柴油机排出颗粒物对小鼠骨髓细胞微核率的影响

对柴油机排出的颗粒物（ＤＥＰ）诱发小鼠骨髓细胞微核进行了研究，结果表明柴油机排出颗粒物（ＤＥＰ）在体内可产生明显的毒性作用。其提取物的剂量在８０－４００ｍｇ／ｋｇ之间，微核率由９．２‰升至３４．７‰，呈现明显的剂量－反应关系。各实验剂量组与对照组比较，均具极显著性差异（Ｐ＜０．０１）。这种遗传毒性作用可能是造成该颗粒致癌性和致突变性的机制之一     

超声波对小鼠骨髓细胞微核率的影响

目的：研究超声波对小鼠骨髓细胞微核率的影响。方法：用诊断级超声波幅射小鼠,测定小鼠骨髓嗜多染红细胞微核的出现率。结果：经超声波辐射的小鼠的细胞微核率与阳性对照组比较有显著性差异（Ｐ＜ ０．０１）,与阴性对照组比较无差异（Ｐ＞ ０．０５）。结论：受幅射的小鼠骨髓嗜多染红细胞微核率无显著增加,诊断级超声波不会致突变。     

生物玻璃-纳米羟基磷灰石梯度涂层的生物相容性研究

目的对生物玻璃-纳米羟基磷灰石(BG-nHA)梯度涂层的生物相容性作初步的评价。方法用低温烧结法在钛合金表面制备生物玻璃纳米羟基磷灰石梯度涂层,利用L929细胞检测梯度涂层材料的细胞毒性。取体外分离培养扩增的人骨髓基质干细胞(hBMSC),接种于涂层材料上,通过绿色荧光蛋白染色、扫描电镜和MTT法观察细胞的黏附和生长情况。结果生物玻璃-纳米羟基磷灰石梯度涂层的细胞毒性分级为1级,人骨髓基质干细胞可以在涂层材料表面黏附和生长。结论生物玻璃-纳米羟基磷灰石梯度涂层有良好的生物相容性,具有潜在的临床应用前景。     

生物玻璃-纳米羟基磷灰石梯度涂层的生物相容性研究

目的 对生物玻璃-纳米羟基磷灰石(BG-nHA)梯度涂层的生物相容性作初步的评价.方法 用低温烧结法在钛合金表面制备生物玻璃纳米羟基磷灰石梯度涂层,利用L929细胞检测梯度涂层材料的细胞毒性.取体外分离培养扩增的人骨髓基质干细胞(hBMSC),接种于涂层材料上,通过绿色荧光蛋白染色、扫描电镜和MTT法观察细胞的黏附和生长情况.结果 生物玻璃-纳米羟基磷灰石梯度涂层的细胞毒性分级为1级,人骨髓基质干细胞可以在涂层材料表面黏附和生长.结论 生物玻璃-纳米羟基磷灰石梯度涂层有良好的生物相容性,具有潜在的临床应用前景.     

洛美沙星对小鼠骨髓嗜多染红细胞微核的影响

本文研究了洛美沙星对NIH小鼠骨髓嗜多染红细胞微核的影响。结果表明,环磷酰胺(150mg/kg)组小鼠骨髓嗜多染红细胞微核率为65.4±11.56‰,与不给药对照组(3.0±0.82‰)相比明显增高(P<0.001)。而洛美沙星不论是单次给药各剂量组还是多次给药组的微核细胞出现率与对照组相比无显著性差异(P>0.05)。结果提示洛美沙星对小鼠骨髓细胞遗传物质无致突变作用。     

中药商陆诱发小鼠骨髓细胞微核率的实验研究

目的：通过本实验研究探讨孕妇禁忌中药商陆（Phytolacca acinosa）对小鼠骨髓嗜多染红细胞微核频率的影响。方法：先用体重18－22g的雌性昆明小鼠,随机分为10组,即阴性（灌胃生理盐水）、阳性（腹腔注射环磷酰胺）对照组、各实验组用生药商陆及醋制商陆煎液分别以四个剂量组（I、Ⅱ、Ⅲ、Ⅳ）每天灌胃1次,5天后取骨髓观察嗜多染红细胞的微核率。结果：两种商陆水煎剂的I、Ⅱ剂量组诱发小鼠微核率虽有数目变化,但无统计学意义,且与阴性对照组相比;P＞0．05,与阳性对照组相比均有显著差异,P＜0．01。而Ⅲ、Ⅳ剂量组诱发微核率与阴性对照组相比：P＜0．01,与阳性对照组相比无显著差异。讨论：本实验结果提示孕妇禁忌中药商陆诱发小鼠微核率具有明显的剂量效应关系。因此,在临床应用药时,特别是育龄妇女更应慎重。     

