Progress in the Mechanism of Lymphangiogenesis and Lymphatic Metastasis of Non-small Cell Lung Cancer / 中国肺癌杂志

Lung cancer ranks the first cancer-related morbidity and mortality in China. Tumor metastasis always predicts the poor prognosis for patients. Moreover, lymphatic metastasis is one of the most significant predictors of poor prognosis in patients with non-small cell lung cancer (NSCLC) and lymphangiogenesis represents the bridge that functionally facilitates tumor lymphatic metastasis. In this review, we first discussed the molecular mechanisms of tumor-associated lymphangiogenesis and the interaction between tumor microenvironment and lymphatic endothelial cells, then, summarized the role of non-coding RNA in regulating tumor-associated lymphangiogenesis in recent frontier studies, with the aim to provide some novel insights on NSCLC-related lymphangiogenesis research, diagnosis and treatment.
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Effect of exosomes derived from human Epstein-Barr virus-positive nasopharyngeal carcinoma cells on lymphangiogenesis and lymph node metastasis / 南方医科大学学报

OBJECTIVE@#To investigate the effect of exosomes derived from Epstein-Barr virus (EBV)-positive nasopharyngeal carcinoma (NPC) cells on lymphangiogenesis and lymph node metastasis of NPC.@*METHODS@#Exosomes from NP69 cells and EBV-positive HK1 (HK1-EBV) cells were obtained by ultracentrifugation and identified by Western blotting and nanoparticle tracking analysis. Dio dye phagocytosis test was performed to observe exosome uptake by lymphatic endothelial cells. Lymphatic endothelial cells were treated with exosomes from nasopharyngeal epithelium (NP69), HK1-EBV, and C666-1 cells or exosome-free supernatant of HK1-EBV and C666-1 cells, and tube formation and migration of the cells were observed. In a nude mouse model of popliteal lymph node metastasis of NPC, the effects of normal saline, NP69 cell-derived exosomes, HK1-EBV cell-derived exosomes, exosome-free supernatant of HK1-EBV cells, and HK1-EBV exosome-free supernatant protein on lymphangiogenesis and lymph node metastasis of the tumor were observed.@*RESULTS@#The exosomes obtained by ultracentrifugation contained abundant exosome-specific proteins and showed a normal size range. The exosomes from NPC cells and NP69 cells could be taken up by lymphatic endothelial cells. Compared with the blank control and exosomes form NP69 cells, exosomes derived from HK1-EBV and C666-1 cells significantly promoted tube formation and migration of lymphatic endothelial cells (@*CONCLUSIONS@#Exosomes from EBV-positive NPC cells can significantly promote lymphangiogenesis and lymph node metastasis of NPC.

Síndromes MonoMAC e Emberger em paciente com mutação no gene GATA2 / MonoMAC and Emberger syndromes in a patient with GATA2 mutation

As mutações que ocorrem no gene GATA2 podem ocasionar um amplo espectro de doenças genéticas. Os pacientes podem ter anormalidades na hematopoiese, na linfangiogenesis e na resposta imunológica. Os fenótipos incluem algumas síndromes caracterizadas por monocitopenia e infecção por micobactéria (síndrome MonoMac), síndrome mielodisplásica familiar, leucemia mieloide crônica ou aguda, síndrome de Emberger (linfedema primário), e mais raramente neutropenia, anemia aplástica e deficiência isolada de células NK. A idade da apresentação clínica pode variar desde a infância até a idade adulta. A deficiência autossômica dominante de GATA2 pode permanecer clinicamente silenciosa por décadas, ou mesmo durante toda a vida. Descrevemos o caso de uma jovem brasileira que apresentou a maioria dos problemas ligados à mutação no gene GATA2, observando-se as duas síndromes: MonoMAC e Emberger.

