Low levels of activated factor VII in systemic sclerosis

Introduction. Recent investigations show that activated factor VII, the primary enzyme in the extrinsic pathway of blood coagulation, exerts additional extra-coagulant functions, such as apoptosis and angiogenesis. On the basis of these recent acquisitions, the present study was aimed to evaluate activated factor VII in patients with systemic sclerosis and to establish a potential association with pathogenesis and complications of this severe autoimmune disorder. Materials and methods. Activated factor VII level was measured in twenty-eight consecutive scleroderma patients (2 men and 26 women, mean age 49.7 ± 14.8 years). The main clinical correlates of disease, such as disease activity, renal function, skin, vascular and lung involvement, were evaluated by clinical and instrumental investigations. Activated factor VII level was also evaluated in 28 sex and age matched controls. Results. Systemic sclerosis patients exhibited plasma activated factor VII activities significantly lower than those of healthy matched controls (15.2 versus 37.7 U/l, respectively; p < 0.001). No correlation was observed between plasma activated factor VII concentration and age, disease duration, disease subset, disease activity, renal, lung, skin and microvascular involvement. Conclusions. Results of our investigation provide first evidence of low activated factor VII activity in patients with systemic sclerosis. Reduced activated factor VII activity might be involved in the pathogenesis of the ischemic complications, by modulating apoptotic and angiogenetic processes. Contributed equally to this work.     

Elevation of serum lymphotactin levels in patients with systemic sclerosis

OBJECTIVE: To determine serum concentrations of lymphotactin, a Th1 chemokine, and their clinical association in patients with systemic sclerosis (SSc). METHODS: Lymphotactin levels were examined in serum samples from patients with SSc (n = 68), systemic lupus erythematosus (SLE; n = 42), or dermatomyositis (DM; n = 29), and healthy controls (n = 18) by enzyme linked immunosorbent assay. RESULTS: Serum lymphotactin levels were significantly elevated in SSc patients compared to patients with SLE or DM as well as controls. Serum lymphotactin levels were similar in patients with limited cutaneous SSc and diffuse cutaneous SSc (dSSc). Clinical correlation of elevated lymphotactin levels was not detected in the total group of patients with SSc, while elevation of lymphotactin levels was significantly associated with higher percentage vital capacity and percentage diffusing capacity of carbon monoxide, lower lung severity grade and serum IgG levels, and less frequent presence of short sublingual frenulum in patients with dSSc. CONCLUSION: Our results indicate that elevated serum lymphotactin levels correlate with relatively milder manifestations in dSSc, especially lower severity of lung involvement, suggesting that lymphotactin may play a role in the development of dSSc.     

Low serum levels of free and total insulin-like growth factor I (IGF-I) in patients with anorexia nervosa are not associated with increased IGF-binding protein-3 proteolysis

Patients with anorexia nervosa (AN) are GH resistant, with elevated GH levels and low serum levels of total insulin-like growth factor I (IGF-I). IGF-I action is modulated by IGF-binding proteins (IGFBPs), and a variety of catabolic states has been characterized by the presence of increased IGFBP-3 proteolysis. The present study was performed to examine the levels of free IGFs in AN and to clarify whether AN is associated with increased IGFBP-3 proteolytic activity. In 24 patients and 10 age-matched controls, the fasting serum concentrations of free IGF-I and -II were measured using ultrafiltration by centrifugation. In addition, GH, GH-binding protein, total IGFs, IGFBP-1 to -4, and IGFBP-3 proteolytic activity were measured. The IGFBPs were measured by both immunoassays and Western ligand blotting. Twelve of the patients were restudied 3 months after a minor increase in body mass index. In AN, the levels of GH-binding protein, free and total IGF-I, free IGF-II, and IGFBP-3 were significantly reduced; total IGF-II, IGFBP-2, and IGFBP-4 levels were unchanged; and IGFBP-1 was increased. No increased IGFBP-3 proteolytic activity could be detected in AN. In conclusion, the mechanisms responsible for the adaption of the GH-IGF-IGFBP axis in AN may be different from other catabolic conditions, because the low levels of free and total IGF-I in AN are not associated with increased IGFBP-3 proteolysis.     

