补肾活血方药对慢性再生障碍性贫血患者骨髓细胞体外影响的实验研究

目的　研究补肾活血方药对慢性再生障碍性贫血 (CAA)患者骨髓细胞体外增殖的影响 ,探讨其治疗CAA的机制。方法　采用细胞培养 ,光镜下观察补肾活血中药作用于CAA患者骨髓细胞后的形态学 ;3 H TdR掺入和液体闪烁技术方法 ,检测补肾活血中药对CAA患者骨髓细胞的增殖变化。结果　加入补肾活血中药组 ,CAA患者骨髓细胞培养第 4 8小时见细胞丛明显增多 ,72h见大量集落形成 ;3 H TdR掺入法显示 ,补肾活血中药组骨髓细胞的增殖 ,以每孔 75g L最有效。在 5 0～ 10 0g L范围内 ,与对照组比较差异有显著性 (P <0 .0 1) ;2 5g L和 12 5g L组与对照组比较差异无显著性 (P <0 .0 5 )。结论　补肾活血方药在体外有促进CAA患者骨髓细胞增殖的作用 ,为补肾活血中药治疗CAA提供了实验与理论依据     

流式细胞术检测非小细胞肺癌患者骨髓微转移

目的　应用流式细胞术检测非小细胞肺癌患者骨髓中的微转移。方法　骨髓经Ficoll梯度离心分离单核细胞后,用细胞角蛋白 19(Cytokeratin, CK19)单克隆抗体染色,应用流式细胞仪检测。结果　检测了 96例肺癌患者骨髓,发现 25例患者骨髓中有微转移。20例肺良性病变患者、10例正常人骨髓均无微转移。肺癌患者阳性率与患者病理分期、分化程度和细胞组织学类型密切相关 (P<0. 05)。结论　应用流式细胞术检测非小细胞肺癌患者骨髓中的微转移,有助于肺癌的综合治疗。     

大鼠骨髓巨噬细胞的分离,纯化,培养以及鉴定

目的：为深入研究骨髓巨噬细胞的生物学功能，探索一种简便的骨髓巨噬细胞的分离，纯化和培养的方法。方法：分离获取大鼠骨髓细胞，在60％DMEM培养液，20％马血清，20％(v／v)L929培养上清的诱导条件下进行体外贴壁培养，6～7天时获得纯度较高的贴壁细胞。采用倒置显微镜下观察生活状态、Wrights染色光镜观察、电子显微镜观察检测贴壁细胞形态学；酸性磷酸酶及非特异性酯酶染色测定细胞内酶的表达；吞噬鸡红细胞及墨汁颗粒实验检测其吞噬功能；免疫细胞化学染色观察细胞表面标志等生物学技术鉴定培养的贴壁细胞性质。结果：获得高纯度的巨噬细胞，且具有良好的吞噬功能，并且具备巨噬细胞的形态特征，特有的水解酶类及特有的表面标志-CD68。结论：本法是一种简易实用的体外分离、纯化、培养骨髓巨噬细胞的方法。     

麦胚提取物对辐射损伤修复的实验研究

目的 研究麦胚提取物对小鼠辐射损伤修复的调节和保护作用。方法 采用辐射前或辐射同时饲喂一定量的麦胚提取物，观察小鼠经X射线一次性全身照射后的临床症状、30d存活率、骨髓微核率、外周血白细胞总数在不同时间的变化。结果 与对照组(单纯照射)比较，饲喂麦胚提取物可使小鼠的头面部皮肤、小肠黏膜、肾脏损伤症状得到明显改善；30d存活率为86．17％(P＜0．01)，提高存活率41．79％，保护系数为1．72；骨髓微核率4．62‰；比对照组(12．14‰)降低(P＜0．01)；外周血白细胞总数在7，13，20，30d均显回升(P＜0.05，P＜0．01，P＜0．01，P＜0．01)。结论 麦胚提取物对小鼠辐射损伤修复有一定程度的调节和保护作用，对于辅助肿瘤放射治疗具有重要意义。     

Initial experience with dynamic MR imaging in evaluation of normal bone marrow versus malignant bone marrow infiltrations in humans

