小鼠骨髓源性树突状细胞体外诱导培养方法的比较研究

目的探讨最佳体外诱导培养小鼠成熟树突状细胞（dendritic cells,DC）的方法。方法分离、纯化6周龄C57BL／6小鼠骨髓单核细胞,以含10％胎牛血清、20ng／ml重组小鼠粒细胞-巨噬细胞集落刺激因子（GM—CSF）和10ng／ml重组小鼠白细胞介素-4（IL-4）的RPMI-1640培养基培养7d,然后将细胞分成对照未刺激组、肿瘤坏死因子-α（TNF-α）刺激组和TNF-α＋脂多糖（lipopolysaccharides,LPS）刺激组。继续培养48h后,观察各组细胞形态,检测IL-12、IL-6浓度及细胞表面标志CD11c、CD80、CD86和MHC II。结果培养9d后,两刺激组培养的细胞经相差显微镜观察有DC生长。TNF—α刺激组细胞培养上清液中IL-6、IL-12含量显著高于对照组（P〈0．01）,但显著低于TNF—α＋LPS刺激组（P〈0．05）。3组均高表达CD11c,各组间无显著差异;而CD80、CD86和MHC II表达阳性率TNF-α刺激组显著高于对照组（P〈0．01）,TNF-α＋LPS刺激组显著高于单纯TNF—α刺激组（P〈0．05）。结论联合使用TNF-α与LPS刺激可使DC成熟度提高,分泌IL-6、IL-12增加。     

Peritoneal cytokines as early markers of peritonitis following surgery for colorectal carcinoma: a prospective study

This study was to investigate if measurement of peritoneal cytokines is valuable for an early diagnosis of peritonitis following colorectal surgery. One hundred consecutive patients who were to undergo elective resection for carcinoma of the sigmoid colon or the rectum were investigated. Abdominal exudate was obtained from a drainage tube daily after surgery for measuring interleukin (IL)-1β, IL-6 and tumour necrosis factor (TNF)-α. The relationship between peritoneal cytokine levels during the first 3 days after surgery and the development of peritonitis was investigated. Eight patients developed postoperative peritonitis due to anastomotic leakage and pelvic abscess, which was diagnosed on postoperative days 5-8. Peritoneal cytokine levels on postoperative days 1 and 2 were not significantly different between the 8 patients who developed peritonitis and 92 patients who did not: day 1, IL-1βP=0.32, IL-6 P=0.45, TNF-αP=0.85; day 2, IL-1βP=0.26, IL-6 P=0.68, TNF-αP=0.22. In contrast, the cytokine levels on day 3 were significantly higher in patients who developed peritonitis as compared with patients who did not: IL-1βP=0.008, IL-6 P<0.0001, TNF-αP=0.0001. The cytokines significantly increased during the first 3 days in patients who developed peritonitis: IL-1βP=0.049, IL-6 P=0.03, TNF-αP=0.01, while significantly decreased in patients who did not: IL-1βP<0.0001, IL-6 P<0.0001, TNF-αP<0.0001. The outcomes of this investigation showed that the rise in peritoneal IL-1β, IL-6 and TNF-α levels may be an additional early diagnostic predictor of intraabdominal complications following colorectal surgery.     

Effects of fish oil on lymphocyte proliferation,cytokine production and intracellular signalling in weanling pigs

