ABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of IL-17 in the overproduction of autoantibodies and IL-6 overexpression by peripheral blood mononuclear cells (PBMC) of lupus nephritis (LN) patients.</p><p><b>METHODS</b>Fifteen consecutively hospitalized LN patients were selected as subjects and 15 healthy adults as normal controls. PBMC were obtained by Ficoll density gradient centrifugation. IgG, anti-dsDNA antibody and IL-6 protein levels were assessed using enzyme-linked immunosorbent assays (ELISA) on the supernatant of cultured PBMC of LN patients or normal controls. IL-6 mRNA levels in PBMC were measured using reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>In medium culture, IgG, anti-dsDNA and IL-6 protein levels of the supernatant of PBMC from LN patients were significantly higher than those from normal controls (1492.1 +/- 73.2 ng/ml vs 636.7 +/- 51.9 ng/ml for IgG, 306.6 +/- 53.7 IU/ml vs 95.8 +/- 11.6 IU/ml for anti-dsDNA and 50.92 +/- 15.92 ng/ml vs 1.77 +/- 0.73 ng/ml for IL-6, all P < 0.001). In LN patients, IgG, anti-dsDNA and IL-6 protein levels were higher in the supernatants of PBMC in the IL-17-stimulated culture than the medium culture, but in normal controls, only the IL-6 protein levels were significantly higher. The increase in IgG, anti-dsDNA and IL-6 protein levels induced by IL-17 was dose-dependent and could be completely blocked by IL-17 monoclonal antibody mIgG(28) and partially blocked by dexamethasone. Similarly, IL-6 mRNA overexpression of PBMC in LN patients or normal controls induced by IL-17 was both dose- and time-dependent. During medium culture, IL-6 mRNA levels in LN patients were significantly higher than those in normal controls (1.80 +/- 0.11 vs 0.36 +/- 0.07). During stimulation with IL-17, IL-6 mRNA levels in LN patients were higher than those in normal controls (3.21 +/- 0.24 vs 1.30 +/- 0.14, P < 0.05) and also significantly higher when comparing the stimulated culture with the medium culture either in LN patients or normal control.</p><p><b>CONCLUSIONS</b>IL-17 may play an important role in the pathogenesis of LN through the induction of IgG, anti-dsDNA overproduction and IL-6 overexpression of PBMC in LN patients.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Antibodies, Antinuclear , Autoantibodies , Immunoglobulin G , Interleukin-17 , Pharmacology , Interleukin-6 , Genetics , Leukocytes, Mononuclear , Metabolism , Lupus Nephritis , Allergy and Immunology , RNA, MessengerABSTRACT
<p><b>OBJECTIVE</b>To determine the in vitro expression of interleukin-12 (IL-12) and its effect on signal transducers and activators of transcription (STAT) signaling molecules in peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE).</p><p><b>METHODS</b>Peripheral blood mononuclear cells in 39 patients with definite systemic lupus erythematosus and 11 healthy volunteers were collected. Expression of IL-12 P40mRNA in PBMCs was determined with reverse transcription-polymerase chain reaction (RT-PCR). Quantity of IL-12 protein supernatant was measured by enzyme-linked immunosorbent assay (ELISA). The levels of phosphorylated STAT3 and STAT4 signaling molecules in PBMCs were detected by immunoblot.</p><p><b>RESULTS</b>Levels of IL-12 protein and mRNA expression in patients with active or inactive SLE were significantly higher than those in controls. Phytohemagglutinin (PHA ) may promote the expression of IL-12. IL-12 alone induced the phosphorylation of STAT3 and STAT4 in PBMCs from patients with SLE, especially in active SLE. However it had no obvious effect on normal PBMCs. Phosphorylated STAT3 and STAT4 might be observed in normal PBMCs treated with IL-12 plus PHA.</p><p><b>CONCLUSION</b>IL-12 is produced aberrantly in patients with SLE. IL-12 might exert its biological role in SLE via the aberrantly phosphorylated STAT3 and STAT4 signaling molecules.</p>
Subject(s)
Adolescent , Adult , Humans , Middle Aged , Cells, Cultured , DNA-Binding Proteins , Metabolism , Interleukin-12 , Blood , Genetics , Leukocytes, Mononuclear , Metabolism , Lupus Erythematosus, Systemic , Metabolism , Phosphorylation , RNA, Messenger , STAT3 Transcription Factor , STAT4 Transcription Factor , Trans-Activators , MetabolismABSTRACT
