Modulating the Biologic Activity of Mesenteric Lymph after Traumatic Shock Decreases Systemic Inflammation and End Organ Injury.

INTRODUCTION: Trauma/hemorrhagic shock (T/HS) causes the release of pro-inflammatory mediators into the mesenteric lymph (ML), triggering a systemic inflammatory response and acute lung injury (ALI). Direct and pharmacologic vagal nerve stimulation prevents gut barrier failure and alters the biologic activity of ML after injury. We hypothesize that treatment with a pharmacologic vagal agonist after T/HS would attenuate the biologic activity of ML and prevent ALI. METHODS: ML was collected from male Sprague-Dawley rats after T/HS, trauma-sham shock (T/SS) or T/HS with administration of the pharmacologic vagal agonist CPSI-121. ML samples from each experimental group were injected into naïve mice to assess biologic activity. Blood samples were analyzed for changes in STAT3 phosphorylation (pSTAT3). Lung injury was characterized by histology, permeability and immune cell recruitment. RESULTS: T/HS lymph injected in naïve mice caused a systemic inflammatory response characterized by hypotension and increased circulating monocyte pSTAT3 activity. Injection of T/HS lymph also resulted in ALI, confirmed by histology, lung permeability and increased recruitment of pulmonary macrophages and neutrophils to lung parenchyma. CPSI-121 attenuated T/HS lymph-induced systemic inflammatory response and ALI with stable hemodynamics and similar monocyte pSTAT3 levels, lung histology, lung permeability and lung immune cell recruitment compared to animals injected with lymph from T/SS. CONCLUSION: Treatment with CPSI-121 after T/HS attenuated the biologic activity of the ML and decreased ALI. Given the superior clinical feasibility of utilizing a pharmacologic approach to vagal nerve stimulation, CPSI-121 is a potential treatment strategy to limit end organ dysfunction after injury.

Sirtuin 1 Agonist Minimizes Injury and Improves the Immune Response Following Traumatic Shock.

Survival from traumatic injury requires a coordinated and controlled inflammatory and immune response. Mitochondrial and metabolic responses to stress have been shown to play a role in these inflammatory and immune responses. We hypothesized that increases in mitochondrial biogenesis via a sirtuin 1 agonist would decrease tissue injury and partially ameliorate the immunosuppression seen following trauma. C57Bl/6 mice were subjected to a multiple trauma model. Mice were pretreated with either 100 mg/kg per day of the sirtuin 1 agonist, Srt1720, via oral gavage for 2 days prior to trauma and extended until the day the animals were killed, or they were pretreated with peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α) siRNA via hydrodynamic tail vein injection 48 h prior to trauma. Markers for mitochondrial function and biogenesis were measured in addition to splenocyte proliferative capacity and bacterial clearance. Srt1720 was noted to improve mitochondrial biogenesis, mitochondrial function, and complex IV activity following traumatic injury (P < 0.05), whereas knockdown of PGC1α resulted in exacerbation of mitochondrial dysfunction (P < 0.05). These changes in mitochondrial function were associated with altered severity of hepatic injury with significant reductions in serum alanine aminotransferase levels seen in mice treated with srt1720. Splenocyte proliferative capacity and intraperitoneal bacterial clearance were evaluated as markers for overall immune function following trauma-hemorrhage. Treatment with Srt1720 minimized the trauma-induced decreases in splenocyte proliferation (P < 0.05), whereas treatment with PGC1α siRNA led to diminished bacterial clearance. The PGC1α signaling pathway is an important regulator of mitochondrial function and biogenesis, which can potentially be harnessed to protect against hepatic injury and minimize the immunosuppression that is seen following trauma-hemorrhage.

Early trauma-hemorrhage-induced splenic and thymic apoptosis is gut-mediated and toll-like receptor 4-dependent.