It has been widely documented that fish oil attenuates inflammatory responses partially via down-regulation of T-lymphocyte function. To determine the anti-inflammatory role of fish oil in weanling pigs, we investigated the effects of fish oil and its functional constituents on peripheral blood lymphocyte proliferation, cytokine production and subsequent intracellular signalling in inflammatory-challenged weanling pigs and in in vitro cultured lymphocytes. Fish oil (7%) or corn oil (7%) was supplemented to 72 crossbred pigs (7.6 ± 0.3 kg BW and 28 ± 3 days of age) in a 2 × 2 factorial experiment that included an Escherichia coli lipopolysaccharide (LPS) challenge (challenged or not challenged). On day 14 and 28 of the experiment, 200 μg/kg BW of LPS or an equivalent amount of sterile saline was administered to the pigs by intraperitoneal injection. Blood samples were collected on days 15 and 29 to determine peripheral blood lymphocyte proliferation, interleukin-1β (IL-1β) and interleukin-2 (IL-2) production. The results showed that inflammatory challenge decreased average daily gain (P < 0.05) and average daily feed intake (P < 0.05) during days 15 - 28. Fish oil supplementation had no effect on growth performance. Inflammatory challenge increased lymphocyte proliferative response to concanavalin A (Con A) (P < 0.05) following each challenge. Fish oil tended to suppress (P < 0.1) the proliferation following the first challenge. Similarly, fish oil tended to reduce IL-1β production (P < 0.1) following the second challenge and IL-2 (P < 0.1) production following the first challenge in both challenged and unchallenged pigs compared with corn oil. In parallel in vitro experiments, peripheral blood lymphocytes of weanling pigs were incubated with various concentrations of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) or linoleic acid (LA) (0, 20, 40, 60, 80, 100 μg/ml). EPA, DHA and high levels of LA predominantly suppressed IL-1β (P < 0.05), IL-2 (P < 0.05) production and subsequent lymphocyte proliferation (P < 0.05). Low levels of LA increased (P < 0.05) IL-2 production. Compared with LA, EPA resulted in a stronger inhibition of lymphocyte proliferation (P < 0.05) and IL-2 (P < 0.01), and DHA resulted in a stronger inhibition of IL-1β (P < 0.05) and IL-2 (P < 0.01). To elucidate the mechanism(s) by which fish oil and its functional constituents suppressed lymphocyte function, the kinetics of intracellular [Ca^{2 +}]_i and protein kinase C activity were determined in in vitro experiments. EPA, DHA and LA exerted very similar dose-dependent stimulatory effects on intracellular Ca^{2 +}. EPA and DHA inhibited protein kinase C activity (P < 0.05), while LA had no significant effect (P > 0.05). These results suggest that fish oil and its functional constituents (EPA and DHA) exerted an anti-inflammatory effect by down-regulation of lymphocyte activation in weanling pigs, possibly by manipulation of intracellular signalling.     

Requirement of Rab21 in LPS-induced TLR4 signaling and pro-inflammatory responses in macrophages and monocytes

Lipopolysaccharide (LPS) induces macrophage/monocyte activation and pro-inflammatory cytokines production by activating Toll-like receptor 4 (TLR-4) signaling. Rab GTPase 21 (Rab21) is a member of the Rab GTPase subfamily. In the present study, we show that LPS induced TLR4 and Rab21 association and endosomal translocation in murine bone marrow–derived macrophages (BMDMs) and primary human peripheral blood mononuclear cells (PBMCs). In BMDMs, shRNA-mediated stable knockdown of Rab21 inhibited LPS-induced expression and production of pro-inflammatory cytokines (IL-1β, IL-6 and TNF-α). Conversely, forced overexpression of Rab21 by an adenovirus construct potentiated LPS-induced IL-1β, IL-6 and TNF-α production in BMDMs. Further studies show that LPS-induced TLR4 endosomal traffic and downstream c-Jun and NFκB (nuclear factor-kappa B) activation were significantly inhibited by Rab21 shRNA, but intensified with Rab21 overexpression in BMDMs. Finally, in the primary human PBMCs, siRNA-induced knockdown of Rab21 significantly inhibited LPS-induced IL-1β, IL-6 and TNF-α production. Taken together, we suggest that Rab21 regulates LPS-induced pro-inflammatory responses by promoting TLR4 endosomal traffic and downstream signaling activation.     

