免疫磁珠分选骨髓衍生肝干细胞亚群c-Kit+lin-

目的:利用免疫磁性细胞分选系统分离纯化骨髓衍生肝干细胞亚群c-Kit lin-。方法:实验于2006-07/08在南方医科大学实验动物中心完成。6～8周龄的SPF级纯系BALB/C雄性小鼠10只,体质量18～20g。收集小鼠股骨骨髓细胞,利用免疫磁性细胞分选系统,通过两步法分选纯化c-Kit lin-:将获取的lin-细胞悬液8℃条件下1500r/min离心10min,弃上清,按80μL/107加入Buffer重悬细胞。按20μL/107加生物素抗体磁珠,混匀,4℃冰箱孵育15min,按1mL/107加入Buffer洗细胞1次,8℃条件下1500r/min离心10min,弃上清,按500μL/108加入Buffer重悬细胞。Buffer500μL润MS柱,悬液过柱后,Buffer500μL/次洗柱3次,柱子脱离磁场,加1mLBuffer,用配套柱塞推出柱中的c-Kit lin-细胞,收集到c-kit lin-细胞,细胞计数。取2.0×106个细胞分成10等份,流式细胞仪分析c-Kit lin-细胞纯度,计算回收率,评估纯化效率,苔盼兰染色检测纯化前后的细胞活力。计算活细胞的百分率。细胞纯度和细胞回收率的计算:细胞纯度=分离产物中的阳性细胞数/分离细胞的总细胞数×100%,细胞回收率=分离产物中的阳性细胞数/起始标本阳性细胞总数×100%。结果:10只小鼠均进入结果分析。利用免疫磁性细胞分选系统分选出的骨髓衍生肝干细胞亚群c-Kit lin-细胞纯度和回收率分别为(77.98±2.34)%,75.40%,纯化前后细胞活力不受影响。结论:免疫磁性细胞分选系统能有效分选骨髓衍生肝干细胞亚群c-Kit lin-,纯度和回收率高,且不影响细胞活力。     

Lysophosphatidylcholine‐induced cytotoxicity and protection by heparin in mouse brain bEND.3 endothelial cells

A pathological feature in atherosclerosis is the dysfunction and death of vascular endothelial cells (EC). Oxidized low‐density lipoprotein (LDL), known to accumulate in the atherosclerotic arterial walls, impairs endothelium‐dependent relaxation and causes EC apoptosis. A major bioactive ingredient of the oxidized LDL is lysophosphatidylcholine (LPC), which at higher concentrations causes apoptosis and necrosis in various EC. There is hitherto no report on LPC‐induced cytotoxicity in brain EC. In this work, we found that LPC caused cytosolic Ca²⁺ overload, mitochondrial membrane potential decrease, p38 activation, caspase 3 activation and eventually apoptotic death in mouse cerebral bEND.3 EC. In contrast to reported reactive oxygen species (ROS) generation by LPC in other EC, LPC did not trigger ROS formation in bEND.3 cells. Pharmacological inhibition of p38 alleviated LPC‐inflicted cell death. We examined whether heparin could be cytoprotective: although it could not suppress LPC‐triggered Ca²⁺ signal, p38 activation and mitochondrial membrane potential drop, it did suppress LPC‐induced caspase 3 activation and alleviate LPC‐inflicted cytotoxicity. Our data suggest LPC apoptotic death mechanisms in bEND.3 might involve mitochondrial membrane potential decrease and p38 activation. Heparin is protective against LPC cytotoxicity and might intervene steps between mitochondrial membrane potential drop/p38 activation and caspase 3 activation.     

