Role of macrophage colony-stimulating factor in the differentiation and expansion of monocytes and dendritic cells from CD34+ progenitor cells

The present study focused on whether it is possible to expand monocytic cells from CD34⁺ progenitor cells by using macrophage colony-stimulating factor (M-CSF) in the absence and presence of mast cell growth factor (MGF) and IL-6. It was demonstrated that CD34⁺ cells differentiate without expansion to functional mature monocytic cells in the presence of M-CSF or combinations of M-CSF plus IL-6 and MGF. A different response pattern was observed for the number of clonogenic cells. The addition of IL-6 or both IL-6 and MGF to M-CSF containing cultures resulted in significant higher numbers of colony-forming unit-macrophage (CFU-M) as tested in clonogenic and³H-thymidine assays. Furthermore, M-CSF plus both IL-6 and MGF appeared to be the most potent combination to preserve the monocytic precursor in cell suspension culture assays. These results indicate that IL-6 and MGF in conjunction with M-CSF affect CD34⁺ cells especially at precursor level without distinct effect on the more mature stages. Secondly we studied whether M-CSF is only critical for the monocytic lineage or also affects dendritic cell (DC) development. Indeed, we were able to culture CD83⁺ DC from CD34⁺ progenitor cells in the presence of M-CSF in conjunction with TNF-α, IL-4, and MGF although their absolute number is almost threefold lower than the number of CD83⁺ cells yielded from GM-CSF plus TNF-α, IL-4, and MGF stimulated CD34⁺ cells.     

A study of the suitability of gas chromatography-electron capture detection for the analysis of deoxynivalenol in cereals

A gas chromatographic-electron capture detection (GC-ECD) method for the analysis of deoxynivalenol (DON) in cereals was investigated. The sample was extracted with a mixture of acetonitrile-water and purified with a MycoSep #225 column. The silylation was performed with Tri-Sil-TBT reagent, followed by dilution with hexane and a washing step with buffer. By using Tri-Sil-TBT reagent no double peaks were observed for DON in the gas chromatograms, in comparison with two other silylation reagents TMSI and Tri-Sil-Z. The use of trichothecolone (TRI) as an internal standard for DON was studied in order to indicate possible problems in the derivatisation reaction. TRI proved to be a relatively good internal standard for DON in cereal samples, as well as 1,1-bis-(4-chlorophenyl)-2,2-dichloroethylene (DDE), which was used as a GC standard for ensuring the function of GC-ECD. During the study, a matrix effect was clearly observed between the cereal matrix-assisted calibration curve and the calibration curve prepared without cereal matrix. The results of spiked and reference material samples, quantified with the calibration curve prepared without and with matrix, demonstrated that the matrix affects the results. However, after recovery correction the results were comparable. The validation results demonstrated that the GC-ECD method for DON analysis in cereals is sufficiently reliable.     

CpG联合铝佐剂对HCV免疫原体液免疫作用的影响

目的:探讨胞苷酸鸟苷寡脱氧核苷酸(CpG ODN)联合铝佐剂对丙型肝炎病毒(HCV)重组免疫原的体液免疫作用。方法:采用高交叉HCV-HVR1和E1重组蛋白与CpG ODN、铝佐剂组合,免疫BALB/c小鼠后以ELISA、酶联免疫斑点测定、流式细胞术、免疫沉淀等方法检测相关体液免疫指标和免疫血清多抗的交叉反应性。结果:CpG联合铝佐剂激发了最高的特异性抗体滴度;佐剂通过提高抗体分泌细胞数量、增加脾脏中记忆B细胞数量、增加脾淋巴细胞IL-6、IL-10分泌浓度实现体液免疫增效;CpG则能提高免疫效率,联合铝佐剂时显著提高浆细胞数量;12份HCV阳性血清中有10份可与多抗HVR1 IgG发生免疫沉淀。结论:CpG和铝佐剂联合应用具有协同作用,多抗HVR1 IgG具有较好的交叉反应性。     

HLA-DRB1 shared epitope genotyping using the revised classification and its association with circulating autoantibodies,acute phase reactants,cytokines and clinical indices of disease activity in a cohort of South African rheumatoid arthritis patients

Introduction  

The revised shared epitope (SE) concept in rheumatoid arthritis (RA) is based on the presence (S) or absence (X) of the SE RAA amino acid motif at positions 72 to 74 of the third hypervariable region of the various human leucocyte antigen (HLA)-DRB1 alleles. The purpose of this study was to investigate SE subtypes on the basis of the American College of Rheumatology 1987 revised criteria for the classification of RA in a cohort of South African RA patients (n = 143) and their association with clinical and circulating biomarkers of disease activity (autoantibodies, acute phase reactants and cytokines).     