巴豆对小鼠骨髓及胚胎肝细胞微核率的影响

目的　探讨孕妇禁忌中药巴豆水提液对小鼠骨髓细胞及胚胎鼠肝细胞微核率的影响。方法　采用小鼠骨髓嗜多染红细胞微核 (MN)实验法与小鼠胚胎肝转移微核实验法。结果　当小鼠用药剂量在 1g/kg、 5g/kg时微核率与阴性对照组无明显差异 ,10g/kg时诱发骨髓MN率与阴性对照组比较有显著差异 (P <0 0 1)。而在孕鼠用药 1g/kg、 5g/kg、 10g/kg各剂量组时均可诱发鼠胎肝细胞微核率增高。各组与阴性对照组比较都有显著性差异 (P <0 0 1)。结论　实验表明巴豆水提液在同等剂量作用下诱发胚胎鼠肝细胞微核率明显高于成年鼠骨髓细胞 ,此药可以通过胎盘屏障 ,对胎鼠具有更为显著的致遗传物质损伤作用。     

生物玻璃-纳米羟基磷灰石梯度涂层的制备及检测

目的研究生物玻璃-纳米羟基磷灰石梯度涂层(BG-nHA)的表征以及涂层与钛合金(Ti-6Al-4V)之间的结合能力。方法用低温烧结法在钛合金表面制备生物玻璃-纳米羟基磷灰石梯度涂层,通过SEM、能谱分析仪(EDS)、X射线衍射仪(XRD)、红外傅利叶变换分析仪(FITR)等对涂层表征进行检测,材料在Tris-HCL缓冲液中行浸泡试验,依据ISO13779-4:2002标准用抗拉试验测试梯度涂层在浸泡前后与金属基质之间的结合强度,同时测定涂层材料释放的离子和丢失重量。结果电镜结果显示梯度涂层呈多孔状,表面分布均匀的杆状纳米羟基磷灰石晶体,基底部与钛合金形成致密结合,未发现细微的裂纹。抗拉试验显示梯度涂层与钛合金结合强度达到(39.7±4.4)MPa。随着浸泡时间的延长,涂层材料的重量有部分丢失,释放的离子逐渐增加。结论采用低温烧结法可以制备生物玻璃-纳米羟基磷灰石梯度涂层,这种加入生物活性玻璃的梯度涂层可以加强涂层与钛合金的结合能力。     

钛合金表面新型生物玻璃/羟基磷灰石涂层的体内动物实验

目的为促进钛合金植入体与骨的结合,在其表面制备了生物玻璃/羟基磷灰石复合涂层,并植入兔子股骨内进行动物试验,采用等离子喷涂羟基磷灰石涂层和未涂层的Ti6Al4V合金作为对照。方法种植到期的植入体取出后进行组织学切片,采用品红-苦味酸染色后进行组织学观察,采用SEM高倍观察种植体与骨的结合界面,并对骨接触率和凹槽内骨长入量进行了统计分析和比较。结果三种植入体都具有良好的生物相容性。Ti6Al4V合金与骨之间是一种形态固定,而生物玻璃/羟基磷灰石涂层、等离子喷涂羟基磷灰石涂层可与骨形成骨键合。生物玻璃/羟基磷灰石涂层在植入期间与基体没有脱落,同时其与骨的接触率和凹槽内骨长入量要明显高于其余两个植入体,显示出促进骨生长的作用。结论由于具有良好的生物相容性和促进新骨生长的能力,生物玻璃/羟基磷灰石涂层可加快植入体与骨的愈合速度,在骨替代修复方面显示出优势和广阔的应用前景。     

钛合金表面新型生物玻璃/纳米羟基磷灰石涂层的细胞相容性评价

目的为检验前期制备的新型生物玻璃／纳米羟基磷灰石涂层的细胞相容性。方法在本实验中，采用L929成纤维细胞在涂层浸提液中的培养，检测了涂层的细胞毒性，采用人体骨髓基质干细胞存涂层上的培养，检测了涂层对人体骨髓基质下细胞增殖和代谢的影响。结果低于浓度的涂层浸提液（〈10％）埘L929细胞增殖具有促进作用，而高浓度的涂层浸提液（〉50％）对L929细胞的增殖具有抑制作用，其中100％的涂层浸提液对L929细胞的增殖抑制比例为20.9％，在细胞毒性分级中处于合格范围内。结论在培养早期，人体骨髓基质干细胞在涂层表面的增殖要优于对照组，涂层显示出良好的细胞相容性。由于合格的细胞毒性和良好的细胞相容性，该涂层的钛合金具有作为骨植入物的应用潜力。     

