Lethal effect of mononuclear cells derived from human umbilical cord blood differentiating into dendritic cells after in vitro induction of cytokines on neuroblastoma cells

BACKGROUND: Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human granulocyte-macrophage colony stimulating factor (rhG-MCSF) and recombinant human interleukin-4(rhIL-4) can generate a great many dendritic cells and promote the lethal effect of T cells on human neuroblastoma, but it is unclear that whether the lethal effect is associated with the most proper concentration of dendritic cells. OBJECTIVE: To investigate the lethal effect of human umbilical cord blood mononuclear cells induced in vitro by cytokines differentiating into dendritic cells on human neuroblastoma, and its best concentration range. DESIGN: Open experiment. SETTING: Department of Pediatrics, the Medical School Hospital of Qingdao University. MATERIALS: The study was carried out in the Shandong Provincial Key Laboratory (Laboratory for the Department of Pediatrics of the Medical School Hospital of Qingdao University) during September 2005 to May 2006. Human umbilical cord blood samples were taken from the healthy newborn infants of full-term normal delivery during October to November 2005 in the Medical School Hospital of Qingdao University, and were voluntarily donated by the puerperas. Main instruments: type 3111 CO2 incubator (Forma Scientific, USA), type 550 ELISA Reader (Bio-Rad, USA). Main reagents: neuroblastoma cell line SK-N-SH (Shanghai Institute of Life Science, Chinese Academy of Sciences), RPMI-1640 culture fluid and fetal bovine serum (Hyclone), rhIL-4 (Promega, USA), rhG-MCSF (Harbin Pharmaceutic Group Bioengineering Co.Ltd), rat anti-human CD1a monoclonal antibody and FITC-labeled rabbit anti-rat IgG (Xiehe Stem cell Gene Engineering Co.Ltd). METHODS: ① Human umbilical cord blood mononuclear cells obtained with attachment methods differentiated into human umbilical cord blood dendritic cells, presenting typical morphology of dendritic cells after in vitro induction by rhG-MCSF and rhIL-4. ② Different concentrations of dendritic cells[ dendritic cells: neuroblastoma cells=20∶1,50∶1,100∶1（2×108 L-1，5×108 L-1，1×109 L-1）], 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the experimental group. 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the control group. ③ Main surface marker CD1a molecules of dendritic cells were detected with indirect immunofluorescence, and the percent rate of dendritic cells was counted with ultraviolet light and expressed as the expression rate of CD1a+ cells. ④ Single effector cells and target cells were respectively set in the experimental group and control group to obtain the lethal effect. The lethal effect of dendritic cells on neuroblastoma cells was indirectly evaluated by detecting cellular survival with MTT assay. The lethal effect(%)=（1-A experimental well-A effector cell well/A target cell well）×100%.⑤The experimental data were presented as Mean ±SD, and paired t test was used. MAIN OUTCOME MEASURES: ① Morphological characters of dendritic cells in the process of induction and differentiation. ②CD1a+ cellular expression rate. ③Lethal effect of dendritic cells on neuroblastoma cells. RESULTS: ①Morphological characters of dendritic cells in the process of induction and differentiation: On the 15th day after human umbilical cord blood mononuclear cells were induced by rhG-MCSF and rhIL-4, typical morphology of dendritic cells could be seen under an inverted microscope. ②Expression rate of CD1a+ cells was （43.12±5.83）%. ③Lethal effect of dendritic cells on neuroblastoma cells: Lethal effect of dendritic cells stimulated T cells in each experimental group ( dendritic cells: neuroblastoma cells=100∶1,50∶1,20∶1 respectively) on neuroblastoma cells was significantly higher than that in control group[（31.00 ±4.41）%，（30.92±5.27）%，(33.57±5.35)%,(26.23±5.20)%, t=3.51，2.98，4.24, P < 0.01）; But the lethal effect of dendritic cells on neuroblastoma was significantly lower when their ratio was 100∶1 and 50∶1 in comparison with 20:1 （t=2.01，2.36, P < 0.05）, and no significant difference in lethal effect existed between the ratio at 100∶1 and 50∶1（t=0.06,P > 0.05）. CONCLUSION: Dendritic cells differentiated from human umbilical cord blood mononuclear cells after in vitro induction of cytokines can promote the lethal effect of T cells on neuroblastoma cells. The lethal effect is associated with the concentration of dendritic cells within some range.     