The purpose of this study was (a) evaluation of dynamic contrast-enhanced MR imaging of normal bone marrow versus malignant bone marrow infiltrations in patients with proven B-cell-type chronic lymphocytic leukemia (B-CLL) and (b) correlation with the clinical stage according to Binet (stages A, B, C) and response to therapy. Bone marrow imaging of the lumbar spine, pelvis, and proximal femurs was performed at 1.5 T in 45 patients without known malignancy and in 30 patients with B-CLL. The differences between opposed-phase and in-phase dynamic gradient-echo sequences before and up to 10 minutes after intravenous application of .1 mmol/kg body weight of gadolinium-diethylenetriamine penta-acetic acid (Gd-DTPA) were evaluated in normal bone marrow. The contrast-enhancement patterns of normal and malignant bone marrow were compared using the opposed-phase dynamic gradient-echo sequence. Ten of the patients with bone marrow infiltrations (Binet stage C) additionally underwent MR imaging follow-up during therapy. Opposed-phase gradient echo sequences demonstrated a signal decrease of normal bone marrow, and in-phase gradient echo sequences demonstrated a signal increase of normal bone marrow after administration of Gd-DTPA. The dynamic signal intensity time courses differed significantly (P < .05) between Binet stages B and C and controls as well as among the three Binet stages of B-CLL. In the 10 patients followed during therapy, MR imaging sensitively demonstrated response (n = 6), nonresponse (n = 2), or relapse after initial response (n = 2). In out-of-phase imaging, both normal bone marrow and initial bone marrow infiltration in CLL stage Binet A show signal decrease after administration of contrast agent, whereas there is increase in signal intensity in higher-grade bone marrow infiltration in Binet stage B or C disease. The signal loss of normal bone marrow in out-of-phase imaging is a phase effect rather than a T2* effect. The differentiation of initial from higher-grade bone marrow infiltration on out-of-phase images relies solely on a shift in the fat/water ratio.     

吉林192Ir放射事故患者局部急性放射损伤病理改变

目的观察一例大剂量全身极不均匀照射急性放射损伤患者数处局部病理改变。方法病理常规切片染色，光镜检查。结果吸收剂量右下肢为３７３８Ｇｙ，左手掌为８３０Ｇｙ，于照射后第８天行右下肢、左前臂截肢术。右手、左膝照射剂量相对较小但难以准确估计，红斑、水肿、水疱、溃疡出现相对较晚，于第５５天行右手指、左膝清创植皮术。镜下：右小腿局部皮肤和皮肤附属器官、皮下组织和骨胳肌广泛地坏死和弥漫性出血，但真皮中的立毛肌尚存；左手指、手掌皮肤表皮细胞空泡变性、坏死，并形成大小不一的囊泡，致使表皮与真皮分离，真皮弥漫性出血、中性粒细胞浸润，汗腺上皮细胞变性、坏死，皮下组织内弥漫性出血，少数脂肪细胞坏死。胫骨、腓骨上端和股骨、桡骨、尺骨下端处骨髓腔内造血组织各系细胞均消失。左膝、右手拇指、食指、中指皮肤示急性放射性溃疡。结论大剂量照射导致的急性放射损伤造成大面积、深达肌肉的软组织坏死及骨髓造血细胞消失。     

体外构建组织工程心脏瓣膜动物实验初步研究

目的探讨应用去细胞猪主动脉支架(decellularized porcine aortic valve scaffold，APAVS)与兔骨髓干细胞(rabbit bone marrow stromal cells，RBMSCs)体外构建组织工程心脏瓣膜的可行性。方法采用去垢剂-核酸酶消化法处理，去除猪主动脉瓣细胞成分，并做去细胞前后的形态学检查和生物力学测定；在去细胞支架上种植兔骨髓干细胞，行形态学检查和免疫组化测定。结果光镜及电镜证实，猪主动脉瓣膜中的细胞成分可完全去除，获得完整无细胞的纤维网状支架；瓣叶去细胞前后的断裂强度和断裂伸长率无明显变化；种植的RBMSCs可在ACPAV表面形成一层连续的细胞层。结论种植RBMSCs于ACPAV上，可体外构造组织工程人工心脏瓣膜。     