Effects of dietary nucleotide supplementation on growth performance and hormonal and immune responses of piglets

The effects of dietary nucleotide supplementation from 9 days of age until the end of post-weaning on piglets hormonal and immune responses and on growth performance were investigated. During lactation (days 9 to 21) and post-weaning (days 22 to 55) 10 [HBI Fomeva11 × (Large White × Landrace)] litters (n = 108 piglets) had ad libitum access to two standard diets, both supplemented with 0% (T0 group) or 0.1% (T1 group) of yeast extract nucleotides. BW of piglets at days 21 (P < 0.10), 35 and 55 (P < 0.05) was greater in T1 compared with T0. Feed intake was not different between groups (P > 0.05). Cortisol content was lower in T1 than in T0 at days 28 and 35 (P < 0.05), whereas growth hormone was lower at day 35 (P < 0.05). Levels of IGF-1 were similar across groups (P > 0.05). Nucleotide-supplemented diets increased lymphocyte subpopulation CD4-CD8+high at days 21 and 35 (P < 0.05), whereas CD4+CD8- cells were higher in T1 than in T0 at day 21 (P < 0.05). Peripheral blood mononuclear cells cytokine expression was influenced by dietary nucleotide supplementation. At weaning, interleukin (IL)-6 and IL-1β expression was lower (P < 0.05) in T1 compared with T0, whereas the expression of interferon (IFN)-γ and IL-10 was higher (P < 0.05). At day 28, piglets in T1 showed higher values of tumor necrosis factor (TNF)-α expression than T0 and lower values of IL-10 expression (P < 0.05). Dietary nucleotide supplementation had a suppressive effect on IL-6 and IL-10 expression (P < 0.05) at day 35. On the contrary, the expression of IFN-γ, TNF-α and IL-1β was enhanced (P < 0.05). In conclusion, these results suggest that starting a dietary nucleotide supplementation before weaning can improve the adaptive capabilities of weaned piglets to the stressors, enhancing the growth performance.     

Enhancement of the acute phase response to a lipopolysaccharide challenge in steers supplemented with chromium

The study examined the effect of chromium supplementation on the response of steers to an LPS challenge. Steers received a premix that added 0 (control; n?=?10) or 0.2 mg/kg of chromium (n?=?10) to the total diet on a dry matter basis for 56 d. Steers were fitted with jugular catheters and rectal temperature (RT) recording devices on d 52. Blood samples were collected and sickness behavior scores assigned to each steer relative to an LPS challenge (0.5 μg/kg) on d 55. Pre-LPS RT were greater in chromium-supplemented than in control steers. Post-LPS RT increased in both treatments, with control steers producing a greater change in RT than chromium-supplemented steers. Sickness behavior scores were greater in control than in chromium-supplemented steers post-LPS (P?=?0.03). Cortisol concentrations did not differ between treatments pre-LPS. Post-LPS cortisol concentrations increased but did not differ due to treatment. Concentrations of IL-4 increased post-LPS but were not affected by treatment pre- or post-LPS. Treatment did not affect pre-LPS TNF-α or IFN-γ. Post-LPS TNF-α and IFN-γ increased in both treatments, with chromium-supplemented steers producing greater TNF-α (P?=?0.005) and IFN-γ (P?=?0.004) than control steers. Pre-LPS IL-6 was greater (P?=?0.027) in chromium-supplemented steers than in control steers. Post-LPS IL-6 increased in both treatments and was greater (P?相似文献   

Inhibition of p38 MAPK during cellular activation modulate gene expression of head kidney leukocytes isolated from Atlantic salmon (Salmo salar) fed soy bean oil or fish oil based diets

Head kidney leukocytes isolated from Atlantic salmon fed either a diet based on fish oil (FO) or soy bean oil (VO) were used in order to evaluate if different lipid sources could contribute to cellular activation of the salmon innate immune system. A specific inhibitor of p38 MAPK, SB202190, was used to investigate the effect of lipopolysaccharide (LPS) signalling in the head kidney leukocytes. The results show that LPS up regulate IL-1β, TNF-α, Cox2 expression in leukocytes isolated from fish fed either diet. The p38 MAPK inhibitor, SB202190, reduced the LPS induced expression of these genes in both dietary groups. In LPS stimulated leukocytes isolated from VO fed fish, SB202190 showed a clear dose dependent inhibitory effect on IL-1β, TNF-α and Cox2 expression. This effect was also observed for Cox2 in leukocytes isolated from FO fed fish. Furthermore, there was a stronger mean induction of Cox2 in LPS stimulated leucocytes isolated from the VO-group compared to LPS stimulated leukocytes isolated from the FO-group. In both dietary groups, LPS stimulation of salmon head kidney leukocytes increased the induction of CD83, a dendrite cell marker, while the inhibitor reduced CD83 expression in the VO fed fish only. The inhibitor also clearly reduced hsp27 expression in VO fed fish. Indicating a p38 MAPK feedback loop, LPS significantly inhibited the expression of p38MAPK itself in both diets, while SB202190 increased p38MAPK expression especially in the VO diet group. hsp70 expression was not affected by any treatment or feed composition. There were also differences in p38MAPK protein phosphorylation comparing treatment groups but no obvious difference comparing the two dietary groups. The results indicate that dietary fatty acids have the ability to modify signalling through p38 MAPK which may have consequences for the fish's ability to handle infections and stress. Signalling through p38MAPK is ligand dependent and affects gene and protein expression differently.     

