【Abstract】Objective To study the influence of early hemofiltration on plasma concentrations of proinflammatory cytokines TNF-α and IL-1β and their transcription levels in severe acute pancreatitis (SAP) pigs. Methods The model of SAP was induced by retrograde injection of artificial bile into pancreatic duct in pigs. Animals were divided randomly into two groups: SAP hemofiltration treatment group (HF group, n=8) and SAP no hemofiltration treatment group (NHF group, n=8). TNF-α and IL-1β plasma concentrations were measured by ELISA. Their transcription levels in the tissues of pancreas, liver and lung were assayed by semi-quantitative reverse transcription polymerase chain reaction. Results After hemofiltration treatment, the plasma concentrations of TNF-α and IL-1β increased gradually but were lower than those of NHF group at the same time spot 〔at 6 h after hemofiltration treatment, (618±276) pg/ml vs (1 375±334) pg/ml and (445±141) pg/ml vs (965±265) pg/ml, P＜0.01〕. At 6 h after hemofiltration treatment, the transcription levels of TNF-α and IL-1β in tissues of pancreas, liver and lung were lower than in NHF group (57.8±8.9 vs 85.7±17.4, 48.0±8.1 vs 78.1±10.2, 46.2±9.6 vs 82.4±10.5; 55.9±9.0 vs 82.2±15.7, 40.6±9.2 vs 60.0±10.6, 35.7±9.8 vs 58.1±9.3, P＜0.01). Conclusion Early hemofiltration can reduce TNF-α and IL-1β plasma concentrations and transcription levels in SAP pigs.
Objective To observe the effects of immunologic cytokines or anti-angiogenesis gene transfer mediated by electroporation for choroidal melanoma cells.Methods The human embryo kidney cells and malignant choroidal melanoma cells were transfected with plasmids pNGVL-mIL2, pNGVL-mIL12, pCI-sFLK-1, pCR3.1-antiVEGF121,pCI-ExTek. Then the expression of mIL2, mIL12, sFLK-1, VEGF and ExTek were detected by enzymelinked immunosorbentassay (ELISA) and Western blot. Nude mice models of malignant choroidal melanoma were established and they were divided into four groups randomly. Each group was treated with 30 mu;l of 0.9% NaCl, 30 mu;g pNGVL, 30 mu;g antiVEGF121+sFLK-1+ExTek and 30 mu;g mIL2+mIL12 respectively by electroporation. Seven，14, 21, 28, 35 and 42 days after treatment, the tumor volumes were measured to calculate the tumor inhibition rate. Results ELISA and Western blot showed that mIL2，mIL12，sFLK-1 and ExTek were expressed after electroporation，VEGF expression was decreased remarkably. After treatment，the tumors of mIL2+mIL12 group were greatly inhibited with a tumor inhibition rate of 97.33%，while the tumors of antiVEGF121+sFLK-1+ExTek and pNGVL group were partially inhibited with tumor inhibition rates of 53.33% and 36.33% respectively.Conclusions Immunologic cytokines transfer mediated by electroporation can inhibit the growth of melanoma，but anti-angiogenesis only have a mild effects.
Objective To observe the levels of vascular endothelial growth factor (VEGF), interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1) in aqueous humor of patients with macular edema secondary to central retinal vein occlusion (CRVO). Methods Forty eyes of 40 consecutive patients with macular edema secondary to CRVO (CRVO group) were enrolled in this study. The patients included 25 males and 15 females. The patient age ranged from 38 to 76 years. The control group was 20 patients with senile cataract who underwent phacoemulsification, including 10 males and 10 females. The levels of VEGF165, VEGF165b, IL-6 and MCP-1 in aqueous humor were determined by enzymelinked immunosorbent assay. The correlation of VEGF, and IL-6, and MCP-1 were analyzed. Results The median aqueous level of VEGF165, IL-6 and MCP-1 were 1089.0, 165.6, 1253.0 pg/ml respectively in CRVO group, which were higher than the control group's results (168.2, 4.7, 216.4 pg/ml respectively), the differences were statistically significant (Z=-4.549, -6.008, -5.343;P＜0.001). The VEGF165b in CRVO group and control group were 834.0, 915.9 pg/ml respectively, the difference was not statistically significant (Z=-0.207，P＞0.05). The ratio of VEGF165b to VEGF165 in CRVO group and control group were 2.71, 7.28 respectively, the difference was statistically significant (t=-3.007，P＜0.05). There was a highly positive correlation between IL-6 and VEGF in CRVO group (r=0.526,P=0.001) and also mild positive correlation in control group (r=0.425，P=0.070). No correlation between MCP-1 and VEGF was observed in both groups (CRVO group: r=0.211,P＞0.05. Control group: r=-0.019,P＞0.05). Conclusions VEGF165, IL-6 and MCP-1 levels were increased in CRVO patients while the VEGF165b was normal. The ratio between VEGF165b and VEGF165 in aqueous humor of patients with macular edema secondary to CRVO was decreased.
