慢性粒细胞白血病骨髓细胞体外培养净化作用实验研究

目的 :采用 FISH方法直接在干细胞水平对慢性粒细胞白血病 (CML )患者自体骨髓体外培养前后的间期细胞进行 bcr/abl融合基因检测 ,从而探讨 CML 骨髓细胞体外培养对自体骨髓移植物的净化作用。方法 :分离初治的慢性期 Ph+ CML 7例骨髓单个核细胞 (MNCs) ,体外培养 10 d,用免疫磁珠 (MACS)富集培养前后的 CD34+ 细胞 ,通过流式细胞仪检测培养前后 MNCs中 CD34+ 干细胞比例 ,然后用 FISH方法检测其中 bcr/abl融合基因 ,同时分别用正常人细胞及 K5 6 2细胞株作阴性及阳性对照。结果 :(1) 7例患者骨髓细胞培养前后 CD34+ 细胞中 bcr/abl融合基因平均检出率分别为 85 .3%± 4 .9%和 78%± 5 .1% ,有统计学意义 (P<0 .0 5 ) ,(2 )培养前培养体系中 CD34+ 细胞数量为 (6 .0 6 0± 1.5 6 4 )× 10 5个 ,培养后体系中的 CD34+细胞数量为 (5 .974± 1.4 2 4 )× 10 5个 ,两者无明显差异 (P>0 .0 5 )。 (3)在对照组中假阳性率为 2 .5 % ,假阴性率为 2 %。结论 :自体骨髓细胞体外培养对 CML 病人的骨髓肿瘤细胞有一定程度的净化作用 ;FISH技术直接在干细胞水平检测 bcr/abl融合基因 ,且能定量分析 ,较传统方法更适合对骨髓净化进行评价 ,为骨髓净化提供了一种更方便、可靠的评定方法     

颈前路植骨钢板内固定治疗创伤性枢椎前滑移

目的 评价C2、C3椎体间植骨、钢板内固定治疗创伤性枢椎前滑移的临床价值。方法 8例创伤性枢椎前滑移患行颈前路手术复位、椎间盘切除减压、自体髂骨植骨、钢板内固定术，平均随访1年，观察患术后颈椎生理高度、曲度重建和颈椎稳定性、运动情况。结果 8例患均获得完全的枢椎复位，C2、C3椎体在术后16周达到骨性融合，颈椎生理高度、曲度得以重建，旋转、屈伸功能良好，无钢板螺钉并发症。结论 颈前路钢板内固定是治疗创伤性枢椎前滑移的有效方法。     

人Heparanase基因真核和原核表达载体的构建及融合蛋白的表达

目的：构建人Heparanase基因的真核、原核表达载体，大肠杆菌表达其融合蛋白．方法：采用反转录．聚合酶链反应从人肝癌细胞株HepG2cDNA中，分别扩增出Heparanase编码基因，用限制性内切酶BamHI消化后，插入真核表达载体pcDNA3．1中，经酶切鉴定与测序证实后，连接成包括完整的人Heparanase基因ORF区的真核表达载体，以亚克隆法构建于原核表达载体pRSET的相应酶切位点，转化大肠杆菌BL21菌株，异丙基β-D硫代半乳糖苷(IPTG)诱导产生融合蛋白．结果：构建的人Heparanase基因表达载体经序列测定证实，与GenBank登录结果完全一致；双酶切鉴定证实，克隆基因正确插入载体pcDNA3．1及pRSET；SDS-PAGE证实融合蛋白表达成功．结论：成功构建了人Heparanase基因真核、原核表达载体，成功正确表达了6His／Heparanase融合蛋白     

On the Use of Paramagnetic Beads and Ferrofluids to Assess and Eliminate the Leukocytic Contribution to Oxygen Radical Generation by Human Sperm Suspensions

