The serum activities of 3 cytokines (TNF,IL-1 and IL-6) were observed in 23 patients admitted within 4 days of onset of acute pancreatitis (AP). The results showed that the serum level of 3 cytokines raised in all of the AP patients, significant difference between TNF and IL-1 was abserved at admission and IL-6 did after one week of admission, suggesting that proper cytokine criteria are useful in predicting severity of the disease but the relationship between cytokines and MOF had not established.
Objective To explore the possible anti-inflammatory mechanism of intensive insulin therapy （IIT） by studying the effect of IIT on the levels of TNF-α, IL-6, C-reactive protein （CRP） and APACHE Ⅱ score in biliary pyemia. Methods Twenty eight patients with biliary pyemia who were admitted by our department and given an operation within 24 h form Jan. 2005 to Dec. 2008 were randomly divided into two groups by using random number table numbers: one group treated with IIT （IIT group, n=14） and another group treated with routine insulin therapy （RIT group, n=14）. The inflammatory factors, such as TNF-α, IL-6 and CRP were detected dynamically and the APACHEⅡ score was calculated. ResultsThe level of CRP and APACHEⅡ score on day 5 and 7 and the levels of TNF-α and IL-6 on day 3, 5 and 7 after operation in IIT group were significantly lower than those in RIT group （P＜0.05, P＜0.0１）. Compared with preoperative levels, the IL-6 and APACHEⅡ score in IIT group commenced to decrease on day 3 after operation （P＜0.05）, that was earlier than control group. Conclusion The treatment with IIT can suppress the composition of TNF-α, IL-6 and CRP, protect impaired hepatic cells, and reduce APACHEⅡ score, the degree of systemic inflammation and incidence of MODS.
Objective To investigate the relationship between the level of serum-insulin like growth factor-1( IGF-1) and the nut ritional status of cancerous cachexia. Methods Colon cancer CT-26 cells were implanted subcutaneously to 30 liver2specified IGF-1 gene deleted (L ID) C57BL/ 6 mice to establish cancerous cachexia model and theother 30 C57BL/ 6 mice were included as cont rol group. The serum levels of IGF-1 , cytokine TNF-αand IL-6 , bloodglucose , albumin and t riglyceride were detected respectively on day 14 , 18 and 22 af ter the plantation of tumor. Thebody weight of mice , tumor weight and the weight af ter tumor removed in two group s were measured respectively.Results Af ter the plantation , the levels of IGF-1 in L ID group at different times were all significantly lower thanthose in cont rol group ( Plt; 0. 05) . The serum levels of TNF-α, IL-6 , blood glucose and t riglyceride were ascendinggradually over time ( Plt; 0. 05) , but weight s af ter tumor removed and the level of albumin were descending in twogroup s ( Plt; 0. 05) . Compared with the cont rol group , the serum levels of IL-6 , TNF-α, blood glucose and t riglyceride in L ID tumor-bearing mice were all significantly higher at different time point s ( P lt; 0. 05) . On day 18 and 22 ,the weight s af ter tumor removed and the amount of ingestion in L ID group were significantly lower than those in thecont rol group ( Plt; 0. 05) . Conclusion Compared with the low level of IGF-1 in cancerous cachexia , normal level ofserum IGF-1 may represent lower degree of cancerous cachexia2related cytokines and better nut ritional state , whichmay provide a novel idea of the therapy of cancerous cachexia.
Objective To observe the protective effects of ambroxol hydrochloride ( AMB) on rabbit model of acute lung injury ( ALI) induced by oleic acid and explore its mechanisms. Methods The ALI model of rabbit was induced by oleic acid. Twenty-four Japanese white rabbits were divided into three groups randomly, ie. a normal saline group ( NC group) , an ALI group and an ALI plus ambroxol injection group ( AMB group) . The pathological changes and apoptotic index ( AI) in lung tissue, Caspase-3 activity in lung tissue homogenate were observed 6 hours after the intervention. Serum activity of superoxide dismutase ( SOD) and serum levels of malonaldehyde ( MDA) , interleukin-1β( IL-1β) , and tumor necrosis factor-α ( TNF-α) were measured simutanously. Results The pathological injury of lung in the AMB group was milder than that in the ALI group. Both the AI in lung tissue and Caspase-3 activity in homogenate in the AMB group were lower than those in the ALI group significantly ( P lt;0. 01, P lt;0. 05 respectively) , butwere higher than those in the NC group( both P lt; 0. 01) . The activity of SOD in serum measured 6 hours after AMB intervention was higher while the serum levels of MDA, IL-1βand TNF-αin serum were lower ( P lt;0. 01) than those in the ALI group significantly ( all P lt;0. 01) . Conclusions Ambroxol hydrochloride has protective effects on oleic acid-induced acute lung injury. The mechanisms may be related to inhibition of oxidative stress and suppression of cytokines synthesis ( IL-1βand TNF-α) , the activity of the Caspase-3,and the apoptosis of lung tissue.