非酒精性脂肪性肝炎患者血清成纤维细胞生长因子-21和脂联素水平变化及其临床意义探讨

目的 探讨非酒精性脂肪性肝炎(NASH)患者血清成纤维细胞生长因子-21(FGF21)、脂联素(APN)和胰岛素抵抗水平变化及其临床意义。方法 2 018年7月～2019年8月我院就诊的41例NASH患者和41例同期健康人群,检测空腹血糖(FBG)和空腹胰岛素(FINS),计算稳态模型胰岛素抵抗(HOMA-IR)指数,采用ELISA法检测血清FGF21水平,采用放射免疫法检测血清APN水平。结果 NASH患者FBG水平为(8.7±1.6)mmol/L,显著高于健康人【(5.6±0.5)mmol/L,P<0.05】,FINS水平为(30.9±9.8)μIU/mL,显著高于健康人【(12.5±3.6)μIU/mL,P<0.05】,HOMA-IR水平为(1.9±1.1),显著低于健康人【(3.8±2.5),P<0.05】;NASH血清FGF21水平为(279.7±106.4)pg/mL,显著高于健康人【(141.2±62.5) pg/mL,P<0.05】,血清APN水平为(2.9±0.8)μg/mL,显著低于健康人【(4.1±1.0)μg/mL,P<0.05】;10例重度组患者血清FGF21水平为(285.7±116.4)pg/mL,显著高于16例中度组和15例轻度组【分别为(174.5±75.3)pg/mL和(150.7±65.5)pg/mL,P<0.05】,血清APN水平为(2.1±0.5)μg/mL,显著低于中度组和轻度组【分别为(2.6±0.9)μg/mL和(3.0±1.1)μg/mL,P<0.05】,FBG水平为(9.9±2.6)pg/mL,显著高于中度组和轻度组【分别为(7.5±2.1)pg/mL和(6.6±1.5)pg/mL,P<0.05】,FINS水平为(35.9±8.8)pg/mL,显著高于中度组和轻度组【(27.1±4.0)pg/mL和(15.5±3.6)pg/mL,P<0.05】,HOMA-IR水平为(1.5±1.0),显著低于中度组和轻度组【分别为(1.7±1.2)和(2.1±1.0),P<0.05】;17例存在高血压糖尿病或高脂血症的NASH患者血清FGF21水平为(315.7±99.2)pg/mL,显著高于24例无合并症组【(254.2±91.1)pg/mL, P<0.05】,血清APN水平为(2.6±0.7)μg/mL,显著低于无合并症组【(3.1±0.8)μg/mL,P<0.05】。结论 合并有高血压、糖尿病和高脂血症的NASH患者血清FGF21水平升高,而血清APN和胰岛素抵抗指标降低,检测NASH患者血清FGF21、APN和胰岛素抵抗水平,有助于评估患者病情。     

Increased serum soluble CD40 levels in patients with systemic sclerosis

OBJECTIVE: To determine serum levels of soluble CD40 (sCD40) and clinical association in patients with systemic sclerosis (SSc). METHODS: Serum sCD40 levels were examined by ELISA in 49 patients with SSc, 15 patients with systemic lupus erythematosus, and 26 healthy individuals. sCD40 levels in plasma samples, which were obtained at the same time, were also determined. SSc patients were grouped into 22 patients with limited cutaneous SSc (lcSSc) and 27 patients with diffuse cutaneous SSc (dcSSc). RESULTS: There was no significant difference between sCD40 levels of sera and those of plasma. Serum sCD40 levels were significantly elevated in patients with SSc compared to patients with systemic lupus erythematosus and controls (p < 0.001). Serum sCD40 levels were higher in patients with lcSSc than in those with dcSSc (p <0.001). There was no correlation between sCD40 and sCD40 ligand levels in patients with SSc. CONCLUSION: Elevated serum sCD40 levels were associated with lcSSc. These results suggest that the blockade of CD40/CD40 ligand interaction could be a potential therapeutic strategy in SSc.     

Plasma plasmin-alpha2-plasmin inhibitor complex levels are increased in systemic sclerosis patients with pulmonary hypertension

OBJECTIVE: To determine the frequency and clinical significance of plasma plasmin-alpha(2)-plasmin inhibitor complex (PIC) in patients with systemic sclerosis (SSc). METHODS: Plasma samples from 74 patients with SSc and 32 healthy volunteers were examined by a specific enzyme-linked immunosorbent assay. RESULTS: Elevated plasma PIC levels were present in 35 of the 74 patients (47.3%) with SSc. The patients with elevated plasma PIC levels had pulmonary hypertension (PH) at a significantly higher incidence than those with normal PIC levels (31.4 vs 7.7%, P<0.01). When PH was classified into isolated PH (IPH) and secondary PH (SPH), the presence of IPH was significantly greater in patients with elevated PIC levels than in those with normal levels (25.7 vs 5.1%, P<0.02). CONCLUSIONS: These results suggest that plasma PIC levels may be a marker of PH, especially IPH, in patients with SSc.     

