地塞米松对脂多糖诱导的急性肺损伤幼鼠肺泡Ⅱ型上皮细胞超微结构的影响

肺泡Ⅱ型上皮细胞对维持肺表面活性物质的动态平衡和肺免疫功能具有极其重要的意义。地塞米松对急性肺损伤肺泡Ⅱ型上皮细胞超微结构的影响目前仍不清楚。该研究探讨急性肺损伤时及应用地塞米松干预后肺泡Ⅱ型上皮细胞超微结构的动态变化。方法：72只21 d SD幼鼠随机分为对照组、急性肺损伤组和地塞米松治疗组。肺损伤组的幼鼠腹腔注射脂多糖(LPS)（4 mg/kg）以建立急性肺损伤模型。对照组注射等量生理盐水。地塞米松治疗组在注射LPS 1 h后腹腔注射地塞米松（5 mg/kg）。每组随机选取8只幼鼠分别于注射LPS或生理盐水后24、48、72 h处死。取左肺下1 mm3的肺组织固定于2.5% 戊二醛中待透射电镜检查。结果：注射LPS 24 h后，肺损伤组大鼠AT II细胞微绒毛消失，板层小体数量增加。24 及48 h 板层小体呈指环状绕核排列。48 h出现巨大空泡变性样的板层小体。细胞核形态不规则，部分核边界不清。72 h 板层小体数目明显减少,核仁从细胞核消失,一些细胞核出现核溶解。注射LPS 后24 h地塞米松治疗组板层小体聚集在AT II细胞内同侧，并于该侧发生“胞吐”现象。48 h线粒体肿胀、聚集，脊断裂， 板层小体数目减少。72 h板层小体数目增多，并重新呈指环状绕核排列。结论：LPS诱导的急性肺损伤肺泡Ⅱ型上皮细胞超微结构的一系列变化呈时间依赖性。地塞米松能减轻急性肺损伤中肺泡Ⅱ型上皮细胞损伤和促进肺泡Ⅱ型上皮细胞的功能恢复。［中国当代儿科杂志，2007，9（6）：521-525］     

一氧化碳释放分子3通过抑制肺泡上皮细胞凋亡减轻脂多糖诱导的新生鼠急性肺损伤

目的 探讨一氧化碳释放分子3(CORM3)对脂多糖(LPS)诱导的新生鼠急性肺损伤(ALI)肺泡上皮细胞凋亡的影响。 方法 32只新生SD大鼠均分为对照组、LPS组、CORM3组和失活CORM3（iCORM3）组;LPS组、CORM3组和iCORM3组采用LPS气管内滴注建立新生鼠ALI模型,分别腹腔注射生理盐水、CORM3和iCORM3;对照组不建立ALI模型,腹腔注射生理盐水。于建模后12 h取肺组织,苏木素 伊红染色观察肺组织病理改变,湿干(W/D)比值测定,肺泡灌洗液(BALF)细胞计数及蛋白含量测定。体外培养A549细胞,LPS诱导细胞凋亡。MTT检测细胞活性,Tunel染色观察组织、细胞凋亡变化。 结果 ①与对照组相比,LPS组肺组织形态结构明显紊乱,肺泡压缩,肺间质大量炎性细胞浸润;CORM3组肺组织形态结构基本正常,间质炎性细胞浸润少;iCORM3组见肺组织水肿及大量炎性细胞浸润。②与对照组相比,LPS组肺组织W/D比值、BALF细胞数及蛋白含量明显升高,肺泡上皮细胞凋亡率明显增多［(37.3±4.5)% vs (3.0±1.0)%］;A549细胞活力下降,凋亡率明显升高［(29.6±4.1)% vs (3.6±1.0)%］,差异均有统计学意义 （P<0.05）。CORM3组肺组织W/D比值、BALF细胞数和蛋白含量较LPS组显著下降,肺泡上皮细胞凋亡率减少［（19.3±4.6）%］;A549细胞活力回升,凋亡率［（15.3±4.5）%］明显降低,差异均有统计学意义 （P<0.05）。LPS组与iCORM3组间上述指标差异均无统计学意义。 结论 CO通过抑制肺泡上皮细胞凋亡减轻新生鼠ALI     

