New Stains for Blood and Bone Marrow Cells

Traditionally, blood and bone marrow cells have been identified based on their characteristic shapes and colors when stained with one of several panoptic stains including Wright's or Giemsa's. As questions arose regarding the origin of normal and leukemic cells, cytochemical stains were developed. These stains help identify cells on the basis of a distinctive metabolite or enzyme. As part of an ongoing tradition in which textile dyes are used for biological staining, several new stains have been applied to hematologic staining. These include C.I. basic blue 41, basic blue 141, basic blue 93, and an assymetrical polymethine dye. As additional cell-selective stains are developed, we can anticipate further improvements in our ability to identify normal and malignant hematopoietic cells.     

Comparison of Trypanosome Incidence in Blood and Bone Marrow from 79 Colorado Birds

Seventy-nine adult birds in 8 species were examined at autopsy for the presence of Trypansoma avium. Films of thoracic blood were stained in Giemsa and later searched for trypanosomes. Marrow was removed from femora and humeri, mixed with Ringer's solution, and examined direct for living flagellates. Sixty-four birds (81%) showed positive marrows; 5 (6%) had positive bloods. In the 15 birds with negative marrows, only 2 had trypanosomes in the blood. These data emphasize the advantage of marrow over blood examination in determining the incidence of avian trypanosomiasis and point up the excellence of fresh marrow examination in such studies.     

Microspectrophotometric Studies of Romanowsky Stained Blood Cells. IV. Maturation of Myeloid and Erythroid Cell Lines in Bone Marrow

A quantitative characterization has been made of azure B/eosin stained cells from bone marrow. Two cell lines were followed: the myeloid line (white cell blast, promyelocyte, neutrophilic myelocyte, neutrophilic metamyelocyte, neutrophilic hand, neutrophilic segmented) and the erythroid line (rubriblast, prorubricyte, rubricyte, metarubricyte, diffusely basophilic erythrocyte, erythrocyte). A consensus scheme was used to obtain the “true” classification of the cells. Cell types were characterized by three methods: absorbance spectra, dye binding, and chromaticities. Within both cell lines nuclear maturation is accompanied by an overall increase in peak absorbance with little shift in the position of the maximum. Generally, binding of azure B and eosin increases; azure B dimer/monomer ratios show a slight downward trend during maturation. Changes in chromaticities are to bluish purples of increasing saturation. Cytoplasmic changes accompanying maturation are much more striking than nuclear changes, and again the two cell lines show similarities. Generally, there is decreased binding of azure B during maturation. In the erythroid line, the Soret band of hemoglobin becomes increasingly prominent. Chromaticities change from bluish purples to purplish pinks, particularly in the erythroid line.     

Cytogenetic Analysis of Bone Marrow and Peripheral Blood Samples Stored in Fixative for Several Years

We have developed a method which improves the spreading of chromosomes and permits banding analysis of cytogenetic samples of bone marrow and unstimulated peripheral blood which have been stored in fixative for up to 15 years. Metaphase cells had been harvested as usual and stored in fixative (acetic acid:methanol 1:3) at -15 C. The procedure includes 4-5 changes of fixative (acetic acid:ethanol 1:1). Next, cells are dropped onto a chilled, wet slide. The back of the slide is then rinsed with 70% ethanol and dried by ignition. C-, G-, Q-, or R-banding patterns can now be obtained with these specimens. The procedure is useful for reinvestigation of cytogenetic samples that were obtained prior to the development of banding techniques.     

