血虚证的现代研究概述

很多学者从血虚证的发病原因、病理、动物模型制作及传统方药等方面在整体、细胞、分子等水平上对血虚证作了深入地研究,在血虚病理变化、动物模型制作及四物汤、当归补血汤等相关方药研究上取得了一定的共识。血虚时,机体造血功能抑制,免疫功能降低,红细胞膜的酶如Na~ 、K~ -ATP酶活性降低,水盐代谢紊乱及细胞内一些信使物质含量异常等。通过放血、化学损伤、放射线损伤等造成血虚证的动物实验研究,人们进一步发现了传统方药对血虚证的治疗作用机制。这些经典方药都能增强机体免疫,促进造血干祖细胞的增殖分化,促进血细胞的生成从而发挥补血作用。这些研究大大丰富和发展了传统的中医血虚理论,为疾病的预防、诊断、治疗、预后判断提供了更客观、更全面的指导。     

痛风性关节炎动物模型的改良

目的通过改良痛风性关节炎动物模型,制备出更符合人类痛风性关节炎机制的动物模型。方法通过Coderre法与次黄嘌呤相结合（模型B）、Coderre法与氧嗪酸相结合（模型A）的方法建立痛风性关节炎动物模型,采用全自动生化分析仪测定血尿酸水平,观察关节滑膜组织形态学改变及大鼠不同时相步态变化,并将两种方法加以比较。结果模型B组大鼠血尿酸水平显著降低,关节滑膜组织形态学及不同时相大鼠步态改变明显,与模型A组比较有统计学意义。结论Coderre法与次黄嘌呤相结合方法建立大鼠痛风性关节炎动物模型更符合人类痛风性关节炎机制的动物模型。     

血虚证的现代研究概述

很多学者从血虚证的发病原因、病理、动物模型制作及传统方药等方面在整体、细胞、分子等水平上对血虚证作了深入地研究,在血虚病理变化、动物模型制作及四物汤、当归补血汤等相关方药研究上取得了一定的共识.血虚时,机体造血功能抑制,免疫功能降低,红细胞膜的酶如Na^＋、K^＋-ATP酶活性降低,水盐代谢紊乱及细胞内一些信使物质含量异常等.通过放血、化学损伤、放射线损伤等造成血虚证的动物实验研究,人们进一步发现了传统方药对血虚证的治疗作用机制.这些经典方药都能增强机体免疫,促进造血干祖细胞的增殖分化,促进血细胞的生成从而发挥补血作用.这些研究大大丰富和发展了传统的中医血虚理论,为疾病的预防、诊断、治疗、预后判断提供了更客观、更全面的指导.     

慢性粒细胞白血病小鼠动物模型建立的研究进展*

慢性粒细胞白血病(chronic myeloid leukemia,CML)是造血干细胞(hematopoietic stem cells,HSC)恶性克隆性增殖引起的一种血液系统疾病。动物模型是研究CML发病机制及药物靶向治疗的重要载体和工具。研究表明,CML小鼠模型可以通过逆转录病毒介导、转基因和白血病细胞移植的方法建立。三种方法建立的CML小鼠模型均可用于CML发病机制及药物疗效评估研究。实验动物模型进一步通过血常规、血涂片和骨髓涂片、免疫学、分子生物学及病理学等检测手段,判断模型是否建立成功。本文就近年来CML小鼠模型的建立、鉴定及研究应用进展进行综述。     

肝旺痰阻型高血压大鼠模型的建立和评价研究

目的：建立和评价肝旺痰阻型高血压大鼠模型。方法：采用自发性高血压大鼠,以长期激怒联合高质饮食法建立肝旺痰阻的复合证候。通过观察大鼠性情动态的变化及体重、血压及血脂和血管紧张素II的变化,对高血压大鼠肝旺痰阻证型进行综合评价。结果：模型组大鼠在性情动态及体重、血压、血脂和血管紧张素II等方面均与对照组有较大差异（P〈0．05）,符合了中医肝旺痰阻证型的表现。结论：采用自发性高血压大鼠,以长期激怒联合高质饮食法,可建立肝旺痰阻型高血压大鼠动物模型。     

