小鼠肝癌细胞移植瘤内淋巴管的形态与亚细胞结构

目的 观察小鼠肝癌细胞所致小鼠移植瘤内淋巴管形态学改变，探讨移植瘤内是否有淋巴管新生。方法 将小鼠肝癌(H22)腹水型瘤株细胞接种于昆明小鼠腋部皮下，2周时摘取肿瘤，采用HE染色观察癌肿的发展，光镜下用5′-核苷酸酶一碱性磷酸酶双重染色法观察毛细淋巴管；半薄切片筛选毛细淋巴管；应用透射电镜观察毛细淋巴管内皮细胞超微结构。结果 动物模型显示2周时小鼠腋部可见明显肿块。5′-核苷酸酶一碱性磷酸酶双重染色法可见中心区无毛细淋巴管，周边部可见稀疏的染成棕黄色的毛细淋巴管，染成蓝色的血管较为多见。透射电镜下，周边区可见毛细淋巴管，亚细胞结构发生损伤。结论 小鼠肝癌细胞所致小鼠移植瘤内存在新生毛细淋巴管。     

吲哚类化合物MAZ51对小鼠移植瘤生长及淋巴管生成的影响

目的:观察吲哚类化合物MAZ51对小鼠移植瘤生长及淋巴管生成的影响,并探讨其作用机制.方法:皮下注射肉瘤(S180)腹水型瘤株构建小鼠移植瘤模型,成瘤后给予MAZ51,观察其对移植瘤生长的影响.免疫组织化学标记淋巴管,分析移植瘤周边区和正常皮肤的毛细淋巴管密度(LMVD).结果:移植瘤周边区的LMVD高于正常皮肤组织.给予MAZ51后,明显减慢移植瘤的生长,但瘤周边区的LMVD并没有变化.结论:实验结果显示肿瘤周边区有淋巴管新生.MAZ51影响移植瘤的生长,但不影响淋巴管的生成.     

小鼠肝癌细胞H22在肿瘤淋巴道转移研究中的应用

目的：探讨小鼠肝癌细胞H22在肿瘤淋巴道转移研究中的应用。方法：将小鼠肝癌细胞株H22分别接种于Km小鼠左腹股沟部（A组）和左后肢爪垫皮下（B组）,于接种后分批处死小鼠。H-E染色观察A组移植瘤及B组胴淋巴结、腹股沟淋巴结;透射电镜下观察A组移植瘤内毛细淋巴管超微结构。结果：A组小鼠移植瘤周边区可见毛细淋巴管,其内皮细胞亚细胞结构发生改变。B组淋巴结发生癌转移的概率和程度与接种细胞数相关。结论：小鼠肝癌（H22）模型可作为一种重要的肿瘤移植性模型而在肿瘤的淋巴道转移研究中发挥作用。     

大鼠原发性结肠癌淋巴管的形态结构

目的　研究大鼠原发性结肠癌毛细淋巴管的微细分布、超微结构特征 ,观察毛细淋巴管的形态结构变化 ,为进一步探讨癌组织的淋巴道转移机制提供形态学依据。方法　取 4周龄大鼠 5 0只 ,用MNNG(甲基硝基亚硝其胍 )灌肠 ,建立大鼠结肠癌模型 ,分期取材 ,应用半薄切片光镜观察、超薄切片电镜观察的方法进行观察。结果　半薄切片观察可见癌组织中心区无毛细淋巴管和淋巴管 ;癌组织周边区毛细淋巴管数量增多 ,管腔扩张 ;电镜观察可见癌组织周边区毛细淋巴管内皮细胞发生溶解破坏 ,内皮细胞间开放连接增多 ,内皮细胞细胞器的形态发生明显改变。结论　癌细胞淋巴道转移可能是由于毛细淋巴管壁的破坏和内皮细胞的开放连接 ,内皮细胞器的改变可能是管壁破坏的基础     

喉癌淋巴管的微细分布和超微结构的观察

目的观察喉癌淋巴管的微细分布和超微结构特征,为探讨喉癌淋巴转移机制提供形态学依据。方法采用5′-核苷酸酶-碱性磷酸酶双重染色法(5′-Nase-ALP)、半薄切片光镜观察、超薄切片电镜观察。结果喉癌组织中心区未见淋巴管,癌周边区和正常区组织内存在毛细淋巴管,与正常区比较,周边区组织内淋巴管数量增多,管腔扩大,形态不规则。淋巴管内皮细胞连接开放增多,并可见部分内皮细胞破裂溶解,管壁不完整,毛细淋巴管内皮细胞变性,细胞器发生明显改变。正常区毛细淋巴管的形态和超微结构未见明显改变。结论喉癌淋巴管的分布及超微结构的改变与喉肿瘤细胞经淋巴道转移密切相关,因而为进一步探讨喉癌颈淋巴结转移的机制以及防治提供了理论依据。     