Immune depression after trauma-hemorrhage has been implicated as an important factor in the pathogenesis of sepsis and septic-organ failure. Although recent studies have implicated immune-cell apoptosis as an important factor in the evolution of this posttrauma immune-suppressed state, neither the initial triggers that induce this response nor the cellular pathways through which these triggering pathways act have been fully defined. Thus, the current study tests the hypothesis that acute splenic and thymic immune-cell apoptosis developing after trauma-hemorrhagic shock (T/HS) is due to gut-derived factors carried in intestinal lymph and that this T/HS lymph-induced immune depressed state is mediated through Toll-like receptor 4 (TLR4). The first set of experiments documented that T/HS caused both thymic and splenic immune-cell apoptosis as measured by TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) and caspase-3 immunohistochemistry and that this increase in apoptosis was totally abrogated by mesenteric lymph duct ligation. In subsequent experiments, mesenteric lymph collected from animals subjected to T/HS or trauma-sham shock were injected into TLR4-deficient (TLR4mut) mice or their wild-type (WT) littermates. Trauma-hemorrhagic shock, but not trauma-sham shock, lymph caused splenic apoptosis in the WT mice. However, the TLR4mut mice were resistant to T/HS lymph-induced splenic apoptosis. Furthermore, the WT, but not the TLR4mut mice developed splenic apoptosis after actual T/HS. In conclusion, gut-derived factors appear to initiate a sequence of events that leads to an acute increase in splenic and thymic immune-cell apoptosis, and this process is TLR4-dependent.

Rons formation under restrictive reperfusion does not affect organ dysfunction early after hemorrhage and trauma.

Reactive oxygen species have been implicated in the pathophysiology of early reperfusion. We aimed to determine 1) reactive oxygen and nitrogen species (RONS) formation in organs of rats and 2) its pathophysiological relevance during a phase of restrictive reperfusion after hemorrhagic/traumatic shock (HTS). Fifty-seven male Sprague-Dawley rats were subjected to a clinically relevant HTS model, featuring laparotomy, bleeding, and a phase of restrictive reperfusion. The RONS scavenger 1-hydroxy-3-carboxy-2,2,5,5-tetramethyl-pyrrolidine hydrochloride (continuous i.v. infusion) and electron paramagnetic resonance spectroscopy were applied for RONS (primarily superoxide and peroxynitrite) detection. Compared with sham-operated animals, the organ-specific distribution of RONS changed during restrictive reperfusion after HTS. Reactive oxygen and nitrogen species formation increased during restrictive reperfusion in red blood cells and ileum only but decreased in the kidney and remained unchanged in other organs. Hemorrhagic traumatic shock followed by restrictive reperfusion resulted in metabolic acidosis, dysfunction of liver and kidney, and increased oxidative burst capacity in circulating cells. Plasma RONS correlated with shock severity and organ dysfunction. However, RONS scavenging neither affected organ dysfunction nor oxidative burst capacity nor myeloperoxidase activity in lung when compared with the shock controls. In summary, a phase of restrictive reperfusion does not increase RONS formation in most organs except in intestine and red blood cells. Moreover, scavenging of RONS does not affect the early organ dysfunction manifested at the end of a phase of restrictive reperfusion.

Mechanism of the salutary effects of estrogen on kupffer cell phagocytic capacity following trauma-hemorrhage: pivotal role of Akt activation.

Kupffer cells are macrophages in the liver whose major role is to clear circulating pathogens. Decreased phagocytic capacity of Kupffer cells may result in severe systemic infection. We tested the hypothesis that the depressed Kupffer cell phagocytic capacity following trauma-hemorrhage is enhanced by estrogen administration and this occurs due to maintenance of Fc receptor expression and cellular ATP content via the activation of Akt. Male C3H/HeN mice were subjected to sham operation or trauma-hemorrhage and sacrificed 2 h thereafter. Estrogen, with or without an estrogen receptor antagonist (ICI 182,780), a PI3K inhibitor (Wortmannin), or vehicle, was injected during resuscitation. Kupffer cell phagocytic capacity was tested in vivo. The expression of Fc receptors, of Akt phosphorylation, of p38 MAPK phosphorylation, of DNA binding activity of NF-kappaB and ATP content of Kupffer cells were also determined. Trauma-hemorrhage suppressed Kupffer cell phagocytosis by decreasing Fc receptor expression and Akt activation; however, it induced p38 MAPK activation and increased NF-kappaB activity. Cellular ATP levels were also decreased following trauma-hemorrhage. Administration of estrogen following trauma-hemorrhage increased phospho-Akt levels and normalized all the parameters described as well as plasma levels of TNF-alpha, IL-6, and IL-10. Coadministration of ICI 182,780 or Wortmannin abolished the beneficial effects of estrogen in improving the phagocytic capacity of Kupffer cells following trauma-hemorrhage. Thus, activation of Akt plays a crucial role in mediating the salutary effect of estrogen in restoring trauma-hemorrhage-induced suppression of Kupffer cell phagocytosis.