GATA2 mutations may cause a wide spectrum of genetic disorders. Patients may have several abnormalities in hematopoiesis, lymphangiogenesis and immune response. The phenotypes include monocytopenia and mycobacterial infection (MonoMAC) syndrome, familial myelodysplastic syndrome (MDS), chronic or acute myeloid leukemia (CML or AML), Emberger syndrome and, more rarely, neutropenia, aplastic anemia and isolated NKcell deficiency. Age at clinical onset ranges from early childhood to late adulthood. Autosomal dominant GATA2 deficiency may remain clinically silent for decades or even for life. We report a case of a Brazilian young patient who had most of the problems related to GATA2 mutation as well as MonoMAC and Emberger syndromes.

Increased expression of vascular endothelial growth factor-C and vascular endothelial growth factor receptor-3 after pilocarpine-induced status epilepticus in mice

Vascular endothelial growth factor (VEGF)-C and its receptor, vascular endothelial growth factor receptor (VEGFR)-3, are responsible for lymphangiogenesis in both embryos and adults. In epilepsy, the expression of VEGF-C and VEGFR-3 was significantly upregulated in the human brains affected with temporal lobe epilepsy. Moreover, pharmacologic inhibition of VEGF receptors after acute seizures could suppress the generation of spontaneous recurrent seizures, suggesting a critical role of VEGF-related signaling in epilepsy. Therefore, in the present study, the spatiotemporal expression of VEGF-C and VEGFR-3 against pilocarpine-induced status epilepticus (SE) was investigated in C57BL/6N mice using immunohistochemistry. At 1 day after SE, hippocampal astrocytes and microglia were activated. Pyramidal neuronal death was observed at 4 days after SE. In the subpyramidal zone, VEGF-C expression gradually increased and peaked at 7 days after SE, while VEGFR-3 was significantly upregulated at 4 days after SE and began to decrease at 7 days after SE. Most VEGF-C/VEGFR-3-expressing cells were pyramidal neurons, but VEGF-C was also observed in some astrocytes in sham-manipulated animals. However, at 4 days and 7 days after SE, both VEGFR-3 and VEGF-C immunoreactivities were observed mainly in astrocytes and in some microglia of the stratum radiatum and lacunosum-moleculare of the hippocampus, respectively. These data indicate that VEGF-C and VEGFR-3 can be upregulated in hippocampal astrocytes and microglia after pilocarpine-induced SE, providing basic information about VEGF-C and VEGFR-3 expression patterns following acute seizures.

Association Between c-Met and Lymphangiogenic Factors in Patients With Colorectal Cancer

PURPOSE: Animal models show a strong relationship between lymphangiogenesis and lymph node metastasis. However, the clinical significance of lymphangiogenesis in patients with colorectal cancer (CRC) remains uncertain. This study aimed to evaluate the association between c-Met and lymphangiogenic factors and to elucidate the prognostic significance of c-Met in patients with CRC. METHODS: A total of 379 tissue samples were obtained from surgically resected specimens from patients with CRC at Soonchunhyang University Cheonan Hospital between January 2002 and December 2010. The expressions of c-Met, vascular endothelial growth factor (VEGF)-C, VEGF-D, VEGF receptor (VEGFR)-3, and podoplanin were examined using immunohistochemistry. The expression of c-Met and clinical factors were analyzed. RESULTS: Of the 379 tissues, 301 (79.4%) had c-Met expression. High expression of c-Met in tumor cells was significantly associated with high expression of VEGF-C (P < 0.001) and VEGFR-3 (P = 0.001). However, no statistically significant association with podoplanin (P = 0.587) or VEGF-D (P = 0.096) was found. Of the 103 evaluable patients, expression of c-Met in tumor cells was significantly associated with advanced clinical stage (P = 0.020), positive lymph node status (P = 0.038), and high expression of VEGF-C (P = 0.020). However, no statistically significant association with podoplanin (P = 0.518), VEGFR-3 (P = 0.085), VEGF-D (P = 0.203), or overall survival (P = 0.360) was found. CONCLUSION: Our results provide indirect evidence for an association and possible regulatory link of c-Met with the lymphangiogenic markers, but c-Met expression in patients with CRC is not a prognostic indicator for overall survival.

