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[Study on distribution and drainage of lymphatic vessels of tongue].

OBJECTIVE: To investigate the distribution and drainage of lymphatic vessels of tongue, and to provide anatomical evidence for treatment of tongue cancer. METHODS: Indirect lymphatic injection was employed, combined with clearing method with winter green oil and corrosive cast technique, to study the distribution of lymphatic vessels of tongue. Anatomical methods were used to detect the sentinel lymph nodes in different region of tongue. RESULTS: The lymphatic vessels of dorsal mucosa composed of lymphocapillary vessels and anstomosing side branches were present by superficial and deep capillary networks. The distribution of lymphatic networks extend from tip to base and from one board to another, and was not influenced by the sulcus tenninalis and median lingual sulcus. Lymphatic vessels in the muscular portion communicated with lymphocapillary network of dorsal and ventral mucosa, which made the lymphatic vessels of tongue to be an integrity network structure. These characters of distribution influenced the lymphatic drainage of tongue. The results showed principal sentinel lymph nodes (SLNs) for anterior part of tongue were submental lymph nodes, submandibular lymph nodes and juguloomohyoid lymph nodes, for lateral part and middle part of tongue were submandibular lymph nodes, jugulodigastric lymph nodes and thyroid lymph nodes, and for root part of tongue were jugulodigastric lymph nodes. SLNs for every injection region were all presented at bilatral neck, but the frequency of stained SLNs at homolateral neck was more than that at contralateral neck. CONCLUSION: The lymphatic vessels of tongue arranged like a network, which made the lymphatic drainage at various ways and made the distribution of sentinel lymph nodes to be bilateral and dispersive.

Drainage↗

Proteinase-activated receptor 2 activation modulates guinea-pig mesenteric lymphatic vessel pacemaker potential and contractile activity.

Lymphatic vessels rhythmically constrict to avoid fluid and protein accumulation in the interstitial space. This activity is critical during inflammation to prevent excessive oedema. Lymphatic pumping is intrinsic to the smooth muscle in the vessel wall and is due to the spontaneous occurrence of action potentials, the pacemaker of which is proposed to be spontaneous transient depolarizations (STDs). This function is highly susceptible to the fluid load and modulated by chemical agents, amongst which inflammatory mediators are important players. Activation of proteinase-activated receptors (PARs) has been involved in inflammation and affects vascular smooth muscle tone. The present study aims to investigate the role of PAR2, a member of the PAR family, in lymphatic vessel pumping. RT-PCR experiments revealed that PAR2 message is present in lymphatic vessels of the guinea-pig mesentery. Agonists of PAR2 such as trypsin and the activating peptide, SLIGRL-NH2, caused a decrease in the contractile activity of intraluminally perfused lymphatic vessels. Moreover, intracellular microelectrode recordings from isolated vessels revealed that PAR2 activation hyperpolarized the lymphatic smooth muscle membrane potential and altered STD amplitude and frequency. The decreases in constriction frequency and STD activity as well as the hyperpolarization were dependent on a functional endothelium, not affected by NO synthase or guanylyl-cyclase inhibition, but mimicked by PGE2 and iloprost and blocked by indomethacin (10 microM) and glibenclamide (1 microM). These results show that PAR2 activation alters guinea-pig lymphatic vessel contractile and electrical activity via the production of endothelium-derived cyclo-oxygenase metabolites.

Animals↗

Immunohistochemical differentiation between lymphangiographically verified lymphatic vessels and blood vessels.

In order to investigate the antigen profile in human lymphatic vessels when compared with blood vessels, postmortem retrograde lymphangiography was done via the thoracic duct on six patients. Formalin fixed, paraffin embedded tissue was stained immunohistochemically for Factor VIII-Related antigen (F VIII R:Ag), with Ulex Europaeus 1 lectin (UEA-1) and for laminin. The results show that the endothelium of blood vessels and lymphatics at all levels of the lymphatic system react positively following staining for Factor VIII-R:Ag and with UEA-1 lectin. The staining for F VIII R:Ag was generally weaker in the endothelial cells lining lymphatic vessels. Staining for the basement membrane component laminin can be used to distinguish lymphatic capillaries and smaller lymphatic collecting vessels from blood vessels.