IL-3对小鼠骨髓来源树突状细胞分化发育的影响

比较 GM- CSF IL- 4与 IL- 3 IL- 4两种方法培养制备的树突状细胞 (Dendritic cell,DC)在形态、产量、免疫表型和抗原摄取能力方面的差异。用 GM- CSF IL- 4和 IL- 3 IL- 4分别诱导小鼠骨髓来源的前体细胞分化为成熟树突状细胞 ,通过相差显微镜、扫描电镜、激光共聚焦扫描显微镜进行形态观察 ,流式细胞术分析细胞免疫表型和摄取抗原能力。结果表明 ,IL- 3 IL- 4诱导的 DC产量高 ,细胞纯度好 ,形态上与 GM- CSF IL- 4诱导的DC类似 ;细胞免疫表型显示高表达 MHC 类分子 ,但低表达或不表达 CD80、CD86 ;IL- 3 IL- 4诱导的 DC具有强大的摄取抗原能力。 IL- 3替代 GM- CSF可诱导低表达共刺激分子的耐受型 DC     

红景天苷对骨髓抑制贫血小鼠骨髓基质金属蛋白酶的影响

本研究用免疫组织化学方法和酶谱电泳法观察了红景天苷对骨髓抑制贫血小鼠骨髓基质金属蛋白酶-2和-9（MMP-2、MMP-9）的影响，并探讨其在造血调控中的可能作用。免疫组化结果显示，各组小鼠骨髓细胞中均检测到MMP-2和MMP-9表达。与对照组相比，模型组及高、中、低剂量红景天苷组骨髓细胞中MMP-2、MMP-9蛋白表达明显增强。在造模后第4d，中剂量红景天苷组中MMP-2和MMP-9表达最强。在8d，分别是低剂量和中剂量红景天苷组中MMP-2和MMP-9表达最强。酶谱电泳结果显示，在对照组骨髓中可检测到66kD（MMP-2的酶原形式，proMMP-2）、62kD（MMP-2的活性形式，MMP-2）、86kD（MMP-9活性形式，MMP-9）和94kD（MMP-9的酶原形式，proMMP-9）4条明胶酶的活性带，其中MMP-9的活性最强。造模后骨髓造血微环境中明胶酶的活性明显降低，不同剂量的红景天苷均能明显促进proMMP-9和MMP-9的活性增强，使proMMP-2的活性减弱。结果表明，红景天苷可能通过促进骨髓细胞中基质金属蛋白酶表达增强、骨髓造血微环境中基质金属蛋白酶活性升高，进而导致ECM中或基质细胞膜上细胞因子的释放、骨髓微血管损伤的修复以及HSCs增殖、迁移和分化能力的增加来促进骨髓造血功能的恢复。     

Counts of Stromal Precursor Cells in the Bone Marrow and Heterotopic Bone Marrow Transplants from Mice Immunized with Group A Streptococcus Antigens during Different Periods after Immunization

The counts of stromal precursor cells in bone marrow transplants obtained from animals 2 months after their immunization with killed type 5 group A streptococcus vaccine drop almost 3-fold in comparison with transplants from normal donors. Six months after donor immunization, the count of stromal precursor cells in the transplants reaches the normal level. The count of stromal precursor cells in bone marrow transplants from normal mice transplanted to recipients 6 months after their immunization with killed streptococcus vaccine also virtually did not change in comparison with the counts in bone marrow transplants from normal donors transplanted to normal recipients. The weight and size of bone capsules of 6.5-month bone marrow transplants in intact recipients after transplantation from donors immunized 2 months before with killed type 5 group A streptococcus vaccine was 3-fold lower than in bone marrow transplants collected from intact donors. The content of stromal precursor cells in the femoral bone marrow of animals immunized with killed streptococcus vaccine was 2.5 time higher in comparison with the parameter in the femoral bone marrow of normal mice even 8 months after immunization. The results indicate a significant long-acting effect of streptococcal antigens on the bone marrow stromal tissue, specifically, on its osteogenesis potential. Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 147, No. 1, pp. 78-81, January, 2009     

Observations on Fixation and Staining of Bone Marrow Biopsies

Abstract

Ten bone marrow (BM) biopsies were taken from the iliac crest of six autopsy cases. The tissues were fured in Zenker's fixative for intervals between one and 18 hours with and without decalcification. Ultrasonic bombardment was used for one and two hours. The same procedures were also used following 18 hours fixation in 10% neutral buffered formalin. Coded slides were double blind evaluated for section quality and cytologic detail after staining with H & E, three Giemsa variants, and Gomori's iron stain.

