EL9611红白血病小鼠急性GVHD动物模型的建立

目的: 建立EL9611红白血病小鼠急性移植物抗宿主病(GVHD)的动物模型。方法: 同种异基因骨髓移植(allo-BMT)以C57BL/6(H-2^b)鼠为供鼠,BALB/c(H-2^d)为受鼠。设白血病组(n=10)、照射对照组(白血病鼠照射后不进行allo-BMT,n=4)、GVHD组(白血病鼠照射+allo-BMT,n=10)及正常对照组(n=4)。白血病组采用每只BALB/c鼠尾静脉输注2×10⁶个EL9611红白血病细胞建立红白血病动物模型;GVHD组于接种白血病细胞7 d后行总剂量为8.0 Gy的1次性 γ射线全身照射(TBI),照射后5 h内每只小鼠尾静脉输注C57BL/6鼠骨髓细胞2×10⁶个＋脾细胞1×10⁷个,建立EL9611红白血病小鼠的急性GVHD动物模型。观察小鼠体位、皮毛、大便等临床表现,病理检查肝脾、皮肤、小肠、外周血和骨髓,计算生存率。结果: 白血病组生存时间(14.5±2.1) d ,生存时间与GVHD组相比P<0.01,死亡率100％,无自发缓解,死亡时肝脾肿大(肝重2.40 g±0.48 g,脾重0.84 g±0.20 g,与正常对照组比P<0.01),外周血WBC升高 ,病理检查示组织正常结构破坏,白血病细胞浸润。照射对照组生存时间为(9.0±0.7) d,生存时间与GVHD组和正常对照组相比差异显著(P<0.01),死亡率100％,病理检查显示造血衰竭。GVHD组生存时间为(32.0±3.2)d,生存时间与其它各组相比P<0.01,死亡率100％,allo-BMT后第10-13 d出现症状,临床表现和病理检查符合Ⅰ到Ⅱ度GVHD的改变。结论: 采用EL9611红白血病细胞(2×10⁶/鼠)静脉输注的方式可成功建立EL9611红白血病动物模型;接种EL9611红白血病细胞第7 d行TBI +allo-BMT可成功建立EL9611红白血病小鼠的急性GVHD动物模型。     

BALB/c小鼠肥大细胞瘤/白血病模型的建立

目的研究建立BALB/c小鼠肥大细胞瘤/白血病模型。方法 BALB/c小鼠尾静脉注入小鼠肥大细胞瘤P815细胞,实验按每只小鼠接种细胞数量分为1×10^7/只组、5×10^6/只组、2.5×10^6/只组、1×10^6/只组、5×10^5/只组、1×105/只组和溶剂（RPMI1640培养液）对照组。观察小鼠的生存状态、体重及肝肺脾（重量、形态、病理）、染色体、血涂片、骨髓涂片、白细胞及血小板计数。结果注入P815细胞≥1×106/只的所有组小鼠均死亡,〈1×10^6/只的所有组生存良好,且死亡小鼠的生存时间与注入细胞数量呈负向依赖关系（P〈0.05）。死亡组小鼠体重较注射前相当或减轻、肝肺脾重较对照组及未死亡组显著增加（P〈0.05）;肝质地变硬,表面可见大量大小不一的白色结节突起,部分脾肺可见出血梗死灶,病理示肝脾有大量异形肥大细胞瘤细胞浸润;脾细胞染色体分析可见肥大细胞瘤细胞的非正常染色体核型;白细胞和血小板计数较对照组降低（P〈0.01）,血涂片可见少量肥大细胞瘤细胞,骨髓涂片示骨髓原始细胞比例增高。结论 P815细胞通过静脉注射能使BALB/c小鼠形成肥大细胞瘤/白血病,可作为肿瘤实验研究的一种模型构建方法。     

