Diagnostic Potential of Exosomal HypoxamiRs in the Context of Hypoxia–Sumoylation–HypoxamiRs in Early Onset Preeclampsia at the Preclinical Stage

As the search for non-invasive preclinical markers of preeclampsia (PE) expands, the number of studies on the diagnostic potential of exosomes is growing. Changes in the partial pressure of oxygen caused by impaired uteroplacental perfusion in PE are a powerful inducer of increased production and release of exosomes from cells, which also determine their cargo. At the same time, the expression pattern of oxygen-dependent microRNAs (miRNAs), called “hypoxamiRs”, is modulated, and their packing into exosomes is strictly regulated by sumoylation. In connection therewith, we emphasize the evaluation of exosomal hypoxamiR expression (miR-27b-3p, miR-92b-3p, miR-181a-5p, and miR-186-5p) using quantitative RT-PCR, as well as SUMO 1–4 and UBC9 (by Western blotting), in pregnant women with early-onset PE. The findings show that miR-27b-3p and miR-92b-3p expression was significantly changed at 11–14 and 24–26 weeks of gestation in the blood plasma of pregnant women with early-onset PE, which subsequently manifested. High sensitivity and specificity (AUC = 1) were demonstrated for these miRNAs in the first trimester, and significant correlations with a decrease in hemoglobin (r = 0.71, p = 0.002; r = −0.71, p = 0.002) were established. In mid-pregnancy, the miR-27b-3p expression was found to correlate with an increase in platelets (r = −0.95, p = 0.003), and miR-92b-3p was associated with a decrease in the prothrombin index (r = 0.95, p = 0.003). Specific exomotifs of studied miRNAs were also identified, to which the sumoylated ribonucleoprotein hnRNPA2/B1 binds, carrying out their packaging into exosomes. The expression of conjugated SUMO 1 (p = 0.05), SUMO 2/3/4 (p = 0.03), and UBC9 (p = 0.1) was increased in exosomes at early-onset PE, and the expression of free SUMO 1 (p = 0.03) and SUMO 2/3/4 (p = 0.01) was significantly increased in the placenta, as an adaptive response to hypoxia. Moreover, SUMO 2/3/4 was negatively correlated with miR-27b-3p expression in the placenta. In conclusion, the diagnostic potential of exosomal hypoxamiRs mediated by sumoylation may form the basis for the development of combined specific targets for the treatment of early-onset PE, as hnRNPA2/B1 is a target of miR-27b-3p, and its sumoylation creates miR-27b-3p–hnRNPA2/B1–SUMO 1–4 cross-talk.

Statistical characteristics of the components of coagulation hemostasis and the degree of oxygenation of rat blood in the normal and at different times of the experimental opioid effect

Background. The problem of non-drug use of opioid drugs occupies a significant place among the current problems of world medicine. Objective. Тo study the hematological parameters of coagulation hemostasis in the norm and the dynamics of their changes at different times of opioid exposure. Methods. The experimental study was performed on sexually mature, outbred male rats in the number of 80 animals, weighing 160-270 g, aged 4.5-7.5 months. Animals were injected intramuscularly with “Nalbuphine” once daily for one day (10-11 hours in the morning) for 98 days. The initial dose of nalbuphine during the first 2 weeks was 0.212 mg / kg, the next 2 (II - IV weeks) - 0.225 mg / kg, the next (IV - VI weeks) - 0.252 mg / kg, the next (VI - VIII weeks) ) - 0.260 mg / kg, the next (VIII - X weeks) - 0.283 mg / kg, the next (X - XII weeks) - 0.3 mg / kg, and during (XII - XIV weeks) - 0.454 mg / kg. Thus, the conditions for chronic opioid exposure were created. Animals are divided into 3 groups. The 1-st group of animals received Nalbuphine for 98 days, with subsequent collection of material (end of 2 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks and 14 weeks of experimental opioid exposure); The 2-d was the control group, which for 98 days received injections of saline intramuscularly in one period of time (10 - 11 o'clock in the morning). Blood sampling and study of hematological parameters of blood (platelet count, prothrombin time, prothrombin index, time of recalcification of stabilized blood, total fibrinogen, determination of hemoglobin, hematocrit) were performed according to conventional methods. Software R v 4.0.3 and RStudio v 1.2.5042 were used for statistical calculations and graphing. MSOffice Excel 2010 spreadsheets were used to generate the final tables and store the data. Results. The key to the dynamics of changes in the blood parameters of experimental animals was week 6 of the experiment, as most indicators had the highest dynamics up to 6 weeks including further indicators of stability, which was higher (fibrinogen and prothrombin index) or less (prothrombin time, recalcification time and hemoglobin) indicators of the control group. The blood hematocrit of the experimental animals decreased evenly at all study terms to a minimum value at 14 week, and the number of platelets evenly all times increased to a maximum value at the last term of the experiment. This trend in all indicators was confirmed statistically. Conclusion. Our research has made it possible to study first and then observe the dynamics of changes in coagulation hemostasis and the degree of oxygenation of blood in acute, subchronic and chronic periods of experimental opioid exposure with subsequent statistical comparison.

