人血管瘤裸小鼠移植模型的超微结构变化

目的观察人血管瘤裸小鼠移植模型超微结构动态变化。方法将手术切取的增生期婴幼儿血管瘤组织移植到裸小鼠皮下,于移植后d2、d5、d15、d30、d45及第2、3、4、6个月切取移植瘤。戊二醛固定后行透射电镜观察。结果移植后早期内皮细胞大量变性、坏死。d30细胞出现增生表现,内皮细胞数量增加,部分细胞体积增大,核浆比增大,核质淡染。2个月时内皮细胞密集,部分内皮细胞变扁平。此后内皮细胞数量减少,出现胞质脱落、胞质固缩、次级溶酶体形成、染色体边集、核碎裂,伴基质中脂肪和胶原沉积。结论婴幼儿血管瘤组织移植到裸小鼠体内,经过缺血期后,瘤性内皮细胞可快速增生,继而缓慢消退,其超微结构变化与人血管瘤自然演变过程相似。     

环状RNA在小鼠肺发育中的连续表达及功能预测

目的 探讨环状RNA（circRNA）circ4:150439343|150477468和circ15:73330849|73343359在小鼠肺发育中的连续表达及潜在功能。方法 根据肺发育分期,取孕16.5 d（E16.5 d）、孕18.5 d（E18.5 d）及生后2 d（P2 d）胎鼠及新生小鼠的肺组织,利用苏木精-伊红染色观察肺组织形态学;利用qRT-PCR技术检测晚期肺发育过程中circ4:150439343|150477468和circ15:73330849|73343359的表达情况;应用miRanda和TargetScan软件预测circRNA的靶向miRNA,然后对靶基因进行GO和KEGG分析并预测相应circRNA功能。结果 在E16.5 d小鼠肺组织切片中观察到Ⅱ型肺泡上皮细胞,且逐渐增多,P2 d时肺泡迅速扩张,间质变薄,肺泡结构逐渐成熟;qRT-PCR结果显示circ4:150439343|150477468相对表达量随时间推移持续上调（P < 0.05）,circ15:73330849|73343359的相对表达量先下调再上调（P < 0.05）;宿主基因功能预测显示circRNA可参与Notch、PI3K-Akt和NF-κB等信号通路。结论 circ4:150439343|150477468和circ15:73330849|73343359可通过Notch等信号通路参与肺发育的调控。     

环状RNA在小鼠肺发育中的连续表达及功能预测

目的 探讨环状RNA（circRNA）circ4:150439343|150477468和circ15:73330849|73343359在小鼠肺发育中的连续表达及潜在功能。方法 根据肺发育分期,取孕16.5 d（E16.5 d）、孕18.5 d（E18.5 d）及生后2 d（P2 d）胎鼠及新生小鼠的肺组织,利用苏木精-伊红染色观察肺组织形态学;利用qRT-PCR技术检测晚期肺发育过程中circ4:150439343|150477468和circ15:73330849|73343359的表达情况;应用miRanda和TargetScan软件预测circRNA的靶向miRNA,然后对靶基因进行GO和KEGG分析并预测相应circRNA功能。结果 在E16.5 d小鼠肺组织切片中观察到Ⅱ型肺泡上皮细胞,且逐渐增多,P2 d时肺泡迅速扩张,间质变薄,肺泡结构逐渐成熟;qRT-PCR结果显示circ4:150439343|150477468相对表达量随时间推移持续上调（P < 0.05）,circ15:73330849|73343359的相对表达量先下调再上调（P < 0.05）;宿主基因功能预测显示circRNA可参与Notch、PI3K-Akt和NF-κB等信号通路。结论 circ4:150439343|150477468和circ15:73330849|73343359可通过Notch等信号通路参与肺发育的调控。     

