Impact of Experimental Development of Arterial Hypertension and Dyslipidemia on Intravascular Activity of Rats' Platelets

Great interest is shown by researchers to functional and rheological features of basic regular blood elements. Platelets are among them and take the most active part in hemostasis at rather wide-spread nowadays cardio-vascular and metabolic diseases. The aim is to analyze dynamics’ strengthening of platelets' intravascular activity of rats in conditions of experimental consequent development of arterial hypertension and dyslipidemia. The study used 68 male-rats of Vistar line at the age of 2.5-3 months. Control group was composed of 33 animals. Experimental animals (35 rats) were developed at first – arterial hypertension (usage of cardioangiol pathogenic diet for 2 weeks and impact of cold at the end) and then - dyslipidemia (application of rich in calories diet at lowering of motor activity). The rats from experimental group were examined 5 times during the research. The rats from control group were examined twice - at the beginning and at the end of the experiment. While examining experimental and control rats we applied biochemical, hematological and statistical methods of investigation. At consequent arterial hypertension and dyslipidemia development experimental rats had gradual strengthening of lipids' peroxidation processes in plasma (acylhydroperoxides increased in 2.1 times) and platelets (acyl hydroperoxides increased in 1.4 times). Already at arterial hypertension development rats' blood was noted to have lowering of discocytes' quantity on 5.9%, deepening during the further dyslipidemia development on 7.4% more, and reaching 24.6±0.13%. This process was accompanied by gradual increase of activated platelets' sum on 75.7% during the whole period of investigation. The number of small, middle and large platelet aggregates, freely circulating in blood during modeling of double pathology, gradually increased in 2.8 times and in 10.3 times, respectively. Control rats had stable normal level of relevant biochemical and hematological characteristics. Subsequent development of at first arterial hypertension and then – dyslipidemia in rats gradually weakened their antioxidant protection of blood plasma and platelets and strengthened POL processes in them. It also strengthened lipids' peroxidation in them. Developing abnormalities gradually strengthened intravascular platelets' activity and their aggregative ability in experimental animals. The created model allowed tracking the earliest symptoms of strengthening of platelets’ intravascular activity against the background of AH and dyslipidemia development

INTRATUMORAL AMPLIFICATION HETEROGENEITY IN HER2/neu-POSITIVE BREAST CANCER MOLECULAR-GENETIC SUBTYPES

The defining feature of HER2/neu-positive Luminal B and HER2/neu-positive (non-luminal) subtype breast cancer is HER2/neu gene amplification and protein overexpression on cancer cell membrane. The HER2-targeted therapy is nowadays available for patients with HER2-positive breast cancer However, a significant fraction of HER2+ tumors acquire or possess intrinsic mechanisms of resistance, based on multiple factors, and genetic heterogeneity among them. The aim of our study was to quantify the heterogeneity of HER2/neu amplification in HER2/neu-positive Luminal B and HER2/neu-positive (non-luminal) subtypes of breast cancer. Material and methods. A retrospective analysis of 210 cases referred for dual probe fluorescence in situ hybridization (FISH) confirmation of an immunohistochemical equivocal 2+ result was performed. Results. Our results demonstrated a heterogeneous amplification pattern of HER2/neu gene, whose expression is a substantial cause of HER2/neu-positive Luminal B and HER2/neu-positive (non-luminal) subtypes of breast cancer, in 31 % of invasive breast cancer cases. As heterogeneous, we interpreted tumors containing cells with HER2/CEP17 ratio < 2 and gene copies 4 ≤ HER2/neu < 6, that is, those without HER2/neu amplification. The amount of heterogeneous tumors between HER2/neu-positive Luminal B and HER2/neu-positive (non-luminal) subtypes was not statistically significant. ROC analyses identified optimal cutoff point for HER2/CEP17 ratio as 2.6 for distinguishing heterogeneous tumors. Conclusion. The heterogeneity of HER2/neu amplification is determined by FISH in 31 % of cases and is independent of molecular breast cancer subtype. If a HER2/neu-positive breast cancer has HER2/CEP17 ratio ≤ 2,6, it contains minor subclones without HER2/neu amplification with a probability of 95 %. Our results demonstrated that HER2/neu amplification heterogeneity may be important for prognosis of survival and treatment decisions