Antigens↗

[Sclerosis of the intestinal lymphatic vessels].

Fibrosis of the intestinal lymphatic vessels, produced in one case by tuberculosis and, in the other, by appendicitis and peritonitis, caused blockage of the main lymphatic vessels causing, clinically, a protein-losing enteropathy similar to that noted in congenital lymphatic diseases of childhood. In the laboratory, there was noted a fall in serum protein, lipid and cholesterol. A fat absorption test was very abnormal showing a flat curve. During laparotomy, there was discovered on the small intestine, the same layout of lymph vessels, resembling a lace network, as that observed in congenital malformations. Intestinal lymphography showed considerable stasis of the opaque substance and absence of injection of the lymph vessels in the mesentery.

Chylous Ascites↗

Prognostic significance of immunohistochemically detected blood and lymphatic vessel invasion in colorectal carcinoma: its impact on prognosis.

BACKGROUND: The prognostic significance of blood vessel invasion (BVI) and lymphatic vessel invasion (LVI) is unclear. Because of the absence of specific markers for venous and lymphatic vessels, earlier studies could not reliably distinguish between BVI and LVI. METHODS: By immunostaining for podoplanin and CD34 antigen, we retrospectively investigated LVI and BVI in 419 tissue specimens of colorectal carcinoma. We performed univariate and multivariate analysis of the clinicopathologic features, frequency of recurrence, and outcome of patients with or without LVI and BVI. RESULTS: The use of hematoxylin and eosin (H&E) staining to identify BVI and LVI yielded a false positive rate of 9.1% and false negative rate of 12.6%. The incidence of BVI was significantly higher among tumors with LVI than tumors without LVI (P <.001). In logistic multivariate analysis, only LVI (P < .001) was associated with lymph node metastasis and BVI (P = .015) was associated with distant recurrence. Calculating the prognostic relevance, both two invasion types correlated with decreased survival in univariate analysis (both P <.001). In multivariate analysis, BVI (P =.024), lymph node status (P =.003) and tumor stage (P <.001) remained statistically significant factors for survival. CONCLUSIONS: Our results suggest that immunohistologic evaluation of BVI and LVI could be useful in colorectal carcinoma indicating the risk of lymph node metastasis and recurrence, thereby contributing to prognostic evaluation.

Aged↗

Identification of lymphatic vessels in malignant, adenomatous and normal colonic mucosa using the novel immunostain D2-40.

Distribution of lymphatic vessels in normal and neoplastic colon has been previously analyzed with electron microscopic techniques, as reliable antibodies have not been available for selective lymph vessel staining. A novel monoclonal antibody, D2-40, is recently available to differentiate lymphatic vessels from blood vessels. In this study, we analyzed the distribution of lymphatic vessels in normal colon, adenomas with and without superficial stalk invasion and invasive carcinomas without identifiable polypoid precursor lesions. In contrast to previous studies, we found lymphatic vessels in superficially misplaced stalk stroma in adenomas, and closely associated with early invasive epithelial nests in invasive lesions. Lymphatic vessels were identified within the lamina propria of the in situ aspect of in invasive tumors. We conclude that lymphatic vessel structures are seen more superficially in adenomas and invasive carcinomas than previously described. Since intramucosal carcinomas in adenomas do not metastasize, these lymph vessels may be immature or not communicate with deeper lymphatics. Proliferation and distribution of lymphatic vessels may be related to prognosis and early metastasis.

Adenoma↗

Desmoplakin and Plakoglobin--specific markers of lymphatic vessels in the skin?