急性髓系白血病干细胞NOD/SCID小鼠白血病模型的建立

目的探讨用非肥胖型糖尿病/严重联合免疫缺陷（NOD/SCID）小鼠和经流式细胞仪分选出的KG1a中CD34＋CD38-的干细胞亚群建立白血病干细胞（LSCs）动物模型,为靶向治疗LSCs的药物筛选奠定体内实验基础。方法 18只雌性NOD/SCID小鼠,体质量18～20 g,鼠龄6～8周龄。实验组12只,应用流式细胞仪分选KG1a细胞中具有LSCs特性的CD34＋CD38-亚群,以2×106个/只尾静脉注射经全身X射线照射2Gy的NOD/SCID小鼠;正常对照组6只,只注射磷酸盐缓冲溶液。观察两组小鼠的一般情况和白血病发生情况,应用形态学、组织病理检查、流式细胞术、骨髓染色体检查等检测实验组小鼠的外周血、骨髓、肝脏、脾脏的白血病细胞标志。结果接种2周后实验组小鼠外周血可见白血病细胞,接种30 d实验组小鼠的白血病发病率为100%,无自发缓解;外周血、骨髓、肝、脾中均可发现大量的白血病细胞浸润;实验组小鼠骨髓细胞中CD13抗原阳性率15.47%～23.66%,并可见KG1a细胞的核型特征。结论尾静脉接种流式细胞仪分选后的CD34＋CD38-KG1a细胞于全身亚致死量X射线照射后的NOD/SCID小鼠,能成功建成全身播散的白血病模型,为进一步研究LSCs的靶向治疗药物奠定实验基础。     

虾类原肌球蛋白的Balb/c小鼠致敏动物模型构建研究

目的虾类主要过敏原原肌球蛋白(Tropomyosin,Tm)的Balb/c小鼠致敏动物模型的构建及Tm的低过敏性处理效果评价。方法本研究选用Balb/c小鼠,设立常见致敏蛋白卵清蛋白(OVA)组、虾主要过敏原Tm组、经无花果蛋白酶消减处理的Tm(Tm-E)和阴性对照PBS组,将各组样品与氢氧化铝佐剂(3∶1)混合后采用腹腔注射方式以构建Tm致敏动物模型,每次注射时间间隔为1周。第4天采集小鼠血清,通过ELISA评价血清中IgE和IgG抗体的效价变化。结果 OVA和Tm刺激38 d后产生大量IgE和IgG抗体;阴性对照组在整个刺激过程中其血清中不含有IgE和IgG抗体;Tm-E组在致敏24 d、38 d时其IgE的OD值分别较Tm组低77.83%和64.06%(P<0.01)。结论本研究成功构建了虾主要过敏原Tm的致敏动物模型,此外发现无花果蛋白酶解处理方式(酶活与底物比13.6 U/g,35℃酶解10 min)可有效降低Tm的过敏原性。     

杨树花粉过敏BALB / c 小鼠动物模型的建立

目的:建立杨树花粉粗提物致敏激发的小鼠过敏模型。方法:将60 只BALB/ c 小鼠随机均分为正常组、卵清蛋白(OVA)阳性对照组和模型组,每隔5 d 经腹腔注射致敏1 次,共4 次,末次致敏后第5 天滴鼻激发。观察支气管肺泡灌洗液(BALF)中炎症细胞变化情况;PAS 染色观察气道黏液分泌情况;HE 染色观察鼻黏膜、肺炎症情况;免疫组化法检测肺组织中IL-4 及IFN-α阳性细胞平均光密度值;酶联免疫吸附试验检测血清中的总IgE 水平。结果:与正常组比较,模型组诱导鼻黏膜、肺组织出现明显的变应性炎症且气道黏液分泌明显增加;BALF 涂片显示明显的炎症细胞(嗜酸性粒细胞、中性粒细胞)增多;肺组织中IL-4 阳性细胞平均光密度值均高于正常组,IFN-α阳性细胞平均光密度值均低于正常组;血清中总IgE 高于正常组。结论:杨树花粉粗提物能够成功建立小鼠过敏模型。该模型的建立有利于过敏性疾病机制的研究。     

