In vitro differentiation of human adipose-derived adult stromal cells into neuron-like cells in hippocampal astrocyte conditioned medium

BACKGROUND: At present, researches on differentiating from human adipose-derived adult stromal cells (hADASC) to neuron-like cells are focus on inducing by artificial-synthetic compound solution; however, hippocampal astrocyte conditioned medium (HCAM) can induce in vitro differentiation from hADASC to neuron-like cells is still unclear. OBJECTIVE: To observe whether HCAM can induce in vitro differentiation from hADASC to neuron-like cells. DESIGN: Randomized control study. SETTING: Department of Neurology, Taixing People's Hospital; Central Laboratory, North China Coal Medical College. MATERIALS: Donor of adipose tissue was donated by female volunteers suffering from caesarean section in the department of obstetrics & gynecology in our hospital and aged 20－35 years. Adipose tissue was collected from subcutaneous tissue of abdomen during the operation. In addition, 8 male newborn Wistar rats within 24 hours with average body mass of 20 g were provided by Animal Institute of Chinese Academy of Medical Sciences. Rabbit-anti-human Nestin polyclonal antibody, rabbit-anti-human glial fibriliary acidic protein (GFAP) polyclonal antibody, rabbit-anti-human neuro-specific enolase polyclonal antibody and mouse-anti-human microtubal associated protein 2 (MAP-2) polyclonal antibody were provided by Wuhan Boster Company. METHODS: The experiment was carried out in the Central Laboratory of North China Coal Medical College from October 2004 to June 2005. hADASC was cultured with HCAM and its growth and morphological changes were observed under inverted phase contrast microscope. Immunocytochemistry, immunofluorescence and Western blotting were used to evaluate the expressions of Nestin, which was a specific sign of nerve precursor, neuro-specific enolase and MAP-2, which was a specific sign of nerve cell, and GFAP, which was a specific sign of neuroglial cells. MAIN OUTCOME MEASURES: Nestin, which was a specific sign of nerve precursor, neuro-specific enolase and MAP-2, which was a specific sign of nerve cell, and GFAP, which was a specific sign of neuroglial cells. RESULTS: On the 3rd day of culture, partial hADASC started deformation from slender shuttle-shape cells to neuron-like cells. It suggested that cells stretched out apophysis, which were mainly double-pole or multiple-pole cells. Five days later, immunohistochemical detection suggested that expression of Nestin (10.5±0.037) was found out in cells; meanwhile, expressions of GFAP (38.4±0.052) and neuro-specific enolase (NSE) (15.7±0.023) were also found out in cells; however, expression of MAP-2 was not observed. Western blot indicated that, 5 days after effect of HCAM, Nestin was found out in hADASC; meanwhile, expressions of GFAP and neuro-specific enolase were also found out; however, expression of MAP-2 was not observed. CONCLUSION: HCAM can induce the differentiation from hADASC to neuron-like cells in vitro.     

Whole-brain 3D perfusion MRI at 3.0 T using CASL with a separate labeling coil.

A variety of continuous and pulsed arterial spin labeling (ASL) perfusion MRI techniques have been demonstrated in recent years. One of the reasons these methods are still not routinely used is the limited extent of the imaging region. Of the ASL methods proposed to date, continuous ASL (CASL) with a separate labeling coil is particularly attractive for whole-brain studies at high fields. This approach can provide an increased signal-to-noise ratio (SNR) in perfusion images because there are no magnetization transfer (MT) effects, and lessen concerns regarding RF power deposition at high field because it uses a local labeling coil. In this work, we demonstrate CASL whole-brain quantitative perfusion imaging at 3.0 T using a combination of strategies: 3D volume acquisition, background tissue signal suppression, and a separate labeling coil. The results show that this approach can be used to acquire perfusion images in all brain regions with good sensitivity. Further, it is shown that the method can be performed safely on humans without exceeding the current RF power deposition limits. The current method can be extended to higher fields, and further improved by the use of multiple receiver coils and parallel imaging techniques to reduce scan time or provide increased resolution.     

Effects of human amniotic fluid on cartilage regeneration from free perichondrial grafts in rabbits.

After the chondrogenic potential of free grafts of perichondrium was shown in several experimental studies, perichondrium has been used to reconstruct cartilage tissue in various clinical situations. This study investigates the effects of human amniotic fluid on neochondrogenesis from free perichondrial grafts in a rabbit model. Since this fluid contains high concentrations of hyaluronic acid, hyaluronic acid-stimulating activator, growth factors, and extracellular matrix precursors during the second trimester, it may have a stimulating effect on neochondrogenesis. Perichondrial grafts, measuring 20 x 20 mm2 were obtained from the ears of 144 New Zealand young rabbits and were sutured over the paravertebral muscles. The rabbits were randomly divided into three groups with 48 rabbits per group. In group 1, 0.3 ml human amniotic fluid, and in group 2, 0.3 ml saline were injected underneath the perichondrial grafts. Group 3 formed the control group in which no treatment was given. Histologically, neochondrogenesis was evaluated in terms of cellular form and graft thickness at 2, 4, 6, and 8 weeks after surgery. In group 1, the mature cartilage was generated quickly and the cartilage plate in this group was significantly thick and extensive when compared with groups 2 and 3 at 8 weeks ( p<0.05 ANOVA). In conclusion, our study shows that human amniotic fluid enhances neochondrogenesis from free perichondrial grafts. The rich content of hyaluronic acid and growth factors possibly participate in this result.     