Bone Marrow Transplantation Concurrently Reconstitutes Donor Liver and Immune System across Host Species Barrier in Mice

Liver immunopathologic mechanisms during hepatotropic infection, malignant transformation, and autoimmunity are still unclear. Establishing a chimeric mouse with a reconstituted liver and immune system derived from a single donor across species is critical to study regional donor immune responses in recipient liver. Using a strain of mice deficient in tyrosine catabolic enzyme fumarylacetoacetate hydrolase (fah^-/-) and bone marrow transplantation (BMT), we reconstituted the donor''s hepatocytes and immune cells across host species barrier. Syngeneic, allogeneic or even xenogeneic rat BMT rescued most recipient fah^-/- mice against liver failure by donor BM-derived FAH⁺ hepatocytes. Importantly, immune system developed normally in chimeras, and the immune cells together with organ architecture were intact and functional. Thus, donor BM can across host species barrier and concurrently reconstitutes MHC-identical response between immune cells and hepatocytes, giving rise to a new simple and convenient small animal model to study donor''s liver immune response in mice.     

小鼠骨髓造血干细胞、外周血组成随年龄的变化趋势及其相关性分析

造血干细胞是具有自我更新能力并能分化为血液中各种血细胞组分的多能干细胞。近来研究显示,不同造血干细胞表面标志物标记的造血干细胞具有分化为不同血细胞的趋势,但是这种分化的内在关系仍不清楚。对小鼠CD34~-/Sca-1~+骨髓造血干细胞、外周血组成随小鼠年龄增长的变化情况进行了分析,结果显示:随着年龄的增长,骨髓中的CD34~-/Sca-1~+骨髓造血干细胞比率显著增加;而外周血各组分则随年龄变化呈现不同的趋势。对不同年龄段小鼠的骨髓造血干细胞及其他组分与外周血组分的同步分析发现,外周血中血小板密度变化趋势与CD34~-/Sca-1~+骨髓造血干细胞变化情况相关系数为0.804 8;外周血中淋巴细胞密度变化趋势与CD34~+/Sca-1~-骨髓细胞的变化情况相关系数为0.947 97;外周血中白细胞密度变化趋势与CD34~+/Sca-1~+骨髓细胞变化情况相关系数为0.763 1(大于0.9为极度相关,0.7到0.9为高度相关)。     

Dysfunction of Bone Marrow Vascular Niche in Acute Graft-Versus-Host Disease after MHC-Haploidentical Bone Marrow Transplantation

Acute graft-versus-host disease (aGvHD) is the most common complication of allogeneic hematopoietic stem cell transplantation (HSCT), which is often accompanied by impaired hematopoietic reconstitution. Sinusoidal endothelial cells (SECs) constitute bone marrow (BM) vascular niche that plays an important role in supporting self-renewal capacity and maintaining the stability of HSC pool. Here we provide evidences that vascular niche is a target of aGvHD in a major histocompatibility complex (MHC)–haploidentical matched murine HSCT model. The results demonstrated that hematopoietic cells derived from GvHD mice had the capacity to reconstitute hematopoiesis in healthy recipient mice. However, hematopoietic cells from healthy donor mice failed to reconstitute hematopoiesis in GvHD recipient mice, indicating that the BM niche was impaired by aGvHD in this model. We further demonstrated that SECs were markedly reduced in the BM of aGvHD mice. High level of Fas and caspase-3 expression and high rate of apoptosis were identified in SECs, indicating that SECs were destroyed by aGvHD in this murine HSCT model. Furthermore, high Fas ligand expression on engrafted donor CD4⁺, but not CD8⁺ T cells, and high level MHC-II but not MHC-I expression on SECs, suggested that SECs apoptosis was mediated by CD4⁺ donor T cells through the Fas/FasL pathway.     

