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The difference between the open-circuit redox potentials of a platinum electrode measured in blood plasma before and after the cryolysis of platelets depends linearly on the content of granule-rich platelets in the plasma. This phenomenon can be used to determine the viability of platelets.

We studied in vitro attachment, proliferation and survival of сadaver skin fibroblasts in collagen bands, dermal matrix and cancellous demineralized bone, enriched with platelet components. It was shown, that PRP enhanced revitalization of collagen grafts, especially followed by components of activated or freezed platelets. Fibroblasts had the best rate of proliferative activity in response to 150 pg platelet derived growth factor (PDGF), released from 100 million platelets with granules and measured for 100 × 103 cultivated cells. The use of platelet-derived material could increase fibroblast proliferation activity for 1.5-3 times in all types of collagen transplants without damage or decay of cell’s biological value.

The article presents a comparative analysis of the efficiency of allogeneic progenitor cells with different biological carriers for preparation of extensive traumatic wounds to autodermoplasty. As carriers were selected dermal matrix and collagen first type of human. The article presents the results of treatment of 43 patients. Based on these data the effectiveness of the combination of allogeneic progenitor cells and dermal matrix is demonstrated. This combination reduces the incidence of purulent complications and reduce the time of prepare wounds for autodermoplasty.

Morphofunctional evaluation of the effect of biological dressing with collagen-1 on healing of 3A degree burn wound in outbred and mutant Hr(hr)/Hr(hr)(hairless) mice was carried out by the histological method and optic radioautography. A pronounced stimulatory effect of the dressing on skin regeneration in mice was demonstrated. According to radioautography data, early dressing of the burn wounds in Hr(hr)/Hr(hr)mice led to active proliferation of epithelial cells in dermal cyst and vascular endotheliocytes. The possible mechanisms of the stimulatory effect of collagen-based dressing on wound healing are discussed.

A model of osteochondral intra-articular defect in rats is presented. During spontaneous healing, the stage of formation of granulation tissue is followed by its replacement with bone and fibrous tissue. Chondroinductive properties of collagen 1 sponge used for defect filling manifested in the formation of fibrous cartilage with fields of hyaline cartilage. Filling of the defect with collagen 1 sponge containing bone powder stimulated regeneration of the bone tissue and hyaline cartilage.

To evaluate patients' hemostasis after cardiac surgery using thromboelastometric and impedance aggregometry.66 patients were examined intraoperatively. Comparison group included 45 blood donors. Hemostasis was tested for thromboelastometricRotem Gamma with the assessment of external (exTem) and internal (inTem) pathways of coagulation tests performed detection of heparin (hepTem) and cytochalasin-D-inactivation of platelets (fibTem) to assess the level of fibrinogen. Collagen-induced platelet aggregation was determined in an aggregometer CHRONO-LOG (USA).Significant deviations of the parameters of hemostasis were detected in 52 of the 66 studied patients. In group-1 (23 patients) revealed a residual effect of heparin. The effect manifested prolongation CT (clotting time) inTem to an average of 241 +/- 15 s, compared with CT hepTem--181 +/- 7. Patients in this group were in need of additional administration of protamine sulfate. Postoperative bleeding and resternotomia were observed in 3 patients of group-1. In group-2 (25 patients) CT inTem was 216 +/- 21 with significantly fewer CT hepTem (272 +/- 26). The data indicated excess of protamine sulfate. Platelets aggregation decreased compared to the norm. According to the obtained results, the addition of protamine sulfate is not required, however, in 7 cases the protamine sulfate was administered in a dose of 8.9 +/- 0.8 mg in 6 cases resternotomiya required. In the third group (n = 6) bleeding was observed in 4 patients. The difference in CT-hepCT was significant. Significant variations were revealed in the tests of the activity of the extrinsic pathway of coagulation and cytochalasin-D-induced inactivation of platelets: exMCF- 42 +/- 2 mm (normal 57 +/- 15 mm), fibMCF 5.0 +/- 0.3 mm (norm 12.8 +/- 4.3 mm). The concentration of platelets and their aggregation activity was sharply reduced. Disorders of hemostasis in the third group, designated as dilution coagulopathy.Turning thromboelastometric and impedance aggregometry in the study of the coagulation profile of patients undergoing cardiac surgery in postperfusion period brings valuable information and allows a differentiated treatment of hemostasis disorders.

