Elevated soluble receptor activator of nuclear factor-κB ligand and reduced bone mineral density in patients with adolescent idiopathic scoliosis

Generalized low bone mass and osteopenia in both axial and peripheral skeleton in adolescent idiopathic scoliosis (AIS) have been reported in literature. However, the exact mechanisms and causes of the bone loss in AIS are not identified yet. Therefore, this study examined the relationship between serum concentration of soluble receptor activator of nuclear factor-κB ligand (RANKL), serum level of osteoprotegerin (OPG) and bone mass in 72 patients with AIS and compared to those of 64 age- and gender-matched healthy controls. The mean lumbar spinal bone mineral density (LSBMD) and femoral neck BMD (FNBMD) in patients with AIS were decreased compared with that in control individuals, respectively (P = 0.0029 and P = 0.0192, respectively). The mean RANKL and RANKL to OPG ratio in patients with AIS were increased compared with that in control subjects, respectively (P = 0.0004 and P = 0.0032, respectively). The RANKL and RANKL to OPG ratios were negatively correlated to the LSBMD and serum OPG levels in both groups. Serum OPG levels were positively correlated to the LSBMD and FNBMD in both groups. These findings mean that the imbalance and the disturbed interaction of RANKL and OPG may be an important cause and pathogenesis in reduced BMD in AIS.     

Intra-alveolar IL-6 levels following burn and inhalation injury

Burn combined with inhalation injury is a major challenge and requires further study. Using a small-animal model, excretion of IL-6 was investigated during the first 6 h after exposure of rats to wood/polyvinyl chloride smoke, with and without concomitant skin burn. In controls, respirator therapy alone was found to release IL-6 into the serum and the alveolar space. These levels of IL-6 were reduced when associated with either inhalation injury or burn, but were increased when the traumas were combined. Thus, during the first 6 h of mechanical respiration the presence of burn or of inhalation injury seems to decrease IL-6 excretion, but a combination of these traumas reverses this effect.     

Osteoprotegerin and receptor activator of nuclear factor-kappaB ligand system in the early post-operative period of liver transplantation

Abstract: Background: The precise mechanism that leads to accelerated bone resorption in the early post‐transplant period remains unclear. Recent data suggest that osteoprotegerin (OPG) and its ligand receptor activator of nuclear factor‐κB ligand (RANKL) constitute a novel cytokine system that can influence the function of both bone and immune cells. The aim of our study was to assess OPG and RANKL concentrations in the early post‐operative period of liver transplantation. Methods: Serum OPG and RANKL levels were measured in 30 patients who underwent liver transplantation at 1, 7 and 14 d post‐operatively. These values were compared with 22 age‐ and sex‐matched healthy controls. Plasma sodium, creatinine, aspartate‐aminotransferase, alanine‐amino transferase, γ‐glutamyl transferase, alkaline phosphatase, bilirubin, albumin, prothrombin time, tacrolimus and cyclosporine levels were measured in each patient. Results: We found a significant increase in OPG levels in the early post‐operative period compared with the control group: day 1 (10.42 pmol/L, range 3.80–17.50 vs. 3.91 pmol/L, range 1.20–6.60; p = 0.0001), day 7 (6.90 pmol/L, range 3.00–15.30 vs. 3.91 pmol/L, range 1.20–6.60; p = 0.0001) and day 14 (5.76 pmol/L, range 2.60–10.70 vs. 3.91 pmol/L, range 1.20–6.60; p = 0.001). Similarly, serum RANKL levels were significantly higher than in the control group in this period, day 1 (0.123 pmol/L, range 0.010–0.420 vs. 0.054 pmol/L, range 0.010–0.300; p = 0.02), day 7 (0.236 pmol/L, range 0.010–0.720 vs. 0.054 pmol/L, range 0.010–0.300; p = 0.0004) and day 14 (0.137 pmol/L, range 0.010–0.520 vs. 0.054 pmol/L, range 0.010–0.300; p = 0.007). No correlation was found between OPG levels and RANKL, ischemic times, liver function tests, albumin, sodium or creatinine concentrations and tacrolimus or cyclosporine levels. Conclusions: A significant amount of OPG and RANKL is released in the early post‐transplant period of liver transplantation. This might be explained by an activation of the immune system caused by the allograft. Therefore, the RANKL/OPG system may be involved in the pathophysiological evolution of transplantation osteoporosis.     