骨髓间充质干细胞对肾移植受者T淋巴细胞分化和miRNA-155表达的影响

目的探讨骨髓间充质干细胞（MSCs）对肾移植受者T淋巴细胞分化和miRNA-155表达的影响。 方法选取2013年1月至2017年12月于福州总医院接受MSCs诱导+同种异体肾移植术的受者20例（MSCs组）,对照组为同期配对的异体肾移植受者20例。两组患者术后免疫抑制方案均为霉酚酸酯+他克莫司+强的松。两组患者分别于移植术前、术后第15天抽取静脉血,流式细胞仪检测外周血CD4⁺ CD25⁺ FoxP3⁺ Treg细胞/?CD4⁺细胞亚群比值;ELISA检测白细胞介素2（IL-2）、肿瘤坏死因子α（TNF-α）和白细胞介素10（IL-10）浓度;免疫磁珠分选外周血T淋巴细胞后,Rea1-time PCR法检测外周血T淋巴细胞中miRNA-?155的表达。两组间均数比较采用独立t检验,治疗前后均数比较采用配对t检验。 结果移植前两组肾移植受者外周血CD4⁺ CD25⁺ FoxP3⁺ Treg细胞/?CD4⁺细胞比值、IL-2、IL-?10、TNF-α、T淋巴细胞miRNA-155表达水平差异均无统计学意义（P均> 0.05）;术后第15天,与对照组相比,MSCs组外周血CD4⁺ CD25⁺ FoxP3⁺ Treg细胞/?CD4⁺细胞亚群比值（30.44﹪?± 4.23﹪? vs 26.06﹪?±4.77﹪,t = 2.365,P = 0.042）、IL-10水平（20.35?ng/?L?±?5.10?ng/?L? vs 16.63?ng/?L±6.26?ng/?L,t = 2.062,P?=?0.046）上升,而IL-2（27.47ng/?L±4.30 ng/?L vs 31.40?ng/?L±5.33 ng/L,t = 2.252,P = 0.015）、TNF-α（41.52?ng/?L±8.32?ng/L vs 46.67?ng/?L±6.71?ng/L,t = 2.157,P = 0.037）和T淋巴细胞miRNA-155表达水平（1.61±0.31 vs 1.89±0.15,t = 3.688,P?= 0.001）则降低。 结论MSCs能够升高肾移植受者外周血CD4⁺ CD25⁺ FoxP3⁺ Treg细胞/?CD4⁺细胞比值和IL-10水平,降低IL-2、TNF-α和T淋巴细胞miRNA-155表达水平,与MSCs的免疫耐受诱导有关。     

Preventive oral supplementation with glutamine and arginine has beneficial effects on the intestinal mucosa and inflammatory cytokines in endotoxemic rats

