OBJECTIVE To study the effect of insulin intensive treatment on hyper-proteolysis of skeletal muscle in scalded rabbit with sepsis. METHODS Male white rabbits were randomly divided into scald group (S group), scald sepsis group (SS group), scald with insulin treatment group (SI group), scald sepsis with insulin treatment group (SSI group), and control group (C group), with 6 animals in each group. The extent of scald injury was 30% total body surface area (TBSA) III degree in depth, and the model of scald sepsis was replicated by administration of endotoxin (2 mg/kg) into the peritoneal cavity immediately after scald injury. Insulin intensive treatment was given with injection of insulin to keep the plasma glucose level in physiologic range (4.4-6.1 mmol/L) for SI group and SSI group. The contents of 3-methylhistidine (3-MH) in extensor digitorum longus (EDL) muscles and urine were determined with high performance liquid chromatography. The expressions of ubiquitin mRNA in the EDL muscle were assessed with Northern blot analysis. RESULTS The contents of 3-MH in EDL muscles and urine of S group and SS group were increased significantly as compared with C group (all P<0.01), and that of SI group and SSI group were decreased markedly compared with S group and SS group (all P<0.01), respectively. The mRNA expression of ubiquitin of S group and SS group was enhanced significantly compared with C group (all P<0.01), and that of SI group and SSI group was down-regulated notably compared with S group and SS group (all P<0.01), respectively. CONCLUSION The activity of ubiquitin system, one of important protein degradation pathways, and the proteolysis in skeletal muscle are enhanced after scald injury, especially so sepsis sets in, and insulin intensive treatment can effectively alleviate hyper-proteolysis of skeletal muscle through inhibiting the activity of ubiquitin system at gene level.
Modulation of the key immune cell subsets by biomaterial has emerged as a potential target to promote tissue repair and regeneration. Based on calcium alginate (Alg) and glycol chitosan (GC), an injectable double-network (DN) hydrogel has been developed as a scaffold for cell delivery and cell cocultured system. Previous studies have documented the interaction between dendritic cells (DCs) and GC or Alg hydrogel, but the potential effect of DN hydrogel on activation of DCs still remains unclear. This research was conducted to explore the immunomodulatory influence and underlying mechanisms of GC/Alg DN hydrogel on DCs in vitro and in vivo. Stimulation of DCs with DN hydrogel obviously induced the maturation of DCs in vitro. In vivo, DN hydrogel did not have obvious influence on the maturation of splenic DCs on postimplantation days 3, 10, and 30. Mechanistically, we found that DN hydrogel induced the maturation of DCs via phosphorylation of phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin in vitro. It provides a novel understanding of the immunomodulatory property of DN hydrogel on DCs, which may serve as potential target for designing immune-mediated regenerative strategies.
The definition of sepsis was updated to sepsis-3 in February 2016. However, the performance of the previous and new definition of sepsis remains unclear in China. This was a retrospective multicenter study in six intensive care unit (ICUs) from five university-affiliated hospitals to compare the performance between sepsis-1 and sepsis-3 in China. From May 1, 2016 to June 1, 2016, 496 patients were enrolled consecutively. Data were extracted from the electronic clinical records. We evaluated the performance of sepsis-1 and sepsis-3 by measuring the area under the receiver operating characteristic curves (AUROC) to predict 28-day mortality rates. Of 496 enrolled patients, 186 (37.5%) were diagnosed with sepsis according to sepsis-1, while 175 (35.3%) fulfilled the criteria of sepsis-3. The AUROC of systemic inflammatory response syndrome (SIRS) is significantly smaller than that of sequential organ failure assessment (SOFA) (0.55 [95% confidence interval, 0.46–0.64] vs. 0.69 (95% confidence interval, 0.61–0.77], P = 0.008) to predict 28-day mortality rates of infected patients. Moreover, 5.9% infected patients (11 patients) were diagnosed as sepsis according to sepsis-1 but not to sepsis-3. The APACHE II, SOFA scores, and mortality rate of the 11 patients were significantly lower than of patients whose sepsis was defined by both the previous and new criteria (8.6±3.5 vs. 16.3±6.2, P = < 0.001; 1 (0–1) vs. 6 (4–8), P = <0.001; 0.0 vs. 33.1%, P = 0.019). In addition, the APACHE II, length of stay in ICU, and 28-day mortality rate of septic patients rose gradually corresponding with the raise in SOFA score (but not the SIRS score). Sepsis-3 performed better than sepsis-1 in the study samples in ICUs in China.