神经根型颈椎病动物模型研究进展

神经根型颈椎病(cervical spondylotic radiculopathy, CSR)是由颈椎小关节紊乱或颈椎间盘退行性改变而压迫神经根的一种疾病，典型症状表现为神经根支配区的颈肩部和上肢放射痛或麻木，严重影响着患者的生活质量。为深入研究该疾病病因病机、发展过程、健康防护和药物治疗等，正确地选择和建立符合疾病临床特征的动物模型是解决问题至关重要的环节之一。本文查询国内外相关文献报道，将CSR动物模型的制备归纳总结为单纯压迫模型(血管钳钳夹法、丝线结扎法、L形钢柱压迫法、硅胶片压迫法、自体骨压迫法、尼龙鱼线压迫法)、联合刺激模型(钳夹法联合铬制肠线压迫法、福尔马林定量滤纸刺激法、药物介导联合动静失衡法)和无创性干预模型(低头屈曲固定法)三类。将CSR动物模型制备进行总结概述，分析其特点和优劣势，以期为临床前研究选择适宜的动物模型和探索理想模型提供重要参考依据。     

优化胶粘贴法建立裸鼠胃癌原位种植模型

目的通过对两种胶粘贴法建立的胃癌原位种植动物模型的比较研究,为探讨胃癌的发病机制和实验治疗提供理想的动物模型。方法用OB胶和FS生物蛋白胶法分别建立胃癌原位种植动物模型,观察和比较两种方法所建立的模型肿瘤生长状况、转移情况和形态学变化。结果FS生物蛋白胶组未出现肿瘤大片坏死,腹水形成率为85.7%,幽门梗阻发生率为57.1%;而OB胶组肿瘤大片坏死发生率为100%,腹水形成率为14.3%,未出现幽门梗阻。FS生物蛋白胶组有三例出现了肺和脑转移。结论FS生物蛋白胶法建立的裸鼠胃癌原位种植动物模型能更好的模拟人胃癌患者的临床过程,为研究人胃癌转移机制和实验治疗提供理想的动物模型。     

四物汤对再生障碍性贫血小鼠骨髓细胞增殖的影响研究

目的研究四物汤对再生障碍性贫血（AA）小鼠骨髓细胞体外增殖的影响,探讨其治疗AA的机制。方法采用流式细胞仪、骨髓造血祖细胞培养等技术,检测四物汤对AA小鼠骨髓细胞的增殖变化。结果 四物汤能促进骨髓有核细胞进入G2／S期、增加CFU—GM、CFU-E、BFU-E集落数,且与自然恢复组有明显增强的差异。结论 四物汤在体内有促进AA小鼠骨髓细胞增殖的作用,为补血药治疗AA提供了实验与理论依据。     

结核病动物模型研究进展

结核病是由结核分枝杆菌感染引起的传染病,是危害人类健康的主要传染病之一。动物模型已经成为研究人类传染病的标准化工具。虽然对于结核分枝杆菌而言并没有真正意义的动物资源,但由于不同种类的动物,对分枝杆菌的敏感性不一样,因此可以成为结核病研究的有利工具。结核病最常用的实验动物模型包括小鼠、兔和豚鼠。每种动物有其自身特点,但并不能完全模拟人类疾病。通过建立结核病的动物模型,可以大大增加我们对疾病的病因、毒力和发病机制的理解。除了这三种模型外,非人灵长类也常被用于结核病的研究。本文总结了这几种结核病模型的研究状况。     

脾肾阳虚型溃疡性结肠炎大鼠模型的建立

目的建立脾肾阳虚型溃疡性结肠炎大鼠模型。方法通过灌服大黄水煎液、肌肉注射氢化可的松并结合TNBS(2,4,6-三硝基苯磺酸)混合乙醇灌肠建立脾肾阳虚型UC动物模型。将60只大鼠随机分为空白组、脾肾阳虚型UC模型7、14 d及21 d组,采用酶联免疫法检测各组大鼠血清中FT3、FT4、T的含量。结果与空白组比较,脾肾阳虚型UC模型7、14 d及21 d组大鼠血清中FT3、FT4、T的含量均有降低(P0.05);尤以模型21 d组差异显著。结论 FT3、FT4、T是脾肾阳虚的敏感指标,检测血清FT3、FT4、T可以更好地佐证脾肾阳虚;证明了脾肾阳虚型UC动物模型复建成功。     

Effect of Danggui Buxue Tang on immune-mediated aplastic anemia bone marrow proliferation mice

