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.     

北京农村居民健康状况及卫生服务需求调查

目的了解北京农村居民健康状况及卫生服务需求特点。方法从昌平区、顺义区、大兴区、房山区随机抽取25～64岁农民1605人进行统一问卷调查。结果农民高血压自报患病率为22．4％，超重和肥胖率为56．1％；现在吸烟率为34．4％，缺乏运动率为41．6％；家庭主要经济支出为学生上学(35．3％)，建房(25．4％)和疾病(21．2％)；54．9％家庭医疗费用支出占总收入的10％以上；最关心的健康问题主要是慢性疾病防治(70．2％)；最希望获得的健康知识是慢性病预防知识(72％)；希望获得健康知识的途径是广播和电视(81％)；家庭最希望获得的医疗保健服务是方便看病和获得药物(73．8％)。结论慢性病给农民带来沉重负担，并成为农民关心的主要健康问题。     

腺苷对突触体^45Ca摄取的抑制作用

用同位素标记~(15)Ca法测定大鼠突触体内游离钙的浓度,探讨了腺苷对高钾、N-甲基-D-天冬氨酸(NMDA)谷氨酸(Glu)等刺激所致的Ca~(2 )内流增加的影响。结果发现腺苷在10nmol/L～0.1μmol/L范围内对高钾、NMDA、Glu等刺激所致的Ca~(2 )内流有抑制作用,并呈剂量依赖关系,最大抑制率分别为41.68±7.68％、31.32±6.17％、37.52±2.29％。这可能是腺苷对缺血性脑损伤保护作用的机理。     

妊娠期肝内胆汁淤积症与血清甘胆酸的关系

检测了７４例正常足月妊娠妇女和５６例妊娠期肝内胆汁淤积症（ＩＣＰ）患血清甘胆酸（ＣＧ）的浓度，结果ＩＣＰ患血ＣＧ浓度显升高（１３．５２±９．５４ｍｇ／Ｌ），为正常孕妇（２．６７±１．９２ｍｇ／Ｌ）的５～１０倍，根据血ＣＧ浓度将ＩＣＰ患分为〈１０、１０～２０，〉２０ｍｇ／Ｌ３组，将３组胎儿出生情况与正常组相对照，结果显示血ＣＧ浓度〈１０ｍｇ／Ｌ组羊水胎粪污染率及新生儿窒息率与正常组无差异，而     

大鼠胎胰胰岛组织培养及移植的实验观察

本实验对培养中的大鼠胎胰胰岛组织及其移植后的形态功能进行了观察。结果表明,大鼠胎胰经短期培养,不仅内分泌胰岛β细胞的活力保存良好,而且胰外分泌腺泡大大减少以致消失。21只糖尿病大鼠,同种移植经过7天培养的胎胰胰岛,未用免疫抑制剂,术后糖尿病缓解率76.2％。     

脑血管血液动力学参数脑卒中预测模型的建立

目的 根据脑血管血液动力学参数(CVHI)和脑卒中的主要危险因素建立脑卒中预测模型。方法 选择全国六大行政区脑卒中研究队列人群25355例，将基线调查时的CVHI检测结果 进行主成分分析，再以各主成分和主要脑卒中危险因素为自变量，以随访中脑卒中发病为应变量进行回归分析，根据回归系数建立脑卒中预测模型，计算发病概率，绘制ROC曲线，确定最佳截断点，评价预测模型的预测效能。结果 四个主成分的累积贡献率依次为58．1％、79．4％、88．4％和94．6％，被筛检进入logistic回归方程的变量分别为第一至第四主成分、高血压病史、年龄和性别，ROC曲线下面积为0．855，最佳截断点为预测概率≥0．05，预测脑卒中的敏感度、特异度和准确度分别为踟．7％，78．5％，78．5％。结论 通过主成分回归分析，可以建立具有良好效能的脑卒中预测模型。     

Clinical pathologic study on effect of Qianggan capsule in treating patients of chronic hepatitis B with liver cirrhosis

Objective  

To explore the clinical therapeutic effect of Qianggan Capsule (QGC) in treating chronic hepatitis B with liver fibrosis from the pathological aspect.     