Clinical features of patients with systemic sclerosis accompanied by rheumatoid arthritis

OBJECTIVE: To determine the incidence of patients with both systemic sclerosis and rheumatoid arthritis (SSc-RA) and the clinical features of those with SSc-RA. METHODS: All 173 patients with systemic sclerosis in our clinic were investigated. RESULTS: Of the 173 patients with systemic sclerosis, 9 (5.2%) developed rheumatoid arthritis (RA). At the first visit, arthritis prior to Raynaud's phenomenon, increased C-reactive protein (CRP), and elevated rheumatoid factor (RF) were seen in patients with SSc-RA at a significantly higher incidence than in those without (44.4% versus 4.8%, p < 0.01; 55.6% versus 13.6%, p < 0.001; 247.2 +/- 312.1 versus 47.9 +/- 54.3 IU/ml, p < 0.001, respectively). Furthermore, in 8 of the 9 patients with SSc-RA, CRP was increased before the diagnosis of RA. CONCLUSION: These results suggest that systemic sclerosis patients with elevated RF and a history of arthralgia prior to Raynaud's phenomenon should be followed up with serial measurements of CRP due to their risk of developing RA.     

Endothelin-1 serum levels correlate with MCP-1 but not with homocysteine plasma concentration in patients with systemic sclerosis

OBJECTIVES: To determine whether homocysteine (Hcy) plasma levels are correlated with molecules indicative of endothelial cell and fibroblast activation, including endothelin-1 (ET-1) and monocyte chemoattractant protein-1 and -3 (MCP-1, MCP-3), in patients with systemic sclerosis (SSc). METHODS: Eighty-two patients were enrolled in this study; the control group included 75 age- and sex-matched subjects. Plasma Hcy was determined by high-performance liquid chromatography; folic acid, and vitamin B(12) plasma levels were determined by a chemiluminescence method. ET-1, MCP-1, and MCP-3 were determined by enzyme-linked immunosorbent assay (ELISA). Analysis of the 677C-->T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene was performed by polymerase chain reaction (PCR) and digestion with the enzyme HinfI. RESULTS: Hcy levels were lower in patients whereas ET-1 was significantly higher in patients and correlated with MCP-1. Stratification of the patients on the basis of Hcy levels was not associated with any statistical difference in the concentration of ET-1, MCP-1, and MCP-3. Patients with diffuse disease presented the highest levels of ET-1 and MCP-1. The distribution of the MTHFR genotypes was not different in patients and controls. CONCLUSIONS: In SSc, Hcy plasma concentration does not influence ET-1, MCP-1, or MCP-3 levels. On the contrary, ET-1, a marker of vascular activation, correlates with MCP-1, a chemokine involved in the fibrotic process of SSc.     

Elevated serum levels of soluble CD146 in patients with systemic sclerosis

CD146, a transmembrane glycoprotein member of the immunoglobulin superfamily, acts as an adhesion molecule that helps maintain the cell monolayer. Human endothelial cells expressing CD146 are involved in angiogenesis and inflammation. Recently, we developed a sandwich ELISA for detecting soluble CD146 (sCD146) in human serum specimens. The aim of this study is to determine serum levels of sCD146 in patients with systemic sclerosis (SSc) and to examine the relationship between sCD146 levels and clinical manifestations. We quantified serum sCD146 levels in 47 serum samples from patients fulfilling criteria for SSc, 23 serum samples from patients fulfilling criteria for rheumatoid arthritis (RA), and 25 healthy controls. We also investigated the relationship between sCD146 levels and various clinical characteristics with SSc patients. Levels of sCD146 were significantly higher in the 47 patients with SSc than in the 25 healthy controls and 23 patients with RA (12.50 vs. 6.91 vs. 9.95 ng/ml; p < 0.001). Serum sCD146 levels in SSc patients with pulmonary arterial hypertension (PAH) were lower than in SSc patients without PAH (10.12 vs.13.17 ng/ml; p < 0.01). The serum levels of sCD146 were elevated in patients with SSc. However, decreased sCD146 levels were observed in SSc patients with PAH. Further studies are necessary to elucidate the sources and the mechanisms.     