Выявление ДНК вируса папилломы человека в поверхностной уротелиальной карциноме мочевого пузыря

An immunohistochemical research with antibodies to the mutant p53 protein and a revelation of the human papilloma virus DNA 16 and 18 types via in situ hybridization at the histological sections of the urothelial carcinoma are realized. The material of 44 patients with the superficial bladder cancer (Ta and T1 stages) and with a presence of the indirect signs of the viral infection was studied. 16 patients were included in the group of high risk recurrence, 13 patients were included in the group of the mean risk, 15 patients were included in the group of the low risk. HPV DNA was revealed in 12 of 44 cases only in patients of the mean and high risk groups (5 and 7 cases, respectively); all the positive results in the high risk group were with the HPV 16 type probe. Evaluation of the p53 protein showed a significant increase of its expression in the mean and high risk groups. p53 protein expression mean value was 23,67% in the low risk group, 36,53% in the mean risk group, 53,43% in the high risk group. Presence of HPV DNA was associated with the high p53 expression in the vast majority of cases.

Оценка экспрессии PD-L1 у пациентов с уротелиальным раком, имеющих противопоказания к назначению препаратов платины

Background. Urothelial cancer ranks 7th and 17th of all the malignant tumors in males and females, respectively. Development of new immunotherapeutic drugs provides new possibilities in treatment of such patients, especially the patient population in whom platinum‑based therapy is contraindicated. Administration of immunooncology drugs requires determination of PD-L1, for which various diagnostic systems are used. The question regarding correlation of results of determination of PD-L1 expression remains of concern.Materials and Methods. The study was performed on 100 samples of surgical and biopsy samples of urothelial cancer. Two clones of 22C3 and SP142 with corresponding detection systems were used for the study. PD-L1 expression was assessed in tumor and immune cells.Results. The study demonstrated a high correlation of negative PD-L1 tumor status determined using both diagnostic agents (92 % and 97 %) and low correlation of results of positive PD-L1 status (67 % and 43 %).Conclusions. Thus, if a negative result of PD-L1 status of urothelial cancer is obtained using any of the diagnostic agents studied, repeated test with the other antibody is not required. If positive status is obtained in one test, the patient may have a negative status in the other test, which allows recommending a repeated testing in borderline cases using a test, recommended for the medicinal product untended for treatment.Уротелиальный рак мочевого пузыря во всем мире находится на 7 и Введение. Уротелиальный рак во всем мире находится на 7 и 17 местах среди всех злокачественных опухолей у мужчин и женщин соответственно. Появление новых иммунотерапевтических препаратов открывает новые возможности для лечения таких пациентов, особенно учитывая группу пациентов, имеющих противопоказания к назначению препаратов платины. Применение иммуноонкологических препаратов требует определения экспрессии PD-L1, для которого используются различные диагностические системы. Актуальным является вопрос о сопоставлении результатов определения экспрессии PD-L1 разными методами.Материалы и методы. Исследования проведено на 100 образцах операционного и биопсийного материала уротелиального рака. Для исследования были использованы два клона антител 22C3 и SP142 с соответствующими системами детекции. Оценивалась экспрессия PD-L1 в опухолевых и иммунных клетках.Результаты. Исследование показало высокое совпадение негативного PD-L1 статуса опухоли, выявленного при использовании обоих диагностикумов (92% и 97%) и низкое совпадение результатов по позитивному PD-L1 статусу (67% и 43%).Выводы. Таким образом, при получении негативного результата PD-L1 статуса уротелиального рака любым из исследованных диагностикумов не требуется повторное тестирование с другим антителом. При обнаружении позитивного статуса по одному тесту пациент может иметь негативный статус по другому тесту, что позволяет рекомендовать повторное тестирование в пограничных случаях с тестом, рекомендованным для предполагаемого для лечения препарата

Сравнение иммуногистохимических тестов в рамках исследования CLOVER Российского общества клинической онкологии

