Objective To investigate the effect of recombinant human growth hormone (rhGH) on intestinal bacteria and endotoxin translocation in experimental obstructive jaundice. MethodsObstructive jaundice rat models were made and divided into three groups: sham operation (SO) group, obstructive jaundice (OJ) group and obstructive with rhGH (OG) group. The number in each group was 20. The mice in rhGH group underwent subcutaneous injection each day of Saizen, with the dose of 0.75 u/kg, while SO group and OJ group received nitric sodium injection. All these maitained for 2 weeks, then the animals were killed and the endotoxin were determined by limulus test, and bacterial cultures of ascites, blood, mesenchymal lymph node, kidney, spleen and liver were made, and the height of villi and the thickness of intestinal walls were examined.ResultsThe value of endotoxin in OJ group was (0.77±0.03) u/ml, higher than that in OG group and SO group, while it was (0.40±0.02) u/ml and (0.33±0.03) u/ml (Plt;0.01). The bacteria translocation rate in OJ group was 58.8%, much higher than that in OG group, which was 10.0% (Plt;0.01). There was no difference between OG group and SO group (Pgt;0.05). Villi height in OJ group was (183.39±11.09) μm, and thickness was (255.62±16.58) μm. While in OG group was (237.52±13.65) μm, and (320.81±14.34) μm (Plt;0.01) respectively.Conclusion rhGH has significant effect on protecting the injuried mucosa barrier in obstructive jaundice, and can decrease endotoxemia and bacteria translocation.
Objective To observe the effect of glutamine （Gln） on intestinal permeability after surgery of children, also its influence on the plama level of interleukin-2（IL-2）, endotoxin and synthesize of protein through a random nutrition trial. Methods Twenty children suffered from congenital heart disease were divided into Gln group and control group with random number table, 10 cases in each group. They were all given isonitrogenous and isocaloric total paraenteral nutrition after 24 h postoperatively. In Gln group the Dipeptiven [-N （2）-L-alanyl-Lglutamine] was used with 2 ml/kg · 24h additionly. Before operation, 24h and 96 h after operation, intestinal permeability, serum level of endotoxin, IL-2, C-reaction protein, prealbumine were measured. Results Intestinal permeability increased in 24 h after cardiac surgery in two groups, while the concentration of endotoxin also increased, 96 h after surgery the intestinal permeability recovered, but the endotoxin level did not decrease in control group （P〈0. 01）. Conclusion Utilization of Gln can improve immune suppression, elevate the IL-2 level, decrease the endotoxin concentration, alleviate the infection, but has no effect on the protein synthesis after congenital cardiac operation of children.
PURPOSE:To investigate and changes of the retina and the chorid induced by lipopolysaccharide (LPS)in Lewis rats and to compare the results obtained on tissue wholemounts and sections. METHODS:Immunohistochemistry was carried out both on wholemounts of the retina and the chorid-sclera complex and on ocular sections from normal Lewis rats and those after LPS injection. RESULTS:It was shown on the tissue wholemounts that monocytes were attached to retinal blood vessels and emigrated into the choroid as early as 4hrs after LPS injection. Severe involvement of the retina and macrophages into whole area of these tissues.Furthermore increasing number of major histocompatibility complex classⅡ(MHC classⅡ)positive cells was observed in the choroid.The results on tissue sections revealed that the retina and the choroid were both involved as videnced by infiltration of these cells at some time points after LPS injection. CONCLUSION:Wholemount technique provides undoubtful evidences to show that the retina and choroid are primarily and severely involuted after LPS injection.The endotoxin induced uveitis is,for the first time,presumed to be model for human generalized uveitis. （Chin J Ocul Fundus Dis,1996,12:33-36）
【Abstract】Objective To investigate the preventive role of selective decontamination of the digestive tract (SDD) in gut-originated endotoxemia in acute necrotizing pancreatitis (ANP). Methods A lethal model of ANP was reproduced in Wistar rats by retrograde infusion of artificial bile into the main pancreatic duct. Normal control group (n=6), sham operation group (n=6), ANP group (n=14) and ANP+SDD (polymycin E, tobramycin and nystatin mixture) group (n=8) were randomly devided. Visceral pathologic changes, serum levels of TNFα and IL-1β, intestinal bacterial flora, plasma D(-)lactate and endotoxin contents, as well as the mortality were examined at 72h after operation in each group. Results Necrosis and inflammation of pancreas, with a remarkable elevation of serum TNFα and IL-1β and intestinal flora disturbance (with E.Coli content risen significantly) were seen in ANP rats. Simultaneously, ANP rats displayed elevated plasma concentration of D(-)lactate and endotoxin. In SDD group, enterobacteraceae and yeast were markedly depressed, while anaerobes were well preserved, with the value of B/E 〔Bifidobacterium/E.Coli, log10(CFU/CFU）〕 elevated in the ileac mucous membrane (1.73±1.23 vs -0.37±0.72 in ANP group,P＜0.01) and in the caecum content (∞ vs 0.88±0.77). In addition, depressed levels of D(-)lactate 〔（3.95±1.83） mg/L vs （8.05±3.05） mg/L in ANP group,P＜0.01〕, endotoxin 〔（0.227±0.084） EU/ml vs （0.423±0.155） EU/ml in ANP group, P＜0.01〕 and TNFα 〔（15.41±10.32） ng/L vs （46.79±24.31） ng/L in ANP group P＜0.01〕 in systemic or portal vein were observed in the SDD group. Moreover, SDD group displayed a declined 72h mortality(14.3% vs 58.8% in ANP group, P=0.005). Conclusion ANP is associated with gut barrier disorder and gut flora imbalance, which may exacerbate the process of gut-originated endotoxin translocation. By protecting gut flora and gut barrier against disorder, SDD attenuates ANPrelated endotoxemia and improves the outcome. SDD is advisable for the prophylaxis of gut-originated endotoxemia complicated from ANP.