To investigate the pharmacological effects of Danggui Buxue Tang (DBT) on immune-mediated aplasia anemia mice. The model of immune-mediated aplasia anemia mice was induced by means of ⁶⁰Co γ-ray irradiation and mixed cells of thymus and lymphnode of DBA/2 mice infusion through tail vein, the parameters tested indices were as following: blood picture, bone marrow nucleated cell count (BMNC), murine colony-forming unit-megakaryocytes (CFU-GM) of bone marrow cells, murine colony-forming unit-erthroid (CFU-E) and burst forming unit-erythroid (BFU-E). The results showed that DBT could not only withstand significantly decreation of blood cells by immune-mediated, but also stimulate on the growth of bone marrow colony cell and increase the weight of hemopoietic progenitor of bone marrow. Therefore, DBT had an obvious treat effect on immune-mediated aplasia anemia models mice.     

Blast colony-forming cell binding from CML bone marrow, or blood, on stromal layers pretreated with G-CSF or SCF

Blast colony-forming cells (CFU-BL) represent a specific subpopulation of special primitive progenitors characterized by colony formation only in close contact with a preformed stromal layer. CFU-BL derived from bone marrow of chronic myeloid leukaemia (CML) patients have been proved to adhere poorly to bone marrow derived stromal layers suggesting that the appearance of progenitors and precursors in the circulation is due to a defective adhesion of these cells to the bone marrow microenvironment. In the present experiments the effect of short-term incubation of preformed normal bone marrow stroma on the adherence of CML derived CFU-BL was studied. For stroma cultures bone marrow cells were cultured in microplates in the presence of hydrocortisone. Cultures were used when stromal layers became confluent and no sign of haemopoiesis could be observed. CFU-BL were studied by panning plastic non-adherent mononuclear (PNAMNC) bone marrow or blood cells. 8.9 +/- 2.4 colonies/103 PNAMNC (six experiments) were formed from normal bone marrow on stromal layers and 4.8 +/- 2.1 colonies/103 PNAMNC (five experiments) from CML bone marrow. Colony formation from normal bone marrow was not increased if stromal layers were incubated with 100 ng/mL granulocyte colony-stimulating factor (G-CSF) or stem cell factor (SCF). Incubation of stroma with G-CSF or SCF, however, increased the colony formation of PNAMNC from CML bone marrow or blood significantly. These findings suggest that local concentration of haemopoietic growth factors at the time of panning may influence the attachment of CML progenitors to the stroma.     

Recognition of sickle cell anemia in skeletal remains of children

The present study discusses in detail the osteological changes associated with sickle cell anemia in children and their importance in differential diagnosis. Posterior calcaneal and specific articular surface disruptive metacarpal lesions are diagnostic for sickle cell anemia. Calvarial thickening, tibial and femoral cortical bone thickening, and bowing are of more limited utility in differential diagnosis. Granular osteoporosis, pelvic demineralization and rib broadening are nonspecific. Localized calvarial “ballooning,” previously not described, may have diagnostic significance. Bone marrow hyperplastic response (porotic hyperostosis) in sickle cell anemia produces minimal radiologic changes contrasted with that observed in thalassemia and blood loss/hemolytic phenomenon. Two other issues, the osteological criteria for discriminating among the anemias and the purported relationship between porotic hyperostosis and iron deficiency anemia, are also discussed. There is sufficient information to properly diagnose the four major groups of anemias, and further, to establish that iron deficiency is only indirectly associated with porotic hyperostosis. The hyperproliferative bone marrow response (manifest as porotic hyperostosis) to blood loss or hemolysis exhausts iron stores, resulting in secondary iron deficiency. Am J Phys Anthropol 104:213–226, 1997. © 1997 Wiley-Liss, Inc.     

