连续性血液透析滤过对多器官功能障碍犬器官功能和细胞因子的影响

观察连续性血液透析滤过（CVVHDF）对多器官功能障碍综合征(MODS)犬器官功能和细胞因子的影响，并探讨CVVHDF治疗MODS的机制。 方法 15只Beagle犬采用失血性休克+复苏灌注+内毒素血症复制MODS模型，随机分为CVVHDF组(n=8)和MODS组(n=7)。内毒素注射完毕后CVVHDF组接受CVVHDF治疗12 h；MODS组未接受CVVHDF治疗。检测各时间点动脉血氧分压(PaO2)、动脉血二氧化碳分压(PaCO2)、谷丙转氨酶（ALT）、谷草转氨酶（AST）、Scr、BUN的水平。ELISA检测血浆TNF-α、IL-6、IL-10浓度。 结果 CVVHDF治疗后血浆IL-6、IL-10显著下降, 在3、6、9及12 h时间点显著低于MODS组，差异均有统计学意义（P < 0.01）；血浆TNF-α浓度无明显变化，两组间各时间点差异无统计学意义。超滤液中检出IL-6、TNF-α，筛选系数分别为0.27±0.13、0.1±0.1，但未能检出IL-10。CVVHDF组各主要器官功能均明显改善, 平均动脉压基本保持在正常水平，尤其在6、9及12 h时间点显著高于MODS组，差异均有统计学意义（P < 0.01）。PaO2逐渐升高，与MODS组在3、6、9及12 h时间点比较，差异均有统计学意义（P < 0.01）。 结论 连续性血液透析滤过能有效降低血浆IL-6、IL-10水平,使抗炎反应和促炎反应两方面趋于动态平衡；能明显改善内毒素诱导的低血压，提高动脉血氧分压。     

连续性静脉-静脉血液透析滤过对多器官功能障碍综合征犬不同溶质的清除作用

目的 观察连续性静脉-静脉血液透析滤过（CVVHDF）对多器官功能障碍综合征（MODS）犬病程中各种溶质的清除，为临床选择连续性肾脏替代治疗（CRRT）治疗方式提供理论依据。 方法 经失血性休克+复苏灌注+内毒素血症建立犬MODS模型, 随机分为MODS组（M组）和MODS+CVVHDF组（M+C组）。采用PRISMA机器行前稀释型CRRT，测定相同透析液及超滤液流量下一氧化氮（NO）、尿素氮（BUN）、血肌酐（Scr）、白细胞介素6 （IL-6）、IL-10、肿瘤坏死因子α（TNF-α）及内毒素（LP）浓度，计算其清除率。 结果 CVVHDF对小分子溶质如NO、BUN、Cr的清除率较高，对大分子物质IL-6、IL-10、TNF-α、LP的清除率较低。且随着各溶质分子量的增加，治疗时间的延长，各溶质的清除率呈现下降趋势。 结论 CVVHDF通过对流与吸附作用可有效清除MODS病程中的大、小分子溶质，是MODS救治中的强有力的支持治疗措施。     

连续性血液透析滤过对MODS犬肝肾IL-6和IL-10 mRNA表达的影响

目的：探讨连续性血液透析滤过（CVVHDF）后多器官功能障碍综合征（MODS）犬肝、肾组织白细胞介素-6（IL-6）和白细胞介素-10（IL-10）mRNA表达水平的变化及意义。方法：15只雄性Beagle犬,采用失血性休克＋复苏灌注＋内毒素血症复制MODS模型,随机分为CVVHDF组（n=8）和MODS组（n=7）,CVVHDF组在内毒素注射完毕后给予CVVHDF治疗12h,MODS组不给CVVHDF治疗。测定各器官功能相关指标,同时应用半定量逆转录-聚合酶链反应（RT-PCR）测定两组肝、肾组织中IL-6、IL-10mRNA表达水平。结果：CVVHDF组治疗后肝、肾功能有关指标水平均有不同程度的改善;与MODS组相比,在内毒素注射后3h及以后各时间点,血清肌酐（Scr）、尿素氮（BUN）水平显著降低（P〈0.05）;器官衰竭发生率较MODS组明显降低（37.5%vs85.7%,P〈0.05）;MODS组肝、肾组织IL-6mRNA表达水平显著高于正常对照组和CVVHDF组（P〈0.01）,而CVVHDF组肝、肾组织IL-10 mRNA表达水平显著高于正常对照组和MODS组（P〈0.01）。结论：连续性血液透析滤过能明显改善肾功能,CVVHDF早期应用可以降低MODS肝、肾组织IL-6/IL-10mRNA比值,有助于重建机体免疫系统内稳状态。     