S180移植瘤瘤周淋巴管和血管光镜观察

目的观察小鼠S180移植瘤及瘤周组织淋巴管和血管的形态学改变.方法将S180小鼠腹水瘤接种于昆明小鼠左腋部皮下,20天切取肿瘤组织及瘤周组织和对侧同一部位正常皮下组织作对照,HE染色观察确认肿瘤.光镜下用Schiff染色观察淋巴管和血管.结果动物模型显示两周以上小鼠腋部可见明显肿块.Schiff染色可见瘤中心区液化坏死,无毛细淋巴管;瘤周边区淋巴管及血管密度和淋巴管及血管检出率明显高于正常侧(P＜0.001～P＜0.005).瘤周边区淋巴管分布不均匀,管腔大,形态不规则,管壁薄.结论S180小鼠移植瘤中心区无淋巴管,可见血管;瘤周边区淋巴管及血管密度高,管腔扩张,形态不规则.     

D-Limonene对小鼠移植瘤生长及淋巴管生成的影响

目的:观察右旋柠烯对小鼠移植瘤生长及淋巴管生成的影响,并探讨其作用机制。方法:皮下注射肉瘤(S180)腹水型瘤株构建小鼠移植瘤模型,给予D-Lim onene干预,免疫组化染色观察瘤细胞V EG F-C表达,LY V E-1标记淋巴管,观察其分布。结果:对照组瘤细胞V EG F-C表达较强,瘤周边部淋巴管较多,有淋巴结、肺转移。用药组瘤细胞V EG F-C表达较弱;瘤周边部淋巴管较少,未见淋巴结、肺转移。结论:D-Lim onene有抑制移植瘤内瘤细胞V EG F-C表达和淋巴管生成的作用,有可能降低肿瘤的淋巴道转移。     

电穿孔法转染GFP标记H22和S180的实验研究

采用电转染法将绿色荧光蛋白(GFP)基因导入小鼠肝癌H22和肉瘤S180细胞,通过G418筛选,建立了稳定表达该蛋白的小鼠肝癌H22细胞株和肉瘤S180细胞株;光镜和电镜观察其细胞形态、超微结构及生长状况没有发生显著改变;建立相应的皮下和腹腔荷瘤动物模型,观察到其皮下和腹腔成瘤时间与腹腔荷瘤生存时间无显著改变(P＞0.05),在活体荧光成像系统能观察到其荷瘤部位荧光.期望利用gfpH22和gfpS180阳性细胞株,运用免疫荧光技术、活体荧光成像系统和激光共聚焦系统对肿瘤进行体内外直观、可视和半定量的深入研究.     

氯喹对S_(180)荷瘤小鼠的抑瘤作用及其机制

目的探讨氯喹(CQ)对小鼠移植性肉瘤S_(180)的抑瘤作用及其可能机制。方法采用40只S_(180)荷瘤小鼠分为模型组,氯喹低剂量组,氯喹中剂量组,氯喹高剂量组进行体内抑瘤实验,观察不同浓度氯喹对小鼠肉瘤S_(180)的抑制作用;透射电子显微镜观察氯喹作用下荷瘤鼠肿瘤细胞超微结构的变化;流式细胞术检测氯喹诱导荷瘤鼠S_(180)细胞凋亡的情况;免疫组织化学方法检测相关凋亡因子Bcl-2、细胞色素C和Cle-Caspase-3的表达情况;Western blotting检测荷瘤组织Bcl-2和Cle-Caspase-3蛋白表达水平的变化以及线粒体和细胞质中细胞色素C蛋白含量的变化。结果与模型组相比较,氯喹处理组小鼠移植肉瘤S_(180)生长速度显著减慢,肿瘤体积和瘤质量明显减小(P0.05);透射电子显微镜观察发现,与模型组比较氯喹处理组肿瘤细胞形态出现明显凋亡损伤改变,凋亡小体形成;各剂量组氯喹均可诱导荷瘤鼠S_(180)细胞凋亡,使抗凋亡因子Bcl-2表达下调,凋亡因子Cle-Caspase-3表达上调(P0.05);并且氯喹能够降低线粒体内细胞色素C蛋白的表达,提高细胞质细胞色素C蛋白的表达(P0.05),促使线粒体内细胞色素C向细胞质内释放。结论氯喹能够抑制小鼠S_(180)肉瘤的生长,可能是通过线粒体凋亡途径诱导细胞凋亡,从而发挥抑瘤的作用。     