【Abstract】ObjectiveTo investigate the influence of CO2 pneumoperitoneum on intestinal mucosa permeability in rats with liver cirrhosis. MethodsFifty rats were randomly divided into following groups: control group (n=5), cirrhosis group(n=5) and pneumoperitoneum group (n=40); the pneumoperitoneum group was further divided into 8 mm Hg group(n=20) and 13 mm Hg group (n=20). Four time points were chosen, including 0.5, 2, 6, and 12 hours after the end of pneumoperitoneum. After rat models with cirrhosis were established successfully, the abdominal cavity was insufflated with CO2 and maintained under the pressures of 8 mm Hg and 13 mm Hg respectively for two hours. The portal venous blood was collected and the levels of Dlactic acid and endotoxin were measured. ResultsThe levels of endotoxin and Dlactic acid in cirrhosis group were much higher than those of control group（P＜0.05）. The levels of serum endotoxin and Dlactic acid in pneumoperitoneum group were higher than those of cirrhosis group(Plt;0.05) regardless of pressure and time point. The endotoxin level in 13 mm Hg group was higher than that of 8 mm Hg group on different time points (F=5.466, P＜0.05), but there was no difference in Dlactic acid level between both of them（F=0.415,Pgt;0.05）.ConclusionThe intestinal mucosa permeability is increased in rats with liver cirrhosis. It can be further increased under CO2 pneumoperitoneum with certain pressure and time and in a pressuredependent manner. The permeability can decrease after removal of pneumoperitoneum.
To observe the change in plasma endotoxin and cytokine during the early period of intra-abdominal infection (IAI) complicated by multiple system organ dysfunction (MSOD) in animals. Twenty rabbits were randomly divided in to two groups. One group received the operation of cecal ligation plus puncture (CLP) inducing IAI complicated by MSOD, and another group received sham operation as a control. All animals were placed in metabolic cages and maintained with intravenous infusion for one week. Plasma levels of endotoxin and cytokine (TNF, IL-1, IL-6) were determined seperately at the beginning (0 hour) or 1, 2, 3, 4, 5, 6 and 24 hours after CLP. Blood bacteria cultures and pathological examination of several organs were made when the animal was dead or killed. Results: The levels of plasma endotoxin, TNF and IL-6 were found to be significantly increased at one or two hours after CLP, the incidence rate of bacteriemia was 80% and the pathological alterations in the abdomen and organs were remarkale, with an average survival time of 84.1±39.0 hours in CLP group. No change in plasma IL-1 level was found in the CLP group. Conclusion: The plasma levels of endotoxin and cytokine (TNF and IL-6) do increase in the early period of IAI complicated by MSOD, and the change in plasma IL-1 is not obvious.
ObjectiveTo observe systemic inflammatory response (SIR)of patients in different stages after the onset of aortic dissection (AD), and preliminarily explore a new staging system of AD based on SIR. MethodsFrom September 2011 to February 2012, 46 AD patients were admitted to the Department of Cardiovascular Surgery, West China Hospital of Sichuan University. There were 33 male and 13 female patients with their age ranging from 22 to 77 years (53.2±13.6 years). Blood samples were collected in 9 different periods after the onset of AD (0-12 hours (T1), 12-24 hours (T2), 24-48 hours (T3, 1-2 days), 48-96 hours (T4, 2-4 days), 96-168 hours (T5, 4-7 days), 168-336 hours (T6, 7-14 days), 336-720 hours (T7, 14-30 days), 720-1440 hours (T8, 30-60 days) and > 1 440 hours (T9, > days))to measure blood concentrations of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), C-reactive protein (CRP), endotoxin (ET), white blood cell (WBC)and neutrophils (Neut). SIR changes after the onset of AD were summarized. ResultsBlood concentrations of different inflammatory mediators were all significantly elevated within 14 days (T1-T6), significantly decreased in 14-60 days (T7-T8), and returned to normal range 60 days (T9)after the onset of AD. Peak levels of ET and TNF-α appeared in T1 with 263.42±29.98 pg/ml and 86.75±18.83 pg/ml respectively. Peak level of IL-6 appeared in T2 with 95.70±22.64 pg/ml. Peak level of CRP appeared in T5 with 123.74±54.78 mg/L. There was no obvious peak level of WBC or Neut. ConclusionDisease progression of AD can be divided into 3 stages including acute stage (within 14 days), subacute stage (14-60 days)and chronic stage ( > 60 days)based on the degree of SIR.
