低氧诱导因子-1α和内皮素-1基因在大鼠低氧性肺动脉高压中的表达

目的　探讨低氧诱导因子 1(HIF 1α)和内皮素 1(ET 1)基因表达在低氧性肺动脉高压发病过程中的变化和作用。方法　复制低氧性肺动脉高压大鼠模型 ,测定平均肺动脉压 ,弹力纤维染色显示腺泡内肺动脉 ,用放射免疫法测ET含量 ,原位杂交方法进行检测HIF 1αmRNA。结果　HIF 1αmRNA在低氧各组腺泡内肺动脉有表达 ,低氧 14d组 (0 2 5 6 9± 0 0 46 8)和低氧 2 8d组 (0 2 2 5 8±0 0 45 3)染色强于低氧 5d组 (0 145 5± 0 0 2 72 )和正常组 (0 110 9± 0 0 2 2 4) ;ET 1mRNA在低氧各组腺泡内肺动脉有表达 ,低氧 14d组 (0 412 2± 0 0 783)和低氧 2 8d组 (0 36 84± 0 0 72 9)染色强于低氧5d组 (0 2 0 17± 0 0 34 9)和正常组 (0 185 5± 0 0 36 1) ,HIF 1α和ET 1基因表达在H14d组和H2 8d组明显增加 (P <0 0 5 )。肺动脉血中ET 1含量在H14d组 [(15 8 78± 2 5 14)pg/ml]和H2 8d组 [(142 93± 2 3 38)pg/ml]明显高于H5d组 [(79 6 8± 12 5 4)pg/ml]和正常组 [(6 5 37± 10 82 )pg/ml](P <0 0 5 ) ;H14d组 [(34 0± 5 8)mmHg]和H2 8d组 [(2 9 0± 4 7)mmHg]的mPAP也明显高于H5d组[(19 0± 3 5 )mmHg]和正常组 [(17 0± 2 8)mmHg](P <0 .0 5 ) ,并且与肺动脉血中ET 1含量呈正比(rs=0     

Urinary levels of RANTES and M-CSF are predictors of lupus nephritis flare

Objective: To explore a new predictor of renal flares after successful inductive treatment for diffuse proliferative glomerulonephritis(DPGN) in patients with lupus nephritis. Methods: A cohort of patents with SLE DPGN who were treated initially with prednisone and cyclophosphamide were studied. Those who responded to inductive treatment were followed up for the occurrence of renal flares. Urinary levels of RANTES, MCP-1 and M-CSF were measured by ELISA. Other clinical and laboratory data were collected. The predictors and outcome of renal flare were analyzed. Results: Seventy-three qualified patients with SLE DPGN were investigated. After a mean follow-up of 24.5 ± 6.4 months, 22 patients experienced renal flares. The median time to relapse was 14.1 ± 4.1 months. The patients experiencing renal flare showed higher urinary RANTES, MCP-1 and M-CSF. However, independent predictors of renal flares were increased urinary RANTES and M-CSF. Eight patents developed doubling of the serum creatinine (CRX2) level. The occurrence of renal flares was the only predictor of CRX2. Conclusions: Persistently increased urinary levels of RANTES and M-CSF after initial remission are predictors of renal flare in patients with SLE DPGN. Our results indicate monitoring urinary pro-inflammatory factors may direct us in managing those patients. Received 6 September 2006; returned for revision 30 November 2006; returned for final revision 23 February 2007; accepted by G. Wallace 19 March 2007     

Important considerations in pregnant patients with lupus nephritis

Introduction: In the last few decades, identification of predictors of pregnancy outcome and appropriate pregnancy planning have significantly reduced the maternal and fetal risks in pregnant women with lupus nephritis.

Areas covered: Successful pregnancies have been reported even in women with chronic renal disease and renal insufficiency. However, refractory hypertension and severe renal or cardiac chronic dysfunction are still considered contraindications to pregnancy. Pre-term delivery and fetal growth restriction may still occur in SLE patients more frequently than in healthy women, even in pregnancies regularly planned and monitored by a team of nephrologists and gynaecologists.

