 primary GISTs for description of their morphological features and 36 GISTs taken from this 100 for study of prognostic markers. Results. All spindle cell GISTs have shown diffuse expression of CD117 in tumor cells. The levels of CD117 expression varied from strong expression (3+) until mild expression (1+). Strong expression were seen in 75,8 % of spindle cell GISTs. Epithelioid GISTs demonstrated heterognous moderate or mild expression of CD117. All primary epithelioid GISTs from patients that had relapse of tumor in period from 1 till 3 years demonstrated focal mild expression of CD 117 in tumor cells. Expression of DOG-1 were seen in all 100 cases of GISTs, that were included in our study. The strong expression of DOG-1 (3+) were seen in all 45 GISTs that had low mitotic rate (≤5 mitoses per 50HPF) and not associated with their histological pattern. GISTs with high mitotic rate demonstrated heterogeneous expression of DOG-1 in tumors: moderate expression (2+) with patchy areas of strong expression (3+). Expression of CD56 was not found in spindle cell GISTs, but single tumor cells of epithelioid GISTs that had high mitotic rate demonstrated expression of this marker. The average expression of p16ink4A were higher in tumors that gave relapses compared with tumors without relapses (50,3 % versus 5,7 % respectively, U-test=16.5; p≤0,01).The average expression of MMP-9 also were significantly higher in GISTs that gave relapses: 63,2 % compared with 13,4 % in GISTs without relapse (U-test=16; p≤0 ,01).The strong VEGF expression was found in 66,7 % of GISTs that had relapses and only in 8,3 % of GISTs without relapses. 50 % of GISTs without relapses was negative for VEGF. Finally, the average expression of Ki-67 were 13,4 % in GISTs with relapses and 8,7 % in GISTs without them (U-test=16; p≤0,01). Conclusion. We highly recommend using DOG-1 for epithelioid GISTs. Additionally in epithelioid GISTs can be used CD56 that can give focal positive reaction in some tumour cells. The following minimal panel of markers for differential diagnosis of spindled GISTs from other mesenchymal tumors of gastrointestinal tract is proposed: CD117, DOG-1 and SMA, where the first too markers will demonstrated the moderate or strong diffuse expression and SMA can be occasionally positive in some tumor cells. p16ink4A, ki-67, VEGF and MMP-9 can be used as additional prognostic markers in GISTs.

Development of methods of identification and quantitative determination of active substances in semi-solid dosage forms with sapropel extracts

For use in veterinary medicine for the treatment of the wound process and the prevention of mastitis, the composition and technology of an emulsion-based cream, conventionally called “Saprocream”, has been developed. Clinical trial of the drug “Saprocream” proved its effectiveness and safety for use in the healing of microtraumas, erosions and cracks (wounds) of the udders of cows. The aim of the research were was the standardization of the cream under the conventional name “Saprocream” for use in veterinary medicine as a wound healing agent, the development of methods of identification and quantification and their validation. Materials and methods. Test samples of emulsion cream type o / v, containing 15 % aqueous extract of sapropel (VES), 10 % oil extract of sapropel (OES), 6 % emulsifier No. 1, 1 % cetylstearyl alcohol, preservatives – 0.01 % nisin, 0.1 % euxil K 100 and purified water up to 100 g were made by phase inversion. The methods of pharmacopoeial articles of the European Pharmacopoeia of the 10th edition, SPhU 1.0, as well as industry standards were used to develop methods for identification and quantification of active pharmaceutical ingredients (API) in the developed cream and its standardization. Results. The chromatographic profile of the acetone extract from the cream coincides with the chromatographic profile of the reference solution ((PRS) β-carotene), which indicates the presence of substances of carotenoid structure. The absorption spectrum of hexane extraction from the cream in the range from 500 nm to 750 nm has a maximum absorption at a wavelength of 670±2 nm, which coincides with the maximum absorption of hexane solution OES, indicating the presence of chlorophyll. A method for quantifying the total mass fraction of humic acids (HA) has been developed and it has been established that the total mass fraction of HA in cream samples with sapropel extracts is 0.828 %. The results of the validation of the method show that there is a linear relationship between the concentration of the total mass fraction of HA and the mass of the AES sample with a correlation coefficient of 0.9981 (³ 0.9981). The developed technique is precise, because the value of the relative confidence interval is less than the critical value for the convergence of the results: D %=1.34≤1.60 and the criterion of insignificance of systematic error d=0.51 is fulfilled. Conclusions. An emulsion-based cream under the conventional name “Saprocream” has been standardized, methods for identification and quantification of active substances have been developed and validation of the developed methods for use in veterinary medicine as a wound-healing and anti-inflammatory agent has been carried out.