支气管肺发育不良新生鼠模型肺泡上皮细胞标志物的表达变化及意义

目的 研究高氧致支气管肺发育不良(bronchopulmonary dysplasia,BPD)新生鼠模型中Ⅰ型肺泡上皮细胞(typeⅠalveolar epithelial cells,AECⅠ)和Ⅱ型肺泡上皮细胞(typeⅡalveolar epithelial cells,AECⅡ)特异性标志物表达变化及其意义.方法 80只新生Wistar大鼠,于生后12 h内随机分为模型组(吸入氧浓度为85%)和对照组(吸入空气),每组40只.于暴露第7、14、21天,分别进行肺组织HE染色观察病理学改变,免疫荧光双标染色观察AECⅠ标志物水通道蛋白5(aquaporin 5,AQP5)及AECⅡ标志物表面活性蛋白C(surfactant protein C,SP-C)表达及定位,Western blot方法检测AQP5和SP-C的蛋白表达水平,real-time PCR方法检测AQP5和SP-C的mRNA表达水平.结果 模型组大鼠肺组织表现出肺泡数目减少,体积增大,结构简单化,肺泡间隔增厚,次级分隔减少等肺泡化障碍表现.免疫荧光双标染色可见,模型组AQP5及SP-C表达明显增多,表达位置紊乱,且双染细胞/SP-C阳性细胞的比例明显增高(P<0.001).与对照组比较,模型组中AQP5及SP-C蛋白表达从高氧暴露7 d开始增加,增高的趋势持续至21 d.模型组中mRNA表达水平,AQP5从暴露7 d开始,SP-C从14 d开始较对照组明显升高(P<0.05),且两组间差异随高氧暴露时间延长更加明显(P<0.05).结论 暴露高氧中的新生鼠BPD模型肺组织中,AECⅠ标志物AQP5及AECⅡ标志物SP-C均表达上调,发生转分化的AECⅡ明显增多,表明在BPD肺损伤后的修复中存在AECⅡ过度转分化现象.     

急性肺损伤幼鼠肺泡Ⅱ型上皮细胞和SP-A变化相关性的研究

目的：该实验旨在研究急性肺损伤（ALI）时肺泡Ⅱ型上皮细胞（AEC-Ⅱ）超微结构变化和肺组织表面活性蛋白SP-A含量的变化关系，从而探讨ALI的发病机制。方法：48只Sprague-Dawley幼鼠被随机分为正常对照组和ALI组。 腹腔注射脂多糖（LPS，4 mg/kg）建立ALI模型，正常对照组注射等量生理盐水。 LPS注射后24，48，72 h每亚组各处死8只大鼠。 取左肺下肺组织待透射电镜检查。 用Western blot方法测定肺组织SP-A的相对含量。结果：ALI 24 h时，AEC-Ⅱ微绒毛消失。24 h及48 h时板层小体（lamellar body, Lb）数量增加，体积增大，密度减低，排空明显增强，呈指环状绕核排列，细胞增生活跃，代谢旺盛。48 h时Lb呈巨大空泡样变性。肺组织SP-A含量明显高于对照组（24 h时ALI组为6.52±0.62，对照组为5.02±0.35， P< 0.01；48 h时ALI组为6.65±0.62，对照组为5.01±0.36，P< 0.01）。72 h时Lb破溃，数目明显减少，细胞核形态不规则，部分核边界不清，肺组织SP-A含量下降(ALI组为3.87±0.50，对照组为5.22±0.36，P<0.01)。结论： LPS致幼鼠ALI时AEC-Ⅱ和肺组织SP-A的变化为时间依赖性，随AEC-Ⅱ损伤程度的加重肺组织SP-A由代偿转为失代偿，可能是发生ARDS的重要机制之一。     