Expert commentary: Stable disease remission is the most important factor for a successful pregnancy. In case of flare-ups of lupus, timely diagnosis and appropriate management may ensure a successful outcome in the majority of pregnant women. The negative role of anti-phospholipid antibodies and of chronic arterial hypertension may be countered with appropriate anticoagulation and anti-hypertensive therapy. Further studies are needed to better assess the possible impact of pregnancy on the long-term outcome of lupus nephritis.     

Association of miRNA-145 expression in vascular smooth muscle cells with vascular damages in patients with lupus nephritis

miRNAs have been found to contribute to the regulation of multiple cellular processes, including cell apoptosis, differentiation and proliferation. The patients with lupus nephritis (LN) exhibit thickened renal vascular membrane and highly proliferative vascular smooth muscle cells (VSMCs). Of various miRNAs discovered, miR-145 is essential to mediate the proliferation of VSMCs and the formation of atherosclerotic plaques. In this study, we studied the pathological and vascular damage of renal LN, and the correlation between miR-145 expression in VSMCs and the vascular damages. Serum, urine, and renal biopsies were obtained from 41 patients with active LN. The serum and urinary VEGF levels were examined to confirm the renal damage of each patient. Biopsies were stained to observe the glomerular segmental lesions, sclerosis, and to evaluate the vascular damages. The expression of miR-145 was also examined to determine the correlation between its expression and the vascular damages. The expression of miR-145 was mainly detected in the renal VSMCs and the epithelial cells of glomerular proximal convoluted tubule. Nevertheless, the expression of miR-145 reduced as the tunicae media vasorum ratios increased, indicating the development of LN inhibits the expression of miR-145. Furthermore, our studies revealed no significant correlation among renal interstitial vascular damage, glomerular damage and severity classification of LN. Therefore, we suggest the damage of renal interstitial vascular should be considered as one of the factors to evaluate the severity of the LN.     

转染缺氧诱导因子-1α对肺癌细胞A549生长的影响

目的:研究缺氧诱导因子-1α(HIF-1α)对人肺癌细胞株A549体内、外生长的影响,并探讨其可能机制。方法:将HIF-1α转染人肺癌细胞A549中,观察A549细胞生长。建立人肺癌裸鼠模型,随机将其分为两组:对照组(A组,10只),HIF-1α转染细胞组(B组,10只)。1月后,切除瘤灶、称瘤重、计算促瘤率。标本用免疫组化测定增殖细胞核抗原(PCNA),同时用Westernblot检测HIF-1α、凋亡抑制蛋白survivin和bcl-2的表达。结果:HIF-1α转染A549细胞后,细胞生长速率加快。转染HIF-1α的裸鼠肿瘤生长较对照组快,A组、B组的瘤重分别为(2.51±0.33)g,(3.44±0.40)g,促瘤率为37%。免疫组化和Westernblot提示:B组的PCNA表达比A组高,B组HIF-1α、survivin、bcl-2的蛋白水平明显高于A组。结论:转染HIF-1α体内、外均可促进肺癌A549细胞的生长,其机制可能与其能促进细胞恶性增殖和抑制凋亡有关。     

Clinical and functional consequences of anti-properdin autoantibodies in patients with lupus nephritis

Properdin is the only positive regulator of the complement system. In this study, we characterize the prevalence, functional consequences and disease associations of autoantibodies against properdin in a cohort of patients with autoimmune disease systemic lupus erythematosus (SLE) suffering from lupus nephritis (LN). We detected autoantibodies against properdin in plasma of 22·5% of the LN patients (16 of 71) by enzyme-linked immunosorbent assay (ELISA). The binding of these autoantibodies to properdin was dose-dependent and was validated by surface plasmon resonance. Higher levels of anti-properdin were related to high levels of anti-dsDNA and anti-nuclear antibodies and low concentrations of C3 and C4 in patients, and also with histological signs of LN activity and chronicity. The high negative predictive value (NPV) of anti-properdin and anti-dsDNA combination suggested that patients who are negative for both anti-properdin and anti-dsDNA will not have severe nephritis. Immunoglobulin G from anti-properdin-positive patients’ plasma increased the C3b deposition on late apoptotic cells by flow cytometry. Nevertheless, these IgGs did not modify substantially the binding of properdin to C3b, the C3 convertase C3bBb and the pro-convertase C3bB, evaluated by surface plasmon resonance. In conclusion, anti-properdin autoantibodies exist in LN patients. They have weak but relevant functional consequences, which could have pathological significance.     

