Effect of Wumeiwan on Cytokines TNF-α, IL-6, IL-8, IL-10 and Expression of NF-κBp65 in Rats with Ulcerative Colitis

The effects of Wumeiwan （WMW） on TNF-α, IL-6, IL-8, IL-10 and NF-κBp65 in rats with ulcerative colitis （UC） were investigated, the curative effectiveness of WMW vs salicylazosulfapyridine （SASP） was compared, and the action mechanism was analyzed. Fifty-Six Sprague-Dawley （SD） rats were randomly divided into four groups （n=14 in each group, with equal ratio of male and female）： normal control group, model group, SASP group, and WMW group. Except normal control group, the rat UC models in the remaining three groups were established using the method of 2.4-dinitrochlorobenzene （DNCB） immunization and acetic acid local enema. The rats in model group, SASP group, and WMW group were treated with distilled water, SASP, and WMW respectively. The changes in the symptoms and signs were observed, and levels of IL-6, IL-8, TNF-α, IL-10 and the expression of NF-κBp65 in the colonic tissues were statistically analyzed. The results showed that the levels of IL-6, IL-8, and TNF-α were significantly increased （P〈0.01）, while those of IL-10 significantly reduced （P〈0.01） after establishment of rat UC models as compared with normal control group. The levels of IL-6, IL-8, and TNF-α were obviously lower, but the level of IL-10 was obviously higher in WMW and SASP groups than those in model group （P〈0.05）. The levels of IL-6, IL-8, and TNF-α were lower, while the level oflL-10 was higher in WMW group than in SASP group. NF-κBp65 was expressed negatively or weakly in normal colonic tissues. The positive expression rate of NF-κBp65 in WMW group and SASP group was obviously lower than in model group （P〈0.01）, and there was significant difference between WMW group and SASP group （P〈0.05）. It was concluded that rat UC model was established successfully. WMW could up-regulate the expression of IL-10, down-regulate the expression of TNF-α, IL-6, IL-8, and inhibit the NF-κBp65 activity to adjust immune function, indicating WMW had better curative effects on UC in rats.     

Expression of IL-βα and TNF-α in MCMV Myocarditis and Its Role

In order to study the expression of IL-β and TNF-α in the myocardium of MCMV myocarditis and their role in the myocardial damages, 60 BALB/C mice of 4 weeks were randomly divided into two groups: 36 were injected intraperitoneally with MCMV and 24 served as control group. Immunohistochemistry was used to detect IL-β and TNF-α expression in the myocardium, and myocardial lesions were observed histopathologically. Histopathological study on the myocardium from infected mice revealed focal or diffuse lesions characterized by inflammatory cells and degeneration or necrosis of myocytes. The myocardial lesion score showed the degree of inflammatory cell infiltration was slight in MCMV myocarditis. The positive staining signals for IL-β and TNF-α proteins which mainly located in the infiltrating inflammatory cells and degenerative or necrotic myocytes were markedly detectable whereas there were no positive findings in the myocardium of control mice. IL-β and TNF-α was expressed in the myocardium of viral myocarditis murine model induced by MCMV. IL-β and TNF-α may play an important role in the pathogenesis of viral myocarditis.     

Expression of IL-1β and TNF-α in MCMV Myocarditis and Its Role

Autoi mmune myocarditis in Coxsackievirus B3(CB3)-infected mice is associated with infiltrationof the heart by inflammatory cells that secreteTNF-αand IL-1[1].In this study we examine theexpression of cytokines , TNF-αand IL-1βpro-duced locallyinthe heart inresponse to MCMVin-fectionin order to determine their potential role inthe development of acute myocarditis .1MATERIALS AND METHODS1 .1Reagents and AntibodiesMonoclonal goat anti-mouse IL-1β, TNF-αanti-body and SABC …     

Responses of CRP, TNF-α and IL-6 in minimally invasive esophagectomy and open esophageal resection