扩散法被动式甲醛个体监测器（Ⅱ型）的研制

介绍了一种基于气体分子扩散原理的被动式甲醛个体采样器，它是在Ⅰ型采样器的基础上完成的。采用甘油（２０％）／偏重亚硫酸钠（０．２５％）浸渍的定量滤纸（φ４２ｍｍ）作为吸收层，空气中甲醛扩散到吸收层上，形成稳定的化合物。采样一定时间后，取出吸收层，洗脱后，用ＡＨＭＴ化学比色法测定所采集到的甲醛。这种采样器的平均采样速率为８３ｍｌ／ｍｉｎ，标准偏差为７．２ｍｌ／ｍｉｎ，与有动力吸收管采样的方法相比较，总不确定度小于±２５％。     

县级医院经济活力及其与不同奖金制度之间的关系分析

该利用山东省县级医院普查资料，通过比较性分析方法，对奖金制度转变和医院经济活力之间的关系进行了分析。研究结果表明：在过去的20年中，医院经济活力得到了较大提高，奖金制度转变有利于医院提高经济活力。     

致倦库蚊有机磷抗性相关扩增酯酶B基因的多样性

本实验从佛山和成都两地的致倦库蚊群体中筛选出３个酯酶类型不同的有机磷（ＯＰ）抗性品系，ＦＳ－１、ＦＳ－２和ＣＤ－１品系。各品系的酯酶基因Ｂ扩增水平、酯酶活性和ＯＰ抗性水平三者一致。各品系酯酶Ｂ基因限制性酶切片段比较分析表明，我国的ＯＰ抗生库蚊群体中不仅有世界性分布的酯酶Ｂ１和Ｂ２，而且在佛山和成都地区各有一个新的独立扩增的酯酶Ｂ，分别命名为Ｂ６和Ｂ７。     

亚胺培南/西司他丁与头孢他啶治疗中、重度细菌感染疗效费用分析比较研究

对我院４５例确诊为中、重度细菌感染住院患者进行了亚胺培南／西司他丁与头孢他啶疗效费用分析比较研究。结果表明：２组病例有效率、死亡率无显著性差异；头孢他啶组较亚胺培南／西司他丁组疗程明显延长。亚胺培南／西司他丁每日所需费用明显高于头孢他啶；治疗结束时，前者全部费用并未超过后者；全部住院费用无明显差异。作者认为：决定２种药物全部费用的因素，除与药物单价和每日费用有关，还与药物疗程密切相关。选用药物抗菌作用越强，用药时间即相应缩短，住院时间必然缩短；最终患者住院费用降低     

Transformation of immortal, non-tumorigenic osteoblast-like human osteosarcoma cells to the tumorigenic phenotype by nickel sulfate

Epidemiological studies have indirectly linked compounds ofchromium, nickel and arsenic to human carcinogenesis. However,there is no evidence that metal compounds can transform humancells to the tumorigenic phenotype in culture. We show herethat exposure to 36 µM NiS0₄ for 48–96 h resultsin transformation of an immortal, non-tumorigenic, osteoblast-likecell line, HOS TE85, to the tumorigenic phenotype. Continuouspassaging following treatment leads to the formation of a fewdense foci. The cells isolated and expanded from the foci aremorphologically transformed, and form anchorage-independentcolonies of the size and abundance comparable to that formedby Kirsten murine sarcoma virus transformed HOS TE85 cells.The transformed cells from tumors in nude mice, have enhancedlevels of plasminogen activators and have lost the ability toform model bone matrix on extended culture in the presence ofascorbic acid and ß-glycerophosphate. A number ofcell lines have been established from nude mouse tumors. Cytogeneticanalysis reveals 16 marker chromosomes and an aberrant chromosome16. This is the first report of the transformation of a humancell line to tumorigenic phenotype by a metal carcinogen.     

脑外科手术后发生正常灌注压突破的发病机制及超微病理基础

目的:探讨脑外科手术后部分患者病变邻近脑组织发生正常灌注压突破并发症的发病机制及其超微病理基础.材料和方法:对68例脑内血管畸形病变邻近的脑组织进行电子显微镜观察.结果:病灶邻近脑组织内部分毛细血管外周的星形细胞足突缺失或出现血管基膜疏松、分层等发育不良的情况,甚至有些血管壁组织结构破损.结论:电子显微镜检查证实,在血管畸形病灶周围的脑组织内可见病理性血管.该血管的基膜发生病变,血管周围的星形细胞足突明显减少或破坏.当血液灌注压力突然变化时,这种病变的血管就可能发生液体外渗及破裂.这就是在神经外科手术中或手术后发生正常灌注压突破并发症的超微病理基础.     

Toluene vapor exposure and urinary excretion of hippuric acid among workers in China.

A factory survey was conducted in three provinces in China from 1985 to 1989. The time-weighted average toluene concentrations in breathing zone air were monitored by diffusive sampling, whereas hippuric acid (HA) concentrations in shift-end urine samples were measured by high performance liquid chromatography (HPLC). Exposed workers (456 men and women) were those for whom toluene (up to 548 ppm toluene) accounted for greater than or equal to 90% of total exposure (by vapor concentration in ppm), whereas 517 nonexposed controls were recruited from the same factories or from factories of the same region. There was a linear correlation between the intensity of toluene exposure and HA concentration in the shift-end urine. Comparison of the results with findings in the literature shows that the toluene-induced increase in urinary HA concentration among workers in China is significantly smaller than the published values, whereas HA concentrations in urine samples from nonexposed controls are comparable to the levels previously reported.