Expressions of intestinal NF-kappaB, TNF-alpha, and IL-6 following traumatic brain injury in rats

BACKGROUND: NF-kappaB regulates a large number of genes involved in the inflammatory response to critical illness, but it is not well known if and how NF-kappaB is activated in the gut following traumatic brain injury (TBI) and what is the role of cytokine-mediated inflammation in the pathogenesis of acute gut mucosal injury. MATERIALS AND METHODS: Male Wistar rats were randomly divided into control and TBI groups, each of which was subgrouped at hours 3, 12, 24, and 72 and on day 7. Parietal brain contusion was produced by a free-falling weight on the exposed dura of the right parietal lobe. NF-kappaB binding activity in jejunal tissue was measured using EMSA and the concentrations of TNF-alpha and IL-6 were detected using ELISA. RESULTS: NF-kappaB binding activity in the jejunum was significantly increased at 3 h following TBI, was maximal at 72 h, and remained elevated by 7 days postinjury. TNF-alpha and IL-6 concentrations were also significantly increased by 3 h postinjury, but peaked at 24 h and remained elevated on Day 7 postinjury. CONCLUSIONS: TBI induced a rapid and persistent up-regulation of NF-kappaB and proinflammatory cytokines in the gut, which may play an important role in the pathogenesis of acute gut mucosal injury mediated by inflammation.     

Targeting the receptor activator of nuclear factor-kappaB (RANK) ligand in prostate cancer bone metastases

Newly formed bone in the typically osteoblastic bone metastases from prostate cancer shows characteristics of woven bone, e.g. marked defects in mineralization and microstructure. Adding to the reduced mechanical strength of prostate cancer bone metastasis is an increasingly recognized osteolytic component. The existence of osteoclasts in osteoblastic bone metastases and concomitant increases in urine or serum markers for bone resorption are reported in affected patients. Pathologically increased osteoclastic bone resorption is a key mediator of the clinical complications from bone metastases, among them fractures, spinal cord compression and bone pain. The receptor activator of nuclear factor (NF)-kappaB ligand (RANKL) pathway has been identified as the main driving force for osteoclastogenesis and resulting bone resorption. Emerging data indicate that bone marrow-derived RANKL might also constitute a chemoattractant factor for RANK-expressing tumour cells that is likely to contribute to the pathogenesis of bone metastases, including those arising from prostate cancer. Cumulative evidence supports RANKL inhibition as a therapeutic goal for the treatment and prevention of bone metastases from prostate cancer.     

Neuropsychological function in patients with increased serum levels of protein S-100 after minor head injury

Summary Protein S-100 is a calcium binding protein, synthetized in astroglial cells in all parts of the central nervous system (CNS). We have previously reported high serum levels of protein S-100 in patients after minor head injury (MHI). A battery of conventional and computerized neuropsychological measures was administered to two groups of MHI patients. Neuropsychological outcome at 12 months postinjury was examined in a group of 7 patients with increased serum levels of protein S-100 after MHI and 7 age- and sex-matched controls without detectable S-100 in serum after MHI. Our results demonstrate no overall cognitive dysfunction in either of the two groups. Our findings indicate specific dysfunction on measures of reaction time, attention and speed of information processing for the S-100 group. Posttraumatic depression does not explain the neuropsychological differences between the groups. These findings support that increased serum levels of protein S-100 may be of predictive and prognostic value for longlasting neurocognitive abnormalities after minor head injury. Presence of S-100 in serum may indicate the presence of diffuse brain damage. Our results suggest that information processing measures in computerized neuropsychological assessment are more sensitive for detecting small signs of neurocognitive abnormalities after MHI than conventional test batteries.     

Prostate-specific antigen stimulates osteoprotegerin production and inhibits receptor activator of nuclear factor-kappaB ligand expression by human osteoblasts

BACKGROUND: Prostate cancer cells produce a large amount of prostate-specific antigen (PSA), which is widely used as a marker for this cancer. Even though it is widely used in the diagnosis of prostate cancer, many aspects of the pathophysiologic role of PSA in bone metastasis remain obscure. The receptor activator of nuclear factor-kappaB ligand (RANKL) is essential for the activation of osteoclasts, while osteoprotegerin (OPG) neutralizes the action of RANKL. Various substances that act on bone have been shown to modulate the production of RANKL and OPG by osteoblasts. METHODS: In this study, we investigated the effect of PSA on the expression of OPG and RANKL mRNA and on protein production in human osteoblast-like cells. RESULTS: After addition of PSA and culture for 72 h, OPG mRNA expression and protein secretion by MG-63 and SaOS-2 cells showed a concentration-dependent increase. When osteoblasts were incubated with PSA (100 ng/ml), OPG mRNA expression and protein secretion increased with the passage of time. alpha1 -antichymotrypsin (ACT), which inactivates the serine protease activity of PSA, inhibited the increase of OPG mRNA expression and protein production in response to PSA, and this effect of PSA was also inhibited by anti-transforming growth factor-beta antibody. CONCLUSIONS: Based on our findings, PSA acts on human osteoblast-like cells via its own serine protease activity and promotes osteoblast differentiation. In addition, PSA stimulates OPG production and inhibits RANKL expression of osteoblasts, and inhibits bone resorption by osteoclasts, suggesting that it contributes to the characteristic osteoblastic features of bone metastases of prostate cancer.     

