钩藤中钩藤碱对肿瘤坏死因子-α诱导人星型胶质细胞炎症模型作用机制研究

目的研究钩藤中钩藤碱对肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)诱导人星型胶质细胞(Human Astrocytes,HA)的炎症模型的作用机制.方法:体外培养HA,用TNF-α诱导HA造模,将细胞分为正常组、模型组、钩藤碱组、依达拉奉组.采用噻唑蓝(MTT)法检测钩藤碱对HA增殖能力的影响,用逆转录PCR法(RT-PCR)检测一氧化氮合酶(nitric oxide synthase,NOS)mRNA、白细胞介素-23(interleukin-23,IL-23)mRNA、白细胞介素-6(interleukin-6,IL-6)mRNA的表达,用分光光度法检测一氧化氮(nitric oxide,NO)、超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)的表达,用蛋白免疫印迹法(Western blot)检测p-mTOR,mTOR,p-Stat3和Stat3的蛋白表达.结果与正常组比较,32ng·mL-1的TNF-α对星形胶质细胞有明显抑制作用(P0.001);400μmol·L~(-1)的钩藤碱对星形胶质细胞活力有明显抑制作用(P0.001).与模型组相比,200μmol·L~(-1)的钩藤碱对模型细胞有明显的增殖作用(P0.01);钩藤碱组和依达拉奉组NOS mRNA,NO和SOD都显著上升(P0.001),IL-23mRNA,IL-6mRNA和MDA都显著下降(P0.001);钩藤碱组p-mTOR/mTOR和p-Stat3/Stat3都显著下降(P0.01).结论钩藤有效成分钩藤碱具有抑制TNF-α诱导HA炎症的损伤作用,可能是通过下调mTOR/STAT3信号通路增加氧化相关因子NOS,NO,SOD的表达,减少氧化相关因子MDA以及炎症相关因子IL-23和IL-6的表达.     

IL-1β regulates the mouse Fas ligand expression in corneal endothelial cells

Constitutively expressed Fas ligand (FasL) in several distinct epithelial cell types appears to protect tissues by inducing apoptosis of Fas immune cells during inflammatory reactions. To study the rela-tionship of FasL and inflammation process in cornea,we examined the effects of inflammatory cytokine IL-1β on the FasL production,expression and cytotoxic function in corneal endothelial cells. In this paper,we demonstrate that IL-1β inhibits the FasL production and expression in corneal endothelial cells. The promoter activities of FasL in these cells are reduced by IL-1β in a dose-dependent manner. Finally,we also find that IL-1β block the cytotoxic effects of FasL derived from corneal endothelial cells to the Fas target cells. These data support the view that FasL derived from corneal endothelial cells modulate inflammation within cornea.     

Effect of MTEC1 on the functional expression of TCRaβ+ 3G11− 6C10− CD4+ CD8− thymocyte subset

A murine CD4⁺ thymocyte subset with phenotype of TCRαβ⁺ 3G11⁻ 6C10⁻ CD4⁺ CD8⁻ CD69^+/- HSA^med/lo contains the cells in relatively functional matured status. The functional property of the cells in this subset is characterized by the unique pattern of cytokine production at transitional stage from Th0 to Th2 type with the latter being the dominant type. After being co-cultured with murine thymic medullary epithelial cell line (MTEC1) cells, a murine thymic medullary type epithelial cell line, the TCRαβ⁺ 3G11⁻ 6C10⁻ CD4⁺ CD8⁻ CD69^+/- HSA^med/lo thymocytes, has exhibited significantly higher levels of proliferation capability and IL-6 production, whereas the production of IL-4 and IL-10 is suppressed after co-culturing with MTECl. By contrast, MTECl could not induce thymocytes to secrete Thl type of cytokines. The results suggest that MTECl can regulate functional status of this thymocyte subset and induce them to develop into a specialized Th2 subset.     