The changes of atrial muscle RAS and ACE2-Ang(1-7)-Mas axis in MODS dogs treated by CVVHDF and the mechanism

Objective To observe the members’ dynamic change of renin-angiotensin system(RAS) and angiotensin converting enzyme 2(ACE2)-Ang(1-7)-Mas axis during the continuous veno-venous hemodiafiltration (CVVHDF) treating the dogs with multiple organ dysfunction syndrome (MODS), and to investigate the efficacy mechanism on cardiac function. Methods Dogs were subjected to hemorrhagic shock plus resuscitation and endotoxemia to establish MODS model, then they were randomly divided into 2 groups: CVVHDF group (n=8) and MODS group (n=6). After endotoxin injection completion, the CVVHDF group received CVVHDF for 12 h, MODS group didn't. Radioimmunoassay, euzymelinked mmunosorbent assay (ELISA) were used to detect the content of renin, AngⅠ, AngⅡ, Ang (1-7). Real-time PCR was used to detect the expression of renin mRNA and ACE2 mRNA. Western blotting was used to detect the protein content of renin, AngⅡ, ACE2 and Ang (1-7). Results (1) Organ function: Compared with the MODS group, the vital signs, heart, lung and renal function were significantly ameliorated in the CVVHDF group, the difference had statistical significance (P＜0.05). (2) RAS changes: To detect index from the level of organs, genetic and molecular in arterial tissue, the results displayed that the content of renin, AngⅠ, AngⅡ, the expression of renin mRNA, the protein content of renin, AngⅡ in CVVHDF group were lower than that in MODS group, the difference had statistical significance (P＜0.01). (3)ACE2-Ang(1-7)-Mas axis's changes: Using the same methods above to detect corresponding indicators, the results displayed that the content of ACE2, Ang(1-7), the expression of ACE2 mRNA and the protein content of ACE2, Ang(1-7) in CVVHDF group were significantly improved than that in MODS group, the difference had statistical significance (P＜0.01). Conclusions In the process of CVVHDF treating MODS, the ACE2-Ang(1-7)-Mas axis plays the role which antagonizes the RAS system, and to protect the cardiac function. This research may be important for MODS’ clinical rescue.     

The role of HMGB-1 on the development of necrosis during hepatic ischemia and hepatic ischemia/reperfusion injury in mice

BACKGROUND: High-mobility group 1 (HMGB-1) is a late mediator of endotoxin lethality in mice. The release of HMGB-1 is delayed compared to other proinflammatory cytokines that mediate shock and tissue injury. The purpose of this study was to investigate the role of HMGB-1 levels in response to hepatic ischemia, hepatic I/R injury, and the relationship between changes in HMGB-1 and other cytokines. MATERIALS AND METHODS: Three murine models were employed: our robust model of segmental hepatic warm ischemia (SHWI), a model of partial hepatic ischemia/reperfusion injury (PHIRI), and a model of total hepatic ischemia/reperfusion injury (THIRI). Over a 48-h period following ischemic insult and reperfusion using these models, serum HMGB-1 concentrations, concentrations of HMGB-1 in ischemic and nonischemic lobes, and serum concentrations of TNF-alpha and IL-6 levels were determined in mice. An anti-HMGB-1 antibody treatment was used in SHWI and THIRI to evaluate what aspects of response to ischemia and reperfusion were potentially mediated by HMGB-1. RESULTS: Hepatic HMGB-1 tissue concentrations exhibited biphasic changes in SHWI mice, which were increased in the ischemic lobes relative to nonischemic lobes at 12 h but decreased relative to nonischemic lobes at 24 h after ischemic insult. These results suggested that HMGB-1 was released into the systemic circulation by necrotic cells over the first 12 h but this process may be complete by 24 h postischemia. By 6 to 12 h after SHWI, serum TNF-alpha began to increase significantly and continued to increase for 18 h, followed by a sudden decline. Similarly, serum IL-6 increased over 1-3 h after SHWI and then decreased over the next 6 h. Treatment with an anti-HMGB-1 antibody significantly prolonged survival time in SHWI and THIRI. CONCLUSIONS: HMGB-1 plays a significant role in the response to hepatic ischemia and hepatic ischemia/reperfusion injury. The present study demonstrated a time-dependent production of HMGB-1 following hepatic warm ischemia in mice. The inherent HMGB-1 in ischemic areas was exhausted and HMGB-1 may be released by necrotic cells. HMGB-1 activation is involved in immediate proinflammatory stress response to I/R and anti-HMGB-1 antibody treatment remarkably improved survival. We demonstrated that systemic HMGB-1 accumulation was measured at an earlier phase of the hepatic ischemia and ischemia/reperfusion injury model than LPS-induced endotoxemia.     