Substantiation of the composition of a semi-solid dosage form with a probiotic component for use in dermatology

One of the important issues in the pharmaceutical development of a semisolid preparation for dermal use is the scientific and experimental justification for choosing the base-carrier of active substances. The aim of this study was to experimentally substantiate the choice of rational combination of excipients in the development of a semisolid dosage form with a probiotic component for use in dermatology. Materials and methods. Hydrophilic gelling agents were used as excipients in the study: Sepiplus 400, Aristoflex AVC, Carbopol 934, hydroxyethylcellulose HEC, sodium alginate. Physical and chemical, pharmaco-technological and microbiological methods were used to select the optimal basis. Colloidal stability was determined using a laboratory centrifuge at a speed of 6000 rpm for 5 min, an electronic thermometer and a water bath laboratory. Thermostability was determined in the thermostat at (40±2) °C for 24 hours. Investigation of the rheological properties of the samples was performed using a Rheolab QC (Anton Paar, Austria) rheoviscometer using a system of coaxial cylinders C-CC27/SS. The kinetics of water absorption of the samples were studied by dialysis through a semipermeable Cuprophan membrane, Type 150 pm at a temperature of (34±2) °C. The number of viable lactobacilli cells was determined by surface seeding on Petri dishes with dense MRS medium. Research results. The results of the study of the number of viable lactobacilli cells showed that the lowest number of viable cells was in sample based on HEC. Tests for thermostability and colloidal stability showed that for sample No. 5 based on sodium alginate, stratification was observed in the experiment, which indicates its instability. These samples were excluded. Samples on Sepiplus 400 and Aristoflex AVC on a set of rheological features have advantages both consumer and technological; and sample on t Aristoflex AVC has the best indicators of osmotic activity and the viability of cells in the dynamics for this study. Conclusions. As a result of physical and chemical (organoleptic, colloidal and thermostability), pharmaco-technological (osmotic and structural and mechanical properties) and microbiological studies (number of viable lactobacilli cells) studies it was found that further work should be carried out with a sample based on gelling agent Aristoflex AVC, which has the most optimal performance in this development for a semisolid dosage form with a probiotic component for use in dermatology

«Terrible triad» of the shoulder. Biomechanicalsemi-natural modeling andjustificationto rotator cuff restoration

The aim of this study: was determine the force of tension and deformation of axillary nerve in rupture rotator cuff and paresis of deltoid muscle of the shoulder joint. Material and methods: Semi-natural modelling based on the axial scans spiral computed tomography of the intact shoulder joint was performed to determine the degree of traction load on the axillary nerve with distal displacement shoulder head and tendon rupture which paresis of the deltoid muscle. Result: The values of deformations for axillary nerve being at the limit of tissue strength at distal displacement of humeral head of the model by 50 %, progressively increased with increasing distal displacement of humeral head to 100 % of its diameter, reaching values 1.7 times higher than the strength nervous tissue. Conclusion: The progressive changes occurring in the axillary nerve under the action of traction loads, and as a consequence of its ischemia, over time can lead not only to demyelination, but also to the defeat of the axons themselves atrophy of its fibers. In turn, deltoid muscle atrophy increases the traction load on the affected axillary nerve, which forms a vicious circle. The only possible option to "break" the vicious circle is restore the stabilizing structures damaged during the injury, among which one of the most important is the tendons of the rotator cuff of the shoulder. Surgical restoration of the integrity rotator cuff of the shoulder reduces the traction load acting on the axillary nerve, which in turn significantly improves the conditions for reinnervation of the deltoid muscle.

