蛙心包淋巴孔的发现

首次报道蛙心包淋巴孔 ,揭示心包腔淋巴转归途径。本实验应用扫描电镜和透射电镜观察心包淋巴孔的超微结构 ,并使用计算机图象处理技术对淋巴孔作定量分析。结果发现 ,正常蛙心包腔面有一些散在分布的心包淋巴孔和少量淋巴窦。构成淋巴孔的间皮细胞常出现粗大的胞质突起 ,伸入淋巴孔 ,形成瓣膜状结构。淋巴孔的平均直径为 0 72±0 33μm ,平均分布密度是 3 57± 2 0 7个 /0 0 1mm2 ;心包间皮淋巴窦的面积是 995 0 8±2 2 1 74μm2 /0 0 1mm2 。蛙心肌无血管 ,其血供仅由心腔内血液直接进入心肌的小梁间隙。心包脏层未发现有淋巴孔。结果表明 :间皮淋巴窦是心包膜正常“漏出”的形态依据。心包淋巴孔的发现 ,证明心包腔淋巴引流途径的存在。淋巴引流对于心肌组织间液的平衡 ,清除组织间液蛋白质 ,防止心肌间质水肿 ,有重要意义     

小鼠膈腹膜淋巴孔和膈淋巴管的发育

应用透射电镜、扫描电镜和酶组织化学方法,研究胚胎期和出生后不同时期小鼠膈腹膜淋巴孔（PLS）和膈淋巴管的发生和发育,并用Elescope计算机图像处理技术对PLS进行定量分析。结果发现：胚胎13天时,膈腹膜仅由扁平形间皮细胞（FMC）组成;胚胎15天时,FMC间出现立方形间皮细胞（CMC）和早期腹膜淋巴孔（NLS）;胚胎18天时,膈毛细淋巴管出现,台盼蓝吸收试验显示NLS无物质吸收功能;出生后1天（PND1）,膈毛细淋巴管内皮细胞向PLS伸出胞质突起,并横跨CMC下结缔组织纤维和基底膜,形成腹膜下小管。后者与PLS沟通,建立了腹膜腔内物质转归通路。台盼蓝吸收试验表明,出生后PLS具有物质吸收功能,即为成熟腹膜淋巴孔（MLS）。PND5,立方细胞嵴（CMCR）发生,膈毛细淋巴管数量增多。PND10,大量立方细胞嵴融合,形成条带状分布的立方细胞区域,其上分布有大量MLS。随着发育进展,MLS平均面积和平均分布密度逐渐增大,且随着膈淋巴管的发育,吸收功能逐渐增强。     

一氧化氮对大鼠胸膜淋巴孔调控及淋巴吸收的影响

实验研究了一氧化氮（nitric oxide,NO）对大鼠胸膜淋巴孔的调控和胸膜腔淋巴吸收的影响。NO供体和NOS（nitric oxide synthase）抑制剂分别经腹腔给药,示踪剂（台盼蓝）胸膜腔内注射后,处死大鼠,测定血清NO和台盼蓝浓度;在扫描电镜下观察各组胸膜淋巴孔,用计算机图像处理,统计学分析。结果显示,NO供体组血清NO浓度为49.34±18.47μmol/L,淋巴孔的面积和密度分别为6.80±1.13 μm2和170.24±66.60/0.1mm2;NOS抑制剂组血清NO浓度为17.72±6.58μmol/L,淋巴孔的面积和密度分别为5.72±1.54μm2和61.71±12.73/0.1mm2。血清NO浓度与淋巴孔开放的面积和密度成正相关（P<0.05）。在胸膜腔给示踪剂后,NO供体组血清台盼蓝的浓度为74.68±33.67mg/L,与对照组比较有显著差异（P<0.05）。提示,NO可以调控胸膜淋巴孔,促进胸膜腔淋巴吸收。     