体外血脑屏障模型的建立

目的：建立一种相对简单可靠、接近在体状态的SD大鼠体外血脑屏障模型。方法：原代分离、纯化培养大鼠脑微血管内皮细胞和星型胶质细胞,将两种细胞共培养建立体外血脑屏障模型。VIII因子、GFAP免疫组化鉴定细胞类型,相差显微镜下观察细胞形态,透射电镜观察内皮细胞超微结构及紧密连接,γ计数仪测定核素标记牛血清白蛋白(125I-BSA)通透量检测血脑屏障(BBB)功能。结果：VIII因子、GFAP免疫组织化学结果显示培养的脑微血管内皮细胞和星形胶质细胞纯度分别为90%和99%以上。透射电镜显示内皮细胞间有高度发达的紧密连接、未见有W eible-Palade小体、少见吞饮小泡1。25I-BSA通透量结果显示,血脑屏障模型组在限制大分子量物质的能力上优于其他各组,与脑微血管内皮细胞单独培养组及空白对照组比较,差异有显著性意义(P<0.01)。结论：利用同种属脑微血管内皮细胞与星型胶质细胞共培养,建立了一种简便可行、与在体血脑屏障结构类似,并具有在体血脑屏障的限制物质通透能力的SD大鼠体外血脑屏障模型。[中国当代儿科杂志,2005,7(6):526-529]     

卵清蛋白诱导宫内发育迟缓小鼠发生支气管哮喘动物模型的建立

目的 为了研究宫内发育迟缓（IUGR）与哮喘发生的相关分子机制,本研究在IUGR模型基础上建立卵清蛋白（OVA）诱导的支气管哮喘小鼠模型,并且对此模型进行了评价。方法 将受孕后的16只BALB/c雌鼠分成低蛋白饮食组和正常蛋白饮食组（n=8）。分别接受8%低蛋白饮食和20%正常蛋白饮食。仔鼠出生后6 h称体重。随机选取低蛋白饮食组中符合IUGR标准的16只雄性仔鼠纳入IUGR组,正常蛋白饮食组中16只雄性小鼠纳入对照组。留取两组小鼠血样测定血糖,ELISA法检测血清胰岛素水平。将对照组小鼠随机分为对照+PBS组和对照+OVA组（n=8）,将IUGR组小鼠随机分为IUGR+PBS组和IUGR+OVA组（n=8）。给予对照+OVA组和IUGR+OVA组6周龄小鼠腹腔注射2 mg/m LOVA致敏和雾化吸入1% OVA激发,对照+PBS组和IUGR+PBS组以等量PBS代替。ELISA法检测各组小鼠血清IgE水平;收集各组小鼠肺泡灌洗液行各类细胞计数;苏木素-伊红染色观察各组小鼠肺组织病理变化。结果 低蛋白饮食组仔鼠生后6 h体重低于正常蛋白饮食组（P < 0.01）。IUGR组小鼠血清胰岛素水平低于对照组（P < 0.01）。IUGR+PBS组IgE水平低于对照+PBS组（P < 0.01）;与对照+PBS组和IUGR+PBS组相比,对照+OVA组和IUGR+OVA组IgE水平均明显升高,且IUGR+OVA组IgE水平高于对照+OVA组（P < 0.01）。与对照+PBS组和IUGR+PBS组相比,对照+OVA组和IUGR+OVA组肺泡灌洗液中白细胞、嗜酸性粒细胞、淋巴细胞及巨噬细胞计数均明显升高（P < 0.01）。OVA诱导的IUGR小鼠肺泡组织呈现大量炎性细胞浸润,细胞间连续性被破坏;气道上皮细胞增生,支气管壁增厚,管腔狭窄;在支气管和血管壁周围亦观察到大量的炎性细胞浸润。结论 在低蛋白饮食建立的IUGR小鼠模型基础上,成功建立OVA诱导的支气管哮喘动物模型,为进一步研究IUGR和气道炎症之间的分子机制建立基础。     

22例小儿先天性心脏病右室超微结构的观察

近年来,小儿先天性心脏病的诊断和治疗取得了很大进步,但对心肌细胞超微结构形态改变研究不多,我们从1985年11月开始对22例先天性心脏病患儿进行心内膜、心肌活检,并经电子显微镜检     

胆红素、血脑屏障与核黄疸

血脑屏障由脑内毛细血管内皮细胞、基底膜和胶质膜组成。其生理意义在于保持中枢神经系统的恒定环境。血脑屏障发育不全或遭破坏时，低浓度的游离胆红素易进入脑组织，沉积于颅神经核，引起脑细胞能量代谢紊乱，严重者可产生核黄疸。了解其生理、生物特性对及时预报和防治核黄疸极为重要。     

