Reduced Nitric Oxide Bioavailability In a Baboon Model of Shiga Toxin Mediated Hemolytic Uremic Syndrome (HUS)

Background. Although there is agreement that post-diarrheal hemolytic uremic syndrome (D + HUS) is caused by Shiga toxin (Stx)–producing E. coli, little is known about factors that mediate the host response to these toxins and potentially contribute to pathogenesis. Nitric oxide (NO) is a candidate mediator by virtue of its antiplatelet and renal vasodilatory properties. Methods. We used a baboon model of HUS to measure plasma and urinary NO metabolites and expression of NO synthase (eNOS and iNOS) in renal tissue following the intravenous administration of Stx-1. Results. Plasma concentrations through 60 hours of observation did not differ significantly from controls. Urinary values (indexed against urinary creatinine) tended, however, to rise during the initial 12 hours following administration of Stx-1. This was followed by a sustained reduction that coincided with the development of hemolytic anemia (schistocytosis) and other features of HUS. However, immunohistochemical staining for eNOS and iNOS in tissue obtained immediately after death at a median of 59 hours showed similar levels in control and Stx-treated animals, despite the presence of a florid thrombotic microangiopathy and tubular injury in the Stx-treated group. Conclusion. We propose that urinary NO metabolite reduction was due to NO inactivation subsequent to its avid binding to free hemoglobin released from lysed red blood cells, and that this contributed to the acute renal failure by facilitating vasoconstriction and platelet aggregation and adhesion within the renal microvasculature.     

Lupus nephritis associated with positive MPO-ANCA in a patient with underlying autoimmune hemolytic anemia

A 19-year-old female was admitted with general malaise and systemic edema. She had been diagnosed as having autoimmune hemolytic anemia (AIHA) eight years earlier and was successfully managed with oral prednisolone. During the current admission, she was diagnosed as having systemic lupus erythematosus (SLE) based on the presence of renal involvement, hematological abnormalities, and antinuclear and anti-double-stranded DNA antibodies, along with a recurrence of AIHA; her serology revealed a high myeloperoxydase-antineutrophil cytoplasmic antibody (MPO-ANCA) titer. She was treated with prednisolone (50 mg day⁻¹), but her renal function started to deteriorate. She responded to treatment with hemodialysis, plasmapheresis, and methylprednisolone pulse therapy; her MPO-ANCA titer and renal function improved. Treatment with intravenous cyclophosphamide gradually suppressed her AIHA and SLE activity. A renal biopsy revealed a diffuse proliferative lupus nephritis (class IV-G (A)) with necrotizing crescentic glomerulonephritis that was presumed to be associated with MPO-ANCA. The association of MPO-ANCA with SLE in this refractory case is discussed.     

结直肠癌患者手术前后血清中IL-6和NO水平的临床意义

目的探讨结直肠癌患者手术治疗前后血清中IL-6和NO水平变化及其临床意义。方法对40例结直肠癌患者分别采用酶联免疫分析法和化学比色法测定手术前后血清中的IL-6和NO水平,并与10名正常健康人作比较。结果结直肠癌患者手术前血清中IL-6水平高于正常人（P〈0.05）,NO水平明显低于正常人（P〈0.01）;经手术治疗2周后,患者血清IL-6水平较治疗前降低（P〈0.05）,而NO水平明显升高（P〈0.05）。结论结直肠癌患者血清中IL-6和NO水平的变化,对病情和预后判断具有重要的临床意义。     

Glucose intolerance and hepatocellular carcinoma: recent findings for old diseases

In the last years, an increasing number of evidences on the influence of metabolic syndrome on the occurrence of hepatocellular carcinoma (HCC) have been developed. Type 2 mellitus diabetes (T2MD) has been found to increase the occurrence of primary liver tumors and to define a more aggressive carcinogenetic process. Furthermore, several preclinical and observational studies and a recent meta-analysis have shown that anti-diabetic drugs can modify the risk of HCC development in patients with T2DM. However, despite these evidences, underlying molecular mechanisms linking both pathological conditions have to be completely cleared yet. The study published by Gao et al. has found a possible molecular link between the two conditions, describing the predisposition to T2DM and HCC given by the haploinsufficiency of nuclear receptor coactivator 5 (NCOA5) in murine models. The authors have generated Ncoa5+/– (haploinsufficient) male mice and shown that 94% of male mutant mice developed HCC within 18 months of age, this in contrast with Ncoa5+/+ and Ncoa5+/– female mice. These results suggest that NCOA5 haploinsufficiency is linked to HCC development in male mice. Moreover, mutant male mice showed significantly elevated levels of fasting blood glucose and markedly decreased glucose tolerance and insulin sensitivity compared to Ncoa5+/+ littermates. This well-constructed work sheds light on the molecular link between T2DM and HCC and opens the way to further biological and clinical studies in the field of liver tumor prevention and treatment.     