Monoclonal antibodies against Desmoplakin and Plakoglobin were tested for their suitability as specific markers of lymphatic vessels. The tissue samples were taken from horse skin in an attempt to establish the horse as a model for human lymphatic diseases. To obtain a clear, positive identification of blood and lymphatic vessels, immunohistochemical staining with antibodies against vascular endothelial growth factor receptor 3 (VEGFR-3) and platelet endothelial adhesion molecule (PECAM-1, CD31), was compared with Desmoplakin and Plakoglobin. Because anti-VEGFR-3 is specific for lymphatic vessels in the skin while anti-CD31 stains blood and lymphatic vessels as well, it can be concluded that VEGFR-3(-)/CD31(+) vessels are blood vessels and VEGFR-3(+)/CD31(+) vessels are lymphatic vessels. It was documented on serial sections that Plakoglobin stains both blood and lymphatic vessels. However, Desmoplakin did not stain several positively identified lymphatic vessels. Therefore, Desmoplakin and Plakoglobin antibodies are not specific markers of lymphatic vessels in the skin and the staining pattern is tissues and species dependent.

Animals↗

Lymph and blood vessel architecture in benign and malignant prostatic tissue: lack of lymphangiogenesis in prostate carcinoma assessed with novel lymphatic marker lymphatic vessel endothelial hyaluronan receptor (LYVE-1).

PURPOSE: Due to the lack of specific markers the analysis of lymphatic vessel density (LVD) has been almost impossible in the past. We report the novel specific marker for lymphatic endothelium, lymphatic vessel endothelial hyaluronan receptor (LYVE-1), in prostatic, benign prostatic hyperplasia (BPH) and prostate cancer (PCa) tissue. Normal blood vessels were additionally quantified in BPH and PCa. MATERIALS AND METHODS: LYVE-1 lymphatics (LVD) and CD34 blood vessels were assessed in 20 paraffin sections of BPH and 50 of PCa tissue by immunohistochemistry in a standardized experimental setting. The regions of PCa, periphery of the tumor and nontumorous regions of the PCa specimens, and BPH tissue were evaluated. Double staining was done (LYVE-1/CD34). Acquired data were interrelated and compared to the pathological parameters of the specimens. RESULTS: Double staining revealed numerous CD34 blood vessels but only a few LYVE-1 lymphatic vessels in BPH and PCa sections. Mean LVD +/- SD was distinctly lower (0.55 +/- 0.93) in PCa tissue than in tumor periphery (2.45 +/- 1.93) and nontumorous (3.16 +/- 2.23) tissue (p <0.0001). Maximum LVD was observed in BPH (7.17 +/- 3.61), which differed markedly from nontumorous areas of PCa specimens (p <0.001). In contrast to LVD, significantly more blood vessels were found in PCa (116.00 +/- 39.25) than in BPH (60.30 +/- 19.34) tissue (p <0.001). CONCLUSIONS: LYVE-1 is a specific lymphatic endothelial marker in benign and malignant prostate tissues. It is a useful new marker for the investigation of lymphatics. To our knowledge we report the immunohistochemical visualization and quantification of lymphatic vessels in prostatic tissue for the first time. In contrast to the stimulated angiogenesis of blood vessels in PCa, the destruction of lymphatic vessels occurs rather than lymphangiogenesis.

Antigens, CD34↗

Experimental evaluation of bursting pressure in lymphatic vessels with ultrasonically activated shears.

To evaluate the ability of ultrasonically activated shears (USAS) to occlude lymphatic vessels, we investigated the bursting pressure of lymphatic vessels occluded with USAS and compared with pressures in an artery and a vein. The inguinal lymphatic vessels, testicular arteries, and testicular veins were removed from male pigs. The vessels were occluded by USAS at power level 2. The bursting pressures of the harvested vessels were measured ex vivo. The bursting pressure of the artery (average, 1154 mmHg; range, 1000-1341 mmHg) was significantly greater than that in the vein (average, 747 mmHg; range, 560-973 mmHg; p = 0.0024) or the lymphatic vessel (average, 610 mmHg; range, 491-996 mmHg; p = 0.0003). There was no significant difference of bursting pressure between the vein and lymphatic vessel (p = 0.2226). Although the bursting pressures of the vein and lymphatic vessel were significantly lower than that of the artery, they were much higher than that found in physiologic conditions. USAS is considered to be a powerful tool for lymph node dissection for malignant diseases in open surgery as well as laparoscopic surgery.