Mitochondria are dynamic organelles and maintain their structure through two opposing processes of fusion and fission. In mammals, there are many factors which influence the equilibrium between mitochondrial fusion and fission. It has been known that mitochondrial fusion is controlled by mitochondria-shaping proteins, including the large GTPase mitofusins (Mfn1/2) and optic atrophy protein 1 (OPA1), whereas the protein Fis1 and dynamin-related protein 1 (Drp1) are the key mediators of mitochondrial fission. Recent data revealed the relationship between mitochondrial fusion/fission and Ca2+ signaling. Ca2+ handling is controlled by the change of mitochondrial fission-fusion balance. On the other hand, intra- and near-mitochondrial Ca2+ signals can modify mitochondrial morphology and cellular distribution.
To investigate the potential effect of bifidobacterial supplement on intestinal mucosal and biological barrier following severe burns.Seventy Wistar rats were randomly divided into burn control group (BC, n=30), burn + treatment group (BT, n=30), and sham-burn group (NC, n=10). Animals in BT group were fed bifidobacterial preparation (5x10(12) CFU/L) after 30% total body surface area (TBSA) full-thickness thermal injury, 1.5 ml, twice daily. Samples were taken on postburn days 1,3,and 5. The incidence of bacteria/endotoxin translocation and the counts of bifidobacteria, E.coli, and fungi in ileo-cecum mucosa were determined with standard methods. Ileum mucosal injuries were evaluated light and electron microscopically by blinded examiners.(1) The incidence of bacterial translocation were 42% and 16% in BC and BT groups on postburn day 3 (P=0.004 0), 30% and 6% on day 5 (P=0.002 0), respectively. Plasma endotoxin levels were markedly higher in BC and BT groups than that in NC group at the early stage postburn, while there was a significant decrease in BT group compared with BC group on postburn day 1 (P=0.04). (2) The bifidobacteria counts in ileo-cecum mucosa were reduced by 10- to 60-fold after thermal injury, while there was a remarkable increase in animals fed with bifidobacteria. The E.coli counts in ileo -cecum mucosa were increased by 20- to 30-fold on postburn days 1 and 3, whereas those markedly reduced after 3-day bifidobacteria-supplemented formula, tending to normal range.(3)There were obviously ileum mucosal lesions on postburn days 1 and 3,the ileum lesion scores remained significant higher in B C group than that in NC group on postburn 5 day (P<0.05).However,the damage d ileum mucosa was markedly repaired after 3-day bifidobacteria-supplemented formula, and almost restored 5 days later.The intestinal mucosal and biological barrier appear to be markedly damaged after severe burns. The supplement of exogenous bifidobacteria can facilitate the improvement of mucosal as well as biological barrier function, thereby reducing the incidence of bacteria/endotoxin translocation secondary to major burns.
To investigate the effect of tumor necrosis factor-α-induced protein 8 like-2 (TIPE2) on apoptosis of CD4+ T lymphocytes in a murine model of severe burn injury.A total of 140 male mice were randomly allocated into 6 groups. Small RNA interference technique was used to construct a siTIPE2-overexpressing lentivirus, and severe burn injury models were established in the mice. CD4+ T cells were purified from spleen of the mice, and the expressions of TIPE2, Smad2/Smad3, P-Smad2/P-Smad3 and Bcl-2/Bimprotein in CD4+ Tregs were detected. The changes in mitochondrial membrane potential and cytochrome C in CD4+ T cells were detected, and the activities of caspase-3, caspase-8, and caspase-9 were analyzed.Down-regulation of TIPE2 promoted the apoptosis of CD4+ T lymphocytes in siTIPE2-burn group, in which the protein expressions of P-smad2/P-Smad3 decreased, Bcl-2 expression increased and Bim expression decreased significantly as compared with the other groups (P<0.01 or 0.05). The mitochondrial membrane potential and cytochrome C expression in CD4+ T cells were down-regulated in siTIPE2-burn group (P<0.05) with a lowered caspase-3 activity compared with TIPE2-burn group (P<0.01) and decreased caspase-8 and caspase-9 compared with the other groups (P<0.05). The apoptosis rate was the highest in TIPE2-burn group, whose Smad2/Smad3 was higher than that in the sham group (P<0.05) and the expression of P-smad2/P-Smad3 significantly increased compared with the other groups (P<0.05). In TIPE2-burn group, the mitochondrial membrane potential in CD4+ T cells was decreased (P<0.01), the expression of cytochrome C increased markedly (P<0.01), and the activities of caspase-3, caspase-8, and caspase-9 were all obviously higher than those in the other groups (P<0.05).As an important immunoregulatory molecule, TIPE2 can promote the apoptosis of CD4+T lymphocyte in mice with sever burn injury.