目的 通过对比研究了解微创食管癌手术与传统开放术式术后C反应蛋白( C-reactive protein,CRP)及细胞因子水平变化.方法 选择2009年1月至2011年2月在重庆医科大学第一附属医院胸外科接受微创食管癌切除术(minimally invasive esophagectomy,MIE)与开胸食管癌切除术(open esophageal resection,OPEN)的患者各10例,分别于术前,术后4、24、48、72、120、168 h抽取静脉血标本,用双抗体夹心ABC-ELISA法测定TNF-α、IL-6,用超敏CRP分析测定CRP值.结果 2组人群术前、术后血浆TNF-α水平差异没有统计学意义,2组术后4、24、120 h血清CPR水平差异有统计学意义(P＜0.05),2组术后各时间点血清IL-6水平差异均有统计学意义(P＜0.05);两因素重复测量资料的方差分析提示MIE组和OPEN组术后不同时间点血清IL-6细胞因子浓度差异有统计学意义(P＜0.01),且手术方式与时间因素之间存在交互作用.结论 微创食管癌手术与传统食管癌手术相比,可能会减少术后机体炎症因子释放,减少术后炎症反应.     

Study on Cytokines IL-2,IL-6,IL-10 in Patients of Chronic Allergic Rhinitis Treated with Acupuncture

To observe the plasmatic concentration of IL-6,IL-10 and IL-2 in the patient of chronicallergic rhinitis before and after acupuncture therapy.Methods:Cytokine levels were determinedbefore and after treatment in 30 healthy volunteers (Group A) and 90 patients of chronic allergicrhinitis (Group B) with an increased plasma IL-10 level.Group B was then divided into 3 subgroups:30 patients treated with real acupuncture (Group B1);30 patients treated with sham acupuncture(Group B2);30 non-treated patients (Group B3).Results:The allergic subjects of group B1,compared with controls,showed a significant reduction of IL-10 after a specific treatment withacupuncture (P<0.05).On the other hand,in those patients treated with sham acupuncture (B2) aswell as in non-treated patients (B3),the IL-10 values remained high and unchanged.There was astatistically significant change in IL-2 values at 24 hours (P<0.05) after real acupuncture (Groups A,B1),however the values remained within normal ranges.The IL-6 do not change after therapy.Conclusion:The acupuncture treatment can reduce plasmatic level of IL-10 in chronic allergicrhinitis.     

Impairment of Myocardial and Skeletal Mitochondria in Mice with Viral Myocarditis and Their Correlation

In order to investigate the impairment of mitochondrial membrane phospholipid local- ization and DNA3867 (mtDNA3867) deletion and the correlation between cardiac and skeletal muscle cells in mice with viral myocarditis, 50 BALB/c mice were divided into two groups randomly. In ex- perimental group (n=40), the mice were intraperitoneally injected with 0.1 mL Eagle liquid with CVB3 (TCID50=108), while in the control group (n=10), the mice were subjected to equal volume of Eagle liquid. The impairment of mitochondrial membrane phospholipid localization and mtDNA3867 deletion rate of cardiac and skeletal muscle were detected separately at day 3, 11 and 24 after injec- tion. The correlation of mitochondrial membrane phospholipid localization and mtDNA3867 deletion rate between cardiac and skeletal muscle cells cells was analyzed using Spearman method. At the day 3 after injection, in both cardiac and skeletal muscle cells, mtDNA3867 deletion rate was significantly higher in experimental group than in control group (P<0.05), but the localization of mitochondrial membrane phospholipid showed no difference between two groups (P>0.05). At day 11 after injec- tion, the mtDNA3867 deletion rate of both cells in experimental group was increased to the peak level (P<0.05), and the impairment of mitochondrial membrane phospholipid localization of both cells also increased markedly in experimental group as compared with control group (P>0.05). At the day 24 after injection, the impairment of mitochondrial membrane phospholipid localization and mtDNA3867 deletion of both cells showed a recovery tendency, but still severer than those at the day 3 after injec- tion (P<0.05). The impairment of mitochondrial membrane phospholipid localization and mtDNA3867 deletion were consistent and synchronistic between cardiac and skeletal muscle cells, and showed good correlations (P<0.05). The impairment of mitochondria plays an important role in the patho- genesis of viral myocarditis, and the skeletal muscle cells might act as a peripheral "window" to re- flect the mitochondrial damage of cardiac myocytes.     