高氧暴露对新生鼠Ⅱ型肺泡上皮细胞转分化水平的调节作用

目的 研究体内及体外高氧暴露对Ⅱ型肺泡上皮细胞(typeⅡalveolar epithelial cells,AEC Ⅱ)转分化水平的影响,旨在阐明支气管肺发育不良(bronchopulmonary dysplasia,BPD)肺上皮损伤的发生机制.方法 新生Wistar大鼠生后随机分为对照组(吸入空气)和模型组(吸入氧浓度为85％),于7d、14 d、21 d进行动物模型肺组织取材并分离AECⅡ.细胞标本检测Ⅰ型肺泡上皮细胞(type Ⅰalveolar epithelial cells,AEC Ⅰ)特异性标志物水通道蛋白5(aquaporin 5,AQP5)及AECⅡ标志物表面活性蛋白C(surfactant protein C,SP-C)表达.从正常新生鼠肺分离的AECⅡ在体外原代培养24h后随机分为常氧组(21％ O2)和高氧组(85％ O2),培养48 h后收集细胞,应用免疫荧光双标染色观察AQP5和SP-C表达及定位,Western blot方法检测AQP5和SP-C蛋白表达水平,荧光实时定量PCR方法检测AQP5和SP-CmRNA表达水平.结果 BPD模型组大鼠AECⅡ中AQP5蛋白表达从7d开始较对照组增多,SP-C蛋白表达从14 d开始较对照组减少.模型组中AQP5 mRNA从7d开始表达增多,SP-C mRNA从7d开始表达减少(P＜0.05),且随高氧暴露时间延长两组间差异更加显著.由正常新生鼠肺分离的AECⅡ进行体外原代培养后,免疫荧光双染可见高氧组较常氧组AQP5表达增多,SP-C表达减少,双染细胞明显增多.蛋白和mRNA定量检测结果均提示高氧组较常氧组AQP5表达增多,SP-C表达减少(P＜0.01).结论 无论体内还是体外高氧暴露下,AECⅡ特异性标志物SP-C表达下调,而AEC Ⅰ特异性标志物AQP-5表达上调,提示AECⅡ过度转分化参与高氧肺损伤后的修复过程.     

Notch受体在早产鼠高氧肺损伤中的动态表达

目的检测Notch受体、AQP5、SP-C在高氧暴露下早产SD大鼠肺的表达,探讨Notch在高氧肺损伤中的作用机制。方法SD早产鼠80只随机分为对照组和高氧组,于1、71、42、1 d行肺组织病理学检查;免疫组织化学法检测Notch1、Notch3;RT-PCR检测Notch1、Notch3、AQP5S、P-C mRNA;Western blot检测AQP5、SP-C。结果病理学检查显示,高氧组肺发育滞后,Ⅱ型上皮细胞(AECⅡ)分化抑制。免疫组织化学法检测结果显示,高氧组各时间点(除7 d)肺组织Notch1表达低于对照组(P<0.05,0.01),Notch3上调(P<0.01)。Notch1、Notch3 mRNA改变类似蛋白水平。高氧组AQP5,SP-C mRNA及蛋白较对照组显著下降。结论85%高氧导致早产鼠肺Notch受体表达异常。Notch信号可能通过调控转分化参与高氧肺损伤。     

高氧对早产大鼠肺表面活性蛋白C表达的影响

目的探讨高氧对早产大鼠肺表面活性蛋白C（SP-C）表达的影响。方法孕21dSD早产大鼠,生后12～24h内随机分为空气组、高氧组。于空气或高氧暴露后1、4、7、10和14d提取肺组织,采用RT-PCR测定SP-C mRNA表达,免疫组化和Western-blot检测SP＂C蛋白表达。结果早产大鼠生后肺组织SP-C表达1d时最高,4d后表达渐减弱,其阳性染色信号主要定位于Ⅱ型肺泡上皮细胞。高氧暴露1d,肺组织SP-C mRNA及蛋白表达显著低于空气组,以后增加,高氧7d增加最明显,高氧14d时SP-C表达较空气组又减弱。结论SP-C参与了早产大鼠肺发育及高氧肺损伤的生理与病理过程,高氧暴露导致SP-C表达下调或功能障碍是促使高氧肺损伤发生发展的重要因素。     

利多卡因对高氧暴露早产大鼠肺泡Ⅱ型上皮细胞损伤的保护作用

目的：观察利多卡因（Lido）对高氧暴露的早产大鼠肺泡Ⅱ型上皮细胞(AECⅡ)增殖和凋亡的影响,为防治早产儿高氧肺损伤提供依据。方法：原代培养的早产大鼠AECⅡ随机分为4组：空气组、高氧组、空气+Lido组、高氧+ Lido组。高氧、高氧+Lido组按5 L/min通入95%O2/5%CO2高纯混合气,10 min后密封。空气+Lido、高氧+Lido组加入20 μg/mL Lido。各组均置于培养箱(37℃,5%CO2)中培养24 h,收集各组细胞,采用流式细胞仪检测AECⅡ凋亡率和细胞周期;运用Western blot 检测增殖细胞核抗原（PCNA）蛋白表达。结果：与空气组比较,高氧组AECⅡ凋亡率增高,G2/M、S期细胞比例明显降低（P<0.01）;PCNA蛋白表达明显下调（P<0.01）。而Lido干预后可使AECⅡ凋亡率降低,G2/M、S期细胞比例增高,PCNA蛋白表达上调。结论：高氧可使早产大鼠AECⅡ凋亡增加,增殖抑制; Lido对高氧所致的AECⅡ损伤有直接的抑制作用。     