促红细胞生成素通过早产儿血脑脊液屏障的观察

目的研究外源性重组人促红细胞生成素(rhu—EPO)能否通过早产儿血脑脊液屏障。方法将胎龄28-35周、体质量<2500g的早产儿随机分为治疗组20例,对照组16例。治疗组给予rhu-EPO750IU/(kg·周).3次/周,隔日1次,疗程2周;对照组按早产儿常规治疗。用酶联免疫法测两组早产儿治疗前后血清、脑脊液EPO浓度。结果1.治疗组治疗2周后血清、脑脊液EPO浓度均明显高于对照组(P均<0.01)。2.治疗组治疗前后血清、脑脊液EPO浓度相比,治疗后明显高于治疗前(P<0.01)。3.对照组血清、脑脊液EPO浓度在2周后同出生时相比均有所降低,但无显著差异(P>0.05)。结论外源性应用rhu—EPO能通过早产儿血脑脊液屏障。     

Abnormal Ultrastructural Features of Circulating Erythroblasts of the Laboratory Mouse with Hereditary Congenital Erythroblastic Anemia (hea/hea)

ABSTRACT Abnormalities of erythroid cells in the bone marrow of anemic CFO mice ( hea/hea ) with hereditary congenital erythroblastic anemia were not evident, while abundant erythroblasts, especially orthochromatophilic erythroblasts, were observed in the circulation and many showed abnormalities. Among them by electron microscopy, cytoplasmic aggregations of ribosomes and cytoplasmic lattice-like inclusions were characteristic of the hereditary congenital erythroblastic anemia.     

Pancreatic Cystadenoma in an Infant: Ultrastructural Study

A 10 × 6 × 4cm mullicystic cystadenoma arose in the pancreas of a 4-month-old male. Microscopically, the cysts were lined by short columnar or cuboidal cells that contained neither mucin nor glycogen. The lining cells had occasional nucleoli, contained electron dense vacuoles and apical aggregates of filaments, and were associated with a basal lamina. This is the youngest patient reported as having pancreatic cystadenoma, and the ultrastructure of the neoplasm was different from the 6 previously studied adult cases, suggesting that infantile and adult cystadenomas are different in nature.     

Ultrastructural Study of the Amniotic Epithelium in a Case of Gastroschisis

A distinct vacuolar change in the amniotic epithelium has been reported in association with gastroschisis. Ultrastructural examination of the placenta from a case with gastroschisis demonstrated numerous lipid droplets in the epithelial cells. The epithelial cells were otherwise intact and did not appear degenerated. The appearance of the lipid was different from meconium in macrophages in the underlying chorion.     

细菌性脑膜炎患儿血清、脑脊液基质金属蛋白酶水平变化及其临床意义

目的观察细菌性脑膜炎(PM)患儿血清和脑脊液(CSF)基质金属蛋白酶(MMP-2、MMP-9)水平的变化,分析其相关性,并结合CSF清蛋白指数(AQ),探讨MMP-2、MMP-9在PM患儿血脑脊液屏障(BBB)损伤中的作用机制,并分析其临床意义。方法采用ELISA法检测32例PM患儿(PM组)急性期、恢复期血清和CSF中MMP-2、MMP-9水平,采用溴甲酚绿法测定其血清清蛋白,免疫比浊法测定其CSF清蛋白。采用相同方法检测非神经系统疾病患儿(对照组,n=15)血清和脑脊液中MMP-2、MMP-9水平及清蛋白水平。结果PM组急性期AQ明显高于恢复期和对照组,差异均有统计学意义(Pa<0.001)。PM组急性期血清和CSF中MMP-2、MMP-9水平显著高于恢复期和对照组,差异均有统计学意义(Pa<0.001);PM组恢复期血清和CSF中MMP-2、MMP-9水平与对照组比较差异均无统计学意义(Pa>0.05)。PM组急性期、恢复期,血清MMP-2水平与CSF中MMP-2水平均无相关性(r=0.334、0.042,Pa>0.05);PM组急性期、恢复期,血清MMP-9水平与CSF中MMP-9水平呈正相关(r=0...     