Study of hepatoprotective effect of bearberry leaves extract under insulin resistance in rats

The aim of our study was to evaluate the antidiabetic and hepatoprotective efficacy of dry extract from bearberry leaves enriched with arginine in dexamethasone induced IR. Materials and methods. IR was induced in rats by low dose intraperitoneally injections of dexamethasone. Dexamethasone-induced IR in rats was treated by bearberry leaves extract enriched with arginine. Thus, animals were randomized into several groups including intact animals and animals, which administered reference compounds and medications. The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamine transferase (GGT) were determined in blood serum and liver homogenate, in addition, in blood serum we measured lactate dehydrogenase (LDH) activity and lactate level and glycogen content liver tissue. Also, for the purpose of our experiment, in liver tissue were determined: thiobarbituric acid reactive substances (TBARS), diene conjugates (DC), and reduced glutathione (GSH) content; and superoxide dismutase (SOD), glutathione peroxidase (Gpx), and catalase (CAT) activities. All indices were determined using generally accepted unified methods or commercially available kits. Results. Long-term dexamethasone administration led to an increase in AST, ALT and GGT overall activity in the liver homogenate and serum; this could be the result of increased permeability of hepatocyte plasma membranes, as well as their enhanced synthesis in the liver. Studied extract ameliorate these indices of liver injury. Evaluation of indices that reflected oxidative stress and the antioxidant system status in liver confirmed oxidative stress development in IR rats` liver. Administration of arginine enriched bearberry leaves extract decrease TBARS and DC content in liver tissue, at the same time, improve SOD, Gpx, and CAT activities and increase GSH content. Conclusions. Bearberry leaves dry extract enriched with arginine inhibit oxidative stress development, improve membrane integrity, and normalize some indices of carbohydrate metabolism, particularly glycogen content in liver and lactate level in blood.

Clinical and immunological efficiency of different therapy regimens in patients with infectious mononucleosis caused by Epstein-Barr virus

The aim of the study was to evaluate the effectiveness of ribonucleic acid in the correction of immune disorders in patients with infectious mononucleosis (IM) caused by EBV. Materials and methods. We examined 110 patients with a mean age of 23.3±4.2 years with IM, among whom women accounted for 52.7 % (n=58). The material for the study was the serum of patients obtained during the disease course. The set of tests of patients with IM included clinical and biochemical methods, enzyme-linked immunosorbent assay, polymerase chain reaction method, immunogram. Statistical processing of the obtained data was performed with “Statsoft Statistica v. 10.0 for Windows” using the methods of variation and correlation statistics. Results. The obtained results analysis revealed changes in the system of cellular and humoral parts of the immune system and the diversity of the immune response in patients with IM. The progressive nature of changes in immune parameters indicated the formation of secondary cellular immune imbalance, activation of the humoral link, a change in the balance of immunoregulatory mediators towards Th2 cells. Significant changes in the cellular immune system were observed in the acute period and were characterized by the increase in the number of cells with killer activity, namely mature T-lymphocytes (CD3+), cytotoxic T-suppressor cells (CD8+), cells expressing the activation marker CD25+ (IL-2 receptor), and by the Th1/Th2 ratio increase. The appointment of combination therapy including ribonucleic acid was accompanied by immunomodulatory and antiviral effects, that was reflected in a more pronounced positive dynamics of immunological parameters, namely in strengthening the proliferative response, compared with the group of patients receiving only basic therapy. Conclusion. The expediency of the combination therapy application: the drug Nuclex (ribonucleic acid) (250 mg) 2 capsules 3 times a day for 14 days and valacyclovir (500 mg) at a dose of 1000 mg (2 tablets) 3 times a day for 14 days, is justified for the correction of immune disorders in patients with IM caused by EBV.

