Objective To compare the phenotype of myeloid derived suppressor cells (MDSCs) separated from the bone marrow of mice 3 d and 7 d after cecal ligation and puncture ( CLP) and to elucidate its potential role in the accumulation and immuno-function of MDSCs by determining the expression of microRNA-146a(miR-146a)in order to explore the effect of miR-146 a on immonosuppression of MDSCs in sepsis .Methods A septic model was prepareol by CLP in adult male C57BL/6J mice.MDSCs(expressing cell-surface CD11b and GR-1 antigens )from bone marrow were harvested 3 and 7 days after CLP and were separated with magnetic bead sorting technique .Then,cytokines secretion and arginase-I activity were detected and the T cell proliferation in vitro and the expression of miR-146a of MDSCs (3 d and 7 d after CLP)were observed.Results MDSCs secreted mostly such promoting inflammatory factors as TNF-α, IL-6 3 days after CLP, but 7 days after CLP , they primarily secreted IL-10 and TGF-βwhich were anti-inflammatory factors . MDSCs had potent immunosuppressive properties by increasing T cell suppression in a late anti-inflammatory phase ( CLP3 d vs CLP7 d, P<0.05).In the meantime,miR-146a of the MDSCs in bone marrow was overexpressed in septic mice at 7 days(P<0.05). Moreover,the expression of miR-146a of the MDSCs in bone marrow of septic mice was higher at 7 days than at 3 days after CLP(P<0.05).Conclusion The data indicate that the phenotype of MDSCs evolves through early pro -inflammatory phase into the late anti-inflammatory phase .MDSCs have potent immunosuppressive properties in the late phase of sepsis . miR-146 a might play a crucial role in the regulation of immunosuppressive activity of MDSCs in late sepsis .

ObjectiveTo investigate changes of Th17/Treg ratio in peripheral blood of patients with traumatic sepsis and discuss the underlying clinical significance.MethodsThrity-five multi-trauma patients enrolled in the study were divided into trauma group (n =22) and sepsis group (n =13 ).Another nine healthy volunteers were designed as the control group.Peripheral blood was collected from all the groups so as to determine the ratios of Th17 and Treg by flow cytometry,detect the mRNA expressions of RORγt and FoxP3 by real time fluorescence quantitative PCR and measure serum levels of IL-4,IL-6,IL-10,IL-17A,IL-23,transforming growth factor-β (TGF-β),interferon-γ (IFN-γ) and C-reaction protein (CRP) by ELISA.Further,the change of Th17/Treg ratio and its clinical significance were analyzed.ResultsThe proportion of Th17 in the trauma group showed no significant difference with that in the control group, whereas the ratio of Th17/Treg was lower than that in the control group (P ＜ 0.05).The proportion of Th17 and ratio of Th17/Treg in the sepsis group were higher than those in the trauma group and control group (P ＜ 0.01 ).Serum concentrations of TGF-β,IL-6 and IL-23 in the sepsis group were all significantly higher than those in the control group ( P ＜ 0.01 ).Serum levels of TGF-β had no significant difference between the sepsis group and the trauma group,while serum concentrations of IL-6 and IL-23 in the sepsis group were higher than those in the trauma group ( P ＜ 0.01 ).Serum level of IL-17A and mRNA expression of RORγt in the sepsis group were higher than those in the trauma and control groups ( both P ＜ 0.01 ).ConclusionTh17/Treg imbalance is closely correlated with the development of sepsis in patients with severe trauma.

In order to investigate the correlation between traumatic servity and blood cAMP and cGMP levels in the patients with acute trauma and its clinical significance, 120 cases of trauma were randomly selected and divided into 4 groups (n = 30 in each group): mildly traumatic group (ISS < or = 9), moderately traumatic group (ISS = 10-16), severely traumatic group (ISS = 17-25) and dangerously traumatic group (> 25). The cAMP and cGMP levels were assayed in sera, leucocytes and platelets respectively in 6 h and 24 h after trauma. The results showed that cAMP and cGMP levels were elevated significantly in sera and platelets (P < 0.05 or P < 0.01), meanwhile cGMP levels in leucocytes (P < 0.05). It was concluded that cAMP and cGMP might play an important role in traumatic stress, participate in the cellular signal transducation and promote the immune function of leucocytes and the coagulation founction of platelets. Serum cAMP and cGMP levels were upregulated correspondingly as ISS increased, and positively correlated to the traumatic severity.
Blood Platelets/metabolism ; Cyclic AMP/*blood ; Cyclic GMP/*blood ; *Injury Severity Score ; Leukocytes/metabolism ; Wounds and Injuries/*blood