0 05).Of patients with lupus renal lesions 86% had a positive LBT (+LBT),which was significantly higher than that of patients without renal lesions(37%) ( P
ABSTRACT
Objective To study clinical feature,pathology,treatment and outcome of acute renal failure(ARF) in idiopathic nephrotic syndrome(IDNS).Methods History-taking of 38 cases were carried out all detailly 24-hour urine protein,biochemistry markers,hemorheology,renal histopathology were detected.Results The study fandings that 82 percent of patients were less than 40 years old,76 3 percent were male,74 percent(28 patients) were non-oliguria and 52 6 percent were minimal change glomerulonephritis through the renipuncture,most of the cases had edema of interstitium, by give prednisone of standard dosage,symptomatic treatment or dialysis the satisfying efficacy was got.Conclusions The main features of IDNS complicated with ARF are usual,it is non-oliguria type in clinic minimal glomerulonephritis in pathology,and deficient blood volume leads to occurrence of ARF,but the renal function could be recovered by give prednisone of routine dosage,symptomatic treatment and hemodialysis therapy.
ABSTRACT
AIM:To determine whether human peritoneal mesothelial cells express CD40. METHODS:Human peritoneal mesothelial cells (HPMC) were harvested from spent peritoneal dialysis effluent and maintained under defined in vitro conditions. Expression of CD40, CD40L and intercellular adhesion molecule-1 (ICAM-1) on HPMC under normal culture or stimulation with interferon-γ (IFN-γ), tumor necrosis factor-α(TNF-α), interleukin(IL)-1 and LPS were detected by FACS analysis. The relationship between CD40 expression and ICAM-1 expression on HPMC was analyzed.RESULTS:HPMC cultured in vitro expressed CD40 constitutively. The surface expression of CD40 was markedly up-regulated following stimulation with IFN-γ, but not with TNF-α, IL-1 and LPS. CD40L expression on HPMC was not detected. The expression of ICAM-1 on HPMC was significantly increased by stimulation with IFN-γ, TNF-α, IL-1, LPS, respectively; the most effective inducer on ICAM-1 expression was IFN-γ. The CD40 expression co-localizes with the expression of ICAM-1 and there was positive correlation between CD40 and ICAM-1 expression on HPMC. CONCLUSION:HPMC functionally expressed CD40.
ABSTRACT
AIM:To investigate the regulatory role of nuclear factor κB (NF-κB) in the expression of interleukin-6 in mesangial cells (MC) induced by interleukin-1β.METHODS:Activation of NF-κB was measured by electrophoresis mobility shift assay (EMSA). RT/PCR and ELISA were used to detect IL-6 mRNA expression and IL-6 production, respectively.RESULTS:rhIL-1β could rapidly stimulate the activation of NF-κB in MC, and increase the expression of IL-6 mRNA and protein. PDTC, one of the inhibitor of NF-κB, could inhibit the expression of IL-6 in mRNA and protein in MC stimulated by rhIL-1β.CONCLUSION:IL-6 expression induced by IL-1β may be regulated by NF-κB in MC, NF-κB may modulate the immune-inflammatory reaction in glomerular disease.
ABSTRACT
【Objective】To observe the expression of PI3-K phosphorylated products and elucidate the correlation between PI3-K phosphorylated products and Th2 cytokine in peripheral blood mononuclear cell (PBMC).【Methods】14 patients with active lupus nephritis and 12 controls were selected,PI3-K phosphorylated products were detected by immunoprecipitation and Western blotting,RT-PCR was used to observe interleukin-6 mRNA and interleukin-10 mRNA expression.【Results】In either spontaneous condition or stimulated by anti-CD3 antibody,the expression of PI3-K phosphorylated products in patients with active lupus nephritis were higher than those of the controls(1.14±0.23 vs 0.46±(0.12,P=0.023;2.09±0.63 vs 0.65±0.14,P=0.016).The expression of PI3-K phosphorylated products in active lupus nephritis showed a positive correlation with interleukin-6 mRNA and interleukin-10 mRNA (r=0.652,P=0.008;r=0.718,P=0.007).PY294002,one of specific inhibitor of PI3-K,inhibited significantly the expression of interleukin-6 mRNA(2.32±0.51 vs 0.57±0.15,P=0.009) and interleukin-10 mRNA (1.71±0.33 vs 0.67±0.11,P=0.006) in stimulated PBMC in active lupus nephritis.【Conclusion】PI3-K can involve in the pathogenesis of lupus nephritis by inducing the overexpression of interleukin-6 and interleukin-10.