高氧诱导新生鼠肺上皮细胞凋亡损伤与肺表面活性蛋白的变化

目的探讨高浓度氧致新生鼠肺损伤时肺表面活性蛋白C、D(SP-C、SP-D)以及肺泡上皮细胞凋亡率的变化。方法生后24 h内的新生大鼠,随机分为空气组和高氧组;空气组常规饲养,高氧组置常压高氧箱内吸入浓度为90%的氧;两组分别于实验第1、3、7、10、14天,取8只新生鼠的肺组织进行苏木精-伊红(HE)染色并观察其病理变化,采用末端标记法检测肺上皮细胞凋亡率。采用酶联免疫法检测支气管肺泡灌洗液(BALF)中SP-C、SP-D。结果空气组新生鼠随日龄增加肺泡逐渐形成,形态规则,大小均匀;高氧组新生鼠随日龄增加肺泡数量逐渐减少,小血管扩张,出血增多,间质细胞增多,肺组织水肿。空气组新生大鼠BALF中的SP-C随日龄增长逐渐降低;高氧组第1天SP-C低于空气组,第3天SP-C高于空气组,第7天达高峰,第10天SP-C的含量开始下降,第14天下降更加明显。空气组SP-D的含量随日龄的增长亦逐渐降低;高氧组第1天SP-D的含量与空气组无明显差异,第3天SP-D的含量开始增加,第7天达到峰值,第10天SP-D的含量开始下降,第14天下降显著。结论长期吸入高浓度氧抑制肺泡发育,随吸入高浓度氧时间的延长,肺上皮细胞凋亡率增加,肺组织中SP-C、SP-D先增高后下降。     

高氧致新生鼠慢性肺疾病HoxB5 SPC AQP5表达及其意义

目的：肺泡上皮损伤是慢性肺疾病（CLD）主要病理改变之一,HoxB5基因是在肺发育过程中表达强烈的基因之一。该文探讨高浓度氧致新生鼠肺损伤时肺泡上皮损伤及修复能力,研究HoxB5在这一损伤中的动态表达规律及其与前者的关系。方法：建立高氧诱导新生鼠CLD模型, 80只早产鼠被随机分为实验组及对照组,分别应用免疫组化及RT-PCR技术测定肺组织HoxB5,SPC,AQP5蛋白及mRNA表达。结果：实验组肺组织HoxB5表达7 d后逐渐减少;高氧肺损伤SPC 3 d表达明显减少,7 d后开始增多,14 d、21 d明显高于对照组;而AQP5随着肺损伤加重表达量进行性下降。结论：高氧肺损伤新生鼠肺泡上皮细胞损伤明显,根据SPC,AQP5表达结果提示I型肺泡上皮细胞（AECI）损伤更甚且未得到正常修复,II型肺泡上皮细胞（AECII）虽然数量有所增加但其分化转化能力明显下降,而 HoxB5表达量减少可能致使大量的AECII失去了分化为AECI的功能。［中国当代儿科杂志,2009,11（1）：51-55］     

Stromal cell-derived factor-1α improves infarcted heart function through angiogenesis in mice