New effective method of active immunoprophylaxis of Bacillus Pynocyaneous in victims with open long bones fractures was elaborated. Anatoxin of Bacillus Pynocyaneous (15-30 mkg of protein/day) was injected into medullar canal of injured limb for 3-5 days after trauma. Injection of anatoxin was performed through microirrigator that was placed at the time of first osteosynthesis operation. Application of that procedure activated all components of immunity at early stages (by 7 day after injury) resulting in subsequent increase of immune response. Advantages of the given method in comparison with standard scheme of active immunoprophylaxis were confirmed by immunologic study data as well as by favorable clinical outcome of wound process.

The morphologic evaluation of adhesive activity of human thrombocytes using both vital dye of cells and registration of adhesion process with fluorescent microscope is proposed. The regression dependence between adhesive activity of human thrombocytes and morpho-functional and aggregative activity of thrombocytes is established. The blood samples of 100 blood donors, 100 patients with hematologic pathology, 110 patients of traumatology, 50 patients with acute exogenous intoxications and 20 patients with chronic trophic ulcers were analyzed. The drastic decrease of adhesive activity of human thrombocytes and number of adhesively active thrombocytes was established in circulating blood of patients with bleedings of various etiology and hematologic patients. The significant increase of adhesive activity of human thrombocytes and number of adhesively active thrombocytes in patients with thrombosis is established. The morphologic evaluation of quality of thrombocytes is effective in case of analysis of biologic value of all population of circulating cells.

Serum concentration of platelet-derived growth factor was measured in patients with trophic ulcers and proliferative activity of human fibroblasts was measured in wells with different content of added serum platelet-derived growth factor. In parallel, morphofunctional analysis of platelets from the whole blood of these patients was performed. A close correlation was found between morphofunctional parameters of blood platelets and serum concentration of plateletderived growth factor. Platelet-derived growth factor in concentrations below 150 pg per well stimulated fibroblast proliferation and preserved their viability. Platelet-derived growth factor in concentrations >200 pg per well suppressed fibroblast proliferation and impaired their viability.

We carried out a comparative experimental and clinical study of the efficiency of the combination of dermal matrix with autologous and allogenic cells for creation of an optimal biological wound coating. Experiments on outbred mice have demonstrated that the use of dermal matrix in combination with allogeneic or autologous cells shortens the inflammation phase and accelerated maturation of the granulation tissue. Clinical application of biological wound dressing based on dermal matrix in combination with autologous or allogeneic mesenchymal multipotent stromal cells prevented septic complications and shortened the time of preparing extensive traumatic wounds with skin defect to autodermoplasty.

The article highlights techniques and effects of intraoperative mechanic blood reinfusion in patients with trauma and intraabdominal bleeding in extend, exceeding the self circulating blood volume. The high efficacy of the self blood reinfusion during the emergency operation allowed the 2-fold decrease of the hospital and overall mortality. The mechanic blood reinfusion proved to be a safe and clinically effective method of the globular blood volume deficiency compensation, especially in emergency surgery.

The results of intraoperative hemotransfusion of 112 patients with abdominal injuries, complicated with bleeding and the loss of more then 70% of the circulating blood. The aim of the study was to compare the efficacy of auto- and allohemotransfusion. The first method allowed to decrease both the overall lethality (p=0.039) and postoperative lethality (p=0.018). The odds ratio by the predominate autohemotransfusion was 1.385 (95% 1.101-1.741), where as the odds ratio by the predominate allohemotransfusion was 0.403 (95% 0.183-0.885; p=0.011).