Balb/c与DO11.10小鼠哮喘模型的建立

背景：哮喘的发病率逐年增高,建立和完善可以反映这种疾病的动物模型对于疾病研究具有重大意义。 目的：探讨Balb/c与DO11.10小鼠哮喘模型的诱导方法,比较两者的差异。 方法：24只Balb/c小鼠按随机数字表法等分为A-D组;用同样的方法把24只DO11.10小鼠等分为E-H组。A和E组小鼠第1天以PBS致敏,第8-11天以PBS雾化; B和F组小鼠第1天以卵清蛋白致敏,第8-11天以卵清蛋白雾化;C、G组小鼠第1,7,14天以PBS致敏,第22-25天以PBS雾化;D和H组小鼠第1,7,14天以卵清蛋白致敏,第22-25天以卵清蛋白雾化。 结果与结论：1次致敏加上4次雾化只能引起Balb/c小鼠轻度的炎症反应,在经历3次致敏和4次雾化后Balb/c小鼠出现了气道反应性增高,肺部炎症细胞浸润,支气管杯状上皮黏液分泌增加,肺泡灌洗液炎症细胞和嗜酸性粒细胞数量显著上升,同时肺泡灌洗液中Th2型细胞因子水平也升高。DO11.10小鼠经过1次致敏和4次雾化即出现了显著的炎症反应,值得注意的是,其肺泡灌洗液中性粒细胞的数量也显著上升;而在经过3次致敏和4次雾化后,DO11.10小鼠炎症反应反而明显减轻。提示卵清蛋白可成功诱导出Balb/c和DO11.10小鼠的哮喘模型,不过两者在诱导方案和病理表现上有所差异。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程      

Balb/c小鼠脾脏B10细胞分析

目的分析年龄、性别和环境因素对Balb/c小鼠脾脏B10细胞的影响。方法以年龄、性别和不同环境对小鼠进行分组,流式细胞仪分析小鼠脾脏CD5+CD1dhiB细胞的比例。体外培养小鼠脾细胞,在不加任何刺激或LPS+PIM刺激5 h后,流式细胞仪分析CD19+IL-10+B细胞比例。结果 6月龄小鼠脾脏中CD5+CD1dhiB细胞比例平均为2.9%,明显高于2月龄小鼠的2.28%(P<0.05);在不加任何刺激的情况下,两组小鼠脾脏中IL-10+B细胞的比例均很低且无统计学差异(P>0.05);加入LPS+PIM刺激5 h后,6月龄小鼠脾脏中IL-10+B细胞的比例平均为1.93%,高于2月龄小鼠的1.25%(P<0.05)。细胞表型和细胞内IL-10染色分析均显示年龄匹配的雌性和雄性小鼠脾脏B10细胞的表达无明显差异(P>0.05)。普通级和SPF级小鼠CD5+CD1dhiB细胞和LPS+PIM诱导的IL-10+B细胞比例无明显差异(P>0.05),而在不加任何刺激的情况下,普通级小鼠脾脏IL-10+B细胞比例平均为0.87%,明显高于SPF小鼠的0.38%(P<0.01)。结论脾脏B10细胞可随Balb/c小鼠年龄增大而扩增,B10细胞的表达与性别差异无明显关系,微生物环境可促进B10细胞的活化,但对B10细胞总数无明显影响。     

二甲基亚砜诱导小鼠红白血病细胞分化的差异蛋白质组学

目的利用二甲基亚砜(DMSO)诱导小鼠红白血病(MEL)细胞分化,系统地分析和鉴定诱导分化不同时间的差异蛋白质组学。方法 DMSO诱导MEL细胞分化,通过联苯胺染色、MTT比色法和Ter119免疫荧光等实验检测细胞分化的比率和细胞的活力,并利用双向电泳耦联质谱技术,结合生物信息学分析诱导MEL细胞分化过程中差异表达的蛋白质。结果 1.2%DMSO诱导MEL细胞分化,分别培养0h、6h、12h、24h、36h、48h、72h、96h、120h,利用双向电泳和质谱分析,共鉴定出87种蛋白质。1.2%DMSO作用MEL细胞120h后,细胞诱导分化率达到67%。MTT实验显示,1.0%、1.2%、1.4%的DMSO对MEL细胞的活力无明显的抑制作用。结论 DMSO对MEL细胞有诱导分化作用,但无明显地抑制细胞增殖。87种双向电泳差异蛋白质按功能可分为12类,比例最大的前3类分别是:占41%的酶类蛋白、15%的结构蛋白、13%的调节蛋白。     

