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.     

cDNA probes for the diagnosis of bovine torovirus (Breda virus) infection.

A genomic cDNA library of RNA from Breda virus (BRV), a bovine torovirus, was prepared. The nucleotide sequence of the 3' end of the genome was found to be highly conserved (93% identical) between BRV and Berne virus, the torovirus prototype. Cross-hybridization experiments were performed to select Berne virus cDNA clones for use as probes in a dot hybridization assay; the objective was to detect heterologous torovirus RNA in fecal material. A rapid RNA extraction method was employed to make the test applicable for routine diagnosis. Samples from calves after experimental and natural infection with BRV were assayed to establish the sensitivity and specificity of the test and to compare the test with the enzyme-linked immunosorbent assay (ELISA) for antigen detection. For this purpose, 53 samples from seven infected calves were tested with both methods. In the ELISA, BRV was detected in six fecal samples from three inoculated calves. By use of the hybridization test, 16 samples from seven calves reacted positively. With one exception, only postinoculation samples were found positive in hybridization. No signal was seen in feces from uninoculated calves or from calves infected with rotavirus or coronavirus.     

Dynamics, structure, and function are coupled in the mitochondrial matrix.

The coupling between molecular diffusion and the structure and function of the rat liver mitochondrial matrix was explored using fluorescence anisotropy techniques and electron microscopy. The results confirm that matrix ultrastructure and the concentration of matrix protein are influenced by the respiratory state of mitochondria and the osmolarity of the external medium. At physiological osmolarity, a fluorescent metabolite-sized probe was found to diffuse slowly in the mitochondrial matrix but not to be completely immobile. In addition, significant differences in diffusion rates were found to exist between different mitochondrial respiratory states, with the slowest diffusion occurring in states with the highest matrix protein concentration. These data support the concept of a matrix structure in which diffusion is considerably hindered due to limited probe-accessible water and further suggest that volume-dependent regulation of matrix protein packing may modulate metabolite diffusion and, in turn, mitochondrial metabolism.     

重组耻垢分枝杆菌-Sj26GST疫苗的免疫保护作用研究

目的初步研究重组耻垢分枝杆菌疫苗rMc-Sj26GST(recombinant M.smegmatis mc2155-Sj26GST)对小鼠的免疫保护作用.方法用耻垢分枝杆菌重组疫苗rMc-Sj26GST免疫雄性BALB/c小鼠,检测小鼠淋巴细胞刺激指数(SI),腹腔巨噬细胞培养上清释放NO量,小鼠血清IFN-γ、IL-2的含量,并计数小鼠肝、肺活细菌数.结果小鼠脾淋巴细胞刺激指数(SI)为2.64±1.37,与Mc组(载体组)1.28±0.41相比,差异具有显著性;小鼠血清IFN-γ为196.43 pg*ml-1,与Mc组112.57 pg*ml-1相比,差异有显著性,较对照组高41%;小鼠血清中IL-2的浓度较对照组高.经rMc-Sj26GST疫苗免疫的小鼠受结核杆菌攻击后其肺、肝脏结核杆菌数较对照组少.结论耻垢分枝杆菌重组疫苗rMc-Sj26GST增强了小鼠细胞免疫功能,并使小鼠能抵抗结核杆菌的攻击.     

Mortality among female manual workers.

STUDY OBJECTIVE--The aim was to determine whether female manual workers have higher mortality than other women. DESIGN--This was a retrospective cohort study in which mortality was compared with that of the general female population. Main outcome measures were standardised mortality ratio (SMR) and 95% confidence intervals (CI). SETTING--Reykjavík region. PARTICIPANTS--Participants were 18,878 women, the cumulated members of a pension fund for manual workers between 1970 and 1986. MAIN RESULTS--A healthy worker effect was observed in the total cohort. The study was then restricted to those who had contributed to the pension fund any time after reaching 20 years of age, and a 10 year latency period was instituted. When analysing subcohorts by duration of employment the standardised mortality ratios for all causes of death and all cancers increased with longer employment time up to 10 years. However, the ratios were low in the group with over 10 years of employment. Those who began contributing to the fund in 1977 or later had higher mortality than those who began earlier. There was an excess of lung and bladder cancer in the total cohort and in all the subcohorts except in the group with over 10 years' employment. Mortality from accidents and suicides was in excess in all the groups. CONCLUSIONS--Mortality is high among some groups of female manual workers. A deficit was found among those with the longest employment. Differences in mortality have widened in recent years. An excess of suicides shows that women in this group have, for some reason, less will to live than other women.     

