iNOS抑制剂SMT对内毒素血症大鼠淋巴管动力学的影响

目的 :研究一氧化氮 (NO)在急性内毒素血症中对淋巴管运动的调节作用及诱生型一氧化氮合酶 (iNOS)抑制剂硫酸甲基异硫脲(SMT)对内毒素血症中淋巴微循环的作用。方法 :用倒置显微镜和图像处理系统对肠系膜淋巴管进行动态观测 ,并用硝酸还原酶法测定血清中NO的含量。结果 :LPS组肠系膜淋巴管管径比正常扩大 ,运动频率降低 ,淋巴管运动指数下降。LPS +SMT组则有显著改善。随时间延长LPS组血清NO含量显著升高 ,LPS +SMT组则有明显下降。结论 :内毒素血症中NO过量产生 ,使肠系膜淋巴管扩张 ,收缩减弱。内毒素血症早期应用iNOS抑制剂SMT可维持淋巴微循环于正常水平。     

山莨菪碱对内毒素血症大鼠肠系膜血管内皮细胞与白细胞粘附力的影响

本文采用超高速摄录相和荧光显微系统装置，检测了活体大鼠肠系膜微循环中同一微血管内的白细胞流态和血流速度的变化，观察了山莨菪碱对内毒素作用下的微静脉内皮细胞与细胞粘附反应增强的影响。     

糖皮质激素受体阻断对大鼠内毒素血症时肠系膜微循环变化的影响

以Ru_(486)阻断糖皮质激素受体(GR),观察了静脉注射内毒素引起的肠系膜微循环变化以及地塞米松预处理的保护作用。实验结果提示:1.内毒素血症时内源性糖皮质激素就有通过其受体介导的微循环保护作用;2.糖皮质激素不仅在应激情况下,即使在正常情况下可能就有抑制白细胞活化作用;3.大剂量糖皮质激素对内毒素休克有一定的防治作用,这一作用至少部分地是由GR所介导的。     

急性内毒素血症早期大鼠淋巴管运动变化及其一氧化氮合酶组织学研究

目的:研究急性内毒素血症早期大鼠淋巴管运动变化和一氧化氮合酶(NOS)的相关组织染色,探讨二者的关系。方法:股静脉注脂多糖(LPS),观察大鼠肠系膜淋巴管2h内的运动变化及相关组织的酶组化、HE染色。结果:LPS组淋巴管管径变大,运动频率减少,镜下见小肠绒毛中央乳糜管及粘膜下淋巴管丛极度扩张;对照及LPS组淋巴管内皮细胞及管壁上神经纤维都含有NOS阳性反应产物。结论:内毒素血症早期淋巴管扩张,运动减少,NOS可能在其中起重要作用。     

肠系膜淋巴管结扎对大鼠急性肺损伤的影响

目的：探讨结扎肠系膜淋巴管对失血-脂多糖（LPS）致大鼠急性肺损伤（ALI）的拮抗作用。方法：雄性Wistar大鼠45只，均分为结扎组、未结扎组、假手术组，以失血、LPS复制二次打击模型，结扎组行肠系膜淋巴管结扎术致肠淋巴液断流。在手术创伤后24 h，所有大鼠颈总动脉放血，进行血气分析；从左肺收集支气管肺泡灌洗液（BALF），观察WBC、NO及其合酶、SOD、MDA以及肺泡通透性指数等指标的水平；右肺制备10%组织匀浆，检测MPO、ATPase活性等指标；观察右肺后叶超微结构。结果：二次打击后，未结扎组动脉血PaCO2、BALF中细胞总数及PMN、NO2-/NO3-、NOS、MDA含量以及肺匀浆MPO活性、肺通透性指数均显著高于假手术组，动脉血pH、PaO2、BALF中SOD、肺匀浆ATPase活性显著低于假手术组（P<0.01，P<0.05）；结扎组大鼠BALF中细胞总数及PMN、MDA、NO2-/NO3-含量、肺通透性指数均显著高于假手术组，BALF中SOD活性显著低于假手术组（P<0.01，P<0.05）。但结扎组大鼠动脉血pH、PaO2、肺匀浆ATPase活性显著高于未结扎组，动脉血PaCO2、BALF中细胞总数及PMN、NO2-/NO3-、NOS、MDA含量、肺通透性指数及肺匀浆MPO显著低于未结扎组（P<0.01，P<0.05）；且肺血管内皮细胞损伤程度较未结扎组轻微。结论：肠系膜淋巴管结扎可减轻失血-LPS致大鼠的急性肺损伤。提示二次打击的肠系膜淋巴液在大鼠急性肺损伤的发病过程中发挥着重要作用。     

