Relationship between circulating serum soluble interleukin-6 receptor and the angiogenic cytokines basic fibroblast growth factor and vascular endothelial growth factor in multiple myeloma

Angiogenesis plays an important role in multiple myeloma (MM) progression. Various mitogens such as vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (FGF-2) have been implicated in the angiogenic process of various malignancies. Interleukin-6 (IL-6) is a growth factor of myeloma cells and its signaling is mediated via a cell surface receptor complex (IL-6r). IL-6 and tumor necrosis factor-alpha (TNF-alpha) are involved in the secretion of VEGF by IL-6r expressing myeloma cells. In this study, serum FGF-2, VEGF, IL-6r, and TNF-alpha were measured in 46 untreated MM patients and were studied in relation to disease stage (by Salmon-Durie criteria) and severity [assessed by serum beta(2)-microglobulin (beta(2)M), C-reactive protein (CRP), alpha(1)-antitrypsin (alpha(1)AT), and lactic dehydrogenase (LDH) levels]. The results showed that FGF-2, VEGF, IL-6r, and TNF-alpha were significantly elevated in MM patients in comparison to controls ( p<0.008) and were significantly higher in stage III disease in comparison to stages I and II ( p<0.03). The mean concentrations of IL-6r were 877+/-374, 1220+/-308, 1431+/-878, and 453+/-180 pg/ml for stages I, II, and III and controls, respectively. Levels of beta(2)M, alpha(1)AT, CRP, and LDH were all significantly higher in MM patients than controls and increased with advancing stage of disease. There were positive correlations of both VEGF and FGF-2 with IL-6r, TNF-alpha, beta(2)M, alpha(1)AT, CRP, and LDH. We conclude that IL-6r and TNF-alpha increase in parallel to VEGF and FGF-2 with increasing stage of MM disease. These molecules correlate with biochemical markers of disease activity and may play a role in the progression of multiple myeloma.     

胰腺癌组织中p53、血管内皮生长因子的表达与血管生成的关系及其临床意义

目的 探讨胰腺癌组织中p53、血管内皮生长因子(VEGF)和血管生成的关系。方法 用免疫组织化学方法检测48例胰腺癌组织及癌旁组织、6例正常胰腺组织中p53、VEGF表达和微血管密度(MVD)。结果 胰腺癌组织中VEGF、p53的阳性表达率分别为54.17％和50％,显著高于癌旁组织及正常组织的表达率(P<0.01),胰腺癌组织中MVD显著高于癌旁组织及正常组织。VEGF表达与肿瘤大小和分期有关(P=0.038,P=0.045),VEGF表达与MVD有相关性(r=0.294 P=0.043)。p53与淋巴结转移及预后相关(P<0.05)而与VEGF、MVD之间无关。MVD与胰腺癌临床病理特征无关,MVD与生存期存在负相关(r=0.371 P=0.011)。多元回归分析显示p53、VEGF和MVD都不是影响胰腺癌预后的独立因素。结论 p53基因突变为胰腺癌分子事件的晚期事件,可作为评价胰腺癌预后的一项指标,抗血管生成可能有利于胰腺癌的治疗。     

IgG antibodies to type II collagen reflect inflammatory activity in patients with rheumatoid arthritis

OBJECTIVE: To determine the clinical significance of IgG antibodies to type II collagen (CII) and to define any correlation of antibodies to CII with the inflammatory response in patients with rheumatoid arthritis (RA). METHODS: IgG antibodies to native human type II collagen (IgG anti-CII) were measured in sera and synovial fluid (SF) from patients with RA, patients with osteoarthritis (OA), and healthy controls by an improved ELISA. Demographic, clinical, and laboratory data including tumor necrosis factor-alpha (TNF-alpha) and interleukin 6 (IL-6) levels were also obtained at the time of sampling in patients with RA. RESULTS: The median level and positivity for circulating IgG anti-CII were higher in patients with RA (n = 297) than patients with OA (n = 34) and healthy controls (n = 50) (p < 0.001). The titers of IgG anti-CII in SF were also higher in RA (n = 45) than in OA (n = 16) (p < 0.001). In paired samples, the levels of IgG anti-CII were significantly higher in SF compared to the sera in patients with RA (n = 45) (p < 0.001), but levels were not different in patients with OA (n = 16). Circulating IgG anti-CII converted from positive to negative in 13 patients (10.7%) and from negative to positive in 18 patients (14.8%) among 122 patients with RA in whom IgG anti-CII were monitored sequentially at a mean interval of 12.2 months. IgG anti-CII positive patients (n = 98) had shorter disease duration (p = 0.04) and less frequent deformity (p = 0.013), and higher median erythrocyte sedimentation rate (ESR) (p = 0.001) and C-reactive protein (CRP) (p < 0.001) than IgG anti-CII negative patients (n = 120). The levels of IgG anti-CII correlated with CRP (r = 0.270) and ESR (r = 0.253). CRP decreased significantly in patients (n = 13) who converted from IgG anti-CII positive to negative (p = 0.013). IgG anti-CII positive patients (n = 40) had higher levels of TNF-alpha and IL-6 than negative patients (n = 40) (p < 0.001). Levels of IgG anti-CII correlated well with TNF-alpha (r = 0.617) and IL-6 (r = 0.347). CONCLUSION: Increased IgG anti-CII in sera and SF in RA correlated directly with acute phase reactants and the proinflammatory cytokines TNF-alpha and IL-6. Our data suggest that IgG anti-CII could reflect inflammatory activity with a potential to destroy cartilage in the early stages of RA.     