The objective of this study was to evaluate the effect of oral supplementation with a combination of arginine and glutamine on the intestinal mucosa and inflammatory cytokines of lipopolysaccharide (LPS)-induced adult rats. Fifty Sprague-Dawley rats (average weight of 185 ± 15 g) were randomly divided into five groups: control group A (CA) and control group B (CB), both orally supplemented with 0.9% saline; group Arg, supplemented with 300 mg/kg day(-1) arginine; group Gln, supplemented with 300 mg/kg day(-1) glutamine; group AG, supplemented with 150 mg/kg day(-1) arginine and 150 mg/kg day(-1) glutamine. The experiment lasted for 2 weeks. Food intake and body weight were measured during the experiment. At 10.00 h of day 15, animals were injected with 4 mg/kg LPS (group CB, Arg, Gln, and AG) or sterile saline (group CA) after supplementation. Then at 14.00 h, all animals were killed and blood and tissue collected. The results showed that compared with group CB, arginine concentration tended to be increased (P > 0.05) in group Arg and AG, while there was no significant difference in glutamine concentration among the groups challenged with LPS. Oral supplementation with arginine or/and glutamine mitigated morphology impairment (lower villus height, P < 0.05) in the jejunum and ileum induced by LPS challenge. LPS administration resulted in a significant increase in TNF-α, IL-1β, IL-6 and IL-10 mRNA abundance. Arginine only significantly decreased TNF-α mRNA abundance in the ileum, while glutamine significantly decreased both TNF-α and IL-10 mRNA in the ileum. A combination of arginine and glutamine significantly decreased TNF-α and IL-1β mRNA abundance in both the jejunum and ileum, while they also significantly decreased anti-inflammatory IL-10 in the ileum. These results revealed that an oral supply of combined arginine and glutamine had more favorable effects on the intestinal mucosa and inflammatory cytokines than a supply of arginine or glutamine alone.     

老年急性脑梗塞患者血清TNF-α 和IL-6 水平的变化及其临床意义

目的:观察老年急性脑梗塞(ACI)患者血清TNF-α和IL-6水平的变化并探讨其临床意义.方法:选择老年ACI患者53例,观察以上入选患者入院后第7d、第9d和第11d血清TNF-α和IL-6水平的变化,并与26例健康人对照;同时分析53例老年ACI患者血清TNF-α和IL-6水平与患者脑梗塞面积及神经功能缺损评分的Spearman等级相关性.结果:老年ACI患者血清TNF-α和IL-6水平显著高于健康人(P＜0.01);随着治疗的进展与病情的稳定及恢复,患者血清TNF-α和IL-6水平随之显著下降(P＜0.05);同时患者血清TNF-α和IL-6水平与病变的严重程度呈正相关性(P＜0.05);治疗11d后血清TNF-α和IL-6降低量与病变的严重程度呈正相关性(P＜0.05).结论:在老年ACI患者治疗过程中,应进行血清TNF-α和IL-6水平的动态监测,可提示病变的发生发展并指导临床治疗.     

Effects of feed access after hatch and inclusion of fish oil and medium chain fatty acids in a pre-starter diet on broiler chicken growth performance and humoral immunity

Delayed feed and water access is known to impair growth performance of day old broiler chickens. Although effects of feed access on growth performance and immune function of broilers have been examined before, effects of dietary composition and its potential interaction with feed access are hardly investigated. This experiment aimed to determine whether moment of first feed and water access after hatch and pre-starter composition (0 to 7 days) affect growth rate and humoral immune function in broiler chickens. Direct fed chickens received feed and water directly after placement in the grow-out facility, whilst delayed fed chickens only after 48 h. Direct and delayed fed chickens received a control pre-starter diet, or a diet containing medium chain fatty acids (MCFA) or fish oil. At 21 days, chickens were immunized by injection of sheep red blood cells. The mortality rate depended on an interaction between feed access and pre-starter composition (P=0.014). Chickens with direct feed access fed the control pre-starter diet had a higher risk for mortality than chickens with delayed feed access fed the control pre-starter diet (16.4% v. 4.2%) whereas the other treatment groups were in-between. BW gain and feed intake till 25 days in direct fed chickens were higher compared with delayed fed chickens, whilst gain to feed ratio was lower. Within the direct fed chickens, the control pre-starter diet resulted in the highest BW at 28 days and the MCFA pre-starter diet the lowest (Δ=2.4%), whereas this was opposite for delayed fed chickens (Δ=3.0%; P=0.033). Provision of MCFA resulted in a 4.6% higher BW gain and a higher gain to feed ratio compared with other pre-starter diets, but only during the period it was provided (2 to 7 days). Minor treatment effects were found for humoral immune response by measuring immunoglobulins, agglutination titers, interferon gamma (IFN-γ), and complement activity. Concluding, current inclusion levels of fish oil (5 g/kg) and MCFA (30 g/kg) in the pre-starter diet appear to have limited (carryover) effects on growth and development, as well as on humoral immune function.     