Features of Changes in Blood Parameters of Some Laboratory Syndromes and their Constellations in Patients with Liver Cirrhosis with Disorders of Bone Mineral Density

Introduction. Investigation of changes in certain laboratory blood parameters, and verification with their help of laboratory syndromes, and detection of constellations of laboratory syndromes in patients with liver cirrhosis (LC), which is possible for clinicians of all levels of medical care, need to clarify their features, which would suspect or verify disorders of bone mineral density (DBMD). The aim of the study. Investigate the features of changes in blood parameters of some laboratory syndromes and their constellations in patients with liver cirrhosis with disorders of bone mineral density. Materials and methods. 90 patients (27 women (30.0 %) and 63 men (70.0 %) aged 18 to 66 years) with LC were stratified into several groups: experimental (EG) (patients with LC with DBMD) (72 patients (80.0 %))), from which two subgroups were formed - EG A (patients with LC with osteopenia) (46 patients (63.9 %))), and EG B (patients with LC with osteoporosis) (26 patients (36.1 %)))) and the comparison group (CG) (patients with LC without DBMD) (18 patients (20.0 %))). Among the laboratory syndromes and their blood parameters were studied such as: cytolysis (increased in plasma alanine aminotransferase (ALT) and/or aspartate aminotransferase (AST)), mesenchymal-inflammatory syndrome (increased thymol test (TT) and/or gamma-globulins), hepatocellular insufficiency (decreased fibrinogen, prothrombin index (PTI), total protein, or albumin), cholestasis (increased alkaline phosphatase (AP), gamma-glutamyltranspeptidase (GGTP), total bilirubin), porto-systemic shunting (decreased sodium and/or potassium, and/or increased creatinine) and dyslipidemia (increased serum cholesterol, B-lipoproteins, triglycerides, low-density lipoprotein (LDL), decreased high-density lipoprotein (HDL)). The study was performed in three stages, the first of which studied the features of laboratory syndromes and blood parameters that characterize them, the second - constellations of laboratory syndromes, and the third - the simultaneous manifestation of a number of different laboratory syndromes in patients with LC with DBMD, osteopenia and osteoporosis. Each stage involved three steps: the first was to study the frequency of laboratory syndromes and their laboratory blood parameters in patients with LC and determine their share in each of the study groups, the second was to identify significant differences in the frequency of cases, and the third was to identify a direct stochastic relationship between the studied trait and DBMD, including osteopenia and osteoporosis. Results. After performing all three stages and each of the planned steps, it was found that laboratory syndromes and their constellations are more common among patients with bone lesions. However, there are statistically significant differences in the frequency of cases between EG and CG in the case of a decrease in HDL and the simultaneous manifestation of five different laboratory syndromes; between EG A and CG - decrease in HDL and simultaneous manifestation of two and three different laboratory syndromes; between EG B and CG - increase in AP, decrease in HDL and simultaneous manifestation of five different laboratory syndromes; between EG A and EG B - cytolysis syndrome, increase in AST, gamma-globulins, AP, constellation of cytolysis syndrome with hepatocellular insufficiency syndrome or cholestasis syndrome and constellation of all three syndromes. Confirmed direct stochastic association was found: with all manifestations of DBMD - increase in TT, a decrease in HDL, and constellations of cytolysis, mesenchymal-inflammatory and dyslipidemic syndrome, which may be supplemented by hepatocellular insufficiency syndrome and/or cholestasis syndrome; with osteopenia - increase in TT, increase in blood cholesterol, decrease in HDL, and constellations containing dyslipidemia syndrome and supplemented by mesenchymal-inflammatory, and/or cytolysis and/or hepatocellular insufficiency and/or cholestasis syndromes, and simultaneously only two laboratory syndromes in a patient with LC; with osteoporosis - increase in blood AST, TT, gamma-globulins, AP, decrease in PTI, potassium, HDL, the presence of cytolysis, cholestasis syndromes, constellations of cytolysis syndrome with hepatocellular insufficiency syndrome and/or cholestasis syndrome, which are supplemented by mesenchymal-inflammatory and dyslipidemic syndrome, and the simultaneous manifestation only three or five different laboratory syndromes. Conclusions. Laboratory syndromes, blood parameters that characterize them, and constellations of laboratory syndromes have certain features in patients with cirrhosis of the liver with disorders of bone mineral density, as in most cases are more common in patients with bone lesions and have a confirmed stochastic relationship with disorders of mineral density bone tissue in general, and osteopenia and osteoporosis separately. Keywords: cirrhosis, bone mineral density, osteopenia, osteoporosis, cytolysis, mesenchymal-inflammatory, hepatocellular insufficiency, cholestasis, porto-systemic shunting, dyslipidemia, alanine aminotransferase, aspartate aminotransferase, thymol test, total protein, albumin, gamma-globulin, fibrinogen, prothrombin index, alkaline phosphatase, gamma-glutamyltranspeptidase, bilirubin, sodium, potassium, creatinine, cholesterol, B-lipoproteins, triglycerides, low-density lipoproteins, high-density lipoproteins.