Relationship between cytokines and tumour markers in patients with chronic heart failure

AIMS: Serum levels of some cytokines and tumour markers are elevated in patients with chronic heart failure (HF). We aimed to investigate the relationship between circulating levels of cytokines and tumour markers in patients with HF. METHODS: We included 35 HF patients and 33 normal controls. HF patients were divided into two groups: mild HF (NYHA class I/II) (n=10) and severe HF (NYHA class III/IV) (n=25). Serum cytokine levels (TNF-alpha, IL-1 beta, IL-6, and IL-10) were measured by ELISA and tumour markers (CA 125, CA 19-9, CA 15-3, CEA and AFP) by chemiluminescent enzyme immunoassay. RESULTS: Serum levels of TNF-alpha, IL-6, and IL-10 as cytokines, and CA 125 and CA 19-9 as tumour markers were significantly higher in HF patients than in normal controls (p<0.0001 for all). Serum levels of TNF-alpha, IL-6 and IL-10 and CA 125 in the severe HF patients were significantly higher than in the mild HF patients (p<0.001 for all). Correlation analysis showed that CA 125 was positively related to TNF-alpha (r=0.624, p<0.001), IL-6 (r=0.671, p<0.001), and IL-10 (r=0.545, p<0.001) in HF. CONCLUSION: These findings show that CA 125 is markedly elevated in patients with HF, and correlates with serum TNF-alpha, IL-6 and IL-10 levels. Therefore, we speculate that among the tumour markers studied, only CA 125 is closely related to the cytokine system.     

The clinical and biological effects of thalidomide in patients with myelodysplastic syndromes

Thirty patients with myelodysplastic syndromes (MDS) were treated with thalidomide at 100 mg/d p.o., increased as tolerated to 400 mg/d for 12 weeks. Levels of apoptosis, macrophage number, microvessel density (MVD), tumour necrosis factor alpha (TNF-alpha), transforming growth factor beta (TGF-beta), interleukin 6 (IL-6), vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were determined in the serum, bone marrow (BM) plasma and BM biopsies before and after therapy. Pretherapy biological characteristics of MDS patients were compared with similar studies performed in 11 normal volunteers. Ten patients demonstrated haematological improvement in the erythroid series, six becoming transfusion independent. Responders had a higher pretherapy platelet count (P < 0.048) and lower BM blasts (P < 0.013). Median time to response was 10 weeks, and four remain in remission beyond a year. Pretherapy MDS BMs showed higher MVD (P < 0.001) and TGF-beta (P < 0.03) and higher serum TNF-alpha (P < 0.008) compared with normal control subjects. After therapy, only BM TGF-beta decreased significantly (P < 0.002). Pretherapy haemoglobin was directly related to serum VEGF (P < 0.001) in responders and inversely related in non-responders (P < 0.05), suggesting the possibility that angiogenesis may be a primary pathology in the former and a consequence of anaemia-induced hypoxia in the latter. We conclude that thalidomide has important clinical and biological effects in at least a subset of MDS patients, but the precise mechanism of its action remains unknown and requires further study including a larger number of patients.     

Proinflammatory cytokines, transforming growth factor-beta1, and fibrinolytic enzymes in loculated and free-flowing pleural exudates