BACKGROUND: Local delivery of stromal cell-derived factor-1alpha (SDF-1) has been demonstrated to improve hind limb ischemia through enhanced neovascularization in animals. It was hypothesized that local administration of SDF-1 also contributes to neovascularization of ischemic heart. METHOD: Acute myocardial infarction was created by left coronary artery ligation in C57BL/6J mice. Immediately after infarction induction, mice were treated by injection directly into the center of ischemic myocardium either with saline (control group) or SDF-1 (SDF-1 group). Cardiac function was measured on echocardiogram 2 and 4 weeks after infarction. On week 4 mice were killed to evaluate infarction size and capillary vessel density. To determine the contribution of bone marrow cells to angiogenesis, the same procedures were performed on C57BL/6J chimeric mice reconstituted with green fluorescent protein-positive bone marrow cells. RESULTS: Fractional shortening was greater in the SDF-1 group at 4 weeks (0.31 +/- 0.06% vs 0.23 +/- 0.03%, P = 0.037). The infarct area was smaller in the SDF-1 group compared to the control group (9.31 +/- 2.76% vs 18.07 +/- 5.69%, P = 0.028). Green fluorescent protein-positive cells accumulated predominantly at the peri-infarction site, and were located with the capillary vessels. Capillary vessel density was significantly increased in the SDF-1 group (13.08 +/- 4.11 vessels/mm(2) vs 34.50 +/- 7.59 vessels/mm(2), P = 0.014). CONCLUSIONS: SDF-1 protects against deterioration of cardiac function after acute myocardial infarction by promoting angiogenesis. The safety and long-term prognosis of this treatment remains to be determined.     

ALA-PDT减轻同种异基因小鼠骨髓移植后移植物抗宿主病的实验研究

目的：探讨5-氨基乙酰丙酸-光动力疗法(5-aminolevulinicacid-mediatedphotodynamictherapy,ALA-PDT)在小鼠骨髓移植后减轻急性移植物抗宿主病(graft-versus-hostdisease,GVHD)的作用。方法：建立同种异基因小鼠骨髓移植急性GVHD模型,以C57BL/6J为供鼠,BALB/C为受鼠。BALB/C受鼠经致死剂量(8.5Gy)60Co照射后随机分为4组,A组为单纯60Co照射组;B组为单纯供鼠骨髓及脾细胞移植组;C组为供鼠骨髓移植以及供受鼠混合脾细胞移植组;D组为ALA-PDT移植组。在照射后4h移植供鼠骨髓细胞和脾细胞。观察骨髓移植后受鼠的一般情况、28d生存率、外周血白细胞计数以及肝脏、小肠、皮肤的病理组织学改变,应用流式细胞仪检测骨髓移植细胞嵌合情况。结果：D组小鼠28d生存率明显高于其他各组,差异有显著性(P<0.01),28d各组小鼠存活率分别为:0,0,10%,60%。B,C组小鼠均出现GVHD反应,其血液学以及病理组织学改变的严重度与GVHD发生的早晚具有一定的相关性。D组仅有2只小鼠出现GVHD反应,但其血液学以及病理组织学改变程度较其他各组轻。结论：ALA-PDT能够明显减轻小鼠骨髓移植后GVHD反应,增加受鼠的28d生存率,因此可能是一种有价值的抗同种异基因骨髓移植后急性GVHD反应的治疗方法。     

Cardiomyocyte regeneration from circulating bone marrow cells in mice

We investigated the role of circulating bone marrow cells (BMC) in cardiomyocyte regeneration. BMC, isolated from transgenic mice expressing enhanced green fluorescent protein (GFP), were transplanted into lethally irradiated C57BL6 mice. Five weeks after bone marrow transplantation (BMT), flow cytometric analysis for GFP-positive cells confirmed reconstitution of transplanted bone marrow. Bone marrow transplant mice subsequently underwent left coronary artery ligation (myocardial infarction) or sham-operation, and were killed at 1 mo or 3 mo after operation. Infarct size was similar in bone marrow transplant mice at 1 mo (47.1 +/- 5.9%) and at 3 mo (45.3 +/- 7.8%), and echocardiography at 2 and 8 wk revealed decreasing left ventricular function. In infarcted heart, GFP-positive cells that expressed desmin and troponin T-C were identified by confocal microscopy. GFP and troponin T-C double-positive cells were predominantly in the peri-infarcted region (1 mo, 365 +/- 45 cells/50 sections; 3 mo: 458 +/- 100 cells/50 sections; p < 0.05 versus noninfarct, infarct, and sham-operated regions). Furthermore, BMC mobilization and differentiation into cardiomyocytes was found to be complete within 1 mo after myocardial infarction. These results demonstrate that circulating BMC undergo mobilization and differentiation in cardiac cells after myocardial infarction. Future studies are required to determine the molecular signaling mechanisms responsible for this phenomenon.     