牛珀至宝微丸对内毒素休克肾组织诱生型一氧化氮合酶表达的影响

目的　探讨牛珀至宝微丸对内毒素休克肾组织iNOS表达的作用。方法　静脉注射内毒素 (LPS) 1 5mg/kg、腹腔注射D 氨基半乳糖糖 (D Ga1N) 10 0mg/kg造成内毒素休克模型 ,用牛珀至宝微丸干预处理 ,免疫组化方法检测诱生型一氧化氮合酶 (iNOS)在肾组织内的表达。结果　牛珀至宝微丸可以使肾内iNOS表达减弱 ,肾损伤减轻。结论　牛珀至宝微丸可以减轻内毒素休克造成的肾损伤 ,这种保护作用可能是通过调节肾组织NOS表达而产生的     

一氧化氮合酶抑制剂N-硝基-L-精氨酸甲基酯对大鼠淋巴管运动的影响

目的研究一氧化氮合酶抑制剂N-硝基-L-精氨酸甲基酯对大鼠肠系膜淋巴管作用。方法股静脉注入N-硝基-L-精氨酸甲基酯,活体动态观察其2小时内淋巴管运动的变化;用HE染色、酶组化染色研究其组织学改变。结果发现静注N-硝基-L-精氨酸甲基酯可使淋巴管管径变细,运动频率增加,L-精氨酸可暂时逆转该变化;镜下发现淋巴管内皮细胞及管壁上神经纤维都含有一氧化氮合酶阳性反应产物,注N-硝基-L-精氨酸甲基酯组小肠组织HE切片出现固有层中央乳糜管扩张。说明一氧化氮合酶在淋巴循环中可能具有重要调节作用。     

急性内毒素血症早期大鼠肠系膜淋巴管的动态观察

目的利用活体微循环显微闭路电视系统,研究内毒素作用下肠系膜淋巴管的运动变化,探讨内源性NO对淋巴管的作用.方法Wistar大鼠分三组,于股静脉一次性推注:(1)内毒素(脂多糖,1ipopolysaccharide,LPS);(2)一氧化氮合酶(NOS)抑制剂N-硝基-L-精氨酸甲基酯(L-NAME);(3)LPS及L-NAME.观察注药前和注药后2 h内的肠系膜淋巴管管径、收缩频率等指标.结果(1)LPS组:肠系膜淋巴管管径较注药前扩大,运动频率减少,总收缩活性指数变小;(2)L-NAME组:淋巴管管径变细、运动频率增加、收缩分数变大;(3)LPS L-NAME组:淋巴管管径、运动频率、运动指数2 h内变化不显著.结论内毒素血症早期肠系膜淋巴管扩张、收缩减弱,同时应用L-NAME可抑制该变化,单独应用L-NAME则使淋巴管缩窄、收缩加强,提示NO可能在维持正常淋巴管运动及内毒素血症早期淋巴循环变化过程中起重要作用.     