Pattern of release and relationship between HMGB-1 and IL-6 following blunt trauma

Background

High mobility group box-1 (HMGB-1), a recently identified inflammatory cytokine, is implicated in the pathogenesis of several inflammatory, infective and neoplastic processes. Patterns of expression following blunt trauma have not been adequately reported in the literature. This study aimed to quantify the serum concentrations of HMGB-1 following blunt trauma, and assess its relationship with the more established interleukin 6 (IL-6).

Patients and methods

20 patients with median injury severity score 17 (range 9–36) sustaining closed diaphyseal fractures of the femur treated by intramedullary nailing were included in the study. Serum concentrations of HMGB-1 and IL-6 were measured at several time points during their treatment.

Results

A strong correlation was observed between admission and day 1 post-op concentrations of IL-6 and both the injury severity score (ISS) and the requirement for intensive care unit treatment. Serum concentrations of HMGB-1 did not demonstrate such a correlation. Around day 3 when IL-6 concentrations begin to fall, serum HMGB-1 concentrations were observed to increase.

Conclusions

IL-6 concentration measured early after admission is again shown to be strongly associated with overall injury severity and requirement for intensive care unit treatment. In contrast, HMGB-1 appears to be a late inflammatory mediator with levels becoming elevated once serum concentrations of IL-6 begin to fall. However, we were unable to demonstrate any relationship with injury severity or requirement for ICU care at any stage. These preliminary findings may form the basis for future research in this area.     

Endogenous HMGB1 is required in endotoxin tolerance

Background

High-mobility group box 1 protein (HMGB1), a downstream inflammatory response modifier in sepsis and endotoxemia, alters endotoxin tolerance by affecting cellular hyporesponsiveness and tumor necrosis factor α and interleukin 1 production.

Objective

Endogenous HMGB1 signaling mechanisms during low-dose lipopolysaccharide (LPS)-induced endotoxin tolerance were investigated.

Methods

BALB/c mice were preconditioned with either 0.1 mL low-dose LPS (0.2 mg/kg) or phosphate-buffered saline (PBS) (control) followed by treatment with three consecutive injections of anti-HMGB1, IgY (an nonspecific antibody), or PBS, at 2, 12, and 22 h, respectively, Mice were then subjected to 0.1 mL high-dose LPS (10 mg/kg) or PBS at 24 h. Serum and hepatic tissue samples were obtained 1 or 3 h after final treatments. Signaling mechanisms were further investigated in the serum and hepatic tissues of mice preconditioned with 0.1 mL HMGB1 (1 mg/kg), low-dose LPS (0.2 mg/kg), or PBS for 1 h, and then high-dose LPS treatment for 3 h.

Results

The signaling mechanisms involved in low-dose LPS preconditioning required enhanced endogenous HMGB1 expression and secretion. Neutralizing endogenous HMGB1 with anti-HMGB1 antibodies following low-dose LPS preconditioning altered endotoxin tolerance by increasing serum tumor necrosis factor α, reducing hepatic interleukin-1R-associated kinase M expression, and partially restoring nuclear factor κB in vivo. The translocation from nucleus to cytoplasm of endogenous HMGB1 in RAW264.7 cells was also observed during low-dose LPS–induced endotoxin tolerance.