颅脑损伤后脑脊液TNF-α、IL-6的动态变化

目的探讨颅脑损伤后51例患者伤后不同时期脑脊液中IL-6、TNF—α的变化。方法检测51例急性颅脑损伤患者脑脊液IL-6、TNF—α含量的变化，统计学方法处理分析急性颅脑损伤分级脑脊液中IL-6、TNF—α含量之间的关系以及其变化趋势。结果：与正常对照组相比，颅脑损伤患者伤后第1、3、7、14天脑脊液IL-6均有明显增高（P〈0．01）。重型组明显高于轻、中型组；在轻型组IL-6早期升高，后期下降迅速；中型组于14天接近正常，重型组下降较慢，于14天时仍处于较高水平。本研究发现TNF—α在颅脑损伤后的早期在脑脊液中升高，随即迅速回落至基础值；没有产生与脑水肿高峰期相吻合的第二个峰值或持续升高。结论对于临床诊断和严重程度判断以及指导不同颅脑损伤在某时段遏制炎症，提高颅脑损伤救治水平监测颅脑损伤后患者脑脊液中IL-6具有十分重要的意义．而监测TNF—α水平价值不如IL-6。     

急性胰腺炎时IL-6、TNF-α的变化及大黄干预的研究

目的:探讨大黄对大鼠急性胰腺炎治疗作用的机理。方法:通过胰胆管逆行注射5 %牛磺胆酸钠制作大鼠急性胰腺炎模型,检测血清TNF -α、IL - 6水平及大黄治疗后的变化。结果:大黄治疗后AP大鼠血清TNF -α、IL - 6明显降低。结论:大黄通过降低血清TNF -α、IL - 6水平,可以对实验性急性胰腺炎的治疗产生一定的有益帮助     

Changes in levels of serum IL-8 in burned patients

Interleukin (IL)-8 levels in serial serum samples of 10 burned patients were analysed. The total body surface areas (TBSAs) of the burn injury ranged 30 to 85 per cent. Of these ten patients, five recovered and the other five, who were septic, died. On admission at about 5–13 h postburn, one of the five survivors and two of the non-survivors had serum IL-8 levels higher than 18.1 pg/ml, which is the detection limit of the IL-8 assay kit. The serum IL-8 values of six healthy laboratory personnel included in the present study were all less than 18.1 pg/ml. Afterwards, an initial peak serum IL-8 response was detected within 2–4.5 days postburn. Significant differences in the peak serum IL-8 levels were not found between patients with TBSAs of greater or less than 50 per cent and patients who survived or expired from burn injury. In the survivors, serum IL-8 remained low, whereas IL-8 increased markedly, starting at about one week postburn in four of the five non-survivors with confirmed sepsis. Significant differences in the maximum serum IL-8 levels were detected between patients who recovered vs. those who died from the thermal injury. In conclusion, the results showed that there was an increase in serum IL-8 postburn. Serum IL-8 was significantly higher in the septic patients, who all died. This cytokine may play a significant role in the pathophysiology of sepsis in burned patients.     

Circulating levels of interleukin-6 and its soluble receptor in patients with head injury and fracture

There is evidence that fractures heal more rapidly in patients with head injury. We measured the circulating level of interleukin-6 (IL-6) and its soluble receptor (sIL-6R) and soluble glycoprotein 130 (sgp130) in serum from patients who had sustained a head injury with and without fracture and compared these with levels found in control subjects. Within 12 hours of injury the serum level of IL-6 was significantly higher in patients with head injury and fracture compared with the control group. Levels of IL-6 were also significantly higher in patients with head injury and fracture compared with fracture only. While there was no significant difference in circulating levels of sIL-6R in the initial samples they were increased one week after surgery in patients with head injury and fracture and with head injury only. In addition, reduced levels of sgp130 in patients with head injury with and without fracture indicated a possible reduction of the inhibitory effect of this protein on the activity of IL-6. Our study suggests that IL-6 may be involved in altered healing of a fracture after head injury.     

Expression of osteoprotegerin, receptor activator of NF-kappaB ligand (osteoprotegerin ligand) and related proinflammatory cytokines during fracture healing.