Ultrastructural Criteria in Evaluating Leukemic Infiltration in Prepubertal Testicular Biopsies

The diagnosis of leukemic infiltration in the testis by routine histology is not always possible. During the past 5 years, 46 bilateral testicular biopsies from prepubertal boys were examined by electron microscopy for the purpose of establishing the presence or absence of leukemic infiltration. The ultrastructure of the cells of acute lymphoblastic leukemia is described from the positive cases¹⁰ and compared with cases from the literature, Variations in ultrastructure between the T-cell, B-cell, and “null cell” types are discussed. The ultrastructure of the normal prepubertal cellular elements, which include immature Leydig cells, primitive fibroblastic cells (intertubular), and attenuated peritubular fibroblasts, is described. Ultrastruclural criteria are summarized that enable a definitive evaluation of cases that are equivocal by histologic examination.     

B7同源体3经Toll样受体2依赖性机制增强肺炎链球菌脑膜炎小鼠的炎性反应和病理损伤

目的 探讨B7同源体3(B7 - H3)对肺炎链球菌(SP)脑膜炎大鼠的炎性反应和血脑屏障完整性的影响及其机制.方法 经小鼠侧脑室穿刺注入SP悬液,制备脑膜炎模型.野生型鼠分5组,分别为PBS组、SP组、SP+B7 -H3组、SP+同型单抗组、SP+B7 - H3阻断性单抗组.Toll样受体2基因剔除(TLR2-KO)鼠分3组:PBS组、SP组、SP+ B7 - H3蛋白组.术后6h、18h、30 h,麻醉其下眼眶采血法收集血清,取脑组织,制备脑组织匀浆.ELISA检测其血清TNF-α、IL-6、IL-1β和单核细胞趋化因子蛋白-1(MCP-1)水平以及脑组织匀浆中血清蛋白( Albumin)和IgG水平.结果 1.ELISA检测野生型鼠血清SP组、SP+ B7 - H3组注射18 h后TNF-α、IL-6和MCP-1的表达及30 h后TNF-α、IL-6、IL-1β和MCP-1的表达均较PBS组显著升高(Pa＜0.05);SP+同型单抗组与SP组比较,各时间点各炎性因子的表达差异均无统计学意义(Pa＞0.05);SP +B7- H3组注射18h后MCP-1的表达及30 h后TNF-α和IL-6表达均显著高于SP组[(42.010±3.883) ng·L-1vs(29.620±3.830) ng· L-1;(37.550±3.232)ng· L-1vs (24.570±2.377) ng·L-1;(66.160±5.766) ng·L-1vs (48.630±4.418) ng·L-1,Pa＜0.05];SP+ B7 - H3阻断性单抗组注射30h后,TNF-α和IL-6的表达均显著低于SP组[(18.680±1.798) ng·L-1vs(24.570±2.377) ng·L-1;( 37.180±3.150) ng·L- 1vs (48.630±4.418) ng· L-1,Pa＜0.05].2.ELISA检测脑组织匀浆:SP组、SP+ B7 - H3组注射18h、30h后Albumin、IgG的表达较PBS组均显著升高(Pa＜0.05);SP+同型单抗组与SP组比较,各时间点Albumin、IgG的表达差异均无统计学意义(Pa＞0.05);SP+ B7 - H3组注射18 h、30 h后脑组织匀浆Albumin的表达及30 h后脑组织匀浆IgG的表达均显著高于SP组[(59.090±4.184) μg· g-1vs ( 35.450±4.256) μg·g-1;( 59.890±4.701)μg·g-1 vs (43.790±3.508) μg·g-1;(36.220±2.775)μg·g-1 vs(25.440±2.620)μg·g-1,Pa＜0.05].3.SP+ B7 - H3阻断性单抗组注射后18 h、30 h Albumin的表达及30 h IgG的表达显著低于SP组[(20.590±1.720) μg·g-1vs(35.450±4.256) μg·g-1;(28.650±3.063) μg· g-1vs(43.790±3.508)μg·g-1;(17.380±1.595) μg·g-1vs(25.440±2.620) μg·g-1,Pa＜0.05].3.TLR2-KO鼠血清和脑组织匀浆的ELISA检测显示:SP +B7-H3组较SP组各监测指标的表达在任何时间点的差异均无统计学意义(Pa＞0.05).结论 B7 - H3通过TLR2依赖性机制促进SP诱导的脑膜炎小鼠的炎性反应和血脑屏障的破坏.     