Interleukin-1 (IL-1), interleukin-2(IL-2) and interleukin-6(IL-6) activities and tumor necrosis factor (TNF) contents in plasma from patients with different sites of cancers as well as controls using bioassay technique were studied. The results showed that the levels of IL-1,IL-2,IL-6 s from patients with different sites of cancer were decreased remarkably in comparision with controls and the contents of TNF from patients with different sites of cancers increased significantly. But the difference between different sites of cancer was not statistically significant. The data suggest that the variations in the contents of TNF and the levels of interleukins may be related to the development of these caner patients.
Objective To investigate the effects of cimetidine on the red cell immune function and interleukin-2(IL-2) in rats with obstructive jaundice. Methods Sixty SD rats were divided into bile duct ligation(BDL) group, cimetidine therapy (BDLC) group and sham operation(SO) group respectively. The red cell immue function and serum IL-2 level were determined with the red cell yeast-rosttes test and radioimmunoassay respectively. Results The red blood cell C3b receptor rosette rate(RBC-C3bRR), the red blood cell immune complex rosette rate(RICR), the red blood cell C3b receptor rosette-forming excited rate(RFER) and serum IL-2 level were significantly lower in BDL group as compared with SO group, the red blood cell C3b receptor rosette-forming inhibitory rate(RFIR) in BDL group was higher than that of SO group. After 7 days’ cimetidine therapy RBCC3bRR, RICR, RFER and IL-2 became higher than those of BDL group, but RFIR was lower than that of BDL group. Conclusion Supplemental cimetidine can significantly enhance the impaired red cell immune function and IL-2 production in rats with obstructive jaundice.
Objective To compare the surgical trauma between videoassisted thoracoscopic surgery(VATS) and conventional thoracotomy, and to investigate the possible minimally invasive mechanism. Methods Seventyseven patients who had undergone consecutive operations from April 2005 to January 2006 were chosen from cardiothoracic surgery department of Fujian Provincial Hospital. Twentytwo cases had spontaneous pneumothorax diagnosed by chest X-ray examination, twentynine had patent ductus arteriosus diagnosed by color echocardiography, and twentysix had congenital atrial septal defect. According to lesions and operative methods, the patients were divided into two groups: conventional thoracotomy group(CTH group) and videoassisted thoracoscopic surgery group(VATS group). The concentrations of serum C-reactive protein（CRP），interleukin6 (IL-6)，interleukin-8（IL-8） and tumor necrosis factor-α(TNF-α) were selected as indexes to measure surgical trauma. ARRAY360 specific protein and pharmaceutical analysis system were used to determine CRP automaticly at the day before operation and on the 1st, 2nd and 3rd day after operation. Radioimmunoassay was used to measure the concentrations of IL-6，IL-8 and TNF-α. Clinical indexes such as operative time, cardiopulmonary bypass (CPB) time, intraoperative blood lost, postoperative analgesic time and hospitalization time were analyzed and compared. Results Under the condition that patients had the same diseases, there was no statistical significance in preoperative concentrations of serum CRP，IL-6，IL-8 and TNFα between VATS group and CTH group（P=0.067, 0.062, 0.053,0.064）. The concentrations of serum CRP(P=0.045,0.043,0.044), IL-6(P=0.042，0.032，0.039), IL-8(P=0.046，0.045，0.048) and TNF-α(P=0.041，0.043，0.043) on the 1st, 2nd and 3rd day after operation were significantly lower in VATS group than that in CTH group (Plt;0.05). Compared with CTH group, there were less blood lost（P=0.032）, shorter postoperative analgesic time and hospitalization time（P=0.041） in VATS group. There was no statistical significance in CPB time between two groups. However, hospitalization time varied with different diseases. Conclusion Compared with conventional thoracotomy，videoassisted thoracoscopic surgery has less surgical trauma, less intraoperative blood lost, shorter postoperative analgesic time, and can make patients recover rapidly. So it is worth spreading.