PROBLEM: To develop a methodology to determine a) the leukocytic contribution to reactive oxygen species generation by human sperm suspensions and b) the therapeutic value of removing these cellular contaminants. METHODS: Leukocytes were removed with paramagnetic beads or colloidal ferrofluids coated with anti-CD45 antibody. The sperm suspensions were monitored for oxidant generation by chemiluminescence, leukocyte contamination by immunocytochemistry, and fertilizing potential using zona-free hamster oocytes. RESULTS: Percoll®-prepared human sperm suspensions exhibited a competence for PMA-induced reactive oxygen species generation which was significantly correlated with leukocyte contamination. However, the purified spermatozoa remaining after paramagnetic bead treatment, also demonstrated an intrinsic capacity for PMA-responsive reactive oxygen species generation and, freed from the oxidative stress created by the leukocytes, exhibited a significantly enhanced capacity for sperm-oocyte fusion. CONCLUSIONS: Although human spermatozoa can generate reactive oxygen species, sperm function is inhibited by the additional oxidative stress created by contaminating leukocytes. Removal of these cells with paramagnetic beads enhances fertilizing potential.     

核医学影像设备的发展与临床应用

γ相机和SPECT只能进行常规单光子显像，PET和双探头SPECT符合显像系统既能进行单光子显像，又能进行正电子符合显像，PET／CT系统的出现不仅提供高质量的衰减校正图像，保证了正电子显像校正数据的可靠性，而且能进行同机图像融合，提高了影像定位诊断的准确性。本文简要介绍了核医学影像设备发展历程，PET和PET／CT的原理以及在临床的应用。     

Classification of thoracic and lumbar spine fractures: problems of reproducibility

Reproducibility of fracture classification systems in general has been a matter of controversy. The reproducibility of spinal fracture classifications has not been sufficiently studied. We studied the inter-observer and intra-observer reproducibility of the Magerl (AO) classification using radiograms, CTs and MRIs of 53 patients. We compared this classification with the older and simpler Denis classification. Five observers classified the fractures, first using the radiograms and CTs and, 6 weeks later, with radiograms and MRIs. Three of the observers repeated the readings after 3 months. Three observers also classified the fractures according to Denis. Agreement was measured using Cohen's kappa test. The type (A, B, C) classification of the AO system was fairly reproducible with CTs. With MRI this was only moderate. Group subclassification of the types yielded higher kappa values, corresponding to substantial agreement. The agreement was, in general, better with the Denis classification, but the variance was higher due to the difficulty of finding proper categories for some injury patterns. Although the AO classification allows proper registration of all kinds of injury, the reproducibility, especially at the type level, is problematic. Use of MRI and better definition of the distinctive properties of the three different types may enhance the reproducibility of the scheme.     

Three antidepressants (amitriptyline,dothiepin, fluoxetine), with and without alcohol,compared with placebo on tests of psychomotor ability related to car driving

Eight female volunteers received acute doses of amitriptyline 50 mg (AMI), dothiepin 50 mg (DOT), fluoxetine 40 mg (FLU) or placebo both with and without a ‘social’ dose of alcohol (ALC) equivalent to 0·5 g/kg body weight absolute alcohol. Performance on a variety of tests of psychomotor ability and cognitive function (critical flicker fusion, choice reaction time, tracking, Maddox Wing and simulated car steering) were performed at 1·5 and 4 hours following treatment. AMI and DOT both with and without ALC impaired performance on a range of tests at either or both 1·5 and 4 hours, although the effects of AMI and AMI + ALC were more widespread and severe than those found with either DOT or DOT + ALC. FLU and FLU + ALC showed no evidence of impairment on any test at either the 1·5 or the 4 hours assessments. The results suggest that there are differences between the experimental substances, at the doses used, in their intrinsic potential for impairing aspects of psychomotor performance and cognitive function.     