小鼠胚胎心脏发育过程中胰岛素基因增强子结合蛋白1的时序表达与组蛋白乙酰化酶p300的关系

目的观察小鼠胚胎心脏发育过程中转录因子胰岛素基因增强子结合蛋白1(Islet-1)的时序性表达规律,并探讨其与组蛋白乙酰化酶p300介导的组蛋白乙酰化调控网络中的关系。方法以健康6～8周龄昆明小鼠为研究对象,下午1700雄雌按12比例合笼,次日观察阴栓,观察到阴栓最早之日计为胎龄0.5 d(E0.5)。取胎龄为E11.5、E14.5、E17.5的胎鼠和新生鼠的心脏,利用Western blot方法定量分析Islet-1的时序性表达规律,并利用蛋白质免疫共沉淀(Co-IP)和串联质谱分析方法(MS)鉴定Islet-1与组蛋白乙酰化酶p300的关系,同时应用Western blot方法反验证实验结果。结果 1.在小鼠胚胎心脏发育过程中,Islet-1在E14.5时的表达水平(0.434±0.353)明显高于与其在E11.5(0.074±0.456)、E17.5(0.120±0.127)和新生鼠(0.049±0.083)的表达水平,差异均有统计学意义(Pa<0.05),而其他时间点(E11.5、E17.5和新生鼠)间的表达差异无统计学意义(Pa>0.05)。2.在胚胎心脏发育过程中,Islet-1与组蛋白乙酰化酶p300相互结合,以蛋白复合物的形式存在。结论在小鼠胚胎心脏发育过程中,Islet-1可能作为枢纽因子,募集组蛋白乙酰化酶p300参与组蛋白乙酰化修饰调控。     

Developmental Analysis of Chlorambucil-Induced Occipital Blebs in Mice

SUMMARY This study was designed to examine whether defects of the meningeal anlage and intracranial hydrodynamic overload are involved in the pathogenesis of secondary cranioschisis after the neural tube closure. Chlorambucil (CA) 3.0–4.0 mg/kg administered to pregnant mice on day 7.5 of gestation (E7.5) caused abnormal blebs at the occipital region of embryos on E13.5 in a dose-dependent fashion compared with control. At blebs, the mesencephalic ventricle was dilated, particularly on the dorsal midline and posterior to the pineal invagination. Histology showed fewer mesenchymal cells and vessels, as well as thinner neuroepithelial tissue than in controls. Transmission electron microscopy revealed that the cytoplasm of mesenchymal cells in the bleb region contained abnormal electron-dense deposits, whereas deposits or other subcellular abnormalities were not evident in either the neuroepithelium or the surface ectoderm of the affected region. We administered 3.5 mg/kg of CA on E7.5 and studied the embryonic development. Mesodermal hypoplasia appeared on E11.5, whereas blebs were initially recognized on E13.5. On E15.5 and E18.5, the meninges and calvaria were hypoplastic, although no apparent cranioschisis was evident. To examine whether increased intracranial hydrodynamic pressure plus defects in the anlage of the meninges and cranium induces secondary cranioschisis, we injected kaolin into the cerebral ventricle on E13.5 to induce hydrocephaly. These embryos developed exo utero and we examined the morphology on E18.5. Hydrocephaly was induced and the defects were more severe in the meninges and cranium than in embryos exposed to CA alone, although cranioschisis was not histologically confirmed. These results suggested that hypoplasia in the mesoderm during early organogenesis underlies later abnormalities of the meninges and cranium, which may be involved in the pathogenesis of certain types of cranioschisis.     

Significance of X Granules in Histiocytosis X: An Ultrastructural Study

Histiocytosis X is characterized by the presence of cytoplasmic rod structures called Langerhans' cell granules or X granules (XG). It has been speculated that histiocytosis X is a Langerhans' cell disorder. This ultrastructural study was performed to quantitate the number of XG containing histiocytes in the histiocytosis X lesions. Twenty-four specimens from 22 patients with histiocytosis X were studied: 4 from skin, 5 from lymph node, 11 from bone, 2 from lung, 1 from gingiva, and 1 from cheek. The majority of the histiocytes in histiocytosis X lesions do not contain X granules. The percentage of histiocytes with XG in a lesion has no relation to the age of the patient or the organ from which it was obtained, except for skin, where they were quite numerous. The relative percentage of histiocytes with granules does not correlate significantly with the prognosis of these patients.