Esophageal Cancer: 10-Year Survival After Surgery

OBJECTIVE: 10-Year survival (10YS) after radical surgery for esophageal cancer (EC) patients (ECP) (T1-4N0-2M0) was analyzed. METHODS: We analyzed data of 551 consecutive ECP (age=56.5±8.9 years; tumor size=6±3.5 cm) radically operated (R0) and monitored in 1975-2021 (m=411, f=140; esophagogastrectomies (EG) Garlock=284, EG Lewis=267, combined EG with resection of pancreas, liver, diaphragm, aorta, VCS, colon transversum, lung, trachea, pericardium, splenectomy=154; adenocarcinoma=314, squamous=227, mix=10; T1=128, T2=115, T3=181, T4=127; N0=278, N1=70, N2=203; G1=157, G2=141, G3=253; early EC=109, invasive=442; only surgery=423, adjuvant chemoimmunoradiotherapy-AT=128: 5-FU+thymalin/taktivin+radiotherapy 45-50Gy). Multivariate Cox modeling, clustering, SEPATH, Monte Carlo, bootstrap and neural networks computing were used to determine any significant dependence.RESULTS: Overall life span (LS) was 1881.1±2230.6 days and cumulative 5-year survival (5YS) reached 52.1%, 10 years – 45.9%, 20 years – 33.7%. 184 ECP lived more than 5 years (LS=4308.7±2413.3 days), 99 ECP – more than 10 years (LS=5883±2296.6 days). 226 ECP died because of EC (LS=628.3±319.9 days). AT significantly improved 5YS (68.8% vs. 48.5%) (P=0.00025 by log-rank test). Cox modeling displayed that 10YS of ECP significantly depended on: phase transition (PT) N0—N12 in terms of synergetics, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), T, G, histology, age, AT, localization, blood cells, prothrombin index, hemorrhage time, residual nitrogen, protein (P=0.000-0.021). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 10YS and PT N0—N12 (rank=1), healthy cells/CC (2), PT early-invasive EC (3), thrombocytes/CC (4), erythrocytes/CC (5), lymphocytes/CC (6), eosinophils/CC (7), stick neutrophils/CC (8), segmented neutrophils/CC (9), monocytes/CC (10). leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).CONCLUSIONS: 10-Year survival after radical procedures significantly depended on: 1) PT “early-invasive cancer”; 2) PT N0--N12; 3) Cell Ratio Factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) EC characteristics; 9) tumor localization; 10) anthropometric data; 11) surgery type. Optimal diagnosis and treatment strategies for EC are: 1) screening and early detection of EC; 2) availability of experienced thoracoabdominal surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for ECP with unfavorable prognosis.

