脓毒血症并发急性肾损伤早期诊断标志物的研究

Objective To investigate the accuracy of NGAL in prediction of acute kidney injury (AKI) complicated by sepsis. Methods Blood and urine samples were collected at different time points in 74 sepsis patients, and 17 AKI patients were got from them.Cysteine Cys C levels were detected with Latex enhanced turbidimetric immunoassay (LETIA). Urine NGAL and Scr levels were detected with solid-phase sandwich enzyme-linked immunosorbent assay and creatinine enzymatic assay respectively. At the same time, 57 non-AKI patients were set as controls. Dynamic changes of urine NGAL and Scr were observed in sepsis patients in test and control group. ROC curve was used to evaluate the performance of AKI diagnosis by urinary NGAL.Results The Scr base line was (59. 38 ±16. 72) μmol/L.The median time of diagnose of AKI was 24 (12, 48) h past-sepsis diagnosis, while the Scr was(100. 35± 28. 26) μmol/L. Of the group, 17 cases of sepsis were diagnosed with AKI, accounting for 23% (17/74) prevalence. In sepsis group, base line value[(0. 61 ± 0. 15) mg/L] of serum Cys C was not upregulated at 2, 4 and 6 h, which was (0. 63 ± 0. 14) mg/L, (0. 68 ± 0. 16) mg/L and (0.65±0. 14) mg/L respectively. As to 8 h [(0. 85 ± 0. 22) mg/L], a slight upregulation was found with no significant difference (t = 1.63, P ＞ 0. 05).A trend of gradual increase of serum Cys C levels was found in sepsis AKI group at 12, 18, 24, 36, 48 and 72 h. A significant difference was found compared with base line (t = 2. 81, 2.98, 3.05, 3.11, 3. 38,3. 17,P ＜0.01). NGAL level[(96.21 ±45.32) μg/L] was significantly higher than base line value [(4. 98 ± 1.65) μg/L]. Subsequently, in each time point from the 2 h to 48 h, a trend of gradual increase in urinary NGAL levels was showed. The differences were significant compared with the baseline values (t =2. 74,2. 83,2. 91,3.04,3.15,3.22,3. 31,3.45,3.57 ,P ＜0. 01). Urinary NGAL levels of sepsis AKI group were significantly higher than those in non-AKI group at different time points with significant difference (t =2. 69,2. 73,2. 84,2. 96,3.02,3. 13,3.29,3.43,3.54,3.22,P ＜0. 01). ROC curve was selected to analyze diagnostic performance of urinary NGAL levels in AKI patients 2 h post-sepsis. Area under ROC curve was obtained as 0. 935, 95% confidence interval was 0. 683-0. 971. When cut-off value of urinary NGAL for patients 2 h post-sepsis was 50 μg/L, the sensitivity, specificity and accuracy were 94. 4%, 87. 5% and 96. 8% in AKI diagnosis respectively. Conclusions The occurrence of AKi can be accurately predicted by urinary NGAL levels 2 h post-sepsis , and its AKI diagnosis time is earlier than that of Scr. Urine NGAL can be used as early marker of AKI complicated by sepsis.     