脐血源性T淋巴细胞移植在BALB/c小鼠红白血病治疗中的作用

目的 研究脐血源性T淋巴细胞在小鼠体内的存活、浸润情况,以及对红白血病(EL)小鼠的治疗作用,为探讨脐血源性T淋巴细胞的生物学特性及其治疗白血病提供实验和理论依据.方法 48只BALB/c小鼠随机分为对照组、实验1组和实验2组.分离制备脐血源性T淋巴细胞,对其标记后经尾静脉或腹腔移植入红白血病BALB/c小鼠体内,采用免疫荧光、免疫组织化学、透射电镜等方法观察脐血源性T淋巴细胞在小鼠体内的存活时间、浸润状况及其对白血病小鼠脾脏结构的影响.结果脐血单个核细胞(CBMNCs)经植物血凝集素(PHA)诱导后,绝大多数细胞为CD3+ T淋巴细胞,阳性率可达(83.42±1.26)%;移植后第3、7天均可在小鼠外周血和肝、脾组织中检测到人CD3+细胞和Brdu+细胞;两实验组(EG)小鼠肝、脾组织内均见CD25+细胞散在浸润;两实验组EL小鼠平均存活时间分别为(27.25±7.06)d和(24.74±2.93)d,与对照组比较,均有统计学意义(P<0.05);Hoechst33258染色法检测显示,在荧光显微镜下见对照组细胞核荧光均匀,凋亡细胞较少;实验组部分白血病细胞呈现凋亡的形态学改变:染色质高度凝聚,致密浓染,或裂解成碎块状,边缘光滑清晰;透射电镜下见对照组小鼠脾内白血病细胞广泛浸润,实验组小鼠部分白血病细胞呈现典型凋亡形态学改变:核染色质浓缩、边集;核碎裂,可见凋亡小体,粗面内质网(RER)扩张,线粒体(Mi)呈现空泡样变及髓样变.结论脐血源性T淋巴细胞能在白血病小鼠体内存活并趋化至白血病细胞浸润部位,发挥其相应的功能,对白血病有一定的治疗作用.     

建立Alzheimer病动物模型的方法

随着人口老龄化的发展，老年性疾患已成为一个明显影响人类健康的突出问题，其中老年性痴呆（Seniledementia，SD）是最常见的老年性疾病之一，它和恶性肿瘤，心、脑血管意外并列为导致老年人死亡的三大疾病。据国外报道，SD患病率占老年人群4％～5％，其中半数以上为Alzh...     

Coxsackievirus B3-induced myocarditis: differences in the immune response of C57BL/6 and Balb/c mice

Coxsackievirus B3 (CVB3) infections are the most frequent causes of human myocarditis, often resulting in chronic stages characterized by fibrosis and loss of function. This disease is called dilated cardiomyopathy (DCM). Persistent virus in the myocardium may lead to chronic activation of fibroblasts, and subsequently, to fibrosis of the myocardium. Studies with immunodeficient mice have shown that certain defects of the immune system retard the rate at which virus is eliminated from the heart, thus leading to viral persistence. Therefore, we followed the immune response of two immunocompetent mouse strains (C57BL/6 and Balb/c) to CVB3 infection. These two strains have been reported to develop different immune responses to infections and we expected a similar reaction to viral infections as well. The two mouse strains recovered completely from CVB3 infection and expressed identical levels of cytokine mRNA in the heart. However, the virus in heart tissue decreased more slowly in Balb/c than in C57BL/6 mice. This was accompanied by a strong virus-specific IgG and weak IgM response in the C57BL/6 mice, in comparison to the Balb/c mice. We conclude, therefore, that viral-specific IgG is of importance for CVB3 elimination from infected hearts.     