STUDY OBJECTIVES: To measure tumor necrosis factor (TNF) alpha, interleukin (IL) 1beta, and transforming growth factor (TGF) beta1 in loculated and free-flowing pleural effusions caused by malignancy, tuberculosis (TB), and pneumonia and their relationship with plasminogen activator inhibitor-type 1 (PAI-1) and tissue-type plasminogen activator (tPA) and to compare the differences between loculated and free-flowing effusions. DESIGN: A prospective study. PATIENTS AND METHODS: The effusion levels of TNF-alpha, IL-1beta, TGF-beta1, PAI-1, and tPA were measured in 29 patients with malignant effusions, 19 patients with TB, and 30 patients with parapneumonic effusions. Pleural effusions were divided into loculated and free-flowing groups by imaging studies. A group of 42 patients with loculated effusions was subdivided into primary and secondary loculation groups by chest ultrasonography. RESULTS: The median levels of TNF-alpha (87.0 pg/mL), IL-1beta (13.8 pg/mL), TGF-beta1 (10,952.9 pg/mL), PAI-1 (111.2 ng/mL), and lactate dehydrogenase (LDH) [498 IU/dL] in the loculated group were significantly higher than those in the free-flowing group (TNF-alpha, 15.0 pg/mL; IL-1beta, 2.9 pg/mL; TGF-beta1, 6,117.3 pg/mL; PAI-1, 61.5 ng/mL, and LDH, 266 IU/dL). In both the loculated and free-flowing effusions, the levels of TGF-beta1 correlated positively with those of TNF-alpha (r = 0.51 and p < 0.001 vs r = 0.42 and p < 0.05, respectively) and IL-1beta (r = 0.52 and p < 0.001 vs r = 0.49 and p < 0.01, respectively), and the values of PAI-1 correlated positively with those of TNF-alpha (r = 0.59 and p < 0.001 vs r = 0.55 and p < 0.001, respectively), IL-1beta (r = 0.35 and p < 0.05 vs r = 0.47 and p < 0.01, respectively), and TGF-beta1 (r = 0.53 and p < 0.001 vs r = 0.58 and p < 0.001, respectively). In contrast, the levels of tPA correlated negatively with those of TNF-alpha (r = -0.37, p < 0.05) and IL-1beta (r =-0.56, p < 0.001) in loculated effusions. Twenty-seven of 42 patients with loculated effusions were classified into a secondary loculation group, which, compared with the primary loculation group, had significantly higher median levels of effusion TNF-alpha (119.2 vs 14.2 pg/mL, respectively; p = 0.001), IL-1beta (33.3 vs 3.4 pg/mL, respectively; p < 0.001), TGF-beta1 (13,152.7 vs 7746.0 pg/mL, respectively; p = 0.041), and PAI-1 (114.9 vs 94.1 pg/mL, respectively; p = 0.019). CONCLUSION: Compared with free-flowing effusions, fibrinolytic activity was depressed in loculated effusions. A higher intensity of pleural inflammation in loculated effusions may enhance the release of TNF-alpha, IL-1beta, and TGF-beta1, which may subsequently increase the levels of PAI-1. The imbalance of PAI-1 and tPA in pleural spaces may lead to fibrin deposition and loculation of pleural effusions.     

Expression of tissue factor and vascular endothelial growth factor is associated with angiogenesis in colorectal cancer

We examined the expression of tissue factor (TF) and vascular endothelial growth factor (VEGF) and the microvessel density (MVD) in 100 patients with colorectal cancer, and we investigated the relationship of the expression of TF or VEGF with angiogenesis. TF antigen was positive in 57.0% of all specimens. Incidence of TF expression was 41.2%, 45.5%, 52.6%, 84.6%, and 81.3% in tumors from patients in clinical stages I, II, IIIA, IIIB, and IV, respectively. TF expression was correlated with the Dukes' classification (P = 0.01) and the clinical stage of colorectal cancer (P = 0.02). VEGF antigen was positive in 64.0% of all specimens. Incidence of VEGF expression was 41.2%, 57.6%, 73.7%, 84.6%, and 75.0% in tumors from patients in clinical stages I, II, IIIA, IIIB, and IV, respectively. VEGF expression was correlated with the Dukes' classification (P = 0.01) but showed a weak association with the clinical stage (P = 0.08). MVD was significantly associated with the depth of invasion (P = 0.01), lymph node metastasis (P = 0.001), and liver metastasis (P = 0.02). The mean values of MVD were 7.5 +/- 2.8, 10.1 +/- 5.7, 14.6 +/- 5.8, 13.5 +/- 3.9, and 15.9 +/- 4.2 in tumors from patients in clinical stages I, II, IIIA, IIIB, and IV, respectively. A close relationship between VEGF and MVD (P < 0.001) and a significant correlation between TF expression and MVD were observed (P = 0.02). TF-positive carcinomas presented high MVD and VEGF expression (P < 0.001) more frequently than did TF-negative tumors. These results suggest that involvement of TF in the process of metastasis and progression of colorectal cancer may depend on increased angiogenesis.     

The influence of aetiology on inflammatory and neurohumoral activation in patients with severe heart failure: a prospective study comparing Chagas' heart disease and idiopathic dilated cardiomyopathy

Patients with Chagas' cardiomyopathy have the worst prognosis when compared to other aetiologies. It has been suggested that a more intense inflammatory activation could be responsible for this excessive mortality. We studied 35 patients with idiopathic dilated cardiomyopathy (IDC group) and 28 patients with Chagas' heart disease (Chagas' group) and 12 control subjects. We compared plasma tumor necrosis factor alpha (TNF-alpha), soluble TNF-alpha receptor type 1 (sTNF-R1), soluble Fas (sFas), interleukin 6 (IL-6), and brain natriuretic peptide type B (BNP) concentrations between the groups. TNF-alpha and IL-6 concentrations were higher in the IDC and Chagas groups as compared to controls (p<0.001 and p=0.001, respectively). sTNF-R1 concentration was higher in IDC after stratification for functional class (p=0.039), and there was a trend toward higher plasma TNF-alpha concentration in the Chagas' group (p=0.092). IL-6 concentration was higher in Chagas than in IDC (p=0.005). Higher IL-6 levels were associated with worse outcome (p=0.03 for Chagas; p=0.003 for IDC). sFas concentration was similar among groups. BNP concentrations were higher in IDC (350 pg/ml) and in Chagas (444.6 pg/ml) as compared to the controls (20.3 pg/ml; p<0.01). Higher BNP levels were associated with death and heart transplantation in both aetiologies. Inflammatory activation in Chagas heart disease differs from IDC and is associated with heart failure severity.     