妊娠和激素干预对豚鼠卵巢囊淋巴孔的影响

本实验以台盼蓝为示踪剂,观察成年雌豚鼠在妊娠、注射促排卵激素和雄激素时,卵巢囊淋巴孔对示踪剂的吸收情况,并结合扫描电镜和计算机图像处理,对豚鼠卵巢囊内壁的淋巴孔进行观察和统计,以研究妊娠和激素干预对豚鼠卵巢囊淋巴孔开放和淋巴吸收功能的影响。结果表明,妊娠组卵巢囊吸收示踪剂的量最多,促排卵组次之,雄激素组最少,且组间两两比较均有显著性意义(P<0.05)。在每1000μm~2的视野中,妊娠组卵巢囊淋巴孔的开放面积为189.9±48.7μm~2,促排卵组为104.4±31.2μm~2,雄激素组为40.5±18.7μm~2,组间两两比较均有显著性意义(P<0.05)。妊娠组卵巢囊内层纤毛柱状上皮的分泌颗粒数量最少,而促排卵组的分泌颗粒较多,且纤毛最活跃。本实验研究结果表明,妊娠和激素干预能影响豚鼠卵巢囊淋巴孔的开放和吸收,囊内卵巢的功能状态与豚鼠卵巢囊淋巴孔的调控密切相关。     

腹膜淋巴孔与淋巴转归NO-cGMP-Ca2+信号转导途径研究

为研究一氧化氮(nitric oxide,NO)调节大鼠腹膜淋巴孔和淋巴吸收的细胞内信号转导机制,在体外培养的间皮细胞上,利用cGMP检测试剂盒和激光共聚焦扫描显微镜,研究NO对间皮细胞内cGMP和游离钙离子浓度(Ca^2 )的影响;并利用动物实验研究NO-cGMP-Ca^2 通路对大鼠腹膜淋巴孔和淋巴吸收的影响。结果发现：(1)与对照组相比,NO供体Sper NO （spermine／nitric oxide complex)可以剂量依赖性地升高间皮细胞内cGMP的浓度(P＜0．01)。此作用可被可溶性鸟苷酸环化酶(soluble guanylyl cyclase,sGC)特异性抑制剂1H-[1,2,4]噁二唑[4．3-a]喹喔啉-1-one酮(1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one,ODQ)阻断(P＜0．05,P＜0．01)。Sper／NO可使间皮细胞内[Ca^2 ],相对水平降低(P＜0．05),但此效应可被ODQ阻断;L-型电压依赖性钙通道阻滞剂nifedipine,可使细胞内的[Ca^2 ];在短时间内迅速明显下降(P＜0．05),达平衡后再加入Sper／NO并不能引起[Ca^2 ];的进一步改变(P＞0．05);(2)NO可以剂量依赖性地提高大鼠膈组织淋巴孔最大分布面积(P＜0．01)和对示踪剂的吸收量(P＜0．05),ODQ可明显抑制NO引起的淋巴孔和淋巴吸收的改变(P＜0．01)。该结果首次发现nifedipine能显著增加淋巴孔最大分布面积(P＜0．01)及膈组织对台盼蓝的吸收量(P＜O．05),而且nifedipine预处理后Sper／NO并不能使淋巴孔的淋巴吸收进一步升高(P＞0．05)。结果提示,NO可以通过降低cGMP水平降低大鼠腹膜间皮[Ca^2 ],且此过程和L-型电压依赖性钙通道有关;NO可通过NO-cGMP-[Ca^2 ],通路,促进淋巴孔的开放和吸收。     

家兔生后外周淋巴器官的组织形态学研究

本文探讨了不同发育阶段家兔淋巴结和脾的组织学年龄性变化,研究结果表明:1.初生时期淋巴结和脾脏均未达到成年水平,淋巴结皮质较薄,且呈弥散性;髓质琉松,含结缔组织较多,淋巴细胞稀少。2.具有生发中心的淋巴小结和脾小体在30日龄时出现。3.网状纤维在淋巴结主要分布于淋巴小结的外周;在脾白髓区域的网状纤维丰富而粗大。     