大鼠生殖道炎症动物模型的构建

目的:构建苯酚胶浆致大鼠阴道及宫颈炎动物模型。方法:切除双侧大白鼠卵巢,注射苯甲酸雌二醇和氢化可的松,造成假动情期和全身免疫功能低下,然后每鼠阴道内注入苯酚胶浆0.1 ml,隔天一次共3次,每天观察并记录外阴变化,术后第15 d称重后眼球取血以显微镜计数法测定白细胞总数,断颈处死取阴道及宫颈进行病理学检查。结果:大鼠阴道口红肿、有的有脓性物流出;宫颈肥大,阴道及宫颈黏膜上皮细胞变性坏死或脱落,形成溃疡,膜下有充血水肿、炎细胞浸润、腔内有大量炎性或脓性渗出物及坏死组织;腺体有不同程度增生及鳞状化生;白细胞总数明显增高;体重减轻。结论:成功构建了阴道宫颈炎大鼠动物模型,为进一步研究HMGN家族等分子在调控女性生殖道炎症发生发展的机制研究打下了基础。     

Construction of protein profile classification model and screening of proteomic signature of acute leukemia

The French-American-British (FAB) and WHO classifications provide important guidelines for the diagnosis, treatment, and prognostic prediction of acute leukemia, but are incapable of accurately differentiating all subtypes, and not well correlated with the clinical outcomes. In this study, we performed the protein profiling of the bone marrow mononuclear cells from the patients with acute leukemia and the health volunteers (control) by surface enhanced laser desorption/ionization-time of flight mass spectrometry (SELDI_TOF_MS). The patients with acute leukemia were analyzed as unitary by the profiling that were grouped into acute promyelocytic leukemia (APL), acute myeloid leukemia-granulocytic (AML-Gran), acute myeloid leukemia-monocytic (AML-Mon) acute lymphocytic leukemia (ALL), and control. Based on 109 proteomic signatures, the classification models of acute leukemia were constructed to screen the predictors by the improvement of the proteomic signatures and to detect their expression characteristics. According to the improvement and the expression characteristics of the predictors, the proteomic signatures (M3829, M1593, M2121, M2536, M1016) characterized successively each group (CON, APL, AML-Gra, AML-Mon, ALL) were screened as target molecules for identification. Meanwhile, the proteomic-based class of determinant samples could be made by the classification models. The credibility of the proteomic-based classification passed the evaluation of Biomarker Patterns Software 5.0 (BPS 5.0) scoring and validated application in clinical practice. The results suggested that the proteomic signatures characterized by different blasts were potential for developing new treatment and monitoring approaches of leukemia blasts. Moreover, the classification model was potential in serving as new diagnose approach of leukemia.     

Development of atherosclerosis in Balb/c apolipoprotein E-deficient mice

BACKGROUND: Since its creation in 1992 by gene inactivation via gene targeting, the apolipoprotein E "knockout" mouse has become the most widely used rodent model for the study of atherosclerosis. Commercially available apolipoprotein E(-/-) mice are bred on a C57BL/6J background. The goal of the present study was to investigate the development of atherosclerosis in apolipoprotein E-deficient mice generated on a Balb/c background. METHODS: We compared serum cholesterol concentrations and the development of atherosclerotic lesions in heterozygous Balb/c [apolipoprotein E(+/-)] mice fed regular rodent chow, Balb/c apolipoprotein E-deficient mice fed regular chow, and Balb/c apolipoprotein E-deficient mice fed a high-fat diet for up to 30 weeks. Expression of the chemokine JE (murine homologue of MCP-1), as well as the adhesion molecules E-selectin, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1, in the aortas of knockout mice fed a high-fat diet was measured by enzyme-linked immunosorbent assay. RESULTS: Balb/c apolipoprotein E-deficient mice develop atherosclerotic lesions in a reproducible temporal and morphological pattern. Total serum cholesterol concentrations in Balb/c apolipoprotein E-deficient mice fed regular chow or a high-fat diet, respectively, closely parallel those reported for C57BL/6J apolipoprotein E-deficient mice. The expression of all three adhesion molecules in the aorta follows a similar temporal pattern, peaking in the first 15 weeks, whereas JE concentrations peak around 23 weeks. CONCLUSION: The availability of Balb/c apolipoprotein E-deficient mice will facilitate the study of atherosclerosis in a mouse strain that can concomitantly develop other pathological states that are not readily inducible in mice with the C57BL/6J background.     