TNF-α and IL-8 of the Patients with Allergic Asthma

The levels of serum TNF-α and IL-8 in the patients with allergic asthma during acute attack period and remission period, and the effects of glucocorticoid (GC) on them were investigated. By using ELISA, the levels of TNF-α and IL-8 were detected in the healthy volunteers (group C, n=40), the patients with allergic asthma (n=40) during acute attack period (group A) and remission period (group B) and those taking GC for a week (n=28). The results were compared among them. It was found that the levels of TNF-α and IL-8 in group A were higher than in group B and group C. In the patients subject to GC therapy, the levels of TNF-α and IL-8 were decreased as compared with those in group A. In group B, the level of TNF-α was higher than in group C, but there was no significant difference in the level of IL-8 between group B and group C. It was concluded that the inflammatory cytokines, TNF-α and 11.-8, played important roles in the bronchus allergic inflammation. GC could reduce the levels of serum TNF-α and IL-8 to exert the anti-inflammatory effects.     

Expression of urotensin Ⅱ and its relationship with pro-inflammatory cytokines of TNF-α and IL-1β in early stage of acute liver failure

目的 探讨小鼠急性肝衰竭(ALF)早期尾加压素Ⅱ(UⅡ)表达和分泌的动态变化及其对前炎症细胞因子的影响.方法 选择6周龄雄性BALB/c小鼠60只,随机分为模型组和预处理组,每组30只.两组均建立ALF动物模型,即采用脂多糖(LPS)50 μg/kg联合右旋半乳糖胺(D-GalN )800 mg/kg(LPS/D-GalN)腹腔注射;预处理组在LPS/D-GalN注射前0.5h,以UⅡ受体拮抗剂urantide 0.6 mg/kg尾静脉注射.两组均在LPS/D-GalN注射后0、0.5、1、2和6h处死动物(每一时间点各6只小鼠),其中0h作为对照(仅腹腔内注射0.2 mL无菌生理盐水);采集动物血清和肝组织标本.采用RT-PCR及ELISA方法分别检测U Ⅱ、肿瘤坏死因子α(TNF-α)及白介素1β(IL-1β)mRNA和蛋白质的表达.结果 LPS/D-GalN注射后0.5h,肝组织和血清中UⅡ的表达和分泌即快速上升并达峰值,且峰值水平持续至LPS/D-GalN攻击2h后,至6h时UⅡ的表达与分泌虽有所降低,但仍显著高于正常水平(p＜0.05);面TNF-α水平在LPS/D-GalN攻击后0.5h内无明显升高(P＞0.05),直到1h后才快速上升达峰值(P＜0.05),至6h时开始下降(P＜0.05);IL-1β的表达与分泌则直到6h后才有明显上升(P＜0.05).Urantide的应用不仅显著抑制了LPS/D-GalN攻击诱导的UⅡ高表达,也使上调的TNF-α和IL-1β表达和分泌明显受抑(P＜0.05).结论 UⅡ可刺激TNF-α的表达,有可能作为炎症级联效应的触发者在ALF的发生或启动中发挥关键性影响.     

Clinical significance of detecting serum levels of TNF-α, IL-10, IL-8 and IL-6 in patients with Breast cancer

目的 探讨乳腺癌患者血清中IL--6、IL-8、IL-10和TNF-α水平与乳腺癌发病的关系.方法 应用放射免疫分析方法检测50例乳腺癌患者不同时期血清IL-6、IL-8、IL-10和TNF-α水平,并与30例正常者对照比较.结果 乳腺癌患者活动期血清中IL-6、IL-8和TNF-α的含量显著高于缓解期和对照组,IL-10含量显著低于缓解期和对照组.缓解期IL-6、IL-8、IL-10和TNF-α含量与对照组比较差异无统计学意义(P＞0.05).活动期IL-6、IL-8表达水平与TNF-α呈正相关,与IL-10呈负相关.结论 乳腺癌患者活动期IL-10表达水平降低,IL-6、IL-8和TNF-α 表达水平升高,使体内免疫调节失衡,从而成为乳腺癌患者病因之一.乳腺癌患者血清中IL-6、IL-8、IL-10和TNF-α水平与疾病严重程度显著相关.     