High levels of serum basic fibroblast growth factor in children with biliary atresia

BACKGROUND/AIMS: The purpose of this study was to investigate the relationship between basic fibroblast growth factor (bFGF) and clinical outcome in biliary atresia (BA) patients after surgical treatment. METHODOLOGY: Sixty-five pediatric patients with BA post Kasai operation and 12 healthy children were recruited. The patients were categorized into 2 groups according to their serum levels of total bilirubin (TB < 2, no jaundice vs. TB > or = 2mg/dL, persistent jaundice) and alanine aminotransferase (ALT < 100 vs. ALT > or = 100 U/L). The serum bFGF levels were determined by enzyme-linked immunosorbent assay. RESULTS: The mean serum bFGF level of the BA patients was higher than that of normal controls (14.2 +/- 1.6 vs. 9.0 +/- 2.2 pg/mL, p < 0.03). Serum bFGF levels in the persistent jaundice group were significantly higher than those without jaundice (17.6 +/- 3.1 vs. 11.9 +/- 1.5 pg/mL, p < 0.04). Additionally, patients with serum ALT > or = 100 U/L had higher serum levels of bFGF than those with serum ALT < 100 U/L (19.0 +/- 2.4 vs. 8.9 +/- 1.6 pg/mL, p < 0.0005). In the no jaundice group, serum bFGF levels were significantly elevated in the patients with portal hypertension (PH) compared with those without PH (15.8 +/- 2.2 vs. 8.6 +/- 1.9 pg/mL, p < 0.02). CONCLUSIONS: The significant elevation in bFGF levels is associated with the presence of PH and the severity of hepatic damage. bFGF may play a role in the pathogenesis of progressive inflammation and the perpetuation of PH in BA patients after Kasai procedure.     

Selective recognition of fibroblast growth factor-2 by the long pentraxin PTX3 inhibits angiogenesis

The long pentraxin PTX3 is a soluble pattern recognition receptor produced by monocytes and endothelial cells that plays a nonredundant role in inflammation. Several pathologic conditions are characterized by local production of both PTX3 and the angiogenic fibroblast growth factor-2 (FGF2). Here, solid-phase binding assays demonstrated that PTX3 binds with high affinity to FGF2 but not to a panel of cytokines and growth factors, including FGF1, FGF4, and FGF8. Accordingly, PTX3 prevented (125)I-FGF2 binding to endothelial cell receptors, leading to specific inhibition of FGF2-induced proliferation. PTX3 hampered also the motogenic activity exerted by endogenous FGF2 on a wounded endothelial cell monolayer. Moreover, PTX3 cDNA transduction in FGF2-transformed endothelial cells inhibited their autocrine FGF2-dependent proliferation and morphogenesis in vitro and their capacity to generate vascular lesions when injected in nude mice. Finally, PTX3 suppressed neovascularization triggered by FGF2 in the chick embryo chorioallantoic membrane with no effect on physiologic angiogenesis. In contrast, the short pentraxin C-reactive protein was a poor FGF2 ligand/antagonist. These results establish the selective binding of a member of the pentraxin superfamily to a growth factor. PTX3/FGF2 interaction may modulate angiogenesis in various physiopathologic conditions driven by inflammation, innate immunity, and/or neoplastic transformation.     

Dehydroepiandrosterone sulphate serum levels in systemic sclerosis

OBJECTIVE: To evaluate in a cohort of women with systemic sclerosis (SSc) the dehydroepiandrosterone sulphate (DHEAS) serum levels and their relationship with disease severity. METHODS: DHEAS serum concentrations were measured by radioimmunoassay in 40 SSc patients and compared with those in 40 controls matched for sex and reproductive status. IL-2 sR alpha was evaluated as a disease activity index. A preliminary organ/system severity scale proposed by Medsger et al. in 1999 was used to evaluate disease severity. RESULTS: Mean serum levels of DHEAS in SSc women of childbearing age were significantly lower than in controls (0.87 +/- 0.85 microgram/ml versus 2.75 +/- 0.42 micrograms/ml; p < 0.001). On the contrary, no difference was found between postmenopausal women and controls. A reduction below the 95% confidence limits was found in 10 out of 11 patients of childbearing age and in 8 out of 29 postmenopausal women, respectively. In 5 out of 11 patients of childbearing age taking steroids for their SSc (< 10 mg/daily) DHEAS levels were significantly lower than in patients not taking steroids (p = 0.01). On the contrary, 16 out of 29 postmenopausal women using steroids had lower DHEAS concentrations than in patients not taking steroids, although the difference was not statistically significant. There was no statistically significant difference in DHEAS levels between patients with diffuse or limited SSc, or between those with or without organ system involvement. No correlations were found either in pre- and post-menopausal steroid nonusers, or in limited and diffuse subsets, between DHEAS levels and age, postmenopausal years, disease duration, IL-2 sR alpha, disease organ/system severity scale. CONCLUSION: Our data show that, as in other autoimmune diseases, low serum DHEAS is a feature of premenopausal SSc patients. More extensive prospective studies are needed to define the exact role of DHEAS dysregulation in SSc.     