Conclusions

Increased interleukin-1R-associated kinase M and decreased nuclear factor κB activity in endotoxin tolerance is associated with endogenous HMGB1 expression after low-dose LPS preconditioning. These findings provide a basis for a better mechanistic understanding and the development of safer clinical therapeutics utilizing induced endotoxin tolerance.     

血必净注射液对脂多糖诱导大鼠腹膜间皮细胞肿瘤坏死因子α和高迁移率族蛋白1表达的影响

目的 观察血必净注射液对脂多糖（LPS）作用下大鼠腹膜间皮细胞（PMC）肿瘤坏死因子α（TNF-α）和高迁移率族蛋白1（HMGB-1）表达的影响。 方法 胰蛋白酶消化法用于PMC的原代培养和传代,经鉴定,第3代细胞用于实验。细胞随机分组：（1）正常对照组;（2）不同浓度LPS（1、10、100 mg/L）组;（3）不同时间LPS组：10 mg/L LPS 作用PMC 2、6、12、18、21、24、36 h;（4）血必净组：10 mg/L LPS预孵育2 h后,加入不同浓度血必净（2、10、20 g/L）再作用4及22 h。RT-PCR法检测HMGB-1 mRNA的表达。ELISA法检测细胞培养上清液中TNF-α和HMGB-1的蛋白量。 结果 （1）LPS呈剂量依赖和时间依赖性显著上调PMC HMGB-1 mRNA和蛋白表达,与对照组相比,差异有统计学意义（均P < 0.05）。（2） LPS刺激后,PMC TNF-α蛋白表达显著高于正常对照组,差异有统计学意义（P < 0.05）,高浓度组更为明显,且36 h内表达出现双峰。（3）血必净可抑制TNF-α和HMGB-1的表达,与10 mg/L LPS组相比,差异有统计学意义（P < 0.05）,随血必净浓度增加,抑制作用增强。 结论 HMGB-1作为晚期炎性因子参与了腹膜透析相关性腹膜炎的病理过程。血必净可明显下调TNF-α及HMGB-1的表达,减轻腹膜炎性反应损伤。     

甘氨酸抑制脂多糖介导的鼠枯否细胞激活的时相选择及机制探讨

目的探讨甘氨酸抑制脂多糖介导的鼠枯否细胞激活效应相关机制和甘氨酸的最佳用药时机。方法将40只BALB/c小鼠分为内毒素组、预防组、早期治疗组和后期治疗组,每组10只。分离培养枯否细胞后,内毒素组加入脂多糖(10mg/L),预防组、早期治疗组和后期治疗组分别在加入脂多糖前24h、加入后0和4h加入甘氨酸(1mmol/L),分别在加入脂多糖后0、1、2、6和12h,采用逆转录聚合酶链反应及蛋白印迹法测定枯否细胞的白细胞介素1受体相关激酶4(IRAK4)mRNA和蛋白表达水平,用酶联免疫吸附法检测枯否细胞的核因子κB(NFκB)活性和培养上清液的肿瘤坏死因子α(TNFα)含量。结果脂多糖刺激后,预防组的IRAK4mRNA和蛋白表达、NFκB活性的相对峰值分别为3.64±1.13、34.54±10.31、0.47±0.10,TNFα峰值为(1780.70±210.17)pg/ml,与早期治疗组比较,差异均无统计学意义,但与内毒素组和后期治疗组比较,各峰值均明显降低,差异有统计学意义。结论提前或者在脂多糖刺激的同时应用甘氨酸,能有效抑制脂多糖介导的枯否细胞激活效应,其作用机制之一可能为抑制IRAK4的表达。     