Recipient non-hematopoietic bone marrow cells in the intestinal graft after fetal small intestinal transplantation

We examined whether non-hematopoietic BM cells can migrate into the intestinal graft after fetal small intestinal transplantation (FSITx). Fetal small intestine from donor C57BL/6 mice was transplanted into the rectus abdominis of recipient C57BL/6 mice with only green fluorescent protein (GFP) BM cells (syngeneic FSITx). Intestinal grafts were harvested on days 5, 10, and 30 after FSITx and stained immunohistochemically using anti-CD45 antibody (a marker for hematopoietic BM cells). Although there were no GFP-positive cells identified in the epithelium of the graft intestinal villi, there were a few cells positive for both GFP and CD45 in the lamina propria on day 5 after FSITx, and many present on days 10 and 30. In some grafts there were only cells that were GFP positive/CD45 negative (i.e., non-hematopoietic BM cells) found in the lamina propria on days 10 and 30. These data indicate that non-hematopoietic BM cells as well as hematopoietic BM cells can migrate from the recipients bone marrow, suggesting that recipient mesenchymal stem cells may be strongly implicated in graft regeneration and development after FSITx.     

Comparison of Fatigue Resistant Properties of Gastrocnemius and Soleus Muscles between Muscular Dystrophic Mice and Other Strains of Mice

ABSTRACT: Muscular potentials were recorded from medial gastrocnemius (MG) and soleus muscles (SOL) of dystrophic and normal mice of the strain C57BL/6J- dy , dwarf and normal mice of the strain DW/J, and brachymorphic mice of the strain C57BL/6J-Zwi. The mice were anesthetized with urethane and the sciatic nerve was stimulated at 0.5 and 5 Hz with square pulses. When the frequency of stimulation was raised from 0.5 to 5 Hz and maintained at the latter frequency for 10 min, muscular potentials of dystrophic MG decreased slightly, whereas the other MGs showed a rapid and remarkable reduction in amplitudes. In addition, muscular potentials of SOL of all mice were slightly potentiated. Thus, changing patterns of muscular potentials of dystrophic MG were shifted toward those characteristic of SOLs.     

Ultrastructural studies on bone marrow development in embryonic mice.

The hematopoietic events in the embryonic C57B1/6J mouse bone marrow have been followed by electron microscopy. The first type of cells to appear in the embryonic bone marrow were mesenchymal cells identified at day 15 of gestation. The first recognisable hemopoietic cells to appear were those of the granulocytic series identified as mature cells at day 18, although single red cell precursors were also found at this gestational stage. The real erythroid development was noted on the last days of gestation (days 20 and 21), when also cells of the megakaryocytic series were found. At birth, day one of newborn life, all hematopoietic elements were present. The pattern of hematopoietic development in the embryonic bone marrow was compared to that observed in other embryonic hematopoietic sites.     

Cleft lip and palate in mice treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin: a morphological in vivo study

It is well-known that TCDD (2,3,7,8, tetrachloridedibenzo-p-dioxin) induces cleft palates (CPs) in pregnant C57BL mice. However, it is unclear if TCDD is a possible teratogen for cleft lip. We examined maxillofacial malformations including cleft lip in three animal strains: A/J mice, C57BL/6J mice and ICR mice. The A/J mouse develops cleft lip and palate spontaneously at a 5-10% rate. TCDD was administered in olive oil on gestation day (GD) 12.5 with gastric tubes at 10 microg/kg, 20 microg/kg, or 40 microg/kg to examine the dose-response, and on a single day from GD 8.5-14.5 to examine the timing effects of TCDD administration on lip and palate formation. Furthermore, the palatal shelf movements during GD 8.5-14.5 were observed with a stereoscopic microscope. All embryos had cleft palates when the TCDD was administered just before palatogenesis (GD11.5-GD12.5). With respect to the TCDD effects, there were large differences among the strains. In the A/J mice, the difference between a lethal dose and a dose that could induce a cleft palate was close. Cleft lips were not induced, even when the TCDD was given just before labiogenesis. Morphologically, both palatal shelves contacted perfectly along their lengths, but separated and formed cleft palates. In conclusion, TCDD is a strong inducer of cleft palates, and interferes with the fusion phase of the secondary palate, but has no effect on the lip.     