Nutritional status, pulmonary functions, and exercise performance in COPD cases

The aim of this study was evaluation of nutritional status in attack and stable chronic obstructive pulmonary disease (COPD) cases, and investigating the relation between nutrition parameters and pulmonary functions, exercise performance and general healthy status. Anthropometric measures of 10 cases with attack (group 1), 14 stabilized COPD cases (group 2) and 10 control cases (group 3) were evaluated by measuring serum albumin, transferrin, tumor necrosis factor-alpha (TNF-alpha) levels, and fat-free mass (FFM) levels. Six minutes walking test (mwt) and SGRQ questionnaire were performed. Body mass index (BMI) of group 1 and 2 were significantly lower than group 3 (p= 0.023, p= 0.008, respectively). Albumin levels were significantly lower in group 1 than group 2 (p< 0.001) and 3 (p= 0.001). Levels of transferrin were significantly lower in group 1 compared with group 3 (p= 0.015). TNF-alpha levels were significantly high in group 1 compared with group 2 (p= 0.002), and high in group 2 compared with group 3 (p= 0.001). FFM was significantly low in group 1 compared with group 2 (p= 0.003), and low in group 2 compared with group 3 (p< 0.001). Mean distance of 6 mwt was 225.80 +/- 40.35 m in group 1, it was 405.16 +/- 95.51 m in group 2 (p< 0.001). A positive relation was seen between FFM and BMI, 6 mwt (p= 0.006, r= 0.481 ve p< 0.001, r= 0.830, respectively), between albumin levels and 6 mwt (p< 0.001, r= 0.850). A negative correlation was observed between TNF-alpha and 6 mwt (p< 0.001, r= -0.745). There was no statistical difference in antropometric measures between groups. Total score and daily life were statistically high in group 1 than group 2 (p= 0.009, p= 0.013). Although no changes was seen in antropometric measures of COPD cases, a nutritional defect was seen and a significant relation was observed between nutrition parameters and effort capacity.     

Coexpression of vascular endothelial growth factor and p53 protein in squamous cell carcinoma of the esophagus

OBJECTIVE: p53 plays a role in tumor angiogenesis, and vascular endothelial growth factor (VEGF) plays a key role in tumor angiogenesis. The aim of the present study was to clarify how expression of p53 protein participates in angiogenesis, and whether the coexpression of VEGF and p53 protein has a significance for angiogenesis and the clinicopathological features in esophageal squamous cell carcinoma (SCC). METHODS: Tissues samples were taken from 60 patients with esophageal SCC after surgery. The expression of VEGF and p53 protein in these SCC was examined immunohistochemically. Microvessel density (MVD) was determined by counting microvessels in tumor sections stained for Factor VIII-related antigen. Ki-67 labeling index (LI) was calculated, based on Ki-67 antigen immunostaining, as a proliferative marker. Apoptotic index (AI) was calculated, based on the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick end labeling, to evaluate apoptosis. RESULTS: VEGF expression was observed in 58.3%, and p53 protein expression was observed in 61.7% of the 60 patients. VEGF and p53 protein were significantly coexpressed in 26 (43.4%). Histological venous invasion (p < 0.01) and distant metastasis (p < 0.05) were significantly correlated with p53 protein expression. The two parameters were more frequently observed in the SCC with VEGF/p53 coexpression than in those without the coexpression. The MVD and Ki-67 LI were significantly higher (p < 0.01 and p < 0.001), and the AI was significantly lower (p < 0.001) in the SCC with p53 protein expression than in the SCC without it. The MVD and Ki-67 LI were higher, and the AI was lower in the SCC with VEGF/p53 coexpression than in those without the coexpression. The 5-yr survival rate in patients with the coexpression was poorer than in the other patients. CONCLUSION: These results suggest that mutant p53 expression is associated with angiogenesis and distant metastasis in esophageal SCC, and that the coexpression of p53 and VEGF may play an important role in angiogenesis, and have important clinical significance.     