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??Effect of continuous veno-venous hemofiltration on the plasma levels of endotoxin and cytokine in patients with severe intra-abdominal infection YANG Fan, ZHAO Yun-zhao, CHEN Yu, et al. Research Institute of General Surgery, Clinical School of Medical College, Nanjing University; Nanjing General Hospital of Nanjing Military Command, PLA, Nanjing 210002 China
Corresponding author ??ZHAO Yun-zhao, E-mail??yzzhaomd@gmail.com
Abstract Objective To investigate the clearance effect of continuous veno-venous hemofiltration??CVVH??on plasma endotoxin and cytokine in patients with severe intra-abdominal infection, and evaluate its clear ability and influence factors on endotoxin and cytokine in patients with severe intra-abdominal infection. Methods Twenty-one patients with severe intra-abdominal infection in ICU were enrolled for the CVVH treatment carried out between September 2010 and September 2011 in Nanjing General Hospital of Nanjing Military Command to observe the plasma changes of endotoxin, TNF-α, IL-10 during the treatment in 72h. The patients were divided into two groups: the filters were replaced every 24h group (n=10) and the control group (n=11). Results Plasma endotoxin and TNF-αdecreased significantly at 24h after CVVH treatment (P??0.05). Yet, the level of IL-10 began to decrease at 48h after CVVH treatment. The plasma endotoxin maintained on low level at 48h and 72h after CVVH treatment in the observation group compared with the control group(P??0.05). Conclusion Plasma endotoxin and cytokines (TNF-α, IL-10) can be removed effectively with CVVH in patients with severe intra-abdominal infection. CVVH has a positive effect on accelerate the removal of the plasma endotoxin and maintain a low level when replace the filter every 24 hours.     

Effect of CO2 pneumoperitoneum on the systemic and peritoneal cytokine response in a LPS-induced sepsis model

We studied the effect of carbon dioxide (CO2) pneumoperitoneum on the systemic and peritoneal cytokine response in a rat model of intraperitoneal sepsis. After intraperitoneal injection of bacterial lipopolysaccharide (LPS, 10 mg/kg), rats were divided into 3 groups (n = 49 in each group): control (abdominal puncture); CO2 pneumoperitoneum, and laparotomy. Blood and peritoneal lavage fluid (PLF) were sampled at 0, 1, 2, 3, 4, 6, and 8 h after LPS challenge. Blood cell counts, plasma endotoxin level, and the levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6) in the plasma and PLF were measured. Blood cell counts did not differ between the 3 groups. Plasma endotoxin levels in the pneumoperitoneum group were significantly increased immediately after the procedure (p < 0.05). Although peak plasma TNF-alpha levels in the pneumoperitoneum group were seen immediately after the procedure, other changes in plasma cytokine levels did not differ significantly between the 3 groups. PLF TNF-alpha and IL-1beta levels in the pneumoperitoneum group were significantly lower than levels in the control and laparotomy groups soon after the procedure (p < 0.05). PLF IL-6 levels in the pneumoperitoneum group tended to be lower than those in the laparotomy group. In conclusion, CO2 pneumoperitoneum might induce different responses between systemic and peritoneal cytokines soon after the procedure in a rat model of intraperitoneal sepsis.     

Differential expression of the immunoinflammatory response in trauma patients: burn vs. non-burn

Rationale

Cytokines are central mediators of the immune-inflammatory response to injury and subsequent multiple organ dysfunction syndrome (MODS). Although previous studies evaluated cytokine levels after trauma, differences between patients with burn and non-burn trauma have not been assessed systematically.

Methods

A prospective database of trauma patients admitted between May 2004 and September 2007 to the burn or surgical intensive care units within 24 h of injury with an anticipated stay of at least 72 h was analyzed. Sequential clinical and laboratory parameters were collected in the first week, including multiplex analysis data for plasma levels of inflammatory cytokines (IL-6, and IL-8). Patients with known pre-injury coagulopathy were excluded. A Marshall score of 10 or greater was defined as MODS.

Results

A total of 179 patients were enrolled (67 burn and 112 non-burn). Plasma IL-6 and IL-8 levels were markedly elevated in both burn and non-burn patients compared to healthy volunteers. Burn subjects had higher levels of IL-6 and IL-8 than the non-burn on days 1 through 7 after injury. Subjects with burns and at least 30% total body surface area were older and had a lower injury severity score, a higher prevalence of MODS, and correspondingly higher mortality. Multivariate analysis of injury type, MODS, and time did not demonstrate an influence of MODS.

Conclusions

Burns were associated with a greater and more sustained immune-inflammatory response than non-burn trauma as evidenced by elevated plasma IL-6 and IL-8 levels during the first week. There was no association between MODS and plasma cytokine levels.     