Severe sepsis and septic shock remain the most serious problem of critical care medicine today with a mortality of 25-80%. Bacterial endotoxin is of considerable importance in the pathogenesis of sepsis. A selective hemosorbent in which endotoxin adsorption is accomplished via its binding to a synthetic peptide (LPS-A) has been recently designed in Sweden. The purpose of the study was to evaluate the efficiency of using the LPS-A in the complex treatment of patients with severe sepsis. Eight patients in whom the course of the underlying disease was complicated by the development of gram-negative sepsis in the postoperative period were examined. The patients had an APACHE II score of 20.4 +/- 4.5 LPS-A was performed using the hemosorbent Alteco LPS (Lund, Sweden) and an Aquarius hemoprocessor (Edwards Lifesciences) (U.S.A.). The use of LPS-A caused a significant reduction in endotoxin levels by 2.9 times. The level of procalcitonin was increased in all patients; after a LPS-A session, it was significantly decreased by 1.9 times. The use of LPS-A resulted in diminished tachycardia, elevated blood pressure and could reduce the doses of vasopressor amines. There was an improvement in lung gas exchange parameters. A study of the effect of LPS-A on the immune system showed a significant reduction (36.6%) in neutrophil oxygen-dependent metabolism according to the data of the spontaneous HCT test, a 2.4-fold decrease in large circulating immune complexes, and a rise in the count of mononuclear cells that died both by necrosis and apoptosis, the count of the cells significantly increased by 5.3 times in late apoptosis. A LPS-A session caused a significant drop in the blood level of endotoxin and procalcitonin and it was accompanied by a trend toward better hemodynamics and lung gas exchange and alleviated organ dysfunction. At the same time LPS-A showed the ambiguous effect on immune system parameters, which calls for further study.

Apoptosis of mononuclear cells, dead leukocyte and CD95+ lymphocytes content in venous blood of 15 patients were evaluated by flow cytometry within 2-3 day after diagnosis of sepsis. The number of CD95+ lymphocytes increased in 40% of the cases and that of lymphocytes expressing Fas receptor decreased in 27%. The number of mononuclear cells in venous blood increased 7 times compared with physiological norm; this rise was a reliable predictor of the unfavourable outcome of the disease. Elimination of dead leukocytes by granulocytophoresis produced good therapeutic result.

The study was undertaken to evaluate the immunomodulatory effects of various methods of extracorporeal hemocorrection in septic patients. Thirty-two patients in whom the course of the underlying disease was complicated by the development of sepsis were examined. A package of therapeutic measures involved debridement of an infection focus, de-escalation antibacterial therapy, nutritional support, and immunomodulatory therapy. All the patients underwent procedures of plasma filtration and continuous venovenous hemofiltration; granulocytopheresis was carried out in 5 patients. Granulocytopheresis was shown to eliminate dead leukocytes from the bloodstream within a few hours, which contributed to clinical improvement. Plasmafiltration in patients with sepsis resulted to a reduction in the degree of a systemic inflammatory reaction, to normalization of coagulation homeostasis, which indirectly affected immunological parameters. Continuous venovenous hemofiltration made it possible to make continuous elimination of the low and average molecular-weight substances, corrected liquid balance, and well affected gas exchange in the lung.

Sepsis with 18-to-33% mortality also remains the most serious problem of modem medicine today. Forty-five patients treated at the N. V. Sklifosovsky Research Institute of Emergency Care in June 2006 to February 2008 were examined. Twenty-seven primary blood donors were examined to determine the physiological concentration of dead leukocytes. Two groups of patients were formed according to the signs of a systemic inflammatory reaction (SIR). A study group included 29 patients in whom the course of the underlying disease was complicated by the development of sepsis. All the patients from the study group underwent extracorporeal hemocorrection techniques (EHT): plasma filtration (PF) and continuous venovenous hemofiltration (CVVHF). A granulocytopheresis procedure was performed in 4 patients. A control group comprised 16 patients without clinical signs of SIR. The blood taken from patients with sepsis showed the significant increase in the levels of dead leukocytes as compared with both the physiological normal values and the values obtained in patients without signs of systemic inflammation, which significantly correlates with the severity of organ dysfunction. The elevated content of dead leukocytes is an independent risk factor for a poor outcome. Plasma filtration used in sepsis patients facilitates a significant reduction in the blood concentration of dead leukocytes. The first experience with granulocytopheresis in patients with severe sepsis, a leukemoid reaction and high absolute concentrations of dead leukocytes is indicative of its clinical effectiveness, which is manifested itself by the elimination of dead leukocytes.