骨髓单个核细胞抗体及外周血B淋巴细胞与IRP的相关性研究

采用流式细胞术双标法检测拟诊免疫相关性全血细胞减少症患者(A组)、非免疫相关性恶性血液病患者(B组)及正常人(正常对照组)的骨髓单个核细胞结合的自身抗体,同时检测B组、确诊免疫相关性全血细胞减少症(C组)及正常对照组外周血B淋巴细胞及CD5+B淋巴细胞比率;A组中16例骨髓造血细胞自身抗体阳性,阳性率88.88%;B组1例骨髓造血细胞自身抗体阳性,阳性率9.09%。C组外周血B淋巴细胞、CD5+B淋巴细胞比率显著高于B组及正常对照组(P均<0.05),而正常对照组外周血B淋巴细胞、CD5+B淋巴细胞显著高于B组(P均<0.05);IRP患者骨髓单个核细胞自身抗体表达显著增高,B淋巴细胞总数及CD5+B淋巴细胞数量显著增高可能是IRP发病的重要因素之一;利用流式细胞术检测骨髓造血细胞自身抗体及B淋巴细胞数可以为IRP提供科学可靠的依据,优于骨髓Coomb’s实验。     

Rate and Time of Dna Synthesis of Human Leukaemic Blasts In Bone Marrow and Peripheral Blood

Abstract. We have evaluated DNA synthesis rate (S rate) and time (T_s) and tritiated thymidine labelling index (LI) of peripheral blood (PB) and/or bone marrow (BM) leukaemic blasts (Bl) in nineteen cases of acute leukaemia (twelve non-lymphoblastic, AnLL, and seven lymphoblastic, ALL), in one case of non-Hodgkin's leukaemic lymphoma and in a case of plasma cell leukaemia.
The LI of PB-BI was significantly lower than that of BM-Bl (range 0.1-6.2% and 1.9-19.5%, respectively; P < 0.01). the S rate was higher for PB-Bl than for BM-Bl (range 3.5-11-3 and 2.5-9.5 mol × 10-¹⁸/min; P < 0.02) and the T^s of PB-Bl was shorter than that of BM-Bl (range 7.6-22.1 and 10.8-34.7 hr, respectively; P < 0.02). In eight cases where S rates of both BM-Bl and PB-Bl were available, a linear correlation ( r = 0.82; P < 0.01) was found between the two parameters. This suggests that the DNA synthetic rate is a property of the leukaemic cell line in individual patients and differs from case to case. It further indicates that the environmental influences on the DNA synthesis rate in BM or PB are always of the same order of magnitude. From the results of this study we speculate that the DNA synthesis rate of leukaemic blasts is slowed down in the BM by environmental factors such as cell density.     

共表达PDX1和BTC间充质干细胞对转分化胰岛素分泌细胞的影响

该文通过Tet调控下共表达PDX1与BTC的骨髓间充质干细胞系(PDX1~+BTC~+MSCs),探讨PDX1和BTC共表达对骨髓间充质干细胞分化为胰岛素分泌细胞(IPCs)的效率及成熟度的影响。采用两步法对PDX1~+BTC~+MSCs细胞系诱导分化成IPCs,第一步Dox诱导7天检测到Nestin、CK19表达;第二步再诱导7天后形成DTZ染色阳性的胰岛样结构,Ngn3、Nkx6.1 mRNA水平和PDX1、Insulin、Glucagon的蛋白表达阳性。分化后的IPCs在葡萄糖刺激下能产生胰岛素和C肽,但仍不能达到正常胰岛水平。提示利用Tet-On体系调控PDX1和BTC共表达对骨髓间充质干细胞进行修饰,能有效诱导骨髓间充质干细胞分化为胰岛素分泌细胞,但分化成熟度仍然与天然胰岛细胞功能存在差距。     

The Effect of Early Application of a Combined Therapy of Bone Marrow Mesenchymal Stem Cells and Platelet-Rich Plasma on Blood and Bone Parameters in Ovariectomized Rats

Journal of Evolutionary Biochemistry and Physiology - Osteoporosis is a common bone disorder with marked morbidity and mortality that occurs frequently in women after menopause. Inadequacy of...     