Absence of donor-type major histocompatibility complex class I antigen-bearing microglia in the rat central nervous system of radiation bone marrow chimeras

Localization of bone marrow-originated cells in the central nervous system (CNS) of the rat was investigated by using bone marrow chimeras. In order to do this, Lewis rats which carry major histocompatibility complex (MHC) class I antigens haplotype 1 (RT1.Al) were reconstituted with (Lew X PVG)F1 (RT1.Al/c) bone marrow cells after lethal irradiation. Transferred bone marrow cells were detected by immunohistochemical staining using a monoclonal antibody, OX27, specific for haplotype c of rat MHC class I antigens (RT1.Ac). The spleen and thymus of chimeric rats were fully reconstituted with transferred F1 cells 4 weeks after bone marrow transplantation. At this stage, mononuclear cells in the subarachnoid space of the CNS expressed OX27 antigen indicating that they were of bone marrow origin. A few OX27-positive blood cells were scattered in the CNS parenchyma 4-12 weeks after reconstitution. Ramified microglia, however, remained OX27-negative. Bone marrow-derived microglia were not observed throughout the period of examination until 24 weeks. In addition, experimental allergic encephalomyelitis (EAE) was induced in chimeric rats in order to augment the expression of MHC class I antigens on microglia. Even under this condition, no OX27-positive microglia were observed. Taken together, ramified microglia might be of neuroectodermal origin and there is little possibility that the microglia are derived from the bone marrow. However, if the ramified microglia are derived from blood cells, the microglia may be expected to have characteristic cell kinetics from the following points: (1) the precursor cells of the microglia may enter the CNS only at the perinatal stage; and (2) even under the condition in which lymphocytes and macrophages enter the CNS as observed in EAE, the precursor cells of the microglia are not supplied from the blood.     

营养干预对骨髓移植病人抗氧化力及脂质过氧化的影响

目的探讨外源性补充抗氧化性维生素对骨髓移植病人抗氧化力及脂质过氧化的影响。方法将19名骨髓移植病人随机分为实验和对照组,实验组在预处理前给予维生素C(300 mg/d)和维生素E(600 mg/d),对照组不予补充,其余处理两者相同。测定移植前后两组病人的血浆总抗氧化力、丙二醛、全血谷胱甘肽过氧化物酶及红细胞超氧化物歧化酶水平。结果(1)移植前后实验组血浆总抗氧化力、谷胱甘肽过氧化物酶和红细胞超氧化物歧化酶值一直处于正常或正常值附近,对照组则低于正常值,两组相比有显著差异;(2)移植后实验组丙二醛值很快降至正常,对照组则一直处于较高水平,两组有显著差异。结论移植前外源性补充抗氧化性维生素可提高骨髓移植病人机体的抗氧化力,减少了脂质过氧化物的产生,有效缓解移植前大剂量放、化疗对病人机体造成的高氧化胁迫压力。     

骨髓间充质干细胞的分离培养及生物学特性研究

目的：通过体外细胞的培养方法将骨髓间充质干细胞由骨髓血中分离出来并加以纯化，进一步在体外培养条件下研究其增殖及生长特征，从而为探讨后续利用组织工程学方法修复肌肉骨骼系统组织缺损的可能性提供实验基础。方法：从骨髓血中提取间充质干细胞，体外培养扩增，再以相差显微镜、电镜观察并绘制生长曲线。结果：（1）通过离体培养，可以使体内环境下低丰度的骨髓间充质干细胞实现数目扩增。（2）体外培养条件下骨髓间充质干细胞具有成纤维细胞的生长特性。（3）体外单层培养条件下贴壁生长的骨髓间充质干细胞会出现“返祖现象”。（4）骨髓间充质干细胞内细胞器与其生物活性变化相一致。结论：骨髓间充质干细胞体外培养成功为今后利用自体间充质干细胞通过组织工程学方法修复肌肉骨骼系统的组织缺损提供功能细胞奠定了基础。