Effect of starvation and refeeding on biochemical and immunological status of Balb/c mice: an experimental model of malnutrition

Context: Although new methods for the induction of malnutrition disorders in laboratory animals have been developed, the bulk of the models described in the literature are essentially based on dietary restriction/starvation principle. In this context, little data are available about the metabolic and the immune system parameters of Balb/c mice under starvation/refeeding.

Objective: This study examined the effects of starvation and refeeding on the biochemical and immunological status of undernourished Balb/c mice.

Methods: Female Balb/c mice, weighing 20?g, were starved for 3 days and then refed with commercial pelleted diet for 8 days. The variables considered were as follows: body weight; serum protein and amino acid concentrations; liver protein content, and cholinesterase and arginase activities; jejunal protein and DNA contents as well as oligosaccharidase levels; hematological parameters (bone marrow and peripheral blood cellularity); peritoneal macrophage activation; and humoral and cell-mediated immune functions.

Results: Profound alterations in both biochemical and immunological conditions appeared after the starvation period. Refeeding resulted in the normalization of serum albumin levels, the intestinal DNA content and the gut-mucosal associated enzymatic activities, the blood lymphocyte counts, and the number of peritoneal macrophages. The markers of liver metabolic function (cholinesterase and arginase activities), and those of bone marrow hemopoiesis and the adaptive immune response (T-dependent antibody titres and delayed-type hypersensitivity response) remained altered after refeeding compared with control mice.

Conclusion: These findings suggest that fasted mice can be used as an animal model of acute starvation that might prove useful in evaluating the effectiveness of nutritional and immunopharmacological interventions.     

Food proteins from different allergen families sensitize Balb/c mice to family-specific immune responses

Abstract

The classification of food allergens based on the structure and function of proteins contributes to the study of the relationship between bioinformatics and potential allergenicity of allergens. Food allergens always share sequence similarity with the allergens in the same allergen families. For that reason, food proteins from different allergen families may induce different patterns of immune responses in animal models. Female Balb/c mice (3–4-weeks-old) were sensitized with food proteins once per week for 4 weeks, and then challenged 2 weeks later (on Day 42 of study). Blood was collected (to obtain serum levels of histamine and protein-specific IgG₁ and IgE antibodies) and measures of vascular permeability were performed 20?min after the challenge. Five food proteins (11S globulin, OVA [ovalbumin], HAS [human serum albumin] and LRP [lysine-responsive storage protein] of different allergen families, and Cry 1Ab/Ac [crystal protein]) were used to assess patterns of immune responses for each allergen family and then bioinformatics and digestive stability in simulated gastric fluid were employed to assess the overall utility of the Balb/c. The assay results indicated that, in this model, histamine and protein-specific IgE antibody levels and vascular permeability could be used to identify allergenicity of 11S globulin, OVA, and PAP (potato acid phosphatase) only. However, the results of the protein-specific IgG₁ measures could only distinguish allergic food proteins with negative control. Based on bioinformatic analyses, the five different food proteins clearly induced distinct patterns of immune responses in the Balb/c model.     

炎症性肠病动物模型的研究概况

The etiology and pathogenesis of inflammatory bowel disease are up to now still not clear and definite. Establishing the ideal animal model to study its cause and pathogenesis of this disease is very important. The ideal animal model should have the same manifestation with human inflammatory bowel disease on clinical and pathologic feature etc. In this article, the method, the pathologic character isfics and concerning pathogenesis, of a few common useful experiment animal models are discussed.