脓毒症早期血清白细胞介素-18和10在疾病严重程度及预后评估中的价值研究

Objective To observe the dynamic changes in levels of serum interleukins (IL-18, IL-10)in the early stage of sepsis, and to appraise their values in the evaluation of severity and prognosis of sepsis.Methods Prospective randomized controlled study was conducted. Thirty-eight patients with sepsis who stayed longer than 72 hours in intensive care unit (ICU) from December 2009 to August 2010 were enrolled as sepsis group. At the same time, 20 patients without sepsis served as control group. The patients were classified as survival (n=12) or death group (n = 26) according to 28-day survival. The clinical laboratory examination data were recorded at 24, 48, 72 hours after admission to the ICU, and venous blood was obtained at the same time. The IL-18, IL-10 levels were determined by enzyme-linked immunosorbent assay (ELISA). Results The vital signs, blood routine, liver function, renal function, coagulation function,arterial blood gas, and electrolyte showed no significant difference between sepsis group and control group 24, 48, 72 hours after admission, the levels of IL-18 were lowered, IL-10 elevated, the IL-18/IL-10 ratio was lowered in the sepsis group, and all of them were higher than control group at each time point. The levels of IL-18, IL-10 in death group of patients with sepsis were all higher than those of survival group at 24, 48, and 72 hours [IL-18 (ng/L): 108. 36± 18. 54 vs. 91. 66±21. 49, 92.13±28. 92 vs. 54.16±31.76,91. 78 ± 17. 33 vs. 76. 04 ±22.09; IL-10 (ng/L): 99. 42 ± 12.10 vs. 77. 20 ±9. 47, 103. 39 ± 17. 24 vs.67.88±18.90, 118. 99 ±11. 20 vs. 99. 20± 12. 46, P＜0. 05 or P＜0. 01]. IL-18/IL-10 ratios were all lowered in both non-survivors and survivors with sepsis at 24, 48, 72 hours, while the differences were not statistically significant (1. 09±0. 19 vs. 1. 20±0. 32, 0. 92±0. 18 vs. 0. 98±0. 29, 0. 78±0.15 vs. 0. 77±0. 23, all P＞0. 05). Conclusion The levels of serum IL-18, IL-10 were all elevated in the early stage of patients with sepsis, and in non-survivors they were higher than those of survivors. With the progress of the illness, IL-18 showed a lowering tendency, while IL-10 showed an elevation. The levels of serum IL-18 and IL-10 may be valuable in evaluating the severity of sepsis and prognosis of patients with sepsis.     

脓毒症患者血清可溶性髓系细胞表达的触发受体-1水平及与预后的关系

Objective To approach the relationship between the contents of soluble form of triggering receptor expressed on myeloid cells-1 (sTREM-1) and prognosis in patients with sepsis. Methods Using prospective, control study design, a total of 50 patients with sepsis who were admitted in intensive care unit (ICU) of the Second Hospital of Tianjin Medical University from March to December in the year of 2009 were enrolled. Firstly, the patients were divided into sepsis (n=28) and severe sepsis (n=22) groups according to the patients' condition. Then the patients were divided into survival group (n = 34) and death group (n = 16)according to the clinical outcome at 28 days after onset of sepsis. Clinical and laboratory data including blood routine tests, blood chemistry, blood gas analysis, C-reactive protein (CRP) and procalcitonin (PCT) were collected on the 1st, 3rd and 7th day after onset. Acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ ) score was determined. sTREM-1 levels were determined using enzyme linked immunosorbent assay (ELISA) method. Correlation analysis of the sTREM-1, APACHE I score, white blood cell count (WBC) and CRP, using Logistic regression analysis. A total of 30 healthy persons were enrolled into the control group. Results The sTREM-1 levels (ng/L) in 50 septic patients on the 1st day were higher than those of the healthy persons (52. 80±9. 30 vs. 23. 29±6. 22, P＜0. 01). The sTREM-1 levels (ng/L) in severe sepsis group on the 1st, 3rd and 7th day (58. 25±10. 59, 65. 75±13. 57, 50.18±21. 73) were higher than those of the sepsis group (48.55±5.20, 42.85±8.54, 34.02±12.86, P＜0.05 or P＜0.01). The sTREM-1 levels (ng/L) of the survival group on the 1st, 3rd and 7th day (53. 07± 10. 47, 45. 04±9. 89,32. 84 ±8. 42) were decreased with the progression of the ailment. The sTREM-1 levels did not differ significantly between the control group and survival group on the 7th day (P＞0. 05). The sTREM-1 levels (ng/L) in the death group on the 1st, 3rd and 7th day were increased with the progression of the ailment (52.27±6.42, 69.67±12.83, 75. 70± 10. 55), and the level was significantly higher than that in survival group on the 3rd and 7th day (both P＜0. 01). The contents of sTREM-1 were positive correlated with APACHE Ⅱ score (r= 0.657, P＜0. 01), but not correlated with WBC (r= 0.023, P＞0. 05), whilesomewhat correlated with CRP (r=0. 150, P＜0.10). Logistic regression analysis showed that sTREM-1 [odds ratio (OR) = 0. 893,P = 0. 000] and APACHE I score (OR = 0.771, P = 0.000) might be potential prognostic factors for septic patients. The area under the receiver operator characteristic curve was 0. 868 and 0.930. The sensitivity of prognostic evaluation was 81.1% and specificity was 74.5% with sTREM-1 50 ng/L, and the sensitivity was 83. 8% and specificity was 86. 3% with APACHE Ⅱ score 20 to estimate the outcome. Conclusion The serum sTREM-1 are elevated at early stage in sepsis patients. It can reflect the severity of the condition. The sTREM-1 level, which might be considered as a potential prognostic factor for septic patients, is significantly correlated with APACHE Ⅱ score.     