小鼠S180移植瘤血管生成作用及其调节机制的研究

目的：观察肉瘤180(S180)移植瘤发展过程中血管生成及血管生成调节因子的变化,并对其调节机制进行探讨。 方法： 利用Km小鼠的S180移植瘤模型，采用FⅧ因子免疫组化染色检测肿瘤血管生成，ELISA和EIA法检测荷瘤鼠肿瘤组织和血浆中血管内皮生长因子（VEGF）和内皮抑制素（endostatin）水平，采用多元回归分析肿瘤组织微血管计数、血管形态与瘤重变化的关系。 结果： 随着荷瘤时间延长，肿瘤组织内微血管计数，瘤内血管相对总量增加，血管的相对面积增大（P<0.05）；肿瘤组织匀浆中VEGF水平在荷瘤10 d、15 d均显著高于5 d组（P<0.05）；endostatin在肿瘤匀浆和血浆中均在荷瘤15 d达到最高（P<0.05）；V/E比值无显著变化；微血管计数、血管相对总面积与瘤重变化有相关性（P<0.01）。 结论： S180移植瘤病期发展中微血管数目增加，血管口径增大，且与瘤重变化呈正相关；肿瘤发展过程中肿瘤局部血管生成正调节因子逐渐增加，促进血管生成；肿瘤局部血管生成调节因子处于相对的平衡。     

香菇多糖对荷瘤小鼠胸腺、脾和肿瘤的影响

目的:探讨香菇多糖不同给药次数对小鼠S180肉瘤的作用及其机制.方法:制备S180荷瘤小鼠动物模型,于接种肿瘤后第2日始分别隔日腹腔注射生理盐水或香菇多糖,按给药3次和7次后分刖处死各组小鼠,观察小鼠胸腺指数、脾指数、肿瘤指数等各项指标以及肿瘤组织的病理改变.结果:给药7次组与同时期荷瘤生理盐水组相比,胸腺指数、脾指数明显增加,且肿瘤生长缓慢,肿瘤重量明显降低,抑瘤率达到47.85%;而给药3次组各项指标的差别与同时期小鼠相比无统计学意义;病理学观察显示香菇多糖给药7次组肿瘤组织中的细胞间质显著增多,瘤细胞周围有较多的淋巴细胞浸润.结论:腹腔注射7次合适剂量的香菇多糖能明显抑制S180荷瘤小鼠肿瘤的生长,其作用机制可能是通过增强荷瘤小鼠自身免疫功能来实现.     

人参皂甙Rh-2对细胞间连接黏附分子在小鼠移植瘤淋巴管表达的影响

目的观察细胞间黏附分子(JAM)在小鼠移植瘤淋巴管内皮细胞的表达及人参皂甙Rh-2的影响,探讨JAM在癌淋巴管转移中的意义。方法于小鼠前肢皮下接种S180小鼠癌性腹水,成瘤后给予人参皂甙Rh-2灌服,6周后取材。用免疫组化法观察LYVE-1、JAM-1、JAM-2在淋巴管内皮细胞的表达。结果在对照组肿块的周边部可见LYVE-1阳性表达的淋巴管,JAM-1、JAM-2在淋巴管有阳性表达,用药组表达减少。结论JAM-1、JAM-2在淋巴管内皮细胞的表达可能与癌淋巴管转移有关。     

Developmental morphology of vascular and lymphatic capillaries in the working myocardium and Purkinje bundle of the sheep septomarginal band