Animals↗

Evaluation of the efficacy of the electrosurgical bipolar vessel sealer (LigaSure) devices in sealing lymphatic vessels.

Various sources of ultrasonic and thermal energy have been developed to facilitate blood vessel ligation. However, their efficacy in sealing lymphatics has not been clearly established to date. We hypothesized that the electrosurgical bipolar vessel sealer (EBVS) produces reliable and durable sealing of large lymphatic vessels in a porcine model. Thoracic ducts from 4 adult pigs were explanted and sealed at multiple levels by using 3 different EBVS devices: LigaSure Atlas, XTD, and V. Fifteen seals (5 per group) were analyzed for sealing time and visual quality. Seal burst strength was measured by using a graduated pressure saline injection system. Twelve intact seals also underwent a histologic analysis. The mean overall burst strength of the seals was 271+/-78 mm Hg (127 to 360 mm Hg). The burst pressures in the 3 groups were not statistically different. The overall mean time to achieve a seal was 5.1+/-2.2 seconds (3 to 10 seconds). Seals were achieved significantly faster in the V group (4.1+/-0.6 seconds) compared with the Atlas (6.3+/-2.3 seconds) and XTD (6.4+/-2.6 seconds) groups. Qualitative seal assessment revealed minimal sticking and charring, a favorable degree of seal tissue clarity, and desiccation in the 3 groups. Histologic analysis demonstrated a fusion of lymphovascular channels with a complete obliteration of the lumens. We demonstrated that the use of EBVS results in a fast and effective sealing of large porcine lymphatic vessels. The seals created by all 3 devices burst at markedly supraphysiologic intraluminal pressures. Ongoing randomized human trials may prove the clinical benefits of the routine use of EBVS devices for various tissue dissections.

Animals↗

Periodic acid methenamine silver (pam) staining of the human subcutaneous lymphatic vessel.

The fine structure of the subcutaneous lymphatic vessel taken from the dorsum pedis of the human adult was investigated without and after impregnation by periodic acid methenamine silver (PAM). To check the stainability of another tissue, the renal glomerulus of the rat was also examined in the same way as outlined above. The membrane system of various cells stains positive. No special surface coat of the endothelial cell of the lymphatic vessel was detected. The fine intracellular filament as well as a continuous basal lamina of the endothelial cell were almost PAM-negative, although a part of the basal lamina was occasionally weakly positive. The chemical composition of the basal lamina of the lymphatic vessel is considered to differ from that of the glomerular blood capillary of the kidney, which is PAM-positive. This difference is thought to be due to the smaller polysaccharide content in the former than in the latter.

Adult↗

[Effect of blockage of the afferent lymphatic vessels on the development of popliteal lymph nodes in the rat].