脓毒症早期大鼠下丘脑-垂体-肾上腺轴超微结构变化与功能的关系

Objective To observe the changes in ultrastructure and function of hypothalamicpituitary-adrenal axis (HPAA), and to approach the relationship between them in early stage of sepsis in rats. Methods Thirty male Sprague-Dawley (SD) rats were randomly divided into normal control group,sham group, sepsis group. The sepsis model was reproduced by cecal ligation and puncture (CLP). The rats were sacrificed after collection of blood at 6 hours after CLP, and the levels of adrenocorticotropic hormone (ACTH) and corticosterone (CORT) in the plasma, and the corticotropin release hormone (CRH) in the tissue of hypothalamus were detected. The histopathological changes in HPAA were observed with transmission electron microscopy. Results The levels of ACTH and CORT in plasma, and the CRH in hypothalamus tissue of sepsis group were increased in the early stage of sepsis compared with the normal control group or sham group [ACTH (pmol/L): 5. 78 ± 0. 36 vs. 1.94 ±0.31, 2. 51 ± 0.10; CORT (nmol/L), 88.48±4.47 vs. 22.02±1.62, 34.20±2.51; CRH (μg/L): 101. 92±6. 61 vs. 61.65±6.05,66. 65±4. 03, P＜0. 05 or P＜0. 01]. The changes in ultrastructure of the hypothalamus, pituitary and adrenal were also found. In sepsis group, the ultrastructure of hypothalamus was as follows. Rough endoplasmic reticulum expansion and degranulation of rough endoplasmic reticulum, and swelling of Golgi complex were found. A large number of endocrine granules could be seen in ATCH cells in the pituitary with depletion of adrenal lipid droplets. Conclusion In septic rats, the HPAA was excessively activated, and ACTH and CORT in plasma, and CRH in hypothalamus were significantly increased in early stage of sepsis.The changes in ultrastructure of HPAA were obvious, and the change in function was closely related to the ultrastructural changes.     

伴有阻塞性睡眠呼吸暂停综合征的急性脑梗死患者血浆溶血磷脂含量的测定

Objective To observe the changing characteristics of plasma lysophosphatidic acid (LPA) or acidia phospholipid (AP) levels in patients with obstructive sleep apnea syndrome-associated(OSAS)acute cerebral infarction and to explore the pathophysiological mechanisms of OSAS-related stroke so as to provide basis for clinical antithrombotic therapy. Methods Thirty-six patients of OSAS, 32 patients of OSAS-related acute stoke and 36 patients of acute stoke without OSAS diagnosed by clinical and accessory examinations were enrolled in the current study. Thirty-eight age-matched healthy subjects were recruited as controls. The changes of the plasma LPA and AP levels were measured. Results Within 24 hours after symptom onset, the plasma LPA and AP levels in the OSAS-related acute cerebral infarction group (LPA(3. 78 ±0. 56) μmol/L; AP(7. 63 ± 1. 38) μmol/L) were significantly higher than those in the OSAS group(LPA(3. 17 ±0. 65) μmol/L; AP(6. 60 ± 1. 20) μmol/L) ,the not OSAS-related acute cerebral infarction group (LPA (3. 40 ± 0. 59)μmol/L; AP (6. 41 ± 1. 37)μmol/L) and the control group (LPA(2.76±0.45)μmol/L;AP(4.52±0. 83) μmol/L (P < 0. 01)) . The levels of LPA and AP in the OSAS group and the not OSAS-related acute cerebral infarction group were significantly higher than those in the control group(P<0. 01). Seven days after symptom onset, the plasma LPA and AP levels in the OSAS-associated acute cerebral infarction group (LPA(3.08 ± 0. 58) μmol/L; AP(6. 15 ±1. 14)μmol/L) were still higher(P < 0. 01) . The plasma LPA levels were not significantly different among the OSAS-related acute cerebral infarction group, the not OSAS-related acute cerebral infarction group and the control group 21 days after symptom onset, whereas the plasma AP levels in the OSAS-related acute cerebral infarction group (5. 04 ± 0. 83) μmol/L were still significantly higher than those in the not OSAS-related acute cerebral infarction group (4. 57 ± 0. 94) μmol/L and the control group (P < 0.05). Conclusions The significantly elevated plasma LPA and AP levels in patients with OSAS suggested that platelets in vivo are in an activated state and in cerebral ischemia and hypoxia state, especially for the OSAS-related acute cerebral infarction patients. The activated state of platelet may persist for a long time, thus the time window for antithrombotic therapy may be longer.     