ABSTRACT
ObjectiveTo investigate the clinical features of acute myocardial infarction in CAPD patients.MethodsCompare the clinical and laboratory data of 39 patients with CAPD and AMI for the first time (PD group),including clinical symptoms,signs,electrocardiogram changes,myocardial enzyme changes and in hospital mortality,with those of 50 patients without renal insufficient but with AMI for the first time (C group).Diagnosis of AMI was made in hospital.ResultsCompared to C group, the symptoms presented at first in PD group were more atypical and the patients went to see doctor more lately after AMI.Atypical electrocardiogram changes were more in PD group than in C group (61.5% vs 26.0%,P
ABSTRACT
Objective To investigate the value of dialysis and ethics of withdrawal dialysis.Method Through analysis of some typical cases of ESRD patients,to investigate the value of dialysis.Conclusion A lot of ESRD patients can survive through dialysis,thanks to the improvement of medical science in last 30 years.On the other hand,it can also keep some patients with no living calculation surviving,becoming the burdensome of families,hospitals and societies.Our object is to discuss the ethics of withdrawal dialysis in our country.
ABSTRACT
Objective:To detect the activation of NF AT and AP 1 in peripheral blood mononuclear cells(PBMCs) in patients with systemic lupus erythematosus and investigate their relationship with serum ANA,anti dsDNA antibody,ESR,CRP and C 3.Methods:Activation of NF AT and AP 1 were detected by electrophoretic mobility shift assay(EMSA).Results:①Activation of NF AT in PBMC in active and inactive SLE patients was increased when compared with normal controls,respectively,and the activation of NF AT in active patients was higher than that of inactive ones.②Elevated activation of AP 1 in PBMC in active SLE patients was found as compared with normal controls,and there was no significant difference in activation of AP 1 between inactive patients and normal control.③Activation of NF AT in SLE patients with positive anti dsDNA antibody was higher than that of patients with negative anti dsDNA antibody,and there was not different positively in AP 1 activation between above two patient groups.④Activation of NF AT in PBMC was related positively with CRP and SLEDAI,respectively,but no correlation with ESR,ANA,C 3.There was no relation between the activation of AP 1 and above clinical parameters.Conclusion:The abnormal expression of NF AT and AP 1 indicates disturbed intracellular signaling in SLE,which is suggested that alternations in activation of transcription factors may contribute to the pathogenesis of SLE.Activation of NF AT may be used as a predicator of SLE disease activity.