Dietary n-3 polyunsaturated fatty acids promote activation-induced cell death in Th1-polarized murine CD4+ T-cells

Dietary n-3 PUFAs have been shown to attenuate T-cell-mediated inflammation. To investigate whether dietary n-3 PUFAs promote activation-induced cell death (AICD) in CD4+ T-cells induced in vitro to a polarized T-helper1 (Th1) phenotype, C57BL/6 mice were fed diets containing either 5% corn oil (CO; n-6 PUFA control) or 4% fish oil (FO) plus 1% CO (n-3 PUFA) for 2 weeks. Splenic CD4+ T-cells were cultured with alpha-interleukin-4 (alphaIL-4), IL-12, and IL-2 for 2 days and then with recombinant (r) IL-12 and rIL-2 for 3 days in the presence of diet-matched homologous mouse serum (HMS) to prevent loss of cell membrane fatty acids, or with fetal bovine serum. After polarization, Th1 cells were reactivated and analyzed for interferon-gamma and IL-4 by intracellular cytokine staining and for apoptosis by Annexin V/propidium iodide. Dietary FO enhanced Th1 polarization by 49% (P = 0.0001) and AICD by 24% (P = 0.0001) only in cells cultured in the presence of HMS. FO enhancement of Th1 polarization and AICD after culture was associated with the maintenance of eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3) in plasma membrane lipid rafts. In conclusion, n-3 PUFAs enhance the polarization and deletion of proinflammatory Th1 cells, possibly as a result of alterations in membrane microdomain fatty acid composition.     

Effects of β-glucan extracted from Saccharomyces cerevisiae on humoral and cellular immunity in weaned piglets

Abstract

Twenty-four barrows were used to investigate the effects of β-glucan on immune function in weaned piglets. Pigs (8.09 ± 0.20 kg, 28 d of age) were fed a diet without or with supplemented β-glucan (50 mg/kg feed). All pigs were injected with ovalbumin (OVA) on day 14 to investigate their humoral immune response. On day 28, lymphocytes were isolated from all pigs to determine the effects of β-glucan on cellular immunity of pigs in vitro. Lymphocytes from six pigs of each group were incubated with 16 μg lipopolysaccharide (LPS) per ml culture medium, the remainder with an equivalent volume of culture medium alone. Samples were collected at 0, 3, 6, 12, 18, 24, and 48 h after LPS addition for determination of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-10 (IL-10). On day 31, six pigs of each group were injected with either LPS (25 μg/kg BW) or an equivalent amount of sterile saline. Blood samples were collected at 3 h after LPS injection for analysis of IL-6, TNF-α, and IL-10 in plasma. The results indicated that dietary β-glucan enhanced pig antibody response to OVA only in the first week after injection. In vitro, the increases of IL-6 and TNF-α in culture medium were partially dampened in pigs supplemented with β-glucan when their lymphocytes were incubated with LPS, whereas the increase of IL-10 was potentiated. In vivo, dietary β-glucan attenuated the increase of plasma IL-6 and TNF-α, and enhanced the increase of plasma IL-10 when pigs were challenged with LPS. These results demonstrate that β-glucan can improve the humoral immunity of pigs and modulate cellular immunity of pigs by mitigating the elevation of pro-inflammatory cytokines and enhancing the increase of anti-inflammatory cytokines after an immunological challenge.     