骨髓干细胞的可塑性研究进展

成体干细胞在体内特定的微环境或体外人工培养条件下具有极强的可塑性分化潜能,其主要功能是负责组织细胞的生理性更新和病理性修复．骨髓组织中包括产生所有成熟血细胞系的造血干细胞(HSCs)、多潜能成体祖细胞和能分化为骨、软骨、脂肪的间充质干细胞(MSCs),这些细胞时还有向造血和骨髓以外的其他类型的成熟细胞分化如神经、肌肉、皮肤、心、肝、肾、肺等分化的能力．对最近几年国内外关于骨髓干细胞可塑性的实验研究进展作简要综述．     

骨髓移植与造血干细胞研究

骨髓移植是目前治疗白血病等血液疾病最有效的方法。在经历了多年的发展并已取得巨大成就的今天,骨髓移植仍然面,临很多问题。造血干细胞研究必将解决骨髓移植中的诸多问题。重点对造骨髓移植的发展历史,所面临的问题以及造血干细胞研究的进展进行简要的概述。     

Cell Interaction Molecules Utilized in Bone Marrow

Many aspects of blood cell formation can now be modeled in culture and rapid progress is being made in understanding how blood cell precursors interact with unique components of their environment. This brief review considers some cell interaction molecules that may be important for controlling the position of cells within, as well as their egress from, bone marrow.     

流行性出血热病毒对健康人外周淋巴细胞和大白鼠骨髓细胞染色体影响的研究

用流行性出血热病毒(EHFV)A9株,滴度为TCID_(50)10~(-5)/0.1ml,加入10名健康人外周血,作淋巴细胞姊妹染色单体互换(SCE)和染色体畸变的检测。每份血分对照组(不加病毒悬液)和A、B、C实验组(根据加不同病毒量而分)。其结果:一、SCE频率,实验组A(8.9±0.19)、实验组B(9.9±0.2)、实验组C(11.6±0.22)与对照组(6.57±0.15)比较,A、B、C、实验组均分别高于对照组,P<0.01,差异有高度显著性,A、B、C三个实验组比较,P<0.01,差异有高度显著性。二、染色体畸变,A、B、C三个实验组分别与对照组比较,P>0.05,差异均无显著性。用EHFV HA 108株,ID_(50)10~(-6)/0.02ml接种2—5日龄大白鼠脑内,15天后颈动脉放血处死,取骨髓细胞培养,另取幼大白鼠骨髓细胞培养作对照,检测SCE和染色体畸变。结果:一、SCE频率,实验组(9.8±0.35)高于对照组(5.4±0.19),P<0.01,差异有高度显著性。二、染色体畸变,实验组与对照组比较,P>0.05,差异无显著性。以上两个实验结果表明,EHFV作用于细胞,无论是在机体或试管内,都引起SCE频率增高,即EHFV促使DNA产生初级损伤,但不致染色体畸变。     

Comparative Electrophoretic Studies of the Seed Proteins of Certain Species of Brassica and Sinapis

The saline-soluble seed proteins of Brassica campestris var.autumnalis, B. nigra, B. oleracea, B. campestris var. rapa andSinapis alba have been investigated using serological methodsand detection of enzymes after electrophoresis in starch gels.These methods were used to determine the generic status of Sinapisalba and the varietal or specific status of B. campestris var.rapa. The results of protein analysis agree with establishedtaxonomy and suggest that both methods help to resolve sometaxonomic problems.     

Comparative Electrophoretic Studies of the Seed Proteins of Certain Amphidiploid Species of Brassica

The saline-soluble seed protein extracts of two varieties ofBrassica nigra, B. Oleracea, B. campestris, B. carinata, B.juncea, and B. napus were investigated. The ß-galactosidases,ß-glucosidases, and esterases were detected afterelectrophoresis of the protein extracts in strach gels. Serologicalmethods were also applied to some of these extracts. The methodswere used in order to investigate the possibility of amphidiploidywithin the genus. The analysis of the seed proteins of suggestedhybrids with references to those of their putative parents isreported in this paper and gives evidence to support the suggestionof hybridization.