链脲菌素诱发新生期大鼠糖尿病的量效关系及其胸腺淋巴细胞增殖活性的变化

目的　研究链脲菌素 (STZ)处理新生期大鼠后诱发成年发病糖尿病的量效关系及其胸腺淋巴细胞增殖活性的变化。方法　STZ分别以 2 0、40、6 0及 10 0mg kg ,ip ,给予Wistar新生期大鼠 ,于注射STZ后第 3天、第 7天、第 17天、第 42天观察体重、血糖、血浆胰岛素及胸腺淋巴细胞增殖反应的动态变化。结果　 10 0mg kg剂量组可于第 42天形成慢性糖尿病模型 ,血糖值 (16 8± 4 6 )mmol Lvs (5 8± 1 6 )mmol L ,P <0 0 0 1;血浆胰岛素水平(5 2± 1 2 )mIU Lvs(8 4± 1 6 )mIU L ,P <0 0 1) ,胸腺淋巴细胞增殖呈现升高、受抑制再恢复的变化过程。其他各剂量组 ,特别是 40mg kg虽无明显高血糖形成 ,但可使胸腺淋巴细胞增殖活性增强。结论　STZ(10 0mg kg,ip)处理新生期大鼠诱发糖尿病的最适剂量为 10 0mg kg ,诱发时间为 42d。诱发过程中伴有胸腺增殖活性的变化。     

环境因子对大石鸡种群遗传结构的影响

大石鸡是我国特有种 ,仅分布于青海东部 ,甘肃中部 ,宁夏六盘山以西 ,是我国北方干旱和半干旱荒漠环境指示鸟类。研究其遗传多样性与环境变化的关系 ,不仅是生态遗传学的前沿领域 ,而且在进化生物学和保护生物学领域都有重要的理论意义。采用聚合酶链式反应 ( PCR)和直接测序的方法 ,测定了甘肃境内由北向南的 5个大石鸡 ( Alectorismagna)种群 (兰州、榆中、定西、武山和礼县 )的线粒体 DNA( mt DNA)控制区 ( D-loop) 4 5 7～ 4 5 8个碱基长度的基因序列。结果表明兰州、榆中、定西、武山和礼县种群的平均碱基含量中 A( F=0 .30 F0 .0 1( 4 ,32 ) =3.97)、T( F=6 .4 4>F0 .0 1( 4 ,32 ) =3.97)差异极显著。 5个种群的基因变异率分别为 0 .32± 0 .2 7%、0 .4 8± 0 .4 5 %、0 .6 2± 0 .4 3%、0 .4 4± 0 .2 4 %、0 .1 7± 0 .1 4 % ,种群内的平均基因变异率为 0 .4 1± 0 .1 7% ,种群间的平均基因变异率为 0 .4 6± 0 .1 0 % ,种群内和种群间的平均基因变异率差异不明显( F=2 .5 90 .0 5 )和无霜期 ( r=-0 .81 0     

胃癌nm23H1 mRNA、VEGF-C mRNA表达及其与淋巴转移、生存率的相关性研究

研究胃癌组织nm23H1 mRNA、VEGF-C mRNA和VEGFR-3的表达与胃癌中的新生淋巴管的生成、肿瘤淋巴道转移和生存率关系,为临床治疗及预后提供实验依据。用胃癌与癌周组织作对照,应用原位杂交法检测78例胃癌组织nm23H1 mRNA、VEGF-C mRNA表达,并以VEGFR-3抗体为标记,经EnVisionTM免疫组织化学及Leica-Qwin计算机图像分析,用Weidner最高血管密度计数法,计数阳性淋巴管数(MLC),以及调查患者术后的生存率。胃癌nm23H1 mRNA阳性表达 69．23％(54例)、nm23H1 mRNA阳性表达与胃癌淋巴结转移、TNM分期、MLC呈负相关(P<0．01), VEGF-C mRNA阳性表达46．15％(36例),其与胃癌淋巴结转移、TNM分期、MLC呈正相关(P<0．01 或P<0．05)．与癌周组织的nm23H1 mRNA和VEGF-C mRNA表达比较,具有显著性差异(P<0．01 或P<0．05)。nm23H1 mRNA在Ⅰ、Ⅱ期胃癌中呈高表达,在Ⅲ、Ⅳ期患者中呈低或无表达。胃癌组织的MLC(8．37±2．29/mm2)显著高于癌周组织(4．51±2．64/mm2),两者比较具有显著性差异(P<0．05)。 nm23H1 mRNA与VEGFR-3表达之间存在负相关(P<0．05,r=0．8479);VEGF-C mRNA与VEG- FR-3表达之间存在正相关(P<0．05,r=0．8362)。在根治术5年内死亡的61．54％(48例)患者中 MLC(10.82±2．51/mm2)高于5年内生存的患者(6．53±2．09/mm2),两组间有显著性差异(P<0．05);在不同的肿瘤病理学分级中,胃癌未、低分化型与高、中分化型的nm23H1 mRNA阳性表达,有显著性差异(P<0．05),当nm23H1 mRNA高表达时,肿瘤淋巴转移率低,生存率高,故认为nm23H1基因具有抑制胃癌发生和淋巴道转移作用;当VEGF-C mRNA高表达时,肿瘤淋巴转移率高,生存率低。胃癌组织中的VEGFR-3表达水平与肿瘤的淋巴转移密切相关,胃癌组织MLC的显著增高,提示肿瘤组织有新淋巴管的生成。VEGF-C促进了肿瘤诱导的淋巴管新生,在胃癌的淋巴道转移中起重要作用。     