Disruption of dihydronicotinamide riboside:quinone oxidoreductase 2 (NQO2) leads to myeloid hyperplasia of bone marrow and decreased sensitivity to menadione toxicity

Dihydronicotinamide riboside (NRH):quinone oxidoreductase 2 (NQO2) is a flavoenzyme that catalyzes the reductive metabolism of quinones. To examine the in vivo role of NQO2, NQO2-null (NQO2-/-) mice were generated using targeted gene disruption. Mice lacking NQO2 gene expression showed no detectable developmental abnormalities and were indistinguishable from wild-type (NQO2+/+) mice. However, NQO2-null mice exhibited myeloid hyperplasia of the bone marrow and increased neutrophils, basophils, eosinophils, and platelets in the peripheral blood. Decreased apoptosis of bone marrow cells and circulating granulocytes contributed to myeloid hyperplasia and hyperactivity of bone marrow in NQO2-null mice. The hematological changes in NQO2-/- mice were specifically associated with loss of the NQO2 gene because histological analysis of various tissues including spleen, thymus, blood cultures, and urine analysis demonstrated no sign of infection. NQO2-null mice also demonstrated decreased toxicity when exposed to menadione or menadione with NRH. These results establish a role for NQO2 in protection against myelogenous hyperplasia and in metabolic activation of menadione, leading to hepatic toxicity. The NQO2-null mice are a model for NQO2 deficiency in humans and can be used to determine the role of this enzyme in sensitivities to toxicity and carcinogenesis.     

Enhancement of gamma-glutamyl transpeptidase expression in bone marrow cells by colony stimulating factors

Studies on blood serum from mammary carcinoma (MC) hosts, which promoted gamma-glutamyl transpeptidase (GGT) expression by normal rat bone marrow cells in liquid culture, were extended to various granulocyte-macrophage colony stimulating factors (CSFs). GGT concentration per cell was found to increase (without change in total cell number) by incubation for 48 h with purified CSF-2 gamma and CSF-1 (but not interleukin-3), with human giant cell elaborated GM-CSF and L-cell conditioned medium, as well as with the 3 MC preparations (host serum, MC extract and MC conditioned medium). GGT-inducing ability (per milligram protein) ranked the 7 preparations in the same order as did their proliferative effect (number of colonies per milligram protein) in the standard mouse bone marrow agar culture system. The quantitative correlation between these two kinds of activities (linear for their logarithmic values) was highly significant, r = 0.976, p less than 0.001. The alkaline phosphatase concentration of bone marrow cells in liquid culture was also increased in the presence of the same 7 preparations, and this again was proportional (r = 0.985, p less than 0.001) to their colony stimulating potential.     

Cell transplantation in heart failure management

Heart failure is becoming a major issue for public health in western countries and the effect of currently available therapies is limited. Therefore cell transplantation was developed as an alternative strategy to improve cardiac structure and function. This review describes the multiple cell types and clinical trials considered for use in this indication. Most studies have been developed in models of post-ischemic heart failure. The transplantation of fetal or neonatal cardiomyocytes has proven to be functionally successful, but ethical as well as immunological and technical reasons make their clinical use limited. Recent reports, however, suggested that adult autologous cardiomyocytes could be prepared from stem cells present in various tissues (bone marrow, vessels, adult heart itself, adipose tissue). Alternatively, endothelial progenitors originating from bone marrow or peripheral blood could promote the neoangiogenesis within the scar tissue. Hematopietic stem cells prepared from bone marrow or peripheral blood have been proposed but their differentiation ability seems limited. Finally, the transplantation of skeletal muscle cells (myoblasts) in the infarcted area improved myocardial function, in correlation with the development of skeletal muscle tissue in various animal models. The latter results paved the way for the development of a first phase I clinical trial of myoblast transplantation in patients with severe post-ischemic heart failure. It required the scale-up of human cell production according to good manufacturing procedures, started in june 2000 in Paris and was terminated in november 2001, and was followed by several others. The results were encouraging and prompted the onset of a blinded, multicentric phase II clinical trial for skeletal muscle cells transplantation. Meanwhile, phase I clinical trials also evaluate the safeness and efficacy of various cell types originating from the bone marrow or the peripheral blood. However, potential side effects related to the biological properties of the cells or the delivery procedures are being reported. High quality clinical trials supported by strong pre-clinical data will help to evaluate the role of cell therapy as a potential treatment for heart failure.     