THROMBOELASTOGRAPHY IN ASSESSING THE HEMOSTATIC SYSTEM IN INTENSIVE CARE PATIENTS

The results of thromboelastography and standard coagulogram were analyzed in 35 patients aged from 18 to 86 who were treated in the resuscitation and intensive care unit. The majority of patients (34%) were hospitalized in the department with multisystem and concomitant injuries. The remaining patients were taken to the medical institution with different diagnoses (urolithiasis, liver cirrhosis, pancreatic lesion of various types, poisoning, peptic ulcer, sepsis). The data of coagulogram and thromboelastography at different stages of treatment were compared. In patients with the development of traumatic shock, the coagulogram parameters were changed to varying degrees depending on the stage of shock. At the first stage of shock, only an increase in soluble fibrin-monomer complexes by almost 2 times and a slight increase in fibrinogen dynamics were noted in the analysis. In a patient with stage 3 traumatic shock, the coagulogram parameters were within the normal range, but according to thromboelastography (EXTEM and FIBTEM tests), hypocoagulation due to the platelet link was noted. Only the coagulogram was evaluated in dynamics, hypocoagulation was noted in the indicators of internal and external hemostasis pathways: lengthening of the activated partial thromboplastin time, a decrease in the prothrombin index and an increase in the international normalized ratio, an increase in fibrinogen A and soluble fibrin-monomer complexes. In the group of male patients with closed craniocerebral trauma, an increase in soluble fibrin-monomer complexes in the coagulogram was always combined with changes in the FIBTEM test during thromboelastography. In most patients, no changes in the classical coagulogram tests immediately after the injury are noted. At this, thromboelastography makes it possible to make up for this deficiency at an earlier time, which indicates a high sensitivity of the method.

Changes in the blood coagulation system that determine postoperative complications in patients with non-tumor mechanical jaundice

The OBJECTIVE was to study the features of changes in the blood coagulation system that contribute to the development of postoperative complications in patients depending on the stage of non-tumor mechanical jaundice at admission.METHODS AND MATERIALS. A total of 537 patients with mechanical jaundice were examined and changes in the blood coagulation system were analyzed. Vascular-platelet hemostasis was characterized by the following tests: capillary resistance, the number of desquamated endothelial cells, the number of blood platelets. Plasma hemostasis was analyzed using activated partial thromboplastin time, plasma soluble fibrin level, thrombin time, prothrombin ratio, prothrombin index, and fibrinogen blood level. Then, XIIa-dependent fibrinolysis in the blood and the level of the fibrin D-dimer in the blood plasma were determined.RESULTS. It was found that in the first stage of mechanical jaundice, with cholestasis, there were no changes in blood coagulation system that go beyond the normal limits. In the second stage, during cytolysis of hepatocytes, hyperbilirubinemia and hypertransaminasemia contribute to the activation of platelet first, and then plasma hemostasis. In the third stage (cholangitis), the death of endotheliocytes increases and there is a deficiency of blood coagulation factors due to their consumption and increased fibrinolysis.CONCLUSION. In the stage of cholestasis in patients with non-tumors mechanical jaundice, the parameters of the coagulation system remain within the reference values. In the stage of cytolysis, as endotheliotoxicosis increases, platelet and plasma hemostasis begins to activate, which can lead to thrombosis and thromboembolism in vital organs. In the stage of cholangitis, further activation of plasma hemostasis causes hemorrhagic syndrome. The occurrence of the described disorders in blood coagulation system with the progression of MJ dictates the need to monitor the changes in the blood coagulation system and their correction for the prevention of intra-and postoperative complications.