Circulating concentrations of adiponectin, an endogenous lipopolysaccharide neutralizing protein, decrease in rats with polymicrobial sepsis

BACKGROUND: Adiponectin is an anti-inflammatory cytokine that is specifically and abundantly produced by adipocytes as a secretory protein. A direct interaction between adiponectin and lipopolysaccharide (LPS) is not fully understood. To elucidate the effects of adiponectin on LPS, we first investigated interactions between recombinant adiponectin and LPS. MATERIALS AND METHODS: Various concentrations of LPS (50, 500, and 5000 pg/ml) and recombinant adiponectin (1, 10, and 100 microg/ml) were incubated for 1 h. The limulus amoebocyte lysate (LAL) activities in the mixture were measured. Interactions between adiponectin (100 microg/ml) and LPS (100 and 300 microg/ml) were also analyzed in Western blotting. Next, we determined plasma adiponectin, tumor necrosis factor-alpha (TNF-alpha), and endotoxin levels at 1.5, 3, and 24 h after onsets of rodent polymicrobial sepsis induced by cecal ligation and puncture (CLP). RESULTS: The incubation with adiponectin significantly and dose-dependently suppressed LAL activity in the mixture compared to control. Western blotting revealed that adiponectin incubated with LPS shifted to a higher mass. In the animal model of sepsis, both plasma endotoxin and TNF-alpha levels after CLP gradually increased and were significantly higher at 3, 24 h compared to those after sham operation. On the contrary, plasma adiponectin levels after CLP gradually decreased and were significantly lower at 3, 24 h compared to those after sham operation. Plasma adiponectin levels were negatively correlated with plasma endotoxin levels (r = -0.77, P < 0.01). CONCLUSIONS: Our results indicate that adiponectin might neutralize LPS in vitro and diminish LPS activity in rats with polymicrobial sepsis. These findings suggest that the anti-inflammatory effects of adiponectin are in part likely because of neutralization of LPS activity.     

In vivo expression of lipopolysaccharide binding protein and its gene induced by endotoxin

To investigate the expression of lipopolysaccharide binding protein (LBP) and its gene inrats with endotoxemia and explore the role of LBP in the response of host to endotoxin.     

Treatment of Acute Renal Failure in the Intensive Care Unit: Lower Costs by Intermittent Dialysis Than Continuous Venovenous Hemodiafiltration

Intermittent and continuous renal replacement therapies (RRTs) are available for the treatment of acute renal failure (ARF) in the intensive care unit (ICU). Although at present there are no adequately powered survival studies, available data suggest that both methods are equal with respect to patient outcome. Therefore, cost comparison between techniques is important for selecting the modality. Expenditures were prospectively assessed as a secondary end point during a controlled, randomized trial comparing intermittent hemodialysis (IHD) with continuous venovenous hemodiafiltration (CVVHDF). The outcome of the primary end points of this trial, that is, ICU and in-hospital mortality, has been previously published. One hundred twenty-five patients from a Swiss university hospital ICU were randomized either to CVVHDF or IHD. Out of these, 42 (CVVHDF) and 34 (IHD) were available for cost analysis. Patients' characteristics, delivered dialysis dose, duration of stay in the ICU or hospital, mortality rates, and recovery of renal function were not different between the two groups. Detailed 24-h time and material consumption protocols were available for 369 (CVVHDF) and 195 (IHD) treatment days. The mean daily duration of CVVHDF was 19.5 ± 3.2 h/day, resulting in total expenditures of €436 ± 21 (21% for human resources and 79% for technical devices). For IHD (mean 3.0 ±  0.4 h/treatment), the costs were lower (€268 ± 26), with a larger proportion for human resources (45%). Nursing time spent for CVVHDF was 113 ± 50 min, and 198 ± 63 min per IHD treatment. Total costs for RRT in ICU patients with ARF were lower when treated with IHD than with CVVHDF, and have to be taken into account for the selection of the method of RRT in ARF on the ICU.     