[Use of acetylcysteine and polyoxydonium for the correction of immune homeostasis and antioxidant system impaired by combined action of 1,2-dichloroethane and heavy mechanical trauma].

The experiments on noninbred rats showed that a combination of heavy mechanical trauma and 1,2-dichloroethane in a single dose of 0.5 LD50 leads to summation of the immunosuppressive and prooxidizing effects of both factors. A combined administration of acetylcysteine and polyoxidonium on the background of the combined action of a heavy mechanical trauma and 1, 2-dichloroethane (1.5 LD50) reduces the suppression of immune responses and the activation of lipid peroxidation caused by these factors.

[Effect of Soluvit on stress reaction and leucocyte function in serious burn patients in shock stage].

OBJECTIVE: To investigate the effect of Soluvit on stress reaction and leucocyte function in serious burn patients in shock stage. METHODS: Eighty-seven serious burn patients, who did not undergo operation, were divided into Soluvit treatment group and control group randomly. Patients in Soluvit treatment group were treated with two bottles of Soluvit everyday from postburn 1 st to 14 th day. Patients in control group were given 500 ml normal saline infusion instead. Blood samples were collected for determination of cortisol and malondialdehyde every 4 hours on postburn 1 st day. Leucocyte were isolated for testing chemotaxis distance and phagocytic power on postburn 7 th and 14 th day respectively. RESULTS: The serum cortisol contents in serious burn patients were significantly elevated at 2-4 hours after treatment in Soluvit treatment and control groups. Serum cortisol and malondialdehyde levels were higher than normal values in all serious burn patients in shock stage at 10-12 hours after treatment. But the changes of serum cortisol and malondialdehyde in Soluvit treatment group were all lower than those in control group at 10-48 and 14-48 hours after treatment, respectively (all P<0.05). Leucocyte chemotaxis distance in Soluvit treatment group was longer than that in control group on both postburn 7 th and 14 th day (P<0.05 and P<0.01). In contrast, there were no significant differences in phagocytic power between two groups (both P>0.05). CONCLUSION: Above results suggest that Soluvit can mitigate stress reaction and lipid peroxidation action, but enhance leucocyte chemotaxis function in severe burn patients.

Gut-lymph hypothesis of systemic inflammatory response syndrome/multiple-organ dysfunction syndrome: validating studies in a porcine model.

BACKGROUND: Trauma-hemorrhagic shock (T/HS) mesenteric lymph from rats has multiple biological properties and appears to cause organ injury via the activation of neutrophils and endothelial cells. As the next step in testing the potential clinical relevance of these rodent studies, we utilized a swine T/HS model to determine whether the intestinal lymph results observed in the rodent could be replicated in swine. A porcine model was chosen because the pig and human cardiovascular and gastrointestinal physiology are similar. METHODS: Male pigs were subjected to T/HS and a major intestinal lymph duct was cannulated. Hemorrhagic shock (mean arterial pressure, 40 mm Hg) was performed by withdrawing blood, for 3 hours or until the base deficit reached -5. Animals were then resuscitated in two stages to mimic the prehospital and hospital phases of resuscitation. Mesenteric lymph was collected hourly throughout the experiment and its biological activity was tested on neutrophils (respiratory burst) and endothelial cells (monolayer permeability and cytotoxicity). RESULTS: T/HS lymph but not trauma-sham shock lymph (T/SS) increased neutrophil activation as reflected by an augmented respiratory burst. Likewise T/HS lymph collected at all time points up to 5 hours postshock significantly increased endothelial cell permeability by twofold or greater (p < 0.05), whereas T/HS lymph produced during the first 2 hours postshock was cytotoxic for endothelial cells (viability 70%, p < 0.05 vs. preshock). In contrast, T/SS lymph had no effect on the endothelial cells. CONCLUSION: This large animal model validates rodent studies showing that the shock-injured gut releases biologically active factors into the mesenteric lymph and these factors activate neutrophils and injure endothelial cells.