Sevelamer hydrochloride and calcium bicarbonate reduce serum fibroblast growth factor 23 levels in dialysis patients

Fibroblast growth factor 23 (FGF23) is a member of the fibroblast growth factor superfamily which displays a strong phosphaturic action and an inhibition of vitamin D 1-alpha hydroxylase activity. Fourty-six patients undergoing maintenance hemodialysis therapy participated in the study. They were randomly divided into 2 groups, and treated with either 3 g sevelamer hydrochloride+3 g of calcium bicarbonate (CaCO3), or 3 g of CaCO3 alone. Serum FGF23 levels were determined by a sandwich enzyme-linked immunosorbent assay (ELISA) system that detects the intact form of FGF23 molecules. Although the serum inorganic phosphate (Pi) levels were comparable before treatment, the levels were significantly lower in the patients treated with sevelamer hydrochloride+CaCO3 than those with CaCO3 alone after 4 weeks of treatment (P<0.05). Serum FGF23 levels significantly decreased after 4 weeks of the treatment with sevelamer hydrochloride+CaCO3 from the pretreatment levels (P<0.05), while no changes were found in the patients treated with CaCO3 alone. Thus, the use of sevelamer hydrochloride and CaCO3 reduced serum FGF23 levels in dialysis patients presumably through inhibiting phosphate load into the intestine.     

Transforming growth factor beta1 gene polymorphism in patients with systemic sclerosis

OBJECTIVE: To determine whether transforming growth factor beta1 (TGFbeta1) gene DNA polymorphism is associated with pathogenesis in the fibrosis of patients with systemic sclerosis (SSc). METHODS: Eighty-seven Japanese patients with SSc including 30 with diffuse type and 57 with limited type together with 110 unrelated controls were investigated. Pulmonary fibrosis was determined in 34 SSc patients using high-resolution chest computed tomography. TGFbeta1 genetic polymorphisms were analyzed in 2 loci; T869C (Leu10Pro) in codon 10 at exon 1, and C-509T in the promoter region using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Neither the genotype of T/C polymorphism in T869C nor C/T polymorphism in C-509T revealed any difference in distribution between SSc and controls. In the group of SSc patients with pulmonary fibrosis, a weak but significantly high frequency (p = 0.05) of TC+CC (the presence of C allele) in T869C, and CT+TT (the presence of T allele) in C-509T was found. Compared with controls, the pulmonary fibrosis group showed no difference in the highly frequent alleles. CONCLUSION: Our results suggest that TGFbeta1 polymorphisms do not play a role in the pathogenesis of SSc, even though there remains the possibility of a risk factor for genetic susceptibility to pulmonary fibrosis.     

Serum basic fibroblast growth factor levels in patients with ischemic heart disease

Background: Being a potent promoter of endothelial and smooth muscle cell proliferation, basic fibroblast growth factor (bFGF) is presumed to play a key role in coronary collateral development and atherogenesis. Purpose: To characterize serum bFGF levels in patients with ischemic heart disease. Methods: The study population consisted of patients with angina (n=33) and after uncomplicated myocardial infarction (n=12). The number of significantly stenosed (≥50%) vessels and angiographic coronary collateral score were noted. Blood was drawn immediately prior to elective coronary angiography in study patients for bFGF levels. Twenty healthy, age-matched subjects served as control for serum bFGF. Results: Serum bFGF levels were undetectable in all 20 control subjects, but were detectable in 15/33 (45%) patients with angina and 3/12 (25%) post-infarction patients, respectively (P=0.002). Serum bFGF levels were detectable in 13/23 (57%) patients with 0- or 1-vessel disease, as compared with 5/22 (23%) patients with 2- or 3-vessel disease (P<0.05). Detectable serum bFGF levels were not in correlation with coronary collateral score (P=1). Conclusions: Serum levels of bFGF are elevated in patients with ischemic heart disease, particularly in those with minimal coronary artery disease. We postulate that detectable serum bFGF levels reflect active atherogenesis rather than myocardial collateral development.