The effect of blockage of the afferent lymphatic vessels on the development of popliteal lymph nodes in the rat was studied. The afferent lymphatic vessels to each popliteal node were surgically interrupted at the lowest edge of the popliteal fossa at 3, 7 or 28 days after birth and the popliteal nodes were obtained from treated animals at 4, 8 or 16 weeks after the operation. At 7 days after birth, each popliteal node was small and weighed 0.2 mg. Its parenchyma consisted of reticular cells and a small number of dispersed lymphoid elements. Four weeks after birth, each node weighed 4-5 mg, and its parenchyma comprised two layers, an outer continuous layer of peripheral cortex containing 40-50 lymph follicles and an inner discontinuous layer of deep cortex made up of 4-5 deep cortical units. At 10-12 weeks after birth, each node weighed about 10 mg and showed full structural development; the peripheral cortex contained 100-130 lymph follicles and the deep cortex consisted of 4-6 well developed units. The popliteal node was drained by 4-6 afferent lymphatic vessels, which opened into the subcapsular sinus of the node. Each lymphatic opening was topographically associated with a respective deep cortical unit, as previously described by Bélisle and Sainte-Marie (1982). In animals treated at 3 or 7 days after birth, the development of the popliteal nodes was considerably inhibited. Four weeks after surgery, each node showed 10-30 lymph follicles in the peripheral cortex and 1-3 small units in the deep cortex. Sixteen weeks after surgery, the node weighed about 4 mg and its cortex exhibited about 50 lymph follicles in the peripheral cortex and only 2 units in the deep cortex. The popliteal nodes of the treated animals generally received 2 afferent lymphatic vessels. In animals treated at 4 weeks after birth, the popliteal nodes showed no gain in weight for following 16 weeks. Four weeks after surgery, each node usually had 4-5 deep cortical units and 50-60 lymph follicles. Thereafter, some units and their overlaying peripheral cortex underwent atrophy, while others persisted. Sixteen weeks after surgery, the popliteal node showed only 2 deep cortical units and 50-60 lymph follicles, and was drained by 2 afferent lymphatic vessels. Surgical interruption of the afferent lymphatic vessels to the popliteal node at the lowest edge of the popliteal fossa did not obliterate all the draining lymphatic vessels , but reduced the number of vessels opening into the node.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Lymphatic vessel density and lymph node metastasis in prostate cancer.

BACKGROUND: Few data are available examining the significance of prostatic lymphatic vessel density (LVD) to lymph node metastasis in patients with prostate cancer. The purpose of this study was to determine the distribution of lymphatic vessels in non-carcinomatous prostate tissue, and investigate the relationship between LVD and lymph node status in prostate cancer. METHODS: LVD, identified by D2-40 immunostaining, was evaluated in non-carcinomatous prostates (n = 7) and prostate cancer (n = 37). The staining pattern of D2-40 was compared with that of another lymphatic vessel marker, vascular endothelial growth factor (VEGF) receptor-3, and a blood vessel endothelial marker, CD34, in adjacent sections. RESULTS: The D2-40 antigen, podoplanin, was expressed exclusively in lymphatic vessels within tumor and normal tissue in all specimens. There was no overlap between cell staining for D2-40 and CD34. Peritumoral LVD and peritumoral lymphatic invasion were significantly associated with lymph node metastasis. VEGF receptor-3 was expressed in a subset of D2-40+ lymphatic vessels. CONCLUSIONS: We have demonstrated that peritumoral lymphatic vessels are likely to serve as major conduits for nodal metastasis in prostate cancer using D2-40 to decorate lymphatic endothelium marker podoplanin. Lack of coexpression of podoplanin and VEGF receptor-3 in some lymphatic vessels suggests the heterogeneity of lymphatic endothelial cells in prostate tissue.

Adenocarcinoma↗

18F-FDG uptake by primary tumor as a predictor of intratumoral lymphatic vessel invasion and lymph node involvement in non-small cell lung cancer: analysis of a multicenter study.