人高迁移率族蛋白B1、晚期糖基化终产物受体和C反应蛋白与脑外伤术后肺部感染严重程度的相关性及对预后的评估价值

目的探讨人高迁移率族蛋白B1（HMGB-1）、晚期糖基化终产物受体（RAGE）、C反应蛋白（CRP）与脑外伤术后肺部感染严重程度的相关性及对预后的评估价值。 方法前瞻性选取2016年5月至2018年5月于长治医学院附属和平医院接受治疗的182例脑外伤术后肺部感染者,根据肺部感染CURB-65评分系统标准将患者分为轻度组（74例）、中度组（60例）和重度组（48例）,并根据术后30 d临床结局分为生存组（151例）和死亡组（31例）。比较不同感染程度患者血清HMGB-1、RAGE、CRP水平并分析与肺部感染严重程度的相关性。通过Logistic回归分析明确影响患者死亡的危险因素,以受试者工作特征曲线（ROC）评价HMGB-1、RAGE和CRP对脑外伤术后肺部感染者死亡的预测价值。 结果重度组患者HMGB-1水平为（10.22 ± 2.35）μg/L,分别高于中、轻度组[（3.89 ± 1.01）μg/L和（2.00 ± 0.40）μg/L],差异有统计学意义（t = 18.821、29.502,P均< 0.001）;重度组患者RAGE水平为（9.01 ± 2.05）ng/L,分别高于中、轻度组患者[（5.89 ± 1.20）ng/L和（2.12 ± 0.22）ng/L],差异有统计学意义（t = 9.870、28.722,P均< 0.001）;重度组患者CRP水平为（50.33 ± 10.32）mg/L,分别高于中、轻度组患者[（32.33 ± 8.52）mg/L和（15.20 ± 3.52）mg/L],差异有统计学意义（t = 9.930、27.010,P均< 0.001）。中度组患者血清HMGB-1、RAGE和CRP水平均高于轻度组（t = 14.744、26.504、15.729,P均< 0.001）;血清HMGB-1、RAGE和CRP水平均与肺部感染CURB-65评分呈正相关（r = 0.696、0.763、0.851,P均< 0.001）。182例患者的病死率为17.03%（31/182）。死亡组患者呼吸机应用、气管切开、低蛋白血症占比均显著高于生存组,年龄、HMGB-1、RAGE、CRP、降钙素原（PCT）水平均高于生存组（P均< 0.05）。Logistic回归分析年龄、呼吸机应用、气管切开、低蛋白血症、HMGB-1、RAGE、CRP和PCT均为导致患者死亡的危险因素（OR = 2.016、2.423、2.252、1.853、2.606、2.199、1.919、1.904,P均< 0.05）。ROC曲线分析显示血清HMGB-1、RAGE和CRP预测脑外伤术后肺部感染者死亡的敏感性分别为78.02%、82.42%和85.16%,特异度分别为56.04%、69.78%和71.98%,曲线下面积（AUC）分别为0.756、0.801和0.882。 结论血清HMGB-1、RAGE和CRP水平与脑外伤术后肺部感染严重程度呈正相关,且均为导致患者死亡的危险因素,对于预测患者的死亡有较高的临床价值。     

Removal of inflammatory cytokines and endotoxin by veno-venous continuous renal replacement therapy for burned patients with sepsis

OBJECTIVE: To evaluate the effect of veno-venous continuous renal replacement therapy (CRRT) on the plasma levels of endotoxin and cytokines in severely burned patients with sepsis. METHODS: Twenty adult severely burned patients with sepsis were studied. For the diagnosis of sepsis, patients were randomly divided into CRRT (n=10) and Control (n=10). Both groups received conventional therapy after admission. Veno-venous CRRT was administered to 10 patients in the CRRT group whenever patients were determined to be septic. The plasma level of endotoxin, TNF-alpha, IL-1 beta, IL-6 and IL-8 were measured at 0, 1, 2, 6, 12, 36 and 60 h after CRRT initiation, and at 0, 12, 36 and 60 h after the patients were diagnosed as having sepsis in the Control group. MAIN RESULTS: Plasma level of endotoxin and all the cytokines after CRRT initiation were significantly lower than those before the treatment (P<0.01). The serial change of endotoxin, IL-1 beta, IL-6 and IL-8 was significantly lower at 12, 36 and 60 h after treatment compared with Control groups (P<0.01). A significant decrease in plasma TNF-alpha levels was seen at 36 and 60 h after treatment compared with Control groups (P<0.01). CONCLUSION: Plasma endotoxin and cytokines (TNF-alpha, IL-1 beta, IL-6 and IL-8) can be removed effectively with CRRT in severely burned patients with sepsis.     