甲氧基聚乙二醇修饰对移植物抗白血病的作用

目的探讨甲氧基聚乙二醇(mPEG)修饰供者小鼠骨髓移植物单个核细胞表面抗原对移植物抗白血病作用(graft versusleukemiaeffect,GVL)的影响。方法小鼠随机分为A、B、C、D四组。A组单纯接受60Coγ照射;B、C、D组每只小鼠腹腔接种1×106L615肿瘤细胞制成白血病模型;全部照射后,C、D组分别移植mPEG未修饰和修饰的骨髓、脾细胞悬液。观察小鼠一般反应、外周血涂片L615细胞计数、组织病理及生存时间。结果A组全部死于造血衰竭;B组死于白血病;C组全部出现明显移植物抗宿主病(GVHD)表现并死亡;D组部分小鼠(4/15)长期存活,11/15死于白血病,平均生存24.2天,长于其他组(P<0.05),生存率27%,高于其他组(P<0.05),两者差异有统计学意义。结论小鼠移植mPEG修饰骨髓移植物后保留了一定GVL作用,并且减轻GVHD。     

白色念珠菌水溶物诱导小鼠冠状动脉损伤模型的建立

目的探讨建立白色念珠菌水溶物(Candida albicans water soluble fraction,CAWS)诱导幼年小鼠冠状动脉损伤模型的可行性。方法将48只4～6周龄C57BL/6小鼠随机分为实验组和对照组,各24只。先连续5 d分别给予实验组和对照组小鼠腹腔注射0.4 mg CAWS和磷酸盐缓冲液。完成最后一次注射后的24 h、3 d、7 d及28 d,两组各取6只小鼠处死,留取心脏标本,组织切片并苏木精-伊红(HE)染色;测量小鼠体质量、心脏质量及心脏/体质量比值。结果实验组小鼠3 d、7 d的体质量、心脏质量、及心脏/体质量比值较对照组降低,差异有统计学意义(P<0.05)。实验组在24 h、3 d时,6只小鼠中各有5只出现冠状动脉损害,表现为以中性粒细胞为主伴单核细胞浸润;7 d、28 d时,6只小鼠均出现冠状动脉损害,表现为近段扩张,炎症细胞以巨噬细胞为主,冠脉管壁出现成纤维细胞增生及弹力层黏液样变性等。实验组小鼠心脏病理变化类似川崎病冠状动脉损害。结论 CAWS连续腹腔注射能够诱导C57BL/6小鼠产生冠状动脉损伤的病理改变,建立一种表现与川崎病相似的动物模型。     

儿童白血病BMSCs及VLA-4抗体联合VP-16对白血病细胞凋亡的影响

目的研究儿童白血病(急性淋巴细胞白血病和急性髓系白血病)骨髓基质细胞(BMSCs)、缓慢抗原-4(VLA-4)抗体及VLA-4抗体联合化疗药物足叶乙甙(VP-16)对儿童白血病细胞凋亡的影响,探索儿童白血病新的治疗方法。方法分离培养儿童白血病骨髓单个核细胞,根据组别加入VLA-4抗体和VP-16,采用流式细胞仪检测不同组别白血病细胞的凋亡率。结果对照组与BMSCs组、骨髓基质上清组比较,其白血病细胞早期凋亡率均有显著增高,P<0.05,而BM-SCs组与骨髓基质上清组比较,白血病细胞早期凋亡率无显著差异。抗体组、BMSCs组和骨髓基质上清组白血病细胞凋亡率分别为[12 h:(10.16±1.29)%,24 h:(13.72±1.72)%]、[12 h:(4.99±1.66)%,24 h:(5.54±1.53)%][12 h:(6.06±1.77)%,24 h:(6.62±1.8)%],抗体组与BM-SCs组和骨髓基质上清组比较,其早期凋亡率均显著增加,P<0.05;抗体加药物组白血病细胞早期凋亡率最高,为[12 h:(26.79±1.29)%,24 h:(20.81±1.39)%],与其他任何一组比较,差异显著,P<0.05;除BMSCs组和骨髓基质上清组,其他四个组12 h与24 h的白血病细胞早期凋亡率有显著差异,P<0.05。结论白血病患儿的BMSCs可以保护白血病细胞免于失巢凋亡和药物诱导的凋亡,VLA-4抗体可增强VP-16诱导的白血病细胞的凋亡敏感性。