Paraffin processed BM biopsies were optimally prepared by fixation in Zenker's fluid for 4-18 hours without decalcification. Prior formalin fixation did not influence the consistent brilliance of Giemsa staining with the preferred Sheehan Wright Giemsa stain. Overnight Zenker fixation with or without prior formalin exposure and the absence of decalcification is recommended for the routine preparation of BM biopsies for paraffin processing. (The J of Histotechnol 11:145, 1988.)     

Effect of Disturbed Plasma Oxidant Homeostasis on Hemopoietic Differentiation of Polypotent Bone Marrow Stem Cells in Mice Treated with Dipyridamole

The choice of hemopoietic differentiation by polypotent bone marrow stem cells was studied by morphological and chemiluminescent methods. This choice occurred on day 7 after heterotopic bone marrow transplantation and dipyridamole administration, under the effect of oxidative homeostasis imbalance in the plasma. Dipyridamole increased total antioxidant activity and 10-fold reduced the intensity of free radical oxidation. Disaggregation therapy with dipyridamole reduced platelet count in the peripheral blood and decreased total plasma antioxidant activity.     

白血病患者骨髓细胞CFU-GM培养及药敏试验分析

目的　研究不同抗白血病药物对白血病病人个体的敏感性。方法　采用悬浮式培养方式 (CFU GM) ,取白血病患者的骨髓细胞进行培养 ,加入多种抗白血病药物 ,观察白血病细胞对药物的敏感性 ,与空白对照组白血病细胞从形态学方面做比较观察。结果　不同抗白血病药物对不同白血病病人个体的敏感性存在差异 ;对药物敏感的白血病细胞形态结构出现明显改变或死亡。结论　该方法可作为判断患者对药物是否敏感的重要指标 ,为临床治疗提供参考依据。     

Abnormal Hemopoiesis in Long-Term Bone Marrow Culture from Tumor Necrosis Factor-Deficient Mice

We studied hemopoiesis in mice deficient by the tumor necrosis factor gene. The total number of cells in long-term bone marrow cultures from these mice 2-fold surpassed that in wild-type and tumor necrosis factor p55 receptor-deficient animals. Increased cell production was related to the absence of tumor necrosis factor expression by hemopoietic precursors. The total cell production by explanted hemopoietic cells from tumor necrosis factor-deficient mice did not depend on the genotype of irradiated stromal sublayer in long-term cell cultures from wild-type mice and animals deficient by tumor necrosis factor or p55 receptor. These results suggest that tumor necrosis factor, but not its p55 receptor, is involved in transduction of signals regulating production of cultured cells. Tumor necrosis factor probably regulates hemopoiesis in long-term bone marrow cultures by initiating apoptosis of hemopoietic cells or inhibiting cell proliferation. Increased cell production probably attests to the absence of one or both effects.     

Counts of Stromal Precursor Cells in Heterotopic Bone Marrow Transplants in Mice Immunized with Group A Streptococcus Antigens

The count of stromal precursor cells in bone marrow transplants from CBA mice, transplanted to animals immunized with killed type 5 group A streptococcus vaccine, decreased 4.5–6.5 times (depending on the transplant age) in comparison with the grafts transplanted to normal recipients. The counts of stromal precursor cells in 1.5–3-month bone marrow transplants from animals immunized with killed streptococcal vaccine transplanted to normal mice were virtually the same, while in 7-month transplants they decreased 2-fold in comparison with their counts in bone marrow transplants from normal CBA mice transplanted to normal animals. The content of stromal precursor cells in the femoral bone marrow of animals immunized with killed streptococcal vaccine was appreciably (3.5 times) higher than in the bone marrow of normal mice. The results attest to an appreciable effect of streptococcal antigens on the bone marrow stromal tissue and suggest that not all stromal precursor cells, whose count increases after injection of antigens, are responsible for transplantability of the stromal tissue in case of its heterotopic transplantation. __________ Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 140, No. 7, pp. 77–80, July, 2005     

免疫毒素用于自体骨髓移植治疗白血病的实验研究及初步临床

目前自体骨髓移植被越来越多地用于白血病治疗,然而在缓解期从病人抽取的骨髓细胞可能含有残留的肿瘤细胞,当大剂量放、化疗后回输给病人,将来有可能引起白血病复发。现在有很多方法可以用来去除这些瘤细胞,免疫毒素是安全有效的方法之一。本文报道了由抗CD2、CD5和CD8单抗与蓖麻毒素组成的抗T免疫毒素,抗CD9、CD10单抗与蓖麻毒素组成的抗C-ALL免疫毒素在体外对靶细胞的特异性细胞毒作用,及用于临床净化骨髓的初步结果。