Effect of high volume hemofiltration on CHOP protein from a canine model of multiple organ dysfunction syndrome

Objective To observe the effect of high volume hemofiltration (HVHF) on the expression of CCAAT enhancer binding protein(CHOP) during the treatment of multiple organ dysfunction syndrome (MODS). To investigate the role of CHOP protein act in apoptosis pathway mediated by the endoplasmic reticulum stress. Methods Twelve Beagle dogs were subjected to hemorrhagic shock plus resuscltation and endotixemia to establish MODS model, then they were randomly divided into two groups: HVHF group (n=6) and MODS group (n=6). After endotoxin injection completed, the HVHF group received HVHF treatment for 24 hours; MODS group did not receive. Vivo experiments: Blood samples were obtained at different time points(before operation, 0 h, 6 h, 12 h, 24 h after the injection of endotoxin). The dogs were killed and the tissue samples from lung, liver and kidney were took, then the expression of CHOP mRNA was determined. Vitro experiments: human umbilical vein endothelial cells (HUVECs) were induced by two groups’ blood samples to establish the apoptosis model. Gene expression, protein quantification and cell apoptosis rate were determined before and after the interference. Results Vivo experiments: The levels of CHOP mRNA from lung, liver and kidney had no significant difference between the two groups (P＞0.05). Vitro experiments: (1)The expression of CHOP mRNA: Compared with MODS group, the expression levels of CHOP mRNA were significantly decreased in HVHF group at 6 h, 24 h after the injection of endotoxin (P＜0.05). Compared with before, the expression levels of CHOP mRNA in the two groups were both significantly decreased after CHOP siRNA interference (P＜0.05). (2)The expression of CHOP protein: Compared with MODS group, the expression levels of CHOP protein were significantly decreased in HVHF group at each time points (P＜0.05). Compared with before, the expression levels of CHOP protein in the two groups were both significantly decreased after CHOP siRNA interference(P＜0.05). (3)Endothelial cell apoptosis rate: Compared with the preoperative rate, the two group’s endothelial cell apoptosis rate was decreased significantly at each time points(P＜0.05). Compared with MODS group, the endothelial cell apoptosis rate was significantly decreased in HVHF group at each time points(P＜0.05). Compared with before, the endothelial cell apoptosis rate in the two groups was both significantly decreased after CHOP siRNA interference(P＜0.05). Conclusion In the treatment of MODS process, HVHF can reduce endothelial cell apoptosis which may be related to the inhibition of CHOP mRNA expression and protein synthesis.     

Interleukin 1 and its relationship to endotoxin tolerance.

Endotoxin (lipopolysaccharide [LPS])-induced cytokine release has been implicated in the pathogenesis of sepsis. Sublethal doses of LPS induce tolerance to a septic insult. This study evaluated pretreatment with interleukin 1 (IL-1) against an LPS challenge and examined its relationship to endotoxin tolerance. C3H/HeN mice (N = 100) were injected intraperitoneally with phosphate-buffered saline (control group), IL-1 (200 micrograms/kg), or LPS (1 mg/kg) for 3 days. On day 5, peritoneal macrophages were harvested and assayed for antimicrobial activity (superoxide anion production and Candida albicans phagocytosis). Serum cytokine levels and survival after an LPS challenge on day 5 were also assessed. Pretreatment with IL-1 or LPS significantly increased superoxide anion production, C albicans phagocytosis, and survival compared with pretreatment with phosphate-buffered solution. Interleukin 6 levels significantly decreased in the IL-1 and LPS groups. Peak levels of tumor necrosis factor significantly decreased only in the LPS group. Thus, pretreatment with IL-1 or low doses of LPS may exert protective effects by decreasing levels of interleukin 6 while increasing antimicrobial activity. Mice pretreated with IL-1 were protected from endotoxin despite elevated peak levels of tumor necrosis factor, suggesting a different mechanism for endotoxin tolerance than for tolerance to tumor necrosis factor.     

丙酮酸乙酯对大鼠重症急性胰腺炎肾损伤的保护作用

目的 探讨丙酮酸乙酯对大鼠重症急性胰腺炎(SAP)肾损伤的治疗作用及其机制.方法 72只大鼠随机分为假手术组(S组),SAP组(P组)、丙酮酸乙酯治疗组(T组).大鼠SAP模型由5%牛磺胆酸钠胆胰管逆行注射诱发而成.各组于术后3、6、12 h检测血清中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、高迁移率族蛋白B...