针刺足三里和悬钟治疗缺血性脑卒中

目的:观察针刺足三里、悬钟2穴对缺血性脑卒中脑血管功能的影响,分析其可能的作用机制,并对临床疗效做出评价。方法:选择2004-11/2006-05湖北中医药高等专科学校附属古城医院针灸科、荆州市第五人民医院中医康复科、荆州市第三人民医院中医科3单位缺血性脑卒中患者合适病例160例,采用查随机数字表的方法,将其随机分为对照组和针刺组,各80例。对照组采用现代医学常规干预方法进行治疗:卧床,保持呼吸道通畅,预防感染,控制颅内压、血压,维持水电解质平衡。针刺组在此基础上加针刺足三里、悬钟2穴,采用慢速捻转进针法针刺,留针20～30min,每隔5min行针1次。1次/d。两组患者治疗30d。并以经颅多普勒检测观察缺血性脑卒中患者治疗前后脑血管舒缩反应能力、脑血流自动调节功能、大脑半球侧枝循环代偿功能的变化,同时以治疗前后神经功能缺损程度为指标评价其临床疗效。结果:160例病例全部进入结果分析。①针刺组与治疗前相比,脑血管舒缩反应能力明显加强,差异有显著性意义(t=2.97,P<0.05),且优于对照组(t=2.45,P<0.05)。②针刺组与治疗前相比,脑血流自动调节能力明显改善,差异有非常显著性意义(t=8.01,P<0.01),且优于对照组(t=7.67,P<0.05)。③针刺组与治疗前相比,大脑半球侧枝循环代偿功能得到加强,差异有显著性意义(t=3.15,P<0.05),且优于对照组(t=5.16,P<0.05)。④针刺组与治疗前相比,神经功能缺损积分明显降低,差异有非常显著性意义(t=4.83,P<0.01),且优于对照组(t=5.43,P<0.05)。结论:针刺足三里、悬钟2穴对缺血性脑卒中患者脑血管舒缩反应能力、脑血流自动调节功能、大脑半球侧枝循环代偿功能有明显改善作用,并能促进神经功能的恢复。     

自体髂骨植骨带锁钢板内固定材料置入并一期前后路减压治疗多节段脊髓型颈椎病12例:临床资料回顾分析

目的:严重的多节段脊髓型颈椎病单纯前路或后路手术都有其局限性。观察一期前后路联合手术并自体髂骨植骨及带锁钢板内固定材料置入在治疗多节段脊髓型颈椎病中的应用价值。方法:选择2004-11/2006-12本院12例多节段脊髓型颈椎病患者,均采用一期前后路减压、自体髂骨植骨融合、带锁钢板内固定联合手术。其中男9例,女性3例,年龄49～75岁;3节段受累9例,4节段受累3例(突出节段分布:C3～66例,C4～73例,C3～73例)。全部病例进行临床随访,患者均对本试验知情同意。采用mJOA评分标准对患者神经功能改善情况进行评定;术前颈椎侧位片测量,以D值(C4椎体后下缘到齿突后缘与C7椎体后下缘连线的垂直距离)评价颈椎(C2～7)弧度;根据颈椎伸屈动态侧位片C2和C7椎体后缘切线相交所成的夹角之和评价颈椎(C2～7)活动范围。主要以电话随访和问卷填写的方式,分别从神经功能改善情况、颈椎弧度、活动范围及术后并发症等进行随访观察。结果:①12例患者全部得到随访,术后随访时间6～28个月,平均(16±6)个月。②所有植骨均获得骨性愈合;疗效结果中优4例(33.3%);良6例(50%);无效2例(16.7%);颈椎D值术前(3.9±1.4)mm,术后即刻(8.5±1.7)mm,随访时(8.1±2.5)mm。术前与术后差异有显著性(P<0.01),术后与随访时差异无显著性(P=0.251);颈椎活动范围术前(36.3±4.0)°,随访时(10.6±2.7)°,与术前相比差异具有显著性(P<0.01)。③术后C5神经根麻痹1例,为感觉及运动混合型,8个月随访时,感觉功能恢复,肩关节外展肌力从术后Ⅱ级恢复至Ⅳ级;1例术后6个月出现"S"畸形而再次压迫脊髓,神经功能改善停滞,目前处于随访中。结论:一期前后路手术并自体髂骨植骨及带锁钢板内固定材料置入减压充分、彻底,而且前路手术能重建颈椎稳定性,恢复颈椎生理前凸和椎间高度,并且后路减压术又能预防相邻颈椎退变引起的脊髓继发的压迫。     

Functional characterization of mesenchymal stem cells labeled with a novel PVP‐coated superparamagnetic iron oxide

Magnetic resonance imaging of cells labeled with superparamagnetic iron oxide (SPIO) could be a valuable tool for tracking transplanted cells in living organisms. Human bone marrow‐derived mesenchymal stem cells (hBMMSC) were labeled with a novel polyvinyl pyrrolidone (PVP)‐coated SPIO. Prussian blue staining and electron microscopy revealed that almost all of the cells were efficiently labeled with PVP–SPIO nanoparticles. There were no signs of cytotoxicity, even at concentrations of up to 1600 µg Fe/ml of the nanoparticles, and the labeled cells were successfully visualized by in vitro cellular MRI. In addition, there was no significant alteration of the phenotype or the adipo/osteo/chondrogenic differentiation potential of the cells. This was in contrast to Feridex IV labeling that led to the inhibition of hBMMSC chondrogenesis. Following intramuscular injection in a rabbit hind limb ischemia model, the intercellular migration of the labeled cells toward the ablated site was clearly tracked through in vivo MRI. The localization of the transplanted cells observed by MRI correlated well with postmortem histological studies. These results demonstrate that the novel PVP–SPIO nanoparticles appear to be efficient MR contrast agents and may enable non‐invasive in vivo tracking of stem cells in experimental and clinical settings during cell therapy. Copyright © 2009 John Wiley & Sons, Ltd.     