Clinical and laboratory manifestations in critically ill patients with COVID-19 and deceased people

This article provides a review of the literature on the symptoms, laboratory blood values of critically ill patients who recovered and those who died of the new coronavirus disease COVID-19. Physicians should consider the following when predicting the course of the disease: in the first 3 days after admission, patients who recovered were slightly more likely to have leukocytosis and leukopenia, normal and increased lymphocyte counts; there were more individuals with increased number of band neutrophils, and patients who subsequently died were more likely to have normocytosis, granulocytosis, lymphopenia, thrombocytopenia, and higher erythrocyte sedimentation rate. The evaluation of laboratory indices in dynamics is of great importance for the prognosis: patients who recovered, on day 4–6 had a less pronounced growth of leukocytes and subsequently, on the contrary, their reduction; on day 4–6 of hospital stay, there was a decrease in the number of lymphocytes with subsequent growth; high creatine phosphokinase values at the beginning of hospitalization decreased significantly from day 7–9 to reference values; from the time of hospitalization, there was a decrease in lactate dehydrogenase content; the average prothrombin index tended to decrease, but within normal limits. Patients who died later, already from day 4–6 had an increase in leukocyte count, a decrease in lymphocyte level; thrombocytopenia was registered more often, which persisted with time and decreased significantly, especially after 9 days; in all periods of observation, erythrocyte sedimentation rate was higher (median of 30–40 mm/h); from day 7, there were significant fluctuations in maximum creatine phosphokinase values with their significant increase; at the beginning of hospitalization, these patients had higher lactate dehydrogenase levels compared to the first group and maintained their advantages during all periods of observation with significant fluctuations of maximal values; when comparing these patients by observation periods, there were slight fluctuations in the prothrombin index, which most often registered in about 80 % of patients with a subsequent increase after day 9, but within normal limits; also, at the beginning of hospitalization, there were significant fluctuations in the minimum prothrombin index towards very low rates.

CHANGES IN THE INDICATORS OF THE GENERAL BLOOD TEST AND COAGULOGRAM IN A MILD COURSE OF CORONAVIRUS INFECTION

In 2020 the pandemic of a new coronavirus infection spread in almost all countries of the world. The danger of this infection lies in the damage to the lungs, which can lead to fatal outcomes. The success of treatment is largely determined by the early diagnosis of the disease and its timely treatment. In order to find the ways of early laboratory diagnosis of coronavirus infection, we studied changes in peripheral blood indicators, coagulograms and C-reactive protein in 56 outpatient patients with mild course of coronavirus infection, whose average age was 49±2 years (among them, 23.2% were men and 66.8% were women). In all patients, coronavirus infection was confirmed by the detection of SARS-CoV-2 RNA by polymerase chain reaction (PCR) in nasopharyngeal or oropharyngeal smears. Signs of lung damage were detected in 66% of patients according to computed tomography findings. The number of red blood cells and the level of hemoglobin in all patients were normal. In 7% of patients, thrombocytopenia was detected, in 5.3% of patients – thrombocytosis. The most pronounced changes are foundin the leucoformula and indicators of the blood coagulation system. In 12.5% of patients without signs of viral pneumonia, leukocytosis was detected. All cases of leukopenia were noted only against the background of lung damage, at this, in 16% lymphopenia was found, 43% had relative lymphocytosis. The coagulogram of more than half of the patients with pneumonia showed shortening of blood clotting in the activated partial thromboplastin time (APTT) test, an increase in prothrombin index(PI) and the content of fibrinogen. The level of C-reactive protein was increased in only a quarter of patients with mild lung damage. Thus, changes in the leucoformula against the background of a tendency to hypercoagulation in blood clotting indicators should cause doctors' apprehensive attitude when treating a patient with clinical symptoms similar to COVID-19.