热敏灸对人体免疫失衡影响的实验研究

募集17名受试者,随机分为对照组和热敏灸组,进行一周的高强度集训.期间,对照组每天运动后进行20 min的推拿按摩,热敏灸组每天运动后在双侧足三里处实施20 min的热敏灸治疗.采用ELISA法测定两组受试者集训前、末次训练后2 h与末次训练后3 d肘正中静脉血IFN-γ、IL-4、IL-12、IL-10、IL-12p40的含量;在热敏灸足三里治疗过程中,采集受试者红外热成像图形,分析热敏灸对穴位周围温度的影响.结果发现,热敏灸疗法可通过温热刺激和红外辐射等方式刺激穴位,提高IFN-γ/IL-4比值,调节IL-12及其抑制剂IL-10与IL-12p40的分泌,促进Th1细胞的分化过程,从而纠正过度疲劳所致的Th1/Th2失衡状态.     

Functional maturation of two murine medullary-type CD8SP thymocytes

TCRαβCD4-CD8+ thymocytes are heterogeneity. They may undergo phenotypic and functional maturation within thymic medulla. Medullary-type CD8SP thymocytes were divided into seven subsets based on phenotypic analysis, and their precursor-progeny relationship along with the differential pathway was also delineated. To further testify the validity of the maturation pathway, we purified 6C10-CD69+ cells representing the early stage and 6C10-Qa-2+ cells representing the later stage among medullary-type CD8SP thymocytes and compared their functional maturation levels. CD8+ T cells of spleen were used as the control. It is shown that there is no obvious difference of proliferation ability among these three subsets; however, intracytoplasmic cytokine assay shows that there is a hier archy of IFN-γ and TNFα secretion among these subsets, strikingly comparable to their phenotypic status among medullary type CD8SP thymocytes. The bioassays of IL-2 and IFN-γ in culture supernatant give the similar results.     

Quantitative detection of ING1 mRNA under different gene regulation based on molecular beacon

As an important tumor suppressor, normal expres- sion of ING1 is associated with the maintenance of cell normal physiological functions. Its low expression of- ten causes aberrant events of cell survive, growth and differentiation[1] which results in loss…     

Cytoskeleton reorganization and FAK phosphorylation are involved in lanthanum （Ⅲ）-promoted proliferation and differentiation in rat osteoblasts

The effect of lanthanum ion (La³⁺) on osteoblast function and cytoskeleton is assessed in vitro. Osteoblasts were isolated from Sprague-Dawley fetal neonatal rats. Cell proliferation and gene expression levels of cbfa-1, alkaline phosphatase (ALP), osteocalcin (OC), bone sialoprotein (BSP) and osteopontin (OPN) were examined by cell counting and RT-PCR. Cytoskeleton F-actin was stained with rhodamine-conjugated phalloidin and was visualized by a confocal microscope. As the results, 10⁻⁸-10⁻⁴ M La³⁺-induced osteoblast proliferation on day 2. Data from the RT-PCR assay revealed that 10⁻⁶ M La³⁺ up-regulated the expression levels of ALP, BSP, and cbfa-1 on day 4, while it enhanced the expressions of OC and OPN on day 21. The F-actin cytoskeleton was strengthened and reorganized under the exposure of La³⁺. In addition, the phosphorylation of focal adhesion kinase (FAK) was significantly promoted in 24 h evaluated by Western blot analysis. These findings indicate that La³⁺ promotes osteoblast activity through the phosphorylation of FAK and reorganization of the cytoskeleton.     

Signal transduction pathways mediated by colony stimulating factor-1 receptor

Colony-stimulating factor-1 (CSF-1), which is necessary for cell proliferation and differentiation, regulates both immediate and delayed early responses throughout G1 phase. The binding of CSF-1 to its receptor (CSF-1R) triggers phosphorylation of the receptor and its intrinsic tyrosine kinase. The activated receptor binds directly to cytoplasmic effector proteins, which induce multiple-signal transduction pathways. CSF-1 can induce the c-myc gene expression via Ras and Ets-related proteins. The expression of c-fos/jun family genes is also targeted following the activation of Ras. CSF-1R activates STAT1 and STAT3 to participate in signaling, but JAKs do not appear to contribute to signaling by CSF-1R. CSF-1R activates PI3-kinase, and PI3-ki can interact with downstream proteins by the MAPKK-related pathway independent of Ras/Raf. PC-PLC can enforce signaling in response to CSF-1. Furthermore, the turnover and dephosphorylation by the phosphatase SHPTP1 of CSF-1R are the major mechanism in the negative regulation of signaling by CSF-1R     