Contribution of selected serum inflammatory mediators to the progression of chronic rheumatic valve disease, subsequent valve calcification and NYHA functional class

BACKGROUND AND AIM OF THE STUDY: The mechanism of the underlying principle of the progression of chronic rheumatic valve disease (RVD) and subsequent valve calcification are yet not clearly understood. The study aim was to determine whether serum markers of inflammation impact on the severity of chronic RVD, subsequent valve calcification and NYHA functional class. METHODS: The study group comprised 92 patients (27 males, 65 females; mean age 40 +/- 14 years) with RVD; the control group included 50 age- and gender-matched subjects without echocardiographic signs of RVD. All patients underwent echocardiographic of rheumatic valve severity, valve calcification and NYHA functional class. Levels of cytokines (interleukin-6 (IL-6), interleukin-2 receptor (IL-2R), interleukin-8 (IL-8), tumor necrosis factor-alpha (TNF-alpha)) and serum inflammatory markers (fibrinogen, high-sensitive C-reactive protein (hs-CRP)) were measured in all subjects. RESULTS: Plasma levels of IL-6, IL-8, IL-2R, TNF-alpha and hs-CRP were significantly higher in patients with RVD than in controls (p < 0.001). Significant correlations were identified between mitral score and fibrinogen (p = 0.002), IL-6 (p = 0.007), TNF-alpha (p < 0.001) and hs-CRP levels (p < 0.001). Fibrinogen, hs-CRP, IL-6, TNF-alpha and IL-2R levels correlated with functional class severity, while IL-6 and TNF-a levels correlated strongly with valve calcification (p < 0.001). CONCLUSION: The chronic phase of RVD is associated with ongoing serum inflammatory mediators which correlate strongly with the severity of valve involvement, valve scarring, subsequent valve calcification and decreasing functional status. Future research in this area should focus on whether anti-inflammatory drugs might reduce progression, morbidity and mortality in patients with chronic RVD.     

Time-course of changes in the levels of interleukin 6 in acutely decompensated heart failure

BACKGROUND: Although cytokine elevation has been demonstrated in chronic heart failure, little attention has been focused on cytokine levels during the acute stage. We examined the changes of cytokine levels in patients with acutely decompensated heart failure to investigate their relationship with severity of heart failure. METHODS: Patients with acutely decompensated heart failure (73 patients; 72+/-2 years) were included. Blood samples were taken from the peripheral vein on admission before the start of drug therapy, at 12, 24, 48 and 72 h as well as 1, 2 and 4 weeks after admission. Control data were obtained from age-matched normal patients who had no cardiovascular disease. Serum IL-6, IL-1beta and TNF-alpha levels were measured using the ELISA method. RESULTS: Mean IL-6, IL-1beta and TNF-alpha levels on admission were significantly higher than those in the control patients (p<0.001). IL-6 peaked at 12 h and declined thereafter, whereas IL-1beta and TNF-alpha remained unchanged throughout the duration of the study. Peak IL-6 significantly correlated with pulmonary wedge pressure on admission (r=0.332, p=0.0041). % change of IL-6 levels between peak (12 h after admission) and 24 h was significantly correlated with that of pulmonary wedge pressure between peak (on admission) and 24 h (r=0.308, p=0.0081). Peak IL-6 in patients treated with mechanical ventilation on admission was significantly higher than that in patients who underwent no mechanical ventilation (p<0.05). CONCLUSIONS: IL-6 levels possibly reflect the severity of heart failure and thus may be useful for the evaluation of disease status in acutely decompensated heart failure.     

Prognostic significance of vascular endothelial growth factor expression and microvessel density in carcinoma of ampulla of Vater

BACKGROUND/AIMS: To investigate whether the expression of vascular endothelial growth factor (VEGF) and microvessel density (MVD) are of prognostic significance in ampullary carcinoma. METHODOLOGY: Twenty-two resected tumor specimens from patients with ampullary carcinoma were immunohistochemically stained for VEGF and CD34 (surrogate for vessels) by streptavidin-peroxidase method. RESULTS: Expression of VEGF in tumor tissue was found in 50% of patients. The mean MVD for entire group was 26.4 +/- 12.8. A significantly higher MVD was observed in the tumors with positive VEGF expression (35.0 +/- 9.6) compared with that of negative VEGF expression (17.7 +/- 9.3) (p<0.01). The expression of VEGF and MVD were closely related lymph node status and tumor TNM stage. The positive expression rate of VEGF and the average MVD in patients with lymph node metastases were 85.7% and 33.1 +/- 10.8 respectively, which were significantly higher than those in patients without lymph node metastases (33.3% and 22.8 +/- 11.8 respectively) (p<0.05). The positive expression rate of VEGF and the average MVD in patients with stage III and were 75% and 36.3 +/- 8.4 respectively, which were significantly higher than those in patients with stage I (25% and 18.4 +/- 10.1 respectively) or stage II (50% and 23.8 +/- 13.4 respectively) (p<0.05). The Kaplan-Meier survival curves showed that the 3-year survival rate for patients with positive VEGF expression or a high MVD (9.1% and 10% respectively) were lower than those in patients with negative VEGF expression or a low MVD (63.64% and 58.33% respectively) (p<0.05). CONCLUSIONS: VEGF is positively correlated with MVD in ampullary carcinoma. VEGF and angiogenesis may play an important role in lymph node metastasis and progression of ampullary carcinoma. VEGF and MVD appear to be important prognostic predictor in patients with ampullary carcinoma.     