Ischemic stroke as a consequence of coagulopathy in coronavirus disease

Objective: to compare the state of the coagulation system in patients with ischemic stroke and coronavirus disease in those with ischemic stroke without coronavirus disease. Materials and methods. We examined 40 patients in the acute period of cerebral ischemic stroke, aged from 69 to 87 years (average age of 77.30 ± 2.08 years). They were divided into two groups: group I — SARS-CoV-2-positive, group II — SARS-CoV-2-negative. Group I consisted of 20 individuals (9 women and 11 men) aged 69 to 84 years (average age of 76.9 ± 3.0 years). Group II included 20 patients (10 women and 10 men) aged 72 to 87 years (average age of 78.5 ± 2.5 years). Results. Twelve (60 %) patients of group I had a moderate disease severity, 8 (40 %) — severe. The severity of neurologic deficits on the National Institutes of Health Stroke Scale did not differ significantly in patients of both clinical groups: in group I it was 14.0 ± 0.7 points, in group II — 10.00 ± ± 1.37 points, which corresponded to moderate stroke, the significance level p = 0.05. In the neurological status, motor and sensory deficits were observed in both groups in combination with speech disorders and ataxia. The average level of prothrombin index in group I was 101 ± 6 %, in II — 83 ± 2 %, p = 0.01. The level of fibrinogen in group I was 401.0 ± 18.6 mg/dl, in group II — 250 ± ± 12 mg/dl, which can lead to temporary hypercoagulation and the development of thromboembolism (p < 0.05). In group I, the level of D-dimer was 465 ± 8 ng/ml, in group II — 175 ± 4 ng/ml (p < 0.05). Conclusions. The data suggest that SARS-CoV-2-posi-tive patients with ischemic stroke compared to SARS-CoV-2-ne-gative ones with ischemic stroke are characterized by the presence of coagulopathy, as evidenced by significantly higher levels of fibrinogen, prothrombin index, D-dimer and moderate thrombocytopenia, as well as significantly higher levels of C-reactive protein that indicates the presence of an acute inflammatory process, which also causes thrombosis.

Peculiarity of clinical course of postpericardiotomy syndrome in different methods of surgery and postoperative antithrombotic therapy

Aim To compare features of the disease course and the effectiveness of nonsteroidal anti-inflammatory drug (NSAID) treatment of postpericardiotomy syndrome (PPS) in patients after coronary bypass (CB) surgery who were treated with antiplatelet drugs and in patients after surgical correction of heart valve disease (CHVD) who received the anticoagulant warfarin for prevention of thrombotic complications. Material and methods This study included 89 patients of whom 53 patients had underwent CB and 36 patients had underwent CHVD. At 15 [13; 15] days after surgery, the severity of inflammatory response, the state of coagulation hemostasis, and hematocrit were studied. At 5 days after the first test, blood count and measurement of C-reactive protein were repeated. Echocardiography was used to determine the presence and volume of pleural effusion. For prevention of thrombotic complications, antiplatelet drugs were administered after CB and warfarin was administered after CHVD. PPS was detected in 35 (66 %) patients after CB and 18 (50 %) patients after CHVD. The ibuprofen treatment (600 mg twice a day) was administered to all patients with PPS. If positive changes in inflammatory markers were absent during the NSAID treatment, ibuprofen was replaced with prednisolone 0.5 mg/kg body weight with subsequent laboratory and instrumental monitoring. Results Patients after CHVD treated with warfarin had higher values of international normalized ratio (INR) and activated partial thromboplastin time (aPPT) and lower values of prothrombin index (PTI), fibrinogen (p<0.001 for all), hemoglobin (p=0.0016), and hematocrit (p=0,0032) than patients after CB treated with antiplatelet drugs. 21 (40 %) patients with PPS required changing the anti-inflammatory therapy from ibuprofen to prednisolone. These patients displayed hypocoagulation, which was evident as reduced PTI (p=0.0023) and fibrinogen (p=0.0209), increased INR (p=0.0291) and aPPT (p=0.0416), and a higher incidence of pericardial effusion (p=0.0080). The insufficient effectivity of NSAIDs that required administration of prednisolone was more frequently observed in patients after CHVD (61 % vs. 29 %, р=0.037).Conclusion Hypocoagulation observed in patients after CHVD due to the anticoagulant treatment with warfarin was associated with more severe course of PPS and lower effectiveness of the NSAID treatment compared to patients after CB. This results in more frequent replacement of NSAIDs with glucocorticoids in the treatment of patients after CHVD.