ABSTRACT
AIM: To investigate the influence of dialysate on the cell morphology and the peritoneal membrane function in chronic peritoneal dialysis rats. METHODS: 40 SD rats were divided randomly into 4 groups. Except control group, other groups daily received infusion of 20 mL dialysate (4.25% dialysate(HG), 1.50% dialysate(LG), Riger's solution(RG)) respectively for 8 weeks. The peritoneal membrane function was investigated by peritoneal transport test, and the rat peritoneal mesothelial cells(PMCs)morphology was analyzed by the cell imprints. RESULTS: The intraperitoneal volume and net ultrafiltration in HG and LG groups were significantly lower, with D/P_(urea) significantly higher, and C_(urea) after 4 h of dialysis significantly lower than that in RG and control groups. The density of cell population from analysis of cell imprints in HG and LG groups was significantly lower, but the mean surface area were significantly larger than that in RG and control groups. These change had no difference between HG and LG group. Using the high glucose dialysate for 8 weeks significantly decreased the ultrafiltration volume ,which was significantly relate to the increasing of cell surface area (r=-0.896,P
ABSTRACT
AIM: To investigate the role of CD134 (OX40) and NF-?B in the pathogenesis of lupus nephritis (LN). METHODS: Renal in situ CD134 and NF-?B expression were examined in 40 biopsy specimens from LN patients by immunohistochemistry and microwave-based immunohistochemistry, respectively. The relationship between expression of CD134 and NF-?B was analyzed. RESULTS: The expression of glomerular and tubular CD134 and NF-?B in LN were higher than that in normal control, especially in class Ⅳ LN, where there was intense staining of endothelial cell, distal tubules, and interstitial mononuclear cell. The CD134 expression of glomerular and tubular was closely related to NF-?B expression, respectively ( r=0.5542,P
ABSTRACT
AIM: We have shown that intraperitoneal (IP) addition of hyaluronan could increase peritoneal fluid removal by decreasing peritoneal fluid absorption rate. In this study, we investigated the impact of chronic use of hyaluronan on peritoneal membrane transport characteristics. METHODS: Twenty male SD rats received daily IP injection of 25 mL 4.25% glucose dialysis solution without (HP, n=8) or with 0.025% hyaluronan (HA, n=6) for one week, another six rats did not receive any peritoneal injection. Twenty-four hours after the last injection, a 4h dwell study using 25 mL 4.25% glucose dialysis solution with IP volume marker and frequent dialysate and blood samplings were performed in each rat. RESULTS:IPV was significantly higher in the HA group as compared to the HP group (ANOVA repeated measurement, P
ABSTRACT
AIM: To investigate the association of gene polymorphism at position 196 of tumor necrosis factor receptor Ⅱ (TNFRⅡ) with systemic lupus erythematosus (SLE) in Chinese, and establish recombinant retroviral vector to analyze the function of the TNFRⅡ 196M/R. METHODS: The genotype at position 196 of TNFRⅡ was determined by PCR-RFLP in 106 SLE patients and 119 healthy controls in china. Human TNFRⅡ196M cDNA were amplified by PCR and cloned into PMD18-T vector. Then, PMD18-TNFRⅡ196R was induced by site-directed mutagenesis. The recombinant T vector, PMD18-TNFRⅡ196M and PMD18-TNFRⅡ196R, were subcloned into retroviral vector PLXSN. Both normal and variant were transfected into rat mesangial cell. The effects of TNF? on production of sTNFRⅡ and IL-6 were study by ELISA. RESULTS: (1) The frequency of TNFRⅡ196R allele was significantly higher than those in controls (35.2% vs 14.3%, P
ABSTRACT
AIM:To study the effects and mechanism of recombinant human defensin ?1 on cell proliferation in cultured rat glomerular mesangial cells.METHODS:The influences of defensin ?1 at various concentrations on rat 1097 mesangial cell line cultured in vitro were evaluated with MTT assay.The different concentrations of U0126,signal-regulated protein kinase(MEK)inhibitor,were added into the culture mediums of mesangial cells to do blocking test.Incubated with a final concentration of 3 mg/L defensin ?1,the phosphorylation of extracellular signal regulated kinase(ERK)1/2 and type IV collagen of mesangial cells in different times were evaluated by Western blotting.RESULTS:Defensin ?1 at 3-20 mg/L enhanced proliferation of rat glomerular mesangial cells.The incubation times for the maximum effect on proliferation was 12 h(P20 mg/L decreased cell proliferation.The cell proliferation induced by defensin ?1 was inhibited by U0126.Stimulation of the cells with defensin ?1 at concentration of 3 mg/L for 5 minutes induced a maximum effect on a ratio of phosphorylation of ERK1/2 to total ERK.After 12 h incubation with defensin ?1,an increase in type IV collagen was observed by Western blotting and continued to increase at 24 h and 48 h(P
ABSTRACT
To investigate the best intennittence and reasonable dosage of CDC pulse therapy on different activity degree of lupus nephritis (LN). Methods 96 severe LN cases were divided into three groups. Croup A; CTX pusle treatment (IV-CTX), once 2 weeks, 8- 12 mg'kg'Vd for two days; Group B: IV-CTX once a month, 0.5 - 1.0/m2; Group C: IV-CTX once 3 months, 0.5 - 1.0/m2. Prednisone was given to three groups simultaneously. Results The time taking effect was shorter significantly in Group A than that in Group B and C. Remission rate was higher significantly in Group A than that in Group B and C ( P 0. 05). Intermittence of 15 cases (23.8%) from Group Ek C was changed to once 2 weeks and then the disease activity had been controlled since these patients became worse during the treatment. Intennittence of 3 patients (9% ) from Group A was delayed a week because of the decreasing of WBC counting. There was no significantly difference between three groups in side effect and its incidence. Conclusion A reasonable IV-CTX should be choosen according to the disease activity. IV-CTX 8 - 12 mg'kg'Vd for two days, once 2 weeks should be used for acute and severe LN. When disease becomes mild, IV-CTX can be changed to 0.5 - 1.0/m2, once a month. After LN activity is controlled basically, 0.5- 1.0/m2, once a month is recommended.