Introduction. The goal of the CLOVER study performed by the Russian Society of Clinical Oncology, was a pairwise comparison of three validated PD-L1 immunohistochemical (IHC) tests (Ventana SP142, Ventana SP263, Dako 22C3) in the patient population with non-small cell lung cancer (NSCLC). This study is the first large Russian comparative study to evaluate PD-L1 expression levels using immunohistochemistry methods.Materials and methods. The study was conducted on 473 NSCLC samples from Biobank. The IHC tests were carried out with 3 antibody clones. Four trained pathologists independently evaluated the percentage of positively stained tumor cells (TC) and immune cells (IC). To assess the correlation of TC and IC between different runs and the prognostic values of one test for another, a concordant analysis was used.Results. The number of PD-L1‑positive cells (≥1 %) was higher among IC compared with TC in all three IHC tests. Pearson correlation coefficients (PCC) for TCs were 0.71, 0.87, and 0.75 for 22C3 / SP142, 22C3 / SP263 and SP263 / SP142, respectively. PCC values for ICs were 0.45, 0.61, and 0.68 for the same pairs. A high coincidence of positive and negative results (>91 %) was obtained between the staining with antibodies 22C3 and SP263 of immunooncological agents in the 1st line.Conclusions. The highest correlation between IHC tests was obtained by pairwise comparison of 22C3 and SP263. Clone 22C3 can be considered as a substitute for SP263 in the first-line treatment of NSCLC. Clone SP142 showed weaker expression in TC and IC compared to the other two tests in patients with non-small cell lung cancer.Введение. Целью исследования CLOVER, проведенного Российским обществом клинической онкологии, было попарное сравнение трех валидированныхиммуногистохимических (ИГХ) тестов PD-L1 (Ventana SP142, Ventana SP263, Dako 22C3) на одной и той же популяции пациентов с немелкоклеточным раком легкого (НМРЛ). Данное исследование — это первое крупное росийское сравнительное исследование по оценке определения уровней экспрессии PD-L1 методами иммуногистохимии.Материалы и методы. Работа проведена на 473 образцах НМРЛ, полученных из Биобанка. Иммуногистохимическое исследование проведено с использованием 3 клонов антител. Четыре подготовленных патологоанатома независимо оценивали процентное содержание положительно окрашенных опухолевых клеток (ОК) и иммунных клеток (ИК). Для оценки корреляции ОК и ИК между различными анализами и прогностических свойств одного теста для другого был проведен конкордантный анализ.Результаты. Число PD-L1-позитивных клеток (1% и более) было выше среди ИК по сравнению с ОК во всех трех иммуногистохимических исследованиях. Коэффициенты корреляции Пирсона (PCC) для ОК составили 0,71, 0,87 и 0,75 между 22C3/SP142, 22C3/SP263 и SP263/SP142 соответственно. Значения PCC для ИК составили 0,45, 0,61 и 0,68 для тех же пар. Было получено высокое совпадение положительных и отрицательных результатов (>91%) между окрашиванием, полученным с антителами 22C3 и SP263 для иммуноонкологических препаратов в 1 линии.Выводы. Наиболее высокая корреляция ИГХ анализов была получена при попарном сравнении 22C3 и SP263. Клон 22C3 можно рассматривать в качестве замены SP263 при лечении НМРЛ в первой линии. Клон SP142 показал более слабую экспрессию в ОК и ИК по сравнению с двумя другими анализами у пациентов с плоскоклеточным раком