狼疮肾炎患者PBMC-CM对系膜细胞表达MCP-1的影响

目的探讨狼疮肾炎（心）患者外周血单个核细胞条件培养液（PBMC-CM）对系膜细胞表达单核细胞趋化蛋白-1（MCP-1）的影响。方法用免疫组化观察LN患者PBMC-CM对系膜细胞表达MCP-1和蛋白激酶C（PKC）的影响。结果LN患者PBMC-CM干预系膜细胞后,系膜细胞增多,培养上清Ⅳ型胶原和层黏蛋白表达增加,系膜细胞MCP-1、PKC表达增加,呈直线相关关系。结论LN患者PBMC-CM能刺激系膜细胞增殖,促进细胞外基质增多及MCP-1的表达增加,从而可能参与LN发病。     

Serum interleukin-17 levels are associated with nephritis in childhood-onset systemic lupus erythematosus

OBJECTIVES:

To determine the serum interleukin-17 (IL-17) levels in childhood-onset systemic lupus erythematosus patients and to evaluate the association between IL-17 and clinical manifestations, disease activity, laboratory findings and treatment.

METHODS:

We included 67 consecutive childhood-onset systemic lupus erythematosus patients [61 women; median age 18 years (range 11-31)], 55 first-degree relatives [50 women; median age 40 years (range 29-52)] and 47 age- and sex-matched healthy controls [42 women; median age 19 years (range 6-30)]. The childhood-onset systemic lupus erythematosus patients were assessed for clinical and laboratory systemic lupus erythematosus manifestations, disease activity [Systemic Lupus Erythematosus Disease Activity Index (SLEDAI)], cumulative damage [Systemic Lupus International Collaborating Clinics/American College of Rheumatology (ACR) Damage Index] and current drug use. Serum IL-17 levels were measured by an enzyme-linked immunosorbent assay using commercial kits.

RESULTS:

The median serum IL-17 level was 36.3 (range 17.36-105.92) pg/mL in childhood-onset systemic lupus erythematosus patients and 29.47 (15.16-62.17) pg/mL in healthy controls (p=0.009). We observed an association between serum IL-17 levels and active nephritis (p=0.01) and migraines (p=0.03). Serum IL-17 levels were not associated with disease activity (p=0.32), cumulative damage (p=0.34), or medication use (p=0.63).

CONCLUSION:

IL-17 is increased in childhood-onset systemic lupus erythematosus and may play a role in the pathogenesis of neuropsychiatric and renal manifestations. Longitudinal studies are necessary to determine the role of IL-17 in childhood-onset systemic lupus erythematosus.     

LFA-1与抗CD3 mAb共刺激对狼疮肾炎PBMC增殖和IgG合成的影响

目的 :探讨LFA 1与抗CD3mAb共刺激对狼疮肾炎 (LN)患者外周血单个核细胞 (PBMC)增殖与免疫球蛋白 (IgG)合成的影响。方法 :分离LN患者及健康人 (正常对照 )的PBMC ,采用Ficoll梯度密度离心法。细胞增殖实验采用3 H TdR掺入法。IgG测定采用ELISA法。结果 :在抗CD3mAb诱导下 ,2 9例LN非活动期和活动期PBMC中 ,3 H TdR的掺入量和IgG的合成均增加 ,且活动期的增殖强于非活动期 (P <0 .0 1) ;而单纯用抗CD3mAb对 12例正常对照的PBMC没有影响 (P >0 .0 5 )。抗CD3mAb与抗LFA 1mAb共刺激 ,均可增加LN患者 (活动期和非活动期 )及正常对照PBMC3 H TdR掺入量和IgG合成 ,其中活动期增殖效应强于非活动期 (P <0 .0 1) ,后者又强于正常对照 (P <0 .0 1)。抗LFA 1中和抗体的加入 ,可明显抑制LFA 1诱导的共刺激效应 (P <0 .0 1)。结论 :LFA 1促进LN患者PBMC增殖和IgG合成 ,可能是其参与LN发病的机制之一     