[Influence on the concentration of plasma endotoxin by inhibition of complement activation in traumatic hemorrhagic shock rats].

OBJECTIVE: To investigate the influence on the concentration of plasma endotoxin by inhibition of complement activation in traumatic hemorrhagic shock rats. METHODS: Eighty male SD rats were randomly divided into two groups: control and cobra venom factor (CVF) treatment groups. The hemorrhagic shock induced by trauma was replicated in both groups. The animals were killed preshock and at 1, 6, and 24 hours postresuscitation. Twenty-four hours before hemorrhage, rats were given a mainline dose of either 50 microg/kg CVF or an equal volume of saline solution. The plasma and serum samples were collected at each time point to determine the concentration of endotoxin, the activity of CH50 and diamine oxidase (DAO), and the level of tumor necrosis factor (TNF-alpha) at various time points in two groups. RESULTS: Compared with preshock in control group, serum CH50 levels were decreased promptly at 1 hour postresuscitation. Markedly elevation of the levels of endotoxin and TNF-alpha in blood were found at early time after resuscitation, and they were come rapidly back to the basic level at 6 and 24 hours phase. The activity of DAO in blood was increased significantly at 1 and 6 hours after resuscitation and declined promptly at 24 hours. Compared with the control group, significantly decline of the levels of endotoxin, TNF-alpha and DAO at the various time points after resuscitation were also found in the CVF group. The levels of CH50 in CVF group were always less than 5% during the experiment. CONCLUSION: In traumatic hemorrhagic shock rats CVF pretreatment could decline plasma endotoxin levels by preventing the injury of intestine and gut barrier function, decrease endotoxin translocation and reduce plasma endotoxin levels.

[Influence of the escharectomy during stock stage on the peripheral lymphocyte apoptosis and the antigen presentation function of monocytes in peripheral blood of scalded rats].

OBJECTIVE: To investigate the influence of escharectomy at different time-points after burn injury on the lymphocyte apoptosis and the antigen presentation function of monocytes in peripheral blood of scalded rats. METHODS: One hundred and thirty-six Wistar rats were randomly divided into normal control ( C,n = 8 ), scald ( S, n = 64,without treatment after scald) , A ( n = 40, with escharectomy at 36 post-burn hour( PSH) ) , B ( n = 24, with escharectomy at 72 PSH ) groups. The rats in A , B, S groups were inflicted with 30% TBSA full-thickness scald. The rats in S group were sacrificed on 6,12,24,72,120,168,216, 288 PSH, while those in A and B groups were sacrificed at 72 -288 PSH, 168 -288PSH, respectively. The rats in C group were also sacrificed as control. The apoptotic rate of peripheral lymphocytes, the positive expression rate of MHC- II in mononuclear cells, the changes in concentration of IL-4 and gamma-IFN were determined in each group. The correlation of above indices were also analyzed. RESULTS: (1) The apoptotic rate of peripheral lymphocyte in S group were increased dramatically at 6PSH, peaking at 24 PSH( 18. 19+/-1.42% ) , then decreasing gradually, reaching the lowest level at 72 PSH(8. 25+/-0.56% ) , then it increased gradually again, approaching almost the peak value at 288 PSH( 17.81 +/- 1.99% ). The values were all obviously higher than those in C group( P <0.05). The apoptotic rates of peripheral lymphocyte in A and B groups were evidently lower than that in S group ( P <0. 01). (2) The positive expression rate of MHC-II in monocyte was decreased sharply at 6 PSH, and it was 20% lower than that in C group (37. 2 +/- 2. 4% ) at 24 PSH. It then increased gradually, but it was significantly lower than that in A, B groups at 288 PSH (18. 8 +/-2. 8, P <0.01). (3) The plasma level of y-IFN in S group increased gradually from 6 PSH on, peaking at 24 PSH(440. 8 +/-25. 1 )ng/L,then decreasing gradually , and it reached the lowest level at 288 PSH (51.3 +/-37.0) ng/L. The IL-4 level in S group was increased gradually ,peaking at 288 PSH (78. 1+/-2. 8) ng/L. (4) There was negative correlation between the expression rate of MHC- II in S group and IL-4/gamma-IFN ratio in escharectomy groups during 72 - 288 PSH ( r = - 0. 96, P < 0. 05). CONCLUSION: Eacharectomy after scald can inhibit peripheral lymphocyte apoptosis, slow down the insertional tendency of IL-4/gamma-IFN , and ameliorate the antigen presentation function of monocytes. Moreover, escharectomy during shock stage can markedly promote the immune function of monocytes.