伴有阻塞性睡眠呼吸暂停综合征的急性脑梗死患者血浆溶血磷脂含量的测定

Objective To observe the changing characteristics of plasma lysophosphatidic acid (LPA) or acidia phospholipid (AP) levels in patients with obstructive sleep apnea syndrome-associated(OSAS)acute cerebral infarction and to explore the pathophysiological mechanisms of OSAS-related stroke so as to provide basis for clinical antithrombotic therapy. Methods Thirty-six patients of OSAS, 32 patients of OSAS-related acute stoke and 36 patients of acute stoke without OSAS diagnosed by clinical and accessory examinations were enrolled in the current study. Thirty-eight age-matched healthy subjects were recruited as controls. The changes of the plasma LPA and AP levels were measured. Results Within 24 hours after symptom onset, the plasma LPA and AP levels in the OSAS-related acute cerebral infarction group (LPA(3. 78 ±0. 56) μmol/L; AP(7. 63 ± 1. 38) μmol/L) were significantly higher than those in the OSAS group(LPA(3. 17 ±0. 65) μmol/L; AP(6. 60 ± 1. 20) μmol/L) ,the not OSAS-related acute cerebral infarction group (LPA (3. 40 ± 0. 59)μmol/L; AP (6. 41 ± 1. 37)μmol/L) and the control group (LPA(2.76±0.45)μmol/L;AP(4.52±0. 83) μmol/L (P < 0. 01)) . The levels of LPA and AP in the OSAS group and the not OSAS-related acute cerebral infarction group were significantly higher than those in the control group(P<0. 01). Seven days after symptom onset, the plasma LPA and AP levels in the OSAS-associated acute cerebral infarction group (LPA(3.08 ± 0. 58) μmol/L; AP(6. 15 ±1. 14)μmol/L) were still higher(P < 0. 01) . The plasma LPA levels were not significantly different among the OSAS-related acute cerebral infarction group, the not OSAS-related acute cerebral infarction group and the control group 21 days after symptom onset, whereas the plasma AP levels in the OSAS-related acute cerebral infarction group (5. 04 ± 0. 83) μmol/L were still significantly higher than those in the not OSAS-related acute cerebral infarction group (4. 57 ± 0. 94) μmol/L and the control group (P < 0.05). Conclusions The significantly elevated plasma LPA and AP levels in patients with OSAS suggested that platelets in vivo are in an activated state and in cerebral ischemia and hypoxia state, especially for the OSAS-related acute cerebral infarction patients. The activated state of platelet may persist for a long time, thus the time window for antithrombotic therapy may be longer.     