Clinical and morphological comparison of wound healing after transplantation of living cultured allofibroblast on days 1-2 after the injury, collagen-1-based dressing with PDGF-BB, and traditional dressing with levomecol ointment showed that bioactive dressing accelerated wound epithelialization (5-7 days vs. 20-22 days with gauze dressing); the incidence of suppurative complications decreased, no crust formed, and epithelialization was not associated with the formation of a hypertrophic cicatrix. Biological dressing based on living cultured allofibroblasts and collagen-1 with PDGF-BB exhibited equal stimulatory effects on burn wound healing.

The biologic band is created on the base of type I collagen and platelet derived growth factor. It was applied for extensive burns of IIIa grade at 1-2nd day after a trauma, which led to epidermis reconstruction at 5-7th day. Authors assume that the effect of the band is determined by the time of its use--1-2nd day, when stimulating exudation qualities combine with healing properties of the bandage. The effectiveness of the collagen band showed to be equal to those of fibroblast transplantation.

Acute purulent mediastinitis (APM) is one of the most complicated forms of surgical infection, showing a high incidence of sepsis--from 45 to 100%, mortality rates of 17 to 80%. Sixty-eight patients with APM admitted to the N. V. Sklifosovsky Research Institute of Emergency Care in October 2002 to March 2007 were examined. Postoperatively, all the patients received extracorporeal hemocorrection techniques (EHT): plasmapheresis (PA) via filtration and continuous venovenous hemofiltration (CVVHF). According to the time of initiation of EHT, the patients were divided into 2 groups: 1) 34 patients in whom EHT was initiated within the first 24 hours after surgery; 2) 34 patients in whom it was started on postoperative day 2. The efficiency of early use of EHT in the complex therapy of APM was evaluated. The early initiation of EHT (PA and CVVHF) caused a reduction in endogenous intoxication and the magnitude of a systemic inflammatory reaction, which resulted in the rapidest restoration of the size of major populations and subpopulations of lymphocytes and prevented the development of immune system incompetence. The early use of EHT caused a significant reduction in hospital mortality (11.8 and 35.3% in the early and late EHT use groups, respectively).

Results of immunological examination of 423 patients with syndrome of systemic inflammatory reaction of different ethiology (mediastinitis--78 patients, peritonitis--85, severe acute pancreatitis--91, trauma of thorax and abdomen complicated with massive hemorrhage--111, severe burn trauma--40 patients) are analyzed. For complex and objective assessment of immune reaction the system of scores has been developed. This scale permitted to detect the immune disorders, to diagnose the degree of immune insufficiency at early stages of disease, to predict the purulent complications and to start timely immune therapy.

The results of the immunological study of blood in 89 patients with severe acute pancreatitis (SAP) were analyzed. Immune reactions in SAP were characterized by the presence of moderate leukocytosis, an increase in the phagocytic and metabolic activity of neutrophils, relative lymphopenia, a decrease in the number of B-lymphocytes. The average values of the parameters of the immunogram of patients with the favorable course of the disease on days 2 - 3 were analogous to the average population values in 89 examinees. The direction and the degree of manifestation were slightly variable and could be registered within one standard quadratic deviation. Such heterogeneity in the character of immune reactions made it possible to regard the average values of the immunogram parameters of these patients as "the norm". For the objective evaluation of the immune status of SAP patients a scale for the evaluation, in points, of disturbances in immune reactions to the destructive process in the pancreas was developed. This scale for the evaluation, in points, of the severity of disturbances in the immune status of SAP patients made it possible to prognosticate the development of complications.