直接冠状动脉内支架安置术和r-SK静脉溶栓治疗急性心肌梗死的对比研究

①目的　比较急性心肌梗死 (AMI)病人直接冠状动脉内支架安置和重组链激酶 (r SK)静脉溶栓治疗的临床疗效。②方法　对初次发病 1 2h以内的 1 0 5例AMI病人 ,随机给予直接冠状动脉内支架安置 (支架组 ,6 0例 )和静脉r SK溶栓 (溶栓组 ,5 5例 )治疗。观察两组梗死相关血管再通成功率、心肌梗死溶栓试验 (TIMI)Ⅲ级血流发生率、平均住院天数、住院病死率 ,出院前二维超声心动图测定并推算左心室射血分数 (LVEF)和梗死区室壁运动指数 (RWMI) ,出院后 6个月病死率、LVEF和RWMI。③结果　支架组与溶栓组相比较 ,梗死相关血管再通率显著提高、TIMIⅢ级血流发生率显著增加、平均住院天数显著缩短、住院病死率显著降低 (χ2 =5 .4 2 4～2 8.931 ,t=7.90 1 ,P <0 .0 5、0 .0 1 ) ;出院前LVEF和RWMI ,两组间存在显著差异 (t =3.2 91、2 .1 2 7,P <0 .0 5、0 .0 1 ) ;出院后 6个月病死率、LVEF和RWMI ,两组比较无显著差异。出院后 6个月LVEF、RWMI与出院前比较 ,支架组与溶栓组均有显著改善 (t=2 .1 92～ 4 .6 1 1 ,P <0 .0 5、0 .0 1 )。④结论　与静脉r SK溶栓治疗相比 ,直接冠状动脉内支架安置术治疗AMI,可更有效地再通梗死相关血管、降低住院病死率、更有效地保护病人心脏功能     

经直肠超声引导前列腺穿刺活检203例临床分析

目的评估经直肠超声引导的前列腺六针穿刺活检在前列腺癌及前列腺其他疾病的诊断和鉴别诊断的价值。方法对指肛检查阳性，血清PSA〉4pg／L及经直肠超声检查前列腺声像图异常怀疑有占位性病变的203人进行经直肠超声引导的前列腺穿刺活检。结果穿刺活检的203例病理结果：良性前列腺增生（BPH）104例占51．24％，前列腺癌（PCa）95例占46．80％，前列腺结核及前列腺平滑肌肉瘤各2例，分别占0．98％。结论经直肠超声引导的前列腺穿刺活检其操作简单，病人痛苦小，并发症少，较安全。在前列腺癌及其他前列腺疾病的诊断与鉴别诊断中有重要的临床价值。     

抗端粒酶核酶抑制肝癌细胞裸鼠移植瘤生长的研究

设计针对端粒酶RNA组分的特异性核酶，研究该核酶对肝癌细胞裸鼠移植瘤生长的影响。构建含针对端粒酶RNA组分锤头状核酶基因的重组真核表达质粒pBBS212-RZ，以及建立人肝癌细胞株HepG2裸鼠移植瘤模型。将不同剂量的pBBS212-RZ用脂质体包裹后进行瘤体内多点注射，对照组注射生理盐水或空质粒载体pBBS212。连续注射2l天后，测量移植瘤体积和重量，检测瘤组织端粒酶活性。抗端粒酶特异性核酶使瘤组织端粒酶活性下降(抑制率为72％)，明显地抑制了肝癌细胞裸鼠移植瘤生长(抑制率为68％)，且存在剂量依赖性。抗端粒酶特异性核酶作为一种有效的端粒酶抑制剂，可抑制肝癌细胞的生长，有望在肝癌基因治疗中发挥作用。