一种新的制备多克隆抗体的方法

用亲和色谱纯化的rhEPO作为抗原免疫KM小鼠和Bal b/c小鼠,然后腹腔注射S180细胞或SP2/0细胞。S180细胞可刺激KM小鼠产生腹水,腹水量大,抗体效价高。而直接注射SP2/0细胞既未能使KM小鼠产生腹水,也未能使Bal b/c小鼠产生腹水。实验提供了一种新的简便、快速、经济地制备高效价多克隆抗体的方法。     

High risk populations and HIV-1 infection in China

INTRODUCTION HIV has spread to all of China’s 31 provinces, autono- mous regions and municipalities, creating one of the fast- est-growing HIV/AIDS epidemics in the world [1,2]. The HIV/AIDS epidemic in China has gone through three phases: the Entry Phase (1985 -1988), the Spreading Phase (1989-1994) and the Expansion Phase (1995- present). The striking increase of HIV-1 infections over the past few years may herald entry into a new fourth phase that will include much larger nu…     

Antiplatelet and anticoagulant therapy in elective percutaneous coronary intervention

Thrombosis plays a major role in acute vessel closure both after coronary balloon angioplasty and after stenting. This review will address the role of antiplatelet and anticoagulant therapy in preventing early thrombotic complications after percutaneous coronary intervention. The focus will be on agents that are routinely available and commonly used.     

The Chlamydia muridarum-induced IFN-β response is TLR3-dependent in murine oviduct epithelial cells

Epithelial cells lining the murine genital tract act as sentinels for microbial infection, play a major role in the initiation of the early inflammatory response, and can secrete factors that modulate the adaptive immune response when infected with Chlamydia. C. muridarum-infected murine oviduct epithelial cells secrete the inflammatory cytokines IL-6 and GM-CSF in a TLR2-dependent manner. Further, C. muridarum infection induces IFN-β synthesis in the oviduct epithelial cells in a TRIF-dependent manner. Because murine oviduct epithelial cells express TLR3 but not TLRs 4, 7, 8, or 9, we hypothesized that TLR3 or an unknown TRIF-dependent pattern recognition receptor was the critical receptor for IFN-β production. To investigate the role of TLR3 in the Chlamydia-induced IFN-β response in oviduct epithelial cells, we used small interfering RNA, dominant-negative TLR3 mutants, and TLR3-deficient oviduct epithelial cells to show that the IFN-β secreted during C. muridarum infection requires a functional TLR3. Interestingly, we demonstrate that the TLR3 signaling pathway is not required for IFN-β synthesis in C. muridarum-infected macrophages, suggesting that there are alternate and redundant pathways to Chlamydia-induced IFN-β synthesis that seem to be dependent upon the cell type infected. Finally, because there is no obvious dsRNA molecule associated with Chlamydia infection, the requirement for TLR3 in Chlamydia-induced IFN-β synthesis in infected oviduct epithelial cells implicates a novel ligand that binds to and signals through TLR3.     

人呼吸道合胞病毒兰州株基因组全长cDNA克隆的构建及鉴定

目的构建用于呼吸道合胞病毒(respiratory syncytial virus,RSV)体外拯救的RSV基因组全长cDNA克隆,并进行鉴定。方法根据RSV Long株基因组序列设计并合成引物,利用RT-PCR技术分6段扩增RSV LZ01/09基因组序列并构建克隆载体;测序后,利用重叠PCR与酶切连接技术,根据基因组序列选择特异性酶切位点,引入Kpn I、Xma I和Sal I酶切位点,构建成4个亚克隆载体;将亚克隆载体的插入片段连接至经过改造且包含T7启动子、锤头状核酶、多克隆位点、丁肝核酶、T7终止子的p RSV1载体中,构建RSV基因组全长cDNA克隆;对克隆全长cDNA序列进行测定,与亲本RSV LZ01/09基因组进行同源性比对分析,并与RSV实验参比株进行系统进化树分析。结果测序结果显示,RSV LZ 01/09的基因组全长为15 204 bp,与GenBank公布的RSV基因组序列长度相当,将完整的序列提交GenBank,登录号为KY782635;酶切及测序结果显示,用于RSV全长cDNA克隆构建的基本载体p BSKS-MCS(简称p RSV1)与预期相符,RSV全长基因组cDNA克隆质粒(简称转录载体p RSV1-4F)酶切片段大小与预期一致;同源性比对结果显示,全长cDNA序列与亲本RSV LZ01/09基因组序列同源性高达99.83%;系统进化树分析结果显示,其与RSV-A亚型序列同属于一个分支。结论测序及酶切分析结果表明已成功构建RSVLZ01/09基因组全长cDNA克隆,为建立拯救RSV重组病毒的反向遗传学系统平台奠定了基础。