四十里湾栉孔扇贝清滤率、摄食率和吸收效率的现场研究

运用生物沉积法在四十里湾不同海区对栉孔扇贝的生理生态学特征进行了研究。 1龄栉孔扇贝 (4 1 .1± 4.1 mm,软体干重 0 .48± 0 .1 0 g/ ind)清滤率变化范围为 0 .72～ 2 .5 4(平均 1 .2 7) L/ (ind· h)或 1 .65～ 5 .97(平均 2 .61 ) L/ (g· h)。清滤率受 TPM的变化影响不大 ,而摄食率却随 TPM的升高而升高。 2龄扇贝 (软体干重 1 .91± 0 .3 2 g/ ind)清滤率为2 .0 9～ 3 .99(平均 3 .1 0 ) L/ (ind· h)。栉孔扇贝吸收速率与 POM呈正相关关系 ,而与饵料质量 (POM/ TPM)无明显的相关性。1龄扇贝和 2龄扇贝吸收效率没有显著差别 ;扇贝对 POM的吸收效率与 TPM (或 POM)关系不大 ,却与饵料质量呈明显的正相关关系 ;扇贝对 POC、PON和 PP的吸收效率平均分别为 68.9%、64.0 %和 63 .6%。在沿岸养殖海域 ,栉孔扇贝通过大量的滤水摄食以及较高的吸收效率对生态系统的能量流动和物质循环产生影响。     

The ultrastructure of lymphatic valves in the adult rabbit lung

Summary An electron microscopic investigation has revealed that the pulmonary lymphatic valves of adult rabbits are not simple duplicatures of the lymphatic vessel wall. They consist of an uninterrupted central connective tissue core, covered on both sides with a single layer of flattened endothelial cells. Near their insertion in the lymphatic vessel wall, the connective tissue core reveals a distinct thickening being composed mainly of collagen bundles. In the other parts it contains mainly elastic fibers and fine filaments, enclosing also some rather peculiar connective tissue cells. Nervous and muscular elements were not observed. The endothelium is continuous and exhibits no open junctions. The valvular basement membrane is better developed than in lymphatic capillaries. The endothelial cells contain numerous cytoplasmic filaments which might be endowed with contractile properties. The nuclei of the endothelial and the connective tissue cells are irregularly spaced and frequently clustered near the free edge of the valve.These ultrastructural features suggest that the function of the lymphatic valves is mainly passive. They are firmly inserted in the lymphatic vessel wall by collagen fibers and their moving parts are slender and elastic. Their endothelium appears relatively impermeable and is firmly attached to the subjacent connective tissue.This study has been supported by a grant from The Council for Tobacco Research—U.S.A.. We thank Professor Robert C. Rosan (Saint Louis University—U.S.A.) for expert advice, R. Janssens for technical, G. Pison and St. Ons for photographic and N. Tyberghien for secretarial assistance.     