Correlation between the Expression of Fas, Bcl-2 in Peripheral Blood Lymphocytes and the LeveL of IFN-γ,IL-4 in Setum of Patients with Condyloma Acuminata

Summary In order to investigate the correlation between the expression of the apoptotic regulatory proteins (Fas, Blc-2) in peripheral blood lymphocytes (PBLC) and the level of IFN-γ and IL-4 in serum of the patients with condyloma acuminata (CA) in the immune pathogenesis of CA, indirect immunofluorescence labeling method of flow cytometer and solid sandwich ELISA method were performed for detecting the expression of Fas, Bcl-2 in PBLC and the level of IFN-γ and IL-4 in serum of 60 cases of CA. The results showed the expression level of Fas in PBLC of CA was significantly higher than in the normal control group, but the expression level of Bcl-2 was significantly lower (bothP<0.01). The level of IFN-γ in serum of CA was significantly lower than in the normal control group (P<0.01), but IL-4 was significantly lower (bothP<0.01). The expression of Fas in PBLC had a negative correlation with the level of IFN-γ in serum of patients with CA, but had a positive correlation with the level of IL-4: The expression of Bcl-2 had a positive correlation with the level of IFN-γ, but had a negative correlation with the level of IL-4. All the correlation coefficients had significant differerce byt test (P<0.01). It was suggested abnormal apoptosis in PBLC, the suppressed secretion of the TH1-associated cytokines (eg: IFN-γ) and the increased secretion of the TH2-associated cytokines (eg: IL-4) existed in the patients with CA and might play an important role in the immune pathogenesis of CA. LIU Houjun, male, born in 1963, Associate Professor     

Significance and Expression of Serum and Glucocorticoid-inducible Kinase in Kidney of Mice with Diabetic Nephropathy

To investigate the expression and the role of three isoforms of Serum and Glucocorticoidinducible Kinase (SGK) in experimental diabetic nephropathy (DN), 12 male C57BL/6 mice of 8-weeks-old were divided into two groups. Streptozotocin (STZ)-induced diabetic nephropathy and normal controls were analyzed at the end of the 4th week after the induction of diabetes. Renal hemodynamics and histological studies were performed. The expression of SGK1 mRNA, SGK2 mRNA and SGK3 mRNA of kidney cortex were measured by RT-PCR, and the cortical SGK1 protein was detected with Western blotting. Our results showed that the blood glucose, blood HbA1c, 24h urinary protein, creatinine clearance and the renal index were all increased in DN group. More extracellular matrix (ECM) accumulation was observed. The level of cortical SGK1 mRNA and protein were up-regulated in DN group in comparison with control group. SGK2 and SGK3 mRNA were elevated in DN mice. In DN, mRNA level of three SGK isoforms and SGK1 protein were increased significantly. It is concluded that SGKs may contribute to the early renal injury of DN.     

Expression of cyclooxyenase-2 protain and its relationship with HIF-1α in HCC

Objective： To investigate the clinical significance of COX-2 （Cyclooxygenase-2） expression in HCC （Primary hepatocellular carcinoma） and clarify whether COX-2 is correlated with hypoxia-inducible factor-1α （HIF-1α） in the development of HCC. Methods： Tumor tissues were obtained from 53 patients with HCC. COX-2 and HIF-1α were determined by immunohistochemistry. All 53 patients were regularly followed up and the data were collected prospectively. Results： Immunostaining showed the expression of COX-2 （ n = 33, 62.3 % ） and HIF-1α （ n = 36, 67.9% ） in most tumor cells. The level of COX-2 was correlated with HIF-1α levels（ r = 0.4413, P 〈0.01 ）. There were significant correlation between clinicopathological features and higher tumor cytosolic COX-2 level was in the presence of multiple tumors （ P = 0.01）, venous invasion （ P = 0.03）, advanced tumor stage （ P = 0.01）, and well-different tumor grade （P =0.03）. High-tumor cytosolic COX-2 level was correlated with patient＇s worse prognosis （P = 0.0085）. Conclusion： Elevated tumor COX-2 level is correlated with elevated HIF-1α levels and invasiveness in HCC, suggesting COX-2 plays an important role in the progression of HCC, and may be an important therapeutic target in HCC.     