Uneventful splenectomy and cholecystectomy in a patient treated with anti-interleukin-6 receptor antibody therapy

Introduction Interleukin-6 (IL-6) is a multifunctional cytokine that regulates various aspects of the immune responses, acute phase reactions, and hematopoiesis. In rodent models, IL-6 has been suggested to be one of the essential mediators for optimal acute phase responses to infection and tissue damage. However, in humans, the roles of IL-6 in acute phase responses after surgery remain poorly understood. Case report We present the first case report of successful splenectomy and cholecystectomy in a severe autoimmune-associated hemolytic anemia patient during treatment with a humanized anti-IL-6 receptor antibody. Discussion This unique case suggests that IL-6 is not an essential cytokine to safely perform surgical intervention and to prevent postoperative complications and that surgical intervention may not be contraindicated but can be selected as a therapeutic modality in patients treated with anti-IL-6 receptor antibody therapy.     

Fibrin Plugging as a Cause of Microcirculatory Occlusion During Photodynamic Therapy

An experimental system that allows the white light observation of rapid changes in vessels without disturbance by red laser light was used. Mice were injected with mono-l-aspartyl chlorin-e-6 (Npe-6) i.v. via the tail vein and were immediately exposed to laser light. White emboli were observed forming on the inside of the vessel walls within seconds after commencement of light exposure. Emboli adhered to vessel walls and caused vascular obstruction. Light microscopy of the exposed material using fibrin staining was performed. Electron microscopy on the same material was also carried out. The embolisation time was influenced by both drug dose and laser power. With low laser power, it took a long time to stop the blood flow. Fibrin staining revealed the white emboli to be composed of fibrin. Electron microscopy findings revealed damage to endothelial cells and platelet aggregation. This study suggests that two main mechanisms (direct cellular damage and vascular shut-down ) might actually be complementary and synergistic in the production of vascular lesions using photodynamic therapy. Paper received 28 February 1998; accepted after revision 8 October 1998.     

Urinary neutrophil gelatinase-associated lipocalcin in D+HUS: a novel marker of renal injury

Background: Diarrhea-associated hemolytic uremic syndrome (D+HUS) causes acute renal failure. Neutrophil gelatinase-associated lipocalcin (NGAL) is an early indicator of kidney injury. Objective: To determine if urinary NGAL excretion is a biomarker of severe renal injury and predicts the need for dialysis in D+HUS. Methods: Patients were randomly selected from among participants in the SYNSORB Pk trial. Urine samples were collected daily if available during the first week of hospitalization. NGAL levels were determined by ELISA. Results: 34 children, age 5.9±3.9 yr, were studied; ten (29%) required dialysis. Patients were categorized based on urinary NGAL concentration within five days of hospitalization - <200 ng/ml and ≥200 ng/ml. Twenty patients (58%) had increased urinary NGAL excretion. The severity of D+HUS at enrollment was similar in the two groups. However, children with increased urinary NGAL levels had higher peak BUN and creatinine concentrations (P<0.01) and required dialysis more often, 9/20 versus 1/14 (P=0.024) compared to children with normal excretion. Conclusion: The majority of patients with D+HUS have renal tubular epithelial injury, as evidenced by elevated urinary NGAL excretion. Urinary NGAL levels below 200 ng/ml within five days of hospitalization may be an adjunctive marker that defines less severe renal involvement.     