The aim of this study was to investigate the role of tumor necrosis factor (TNF), interleukin-6(IL-6), C-reactive protein (CRP) in pancreatitis and its systemic complications. Thirty six patients with acute pancreatitis were studied, 12 with mild disease, and 24 severe disease, of whom 9 developed systemic complications. TNF, IL-6, CRP in these patients with pancreatitis was assessed during the first, 4th, 8th days of admission. The serum concentration of TNF, IL-6, CRP were significantly increased, and significantly higher in complicated group than in mild group and severe group. These findings suggest that proinflammatory cytokines play a central role in the pathophysiology of the disease, the host systemic response to pancreatic inflammation and the level of the response did relate to the development of organ dysfunction.
Objective To study the effect of bone marrow mesenchymal stem cells(MSCs) implantation into infarcted myocardium on cytokine secretion and angiogenesis. Methods 24 Guizhou xiang porcine were equally divided into experimental group and control group randomly. Three ml bone marrow was extracted from the posterior superior iliac spine. MSCs were cultured according to the methods of Wakitani’s. After being co-cultured with 5-azacytidine for 24 hours, these cells were labeled with bromodeoxyuridine(BrdU). Autologus MSCs were implanted into the acute myocardial infarct site both via the distal segment of the ligated left anterior descending artery (LAD) and topical injection. 3 amp; 6 weeks after transplantation, the samples from experimental group and control group were collected to detect the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and Ⅷ factor by immunohistology and video image digital analysis system. Results The expression of VEGF, bFGF and the microvessel counts in the experimental group were much higher than those of control group (Plt;0.01) at 3 and 6 weeks after transplantation. Conclusion MSCs, after being implanted into infarcted myocardium, shows the ability of secreting VEGF, bFGF, with subsequent angiogenesis.
【Abstract】Objective To observe concentration changes of substances including hepatocyte growth factor(HGF),interleukin-6(IL-6),nitric oxide(NO),nitric oxide synthase(NOS) that were secreted by sinusoidal endothelial cells(SECs) in different shear stress and study the effects of shear stress on the secretions of SECs.Methods To establish a hemodynamics equipment and to set up a way to separate and culture SEC. The rats were divided into two groups: the control group (shear stress was 0 dyn/cm2) and study group. Study group was divided into three subgroups according to shear stress (12, 24 and 48 dyn/cm2). Sinusoidal endothelial cell culture mediums were examined in different shear stress and phases.Results HGF, IL-6, NO and NOS secreted by sinusoidal endothelial cells in different shear stress were distinctive and had positive correlation with shear stress and action time. Conclusion In vitro, the secretion of HGF,IL-6,NO and NOS of sinusoidal endothelial cells are increased along with shear stress rising, which suggests that the rising pressure in portal vein after hepatectomy may result in SECs activated．Activated SECs then secret many cytokines and medias to trigger the regeneration of liver.