Changes of lymphatic vessel density in lung adenocarcinoma in situ, minimally invasive adenocarcinoma, and invasive adenocarcinoma and the regulatory factors / 南方医科大学学报

OBJECTIVE@#To analyze the changes in tumor lymphatic vessel density (LVD) in patients with lung adenocarcinoma in situ (AIS), minimally invasive adenocarcinoma (MIA), and invasive adenocarcinoma (IA) and explore the regulatory factors of LVD.@*METHODS@#Complete clinicopathological data were collected form a total of 301 patients with lung adenocarcinoma, including 28 (9.3%) with AIS, 86 (28.6%) with MIA, and 187 (62.1%) with IA. The LVD of all the adenocarcinomas were calculated after D2-40 immunohistochemical staining, and MT1-MMP and VEGF-C expression levels were also evaluated. The differences in LVD among the groups and the correlations of tumor LVD with the expressions of MT1-MMP and VEGF-C and the clinicopathological factors were analyzed.@*RESULTS@#The LVD differed significantly among AIS, MIA, and IA groups (= 0.000). The LVDs was significantly correlated with the level of VEGF-C protein expression (=0.917, =0.009), tumor size (= 0.686, =0.017), lymph node metastasis (=0.739, =0.000), and clinical stage (=0.874, =0.012) of the patients.@*CONCLUSIONS@#Tumor lymphangiogenesis plays an important role in lung adenocarcinoma progression, and VEGF-C may promote this process.

Emerging Roles of Lymphatic Vasculature in Immunity

The lymphatic vasculature has been regarded as a passive conduit for interstitial fluid and responsible for the absorption of macromolecules such as proteins or lipids and transport of nutrients from food. However, emerging data show that the lymphatic vasculature system plays an important role in immune modulation. One of its major roles is to coordinate antigen transport and immune-cell trafficking from peripheral tissues to secondary lymphoid organs, lymph nodes. This perspective was recently updated with the notion that the interaction between lymphatic endothelial cells and leukocytes controls the immune-cell migration and immune responses by regulating lymphatic flow and various secreted molecules such as chemokines and cytokines. In this review, we introduce the lymphatic vasculature networks and genetic transgenic models for research on the lymphatic vasculature system. Next, we discuss the contribution of lymphatic endothelial cells to the control of immune-cell trafficking and to maintenance of peripheral tolerance. Finally, the physiological roles and features of the lymphatic vasculature system are further discussed regarding inflammation-induced lymphangiogenesis in a pathological condition, especially in mucosal tissues such as the gastrointestinal tract and respiratory tract.

Lymphangiogenesis and angiogenesis in oral cavity and lower lip squamous cell carcinoma / Linfangiogênese e angiogênese em carcinomas de células escamosas de lábio inferior e da cavidade oral

ABSTRACT INTRODUCTION: Tumors of the lip and oral cavity differ in various aspects; therefore a clarification of the distinctions among these sites may help to better understand the biologic behavior of neoplasms occurring in these locations. OBJECTIVE: Considering that angiogenesis and lymphangiogenesis are two major elements that can influence various aspects of tumor biology, we aimed to compare these factors between squamous cell carcinoma of the lower lip and oral cavity. METHODS: A total of 84 primary squamous cell carcinomas including 45 oral and 39 lower lip tumors were selected and immunohistochemically stained with monoclonal antibody against D2-40 and CD105. Mean microvessel density was assessed in tumoral tissue, while lymphatic vessel density was calculated in both neoplastic tissue and invasion front. Data were statistically analyzed using t-test and p-values of <0.05 were considered significant. RESULTS: We found a mean microvessel density ± standard deviation of 31.94 ± 18.9 in oral cavity and 27.54 ± 20.8 in lower lip squamous cell carcinomas, with no significant difference (p = 0.32). Mean lymphatic vessel density ± standard deviation was 13.05 ± 8.2 and 16.57 ± 10.79 in of oral cavity and lower lip neoplastic tissue, respectively. The corresponding values were 9.94 ± 5.59 and 12.50 ± 7.8 in the invasive front. Significant differences were not observed in either of the lymphatic vessel density variables between the two sites. CONCLUSION: According to our results, it seems that the search for additional factors other than those related to the vasculature should continue, to help clarify the differences in biologic behavior between lower lip and oral cavity squamous cell carcinomas.