Clinical and pathological aspects of filarial lymphedema and its management

Lymphatic filariasis, transmitted by mosquitoes is the commonest cause of lymphedema in endemic countries. Among 120 million infected people in 83 countries, up to 16 million have lymphedema. Microfilariae ingested by mosquitoes grow into infective larvae. These larvae entering humans after infected mosquito bites grow in the lymphatics to adult worms that cause damage to lymphatics resulting in dilatation of lymph vessels. This earliest pathology is demonstrated in adults as well as in children, by ultrasonography, lymphoscintigraphy and histopathology studies. Once established, this damage was thought to be irreversible. This lymphatic damage predisposes to bacterial infection that causes recurrent acute attacks of dermato-lymphangio-adenitis in the affected limbs. Bacteria, mainly streptococci gain entry into the lymphatics through 'entry lesions' in skin, like interdigital fungal infections, injuries, eczema or similar causes that disrupt integrity of skin. Attacks of dermato-lymphangio-adenitis aggravates lymphatic damage causing lymphedema, which gets worse with repeated acute attacks. Elephantiasis is a late manifestation of lymphatic filariasis, which apart from limbs may involve genitalia or breasts. Lymphedema management includes use of antifilarial drugs in early stages, treatment and prevention of acute attacks through 'limb-hygiene', antibiotics and antifungals where indicated, and physical measures to reduce the swelling. In selected cases surgery is helpful.     

Prox1, master regulator of the lymphatic vasculature phenotype

In contrast to the extensive molecular and functional characterization of blood vascular endothelium, little is known about the mechanisms that control the formation and lineage-specific differentiation and function of lymphatic vessels. The homeobox gene Prox1, the vertebrate homologue of the Drosophila prospero gene, has been recently identified to be required for the induction of lymphatic vascular development from preexisting embryonic veins, and studies in Prox1-deficient mice have confirmed Florence Sabin's original hypothesis about the origin of the lymphatic vascular system from embryonic veins. The recent establishment of cell culture models for the selective propagation of blood vascular and lymphatic endothelial cells, together with the findings that these cells maintain their lineage-specific differentiation in vitro, has led to the discovery that Prox1 expression is sufficient to induce a lymphatic phenotype in blood vascular endothelium. Ectopic expression of Prox1 downregulated blood vascular-associated genes and also upregulated some of the known lymphatic endothelial cell markers. Together, these studies suggest that the blood vascular phenotype represents the default endothelial differentiation and they identify an essential role of Prox1 in the program specifying lymphatic endothelial cell fate.     

Lymphatic development

The lymphatic system is essential for fluid homeostasis, immune responses, and fat absorption, and is involved in many pathological processes, including tumor metastasis and lymphedema. Despite its importance, progress in understanding the origins and early development of this system has been hampered by lack of defining molecular markers and difficulties in observing lymphatic cells in vivo and performing genetic and experimental manipulation of the lymphatic system. Recent identification of new molecular markers, new genes with important functional roles in lymphatic development, and new experimental models for studying lymphangiogenesis has begun to yield important insights into the emergence and assembly of this important tissue. This review focuses on the mechanisms regulating development of the lymphatic vasculature during embryogenesis. Birth Defects Research (Part C) 87:222–231, 2009. © 2009 Wiley‐Liss, Inc.     

两种不同淋巴转移能力小鼠肝癌细胞亚系的建立和生物学特性

通过单细胞分离(克隆)培养和局部淋巴结转移灶筛选的方法,建立两种不同淋巴转移能力HepA肝癌细胞亚系,分别命名为HepA-H和HepA-L,并比较其生长、游走和贴壁能力等生物学特性。瘤细胞接种于小鼠足垫后,同侧腘窝淋巴结转移率,HepA-H为83.3%,HepA-L为16.7%,两者具有显著差异(p<0.01)。两种细胞亚系均未见肺、肝、脾、肾等脏器转移灶。体外实验显示,HepA-H的生长、游走和贴壁能力均强于HepA-L。HepA肝癌不同淋巴转移能力亚系的建立,为研究肿瘤经淋巴管转移机理提供了良好的肿瘤淋巴转移动物模型。     

