Histamine downregulates CD14 expression via H2 receptors on human monocytes

Lipopolysaccharide (LPS) binds to LPS-binding protein (LBP) in plasma and is delivered to the cell surface receptor CD14 on human monocyte. LPS is transferred to the transmembrane signaling receptor toll-like receptor (TLR) 4. In the present study, the effect of histamine on the expression of CD14 on human monocytes was investigated. Histamine concentration- and time-dependently decreased the expression of cell surface CD14, whereas histamine did not decrease mRNA for CD14 nor increase soluble CD14 (sCD14). The inhibitory effects of histamine on CD14 expression were antagonized by H2-receptor antagonist, but not by H1 and H3/H4 antagonist. The effects of selective H2-receptor agonists, 4-methylhistamine and dimaprit, on CD14 expression mimicked that of histamine indicating that histamine regulated CD14 expression through the stimulation of H2-receptors. The pretreatment with histamine partially inhibited the LPS-induced TNF-alpha production in human peripheral blood mononuclear cells (PBMC). Such inhibition might be due to the down-regulation of CD14 expression on monocytes by histamine.     

Toll-like receptor 9 contributes to recognition of Mycobacterium bovis Bacillus Calmette-Guérin by Flt3-ligand generated dendritic cells

Recognition of mycobacteria by the innate immune system is essential for the development of an adaptive immune response. Mycobacterial antigens stimulate antigen presenting cells (APCs) through distinct Toll-like receptors (TLRs) resulting in rapid activation of the innate immune system. The role of TLRs during infection with Mycobacterium bovis Bacillus Calmette-Guérin (BCG) has been evaluated for TLR2 and TLR4 only. Surprisingly, despite the fact that immune stimulatory CpG-motifs have been originally derived from BCG, for the vaccine strain the role of TLR9 has not been addressed before. To identify the set of TLRs involved in the recognition of BCG, we infected bone marrow-derived macrophages and bone marrow-derived dendritic cells (Flt3-ligand generated DCs) from TLR2, TLR3, TLR4, TLR7, TLR9, MyD88 knockout, TLR2/4 and TLR2/4/9 multiple knockout mice. The degree of activation and stimulation was determined by TNFα, IL-6 and IL-12p40 ELISA. Activation of DCs was measured by surface expression of the costimulatory molecule CD86. We observed the most dramatic reduction of the inflammatory response for TLR2-deficient antigen presenting cells. Both macrophages and DCs produce markedly decreased amounts of TNFα and IL-6 in the absence of TLR2 whereas no significant reduction could be observed for TLR3, 4, 7, 9 single TLR-knockouts. However, IL-12 production in DCs appears not exclusively dependent on TLR2 and only in TLR2/4/9-deficient DCs BCG-induced IL-12 is reduced to background levels. Similarly, up-regulation of CD86 is abolished only in TLR2/4/9-deficient DCs supporting a role of TLR9 in the recognition of M. bovis BCG by murine dendritic cells.     

IL-4对DC产生IL-12影响的研究

为了研究白细胞介素 4 (IL 4 )对树突状细胞 (dendriticcell,DC )产生白细胞介素 1 2 (IL 1 2 )p70的调节作用及其机制 ,将小鼠骨髓细胞在含GM CSF和IL 4的培养液中培养 6d ,加入成熟诱导剂脂多糖 (LPS )或TNF α、CpG ODN继续培养 2d以获得成熟DC。流式细胞仪检测DC表面CD80和MHCII类分子 ,混合淋巴细胞反应检测DC促进同种异体T细胞增殖的能力 ,ELISA法检测不同诱导剂诱导DC产生IL 1 2p70的能力 ,RT PCR和荧光定量PCR检测IL 1 2p35和p4 0mRNA的表达及其变化。结果显示小鼠骨髓细胞在含GM CSF、IL 4和成熟诱导剂的培养液中培养后可以获得成熟的DC ,成熟DC高表达CD80和MHCII类分子 ,具有较强的刺激同种异体T细胞增殖的能力 ,LPS、CpG ODN、TNF α、PolyI:C在促进DC成熟的同时能诱导DC产生有活性的IL 1 2 ,IL 4对IL 1 2的产生具有明显的促进作用 ,RT PCR和荧光定量PCR结果显示LPS诱导IL 1 2产生以及IL 4对其的促进作用与IL 1 2p35基因的转录水平增高有关     

Detection of bloodstream pathogens in a bacille Calmette-Guérin (BCG)-vaccinated pediatric population in Malawi: a pilot study

Children in Malawi receive bacille Calmette-Guérin (BCG) vaccination within the first 3 days of life. Thus, we hypothesized that Malawian children infected with the human immunodeficiency type 1 virus (HIV-1) might be particularly vulnerable to dissemination of the BCG Mycobacterium bovis strain with which they were vaccinated. Following informed consent by parents, we studied children admitted to a Malawi general hospital during the 1998 wet and dry seasons. Blood from cohorts of acutely ill children was cultured for bacteria, including mycobacteria, and fungi, and tested for anti-HIV-1 antibodies. It was shown that non- typhi Salmonella and Escherichia coli were the predominant bloodstream pathogens during the wet and dry seasons, and that bloodstream dissemination of the BCG M. bovis strain is uncommon in HIV-1-infected children who receive the BCG vaccine.     