Endovascular interventions in patients with multilevel steno-occlusive lesions of the infrarenal aortic arteries

Peripheral artery diseases are a worldwide medical and social problem. Approximately 30 % of patients with critical limb ischemia will undergo amputations and 25 % will die after 1 year. These patients require reconstructive and angioplastic interventions to preserve the limb. The aim of the study is to compare the effectiveness of different endovascular surgical technologies in patients with multilevel steno-occlusive lesions of the infrarenal aortic arteries. Materials and methods. Among 243 endovascular interventions, 51 patients were presented with multilevel steno-occlusive lesions of the infrarenal aortic arteries as a result of atherosclerosis obliterans. 42 patients (82.4 %) had two-level lesions and 9 (17.6 %) had three-level lesions. Results. Among 42 patients with a two-level lesion, 29 (56.9 %) ones had femoral arterial segment affection in combination with tibial artery affection. 13 (25.5 %) patients suffered from the lesion of the iliac segment in combination with the affection of the femoral segment arteries. In 9 (17.6 %) patients the lesions of three or more levels were determined: an iliac segment in combination with femoral and popliteal arteries – in 4 patients, and in 2 patients there were lesions of the femoral, popliteal and tibial segments, in 3 patients there was a lesion of the iliac, femoral, popliteal and tibial-foot segments in different degrees of severity. 7 balloon angioplasties and 13 stenting procedures were performed in two and three-level lesions in which the iliac arterial segment was affected. Only balloon angioplasty was used to revascularize the infraingvinal arterial segments. In the early postoperative period, the complications included thrombosis appeared in 6 patients. It was possible to restore the blood flow and save the limb only in 3 persons. For the other 3 ones, the attempts to restore the patency of the arteries were unsuccessful and resulted in the amputation of the lower extremity. Mortality in the early postoperative period was 3.9 % (2 death due to myocardial infarction). Conclusions. The persons with multilevel steno-obstructive lesions of the infrarenal aortic arteries are the most difficult category of patients with CLI. The surgical method of choice for patients with multilevel steno-occlusive lesions is the endovascular angioplasty with or without stenting.

Predictive significance of the life history of children with gastroesophageal reflux for predicting the development of arrhythmias and conduction disorders

The aim: to improve the early diagnosis of cardiac arrhythmias and conduction disorders in children with gastroesophageal reflux, by studying the prognostic significance of the life history in this group of patients. Materials and methods. 56 children aged 8 to 18 years, mean age 14,93±2,62 years were examined. All examined children were divided into two alternative groups: Group I (main) was represented by 28 children with gastroesophageal reflux in combination with cardiac arrhythmias and conduction disorders, and Group II (control) - 28 children with only gastroesophageal reflux without cardiac arrhythmias and conduction disorders. Patients underwent clinical, anamnestic (with a detailed study of life history) and instrumental studies (electrocardiography, Holter daily ECG monitoring, esophagogastroduodenoscopy). Results. When analyzing the prognostic significance of the patient's life history, it was found that its very high level was observed for the number of foci of chronic infection (I=6.0) and the frequent incidence of a child up to year old of life (I=3.05). High prognostic value was registered relative to the number of diseases suffered by the child per year on average (I=2.35), and moderate predictor properties are characteristic of chickenpox (I=0.89) and the number of chronic diseases at present (I=0.71). The duration of breastfeeding showed low (I=0.30) prognostic significance. Suffered previously bacterial infections did not have predictor properties (I=0.12). In general, the prognostic significance of the patient's life history was high (I̅=2.20). Conclusions. It was found that in general the prognostic significance of the patient's life history was high (I̅=2.20) It is determined that in favour of the development of cardiac arrhythmias and conduction disorders evidence: the number of foci of chronic infection ≥1-2; lack of frequent morbidity in a child up to a year of life; the number of diseases suffered per year ≤5; the presence in past medical history of chickenpox; the number of chronic diseases in a child at present ≤2 and short-term (≤9 months) breastfeeding. It has been established what denies probability of development of the researched pathology: absence of the centers of a chronic infection; the presence frequent incidence of a child up to year old of life; the number of diseases suffered per year≥6, and chronic diseases at present ≥3; longer (≥9 months) breastfeeding and no history of chickenpox