C REACTIVE PROTEIN IN PATIENTS WITH CHRONIC RENAL DISEASES

Base-line serum levels of plasma C-reactive protein (CRP) are predictive of future myocardial infarction and sudden cardiac death in apparently healthy subjects, suggesting the hypothesis that chronic inflammation might be important in the pathogenesis of atherothrombosis. CRP production is mediated by several inflammatory mediators: interleukin 6 (IL-6) is currently felt to be the major cytokine influencing the acute phase response. CRP and other acute phase proteins are elevated in dialysis patients and cardiovascular diseases represent the single largest cause of mortality in chronic renal failure patients. Little information is available, however regarding CRP and IL-6 plasma levels in pre-dialysis renal failure. Plasma CRP was determined by a modification of the laser nephelometry technique; IL-6 by immunoassay (RD System); and fibrinogen, serum albumin, cholesterol, triglycerides, hematocrit, white blood cell count, erythrocytic sedimentation rate (ESR) and urinary protein levels by standard laboratory techniques. Results were obtained in 102 chronic pre-dialysis patients whose mean age was 53 ± 5.8 years with a mean creatinine clearance (C_Cr) of 52 ± 37 mL/min). CRP was greater than 5 mg/L in 25% of the global population. CRP and IL-6 were 4.0 ± 4.6 mg/L and 5.8 ± 5.6 pg/mL, respectively and were not significantly correlated (r = 0.11, p = n.s.). CRP and IL-6 were however related with renal function (CRP versus C_Cr r = ?0.40 p < 0.001; IL-6 versus C_Cr r = ?0.45; p < 0.001). When patients were divided in two groups according to renal function, CRP resulted 7.4 ± 6.3 mg/L in the group of patients with a C_Cr lower than 20 mL/min (n = 32) and 2.76 ± 4.35 in the group of patients with a C_Cr higher than 20mL/min (n = 70) (p < 0.0001). CRP and IL-6 were positively related with ESR (r = 0.32 and 0.46 respectively). Serum albumin levels were not significantly different in the two groups of patients (3.2 ± 0.4 versus 3.0 ± 0.5 g/dL). CRP and serum albumin were not significantly related (r = 0.17). CRP and IL-6 correlated positively with ESR (r = 0.32 and 0.46 respectively). In pre-dialysis patients we have demonstrated an increase in both CRP and IL-6 that occurs as renal function decreases. These data provided evidence of the activation – even in the predialysis phase of renal failure – of mechanisms known to contribute to the enhanced cardiovascular morbidity and mortality of the uremic syndrome.     

急性胰腺炎合并肝损伤时IL-6、氧自由基变化及IL-2、川芎嗪干预的实验研究

目的：探讨IL－6、氧自由基在急性胰腺炎（acute pancreatitis，AP）合并肝损伤中的作用，及重组人白介素－2（IL－2）、川芎嗪的治疗价值。方法：SD大鼠112只，随机分为14只，每组8只，5％牛磺胆酸钠逆行胰胆管内注射诱发大鼠AP动物模型，检测血浆IL－6、SOD、MDA、ALT、AST、LDH、LIP、AMY，并观察肝、胰病理变化。结果；①AP组血浆AMY、LIP、ALT、AST、LDH明显升高（P＜0．05或0．01），镜下可见胰腺水肿、炎细胞浸润、坏死1肝脏肝窦充血、细胞浊肝及坏死，且损伤程度随时限延长而加重；②AP各组血浆IL－6明显升同（P＜0．01）；③AP各组MDA明显高（P＜0．01）、SOD明显降低（P＜0．01）。④IL－2治疗组、川芎嗪治疗组有IL－2、川芎嗪联合且与NS组比较血浆IL－6、MDA水平明显下降（P＜0．05），SOD明显升高（P＜0．05），胰、肝病理损害程度减轻，并且AMY、LIP、ALT、AST、LDH均明显降低（P＜0．05），平均存活时间明显延长（P＜0．05）；联合应用组降低IL－6、MDSA水平和减轻胰腺坏死优于单药组。结论：①IL－6、氧自由基在急性胰腺炎合并肝损伤程度和明显升同，起损伤作用，而SOD明显降低，其保护作用减弱。检测血浆IL－6、MDA、SOD可作为判断AP合并肝脏损伤程度和预后的指标；②大鼠急性胰腺合并肝损伤过程中应用IL－2、川芎嗪显示出良好的效果，联合应用于亿于这两药的单独应用。     

Effect of interleukin 6 on basal and FSH-stimulated secretion of estradiol and progesterone by human granulosa cells in vitro

Aim: To determine the effects of interleukin-6 (IL-6) on the secretion ofestradiol and progesterone by human granulosa cells in vitro. Methods:Granulosa cells were obtained from infertile patients undergoing IVF ETtreatment and cultured with serum-free supplemented HAM's F10 medium.In the absence or presence of FSH, granulosa cells were treated with differ-ent concentrations of gene recombinant human iterlukin-6 (rhIL-6). Themedia were collected after 24, 48, 72 and 96 h and assayed for estradiol andprogesterone. IL-6 and R mRNA was determined by means of RNA slotblot. Results: IL-6 had a significant inhibitory effect on estradiol secre-tion, especially in the presence of FSH. IL-6 inhibited the FSH-stimulatedprogesterone secretion, but not the basal progesterone release. The inhibi-tion shows certain degrees of dose- and time-dependency. Conclusion:IL-6 participates in the regulation of ovarian function through its inhibitoryeffect on FSH-stimulated steroidogenesis by granulosa cells.     