内毒素血症大鼠肠系膜淋巴循环动力学及iNOS抑制剂SMT作用的实验研究

本实验主要研究内毒素、iNOS抑制剂硫酸甲基异硫脲(SMT)对活体淋巴管运动及iNOS的表达的影响,探讨iNOS、iNOS抑制剂SMT在急性内毒素血症中对淋巴微循环的作用。 动物选用雄性Wistar大鼠54只,体重200-300g。将动物分为3组control组(n＝18)股静脉一次推注生理盐水(5mL／kg体重)作对照。分别在注药后l、3、6h各处死6只;LPS组(n＝18)股静脉一次推注LPS(5mg／kg体重),其它处理同control组;LPS+SMT组(n＝18)股静脉一次推注LPS(5　mg／kg体重)和SMT(10　mg／kg体重);其它处理同control组。 动物用20％乌拉坦(7ml／kg体重)肌肉注射麻醉。腹正中部开腹,取出肠系膜,找出一条理想的游离淋巴管,置于观察盒中,保温(32℃左右),并定时于肠管上滴加37℃左右的台氏液以维持一定湿度,在倒置显微镜下观察并录像。将每只需观察6h的动物镜下先稳定5-10min,作为正常录像,再按分组注药,观察录像6h后,处死。 根据录像资料,利用微机图像处理系统,定淋巴管、定部位测量实验用药前及用药后l、2、3、4、5、6h各时段各动物淋巴管管径的大小,求得平均口径(d)、最大舒张口径(b)、最小收缩管径(c)、淋巴管收缩频率(a),计算淋巴管运动指数［收缩指数IndexI＝(b2-c2)／b2,总收缩活性指数IndexII＝(b2-c2)a／b2,淋巴管动力学指数LD-Index＝(b-C)100a／d2］。 所有动物在处死后,取血静置,取血清,用一氧化氮试剂盒测定血清中NO合量。并取肠系膜进行免疫组化染色。 结果(1)LPS组的收缩频率、运动指数均比注药前显著降低(P＜0.05),免疫组化染色阳性细胞数明显增多(P＜0.05)。阳性细胞为巨噬细胞和内皮细胞。血浆NO合量显著升高(P＜0.01)。(2)SMT+LPS组的收缩频率、运动指数均与注药前相比,无显著差异(P＞0.05)。免疫组化染色阳性细胞数及血浆NO含量与LPS组相比,均显著降低(P＜0.01)。 结果说明正常条件下,肠系膜淋巴管运动不但受自身节律的调节,还受NO的调节。LPS作用下,肠系膜淋巴管随时间延长明显扩张,运动减弱,渗出增多,与NO的生成量呈正相关。SMT能抑制iNOS的活性,内毒素血症早期应用选择性iNOS抑制剂,可维持淋巴循环于正常水平。内毒素血症中,由于iNOS过度表达,导致过量的NO产生,使淋巴管的运动异常。SMT由于能抑制iNOS,从而抑制了NO的过量产生,使淋巴管运动恢复。这可能为临床上淋巴管循环障碍及内毒素血症的治疗提供理论基础和实验依据。     

诱生型一氧化氮合酶的变化在高胆固醇血症大鼠肾损害中的作用

目的:探讨高胆固醇血症大鼠肾组织诱生型一氧化氮合酶(iNOS)的变化与脂质肾损害的关系。方法:采用生化法检测血脂、尿蛋白、尿及肾皮质中一氧化氮含量,免疫组化法及逆转录—多聚酶链法检测肾组织中iNOS的表达强度及水平。TUNEL法检测肾组织中凋亡细胞。结果:8周时高脂组大鼠血清总胆固醇、低密度脂蛋白胆固醇、24h尿蛋白定量及尿一氧化氮量、肾皮质一氧化氮含量明显高于对照(P＜0.05)。肾皮质iNOSmRNA表达上调(P＜0.01)。肾皮质一氧化氮量与24h尿蛋白定量呈正相关(P＜0.05)。肾皮质iNOS积分光度与肾组织细胞凋亡指数呈正相关(P＜0.05)。肾组织细胞凋亡指数与24h尿蛋白含量呈正相关(P＜0.01)。结论:iNOS表达增强与脂质肾损害有关,可能通过引起细胞凋亡导致脂质肾损害。     

Higher blood vessel density in comparison to the lymphatic vessels in oral squamous cell carcinoma