新的高强度高选择性阿片μ受体激动剂[11C]-羟甲芬太尼的合成

羟甲芬太尼(I)是一个新的高强度高选择性阿片μ受体激动剂。本文用cis-A-N-[1-(2-羟基-2-苯乙基)-3-甲基-4-哌啶基]-苯胺(II)或cis-N-[1-(苯甲酰甲基)-3-甲基-4-哌啶基]-苯胺(III)作为前体合成了[¹¹C]-羟甲芬太尼,以便用正电子发射断层扫描(PET)来观察μ受体。通过水解cis-A-羟甲芬太尼(I)和cis-N-[1-(苯甲酰甲基)-3-甲基-4-哌啶]-N-苯基丙酰胺(cis-IV)的4-N-丙酰基分别获得II和III。溴乙烷的格氏试剂与回旋加速器产生的[¹¹C]-二氧化碳反应后继而直接加入邻苯二甲酸二酰氯和2,6-二叔丁基吡啶生成同位素标记中间体[¹¹C]-丙酰氯。[¹¹C]-丙酰氯与OH-前体(II)反应后再经HPLC分离纯化直接得[¹¹C]-羟甲芬太尼;[¹¹C]-丙酰氯与酮-前体(III)反应后,再用硼氢化钠甲醇溶液处理,然后进行HPLC分离纯化得[¹¹C]-羟甲芬太尼。两种方法均可获得ll.1～14.8GBq/μmol的特异性放射化学纯[¹¹C]-羟甲芬太尼。总共耗时为40～50min(EOB)。     

Tuberculous wrist in the era of effective chemotherapy: an eleven-year experience.

BACKGROUND: The chronic course and non-specific clinical manifestations of tuberculous (TB) wrist often cause failure to make a timely diagnosis. OBJECTIVE: To better understand the rarely encountered TB wrist. PATIENTS AND METHODS: Retrospective review and analysis of cases of TB wrist between 1986 and 1997 in a medical centre in southern Taiwan. RESULTS: Thirty-seven cases (16 definitive, 13 probable and eight possible) of TB wrist (25 men, 12 women; mean age, 56.3 +/- 13.0 years) were found among a total of 4970 cases of tuberculosis. The most common presenting sign and symptom (mean duration 9.4 months) were local swelling and pain over the affected wrist. The mean white blood cell (WBC) count in peripheral blood was 7.04 x 10(9)/l, and the erythrocyte sediment rates (ESR) in seven of 31 patients who had ESR assayed were normal. Forty-six per cent of the patients had abnormal chest X-ray, and 35% had had previous manipulation of the affected wrist. CONCLUSION: Physicians should have a high index of suspicion for TB wrist among patients with chronic arthritis, even when their peripheral WBC count and ESR are normal. An abnormal chest X-ray and/or a history of previous manipulation of the affected wrist could be important clues for possible TB wrist.     

48例干扰素治疗慢性乙肝不良反应的观察及护理

重组α-2b干扰素是具有抗病毒和免疫调节作用的生物学调节活性物质。近年来，国外文献报道α-2b干扰素治疗慢性乙型病毒性肝炎(CAH—B)取得较好效果。特别是基因工程问世，使大量的高纯度干扰素应用于临床。现对48例慢性乙肝患者使用干扰素后的不良反应的观察与护理，报道如下。