Fusion and failure following anterior cervical plating with dynamic or rigid plates: 6-months results of a multi-centric, prospective, randomized, controlled study

Anterior cervical plate fixation is an approved surgical technique for cervical spine stabilization in the presence of anterior cervical instability. Rigid plate design with screws rigidly locked to the plate is widely used and is thought to provide a better fixation for the treated spinal segment than a dynamic design in which the screws may slide when the graft is settling. Recent biomechanical studies showed that dynamic anterior plates provide a better graft loading possibly leading to accelerated spinal fusion with a lower incidence of implant complications. This, however, was investigated in vitro and does not necessarily mean to be the case in vivo, as well. Thus, the two major aspects of this study were to compare the speed of bone fusion and the rate of implant complications using either rigid- or dynamic plates. The study design is prospective, randomized, controlled, and multi-centric, having been approved by respective ethic committees of all participating sites. One hundred and thirty-two patients were included in this study and randomly assigned to one of the two groups, both undergoing routine level-1- or level-2 anterior cervical discectomy with autograft fusion receiving either a dynamic plate with screws being locked in ap - position (ABC, Aesculap, Germany), or a rigid plate (CSLP, Synthes, Switzerland). Segmental mobility and implant complications were compared after 3- and 6 months, respectively. All measurements were performed by an independent radiologist. Mobility results after 6 months were available for 77 patients (43 ABC/34 CSLP). Mean segmental mobility for the ABC group was 1.7 mm at the time of discharge, 1.4 mm after 3 months, and 0.8 mm after 6 months. For the CSLP- group the measurements were 1.0, 1.8, and 1.7 mm, respectively. The differences of mean segmental mobility were statistically significant between both groups after 6 months (P = 0.02). Four patients of the CSLP-group demonstrated surgical hardware complications, whereas no implant complications were observed within the ABC-group (P = 0.0375). Dynamic plate designs provided a faster fusion of the cervical spine compared with rigid plate designs after prior spinal surgery. Moreover, the rate of implant complications was lower within the group of patients receiving a dynamic plate. These interim results refer to a follow-up period of 6 months after prior spinal surgery. Further investigations will be performed 2 years postoperatively.     

Disc replacement using Pro-Disc C versus fusion: a prospective randomised and controlled radiographic and clinical study

Anterior cervical discectomy and fusion (ACDF) may be considered to be the gold standard for treatment of symptomatic degenerative disc disease within the cervical spine. However, fusion of the segment may result in progressive degeneration of the adjacent segments. Therefore, dynamic stabilization procedures have been introduced. Among these, artificial disc replacement by disc prosthesis seems to be promising. However, to be so, segmental motion must be preserved. This, again, is very difficult to judge and has not yet been proven. The aim of the current study was to first analyse the segmental motion following artificial disc replacement using a disc prosthesis. A second aim was to compare both segmental motion as well as clinical result to the current gold standard (ACDF). This is a prospective controlled study. Twenty-five patients with cervical disc herniation were enrolled and assigned to either study group (receiving a disc prosthesis) or control group (receiving ACDF, using a cage with bone graft and an anterior plate.) Radiostereometric analysis was used to quantify intervertebral motion immediately as well as 3, 6, 12 and 24 weeks postoperatively. Further, clinical results were judged using visual analogue scale and neuro-examination. Cervical spine segmental motion decreased over time in the presence of disc prosthesis or ACDF. However, the loss of segmental motion is significantly higher in the ACDF group, when looked at 3, 6, 12 and 24 weeks after surgery. We observed significant pain reduction in neck and arm postoperatively, without significant difference between both groups (P > 0.05). Cervical spine disc prosthesis preserves cervical spine segmental motion within the first 6 months after surgery. The clinical results are the same when compared to the early results following ACDF.     

Radiologic long-term results after cervical vertebral interbody fusion with Polymethyl Methacrylat (PMMA)

Long-term results of cervical interbody fusion with PMMA were evaluated in a retrospective study. X-ray films of 83 patients were obtainable. Post-operative follow-up in this series was between 15 and 20 years. The results show that PMMA is engrafted after about 2 years. Stable vertebral interbody fusion is obtained in about 90% of cases. Development of malignoma was not observed. Resorptive bone alterations, which can be seen in about 2% of cases one to two years after operation are shown not to be progressive. This process heals and stable fusion develops.