[The change of the erythrocyte chemokine receptor binding activity in the shock stage of burn rats].

AIM: To investigate the change of the erythrocyte chemokine receptor(ECKR) binding activity in the shock stage of burn rats. METHODS: SD rats were randomly divided into two group, burn and control groups. In the burn group rats, 30% total body surface area (TBSA)were scalded to III degree. The binding activity of rat ECKR in the shock stage was detected by ELISA using IL-8 as ligand at various time points (0.5, 2, 4, 8, 16, 24 and 48 hours) after burn. RESULTS: Compared with control group, the binding activity of rat ECKR declined significantly half an hour after burn and maintained at low level for 48 hours (P<0.01). The comparison of the binding activity of rat ECKR at various time points after burn indicated that the binding activity declined gradually from 2 hours (P<0.01-0.05), reached lowest value 24 hrs, and then rose significantly 48 hrs after burn (P<0.01). CONCLUSION: The ECKR binding activity declined significantly after burn, suggesting erythrocytes may participate in the regulation of chemokines and play some role in the inflammation.

Burn injury initiates a shift in superantigen-induced T cell responses and host survival.

Severe injury induces a temporal shift in immune reactivity that can cause serious complications or even death. We previously reported that mice exposed to bacterial superantigen (SAg) early after injury undergo a strong SAg response with lethal consequences. This study compares the early and late effects of burn injury on SAg reactivity in vivo to establish how injury influences adaptive immune responses. We found that mice challenged with ordinarily sublethal doses of staphylococcal enterotoxin A or staphylococcal enterotoxin B at 1 day after burn injury exhibited high mortality, whereas no mortality occurred at 7 days after injury. This shift in mortality correlated with higher Th2-type cytokines (IL-4 and IL-10) being expressed by CD4(+) and CD8(+) T cells from burn as opposed to sham mice at 7 days after injury. Lymph node cells from burn-injured mice also produced higher levels of Th2-type cytokines at 7 days after injury. The results of cell-mixing studies using CD4(+) and CD8(+) T cells mixed with APCs from sham or burn mice suggested that changes in both T cells and APCs are involved in the altered SAg response. Finally, the biological significance of altered SAg reactivity following injury was shown by demonstrating that blocking IL-10 activity in vivo caused higher SAg-induced mortality at 7 days after injury. These findings support the idea that injury promotes a Th2-type shift in adaptive immune reactivity. Although prior studies link this counterinflammatory-type response to lowered resistance to infection, the present results suggest it may sometimes benefit the injured host.

[Influence of escharectomy during shock stage on the systemic and intestinal immune function in scalded rats].