Recombinant Mycobacterium bovis bacillus Calmette-Guérin (BCG) expressing mouse IL-18 augments Th1 immunity and macrophage cytotoxicity

Interleukin-18 (IL-18) has been demonstrated to synergize with BCG for induction of a T-helper-type 1 (Th1) immune response. Since successful treatment of superficial bladder cancer with BCG requires proper induction of Th1 immunity, we have developed a recombinant (r) BCG strain that functionally secretes mouse (m) IL-18. This rBCG-mIL-18 strain significantly increased production of the major Th1 cytokine IFN-gamma in splenocyte cultures, at levels comparable to that elicited by control BCG plus exogenous rIL-18. IFN-gamma production by splenocytes was eliminated by addition of neutralizing anti-IL-18 antibody. Endogenous IL-12 played a favourable role whereas IL-10 played an adverse role in rBCG-mIL-18-induced IFN-gamma production. Enhanced host antimycobacterial immunity was observed in mice infected with rBCG-mIL-18 which showed less splenic enlargement and reduced bacterial load compared to control mice infected with BCG. Further, splenocytes from rBCG-mIL-18-infected mice, in response to BCG antigen, displayed increased production of IFN-gamma and GMCSF, decreased production of IL-10, elevated cellular proliferation and higher differentiation of IFN-gamma-secreting cells. rBCG-mIL-18 also enhanced BCG-induced macrophage cytotoxicity against bladder cancer MBT-2 cells in a dose-dependent manner. Neutralizing all endogenous macrophage-derived cytokines tested (IL-12, IL-18 and TNF-alpha) as well as IFN-gamma severely diminished the rBCG-mIL-18-induced macrophage cytolytic activity, indicating a critical role for these cytokines in this process. Cytokine analysis for supernatants of macrophage-BCG mixture cultures manifested higher levels of IFN-gamma and TNF-alpha in rBCG-mIL-18 cultures than in control BCG cultures. Taken together, this rBCG-mIL-18 strain augments BCG's immunostimulatory property and may serve as a better agent for bladder cancer immunotherapy and antimycobacterial immunization.     

公立医院优势学科病种绩效管理研究:以骨科为例

目的从循证视角建立公立医院优势学科病种绩效管理工具。方法以波士顿矩阵理论为指导,以“病种均次毛利率、病种均次住院日”为主要指标,“日均毛利率”为辅助指标,采用Microsoft office EXCEL 365 软件进行分析。结果2019年骨科平均住院日为4.99 d,均次平均毛利率为23.62%,日均毛利率平均值为4.73%。骨科200例以上病种、不同亚专科病区病种、同一亚专科病区内不同病种、不同亚专科病区相同病种,绩效均存在差异。结论 管理者应对不同病种绩效指标进行适当校正,探索精细化病种诊疗与管理模式。     

BT—BAK细胞和BCG膀胱腔内过继免疫治疗对膀胱癌患者全身免疫功能影响的比较

目的：观察BT-BAK细胞与BCG膀胱腔内灌注治疗对患者全身免疫功能的影响。方法：由手术切除的膀胱瘤标本制备膀胱肿瘤可溶性抗原,以BCG为佐剂,从PBMNC中诱导出抗膀胱肿瘤特异性细胞毒性BT-BAK细胞,临床选择72例浅表性膀胱癌术后患者随机平均分成2组分别进行BT-BAK和BCG的膀胱腔内灌注治疗。于不同阶段采用ELISA法测定患者外周血中IL-2、TNF-α及IFN-γ的含量;观察治疗后患者PBMNC对T24膀胱肿瘤细胞株杀伤活性的变化。结果：BT-BAK组患者血清中IL-2、TNF-α及IFN-γ的水平均较治疗前明显提高（P＜0.01);患者PBMNC对T24膀胱肿瘤细胞株的杀伤活性显著增强（P＜0.01)。BT-BAK组上述指标均高于BCG组（P＜0.05)。平均随访18.16个月,两组的复发率分别为2.7%和11.11%。结论：BT-BAK细胞及其培养上清进行膀胱腔内灌注治疗,能够有效提高膀胱癌术后患者的全身免疫水平,降低膀胱癌的术后复发率。     

泰宁县584名儿童结核菌素试验结果分析

目的：了解我县儿童接种卡介苗接种的免疫情况。方法：1999年7－8月,以城区范围内卡介苗接种16周以后的1－4岁儿童为对象进行结核菌素试验,72小时后查验反应结果。注射部位硬结反应≥5mm为阳性反应,同时记录儿童接种卡俚苗后的卡痕大小。结果：我县城区1－4儿童结核菌素试验阳转率为45．9％,其中0－岁组儿童结核菌素是性转变达88．9％,对结核病的免疫水平较低。结核菌素试验阳性率性别间无显性差异。止痕＜5mm的儿童,1－4岁年龄组的结核菌素阳性转率逐渐增大,结论：城区中存在一定的传染源,对阳性转率较低的原因应进行探索并提出预防方法和措施。