Plasma Levels of the Anti-inflammatory Cytokine IL-10 and Inflammatory Cytokine IL-6 in Patients with Unstable Angina

The plasma levels of inflammatory cytokine interleukin-6 （IL-6） and anti-inflammatory cytokine interleukin-10 （IL-10） in the patients with unstable angina or stable angina were determined and compared. In 30 patients with unstable angina and 22 patients with stable angina, plasma levels of IL-10 and IL-6 were detected by ELISA and plasma lipid parameters by lipid research clinical methods respectively. The results showed plasma levels of IL-10 were significantly lower in unstable angina group than in stable angina group （P=0. 005）, while those of IL-6 were significantly increased in unstable angina group as compared with those in stable angina group （P= 0. 039）. There was a significantly negative correlation between IL-10 and IL-6 in patients with unstable angina （r=-0.41, P=0. 003）. In the unstable angina group, IL-6 levels were obviously positively correlated with TC （r=0. 314, P=0. 023）, but not with TG and HDL. There were no significant correlations between IL-10 and plasma lipid parameters. It was suggested that the decreased IL-10 and increased IL-6 might be associated with the atheromatous plaque stability and progression of coronary heart diseases. IL-10 may play an important role in preventing coronary vascular lesions.     

Tc1/Tc2 Imbalance in the Peripheral Blood of Patients with Recurrent Genital Herpes

In order to investigate the IFN-γ and IL-4 expression of CD8^＋T lymphoeytes in the peripheral blood from patients with recurrent genital herpes （RGH） at different clinical periods and their relationship with the pathogenesis of RGH, flow cytometry was used to detect the intracellular cytokines （IFN-γ and IL-4） of CD8^＋ T lymphocytes in the peripheral blood of 30 patients with RGH at acute period, 20 patients with RGH at recovery period and 15 healthy volunteers. The results showed that RGH patients at acute period had a lower percentage of Tcl subsets in peripheral blood than that of healthy controls （P〈0. 001）, especially a remarkable decreased percentage of Tc1 subsets （P〈0. 001） among those RGH patients with recurrent number more than 3 in the recent half a year. Tc1/Tc2 ratio in the RGH patients at acute period was significantly decreased as conapared with normal control group （P〈0.05）. The recurrent number of acute patients in the recent half a year was significantly correlated with the percentage of Tc1 subsets and the ratio of Tc1/Tc2 （P〈 0.05）. A decreased percentage of Tc1 subsets was found among the RGH patients with recurrent number more than 3 in the recent half a year at recovery period in comparison with healthy volunteers （P〈0.05）, and it was significantly correlated with the recurrent number in the recent half a year （P〈0.05）. It is concluded that there are Tc1/Tc2 imbalance and a low level of Tc1 subsets in RGH patients who are relapsing repeatedly in the near period. The low level of Tc1 subsets may be an important factor for the recurrence of RGH and the reactivation of latent herpesvirus infection.     

The Expression of TNF-α and ICAM-1 in Lesions of Lichen Planus and Its Implication

In order to investigate the role of TNF-α and ICAM-1 in the pathogenesis of lichen planus, immunohistochemistry was used to detect the expression of TNF-α and ICAM-1 in skin le- sions of the patients with lichen planus and skin tissues of normal subjects. The results showed that positive rates of TNF-α and ICAM-1 expressions in lichen planus were significantly higher than those in normal skins (both P<0.05). Meanwhile, there was a obvious correlation between the in- crease of TNF-α and that of ICAM-1 in lichen planus. The expression of TNF-α and ICAM-1 might play an important role in the development of lichen planus.