Evaluation of vestibular function in children with purulent media otitis by computer stabilometry

The aim of the research – assessment of vestibular function in children with acute purulent otitis media by computer static stabilometry compared with typically developed children of the same age. Materials and methods: 22 children with purulent otitis media aged 4 to 7 years and 30 practically healthy peers were examined by static stabilometry on the device “MPFI stabilograph 1” (Kharkiv, Ukraine). Indicators of variation and shape of the pressure center distribution, spectral-correlation indicators of stabilograms, as well as integral indicators of stability (length, velocity and angle of postural oscillations) were taken into account and the equilibrium functions of the two sensory states (open and closed eyes) are calculated in the StabiliS software. Results: purulent otitis media causes posterior displacement of the absolute coordinates of the pressure center. In the state with closed eyes, children with purulent otitis differ from their healthy coevals by 14 of 24 stabilometric parameters, including stabilogram length, speed, angle and amplitude of oscillations, giving way to them in the stability of equilibrium. Children with purulent otitis media are more sensitive to the visual canal of postural control than their coevals. Involvement of visual afferents in postural control significantly improves balance maintenance in children with purulent otitis by reducing deviations of postural oscillations, which is reflected in the probable changes of 9 out of 24 stabilometric parameters compared to control. Conclusions: In preschool age, purulent otitis media leads to latent vestibular dysfunction, which is manifested by a violation of the stability of the vertical posture in a state with closed eyes and is compensated by visual-vestibulo-proprioceptive integration in the process of postural control.

The prospects for NT-pro BNP level determining in physical activity conditions in patients during the rehabilitation period after the myocardial infarction, complicated with decompensated heart failure

The aim of the study was to determine the futility of NT-pro BNP level analysis during dosed physical exercise for diagnosing decompensated HF in the postinfarction period and the possibility to determine the HF functional class. Materials and methods. 160 patients with previous MI were examined, patients’ average age was (56.67±5.72) years. The patients were randomized in 2 groups dependently on the presence of the signs of II А-B according to V. Kh. Vasylenko and M. D. Strazhesko classification and FC III-IV (according to NYHA) decompensated chronic HF. Group I consisted of 120 patients with Q-QS and non-Q MI; group II (the control one) had 40 patients with MI without signs of decompensated HF; 20 healthy people made up a norm group. Results. Signs of coronary insufficiency such as dizziness and signs of poor peripheral perfusion (cyanosis/paleness) occurred in 104 (86.7 %) and 79 (65.8 %) cases that differ from the same values in the second group 4 (10.0 %) and 14 (35.0 %) respectively. 88 (73.3 %) patients with the signs of decompensated HF felt general weakness, fatigue, and expressed a request to stop the test. The HR in the postinfarction patients with the signs of decompensated HF was before the exercise (88.2±3.18) bpm (p≤0.05); in the patients with past MI without the signs of decompensated HF, it was (75.32±3.41) bpm (p≤0.05); and in the practically healthy people, it was (77.73±3.02) bpm. We found out a reverse moderate correlation between the distances that had been covered by a patient during the 6MWT NT-proBNP level in blood serum of examined patients. Conclusions. Inadequate response to dosed physical exercises and increased immunological parameters as quantitative HF markers could be useful not only for diagnostics, but also for the risk stratification for decompensated HF that had occurred in patients on different rehabilitation stages after an acute MI.