ObjectiveTo observe the effect of Crocin on structure and the expression of tumor necrosis factor-alpha; (TNF-alpha;) and interleukin-1beta; (IL-1beta;) in rat retina after injury by ischemia-reperfusion. Methods A total of 80 Sprague-Dawley male rats at the age of 8 -10 weeks were divided into control group, model group, low-dose Crocin group and high-dose Crocin group, with 20 rats in each group. The rats of control group were not treated. The rats in model, low-dose Crocin and high-dose Crocin group were induced with normal saline by anterior chamber perfusion creating a retinal ischemia-reperfusion (RIR) model. The rats of the low-dose Crocin and highdose Crocin group received intraperitoneal injection with different doses of Crocin solution (5 mg/kg, or 50 mg/kg) 30 minutes prior to ischemic injury and one time per day after successful RIR. Optical microscopy was used to observe the retinal structure. Enzymelinked immunosorbent assay (ELISA) was used to measure the expression of TNF-alpha; and IL-1beta; 6, 12, 24 and 48 hours after RIR. ResultsThe retinal structure of control group was normal. Pathological changes were found in the RIR model and low-dose Crocin group, such as retinal edema, disorganized structure and loosely packed cells. The degree of pathological changes in lowdose Crocin group was less than the RIR model group. The retinal structure of high-dose Crocin group was similar to the control group. The expression of TNF-alpha; was the highest at 24 hours after modeling, while the expression of IL-1beta; was the highest at 12 and 48 hours after RIR modeling. Six, 12, 24 and 48 hours after RIR modeling, compared with the control group, the TNF-alpha; expression of model (t=5.42, 7.94, 9.32, 9.18;P＜0.05 ), low-dose Crocin (t=3.94, 4.12, 4.98, 3.84;P＜0.05) and high-dose Crocin group (t=2.13, 2.34, 2.96, 2.78;P＞0.05) were increased. Compared with the RIR model group, the TNF-alpha; expression of low-dose Crocin (t=3.95, 4.56, 4.01, 5.12) and high-dose Crocin group (t=5.23, 7.65, 7.74, 7.63) was decreased. Compared with the control group, the IL-1beta; expression of model (t=7.23, 7.87, 7.15, 15.60), low-dose Crocin (t=5.65, 5.10, 5.54, 6.87;P＜0.05) and high-dose Crocin group (t=4.38, 5.21, 4.56, 4.75) was increased (P＜0.05). Compared with the model group, the IL-1beta; expression of low.dose Crocin group was decreased significantly 48 hours after RIR modeling (t=7.56,P＜0.05); but it decreased significantly at each time point in high-dose Crocin group (t=6.94, 5.36, 6.05, 10.50;P＜0.05). Conclusion Crocin can improve the retinal pathologic changes, while down-regulating TNF-alpha; and IL-1beta; expression in RIR rats.
