ObjectiveTo assess the effect of atorvastatin on lipopolysaccharide(LPS)-inducedTNF-α production in RAW264.7 macrophages. MethodsRAW264.7 macrophageswere treated in different LPS concentrations oratdifferent time points with or without atorvastatin. TNF-α level in supernatant was measured. Expressions of TNF-αmRNA and protein and heme oxygenase-1 (HO-1) were detected by ELISA, PCR, and Western blot, respectively. HOactivity was assayed. ResultsLPS significantly increased the TNF-α expression and secretion in a dose- and time-dependent manner. The HO-1activity and HO-1 expression level were significantly higher after atorvastatin treatment than before atorvastatin treatment and attenuated by SB203580 and PD98059 but not by SP600125, suggesting that the ERK and p38 mitogen-activated protein kinase (MAPK) pathways participate inregulating the above-mentioned effects of atorvastatin. Moreover, the HO-1 activity suppressed by SnPP or the HO-1 expression inhibited by siRNA significantly attenuated the effect of atorvastatin onTNF-α expression and production in LPS-stimulated macrophages. ConclusionAtorvastatin can attenuate LPS-induced TNF-α expression and production by activating HO-1 via the ERK and p38 MAPK pathways,suggesting that atorvastatin can be used in treatment of inflammatory diseases such as sepsis, especially in those with atherosclerotic diseases.

Ganoderma lucidum is a traditional Chinese medicine, which has been shown to have both an-ti-oxidative and anti-inlfammatory effects, and noticeably decreases both the infarct area and neuronal apoptosis of the ischemic cortex. This study aimed to investigate the protective effects and mechanisms of pretreatment with ganoderma lucidum (by intragastric administration) in cerebral ischemia/reperfusion injury in rats. Our results showed that pretreatment with ganoder-ma lucidum for 3 and 7 days reduced neuronal loss in the hippocampus, diminished the content of malondialdehyde in the hippocampus and serum, decreased the levels of tumor necrosis fac-tor-αand interleukin-8 in the hippocampus, and increased the activity of superoxide dismutase in the hippocampus and serum. hTese results suggest that pretreatment with ganoderma lucidum was protective against cerebral ischemia/reperfusion injury through its anti-oxidative and an-ti-inlfammatory actions.

Previous studies have shown that vagus nerve stimulation can improve the prognosis of trau-matic brain injury. The aim of this study was to elucidate the mechanism of the neuroprotective effects of vagus nerve stimulation in rabbits with brain explosive injury. Rabbits with brain ex-plosive injury received continuous stimulation (10 V, 5 Hz, 5 ms, 20 minutes) of the right cervical vagus nerve. Tumor necrosis factor-α, interleukin-1βand interleukin-10 concentrations were detected in serum and brain tissues, and water content in brain tissues was measured. Results showed that vagus nerve stimulation could reduce the degree of brain edema, decrease tumor necrosis factor-αand interleukin-1βconcentrations, and increase interleukin-10 concentration after brain explosive injury in rabbits. These data suggest that vagus nerve stimulation may exert neuroprotective effects against explosive injury via regulating the expression of tumor necrosis factor-α, interleukin-1βand interleukin-10 in the serum and brain tissue.

Inlfammation may play a role in postoperative cognitive dysfunction. 5′ Adenosine monophos-phate-activated protein kinase, nuclear factor-kappa B, interleukin-1β, and tumor necrosis factor-αare involved in inlfammation. Therefore, these inlfammatory mediators may be involved in postoperative cognitive dysfunction. Western immunoblot analysis revealed 5′adenosine mo-nophosphate-activated protein kinase and nuclear factor-kappa B in the hippocampus of aged rats were increased 1-7 days after splenectomy. Moreover, interleukin-1βand tumor necrosis fac-tor-αwere upregulated and gradually decreased. Therefore, these inlfammatory mediators may participate in the splenectomy model of postoperative cognitive dysfunction in aged rats.

Alzheimer’s disease is closely associated with disorders of neurogenesis in the brain, and growing evidence supports the involvement of immunological mechanisms in the development of the disease. However, at present, the role of T cells in neuronal regeneration in the brain is unknown. We injected amyloid-beta 1-42 peptide into the hippocampus of six BALB/c wild-type mice and six BALB/c-nude mice with T-cell immunodeifciency to establish an animal model of Alzhei-mer’s disease. A further six mice of each genotype were injected with same volume of normal saline. Immunohistochemistry revealed that the number of regenerated neural progenitor cells in the hippocampus of BALB/c wild-type mice was signiifcantly higher than that in BALB/c-nude mice. Quantitative fluorescence PCR assay showed that the expression levels of peripheral T cell-associated cytokines (interleukin-2, interferon-γ) and hippocampal microglia-related cyto-kines (interleukin-1β, tumor necrosis factor-α) correlated with the number of regenerated neural progenitor cells in the hippocampus. hTese results indicate that T cells promote hippocampal neurogenesis in Alzheimer’s disease and T-cell immunodeifciency restricts neuronal regeneration in the hippocampus. The mechanism underlying the promotion of neuronal regeneration by T cells is mediated by an increased expression of peripheral T cells and central microglial cytokines in Alzheimer’s disease mice. Our ifndings provide an experimental basis for understanding the role of T cells in Alzheimer’s disease.