UNLABELLED: Intratumoral lymphatic vessel invasion and lymph node involvement are important factors in the planning of therapeutic strategies, particularly limited surgical resection in patients with non-small cell lung cancer. (18)F-FDG uptake within the primary lesion correlates with aggressiveness on PET studies. The more metabolically active the tumor, the more aggressive are the findings. The aim of this multicenter study was to determine whether (18)F-FDG uptake of the primary tumor is a predictor of intratumoral lymphatic vessel invasion and lymph node metastasis in patients with non-small cell lung cancer. METHODS: One hundred thirty-two patients with lung cancer were studied. All patients underwent a thoracotomy within 4 wk of the (18)F-FDG PET study. A 3-point visual scoring system (low, moderate, or high grade in comparison with mediastinal activity) was used to interpret (18)F-FDG uptake within the primary lesions. The degree of (18)F-FDG uptake in the primary tumor was correlated with the incidence of intratumoral lymphatic vessel invasion and lymph node involvement. Multivariate analysis was performed with logistic multivariate analysis to assess the joint effects and interactions of the variables (age, sex, tumor size, histology, and (18)F-FDG uptake) on intratumoral lymphatic vessel invasion and lymph node involvement. RESULTS: Intratumoral lymphatic vessel invasion and lymph node involvement were found in 7.1% and 5.9%, respectively, of the patients classified in the low-grade group, and in 14.3% and 10.0%, respectively, of the patients classified in the moderate-grade group. In contrast, of the patients classified in the group with high (18)F-FDG uptake, intratumoral lymphatic vessel invasion and lymph node involvement were found in 39.7% and 38.9%, respectively. Multivariate analysis showed that only (18)F-FDG uptake was a significant factor for intratumoral lymphatic vessel invasion and that tumor size and (18)F-FDG uptake were significant factors for lymph node involvement. Of the patients in the high-grade group whose tumors were classified as > or =3 cm in size, lymph node involvement was found in 51.5%. In contrast, of the patients in the low- to moderate-grade group whose tumors were classified as <3 cm in size, lymph node involvement was found in only 9.1% (P < 0.0001). CONCLUSION: Patients with a low to moderate (18)F-FDG uptake in the primary lesion had a significantly lower risk of concurrent intratumoral lymphatic vessel invasion and nodal involvement than did patients with a high (18)F-FDG uptake. In patients with non-small cell lung cancer, (18)F-FDG uptake by the primary tumor is a strong predictor of intratumoral lymphatic vessel invasion and lymph node metastasis.

Adult↗

The distribution and morphology of lymphatic vessels on the peritoneal surface of the adult human diaphragm, as revealed by an ink-absorption method.

Application of india ink to the peritoneal and pleural surfaces of the adult human diaphragm allowed visualization of the distribution and morphology of the lymphatic vessels by light microscopy and scanning electron microscopy. The diaphragms examined had been fixed and stored in 10% formalin. Numerous lymphatic vessels were stained black with india ink, presenting reticular, radial-meshwork, ladder-like and lacy patterns. They were distributed throughout the entire sternocostal part. Analysis by light and scanning electron microscopy of the areas indicated by india ink revealed the presence of primary lymphatic vessels that formed lymphatic lacunae and stomatal openings to the peritoneal cavity. A layer of secondary collecting lymphatic vessels was located cranially with respect to the layer of primary lymphatic vessels. Thus, the peritoneum had at least two layers of lymphatic vessels. These lymphatic vessels were not tubular vessels but resembled flat cisternae, as has been suggested in the case of the mouse diaphragm. The pleura lacked lymphatic stomata and had no such double-layered lymphatic organization. This is the first report that showed distribution and morphology of the lymphatic vessels in the diaphragmatic peritoneum of the formalin-fixed, adult human diaphragm. The method and results in the present study may contribute to morphological analysis of the lymphatic system in the wall of the human body cavity.

Adsorption↗

Immunohistochemical detection of human small lymphatic vessels under normal and pathological conditions using the LYVE-1 antibody.

The spread of tumor cells via lymphatic vessels to the lymph nodes is an important indicator of malignancy. However, previous markers used to identify lymphatic endothelium gave ambiguous results in immunohistochemical analyses with paraffin-embedded tissues. In this study, we attempted to prepare a polyclonal antibody against human lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) for detecting lymphatic vessels using immunohistochemistry. The antibody was raised against a region near the transmembrane anchor of LYVE-1 in New Zealand white rabbits. Immunostainings with anti-LYVE-1 and von Willebrand factor antibodies were performed in various normal and pathological tissues. LYVE-1 expression was confined to the endothelial surface of lymphatic vessels but was not found in the endothelium of blood vessels, which were positive for von Willebrand factor. Our LYVE-1 polyclonal antibody was useful for the identification of small lymphatic vessels in normal human tissues. In addition, the immunostaining enabled us to distinguish lymphatic invasion by malignant tumor cells from blood vessel invasion using paraffin-embedded sections. In conclusion, our polyclonal antibody against the transmembrane anchor of the peptide can be used to detect human lymphatic vessels under various conditions.

Animals↗