脓毒症大鼠肺基质金属蛋白酶-2/9表达与血必净干预

Objective To detect the expression of MMP-2/9 and TIMP-1/2 in the lung of Vibrio vulnificus sepsis rats and observe the intervention of Xuebijing injection. Method One hundred and ten SD rats of clean grade were randomly(random number) divided into normal control group (group A, n = 10),Vibrio vulnificus sepsis group (group B, n = 50. Sepsis was reproduced in rats with subcutaneous injection in left lower limb with Vibrio vulnificus) and Xuebijin intervention group ( group C, n = 50. Rats were intraperitoneal(ip) with the dose of Xuebijing 4mL/kg at the time of 30 min later after infection). The rats in group B and C were sacrificed at 1 h, 6 h, 12 h, 24 h, 48 h after infection, the expression of MMP-2/9 and TIMP-1/2 were examined by PCR, Immunohistochemistry or ELISA methods, the lung permeability were measured by Coomassie Brilliant Blue method. Experimental data used single factor analysis of variance, and between groups by LSD method for pairwise comparison,P ＜0.05 statistically significant difference. Results The lung permeability increased both in group B and C compared with group A,and in group B were relatively higher. The lung MMP-2/9, TIMP-1/2mRNA expression in groups B and C compared with in group A was markedly higher, and reached the peak at 6 h(0. 344 ± 0. 108 ),6 h ( 1. 230 ± 0.377 ), 12 h (0.523 ±0.098),12 h(0.280±0.070) (P＜0.05) in group B while at 12 h(0.256 ±0.074),6 h(0.968±0.225) ,12 h(0.746 ±0. 316) ,12 h(0.356 ±0.035) (P ＜0. 05) in group C; the MMP-2/9mRNA expression in group C decreased(P＜0. 05) compared with the group B while the TIMP-1/2mRNA expression increased(P＜0. 05). The lung MMP-2/9, TIMP-1/2 protein expression in groups B and C compared with the group A(0.345±0.109) also increased, and the peak was at 12 h (0. 692 ± 0. 191 ), 12 h (0. 061 ±0.017) ,24 h(1384.42 ±91) ,24 h(41.04 ±3.60)in group B while at 24 h(0. 217 ±0.065) ,12 h(0. 045± 0. 013 ) ,24 h ( 1617.22 ± 103 ) ,24 h (47.66 ± 3.58 )in group C, the MMP-2/9 protein expression in group C was lower than in group B(P＜0.05), the TIMP-1/2 protein expression in group C was similar to in group B early while marked increased(P＜0.05)later. Conclusions MMP/TIMP imbalance was one of the mechanisms of the lung injury in the rats with Vibrio vulnificus sepsis, Xuebijing could restore the balance of MMP/TIMP ratio.     

慢性血吸虫感染对脓毒症小鼠保护作用的研究

Objective To preliminarily study the protective effect of chronic schistosoma japonica (SJ)infestation against sepsis in mice and its mechanism. Methods BALB/c male mice were used, and the experiment was divided into three parts. Experiment 1: chronic SJ infestation model was reproduced by SJ cercaria inoculation through abdominal skin for 8 weeks. Twenty mice were randomly grouped into normal group (n=10) and SJ group (n=10). The levels of interleukins (IL-4, IL-10), tumor necrosis factor-α(TNF-α) and interferon-γ (IFN-γ) in serum were detected by enzyme linked immunosorbent assay (ELISA).Real-time polymerase chain reaction (PCR) was employed to detect the levels of IL-10 mRNA and TNF-αmRNA in abdominal macrophages. This experiment was meant to evaluate immune state in mice with chronic SJ infestation. Experiment 2: lipopolysaccharide (LPS) was intraperitoneally injected to reproduce sepsis model. Thirty mice were randomly grouped into LPS group (n=15) and SJ-LPS group (n=15). The levels of cytokines were determined by ELISA at 0, 24, 48 and 72 hours after LPS injection. This experiment was meant to detect the effect of chronic SJ infestation in mice during the septic process. Experiment 3 : two types of sepsis model were reproduced by cecal ligation and puncture (CLP) and LPS injection, respectively. The survival rate of mice with chronic SJ infestation in 72 hours in either type of sepsis was evaluated. Results Experiment 1, compared with normal group [IL-4 (56.32±8.66) ng/L, IL-10 (48.17±7.23) ng/L],chronic SJ infestation showed an increase in serum IL-4 [(151. 35 ± 12. 24) ng/L] and IL-10 [(133. 22 ±11. 09) ng/L, both P＜0. 05]. Chronic SJ infestation also resulted in an increase in IL-10 mRNA expression (SJ group 4. 46±1. 82, normal group 1. 52±0. 60) and inhibited TNF-α mRNA expression (SJ group 1. 61±0.93, normal group 2. 32±1.03) in abdominal macrophages (both P＜0. 05), indicating that macrophages could be differentiated into alternative activated macrophages. Experiments 2 and 3 showed that the levels of serum IL-4 and IL-10 were increased at 0 hour after LPS injection, and then gradually decreased in SJ-LPS group, but the levels were still higher than those in LPS group at 72 hours [IL-4 (ng/L): 92. 2±7. 6 vs.41.5±4. 5; IL-10 (ng/L): 92. 1±7. 8 vs. 35. 6±4. 0, both P＜0. 05]; the levels of TNF-α and IFN-γ were increased at 24 hours, and then decreased in SJ-LPS group, and the levels were lower than those in LPSgroup at 72 hours [TNF-α (ng/L): 82. 9±5. 6 vs. 91. 5±5. 2; IFN-γ (ng/L): 44.1±4. 8 vs. 52. 6±4. 0,both P＜0. 05]. Therefore, chronic SJ infestation could improve the survival rate of mice with sepsis induced by CLP or LPS (CLP: 80% vs. 20%, LPS: 70% vs. 30%, both P＜0.05). Conclusion Chronic SJ infestation could elevate anti-inflammatory factors in septic mice, thus ameliorating the survival rate, so it has protective effect on mice with sepsis.     