Objective To observe the histopathologic features and expression patterns of tumor necrosis factor-alpha; （TNF-alpha;）, interleukin-1beta;（IL-1beta;） and Escherichia coli lipopolysaccharide (LPS) in the rat vitreous with LPS inducedendophthalmitis. Methods Wistar rats were randomly divided into saline control group (SC,136 rats）,endophthalmitis group （EO, 168 rats）and blank control group （BC,12 rats）.EO group received an intravitreal injection of 5 mu;l LPS; SC group received 5 mu;l sterile saline and no intervention for BC group.Six,12,24,48, and 72 hours,5 and 7 days after injection, intraocular inflammation were observed and the eyes and vitreous were collected for histopathological examination and measurement of TNF-alpha;, IL-1beta; and LPS expression. Results Severe inflammatory responses in the eyes were observed in EO group between six and 72 hours after LPS injection，ocular inflammation subsided seven days after LPS injection. In the vitreous, a peak neutrophil count was observed at 24 hours （1224.64plusmn;132.2） cells/eye that rapidly declined at 72 hours （342.25plusmn;47.7） cells/eye. The levels of TNF-alpha; and IL-1beta; in EO group were peaked at 24 hours with （996.18plusmn;89.45） and（5556plusmn;1440）pg/ L, respectively；Persisted at 48 hours and began to decline rapidly thereafter. Seven days after LPS injection, levels of TNF-alpha; and IL-1beta; returned to baseline with （22.16plusmn;5.84）and （73.7plusmn;18.7） pg/L, respectively. LPS concentration in EO group decrease rapidly at 72 hours with （11.03plusmn;3.41） ng and disappear on days 7 with (0.22plusmn;0.08) ng after LPS injection.Conclusions Massive neutrophils infiltration, high levels expression of TNF-alpha; and IL-1beta; and spontaneous elimination of bacterial elements in vitreous cavity were major pathologic characteristics in this experimental model. The expression patterns of TNF-alpha;,IL-1beta; were in accord with LPS clearance process.
The purpose of this study was to determine the contribution of endotoxin (ET) in ocurrence and progression of acute pancreatitis (AP). The results indicated that correlation of ET changes with multiple organ damage in AP. The degree of ET elevation correlated well with the severty of AP. The level of plasma ET of severe AP patients was much higher than that of mild AP patients (Ｐ＜0.05). The chance of multiple organ damage got greater while the plasma ET level got higher. Moreover, the severety change of severe AP correlated with the change of plasma ET level. In other words, the ET level was reduced while the disease was recovering, elevated while it was becoming worse and maintained high level in dead cases. We think that plasma ET level can be used as a reference for differenciating mild AP with severe AP and a predictor for the prognosis of AP.
ObjectiveTo investigate the regulatory roles and changes of M3 receptor subtype in lipopolysaccharide (LPS)-preincubated rabbit pulmonary arteries, and assess the mechanism of altered vascular reactivity in septic shock. MethodsPulmonary arteries with intact endothelium were isolated from 26 male New ealand white rabbits weighing 2.0 to 2.5kg. he isolated pulmonary arteries were randomized into two grouops, including a normal group with normal saline and darifenacin adminstration, and an endotoxin group with LPS-preincubation and darifenacin adminstration.he response of arteries to phenylephrine (100μmol/L) and acetylcholine(ACH)(1μmol/L, 10μmol/L, 100μmol/L)were measured in normal and darifenacin-preincubated circumstances. ResultsThe percentages of ralaxation to ACH (1μmol/L, 10μmol/L, 100μmol/L) were (0.095±0.034)%, (0.150±0.036)%, and (0.445±0.090)% in the normal group, and (0.044±0.016)%, (0.093±0.029)%, (0.311±0.028)% in the endotoxin (LPS 4μg/mL, 4h) group. After pretreatment with M3 receptor antagonist darifenacin on different concentrations, the EC50 values responding to ACH (1μmol/L, 10μmol/L, 100μmol/L) were 1.483, 2.757, 2.958 in the normal group, and 6.015, 6.242, 6.411 in the endotoxin group. After pretreatment with M3 receptor antagonist darifenacin on different concentrations, the inherent activity of a value to ACH (1μmol/L, 10μmol/L, 100μmol/L) were 0.0146, 0.0323, 0.0825 in the normal group, and 0.0124, 0.0245, 0.0556 in the endotoxin group. ConclusionsLPS pre-incubation can reduce the relaxation response to ACH, and M3 receptor subtypes mediated this relaxation response. LPS also reduce the M3 receptor subtype intrinsic activity, which may be one of the mechanisms of decreased relaxation response to ACH in pulmonary arteris after LPS pretreatment, and also one of the mechanisms of pulmonary hypertension in septic shock.