ANTIPLATELET CONTROL OF VESSELS OVER THE MAIN BLOOD CELLS IN HYPERTENSIVES WITH DYSLIPIDEMIA IN COMPLEX THERAPY

The serious progress in dyslipidemia treatment (DL), incl. the cases of arterial hypertension (AH) comorbidity, has been statins introduction. It was found that the level of statins therapeutical effect grade significantly improves if there are hypolipidemic diet and tolerable exercises. However in this category of patients there was no assessment of the influence of the most active statins, including rosuvastatin (Rs) in combination with non-medication treatment, on the influence of antiaggregation properties of the vessels on the main blood cells.Aim. To assess in AH and DL patients, the opportunity of medical usage of the combination Rs with hypolipidemic diet and tolerable exercises, on antiplatelet properties of the vascular wall influence on erythrocytes, thrombocytes and neutrophils.Material and methods. Totally, 61 patients studied, with AH of 1-2 grades, risk 3, DL type IIb, middle age. Controls were 26 healthy persons of the same age. In the study, we used biochemical, hematological and statistical methods.Results. The enhancement of DL and AH patients aggregation of erythrocytes, thrombocytes and neutrophils, is mostly due to decreased antiplatelet control of vessel wall due to disorders of lipid metabolism, lipid peroxidation processes activation in plasma, decreased synthesis of nitric oxide and prostacyclin. As a result of the 6-month complex treatment, in AH with DL patients there is optimization of lipid content and of lipid peroxidation in plasma, with a 12-month achievement of complete normalization of antiplatelet abilities of vascular wall with its retention for at least 92 weeks of follow-up, if the therapy is continued.Conclusion. As a result of 1,5-month implementation of the complex treatment of AH and DL, there is optimization of lipid profile and peroxidation processes in plasma. Complete normalization of antiplatelet abilities of the vessel wall is reached by the 3rd moths with retention of the optimal values by the end of the study

Aggregation ability of the main blood cells in arterial hypertension and dyslipidemia patients on rosuvastatin and non-drug treatments

In Russia, there is still high prevalence of arterial hypertension (AH)comorbid with dyslipidaemia (DL), which increase aggregation properties of the blood cells. It seems rational to implement statins for rapid correction of aggregation properties of blood cells in AH with DL patients.Aim. To reveal the possibilities for complex influence of rosuvastatin and non-drug treatment on aggregation properties of blood cells in AH with DL patients.Material and methods. Totally, 61 patient with AH 1-2 grade studied, with risk 3 and DL IIb type, middle age. Controls were 26 clinically healthy volunteers of the same age. All patients were prescribed rosuvastatin and non-medication treatment. Antihypertension therapy — enalapril 10 mg b. i.d. Clinical and laboratory parameters registration was performed at baseline, in 6, 12, 52, 104 weeks after treatment start. For statistics, we used t- criteria by Student.Results. At the background of therapy, there signs of DL vanished, with more rapid increase of antioxidant plasma activity and with faster normalization of lipid peroxidation level. Rosuvastatin with non-drug treatment normalized aggregation properties of red blood cells, platelets, neutrophils in as low as 6 weeks of treatment by achievement of the optimum of their aggregation realization. The treatment established the effect for whole follow-up.Conclusion. To minimize the risk of hyperaggregation of blood cells and to relieve the lipid disbalance in plasma of AH with DL patients, it is recommended for them to take rosuvastatin for long time and with strict following the recommendations on non-drug interventions

Human papilloma virus DNA revelation in the superficial urothelial bladder carcinoma

An immunohistochemical research with antibodies to the mutant p53 protein and a revelation of the human papilloma virus DNA 16 and 18 types via in situ hybridization at the histological sections of the urothelial carcinoma are realized. The material of 44 patients with the superficial bladder cancer (Ta and T1 stages) and with a presence of the indirect signs of the viral infection was studied. 16 patients were included in the group of high risk recurrence, 13 patients were included in the group of the mean risk, 15 patients were included in the group of the low risk. HPV DNA was revealed in 12 of 44 cases only in patients of the mean and high risk groups (5 and 7 cases, respectively); all the positive results in the high risk group were with the HPV 16 type probe. Evaluation of the p53 protein showed a significant increase of its expression in the mean and high risk groups. p53 protein expression mean value was 23,67% in the low risk group, 36,53% in the mean risk group, 53,43% in the high risk group. Presence of HPV DNA was associated with the high p53 expression in the vast majority of cases

Infectious endocarditis in patients with injection drug use

In the article two clinical examples of infectious endocarditis (IE) in patients with diagnosis of “fever of unknown origin” admitted to the heart disease department from the surgical one are described