Dendritic cell-derived TNF-α is responsible for development of IL-10-producing CD4 T cells

Immature dendritic cells (DCs) appear to be involved in peripheral immune tolerance via induction of IL-10-producing CD4⁺ T cells. We examined the role of TNF-α in generation of the IL-10-producing CD4⁺ T cells by immature DCs. Immature bone marrow-derived DCs from wild type (WT) or TNF-α^−/− mice were cocultured with CD4⁺ T cells from OVA specific TCR transgenic mice (OT-II) in the presence of OVA_323-339 peptide. The WT DCs efficiently induced the antigen-specific IL-10-producing CD4⁺ T cells, while the ability of the TNF-α^−/− DCs to induce these CD4⁺ T cells was considerably depressed. Addition of exogenous TNF-α recovered the impaired ability of the TNF-α^−/− DCs to induce IL-10-producing T cells. However, no difference in this ability was observed between TNF-α^−/− and WT DCs after their maturation by LPS. Thus, TNF-α appears to be critical for the generation of IL-10-producing CD4⁺ T cells during the antigen presentation by immature DCs.     

Role of Proinflammatory Cytokines on Lipopolysaccharide-Induced Phase Shifts in Locomotor Activity Circadian Rhythm

We previously reported that early night peripheral bacterial lipopolysaccharide (LPS) injection produces phase delays in the circadian rhythm of locomotor activity in mice. We now assess the effects of proinflammatory cytokines on circadian physiology, including their role in LPS-induced phase shifts. First, we investigated whether differential systemic induction of classic proinflammatory cytokines could explain the time-specific behavioral effects of peripheral LPS. Induction levels for plasma interleukin (IL)-1α, IL-1β, IL-6, or tumor necrosis factor (TNF)-α did not differ between animals receiving a LPS challenge in the early day or early night. We next tested the in vivo effects of central proinflammatory cytokines on circadian physiology. We found that intracerebroventricular (i.c.v.) delivery of TNF-α or interleukin IL-1β induced phase delays on wheel-running activity rhythms. Furthermore, we analyzed if these cytokines mediate the LPS-induced phase shifts and found that i.c.v. administration of soluble TNF-α receptor (but not an IL-1β antagonistic) prior to LPS stimulation inhibited the phase delays. Our work suggests that the suprachiasmatic nucleus (SCN) responds to central proinflammatory cytokines in vivo, producing phase shifts in locomotor activity rhythms. Moreover, we show that the LPS-induced phase delays are mediated through the action of TNF-α at the central level, and that systemic induction of proinflammatory cytokines might be necessary, but not sufficient, for this behavioral outcome. (Author correspondence: dgolombek@unq.edu.ar)     

Orally administered LPS enhances head kidney macrophage activation with down-regulation of IL-6 in common carp (Cyprinus carpio)

Immunostimulants represent a promising aquaculture tool for enhancing disease and stress resistance in cultured fish. Moreover, the term and dose for acting immunostimulants is an important thing for fish farmer. This study investigated the immune parameters of common carp after oral administration of LPS (5, 10, 20 μg/kg/days) for 30 and 60 days, which is considered to be the proper time period for acting in aquaculture. Phagocytic and bactericidal activities of head kidney macrophages and serum lysozyme activities were significantly enhanced in LPS-fed carp. Orally administered LPS augmented the expression of interleukin (IL)-1β and TNF-α mRNAs but reduced the expression of IL-6 mRNA in head kidney. Although LPS was detected in the serum and liver after a high-dose (>15 mg/kg) oral administration, it was not detected by administered LPS-specific ELISA after a low-dose (<20 μg/kg) administration. It is speculated that orally administered LPS enhances the eliminating functions of head kidney macrophages with down-regulation of IL-6.     

凝结芽胞杆菌活菌片对非酒精性脂肪性肝炎患者内毒素及细胞因子的影响

目的探讨凝结芽胞杆菌活菌片（商品名：爽舒宝）对非酒精性脂肪性肝炎患者内毒素及细胞因子的影响。方法将41例非酒精性脂肪性肝炎患者随机分为观察组和对照组,观察组在常规治疗的基础上加服凝结芽胞杆菌活菌片,对照组只给予常规治疗,治疗前后测定血内毒素、TNF-α、IL-6指标的变化。结果观察组治疗后各项指标的水平均显著低于对照组,差异有显著性（P〈0.01）。结论凝结芽胞杆菌活菌片通过降低血内毒素、TNF-α、IL-6,对非酒精性脂肪性肝炎患者具有显著的治疗作用。