Changes of PPARγ protein expression and activity in peripheral blood lymphocytes of patients with sepsis

Objective To investigate the changes of PPARγ protein expression and activity of peripheral blood lymphocytes in patients with sepsis and its association with severity and prognosis of sepsis. Method Ac-cording to the guidelines to sepsis set by ACCP/SCCM consensus conference in 2003, 48 patients with sepsis ad-mitted in Emergency and Surgical ICU from December 2007 to March 2008 were enrolled in this perspective study. Sixteen healthy individuals were selected as controls. Patients with metastatic tumors, autoimmune disease, AIDS or under immunosuppressive therapy were excluded. This study was approved by the ethical committee of Zhong-shan Hospital, Fudan University. All patients were divided into mild and severe sepsis groups. Patients were also divided into survivor and non-survivor groups as per 28-day mortality. Peripheral blood lymphocytes were isolated by using Ficoll density gradient centrifugation. PPART protein expression was determined by using Westem Blot-ting. The activity of PPARγ was analyzed by using EMSA. Differences among groups were analyzed by using one-way ANOVA. Results The protein expression and activity of PPARγ were significantly increased in mild sepsis patients (0.56±0.12 and 4.13±0.22, respectively) compared with both healthy controls (0.39±0.07 and 2.42±0.17, respectively) and severe sepsis patients (0.30±0.07 and 1.63±0.12, respectively) (P < 0.05). However, the protein expression and activity of PPARγ were obviously decreased in severe sepsis patients compared with healthy individuals and mild spsis patients (P < 0.05). Survivors from sepsis had significantly higher protein expression and activity (0.54±0.11 and 3.59±0.34, respectively) than non-survivors (0.21±0.08 and 1.94 ±0.25, respectively) (P < 0.05). Conclusions These data suggest that the protein expression and activity of PPARγ in peripheral blood lymphocytes might be valuable biomarkers in assessing the severity and outcome of pa-tients with sepsis.     

严多发伤并发症的早期控制

Objective To analyze the efficacy of early management of eomplications in patients with multi-ple traumas by applying the damage control theory. Method All total of 116 patients admitted to Wuhan Tongji Hospital between Jan 2006 and Jan 2008 were included in this retrospective study and were classified according to treatment method. Overall, 55 cases underwent damage control operations and 61 cases underwent conventional management. The groups of patients were compared in terms of parameters such as length of operation time, time to resuscitate from shock, lethal triad of death (LTD), complications and treatment outcomes after operation. Results The length of operation time was (67.43±19.52) min, resuscitation time from shock was (6.77±3.16) h and LTD was (11.54±4.10) h in the damage control group, and (163.95±55.41) min, (22.51±11.65) h and (34.55±5.63) h, respectively, in the conventional group, with significant differences between the two groups (P< 0.01). The per capita complication and mortality rates were 227.27 % and 3.64 %, respectively, in the dam-age control group and 363.93 % and 9.84%, respectively, in the conventional group (P <0.05). Couclusions Here, we demonstrated that early management of patients with multiple traumas undergoing operations by apply-ing the damage control theory significantly decreased the operation time, and decreased time to resuscitation from shock and LTD. Furthermore, this approach significantly reduced the incidence of comphcations and the mortality rate of patients with severe multiple traumas.