Resumo Introdução: Os tumores de lábio e da cavidade oral diferem em vários aspectos; portanto, o conhecimento das diferenças entre eles pode ajudar na melhor compreensão do comportamento biológico das neoplasias que ocorrem nesses locais. Objetivo: Considerando que a angiogênese e a linfangiogênese são dois elementos importantes que podem influenciar diversos aspectos da biologia dos tumores, objetivamos comparar esses fatores entre o carcinoma de células escamosas (CCE) de lábio inferior e da cavidade oral. Método: No total, foram selecionados 84 CCEs primários (45 tumores da cavidade oral e 39 tumores de lábio). Esses tumores foram corados por processo imunohistoquímico com anticorpo monoclonal anti-D2-40 e CD105. Avaliamos a densidade média de microvasos (DMV) no tecido tumoral, enquanto que a densidade vascular linfática (DVL) foi calculada tanto no tecido neoplásico como no front de invasão. Os dados foram estatisticamente analisados com o uso do teste t e valores de p < 0,05 foram considerados significantes. Resultados: Chegamos a uma média para DMV ± DP de 31,94 ± 18,9 para CCEs na cavidade oral e de 27,54 ± 20,8 no lábio inferior, sem diferença significante (p = 0,32). As médias para DVL ± DP foram de 13,05 ± 8,2 e 16,57 ± 10,79 no tecido neoplásico da cavidade oral e lábio inferior, respectivamente. Os valores correspondentes foram 9,94 ± 5,59 e 12,50 ± 7,8 no front invasivo. Não foram observadas diferenças significantes nas duas variáveis DVL entre os dois locais. Conclusão: De acordo com os nossos resultados, a pesquisa por fatores adicionais, além daqueles relacionados à vasculatura, deve ter continuidade, para auxiliar no esclarecimento das diferenças do comportamento biológico entre CCEs no lábio inferior e na cavidade oral.

Expressão de VEGF-C, VEGF-D, mensuração da densidade linfática e da proliferação endotelial linfática em neoplasias de glândulas salivar / Expression of VEGF-C, VEGF-D density measurement of lymphatic and lymphatic endothelial proliferation in salivary gland neoplasms

As neoplasias de glândulas salivares exibem uma grande diversidade morfológica e comportamentos biológicos variados o que suscita o interesse na pesquisa destas lesões. A disseminação das células tumorais é um passo inicial para a progressão de neoplasias malignas e, dentro deste contexto, os vasos linfáticos neoformados são considerados essenciais para que ocorra essa disseminação. O papel do VEGF (fator de crescimento endotelial vascular) na formação dos vasos é fato conhecido mas, pouco se sabe a respeito de sua participação em tumores de glândula salivar. Desta forma, o objetivo deste estudo foi avaliar a expressão do VEGF-C e VEGF-D, a densidade linfática tumoral (D2-40) e a proliferação endotelial linfática (dupla marcação D2-40/Ki-67) em uma série de neoplasias de glândulas salivares. A amostra foi composta por 20 adenomas pleomórficos, 20 carcinomas adenóides císticos, 20 carcinomas mucoepidermóides e 10 casos de tecido glandular salivar com características de normalidade para efeito comparativo. Todos os casos estudados exibiram expressão positiva para VEGF-C em região peritumoral e intratumoral, não sendo encontrada diferenças de imunoexpressão entre os grupos. No entanto, o grupo dos carcinomas adenóides císticos demonstrou diferença significativa da imunoexpressão do VEGF-C segundo o padrão cribriforme e sólido (p = 0,004). A maioria dos casos constantes do presente estudo, apresentou fraca marcação para VEGF-D em região peritumoral e intratumoral...