Increased productivity of tumor necrosis factor-alpha in helper T cells in patients with systolic heart failure

BACKGROUND AND AIMS: Whereas increased circulating proinflammatory cytokines, such as tumor necrosis factor-alpha (TNF-alpha), play an important role in heart failure, where and how TNF-alpha production is upregulated remains largely unknown. We studied the productivity of TNF-alpha in peripheral lymphocytes and underlying mechanisms in patients with heart failure. METHODS: Symptomatic NYHA II-IV patients with chronic heart failure with systolic dysfunction (n=39, aged 74+/-11, ejection fraction [EF]<==50%) were compared with asymptomatic NYHA I patients (n=18, aged 72+/-10, EF>50%) and normal subjects (n=15, aged 67+/-11). Lymphocyte subsets (CD3, CD4, and CD8) and intracellular production of TNF-alpha in peripheral leukocytes were quantified by immunofluorescent flow cytometry, and relationships between these parameters and circulating proinflammatory cytokines were analyzed. RESULTS: Subpopulation of TNF-alpha-producing CD4 was larger in NYHA II-IV patients (23.7% [18.0-28.6]) than in normal subjects (17.1% [6.5-19.5], p<0.05) and was correlated with plasma TNF-alpha levels (r=0.26, p<0.05), EF (r=-0.26, p<0.05), CD4/CD8 ratios (r=0.42, p<0.001), and subpopulation of TNF-alpha-producing monocytes (r=0.47, p<0.0001). Plasma levels of soluble CD14 and interleukin-12 (IL-12) were significantly higher in NYHA II-IV patients than in normal subjects (1971 ng/mL [1740-2375] vs. 1607 ng/mL [1530-1930], p<0.01; and 121 pg/mL [62-230] vs. 62 pg/mL [54-99], p<0.05, respectively), and plasma IL-12 levels were correlated with plasma TNF-alpha levels (r=0.41, p<0.001). CONCLUSIONS: Increased productivity of TNF-alpha in helper T cells, associated with their dominance over cytotoxic T cells, may partially contribute to an increase in circulating TNF-alpha levels in heart failure.     

Assay of pleural fluid interleukin-6, tumour necrosis factor-alpha and interferon-gamma in the diagnosis and outcome correlation of tuberculous effusion

OBJECTIVE: To assess the usefulness of interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) in the diagnosis and prediction of outcome of pleural tuberculosis. PATIENTS AND METHODS: Pleural fluid from 32 TB and 34 non-TB patients was sent for assay of IL-6, TNF-alpha and IFN-gamma. Clinical parameters at presentation and residual pleural scarring at completion of treatment were assessed for pleural TB cases. RESULTS: The pleural fluid Levels of IL-6, TNF-alpha and IFN-gamma in TB patients were significantly higher than those with non-TB effusions (P values of <0.001, 0.018 and <0.001, respectively by independent t-test). Utility of these cytokines for diagnosis of pleural TB was evaluated using receiver operating characteristic (ROC) curve analysis. The cut-off values for IL-6, TNF-alpha and IFN-gamma determined in this analysis were 4000, 4 and 60 pg/ml respectively, and their sensitivity and specificity were 90.6% and 76.5%, 90.6% and 79.4%, 100% and 100%, respectively. The pretreatment pleural fluid IL-6 levels had a positive correlation with the number of febrile days after treatment (Pearson correlation test: r=0.60, P=0.009). A negative correlation was found between the percentage reduction in pleural fluid cytokines after 2 weeks treatment and the extent of residual pleural scarring (IL-6: r=-0.62, P=0.041; TNF-alpha: r=-0.65, P=0.030; IFN-gamma: r=0.83, P=0.002). CONCLUSION: Pleural fluid IL-6, TNF-alpha and IFN-gamma assays are useful in the diagnosis of pleural TB. The initial IL-6 level correlates with the number of febrile days. The percentage change of cytokines after 2 weeks of treatment also helps to predict residual pleural scarring.     

Correlative studies on uPA mRNA and uPAR mRNA expression with vascular endothelial growth factor, microvessel density, progression and survival time of patients with gastric cancer