孕激素在子痫前期患者TLR4-MyD88依赖的信号通路中的作用

目的 研究孕激素在子痫前期(preeclampsia,PE)孕妇Toll样受体4(toll-like receptor4,TLR4)-髓样细胞分化蛋白88(myeloid differentiation factor 88,MyD88)依赖的信号通路中的作用.方法 原代培养子痫前期孕妇外周血单个核细胞(peripheral blood mononuclear cell,PBMC),以不同浓度孕激素(0 mol/L、10-8 mol/L、10-6mol/L、 10-4mol/L)处理,实时定量聚合酶联反应(Real-time polymerase Chain reaction,real-time PCR)检测TLR4 mRNA、MyD88 mRNA、核转录因子-κB (nuclear factor κB,NF-κB) mRNA的表达;免疫印迹法Western blot检测IκB-α蛋白的表达,用凝胶图像处理系统分析各条目标带灰度值酶联免疫吸附试验(Enzyme-linked immunosorbent assay,ELISA)检测细胞上清液中肿瘤坏死因子-α (tumor necrosis factors,TNF-α)及白介素-6(interleukin-6,IL-6)的表达.结果 随着孕酮浓度的增加,各组TLR4 mRNA、MyD88 mRNA及NF-κB mRNA的相对表达量2-△△Ct均逐渐降低,IκB-α蛋白的表达逐渐增加,差异均有统计学意义(均P<0.05);同时,细胞上清液中细胞因子TNF-α及IL-6的表达均逐渐减少,差异均有统计学意义(P<0.05).结论 孕激素可显著抑制TLR4-MyD88依赖的信号传导通路,对子痫前期患者具有保护作用.

Effect of seawater immersion on periosteal function of the soft tissue with open injury in rabbit limbs

Objective To investigate the effect of seawater immersion on periosteal function of the soft tissue with open injury in rabbit limbs.Methods Forty-eight New Zealand rabbits were used in the experiment.The animal model of open soft tissue injury was developed by cutting open the hindlimbs,and then the animals were randomly divided into 3 groups:the simple injury group or the control group ( group A),the injury plus 30 min seawater immersion group (group B),and the injury plus 1 h seawater immersion group (group C).Rabbits were sacrificed following surgery on 0,1,3,7 days,and histopathology and expressions of TNF-α and TGF-β1 were observed and monitored from the collected periosteum.Results (1) Inflammatory response of periosteum resulting from limb surgery plus 30 min seawater immersion was more serious than that of the animals in the control group.The longer the immersion,the more serious the response.(2) Intramembranous bone formation delayed markedly,following 1 h seawater immersion,however,for the animals in group B,delay in intramembranous bone formation was insignificant. (3) Periosteal malpighian layer cells for the animals in group B increased significantly,following injury on 1,3,7 days.Marked differences could be seen,when comparisons were made with animals of group A and group C (P ＜ 0.01 ).Though periosteal malpighian layer cells for the animals in group A increased,when compared with those of the animals in group C,no significant differences could be noted between them ( P ＞ 0.05 ).(4) Expressions of TNF-α were all seen in periosteal osteoprogenitor cells and osteoblasts in the animals of the 3 groups.The expression reached peak on day 1 after surgery,then,decreased gradually,with the expression level increased with the extension of seawater immersion. (5)There were expressions of TGF-β1 in periosteal osteoprogenitor cells and osteoblasts in the animals of the 3 groups and persistent expression of TGF-β1 could be observed on day 0 to day 7,and the expression of TGF-β1 in the animals of group B was obviously stronger than those of the animals in group C and group A.Conclusions Seawater immersion could aggravate periosteal damage,and 1 h seawater immersion could obviously delay and damage the capability of intramembranous bone formation of periosteum, indicating that the longest endurance of periosteum with seawater immersion could be less than 1 hour.

Objective: To study the effects of paclitaxel on macrophage activation. Methods:Mouse macrophages were isolated by peritoneal lavage and cultured in RPMI 1640 medium according to the following groups: paclitaxel (5μmol/L) group, IFN-γ (5U/L) group, paclitaxel (5μmol/L) and IFN-γ (5U/L) combination group, and control group(without paclitaxel and IFNγ) .24 hours later, supematants were collected for nitric oxide(NO) assessment using the Griess reagent, and ttanor necrosis factor-α(TNF-α) assessment using the enzyme linked immunosorbent assay. Antibody-dependent cell-mediated cytotoxicity(ADCC) of the macrophages was assessed using the method of hemoglobin-enzyme release assay (Hb-ERA). Results: Paclitaxel induced the production of higher levels of NO(8.86 ± 1.16μmol/L) and TNF-α(120.2 ± 10.2pg/ml) ,and enhanced the ADCC of macrophages[ (20.61 + 1.13)% ]. The differences were significant compared with the control group[no NO and TNF-α detected,ADCC (15.37 + 1.93)% ](P ＜ 0.01). Paclitaxel and IFN-γ in combination induced the production of higher levels of NO(22.85 ± 0.91μmol/L) and TNF-α(358.6 ± 27 .5pg/ml), and enhanced the ADCC of macrophages[ (42.49 + 3.09) % ]. The differences were significant compared with paclitaxel or IFN-γ[NO 8.09 ± 1.13μmol/L, TNF-α1 24.8 + 9.6pg/ml, ADCC(23.32 ± 2.63) % ] alone (P＜0.01). Conclusion: These findings indicate that paclitaxel can promote NO and TNF-α production,enhance ADCC of macrophages, and induce macrophage activation. The active effects are more significant with paclitaxel and IFN-γcombination.