Introduction: Oral squamous cell carcinoma (OSCC) is characterized by local invasion and the development of cervical metastasis. In the tongue, an association between the invasion of the lymphatic vessels and the development of metastasis in the regional lymph nodes has been demonstrated. Moreover, invasion of the blood vessels is associated with greater recurrence and poorer prognoses. Therefore, the presence and density of lymphatic and blood vessels in intra- and peritumoral tissues should play an important role in the progression, dissemination and metastasis of carcinomas. However, the evidence regarding OSCC is inconclusive. The aim of this study was to determine the comparison and association between the lymphatic (D2-40) and blood vessel (CD34) densities in intratumoral OSCC tissue. Materials and Methods: Thirty-seven cases diagnosed as OSCC between the years 2000 and 2008 were obtained from the Anatomic Pathology Service of the School of Dentistry, University of Chile. The immunohistochemical markers D2-40 and CD34 were used, and the densities (mm²) of lymphatic vessels (LVD) and blood vessels (BVD) in the intratumoral region were determined. The relationship between LVD and BVD values was evaluated. Results: There were significant association between the CD34 and D2-40 expression (rho=0.4, P<0.05) and between the LVD and the location in the tongue (P=0.019). The BVD was greater (128.0 vessels/mm²) than the LVD (42.9 vessels/mm²), and there was a positive correlation between the LVD and BVD. Conclusions: In OSCC, the BVD is greater than the LVD, and there is a moderate correlation between the two quantities.     

淋巴管阻断对大鼠睾丸影响的自动图像分析

目的 :研究淋巴管阻断后对睾丸的定量组织学影响。方法 :用电灼法将双侧睾丸淋巴管阻断 ,对双侧睾丸淋巴管阻断术后 2天、6天、1 2天、2 4天和 48天的 2 5只大鼠进行研究。另 2 5只大鼠为假手术对照。将睾丸组织切片输入TAS plus型自动图像分析仪进行定量组织学测定。结果 :在睾丸淋巴管阻断术后 6天、1 2天和 2 4天 ,睾丸间质的面积占总面积的比值增加 ,而生精小管的面积占总面积的比值则减少。在睾丸淋巴管阻断术后 2天、6天、1 2天、2 4天和 48天 ,生精上皮的面积占总面积的比值均减少 ,而间质面积与生精上皮面积的比值则均增加 ,生精小管的直径和生精上皮的厚度均变小。结论 :以上结果表明睾丸淋巴管阻断对生精上皮的影响比对间质的影响更为明显。     

The morphology of the human lymphatic vessels in the head and neck

Previously little has been written about the morphology of the human lymphatic vessels since Sappey (Sappey [ 1874 ] Anatomie, Physiologie, Pathologie des Vaisseaux Lymphatiques, Paris: Adrien Delahaye) over 100 years ago. There needs to be an accurate re‐evaluation of scientific observations to aid clinical management. Forty‐nine combinations of tissue from the head and neck of 20 unembalmed human cadavers were studied. Six percent hydrogen peroxide was used to find the vessels. They were injected with radio‐opaque mixture, dissected, photographed, and radiographed. Final results were transferred to the computer for analysis. Different sized lymphatic valves were found in the precollecting and collecting lymph vessels, the lymphatic trunks, and ducts. The intervals between the valves were of various lengths. Diverse lymphatic ampullae and diverticula were seen in precollecting and collecting lymph vessels. Initial lymph vessels arose from the dermis, the galea, and the mucosal membrane. The vasculature of the direct and indirect precollecting and collecting lymph vessels, lymphatic trunks, and ducts was recorded. The morphology of the human lymphatic vessels in the head and neck has been described and recorded using radiographs and photographs. Clin. Anat. 23:654–661, 2010. © 2010 Wiley‐Liss, Inc.     

Blood supply to the lymphatic vessels in the leg: An incidental finding

There are no reports or images of the blood supply to the lymphatic vessels. One lower limb of an unembalmed human cadaver was studied. Hydrogen peroxide (6%) was applied to find the lymphatic vessels by using a surgical microscope. The vessels were injected with a radio‐opaque mixture and dissected. During the dissection, several sites of paralymphatics arteriole nutrient (PAN) vessels were found in close proximity to collecting lymphatic vessels in the medial aspect of the leg. The caliber of the lymphatic vessels was about 1 mm. The caliber of PAN vessels was <0.1 mm. The blood vessels were seen running along the lymphatic vessels. Some of them crossing the lymphatics and supplying the fatty tissue nearby and some running parallel on the lymph vessel walls. Histology sections show different‐sized PAN vessels containing blood cells situated close to the lymphatic wall and within the lymphatic vessel wall. PAN vessels have been found and described. It will upgrade our anatomical knowledge and also be of benefit for medical and/or scientific research. Clin. Anat. 23:451–454, 2010. © 2010 Wiley‐Liss, Inc.     