AIM: TO investigate the correlations between the expression of urokinase-type plasminogen activator (uPA) mRNA, uPA receptor (uPAR) mRNA and vascular endothelial growth factor (VEGF) protein and clinicopathologic features, microvessel density (MVD) and survival time. METHODS: In situ hybridization and immuno-histochemistry techniques were used to study the expressions of uPA mRNA, uPAR mRNA, VEGF and CD34 protein in 105 gastric carcinoma specimens. RESULTS: Expressions of uPA mRNA, uPAR mRNA and VEGF protein were observed in 61 (58.1%) cases, 70 (66.7%) cases and 67 (63.8%) cases, respectively. The uPA mRNA and uPAR mRNA positive expression rates in infiltrating-type cases (73.7%, 75.4%), stageⅢ-Ⅳ(72.1%, 75.4%), vessel invasion (63.2%, 69.9%), lymphatic metastasis (67.1%, 74.4%) and distant metastasis (88.1%, 85.7%) were significantly higher than those of the expanding-type (X2= 15.57, P= 0.001; X2=6.91, P=0.046), stageⅠ-Ⅱ(X2 = 19.22, P = 0.001; X2= 16.75, P= 0.001), non-vessel invasion (X2 = 11.92, P = 0.006; X2 = 14.15, P = 0.002), non-lymphatic metastasis (X2 = 28.41, P = 0.001; X2= 22.5, P=0.005) and non-distant metastasis (X2 = 12.32, P= 0.004; X2= 17.42, P = 0.002; X2 = 11.25, P = 0.012; X2 = 18.12, P = 0.002).The VEGF positive expression rates in infiltrating-type cases (75.4%), stageⅢ-Ⅳ(88.5%), vessel invasion (82.9%), lymphatic metastasis (84.3%) and distant metastasis (95.2%) were significantly higher than those of the expanding-type (X2 = 9.61, P = 0.021), stage I-II (X2=16.66, P = 0.001), non-vessel invasion (X2= 29.38, P = 0.001), non-lymphatic metastasis (X2 = 18.68, P = 0.005), and non-distant metastasis (X2= 22.72, P = 0.007; X2 = 21.62, P = 0.004). The mean MVD in the specimens positive for the uPA mRNA, uPAR mRNA and VEGF protein was markedly higher than those with negative expression groups. Moreover, a positive relation between MVD and uPA mRNA (rs = 0.199, P = 0.042), uPAR mRNA (rs = 0.278, P = 0.035), and VEGF (rs = 0.398, P = 0.048) expressions was observed. The mean survival time in cases with positive uPA mRNA, uPAR mRNA and VEGF protein expression or MVD value≥54.9 was significantly shorter than those in cases with negative expression or MVD value < 54.9. CONCLUSION: uPA and uPAR expressions are correlated with enhanced VEGF-induced tumor angiogenesis and may play a role in invasion and nodal metastasis of gastric carcinoma, thereby serving as prognostic markers of gastric cancer.     

Absence of Clinical Prognostic Value of Vascular Endothelial Growth Factor and Microvessel Density in Multiple Myeloma

Vascular endothelial growth factor (VEGF) is considered a potent stimulator of angiogenesis. In multiple myeloma (MM), it has been reported that bone marrow angiogenesis parallels tumor progression and correlates with a poor prognosis. To investigate the role of angiogenesis in MM, we investigated VEGF expression and microvessel density (MVD) in the bone marrow of 75 MM patients by immunohistochemical methods. VEGF expression was observed in 87.3% (62 of 71) of patients. MVD was 69.42 +/- 9.67 (mean +/- SE) compared with the normal control values of 26.81 +/- 2.85. MVD values were 73.98 +/- 11.27 and 36.04 +/- 6.99 in the VEGF-positive and VEGF-negative groups, respectively. The MVD of patients in the VEGF-positive group was significantly higher than in the VEGF-negative group (P = .045). However, there were no significant differences in various clinical parameters, such as age, sex, hemoglobin, platelet count, serum levels of albumin, calcium, creatinine, and beta2-microglobulin, and bone marrow plasma cell percentage, between the VEGF-positive and VEGF-negative groups. Multivariate analysis revealed that age, hemoglobin, platelet count, serum levels of albumin and creatinine, and bone marrow plasma cell percentage were correlated with overall survival, whereas VEGF expression or MVD was not. In conclusion, our results suggest that VEGF is highly expressed and that MVD is increased in MM, indicating that angiogenesis may play a role in MM. Although MVD in the bone marrow of the VEGF-positive group is significantly higher compared with the VEGF-negative group (P = .045), VEGF is not correlated with overall survival. Further studies that include other angiogenic factors are needed to determine the functional role of angiogenesis in MM.     

Relationship between the expression of iNOS,VEGF,tumor angiogenesis and gastric cancer