OBJECTIVE: To investigate the influence of escharectomy during shock stage on systemic and intestinal immune function and its mechanism in scalded rats. METHODS: Ninety-six Wistar rats were employed in the study of which 8 were used as normal control group. The donor skin from the trunk in twenty-four rats were preserved in liquid nitrogen. The other 64 rats were subjected to 30% full-thickness scalding, and they were randomly divided into A (n = 24, no treatment after scalding), B (n = 24) and C (n = 16) groups. Physiological saline was intraperitoneally injected (50 ml/kg) on the 24 post-scalding hours to the rats in the B and C groups. The rats in B group underwent escharectomy during shock stage, and the excision wounds were covered with the cryo-preserved alloskin. The rats in C group received the same treatment as in B group but at 72 post-scalding hours. The change in the proliferative ability of splenic lymphocytes, the plasma and intestinal tissue content of interleukin 2 (IL-2), the contents of sIgA in intestinal mucus, and the content of DAO in the intestinal tissue were observed on 2, 4 and 8 post burn days (PBD) in A and B groups and also on 4 and 8 PBD in C group, respectively. RESULTS: The splenocytic proliferative ability, IL-2 level in the plasma and intestinal tissue, and the sIgA content in intestinal mucus in the rats in A, B and C groups were lower than that in control group at all time points (P < 0.05). The proliferative ability of splenic lymphocytes in B group on 4 and 8 PBD and in C group on 8 PBD respectively was similar to that in control group. Whereas the IL-2 content in plasma and in intestinal tissue was higher in B and C groups than that in A group (P < 0.01). The sIgA content in intestinal mucus in B group was twice of that in C group respectively [(3.51 +/- 2.14) mg/g vs (1.40 +/- 0.64) mg/g, (3.03 +/- 0.95) mg/g vs (1.52 +/- 1.26) mg/g (P < 0.05 or P < 0.01)] on 4 and 8 PBD. The DAO activity in the intestinal tissue in A group was lower than that in control and B group (P < 0.05) on 4 and 8 PBD. CONCLUSION: Escharectomy during shock stage might be beneficial to the recovery of the systemic and intestinal immune functions in rats with scalding injury.

[Escharectomy during burn shock on Th1/Th2 polarization of helper T lymphocytes in rats after thermal injury].

OBJECTIVE: To determine the type-1/type-2 cytokine response of helper T lymphocytes after thermal injury, and to investigate the effects of escharectomy during burn shock stage on the polarization of Th cells. METHODS: One hundred and sixty male Wistar rats were randomized into four groups. In group A, animals were subjected to a 30 percent full-thickness thermal injury without escharectomy. In groups B, C and D, escharectomy and skin allograft were done at 8, 24, and 96 hours postburn, respectively. At 4, 12, 24, 48, 96, 120 and 168 hours postburn, animals were killed and bloods samples as well as spleens were harvested. Enzyme linked immunoadsorbent assay (ELISA) was applied to determine interferon-gamma(IFN-gamma) and interleukin-4(IL-4) levels in blood and spleen tissues. RESULTS: Levels of IFN-gamma and IL-4 rapidly and significantly were increased after scald injury. IFN-gamma levels began to rise within 4 hours postburn, peaking at 24 hours later. IL-4 showed a persistent elevation up to 168 hours postburn, thereby leading to a dominant tendency of Th2 cytokine response on postburn day 7. In group A, all above parameters revealed most obvious changes compared with controls, then ranked in group D, B and C. CONCLUSION: Escharectomy during burn shock stage is helpful to prevent the shift to Th2 cell response after severe thermal injury.

Post-traumatic osteomyelitis: analysis of inflammatory cells recruited into the site of infection.