Prevalence and time trend of intestinal metaplasia in Hong Kong

Background and Methods:  Upper endoscopy records from 1998 to 2003 were reviewed. The demographic data, endoscopic diagnosis, results of rapid urease test and the absence or presence of intestinal metaplasia (IM) in histology were reviewed, to evaluate the prevalence of IM and Helicobacter pylori (Hp) infection over time in Hong Kong.
Results:  Among 1805 endoscopies performed, 1751 had both rapid urease test and histology available. A significant drop in the prevalence of duodenal ulcers from 17.9% in 1998 to 9.8% in 2003 was found ( P  = 0.015). Prevalence of IM was 13.9%, 5.9% and 9.4% in Hp positive, Hp negative and overall respectively ( P  < 0.05). The prevalence of IM increased with age, and the patterns were similar amongst subjects in 1998–2000 and those in 2001–2003. There was progressive decrease in Hp prevalence from 58% in 1998 to 40% in 2001 ( P  = 0.014), but no further decrease was seen in 2002–3. There was no corresponding decrease in IM prevalence. Instead IM prevalence in 2002–2003 was significantly higher than the prevalence in previous few years ( P  = 0.04).
Conclusion:  The prevalence of IM did not change in the period from 1998 to 2003 despite a drop in the prevalence of Hp infection since 1994.     

Inhibitory effect of Carthamus tinctorius L. seed extracts on bone resorption mediated by tyrosine kinase,COX-2 (cyclooxygenase) and PG (prostaglandin) E2

Anti-bone resorption properties of the Korean herbal formulation, Honghwain (HHI; Carthamus tinctorius L. seed) was biochemically investigated. On processing bone metabolism, PGE2 accelerated production of IL-1beta in fetal mouse osteoblast and stimulated physiological activation substance, IL-1beta. The novel class of Src tyrosine kinase inhibitors, Herbimycin A (HERB) and HHI reduced COX-2 mRNA levels as well as PGE2 production induced by IL-1beta, TNF-alpha and IL-6. HHI inhibited in vitro and in vivo bone resorption by inhibition of phosphorylation of peptide substrates. HHI dose-dependently reduced the hypercalcemia induced in mice by IL-1beta and partly prevented bone loss and microarchitectural changes in young ovariectomized rats, showing that the protective effect on bone was exerted via the inhibition of bone resorption. These results indicate that the synergy between IL-beta, TNF-alpha, IL-6 on PGE2 production is due to an enhanced gene expression of COX-2 and that tyrosine kinase (s) are involved in the signal transduction of COX-2 in mouse calvarial osteoblasts. Thus, HHI as a possible Src family kinase inhibitor may be useful for the treatment of diseases associated with elevated bone loss.     

金龙胶囊配合辨证用药治疗中晚期恶性肿瘤40例

1临床资料2003-12/2004-10采用北京建生药业生产的金龙胶囊配合辨证用药治疗中晚期癌症40例.Karnofsky评分≥40分,预计生存期>2 mo.     

Parents' perceptions towards their child's use of inhaled medications for asthma therapy

Objective : Understanding patients' and their parents' perceptions towards asthma medication is important in developing strategies to ensure patient compliance. In this study, parents' perceptions towards their children's use of inhaled medication for asthma treatment were evaluated.
Methodology : A questionnaire was administered by interviewing parents of 210 asthmatic children attending the Department of Paediatrics, National University Hospital.
Results : Our results showed that 76/210 (36%) of parents either felt opposed to inhaler therapy and/or preferred oral medications. The main reasons for their reluctance to use inhalers were related to fear of dependence, side effects and overdosage, and the child's dislike for inhalers. A third of these parents felt that inhalers were only indicated for very severe asthma.
Conclusions : We conclude that a significant proportion of parents have reservations regarding the use of inhalers for the treatment of asthma. These factors should be taken into consideration when planning an effective asthma education programme.