Fracture healing is a unique biological process regulated by a complex array of signaling molecules and proinflammatory cytokines. Recent evidence for the role of tumor necrosis family members in the coupling of cellular functions during skeletal homeostasis suggests that they also may be involved in the regulation of skeletal repair. The expression of a number of cytokines and receptors that are of functional importance to bone remodeling (osteoprotegerin [OPG], macrophage colony-stimulating factor [M-CSF], and osteoprotegerin ligand [receptor activator of NF-kappaB ligand (RANKL)]), as well as inflammation (tumor necrosis factor alpha [TNF-alpha] and its receptors, and interleukin-1alpha [IL-1alpha] and -beta and their receptors) were analyzed over a 28-day period after the generation of simple transverse fractures in mouse tibias. OPG was expressed constitutively in unfractured bones and elevated levels of expression were detected throughout the repair process. It showed two distinct peaks of expression: the first occurring within 24 h after fracture and the second at the time of peak cartilage formation on day 7. In contrast, the expression of RANKL was nearly undetectable in unfractured bones but strongly induced throughout the period of fracture healing. The peak in expression of RANKL did not correlate with that of OPG, because maximal levels of expression were seen on day 3 and day 14, when OPG levels were decreasing. M-CSF expression followed the temporal profile of RANKL but was expressed at relatively high basal levels in unfractured bones. TNF-alpha, lymphotoxin-beta (LT-beta), IL-1alpha, and IL-1beta showed peaks in expression within the first 24 h after fracture, depressed levels during the period of cartilage formation, and increased levels of expression on day 21 and day 28 when bone remodeling was initiated. Both TNF-alpha receptors (p55 and p75) and the IL-1RII receptor showed identical patterns of expression to their ligands, while the IL-1R1 was expressed only during the initial period of inflammation on day 1 and day 3 postfracture. Both TNF-alpha and IL-1alpha expression were localized primarily in macrophages and inflammatory cells during the early periods of inflammation and seen in mesenchymal and osteoblastic cells later during healing. TNF-alpha expression also was detected at very high levels in hypertrophic chondrocytes. These data imply that the expression profiles for OPG, RANKL, and M-CSF are tightly coupled during fracture healing and involved in the regulation of both endochondral resorption and bone remodeling. TNF-alpha and IL-1 are expressed at both very early and late phases in the repair process, which suggests that these cytokines are important in the initiation of the repair process and play important functional roles in intramembraneous bone formation and trabecular bone remodeling.     

人成骨样细胞和前破骨样细胞ER亚型、OPG/RANKL/RANK系统、IL-6及其受体以及破骨细胞标志基因表达的研究

目的分析比较两种人成骨样细胞系(U2-OS、MG-63)和两种人前破骨样细胞系(U937、HL-60)ER亚型、OPG/RANKL/RANK系统、IL-6及其受体以及破骨细胞标志基因表达的差异,为今后研究雌激素与IL-6等细胞因子在骨组织中相互关系提供适宜的细胞模型。方法RT-PCR和免疫印迹法检测ERα、ERβ的表达,ELISA法测定IL-6的分泌,OPG、RANKL、RANK及IL-6受体的表达采用免疫印迹法进行检测,TRAP、MMP-9则采用RT-PCR技术进行分析。结果(1)转录水平及蛋白水平证实四种细胞均表达ERα和ERβ;(2)四种细胞不同程度地表达OPG和RANKL,而RANK则仅见于U937细胞表达;(3)四种细胞均表达IL-6受体(IL-6Rα、gp130),除U2-OS细胞外其余三种细胞均组成型分泌IL-6;(4)破骨细胞标志基因TRAP在两种人前破骨样细胞U937、HL-60中均表达,且表达水平相近;而MMP-9仅在U937细胞中弱表达;两种人成骨样细胞U2-OS、MG-63未见有这两种基因表达。结论筛选出用于研究雌激素与IL-6等细胞因子在骨组织中相互关系的细胞模型,同时也为今后深入阐明骨靶向和其他新型抗骨质疏松雌激素的分子机制研究奠定基础。     

丹红注射液治疗急性脑梗死对神经功能及IL-6、IL-10水平的影响

目的：探讨丹红注射液治疗急性脑梗死患者对神经功能及血清IL-6、IL-10的影响。方法回顾性分析2013年5月～2014年3月我院收治的急性脑梗死患者105例（轻型34例,中型60例,重型11例）,选取健康研究对象50例。按照治疗方法将患者分为两组,对照组52例采用常规治疗,观察组53例在对照组的基础上采用丹红注射液治疗。结果轻型、中型及重型急性脑梗死患者的IL-6、IL-10水平与健康对照组比较明显较高,三组的IL-6、IL-10水平随着神经功能缺损程度评分越高而越高,三组间差异有统计学意义（P＜0.05）。观察组7d、14d的IL-6、IL-10水平及30d的神经功能缺损程度评分与对照组比较明显较低,两组差异有统计学意义（P＜0.05）。结论丹红注射液治疗急性脑梗死患者疗效显著,能使患者血清IL-6、IL-10水平快速降低及神经功能的改善。     