Glomerular necrotic lesions and long-term outcomes among patients with proliferative lupus nephritis

Objectives: Although necrotic lesions are common in proliferative lupus nephritis (LN), little is known about the impact of these lesions on outcomes. This study was undertaken to investigate the impact of glomerular necrotic lesions on renal outcomes of doubling serum creatinine in patients with class III and IV LN and necrotic lesions. Methods: 52 patients with ISN/RPS class III or IV LN were enrolled in this retrospective study, with mean follow-up of 7.4 years. All patients underwent a repeat biopsy at 12-18 months after a baseline biopsy. Results: The prevalence of necrotizing lesions was observed in 24% of those with class III versus 70.4% with class IV (P = 0.001). The rate of no remission was 44% and 22.2% in those with and without necrosis (P = 0.007), respectively. The doubling of serum creatinine was observed in 32% of those with necrosis and in 14.8% with no necrosis (P = 0.01). The chronicity index in the repeat biopsy was significantly worse among those with necrosis. Conclusions: Glomerular necrosis identifies lupus nephritis patients at the greatest risk for progression to renal failure. Proactive intervention and possibly more aggressive induction therapies in patients with necrotizing lesions may protect the kidneys from developing chronic renal impairment.     

血清抗C1q抗体在系统性红斑狼疮疾病活动性判断及狼疮肾炎诊断中的价值

目的探讨抗C1q抗体(C1qAb)在系统性红斑狼疮(SLE)活动性及狼疮肾炎(LN)诊断和疾病活动性判断中的价值。方法采用酶联免疫吸附法检测SLE患者(n=89)、疾病对照组(n=56)和正常对照组(n=42)血清中的抗C1q抗体阳性率,并与SLE患者临床实验室指标﹑活动性评分进行分析。结果 C1qAb的阳性率在SLE患者中显著高于疾病对照组和正常对照组患者(P<0.05);C1qAb阳性的SLE患者肾损发生率、活动性狼疮发生率及抗dsDNA抗体的阳性率均高于C1qAb阴性患者(P<0.05);C1qAb与SLEDAI活动性评分、抗核小体抗体(anti-nucleosome antibody,AnuA)及抗dsDNA抗体呈正相关(P<0.05)。结论抗C1q抗体对SLE的诊断和疾病活动性判断有重要价值;抗C1q抗体参与了SLE肾脏损害的发病机制。     

Autoantibodies against monomeric C-reactive protein in sera from patients with lupus nephritis are associated with disease activity and renal tubulointerstitial lesions

Serum levels of C-reactive protein (CRP) often remain low despite high disease activity in systemic lupus erythematosus (SLE). Sera from 96 patients with renal biopsy-proven active lupus nephritis, 24 of 96 patients in remission, and 49 patients with SLE with negative urinalysis (nonrenal SLE) was collected. Immunoglobulin G autoantibodies against monomeric CRP (mCRP) were screened by enzyme-linked immunosorbent assay with purified human CRP. Associations with clinical features, pathological data, and laboratory findings were investigated. The prevalence of mCRP autoantibodies in active lupus nephritis (57/96, 59.4%) was significantly higher than that in patients with SLE without clinical evidence of kidney involvement (20/49, 40.8%, p = 0.034). For the 13 patients with positive mCRP autoantibodies and sequential sera, their positive mCRP autoantibodies in active phase turned negative in remission (13/13, 100%). Patients with mCRP autoantibodies had significantly higher SLEDAI scores than patients without mCRP autoantibodies (18.3 +/- 5.2 vs 15.8 +/- 4.0, p = 0.013), who were more likely to experience acute renal failure (14/55 vs 2/33, p = 0.022), oral ulcer (15/57 vs 3/39, p = 0.022), and delayed activated partial thromboplastin time (18/52 vs 2/38, p = 0.001). Positive correlations between levels of mCRP autoantibodies and semiquantitative scores of renal histologic features were first observed in lupus nephritis as follows: interstitial inflammation (r = 0.328), tubular atrophy(r = 0.276), interstitial fibrosis (r = 0.211), and chronicity index score (r = 0.243). Autoantibodies against mCRP are prevalent in patients with lupus nephritis and are associated with disease activity and renal tubulointerstitial lesions.     