Construction and anti-tumor effects of recombinant fowlpox virus expressing Newcastle disease virus hemagglutinin-neuramidinase gene

Hemagglutinin-neuramidinase (HN), a Newcastle disease virus-derived protein, not only mediates receptor recognition but also possesses neuraminidase (NA) activity, the ability to cleave a component of those receptors, N-acetylneuraminic acid (NAcneu, sialic acid). It is known that this protein in mammalian species, including human beings, has interesting anti-neoplastic as well as immune stimulating properties. To explore the use of the HN gene in cancer gene therapy, we constructed a recombi-nant fowlpox virus expressing the HN protein (vFV-HN) and compared the anti-tumor activity of the recombinant virus with that of wild-type fowlpox virus (FPV) in vivo and in vitro. Here we found that although B16 cells were somewhat resistant to the basal cytotoxic effect of wild-type fowlpox virus, infection with vFV-HN caused a pronounced cytotoxic effect and, the survival of tumor-bearing mice immunized with vFV-HN was significantly increased compared with the survival of mice immunized with the FPV alone. Furthermore, the immunization of mice with vFV-HN elicited a B16 tumor-specific cytotoxic T lymphocyte (CTL) response and clonal expansion of both CD4 and CD8 T cell populations in vivo. In addition, T cells from lymph nodes of mice vaccinated with vFV-HN secreted high levels of the Th1 cytokine IL-2 and IFN-γ, indicating that the regression of tumor cells is related to a Th1-type dominant immune response. These results demonstrate that vaccination with vFV-HN may be a potential strategy for cancer gene therapy.     

Development of Th1-type immune responses requires the type I cytokine receptor TCCR

On antigen challenge, T-helper cells differentiate into two functionally distinct subsets, Th1 and Th2, characterized by the different effector cytokines that they secrete. Th1 cells produce interleukin (IL)-2, interferon-gamma (IFN-gamma) and lymphotoxin-beta, which mediate pro-inflammatory functions critical for the development of cell-mediated immune responses, whereas Th2 cells secrete cytokines such as IL-4, IL-5 and IL-10 that enhance humoral immunity. This process of T-helper cell differentiation is tightly regulated by cytokines. Here we report a new member of the type I cytokine receptor family, designated T-cell cytokine receptor (TCCR). When challenged in vivo with protein antigen, TCCR-deficient mice had impaired Th1 response as measured by IFN-gamma production. TCCR-deficient mice also had increased susceptibility to infection with an intracellular pathogen, Listeria monocytogenes. In addition, levels of antigen-specific immunoglobulin-gamma2a, which are dependent on Th1 cells, were markedly reduced in these mice. Our results demonstrate the existence of a new cytokine receptor involved in regulating the adaptive immune response and critical to the generation of a Th1 response.     

Immune responses of a designed HIV-1 DNA vaccine on rhesus monkeys

According to the UNAIDS/WHO report (http:// www.unaids.org/Epi2005/doc/report.html), there are about 40.3 million HIV/AIDS survivors, and the new HIV infection number amounts to about 4.9 million, while the death number is some 3.1 million in the world in…     