AIM：To in vestigate the relationship between the expression of inducible nitric oxide synthase(iNOS),vascular endothelial growth factor(VEGF),the microvascular density(MVD)and the pathological features and clinical staging of gastric cancer.METHODS:Immunohistochemical staining was used for detecting the expression of iNOS and VEGFin46resected specimens of gastric carcinoma;the monoclonal antibody against CD34 was used for displaying vascular endothelial cells,and MVD was detected by counting of CD34-positive vascular endothelial cells.RESULTS:Of 46resected specimens of gastric carcinoma,the rates of expressions of iNOS and VEGF were 58.70%and76.09%,respectively,and MVDaveraged55.59&#177;19.39,Judged by the standard TNM criteria,the rate of expression of iNOS in stageⅣ（84.46%)was higher than those in stageⅠ,Ⅱ,Ⅲ（Fish exact probabilities test,P=0.019,0.023and 0.033,respectively);the rates of expression of VEGFin stage Ⅲ,Ⅳ（76.0%,92.31%,respectively)were higher than those in stageⅠ,Ⅱ（Fis exact probabilities test,P=0.031,0.017,0.022and0.019).MVDs in stageⅢ,Ⅳ（64.72&#177;14.96,67.09&#177;18.29,respectively)were higher than those in stageⅠ,Ⅱ(t\2.378,4.015,2.503and2.450,P&lt;0.05,P&lt;0.001,P&lt;0.001,P&lt;0.05,respectively),In37gastric carcinoma specimens with lymph node metastasis,MVD(68.69&#177;18.07)and the rates of expression of iNOS and VEGF(70.27%,83.78%,respectively)were higher than those in the specimens with absence of metastasis(t=2.205,X^2=6.3587,X^2=6.2584,P&lt;0.01,P&lt;0.05,P&lt;0.05,respectively),MVD and the expressions of iNOS and EGF were not correlated to the location,size or grade of tumor,nor with the depth of invasion of tumor;MVDs in the positive iNOS and VEGF specimens(59.88&#177;18.02,58.39&#177;17.73,repectively)were higher than those in the negative iNOS and VEGF specimens(X^2=6.3587and 6.1574,P&lt;0.05,P&lt;0.05,respectively);thus the expressions of iNOS and VEGF was correlated to MVD,but the expression of iNOS was not correlated to that of VEGF,In addition.of the 46 surviving patients,the 5-year survival rate of patients with positive iNOS or VEGF tumors was significantly less than that of patients with negative iNOS-or VEGF tumors(X^2=4.3842and 5.4073,P&lt;0.05,P&lt;0.05.respectively).CONCLUSION:The expressions of iNOS and VEGF are colosely related to tumor angiogenesis,and are involved in the advancement and the lymph node metastasis;thusMVD and the expressions of iNOS and EGF may serve indexes for evaluating staging of gastric carcinoma and forecasting its risk of metastasis,which will help establish a comprehensive therapeutical measure of post-operative patients and provide a new approach to tumor therapy.     

Multiple myeloma with deletion of chromosome 13q is characterized by increased bone marrow neovascularization

Anti-angiogenesis therapy with thalidomide has been reported to have marked activity in multiple myeloma (MM). As cytogenetics is an independent prognostic factor in MM, we analysed bone marrow (BM) angiogenesis and cytogenetic abnormalities in 34 patients with active MM. BM microvessel density (MVD), as determined by staining with anti-CD34, was significantly higher in MM (MVD: 221 +/- 94 per mm2) than in controls (80 +/- 36; P < 0.0001). In patients with the presence of at least one unfavourable cytogenetic abnormality (deletion of 13q14, deletion of 17p13, aberrations of 11q), a significantly increased BM MVD was observed (254 +/- 93 vs. 160 +/- 60 in patients with absence of these abnormalities; P = 0.0035). Further analyses indicated that increased BM MVD was significantly correlated with deletion of 13q14 (259 +/- 96 vs. 188 +/- 80; P = 0. 026), but not with other cytogenetic, clinical and laboratory MM parameters. We conclude that BM neovascularization is particularly high in MM with deletion of 13q14, which provides a rationale for use of anti-angiogenic strategies in the treatment of MM with high-risk cytogenetics.     

Circulating cytokine levels compared to pain in patients with fibromyalgia -- a prospective longitudinal study over 6 months

OBJECTIVE: This prospective study examined circulating cytokines in patients with fibromyalgia (FM) over 6 months rather than at only one timepoint, and investigated correlations between serum cytokine concentrations and pain intensity in FM patients receiving multidisciplinary pain therapy. METHODS: Serum concentrations of proinflammatory cytokines interleukin 6 (IL-6), IL-8, and tumor necrosis factor-alpha (TNF-alpha) and antiinflammatory cytokines IL-4 and IL-10 were measured (Bio-Plex system) in 20 FM patients and 80 healthy subjects on admission and 10, 21, and 180 days after initiation of treatment and correlated to pain intensity. RESULTS: On admission, serum levels of IL-8 (p < 0.001) and TNF-alpha (p < 0.001), but not IL-6, were elevated in patients with FM. No significant difference in IL-4 and IL-10 was found between FM patients and controls. High IL-8 levels remained consistent during the followup, but TNF-alpha was already reduced after 10 days and until 6 months after therapy. After 6 months' treatment with multidisciplinary pain therapy, IL-8 and TNF-alpha levels were significantly lower than at the beginning (p < 0.05 for IL-8, p < 0.001 for TNF-alpha). IL-8 but not TNF-alpha serum levels were correlated with pain intensity (r = -0.782, p = 0.001) in FM patients after 6 months' multidisciplinary pain therapy. CONCLUSION: Our results suggest that proinflammatory cytokines TNF-alpha and IL-8 are involved in FM, but they do not apparently provoke the pain of FM directly. Multidisciplinary pain therapy modified the cytokine profile in patients with FM during the observation period.