Objective To investigate the expression and significance of human TREM-1 mRNA in patients with (biliary) infection. Methods Peripheral blood of 32 patients with biliary infection and 7 healthy volunteers were (collected). TREM-1 mRNA was determined by semi-quantitative RT-PCR. TNF-? was determined by ELISA method. Results The values of TREM-1/?-actin of control group was 0.48?0.072, while those of biliary infection group in 1d, 2d, 3d, 7d were 0.93?0.070,0.90?0.060,0.82?0.092,0.66?0.062 respectively (P

Objective To observe the effect of neuromuscular electrical stimulation (NMES) at different intensities on functional swallowing and the velocity of hyolaryngeal excursion in post-stroke dysphagia.Methods Thirty stroke survivors with dysphagia were randomly divided into a control group,an NMES group and an intensive NMES group,each of 10.All 3 groups received conventional swallowing training,while the patients in the NMES group also received NMES at 30 ～ 80 Hz in an intensity of ≤ 25 mA once a day and those in the intensive NMES group received it twice a day.Before,as well as 2 and 4 weeks after the treatment,video fluoroscopy when swallowing pap was used to determine the superior and anterior excursion distances of the hyoid and larynx and the excursion's duration and the corresponding velocity.A water drinking test (WDT) was also administered,and dysphagia severity scale (DSS) and penetration-aspiration scale (PAS) ratings were assessed.Results After the treatment,significant improvement was observed in the average WDT,DSS and PAS scores of all three groups compared to before the treatment.The average WDT score of the intensive NMES group was significantly better than that of the control group after 2 weeks of treatment.After 2 and 4 weeks of treatment,the average DSS of the intensive NMES group was significantly better than that of the control group,while the intensive NMES group's average PAS score was significantly better than those of both of the other groups.The average anterior hyoid excursion velocity of the intensive NMES group was significantly faster than those of the other two groups after both 2 and 4 weeks of treatment.Pearson correlation analysis indicated that the PAS score was significantly correlated with the anterior hyoid excursion velocity.Conclusion Two NMES sessions a day are superior to only one session in improving functional swallowing after stroke.It better promotes quick movement of the body parts involved.

Human monocyte leukemia cell line THP-1 was stimulated with lipopolysaccharide (LPS) to simulate the sepsis model and the expression of human glucocorticoid receptor-alpha (GR-alpha) mRNA in montocytes with endotoxin tolerance was investigated. THP-1 cells were cultured in serum-free medium, randomly divided into groups A, B, C, D and E, and stimulated with 0, 10, 10, 100, 0 ng/mL LPS for 24 h followed with 100, 100, 10, 100, 0 ng/mL LPS for another 24 h respectively. The expression of GR-alpha mRNA was detected by semi-quantitative reverse transcriptional polymerase chain reaction. Tumor necrosis factor-alpha (TNF-alpha) was determined by enzyme linked immunosorbent assay (ELISA). The results showed that the A values of GR-alpha/beta-actin in groups A, B, C, D and E was 0.607 +/- 0.006, 0.368 +/- 0.005, 0.484 +/- 0.008, 0.509 +/- 0.004 and 0.564 +/- 0.014 respectively with the difference being significant among the groups (P < 0.05). The GR-alpha mRNA expression was negatively correlated with the TNF-alpha expression (P < 0.01). It was concluded that the down-regulation of the expression of GR-alpha mRNA in endotoxin tolerance THP-1 cells might play an important role in the development of endotoxin tolerance in THP-1 cells.
Drug Tolerance ; Endotoxins/*pharmacology ; Leukemia, Monocytic, Acute/*metabolism ; Leukemia, Monocytic, Acute/pathology ; Monocytes/*metabolism ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Receptors, Glucocorticoid/*biosynthesis ; Receptors, Glucocorticoid/genetics ; Tumor Cells, Cultured