The normal development of vascular and lymphatic capillaries in the right ventricular septomarginal band of the sheep heart was studied in 9 fetuses aged 60-143 days (term = 147 days), 14 lambs aged 1 day to 16 weeks, and 3 adults. Tissue was fixed by perfusion and examined with light and transmission electron microscopy. The septomarginal band is composed of working myocardium and a well-defined peripheral bundle of Purkinje cells. Vascular capillaries of the working myocardium were closely apposed to myocardial cells. By contrast, vascular capillaries of the Purkinje bundle were situated within the connective tissue sheath and septa, at variable distances from the Purkinje cells. After birth, the capillaries of the Purkinje bundle were also found in grooves and tunnels within the Purkinje strands. The ultrastructure of fetal vascular capillaries associated with myocardial and Purkinje cells was initially similar, and characterized by an abundance of synthetic organelles in endothelial cells and pericytes. However, after 115 days in utero, capillary endothelium with diaphragmed fenestrae, 40-60 nm in width, were observed within the Purkinje bundle. The fenestrae attained an average frequency of 1 per 11 capillary cross sections just before term, and this was maintained in lambs and adults. The ultrastructure of lymphatic capillaries, which were not observed in the septomarginal band until just before term, changed little during development.     

甲壳胺抗肿瘤的实验研究

目的研究甲壳胺对S180荷瘤小鼠的抗瘤作用。方法取S180荷瘤小鼠40只随机分为实验组和对照组,实验组进行不同剂量(50、100、200mg/(kg·d))甲壳胺灌胃处理,对照组仅灌胃生理盐水,连续10d。灌胃结束时检测两组小白鼠的体重变化,观察甲壳胺对腹水瘤小鼠腹水生成的影响,解剖腹腔观察腹水瘤生长情况,HE染色观察瘤细胞。结果实验组小白鼠体重大于对照组小鼠体重,腹水量减少,解剖结果显示,实验组腹水瘤块小于对照组,三种剂量的抑瘤率分别是17.4%、38.8%和41.6%。通过对细胞状态的比较,实验组的瘤细胞数目减少,瘤细胞的体积小,变形、死亡细胞增多,核分裂较少见。结论甲壳胺在体内具有抗S180腹水瘤形成的作用。     

Morphological and Cytogenetic Studies of a Human Synovial Sarcoma Xenotransplanted into Nude Mice

Specimens of a synovial sarcoma from the left thigh of a 60 year old woman and cells from the tumor transplanted into nude mice were examined morphologically and immunohistochemically, and the karyotype was analyzed. lmmunohistochemically, cells of both the parent tumor and the transplanted tumor were positive for vimentin and cytokeratin. The multicystic features of the parent tumor were not reproduced in the mice. On ultrastructural examination, the parent tumor cells were found to have dilated channels of endoplasmic reticulum as well as a junctional apparatus, and they formed extracellular spaces. The transplanted tumor cells, in contrast, had a poorly developed endoplasmic reticulum and were devoid of extracellular spaces. Chromosome analysis of the tumor cells revealed a translocation, t (X; 18) (p 11; q11), as reported previously for synovial sarcoma, thus suggesting the diagnostic utility of the chromosome pattern for identification of synovial sarcoma.     

Morphological and cytogenetic studies of a human synovial sarcoma xenotransplanted into nude mice

Specimens of a synovial sarcoma from the left thigh of a 60-year-old woman and cells from the tumor transplanted into nude mice were examined morphologically and immunohistochemically, and the karyotype was analyzed. Immunohistochemically, cells of both the parent tumor and the transplanted tumor were positive for vimentin and cytokeratin. The multicystic features of the parent tumor were not reproduced in the mice. On ultrastructural examination, the parent tumor cells were found to have dilated channels of endoplasmic reticulum as well as a junctional apparatus, and they formed extracellular spaces. The transplanted tumor cells, in contrast, had a poorly developed endoplasmic reticulum and were devoid of extracellular spaces. Chromosome analysis of the tumor cells revealed a translocation, t (X; 18) (p11; q11), as reported previously for synovial sarcoma, thus suggesting the diagnostic utility of the chromosome pattern for identification of synovial sarcoma.     

A fibrillar elastic apparatus around human lymph capillaries

Summary A fibrillar elastic apparatus around the wall of human lymph capillaries is demonstrated by means of histochemical and ultrastructural techniques. This apparatus consists of three interlinked components listed here in order of increasing distance from the capillary wall: 1) oxytalan fibres connected to the abluminal surface of the endothelial cells, known also as anchoring filaments and consisting of bundles of microfibrils; 2) elaunin fibres consisting of microfibrils and a small amount of elastin; and 3) typical elastic fibres consisting of microfibrils and abundant elastin.The microfibrillar constituent has similar ultrastructural features in the three components of the elastic apparatus. Microfibrils have a diameter of 12–14 nm, an electrontransparent core and a wall with 3–5 electron-dense subunits and oblique cross striations with a period of 15–17 nm. Microfibrils are the common element of the three components of the elastic apparatus and they link them to one another and to the elastic network of the perivascular connective tissue. An elastic apparatus was not found around blood capillaries and it can thus provide a histological marker to identify lymph capillaries. The possible role of the lymphatic elastic apparatus in the physiological activity of the lymphatic absorbing network is discussed and it is proposed that its disconnection from the elastic network of the tissue may promote pathological conditions such as lymphoedema or diseases related to impaired immune responses.     