CD134 expression on CD4+ T cells is associated with nephritis and disease activity in patients with systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is characterized by a deviation of the immune system that involves T cell-dependent autoantibody production. The aim of this study was to investigate the role of co-stimulatory markers on T cells in this disease. Twenty-eight patients with SLE as defined by the American College of Rheumatology (ACR) criteria and 11 healthy controls were included into the study. Eleven patients had biopsy-proven lupus nephritis while 17 patients had no clinical evidence of lupus nephritis. Clinical disease activity was assessed according to the systemic lupus erythematosus disease index (SLEDAI). CD4+ T cell populations in the peripheral blood were analysed for the expression of co-stimulatory markers CD45RO, CD70, CD80, CD86, CD137, CD137L, CD134, CD152, CD154 and ICOS. SLE patients showed an increased frequency of peripheral CD4+ T cells expressing high levels of CD80, CD86 and CD134 compared to healthy controls (7.1 +/- 1.5% versus 1.7 +/- 0.9%; P < 0.005; 2.3 +/- 0.4% versus 1.0 +/- 0.2%; P = 0.008, 20.2 +/- 2.0% versus 10.6 +/- 1.9%; P < 0.005, respectively). Significantly higher levels of CD80 on CD4+ T cells were detected in SLE patients with lupus nephritis compared to patients without nephritis (11.9 +/- 3.3% versus 4.0 +/- 0.7%; P < 0.005). There was an increased presence of CD134+ CD4+ cells in SLE patients with lupus nephritis (27.5 +/- 4.0% versus 15.5 +/- 1.3%; P < 0.005). CD80 and CD134 expression was significantly correlated with SLEDAI (r = 0.42, P = 0.03; r = 0.56, P < 0.005). Co-stimulatory molecules on CD4+ T cells are associated with renal disease and disease activity in patients with systemic lupus erythematosus.     

Monocyte response to Th1 stimulation and effector function toward human mesangial cells are not impaired in patients with lupus nephritis

Monocytes/macrophages activated by Th1 stimulation such as interferon-gamma (IFN-gamma) and CD40 ligand (CD40L) infiltrate the kidney and play a critical role in the progression of lupus nephritis (LN). We examined the monocyte response to Th1 stimulation and their effector function toward activating renal resident cells in patients with LN. Following stimulation with IFN-gamma granulocyte macrophage-colony stimulating factor (GM-CSF)/recombinant CD40L the production of tumor necrosis factor-alpha and IL-12 p70 by PBMC was significantly higher in LN patients. In coculture experiments employing activated monocytes and human mesangial cells, there was a trend toward higher monocyte chemoattractant protein-1 production by lupus monocytes compared to normal controls. Basal expression of CD40, ICAM-1, and STAT-1 was significantly higher in monocytes from LN patients, suggesting ongoing activation. Monocyte response to IFN-gamma, as accessed by intercellular adhesion molecule-1 upregulation and phosphorylation of STAT-1, was comparable between the two groups. Thus, in contrast to earlier reports, Th1-dependent monocyte activation is not impaired. In this disease activated monocytes appear to be fully capable of inducing renal injury.     

LRCH1维持狼疮肾炎模型小鼠调节性T细胞的功能

目的 探讨富含亮氨酸的重复序列和钙调理蛋白同源域1(LRCH1)在狼疮肾炎模型小鼠调节性T细胞(Tregs)中的表达和功能.方法 6月龄C57BL/6小鼠和MRL/lpr小鼠,用流式细胞仪分选脾脏和肾脏内的CD45+CD4+CD25+CD127low Tregs,RT-qPCR检测Tregs LRCH1 mRNA水平....     