Development of the peritoneal lymphatic stomata and lymphatic vessels of the diaphragm in mice

The generation and development of the peritoneal lymphatic stomata (PLS) and lymphatic vessels of the diaphragm were studied in mice at gestational ages from the embryonic to the postnatal period with TEM, SEM and enzyme histochemistry and the PLS data were quantitatively analyzed with computer-assisted image processing technology (Elescope image analysis software). The results showed that the diaphragmatic mesothelium was covered only by flattened mesothelial cells (FMC) at the 13th embryonic day (ED 13). At ED 15, some cuboidal mesothelial cells (CMC) and immature lymphatic stomata (NLS) were found scattered on the diaphragmatic mesothelium. The sub-peritoneal lymphatic capillaries did not appear until ED 18. However, no absorptive function was observed in NLS when trypan blue granules were injected into the peritoneal cavity. At postnatal day 1 (PND 1), the endothelial cytoplasm processes of the diaphragm lymphatic capillaries span the connective tissue fibers and the basal membrane of CMC to form the subperitoneal channels. These channels were connected with NLS and serve as the absorptive route between the peritoneal cavity and the sub-peritoneal lymphatic vessels. The trypan blue absorption test demonstrated that postnatal PLS possessed an absorptive function and had transformed to mature lymphatic stomata (MLS) by PND 1. Thus, NLS were renamed of MLS. At PND 5, the cuboidal mesothelial cell ridge (CMCR) appeared with increased CMC areas. At PND 10, CMCR were fused to form the band-like CMC area with much more MLS distributed in the muscular portion of the diaphragm. With distribution area and density of PLS increasing and growth of lymphatic vessels, an increased absorptive function from the peritoneal cavity was observed in the experiment.     

磷脂酰肌醇特异性磷脂酶C对内毒素血症大鼠肝组织CD14表达的抑制作用

目的观察磷脂酰肌醇特异性磷脂酶C(PI—PLC)对大鼠内毒素血症时肝组织中Kuiter细胞(KCs)的活性、CD14 mRNA的表达和血浆细胞因子释放的作用。方法将Wistar大鼠90只,随机分为LPS组(静注LPS 5mg／kg)、PI—PLC组(注LPS前30min注PI-PLC 100U／kg)和生理盐水NS组。分别于注射前及注射后1、3、6和12h取材,每组每时相点6只,测定血浆内毒素(鲎试剂基质偶氮显色法)、LBP及TNF-α(均用ELISA法)和IL-6(放免法)的含量,用RT-PCR检测肝组织中CD14 mRNA的表达,并观察其形态学变化。结果LPS组LBP、TNF-α、IL-6的含量和CD14mRNA的表达明显增加,并伴有KCs激活,数量增多,体积增大,吞噬功能增强,肝细胞出现变性和坏死等;而PI-PLC处理组所致的上述变化明显减轻。结论PI—PLC对内毒素所致肝组织内KCs的激活有一定抑制作用。     

宫颈癌周边区淋巴管形态学观察及其意义

目的:观察人宫颈癌周边区淋巴管的形态、分布特点,探讨癌淋巴道转移机制。方法:将临床手术切除的宫颈癌组织,按癌组织中心区、周边区及正常区取材,采用LYVE-1免疫组化法、半薄切片光镜观察法,对宫颈癌淋巴管的形态分布和癌组织进行了形态学观察。结果:光镜下可见癌细胞已突破基底膜并向间质浸润。宫颈正常区粘膜层缺少淋巴管,癌周组织有较多的淋巴管,并可见淋巴管扩张,管壁模糊破损。统计癌周边区微淋巴管的面密度(0.25±0.06)、数密度(7.97±1.88)的结果表明均高于正常区(0.13±0.04)和(3.50±0.52()P<0.01)。结论:宫颈癌周边区淋巴管数量和形态的改变,为癌细胞进入淋巴管提供了形态学基础。     

Murine Notch1 is required for lymphatic vascular morphogenesis during development