ABSTRACT
Objective In order to investigate the reversibility of uremia in patients with lupus nephritis(LN). Methods 37 uremic patients with IN who underwent dialysis were treated with a progressive protocol of combined glucocorticoid and cyclophosphamide (CTX) during the last 15 years. Results 83.8% of patients withdrawed from dialysis, with an average withdrawal period of 40.6 ?21.5 months, and with serum creatinine of 179.36 ?88.24 fjiaoVL at least half year after the withdrawal. Conclusion Uremic patients with LN who should be dialysed are not necessarily in end stage of their renal diseases, and those who are indicated for immunosuppressants and probable to be reversed should be treated properly and promptly.
ABSTRACT
Objective To determine whether peritoneal mesothelial cells express CD40 and to investigate potential mechanism by which CD40-CD40 ligand(CD40L) interaction may be involved in the inflammation of peritoneal membrane. Methods Rat peritoneal mesothelial cells (MC) were harvested from peritoneal cavity and maintained under defined in vitro conditions. Expression of CD40 on MC under normal culture or stimulation with interferon-?(IFN-?), tumor necrosis factor-?(TNF-?), interleukin(IL)-1 was examined by RT-PCR and FACS analysis. After activation with CD40 mAb, the expression of intercellular adhesion molecule-1 (ICAM-1) on MC was analyzed by FCAS. Results MC cultured in vitro expressed CD40 constitutively. The expression of CD40mRNA and CD40 protein up-regulated markedly following the stimulations with IFN-?, but not with IL-1, TNF-?. The expression of ICAM-1 on MC was significantly increased after activation of CD40 with IFN-? and CD40mAb. Conclusions MC functionally expresses CD40. The interaction of CD40 on MC and CD40L+ cells in peritoneal cavity may play an important role in peritoneal local defense and may be involved in the inflammation process of peritoneum.
ABSTRACT
Objective To define which hyaluronan synthase (HAS), of three hyaluronan synthesizing enzymes HAS-1, HAS-2, and HAS-3, is primarily responsible for hyaluronan synthesis and extracellular matrix/extracellular coat formation in human peritoneal mesothelial cells (HPMCs) . Methods As a prerequisite study, the expression of each HAS mRNA in cultured HPMCs was measured by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). Only the expression of HAS-2 and HAS-3 mRNA could be detected. The level of HAS-2 mRNA expression was about 10 fold higher than that of HAS-3. HAS-2 specific antisense oligonucleotide was then transfected into cultured HPMCs by lipofectamine. After 0, 8, 24, and 48 hours, the expression of HAS-2 mRNA was detected by RT-PCR and the extracellular coat was measured by particle exclusion test. Results 8 hours and 24 hours after transfection, the expression of HAS-2 mRNA in HPMCs decreased by 58% and 89% respectively; 48 hours after transfection, the expression of HAS-2 mRNA in HPMCs partially restored to 25% of the normal level. Correspondingly, 24 hours after transfection, the extracellular matrix/extracellular coat in HPMCs almost completely disappeared. However, as control, sense and reverse oligonucleotides showed no effect. Conclusion HAS-2 plays a leading role in HPMCs hyaluronan synthesis and the formation of extracellualr matrix/extracellular coat.