Regulation of eosinophilopoiesis in a murine model of asthma

Eosinophilic inflammation plays a key role in tissue damage that characterizes asthma. Eosinophils are produced in bone marrow and recent observations in both mice and humans suggest that allergen exposure results in increased output of eosinophils from hemopoietic tissue in individuals with asthma. However, specific mechanisms that alter eosinophilopoiesis in this disease are poorly understood. The current study used a well-characterized murine animal model of asthma to evaluate alterations of eosinophil and eosinophil progenitor cells (CFU-eo) in mice during initial sensitization to allergen and to determine whether observed changes in either cell population were regulated by T lymphocytes. Following the first intranasal installation of OVA, we observed sequential temporal elevation of eosinophils in bone marrow, blood, and lung. In immunocompetent BALB/c mice, elevation of bone marrow eosinophils was accompanied by transient depletion of CFU-eo in that tissue. CFU-eo rebounded to elevated numbers before returning to normal baseline values following intranasal OVA exposure. In T cell-deficient BALB/c nude (BALB/c(nu/nu)) mice, CFU-eo were markedly elevated following allergen sensitization, in the absence of bone marrow or peripheral blood eosinophilia. These data suggest that eosinophilia of asthma results from alterations in two distinct hemopoietic regulatory mechanisms. Elevation of eosinophil progenitor cells in the bone marrow is T cell independent and likely results from altered bone marrow stromal cell function. Differentiation of eosinophil progenitor cells and phenotypic eosinophilia is T cell dependent and does not occur in athymic nude mice exposed to intranasal allergen.     

Cytometry and Time-Dependent Variations in Peripheral Blood and Bone Marrow Cells: A Literature Review and Relevance to the Chronotherapy of Cancer

By flow cytometry of individual cells, multiple cell properties can be analyzed. Such parameters may be important in relation to cytotoxic treatment of cancer. For example, DNA measurements will answer questions regarding cell kinetics. Myelosuppression is the major dose-limiting toxicity during cancer treatment. Therefore, the study of cell cycle parameters in bone marrow cells is highly relevant. However, inattention to the existence and potential importance of biological rhythms may introduce artifacts and misleading results. The literature of rhythms in hematology is reviewed. Time-dependent variations in hematological variables have been extensively studied and rhythms have been described for all kinds of blood cells. Also the numbers of hemopoietic stem cells in the bone marrow undergo circadian variations. Our group has shown how such variations change with aging in mice. The relevance of time sequence studies in aging research of hemopoiesis was clearly demonstrated. In animal studies using cytometry, our group has demonstrated extensive circadian variations in cell cycle distribution of bone marrow cells, especially the DNA synthesis (S-phase). In humans a few and rather small time sequence studies of the bone marrow have been performed, so far. In this overview the clinical implications of circadian rhythms of S-phase variations measured by flow cytometry of human bone marrow cells are discussed. Male volunteers were examined every 4 h around-the-clock. The data indicated a lower proliferative activity during night, suggesting the possibility of reducing the bone marrow toxicity to cancer treatment when taking these time-dependent variations into consideration.     

Nutritional deficiency anemias in nonhuman primates

The purpose of this report is to review past studies in which anemias, occurred spontaneously in nonhuman primates due to feeding inadequate diets or were induced by feeding diets deficient in a nutrient. Included is a review of anemias induced by deficiencies of iron, niacin, pyridoxine, pantothenic acid, protein, riboflavin, cyanocobalamin, folic acid, ascorbic acid and alpha-tocopherol. The anemia induced by deficiency of each nutrient is discussed with emphasis on the major clinical signs as well as peripheral blood and bone marrow pathology. Results of supplementation of the diet following induction of deficiency states are discussed also. Whenever applicable, a discussion is included of the use of nonhuman primates as animal models for studies simulating parallel nutritional deficiencies in man.     

Transplantation with blood stem cells.

Peripheral blood stem cell transplantation (PBSCT) offers an alternative to autologous bone marrow transplants (A-BMT), especially in malignant diseases with bone marrow contamination. The presence of hemopoietic precursors in peripheral blood has been documented in several animal models and in humans. While many of these precursors might be committed cells with finite renewal capacity, ample evidence suggests that true pluripotent stem cells are circulating in a number sufficient to enable sustained trilineage engraftment after transplantation. Stem cell mobilization is markedly increased in the early recovery phase after intensive chemotherapy and can be promoted by the administration of various cytokines or polyanionic substances. These effects are used to optimize stem cell harvesting by leukapheresis. Clinical trials of PBSCT have been performed in several hundred patients with various hematological and nonhematological malignancies. Recovery was generally more rapid than after A-BMT. However, the envisioned advantage concerning disease control has not been documented so far.