An analysis of data of immunological investigation of blood of 89 patients with a severe acute pancreatitis (SAP) on the 2-3 day of the disease has revealed changes in immunogram characteristic also of other pathological conditions and pointing to a systemic inflammatory reaction. Among the patients who did not have purulent complications against the background of therapy of SAP, the character and degree of changes in immunogram were in line with average values, and so they could be considered as "a norm of pathology". A system of scores of impairments of the immune response to the destructive process was developed for an objective evaluation of the immune status of patients with SAP. In patients with a normal physiological immune response (sum of scores 0-5) the prognosis of the disease is considered to be good. No immunocorrectors should be included in the complex of treatment measures. In patients with an inadequate pathological immune response (sum of scores 9 and more) the prognosis is unfavorable. The complex of treatment measures for such patients should include immunocorrectors.

The subjects of the study were 82 patients with penetrating gunshot (17 cases) and stab-and-cut (65 cases) thoracal and abdominal wounds, complicated by an acute blood loss of 600 to 4500 ml. Aerobic metabolism of neutrophiles was studied using spontaneous and activated HCT test and chemoluminescent analysis; the concentration of acute phase proteins (fibronectin, ceruloplasmin, C-reactive protein, and haptoglobin) during the first two days after surgery was measured. The study revealed that bigger blood loss was associated with more pronounced increase of the number of spontaneously activated neutrophiles with low bactericidal potential, decrease of fibronectin and ceruloplasmin concentrations, and increase of C-reactive protein and haptoglobin concentrations. Retrospective analysis of the treatment results showed that the rate of suppurative complications did not depend on the wound character or blood loss volume, but was higher in patients with prolonged disbalance of acute phase proteins and aerobic metabolism (12.4% vs. 2.6% in patients without such disturbances).

In 62 patients with acute poisoning, blood gases, immunograms, hemorheology, oxidative stress, and the blood levels of biogenic amines were studied at the toxicogenic and somatogenic stages of poisoning. It was found that the development of toxicohypoxic encephalopathy was affected by the magnitude of changes in laboratory homeostatic parameters among which hemorheological and immunological were prominent.

The inhibiting activity of blood sera obtained from 20 volunteers immunized with the multi-component vaccine "PYOPOL", 25 nonimmune donors and 7 lots of human antistaphylococcal immunoglobulin with respect to Staphylococcus aureus, Escherichia coli, Proteus vulgaris and Pseudomonas aeruginosa has been studied in the liquid nutrient medium with the use an automated microbiological analyzer. The sera of donors immunized with the vaccine "PYOPOL" have been found to possess high antimicrobial activity, comparable with the definite concentration of the antiseptic agent "Myramistin". The proposed method may be recommended for the evaluation of the bacteriostatic activity of different immuno- biological preparations.

To develop a quantitative method of evaluation of the volume and degree of intraoperative blood loss.Blood loss quantitation is based on maximal collection of the lost blood and its measurement by lost globular volume (GV), i.e. erythrocyte containing medium with Ht 1.0.The total of the components of lost GV was termed estimated blood loss (EBL). Its amount was calculated by GV deficiency from the due in the patient and volume of lost blood. The method was tried in surgical and traumatological practice in 317 operated patients (multitrauma--65, hip bone fractures -98, uterine myoma -105, aneurysm of abdominal aortic part -49).The volume of blood loss guided the physician to reestablish volume of circulating blood--the volume of infused blood exceeded EBL 1.3-1.7-fold. GV deficiency (in %) showed severity of blood loss and gave approaches to its adequate compensation.

The publications on the etiology, bacteriological and immunochemical diagnostics of pneumonia, as well as on the role of immunotherapy and prophylaxis in the treatment of this disease, are analyzed. The importance of immunological methods in the diagnostics of pneumonia is pointed out. Approaches to the immunotherapy and immunoprophylaxis of pneumonia are updated.

Results of combined treatment of 314 patients with acute pancreatitis, including 58 (15.1%) with pancreonecrosis were analyzed. Etiologic factors of acute pancreatitis were alcohol (59% patients), diseases of the bile ducts (31.5%), surgery (2.5%). Up-to-date diagnostic criteria of severe pancreatitis are presented, character of complications is analyzed. Treatment policy in acute edematous pancreatitis was conservative. In calculous cholecystitis cholecystectomy was performed after regress of acute pancreatitis. Fermentative ascitis-peritonitis was the indication for laparoscopy in aseptic phase of pancreonecrosis. US- and CT-guided puncture and drainage were often used. Surgeries were performed only for complications of pancreonecrosis, more often through mini-approaches. General lethality in acute pancreatitis was 1.9%, in pancreonecrosis - 10.7%, postoperative lethality in pancreonecrosis was 16.6%.