Proteolytic enzyme activity in patients with severe head injury and the effect of a proteinase inhibitor

Summary A study was performed to detect the inhibitory effect of intravenously administered aprotinin (Trasylol^®) on brain and CSF protease activity in 25 patients with severe head injury. The data presented include measurements of CSF protease activity, alpha-1-antitrypsin, alpha-2-macroglobulin, haptoglobulin, polyacrylamidgel-electrophoresis pattern, total protein and hemoglobin content. The results indicate that increased protease activity is present and that this induces autolytic processes which can be inhibited by aprotinin treatment. The survival rate was higher after aprotinin treatment. Total CSF protein content was significantly higher in nonsurvivors than in survivors.Dedicated to Professor Dr. F. Spath on occasion of his 80th birthday     

血液透析患者血管钙化、骨密度与骨保护素及其配体的相互关系

目的 研究维持性血液透析（MHD）患者血管钙化、骨密度与血清骨保护素（OPG）及其配体sRANKL的相互关系。 方法 采用酶联免疫吸附法测定血清OPG、sRANKL水平；X线平片检测腹主动脉、股动脉及桡动脉部位血管钙化，计算血管钙化积分；双能X线骨密度仪测定腰椎及股骨骨密度。对各参数的相互关系进行统计分析。 结果 （1）39例MHD患者中25例（64.1%）在不同部位有不同程度的血管钙化，其中轻度钙化16例（41.0%），中重度钙化9例（23.1%）。中重度钙化者血清OPG水平、OPG/sRANKL比值显著高于轻度钙化者[（342.50±171.53） ng/L比（206.21±137.88） ng/L，t = -2.253，P = 0.025；454.65±455.63比135.31±136.81，t = 59，P = 0.035]，而sRANKL水平差异无统计学意义[（0.10±0.08） pmol/L比(0.12±0.08) pmol/L，t = 0.534，P > 0.05]。多元线性回归分析显示 OPG/sRANKL是血管钙化评分的独立影响因素。（2）与骨量正常者比较，骨量异常者血清OPG水平升高[（249.05±137.66） ng/L比（226.67±170.12） ng/L]，sRANKL水平降低[（0.11±0.08） pmol/L比（0.12±0.02） pmol/L]，OPG/sRANKL比值升高（202.31±219.24比148.08±210.10），但差异均无统计学意义。多元线性回归分析显示OPG/sRANKL是腰椎T值的独立影响因素。（3）多元线性回归分析显示血管钙化评分是腰椎和股骨T值的独立影响因素。 结论 MHD患者血管钙化程度是腰椎及股骨骨密度的独立影响因素， OPG/sRANKL可能在血管钙化和骨密度的关系中起了纽带作用。     

血管内皮生长因子在骨折伴颅脑损伤患者血清中的表达

目的探讨骨折伴颅脑损伤患者血清中血管内皮生长因子(VEGF)与骨折愈合的关系。方法收集我院2008年9月至2010年9月期间收治的骨折患者40例,分为骨折伴颅脑损伤组及单纯骨折组,每组各20例。采用双抗体夹心酶联免疫吸附试验(ELISA)测定所有患者血清中VEGF水平,并摄X线片观察骨痂出现及骨折愈合情况,分析VEGF水平变化及骨折愈合过程。结果骨折伴颅脑损伤患者血清中VEGF含量在7d左右出现高峰期(519±44ng/ml),维持较高水平至42d,以后逐渐下降;单纯骨折患者血清中VEGF含量在14d左右为高峰期(438±28ng/ml),28d左右开始逐渐下降。骨折伴颅脑损伤患者骨痂形成时间(8d±0.3d)明显早于单纯骨折患者(18d±0.1d),P<0.01;骨折伴颅腑损伤患者骨折愈合时间(42d±0.4d)明显早于单纯骨折患者(84d±0.2d),P<0.01。结论骨折伴颅脑损伤患者血清中VEGF含量明显高于单纯骨折患者,骨痂形成及骨折愈合时间均比单纯骨折患者所需时间短。颅脑损伤患者VEGF升高可能是骨折愈合加快的原因之一。