Pathological Analysis of Cell Differentiation in Cholesterol Granulomas Experimentally Induced in Mice

In this study, cholesterin was implanted in the subcutaneous tissue in mice to induce the formation of cholesterol granuloma. Histological examination was carried out to determine the type and source of cells. The tissue surrounding the embedded cholesterin was examined histologically within the period of 6 months. Cell differentiation in cholesterol granulomas was investigated using ddY mice and GFP bone marrow transplanted mice. Cholesterin was embedded in mice subcutaneously and histopathological examination was carried out in a period of 6 months. Results showed that at 2 weeks, cholesterin was replaced partly by granulation tissues. The majority of cells in the granulation tissues were macrophages and foreign body giant cells and the center consists of small amount of fibroblasts, collagen fibers and capillaries. At 3 months, more granulation tissue was observed compared to 2 weeks. Similar cells were observed, however, there were more fibroblasts, collagen bundles and capillaries present compared to 2 weeks. At 6 months, the cholesterin was mostly substituted by fibrous tissues consisting mainly of fibroblasts and collagen fibers with some macrophages and foreign body giant cells. Specifically, the outer part of the tissue consists of fibroblasts, collagen bundles and capillaries and the inner portion is filled with collagen bundles. Immunohistochemistry revealed that macrophages and foreign body giant cells were positive to GFP and CD68 although the fibroblasts and capillaries in the outer portion of cholesterol granulomas were GFP negative. Some spindle shape fibroblasts were also GFP positive. Immunofluorescent double staining revealed that cells lining the blood vessels were both positive to GFP and CD31 indicating that those were endothelial cells and were actually derived from the transplanted bone marrow cells. The results suggest that macrophages, foreign body giant cells as well as fibroblasts and capillary endothelial cells are bone marrow derived mesenchymal cells.     

大鼠胃壁淋巴管形态分布的实验研究

本文用光镜和透射电镜观察了饱食与饥饿状态下大鼠胃壁淋巴管的微细分布、出现率和形态结构的特点，结果表明：两组大鼠胃壁内淋巴管的出现率及在各层内的分布均无显著差异。粘膜层仅见毛细淋巴管，其它各层可见毛细淋巴管和淋巴管。光镜下饱食鼠胃壁内毛细淋巴管和淋巴管的管腔较大且饱满，而饥饿鼠相同区域的毛细淋巴管和淋巴管的管腔较小，形态更加不规则。电镜下也可见到饱食鼠毛细淋巴管的管壁较平整，皱褶与质膜突起较少，而饥饿鼠毛细淋巴管壁较厚，皱褶和质膜突起较多。     

Ultrastructure of hepatic hemangiopericytoma in the medaka (Oryzias latipes)

The histologic and ultrastructural features of hepatic hemangiopericytoma from a medaka (Oryzias latipes) exposed for 48 hr to 400 mg/liter of diethylnitrosamine at 14 days of age are described. The predominant histologic pattern was of spindle-shaped cells forming numerous whorls around central capillaries, vacuolated areas, or necrotic debris. The predominant cell type was a spindle-shaped cell with oval nuclei, elongated cell processes, and abundant organelles converging upon normal appearing capillaries. Occasionally, however, they converged upon cells swollen with cytoplasmic filaments and/or containing large fenestrated or debris-filled cytoplasmic vacuoles. These features were reminiscent of endothelial cells undergoing intracellular canalization seen in angiogenesis or neovascularization. Individual capillaries were also seen in the mass independent of whorls. It was not clear, as is the case in man, if capillary formation was an integral part of the neoplastic process or a reactive response. Although the liver is an unusual location for hemangiopericytoma in man, many of the cellular features in the fish tumor were similar to the human tumor. The ultrastructural characterization of tumor cells in fish carcinogenesis correlated with histologic patterns of growth will expand our understanding of how fish cells respond when transformed, and augment the development and use of aquatic bioassays for carcinogenesis research.