Objective To investigate the efficacy of emergency vacuum sealing drainage (VSD) in the treatment of crush syndrome. Methods A total of 102 patients with crush injury were randomly screened out and divided into emergency VSD group (n = 50), delayed VSD group (n = 22) and control group (n = 30). The variances of indices of crush syndrome after VSD between three groups were analyzed. Results The vital sign, urine volume, laboratory examination results were improved and the incidence rate of crush syndrome, complications and mortality rate were significantly decreased in emergency VSD group than those in delayed VSD group and control group (P < 0.05). For patients with crush syndrome, the incidence of complications, duration of hospital stay and mortality rate were significantly decreased in emergency VSD group and delayed VSD group than those in control group (P < 0.05). Conclusion Emergency VSD can suck the exudate of the necrotic tissues and the wounds, maintain draining wounds and prevent infection and hence is helpful to prevent and treat the crush syndrome.

Objective To observe change of T regulatory cells (Tregs) and its relation with in-jury severity and sepsis following severe muhiple injury. Methods A total of 60 patients were em-ployed in the study and divided into severe group (30 patients) and critical group (30 patients) based on ISS scores and into sepsis group (22 patients) and non-sepsis group (38 patients) based on complication of sepsis. The proportion of Tregs in peripheral blood in different groups was detected by flow cytometry at days 1,3,5 and 8 after injury. Results The proportion of Tregs was significantly increased at day 5 postinjury, with statistical difference compared with that at day 3 postinjury (P < 0.01). The proportion of Tregs remained increasing at day 8 pestinjury (P < 0.05). At day 8 postinjury, the proportion of Tregs in critical group was significantly higher than that in severe group (P < 0.01). At the same time, the proportion of Tregs in sepsis group was significantly higher than that in non-sepsis group (P < 0.05). Spearman correlation analysis showed a positive correlation of Tregs proportion with ISS score (rs =0.654, P < 0.01). Conclusions Tregs play an important role in suppression of T cell-mediated im-munity after severe injury. The variation of Tregs can help evaluate prognosis and predict the risk of com-plicating sepsis in patients with severe multiple injury.

Objective To investigate the clinical therapeutic effect of integrative treatment model and damage control theory in treatment of severe multiple injury. Methods A retrospective study was done on 116 patients with severe multiple injuries who were divided into integrative treatment plus damage control group, integrative treatment group and conventional treatment group. The relationship of correlative factors with mortality rate among three groups was compared before operation, during operation and after operation. Results All the indices in integrative treatment plus damage control group were better than other two groups before operation, during operation and after operation, with lower mortality rate (P<0.05). Conclusions Early use of integrative treatment model and damage control theory can obviously decrease complications, improve prognosis and decrease mortality rate for patients with severe multiple injury.

Objective The aim of this study was to investigate how different concentrations of dextran sulfate sodi-um ( DSS) influence the establishment of mouse model of inflammatory bowel disease ( IBD) and the effect of DSS on the expression of colitis-associated immune factors.Methods The DSS solution in different concentrations (3%, 5%, 7%) were given to male C57BL/6J mice to generate mouse inflammatory bowel disease model.The IBD mice were observed by defecation characteristics, body weight, and survival time.The animals were sacrificed at 6 days after the start of DSS drinking.The general appearance of colons was observed and scored.Moreover, the pathological changes of the colon were examined and analyzed by routine histology.The expression of immune factors in the spleen was detected by real-time PCR.Results The mice in the 3%, 5%, 7% DSS groups developed murine colitis.In addition, the incidence of IBD and mouse mortality rate was directly proportional to the increase of DSS concentration.Furthermore, the higher concentra-tion of DSS induced the expression of proinflammatory factors including TNF-α, IFN-γand IL-17A, but cause a decrease of anti-inflammatory factors such as IL-4, IL-10 and Treg-related transcription factor Foxp3.Conclusions Our data suggest that giving 5%DSS solution to C57BL/6J mouse is appropriate to efficiently establish a murine IBD model.This laid an important foundation for further studies of the pathogenesis of IBD, biological characteristics, and intervention factors.