Clinical and laboratory estimation of the efficacy of pectins in complex treatment of patients with intracranial hematoma was performed. It was shown that in the group of the patients treated with pectins vs the control group development of pyo-inflammatory infections was less frequent, the indices of the immunity status improved and a more rapid decrease of the intoxication and a more rapid normalization of the composition of various biotopes in the patients were observed.

The influence of the newly developed complex vaccine Pyopol, containing the antigens of opportunistic microorganisms and polyoxydonium used as immunomodulator, on the oxygen-dependent metabolism of neutrophils was studied. The study revealed that the main components of the vaccine, both individually and in association, did not change cellular activity in the range of concentrations used in this study. The inhibition of the oxygen-dependent metabolism of neutrophil granulocytes in the presence of native or weakly diluted vaccine occurred due to the cytotoxic effect of thimerosal used as preservative.

The comparative study of the relationship between the levels of serum antibodies to the antigens of opportunistic microorganisms of 5 genera (Pseudomonas, Proteus, Staphylococcus, Klebsiella, Escherichia) and the microbial status was carried out. A total of 854 patients from 10 profile departments of a surgical hospital were examined. Population analysis and statistical methods of processing the results of the examination of 353 practically healthy subjects and 268 blood and plasma donors permitted the norms for the levels of specific antimicrobial antibodies (decreased, normal, elevated levels) were established. The constancy coefficients of Pseudomonas, Staphylococcus, Klebsiella, Escherichia and the number of patients with elevated titers of specific serum antibodies were found to be positively correlated (r = +0.47-0.89, p0.01). The data thus obtained made it possible to substantiate the importance of population serological investigations for the evaluation of the epidemiological situation in the surgical hospital.

Clinical and laboratory evaluation of pectins application efficacy demonstrated that the patients treated with pectines has lower frequency of bacteriemia, intoxication, infectious complications and lethality. Pectins use per os resulted by acceleration of burns wounds healing and by lower microbial dissemination. Frequency of coagulase-positive staphylococci, streptococci and enterococci isolation was also lower in the treated group of patients when compared to the control group. Feces microbial profile also demonstrated positive trend--normal microflora (bifidobacteria and lactobacilli) enhanced, opportunistic bacteria diminished. Pectins application provided normalization of leucocytes, lymphocytes, B-lymphocytes, immunoglobulins A and G. The results of investigation demonstrated pectins efficacy at the complex treatment regimes of the patients with burns.

The mechanisms of action of hyperbaric oxygenation (HBO) in toxic hypoxic encephalopathy (THE) were studied using clinical psychopathological examinations, functional and laboratory tests in 268 patients with THE treated by a therapeutic complex including HBO and 75 patients with THE treated routinely (controls). The earliest possible addition of HBO to a complex of treatment of THE patients led to involution of signs of brain edema shown by computer tomography (CT) and improvement of its functional activity, while in patients exposed to HBO later the psychoorganic symptoms and CT signs of cerebral ischemia did not disappear. Detoxifying, neuroimmunomodulating, and neuroimmunostimulating effects of HBO in THE were demonstrated. Early HBO treatment decreased the dysfunction of various compartments of the brain characteristic of THE. The technology of HBO developed by the authors prevented the development of socially dysadapting psychoneurological disorders and reduced the mortality of THE patients.

Results of immune status examination on day 1-2 and 4-8 after operation in 217 patients with penetrating gunshot and stab-incised wounds of thorax and abdomen were analyzed. Correlation between immunogram and volume of acute hemorrhage was studied. Types of immunograms characteristic of good and complicated postoperative period were determined, their frequency was estimated, that permits to perform substantiated and timely special correction for acceleration of immunorehabilitation and prevention of infectious complications.