基于TCGA数据库分析UHRF1基因在恶性胸膜间皮瘤中的表达及预后意义

利用生物信息学数据库分析泛素样含PHD和环指结构域蛋白1（UHRF1）在恶性胸膜间皮瘤（MPM）中的表达水平及临床意义。基于TCGA数据库和GTEx数据库差异表达分析UHRF1 mRNA在MPM组织和正常肺组织中的表达水平;使用R软件分析UHRF1 mRNA表达量与临床病理参数的相关性;构建Kaplan-Meier模型和单因素多因素COX回归模型分析UHRF1基因在MPM中的预后;利用TIMER2.0数据库分析UHRF1基因与免疫细胞浸润的关系; GSEA分析UHRF1基因发挥功能的主要富集通路。选取8例MPM组织及4例非MPM胸膜组织,通过RT-qPCR的方法验证UHRF1在MPM与非MPM胸膜组织的表达情况。数据库分析结果表明,与正常肺组织相比,UHRF1 mRNA在MPM组织中高表达; UHRF1高表达患者提示MPM患者预后不良;UHRF1基因表达量与CD4⁺辅助型T细胞2、CD4⁺效应记忆性T细胞、巨噬细胞等多种免疫细胞浸润水平具有显著的相关性（P ＜ 0.01）,且显著影响MPM患者的预后。功能富集分析显示,UHRF1主要在DNA复制、蛋白酶体、同源重组等通路中起作用。在收集到的病例样本中,与非MPM胸膜组织相比,UHRF1 mRNA在MPM组织中的表达显著增高（P ＜ 0.001）。UHRF1在MPM组织中高表达,可能通过调节DNA甲基化和免疫细胞浸润来影响MPM患者预后,有望成为MPM治疗和预后评估的潜在靶点。     

Abrupt climate change of EastAsian Monsoon at 130 kaBP inferred from a high resolu-tion stalagmite δ18O record

^230Th ages and oxygen isotope data of a stalagmite from Shanbao Cave in Hubei Province characterize the East Asian Monsoon precipitation from 133 to 127ka. The decadal-scale high-resolution δ^18O record reveals a detailed transitional process from the Penultimate Glaciation to the Last Interglaciation. As established with ^230Th dates, the age of the Termination Ⅱ is determined to be 129.5±1.0 kaBP, which supports the Northern Hemisphere insolation as the triggers for the ice-age cycles. In our δ^18O record, the glacial/interglacial fluctuation reaches about 4‰, almost the same level as in other Asian Monsoon cave stalagmite δ^18O records. The transition of the glacial/interglacial period in our record can be recognized as four stepwise stages, among which, a rapid rise of monsoon precipitation follows the stage of “Termination Ⅱ pause”. The rapid rise is synchronous with the abrupt change of global methane concentration, which reflects that an increase in both Asian Monsoon precipitation and tropical wetland plays an important role in the global climate changes.     

Crystal structure and photoelectricity property of Fe(II/III) coordination supermolecules

Three new mixed-ligand Fe(II/III) complexes [Fe₂(μ₂–btec)(μ₂–H2btec)(phen)₂(H₂O)₂]_n (1), [Fe2(btec) (phen)₂(H₂O)₄] (2), and {[Fe(o-pha)(phen)(H₂O)]•H₂O}_n (3) (phen=1,10-phenanthroline, o-H₂pha=o-phthalic acid, H₄btec=1,2,4,5-benzenetetracarboxylic acid) have been hydrothermally synthesized and detected by single crystal X-ray diffraction, showing that complexes (1) and (2) are both bridged by the betc⁴⁻ ligands to form 1D chain and dinuclear structure and complex (3) is bridged by the o-pha groups to form 1D chain structure. The coordinated modes of the carboxyl groups adopt μ₂-η¹η¹ηη¹η¹ and μ₂-η²η¹ respectively in com-plexes (2) and (3). The betc groups in complex (1) show two different coordinated modes: μ₂-η¹η¹η¹η¹ and μ₂-η¹η¹. In addition, the hydrogen bonds and π…π type interactions make the complex molecule further connect to three-dimensional and two-dimensional networks respectively. These complexes are detected by IR, UV-Vis-NIR and surface photovoltage spectrum (SPS). The SPS of complexes (1)–(3) indicate that there are positive SPV responses in the range of 300–600 nm and show p-type semiconductor characteristic. Because the structure, the valence and the coordinative environment of the Fe ions are all different in the three complexes, the intensity, position and the number of the response bands are different obviously. The results of SPS are corresponding with UV-Vis-NIR spectra.