喉淋巴管的几种组织化学显色法比较观察

目的为喉毛细血管和毛细淋巴管的鉴别提供更好的技术方法。方法采用间接注射法,5′-核苷酸酶-碱性磷酸酶双重显色法（5′-Nase-ALPase）,针对基底膜的免疫组化显色法。结果酶组化染色结果显示毛细血管和血管显示有清楚的蓝色轮廓,而附近的毛细淋巴管和淋巴管则呈棕色轮廓,两者易于鉴别。免疫组化染色结果显示毛细血管和血管显示清楚的棕色轮廓,而附近的毛细淋巴管和淋巴管轮廓极其浅淡,两者亦可鉴别。结论酶组化和免疫组化显色法是喉内区别毛细淋巴管和毛细血管的一种可靠且特异的方法。     

建立大鼠持续性肠内营养的输注模型和淋巴液引流方法

目的建立大鼠持续性肠内营养的输注模型和淋巴液引流的方法。方法大鼠行胃造口后游离硅胶管至背部,通过swivel管连接到微量输液泵。5 d持续匀速的输入肠内营养后开腹并游离肠淋巴干,大鼠在肠道缺血60 min再灌注120 min同时潜行插入硅胶管收集淋巴液。结果第一次胃造口术后大鼠持续肠内营养输注通畅,未出现死亡或并发症状。第二次手术时通过较简单的方法可以获取淋巴液,成功率较高,可达90%以上。结论大鼠持续肠内营养输注模型的建立,为研究营养物质的作用搭建了一个平台;通过较简单的方法获取淋巴液可以为研究淋巴液的成分提供便利。     

比较几种组织化学显色法在肺内毛细淋巴管的应用

目的 为肺内毛细血管和毛细淋巴管的鉴别提供更好的技术方法。方法 采用5’-核苷酸酶一碱性磷酸酶双重显色法（5′-Nase-ALP）、针对基底膜的免疫组化显色法及特殊染色法。结果 5′-Nase-ALP显色法能够较客观、较清楚地区分肺内毛细血管和毛细淋巴管。结论 针对基底膜的免疫组化显色法及特殊染色法不够准确或受器官结构特点限制,但对于部分实质性或空腔性器官及肿瘤淋巴管的研究仍不为是一种客观、可靠、经济、实用的方法,值得进一步尝试推广。     

A hypothesis on the role of primitive macrophages in initial embryonic lymphatic development

Recent evidences have shown that macrophages are tightly related to pathological lymphangiogenesis. However, the effects which primitive macrophages take in embryonic lymphatic development remains unclear. Here, we postulate that the primitive macrophages may play an important role in initial embryonic lymphatic development. The possible mechanism: primitive macrophages induce BECs to transdifferentiate into LECs and initiate the budding, moreover, the lymph sacs are not only formed by LECs but also some scattered cells with macrophage characteristics preferentially located in the loose mesenchyme.     

粘附分子在组织内淋巴管内皮和体外培养淋巴管内皮细胞的表达

目的研究粘附分子PECAM-1、ICAM-3、VCAM-1和CD44在组织淋巴管和培养淋巴管内皮细胞(LECs)的表达.方法取狗的空肠和腹前壁皮肤,作冰冻切片,用免疫组织化学染色法标记粘附分子在组织淋巴管内皮的表达.分离和培养狗胸导管的LECs,用免疫荧光染色法标记粘附分子在培养LECs的表达,在共聚焦激光扫描显微镜下观察.用图像分析系统分析粘附分子表达的光密度值(OD值),并与肿瘤坏死因子α(TNF-α)或脂多糖(LPS)刺激细胞的表达进行比较.结果组织内淋巴管内皮的PECAM-1和ICAM-3呈免疫反应阳性.培养LECs表达PECAM-1、ICAM-3和CD44,而VCAM-1的表达不明显.用TNF-α刺激细胞后,PECAM-1和ICAM-3表达的OD值与正常对照组无明显差别,CD44的OD值低于正常对照组.用LPS刺激细胞后,PECAM-1、ICAM-3和CD44表达的OD值均无显著性变化.用TNF-α或LPS刺激LECs后,VCAM-1呈弱表达.结论粘附分子PECAM-1、ICAM-3、VCAM-1和CD44的表达在培养LECs和组织内淋巴管内皮细胞存在着差异.用TNF-α或LPS刺激后,LECs的VCAM-1表达增强.这可能与淋巴管内皮细胞的功能状态有关.