m6A RNA methylation regulators play an important role in the prognosis of patients with testicular germ cell tumor

BackgroundN6-methyladenosine (m6A) is found to be associated with promoting tumorigenesis in different types of cancers, however, the function of m6A-related genes in testicular germ cell tumors (TGCT) development remains to be illuminated. This study aimed to investigated the prognostic value of m6A RNA methylation regulators in TGCT.MethodsWe collected TGCT patients’ information about clinicopathologic parameters and twenty-two m6A regulatory genes expression from The Cancer Genome Atlas (TCGA) database and Genotype-Tissue Expression (GTEx). We analyzed the differentially expressed m6A RNA methylation regulators between tumor tissues and normal tissues, as well as the correlation of m6A RNA methylation regulators. By using Cox univariate analysis, last absolute shrinkage and selection operator (LASSO) Cox regression algorithm and Cox multivariate proportional hazards regression analysis, a risk score was constructed based on a TCGA training cohort, and further verified in the TCGA testing cohort. Then, univariate and multivariate Cox regression analyses were used to evaluate the relationship between risk score and progression-free survival (PFS) in TGCT. Finally, the six-gene risk score was further verified by two gene expression profiles ({"type":"entrez-geo","attrs":{"text":"GSE3218","term_id":"3218"}}GSE3218 and {"type":"entrez-geo","attrs":{"text":"GSE10783","term_id":"10783"}}GSE10783) as an independent external validation cohort.ResultsDistinct expression patterns of m6A regulatory genes were identified between TGCT tissues and normal tissues in TCGA and GTEx datasets. To predict prognosis of TGCT patients, a risk score was calculated based on six selected m6A RNA methylation regulators (YTHDF1, RBM15, IGF2BP1, ZC3H13, METTL3, and FMR1). Additionally, we found significant differences between the high-risk and low-risk groups in serum marker study levels and histologic subtype. Univariate and multivariate analysis indicated that high risk score was associated with unfavorable PFS. Ultimately, the risk score was further verified by two gene expression profiles ({"type":"entrez-geo","attrs":{"text":"GSE3218","term_id":"3218"}}GSE3218 and {"type":"entrez-geo","attrs":{"text":"GSE10783","term_id":"10783"}}GSE10783).ConclusionsBased on six selected m6A RNA methylation regulators, we developed a m6A methylation related risk score that can independently predict the prognosis of TGCT patients, and verify the prediction efficiency in TCGA and GEO datasets. Patients in high-risk group were associated with serum tumor marker study levels beyond the normal limits, non-seminoma, and unfavorable survival time. However, further prospective experiments should be carried out to verify our results.     

大鼠烧伤后库普弗细胞在促炎细胞因子产生中的作用

目的　观察大鼠严重烧伤后早期 ,库普弗细胞在肿瘤坏死因子α(TNFα)、白细胞介素(IL) 1β、IL 6产生中的作用。方法　观察 (1)烧伤血清对体外培养的大鼠库普弗细胞分泌TNFα、IL 1β、IL 6的刺激作用 ;(2 )烧伤后大鼠库普弗细胞的细胞因子mRNA表达变化 ;(3)应用库普弗细胞特异性抑制剂三氯化钆后 ,烧伤大鼠血浆内细胞因子含量变化。　结果　烧伤血清能刺激库普弗细胞释放TNFα、IL 1β、IL 6 ;大鼠烧伤后库普弗细胞TNFα、IL 1β、IL 6mRNA表达量显著升高 ;预先抑制库普弗细胞的活性 ,烧伤后血浆TNFα、IL 1β、IL 6水平均显著降低 ,分别为烧伤组的 34.71%、36 99%、33.70 %。结论　库普弗细胞是大鼠烧伤后血浆中TNFα、IL 1β、IL 6的主要来源     

系统性红斑狼疮伴自身免疫性溶血性贫血的临床分析

目的:了解自身免疫性溶血性贫血的系统性红斑狼疮(SLE)患者与无溶血性贫血的系统性红斑狼疮患者临床、实验室检查方面的异同。方法:将40例SLE患者分为2组,一组为抗人球蛋白试验(Coombs)阳性,另一组为Coombs阴性,进行临床症状及实验室检查比较。结果:Coombs阳性组中免疫球蛋白(IgG)、补体C_3、球蛋白(GLB)、总蛋白(TP)较非阳性组明显降低(P<0.05)。结论:Coombs阳性组患者因免疫机能紊乱产生自身抗体引起溶血,同时其病情活动程度高。