To investigate the effect of apoptosis of CD4+ CD25+ regulatory T cells (Tregs) on proliferation as well as secretory function of effector T cells (Teff) and potential influence of Xuebijing injection on them in septic rats.A sepsis model was reproduced by cecal ligation puncture (CLP), and Wistar rats were randomly divided into the control group (n = 8), sham-operated group (n = 8), CLP group (n = 8), and Xuebijing injection treatment group (n = 8). CD4+ CD25+ Tregs in each group were separated by immunomagnetic beads isolate system on day 3, the apoptosis rate, expression of forkhead/winged helix transcription factor p3 (Foxp3) and cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) on Tregs were analyzed by flow cytometry, and secretion levels of interleukin (IL)-10 from Tregs were measured by ELISA. Following co-culture of CD4+ CD25+ Treg with CD4+ CD25- T cells (1:1) for 68 hours, proliferative activity of Teff was determined by MTT, and IL-2/sIL-2R alpha levels were measured by ELISA.The apoptosis rate of Tregs in control group was 12.03% +/- 0.89%, which was not significantly different from sham-operated group 9.48% +/- 2.17%. The apoptosis rate of Tregs in CLP group 5.87% +/- 0.44% was lower than that in control group (P < 0.01), and treatment with Xuebijing injection markedly enhanced the apoptosis of Tregs 27.29% +/- 2.48%. Compared to CLP group, expression of Foxp3, CTLA-4, and the secretion of IL-10 of Treg were significantly lowered in Xuebijing injection group (all P < 0.01). The Teff proliferative activity in response to ConA, and IL-2 levels of Teff in CLP group were significantly suppressed compared with control group (P < 0.01), and secretion of sIL-2R alpha in the supernatants was much higher than that of the control group. In comparison to the CLP group, inhibitory rate of Teff proliferative activity and the sIL-2R alpha levels were significantly decreased, while the secretion of IL-2 was increased in Xuebijing injection group (P <0.01).CD4+ CD25+ Tregs could markedly upregulate the suppressive function on Teff in sepsis, and treatment with Xuebijing injection effectively enhanced apoptosis of Tregs, thereby down-regulating the suppression on Teff.
The objective of this study was to evaluate the clinical efficacy of thiamine and vitamin C with or without hydrocortisone coadministration on the treatment of sepsis and septic shock.MEDLINE, EMBASE and CENTRAL databases were searched for randomized controlled trials (RCTs) that made a comparative study between the combination therapy of vitamin C and thiamine with or without hydrocortisone and the administration of placebo in patients with sepsis or septic shock. Two reviewers independently performed study selection, data extraction and quality assessment. Both short-term mortality and change in the sequential organ failure assessment (SOFA) score from baseline (delta SOFA) were set as the primary outcomes. Secondary endpoints included intensive care unit (ICU) mortality, new onset of acute kidney injury, total adverse events, ICU and hospital length of stay, duration of vasopressor usage and ventilator-free days. Meanwhile, trial sequential analysis was conducted for primary outcomes.Eight RCTs with 1428 patients were included in the current study. The results showed no significant reduction of short-term mortality in sepsis and septic shock patients who received combination therapy of vitamin C and thiamine with or without hydrocortisone compared to those with placebo {risk ratio (RR), 1.02 [95% confidence interval (CI), 0.87 to 1.20], p = 0.81, I2 = 0%; risk difference (RD), 0 [95% CI, -0.04 to 0.05]}. Nevertheless, the combination therapy was associated with significant reduction in SOFA score [mean difference (MD), -0.63, (95% CI, -0.96 to -0.29, p < 0.001, I2 = 0%] and vasopressors duration (MD, -22.11 [95% CI, -30.46 to -13.77], p < 0.001, I2 = 6%). Additionally, there were no statistical differences in the pooled estimate for other outcomes.In the current meta-analysis, the combination therapy of vitamin C and thiamine, with or without hydrocortisone had no impact on short-term mortality when compared with placebo, but was associated with significant reduction in SOFA score among patients with sepsis and septic shock.
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