ObjectiveTo determine the nuclear factor kappa B (NFkB) activity in peripheral blood mononuclear cells (PBMC) in patients with acute cholangitis of severe type (ACST) and correlate the degree of NFkB activation with severity of biliary tract infection and clinical outcome.MethodsTwenty patients with ACST were divided into survivor group (14 cases) and nonsurvivor group (6 cases). Other 10 patients undergoing elective gastrectomy or inguinal hernia repair were selected as control group. Peripheral blood samples were taken 24 hours after operation, PBMC was separated and nuclear proteins were isolated from PBMC, and NFkB was determined with electrophoretic mobility shift assay (EMSA). The levels of TNFα, IL6 and IL10 in plasma were determined by using an enzymelinked immunoassay (ELISA). ResultsThe NFkB activity was 5.02±1.03, 2.98±0.51 and 1.02±0.34 respectively in three groups. It was increased in all patients with ACST, versus the control group (P＜0.05), and the patients of nonsurvivor group had higher levels of NFkB activation than those of survivor group (P＜0.05). The levels of TNFα and IL6 were (496.28±52.35) ng/L and (578.13±67.72) ng/L in nonsurvivor group; (284.47±39.41) ng/L and (318.67±34.92) ng/L in survivor group; (89.43±10.39) ng/L and (101.27±13.47) ng/L in control group. All patients with ACST had increased levels of TNFα and IL6, which were many fold greater than that of control group, and there was an evidence of significantly higher levels in nonsurvivor group than in survivor group (P＜0.05). All patients had also increased levels of IL10 as compared to control group (P＜0.05), but the IL10 concentrations in plasma were not significantly higher in nonsurvivors than that of in those survivors (Pgt;0.05). ConclusionNFkB activation in PBMCs in patients with ACST
Objective To investigate the effect of inducible nitric oxide synthase (iNOS) inhibitor aminoguanidine on pancreas islets cultured with cytokines TNF-α and IL-1β in rats. Methods Islets isolated from Wistar rats were purified and cultured. According to whether cytokines TNF-α, IL-1β and aminoguanidine were added into the medium respectively or not, islets were divided into 4 groups: cultured with islet only was taken as blank control group, cultured with TNF-α+IL-1β as cytokine group, cultured with aminoguanidine as aminoguanidine group, and cultured with TNF-α+IL-1β and aminoguanidine as aminoguanidine+cytokine group. NO level in culture medium and iNOS activity in islets tissue (Test Kit), apoptosis (TUNEL method) and viability of islets cell (acridine orange/ethidium bromide stain), and the function of islets (insulin release test) were measured. Results Compared with blank control group, the activity of iNOS in islet tissue and level of NO in culture medium increased, and the mass mortality and apoptosis appeared in islet cells, while insulin secretion decreased in cytokine group (P＜0.01). Compared with cytokine group, the activity of iNOS 〔（3.17±0.51） U/ml vs. （38.93±4.72） U/ml〕 and level of NO 〔（50.5±10.4） μmol/L vs. （313.0±35.4） μmol/L〕 decreased, the survival 〔（72.73±3.14）% vs. （57.07±5.07）%〕 increased and the apoptosis rate 〔（20.11±8.48）% vs. （41.17±6.87）%〕 decreased, the insulin secretion (secretion index: 3.50±0.27 vs. 1.96±0.19) improved； There were all significant differences in 2 groups (P＜0.01). Conclusion The iNOS inhibitor aminoguanidine could prevent the islet from the damage of iNOS/NO, alleviate the impairment of cytokines to islets, and ameliorate the survival and function of islets.
Objective To investigate the effects of recombinant adeno-associated virus type-2 (rAAV2) mediated delivery of pigment epitheliumderived factor (PEDF) on oxygen-induced retinal neovascularization (OIRNV) in mice. Methods A total of 22 C57/BL6 mice at the age of 3 days received intravitreal injections of 1 mu;l rAAV2-PEDF and rAAV2EGFP into the left eyes (experimental group) and the right eyes (control group). All mice were put into the oxygen box right after the injection to induce the OIRNV model.4 mice were sacrificed and PEDF protein in retina was measured by western blot at postnatal days 13 (P13). Twelve mice underwent retinal angiography with high molecular weight fluoresceindextran, and another 6 mice were sacrificed for retinal lectin immunohistochemistry staining at P17. Absolute and relative nonperfusion areas of retinal neovascularization were analyzed by Image-Pro Plus 5.1 software. Results The expression level of PEDF protein was higher in the experimental group than that in the control group.The absolute nonperfusion area was (0.96plusmn;0.22) mm2 in the experimental group and (1.96plusmn;0.34) mm2 in the control group; the difference between the two groups was significant（t=-8.554, P＜0.01）. The relative nonperfusion area was (8.64plusmn;1.52)% in the experimental group and (17.27plusmn;2.98)% in the control group with a significant difference between the two groups (t=-8.97, P＜0.01).The absolute area of retinal neovascularization was (0.37plusmn;0.11) mm2 in the experimental group which was obviously higher than (1.26plusmn;0.38) mm2 in the control group (t=-7.8, P＜0.01); the relative areas in experimental and control groups was (3.96plusmn;0.66)% and (11.45plusmn;2.06)%, respectively, whose difference is apparently(t=-8.51, P＜0.01).The areas of retina neovascularization were (0.11plusmn;0.003) mm2 and (0.41plusmn;0.02) mm2 in the experimental and control groups, respectively, and the difference between the two groups was significant（t=-5.14, P＜0.01）.Conclusions PEDF protein can stably express in the mice retina after rAAV2-PEDF transfetion. rAAV2-PEDF can decrease the retinal non-perfusion areas and inhibit the retinal neovascularization in OIRNV mice.