Device-associated infections after implants or endoprostheses inflict local inflammation and ultimately osteolysis, a clinical entity referred to as posttraumatic osteomyelitis. The underlying molecular mechanisms are not yet known; formation of bacterial biofilms on the implant is presumed, conferring resistance to antibiotics and to host defense mechanisms as well. To gain insight into the pathogenesis of post-traumatic osteomyelitis, the infected site was analyzed for the presence of immunocompetent cells. In 18 patients, the infected site was rinsed intraoperatively. This so-called lavage contained 1-2 x 107 leukocytes, predominantly highly activated polymorphonuclear neutrophils (PMNs), as characterized by low expression of CD62L (selectin), and high expression of the adhesion protein CD18, of the high-affinity immunoglobulin (IgG) receptor CD64, and of the LPS-receptor CD14. CD16, the low-affinity IgG receptor, was affected in some patients only. Because the majority of infections were caused by staphylococci species, the effect of bacteria-derived lipoteichoic acid on PMN of healthy donors was tested in vitro. A similar activation pattern was found: rapid down-regulation of CD62L, a slower loss of CD16, and upregulation of CD18, CD64, and CD14. Lipoteichoic acid signaling required p38 mitogen-activated protein kinase and resulted in induction of CD14-specific mRNA and de novo protein synthesis. We conclude that PMNs infiltrate the infected site, but despite local activation they are unable to clear the bacteria, presumably because of biofilm formation. Our data are consistent with the hypothesis that during the ineffective "frustrated" attempt to phagocytose, PMNs release cytotoxic and proteolytic entities that in turn contribute to the progression of tissue injury and ultimately to osteolysis.

The gene expression of cytokines and chemokines induced by tourniquet shock in mice.

Traumatic shock is one of the major fields in forensic pathology, but its mechanism remains elusive from the pathophysiological aspects. Tourniquet shock has been established as one of the animal models of traumatic shock, and we examined the gene expression of cytokines and chemokines in the lung and liver in tourniquet shock using mice. Tourniquet was conducted by the application of elastic bands with five turns at both the thighs as high as possible for 2 h, followed by reperfusion. In this procedure, more than 90% mice died within 48 h after reperfusion. Serum hepatic transaminase and hematocrit values significantly increased at 2 h after reperfusion, and their elevation was still evident after 10 h. Histopathologically, hemorrhages, congestion and leukocyte recruitment were observed in the lung and liver specimens after 6 h of reperfusion. Immunohistochemical analysis with anti-myeloperoxidase antibody demonstrated a massive neutrophil infiltration in the lung and liver at 2 h or more after reperfusion. RT-PCR analyses demonstrated that the gene expression of interleukin-1beta, tumor necrosis factor-alpha, monocytes chemoattractant protein-1, macrophage inflammatory protein (MIP)-1alpha, MIP-2, KC and vascular endothelial adhesion molecule-1 was most enhanced in the lung and liver at 2 h after reperfusion. Thus, the gene expression of cytokines and chemokines is presumed to be closely related with the onset of tourniquet shock. From the forensic aspects, these cytokines and chemokines are considered to be useful markers for the early diagnosis of tourniquet shock.

[Use of immunomodulator derinat in the treatment of patients with surgical sepsis in traumatic shock]. / Primenenie derinata v lechenii bol'nykh s khirurgicheskim sepsisom pri travme.

The reparative effects of Derinat were studied in 15 patients with traumatic shock whose state was complicated by the development of sepsis. The immunomodulating effect of the drug lies in increasing the number of lymphocytes, reducing the total number of granulocytes and in elevation of the number of functionally valuable cells in them. Using Derinat was shown to facilitate the restoration of the initially reduced number of blood erythrocytes. The maximum effect of the drug was observed within 3-4 days. Readministrations of Derinat should be made twice a week.

[Immunopathogenesis of severe wounds and traumas: possibilities of immune correction]. / Immunopatogenez tiazhelykh ranenii i travm: vozmozhnosti immunokorrektsii.

The authors describe the present-day views on the nature of immune dysfunctions in severe traumas. Based on personal clinical experiences and literature data the authors discuss the role of immune dysfunctions in pathogenesis of the traumatic disease. Special attention is given to the role of the immune system in the development of the life-threatening condition: polyorganic insufficiency whose formation mainly results from disorganization and functional failure of the system of immune reactivity. Clinical investigations have shown high effectiveness of early administration for severe wounds and traumas of a new means of immunocorrection--yeast recombinant interleukin-2 of man (preparation Roncoleukin). The administration of this immunocorrector in complex schemes of intensive therapy of the victims was shown to prevent the development of severe pyo-septic pathology and perfectly change the course of the traumatic disease.