女性压力性尿失禁的微创治疗

目的探讨女性压力性尿失禁的发病机制和微创治疗方法。方法女性压力性尿失禁患者25例,经临床、尿动力学检查或膀胱尿道造影确诊,均为稳定性膀胱,无膀胱出口梗阻。采用无张力阴道吊带术(TVT),将吊带无张力置于尿道中段。结果手术时间平均(40±5)min;术中出血量平均(43±10)mL;术后平均留置导尿1.5 d;术后平均住院3 d。23例患者术后尿失禁症状均消失,其中2例患者术后出现尿潴留,留置导尿1周后好转,尿潴留症状消失,尿失禁未复发;2例尿失禁症状显著改善。结论TVT等微创术式因简单、微创、疗效好、并发症少等,目前是女性压力性尿失禁外科手术治疗的发展方向。     

多巴酚丁胺负荷超声心动图及组织多普勒成像技术在诊断冠心病中的临床应用

目的探讨多巴酚丁胺负荷超声心动图(DSE)和组织多普勒成像技术(TDI)在冠心病(CHD)诊断中的应用价值,以寻求一种敏感的、无创的CHD诊断方法。方法选择胸闷或胸痛怀疑CHD患者40例,应用TDI测量静息状态(Rest)下前壁中段收缩峰值速度(Vs),以及多巴酚丁胺峰值负荷状态(Peak)下前壁中段Vs,同时测量静息心率和负荷心率;所有入选对象均进行冠脉造影,并根据左前降支狭窄程度是否大于等于50%分为左前降支狭窄组(LST组)与左前降支非狭窄组(non-LST组)。分别比较Rest和Peak前壁中段LST组与non-LST组平均Vs差别等。结果Rest前壁中段LST组与non-LST组Vs差别无显著性意义;但Peak前壁中段LST组与non-LST组差别有显著性意义。结论DSE结合TDI是诊断冠心病有价值的定量分析方法。DSE在诊断CHD时的敏感性、准确性明显优于静息状态超声心动图。     

ULTRASTRUCTURAL INVESTIGATION ON THE ARTICULAR CARTILAGE FROM RABBIT KNEE JOINT AFTER IMMOBILIZATION IN CAST

The articular cartilages from the left knee of 30 rabbits immobilized in casts in functional position were studied under transmission electron microscope. The pathological changes which begin at early stage of immobilization included both degeneration and repair of cartilage. The striking morphological features observed were: (1) most chondrocytes in various layers of cartilage underwent progresive degeneration; (2) the concomitant abnormal proliferation of some chondrocytes happened in a brief period. The changes in mechanical stress caused by immobilization affected the nutrition of cartilage and finally led to its reactive structural reformation.     

大肠癌旁粘膜细胞的PCNA和AgNORs表达观察

用增殖细胞核抗原（ＰＣＮＡ）和核仁组成区嗜银蛋白（ＡｇＮＯＲｓ）对３５例大肠癌旁粘膜进行细胞增殖表达与观察。结果表明：癌旁粘膜的ＰＣＮＡ标记指数增多，标记增强，排列紊乱。ＡｇＮＯＲｓ在癌旁粘膜中，颗粒增多、增大，形态多样，０￣１ｃｍ和２￣３ｃｍ组的标记指数与４￣５ｃｍ组的标记指数相比较，有显著或非常显著差异（Ｐ〈０．０５或Ｐ〈０．０１）。在ＤｕｋｅＡ、Ｂ期中，细胞标记指数增高增强，与癌的发生和患者     

Electron microscopical demonstration of horseradish peroxidase by use of tetramethylbenzidine as chromogen and sodium tungstate as stabilizer (TMB-ST method): a tracing method with high sensitivity and well preserved ultrastructural tissue.

Until now methods using tetramethylbenzidine (TMB) for electron microscopy (TMB-EM methods) are all unable to provide a maximum demonstration of transported horseradish peroxidase (HRP) while maintaining good ultrastructural tissue preservation. In order to solve this problem, we have attempted to adapt a newly developed, highly sensitive TMB method using sodium tungstate (ST) as the stabilizer (TMB-ST method) for HRP electron microscopic retrograde and anterograde fiber tracing. The present study shows that the TMB-ST method combined with diaminobenzidine-cobalt (DAB-Co) is more sensitive than existing TMB-EM methods and that ultrastructural details are well preserved with this combined method. The resultant reaction product complex after osmication is stable and is observed as characteristic crystal-like structures which are extremely electron dense and often aggregated into clumps. In contrast, the TMB-ST method without the DAB-Co step frequently produces a moderate electron-dense reaction product. Therefore, we recommend the TMB-ST method combined with DAB-Co for HRP electron microscopy.     

静脉内毒瘾者HIV感染的转归及影响因素

对ＨＣＶ血清标记阳性的静脉内毒瘤者进行半年和１５个月随访，并调查影响转归的部分因素。结果显示，静脉内毒瘾者抗－ＨＣＶ自然转阴率分别为１１．０／和３３．３％。单因素和多因素分析均提示转阴率以婚姻状况良好的已婚者较离婚者高、人均年收入高者较低者高，说明婚姻状况和人们的经济水平对毒瘾者抗－ＨＣＶ转阴有一定的影响。     

氨氯地平与尼群地平治疗高血压的多中心平行对照研究

采用多中心、随机、单盲、组间、平行对照的方法对488例原发性高血压患者进行氨氯地平或尼群地平4周治疗观察，显示氨氯地平的降压有效率为90．4％，较尼群地平显著为高。不良反应较少，安全性好。每日只需服药1次，剂量5～10mg，方便病人。