HMGB1+ microparticles present in urine are hallmarks of nephritis in patients with systemic lupus erythematosus

Non‐classical monocytes infiltrate the kidney parenchyma and participate in tissue damage in patients with lupus nephritis (LN). Circulating microparticles (MPs) seem to play critical roles in the activation of monocytes in systemic lupus erythematosus (SLE) patients. This study aims to characterize the phenotypes of MPs and monocyte subsets in LN patients and to determine their potential to discriminate between SLE patients with and without LN. Blood and urine samples from SLE patients were collected. In monocyte subsets from whole blood samples several phenotypic markers were evaluated. MPs were isolated from platelet‐poor plasma and urine by centrifugation. This phenotypic marker characterization was performed using multiparametric flow cytometry. We observed that patients with active LN have lower counts of non‐classical monocytes than do those without renal involvement. All monocyte subsets exhibited lower expression of CX3CR1 and ICAM‐1 in LN than in patients without LN. High frequencies of MP‐HMGB1⁺ and MP‐HLA‐DR⁺ were detected in circulation and urine of LN patients. Although MP‐HMGB1⁺, MP‐HLA‐DR⁺, and MP‐CX3CR1⁺ from urine were able to discriminate between patients with and without LN, only urinary MP‐HMGB1⁺ were different between patients with active and inactive LN. Therefore, these vesicles may be useful as biomarkers of LN.     

脑肿瘤患者胰岛素样生长因子-1水平的研究

目的探讨血清胰岛素样生长因子（IGF）-1水平与脑肿瘤的关系,垂体瘤、颅咽管瘤手术与血清IGF-1水平变化是否存在联系。方法检测193例脑肿瘤患者血清及61例术后相应患者血清IGF-1水平,用SPSS16．0统计软件进行分析。结果①恶性脑肿瘤组与良性脑肿瘤组间的差异有统计学意义（P〈0．05）,侵袭性垂体瘤组与非侵袭性垂体瘤组间差异有统计学意义（P〈0．05）;②颅咽管瘤手术前后血清IGF-1水平差异有统计学意义（P〈0．05）,侵袭性与非侵袭性垂体瘤手术前后血清IGF-1水平差异无统计学意义（P〉0．05）。结论①IGF-1在良性脑肿瘤和恶性脑肿瘤患者血清中表达的水平存在差异,可帮助临床对肿瘤的恶性程度作出早期的判断;②颅咽管瘤患者手术前后血清IGF-1水平存在差异,对患者的预后判断具有参考价值。     

CR1 stump peptide and terminal complement complexes are found in the glomeruli of lupus nephritis patients

The number of CR1 on podocytes is reduced in nephropathies with severe glomerular damage, especially in the diffuse proliferative glomerulonephritis (DPGN) of systemic lupus erythematosus (SLE). Reduction of CR1 number on erythrocytes is due to proteolysis of CR1 by macrophage proteases activated by the reaction of their complement receptors, which leaves a ‘CR1 stump peptide’ on the erythrocyte [1]. In the present study, we demonstrated the presence of the terminal complement complex (TCC) and the CR1 stump in histological sections of biopsies from patients with SLE by the indirect immunoperoxidase technique. Less severe glomerular lesions presented TCC deposits mainly in the mesangium (mesangial pattern). In lupus nephritis, with more severe glomerular damage, TCC deposits were detected both in the mesangium and in the capillary loops with podocyte involvement (mixed pattern). Patients with highly active DPGN presented a marked reduction of intact podocyte CR1 receptors in association with increased reactivity to the anti-CR1 stump antibody and with glomerular TCC deposits of mixed histological pattern. These results suggest that the decrease in the number of podocyte CR1 receptors in severe glomerular lesions of SLE may be due to a local proteolytic activity associated with activation and deposition of TCC.