Objective To observe the serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), platelet 5-HT and blood platelet count, emotion and burn injury healing of patients with moderate and severe burn injury and anxiety-depression symptoms. Methods In-patients with moderate and severe burn injury were selected from 2003.4 to 2005.2 and then divided into anxiety-depression group and control group according to their anxiety-depression scores by Hamilton Rating Scale for Depression (HAMD ) and Hamilton Rating Scale for Anxiety (HAMA) 3 days after being burnt. Routine therapy was given to two groups, which lasted 1 month. Their scores of anxiety and depression and the degree of injury healing were observed, and the serum levels of TNF-α and IL-6, platelet 5-HT and blood platelet count were measured in the two groups. Results Fifty-one in-patients with moderate and severe burn injury were divided into the anxiety-depression group (24 cases) and the control group (27 cases). After 30-day treatment, the depression scores did not decrease in the anxiety-depression group (P=0.12), but the anxiety scores decreased (P=0.00). In the anxiety-depression group, the burn injury healing time was postponed (P=0.00), the serum levels of TNF-α increased (P=0.00), and the platelet 5-HT levels decreased (P=0.04) before and after treatment. Conclusion Depressive reaction occurs in patients with moderate and severe burn injury, which is a continuously negative emotion. It can lead to high levels of serum TNF-α, reduction in platelet 5-HT, and delayed burn injury healing time. Due to the limited sample size and different location of patients, there may be some bias in this conclusion. We are prepared to increase the sample size and select patients in the same region in further relevant studies.
ObjectiveTo investigate the inhibitory effects of L arginine (L arg) on systemic inflammatory response after cardiopulmonary bypass(CPB).MethodsFifty one patients with rheumatic heart disease were randomly divided into two groups: L arg group ( n =25) and control group ( n =26). For L arg group, L arg at 300mg/kg was given during operation. Plasma levels of tumor necrosis factor α(TNF α),interleukin 1β(IL 1β)and interleukin 10(IL 10) were measured by enzyme linked immunosorbent assay technique at baseline(before operation) and at 2,4,8,24 and 48 h after CPB termination.ResultsTNF α,IL 1β and IL 10 levels were increased in both groups after CPB ( P lt;0.05); levels of TNF α, IL 1β returned to normal at 48 h after CPB; In L arg group, TNF α and IL 1β levels were significantly lower than those in control group at 4,8 and 24 h after CPB ( P lt; 0 05). No significant difference were detected in IL 10 between groups( P gt;0.05).ConclusionL arg may decrease plasma levels of TNF α and IL 1β after CPB, it implies L arg may inhibit inflammation induced by CPB.
Objective To investigate the expressions of ubiquitin-proteasome markers，including E2-14K，MAFbx，MuRF-1，and nuclear factor-κB（NF- κB） p50，in diaphragm of COPD rats，and their relationship with IL-17 level in diaphragm and serum in order to elucidate the potential mechanism of diaphragm atrophy. Methods Thirty healthy adult male SD rats were randomly divided into a model group （n=18） and a normal control group （n=12）. The COPD rat model was established by instillation of lipopolysaccharide （LPS） and exposure to cigarette smoke for 28 days. The protein levels of E2-14K，MAFbx，MuRF-1，and NF-κB p50 in diaphragm were measured by Western blot. The concentration of IL-17 in serum and diaphragm was measured by ELISA. Results Western blot showed that the protein expressions of E2-14K，MAFbx，MuRF-1，and NF-κB p50 in diaphragm increased significantly in the COPD model group compared with the normal control group （0.96±0.12 vs. 0.53±0.09，0.99±0.10 vs. 0.53±0.08，0.95±0.08 vs. 0.51±0.16，1.11±0.10 vs. 0.64±0.50，respectively，Plt;0.01）. The IL-17 level in serum and diaphragm was significantly higher in the COPD group than the control group. The expression of NF-κB p50 was positively correlated with E2-14K，MAFbx，and MuRF-1 expressions （r=0.82，0.92，0.86，respectively，Plt;0.01）. Both in serum and diaphragm，IL-17 level was positively correlated with the percentage of neutrophils，levels of NF-κB p50，E2-14K，MAFbx，and MuRF-1 expressions（all Plt;0.01）. The IL-17 levels in serum and diaphragm were also positively correlated each other （r=0.84，Plt;0.01）. Conclusions The results show that the ubiquitin-proteasom pathway，the NF-κB pathway and IL-17 are up-regulated in diaphragm of COPD rats .These alterations may contribute to diaphragm atrophy in COPD.