BACKGROUND: The transmembrane receptor Notch1 is a critical regulator of arterial differentiation and blood vessel sprouting. Recent evidence shows that functional blockade of Notch1 and its ligand, Dll4, leads to postnatal lymphatic defects in mice. However, the precise role of the Notch signaling pathway in lymphatic vessel development has yet to be defined. Here we show the developmental role of Notch1 in lymphatic vascular morphogenesis by analyzing lymphatic endothelial cell (LEC)‐specific conditional Notch1 knockout mice crossed with an inducible Prox1CreER^T2 driver. RESULTS: LEC‐specific Notch1 mutant embryos exhibited enlarged lymphatic vessels. The phenotype of lymphatic overgrowth accords with increased LEC sprouting from the lymph sacs and increased filopodia formation. Furthermore, cell death was significantly reduced in Notch1‐mutant LECs, whereas proliferation was increased. RNA‐seq analysis revealed that expression of cytokine/chemokine signaling molecules was upregulated in Notch1‐mutant LECs isolated from E15.5 dorsal skin, whereas VEGFR3, VEGFR2, VEGFC, and Gja4 (Connexin 37) were downregulated. CONCLUSIONS: The lymphatic phenotype of LEC‐specific conditional Notch1 mouse mutants indicates that Notch activity in LECs controls lymphatic sprouting and growth during development. These results provide evidence that similar to postnatal and pathological lymphatic vessel formation, the Notch signaling pathway plays a role in inhibiting developmental lymphangiogenesis. Developmental Dynamics 243:957–964, 2014. © 2014 Wiley Periodicals, Inc.     

腹腔镜胰十二指肠切除术中肠系膜上血管的应用解剖

目的:为腹腔镜胰十二指肠切除术(laparoscopic pancreatoduodenectomy,LPD)中探查、显露、分离肠系膜上血管提供解剖学依据。方法:10例已固定成人尸体腹部标本,2例新鲜成人尸体。全组男7例,女5例。进行解剖观测,并行模拟腹腔镜操作。结果:⑴肠系膜上静脉(superior mesenteric vein,SMV)的十二指肠水平部段长(3.80±0.72)cm、胰头钩突部段长(1.76±0.25)cm、胰颈后段长(3.81±0.64)cm、胰颈上段长(4.73±1.31)cm,其中胰头钩突部段属支最多;⑵胃网膜右静脉汇入SMV有6种类型;⑶肠系膜上动脉(superior mesenteric artery,SMA)距腹腔干下方(1.12±0.15)cm起自腹主动脉前壁,主干长(3.97±0.54)cm,外径(0.69±0.03)cm,胰十二指肠下动脉和第1空肠动脉起源SMA的有5种类型。结论:(1)SMV的胰头钩突部段最短,属支最多,显露分离最难;(2)LPD中以胃网膜右静脉为标志来探查、显露SMV较好;(3)对肠系膜上动、静脉的显露分离应采用不同的主操作孔来进行;(4)充分利用腹腔镜的放大作用和超声刀的精确切割特性是可以探查、显露和分离好肠系膜上血管的。     

人胰腺癌淋巴管的分布及形态观察

目的观察人胰腺癌淋巴管的分布及形态结构,探讨胰腺癌淋巴道转移机制。方法取手术后人胰腺癌标本21例,应用免疫组化染色法LYVE-1标记淋巴管进行淋巴管计数,半薄切片光镜观察和超薄切片透射电镜观察胰腺癌组织淋巴管的形态及分布特点。结果胰腺癌组织中LYVE-1染色阳性的脉管具有淋巴管的形态学特征,可见癌周组织的微淋巴管数量较癌旁"正常区"有所增加(P<0.01);半薄切片光镜下可见癌周边区和"正常区"淋巴管存在,癌中心区未见有淋巴管;电镜下癌周边区淋巴管内皮细胞连接开放,部分内皮细胞破裂溶解,管壁不完整。淋巴管内皮细胞的线粒体、高尔基体等细胞器改变。结论胰腺癌组织淋巴管主要位于癌周围浸润区的纤维结缔组织中,且淋巴管数量较癌旁"正常区"增多,淋巴管内皮超微结构改变。胰腺癌淋巴管转移可能通过增多的淋巴管的内皮连接开放和对内皮细胞的破坏溶解作用进入淋巴管管壁。