Salivary gland neoplasms exhibit a great morphological diversity and varied biological behavior which raises the interest in the study of these lesions. The spread of tumor cells is an early step in the progression of malignancies and the neoformed lymphatic vessels are considered essential in tumor dissemination. Vascular endotelial growth fator (VEGF) is a family of proteins involved in angiogenesis e lymphangiogenesis. However, in salivar tumors we have limited information on the expression. The aim of this study was to assess the expression of VEGF-C and VEGF-D, lymphatic vessel density (single-staining D2-40) and lymphatic endothelial proliferation (double labeling D2-40/Ki-67) in a series of salivary glands neoplasms. We selected 20 cases of pleomorphic adenoma, 20 of mucoepidermoide carcinoma, 20 of adenoid cystic carcinoma and 10 tissue sample of normal salivary gland. All cases studied showed positive expression of VEGF-C in intratumoral and peritumoral region, no differences in immunoreactivity was found between the groups. However, the group of adenoid cystic carcinoma showed a significant difference in immunoreactivity of VEGF-C by the cribriform and solid pattern (p = 0.004). Most of the cases included in this study showed weak immunoreactivity for VEGF-D in intratumoral and peritumoral region. In the assessment of lymphatic endotelial density peritumoral, intratumoral and total, the groups showed an increasing gradient, with lower values for the group of pleomorphic adenomas followed by mucoepidermoid carcinoma and adenoid cystic carcinoma. Lymphatic endothelial cell density was higher in malignant than benign tumors. No correlation was observed between the immunoreactivity of VEGF-C and VEGF-D in relation to tumor lymphatic density and lymphatic endothelial proliferation.

Promoting inflammatory lymphangiogenesis by vascular endothelial growth factor-C (VEGF-C) aggravated intestinal inflammation in mice with experimental acute colitis

Angiogenesis and lymphangiogenesis are thought to play a role in the pathogenesis of inflammatory bowel diseases (IBD). However, it is not understood if inflammatory lymphangiogenesis is a pathological consequence or a productive attempt to resolve the inflammation. This study investigated the effect of lymphangiogenesis on intestinal inflammation by overexpressing a lymphangiogenesis factor, vascular endothelial growth factor-C (VEGF-C), in a mouse model of acute colitis. Forty eight-week-old female C57BL/6 mice were treated with recombinant adenovirus overexpressing VEGF-C or with recombinant VEGF-C156S protein. Acute colitis was then established by exposing the mice to 5% dextran sodium sulfate (DSS) for 7 days. Mice were evaluated for disease activity index (DAI), colonic inflammatory changes, colon edema, microvessel density, lymphatic vessel density (LVD), and VEGFR-3mRNA expression in colon tissue. When acute colitis was induced in mice overexpressing VEGF-C, there was a significant increase in colonic epithelial damage, inflammatory edema, microvessel density, and neutrophil infiltration compared to control mice. These mice also exhibited increased lymphatic vessel density (73.0±3.9 vs 38.2±1.9, P<0.001) and lymphatic vessel size (1974.6±104.3 vs 1639.0±91.5, P<0.001) compared to control mice. Additionally, the expression of VEGFR-3 mRNA was significantly upregulated in VEGF-C156S mice compared to DSS-treated mice after induction of colitis (42.0±1.4 vs 3.5±0.4, P<0.001). Stimulation of lymphangiogenesis by VEGF-C during acute colitis promoted inflammatory lymphangiogenesis in the colon and aggravated intestinal inflammation. Inflammatory lymphangiogenesis may have pleiotropic effects at different stages of IBD.

Lymphangiogenesis in Breast Cancer Correlates with Matrix Stiffness on Shear-Wave Elastography

PURPOSE: To correlate tumor stiffness and lymphangiogenesis in breast cancer and to find its clinical implications. MATERIALS AND METHODS: A total of 140 breast cancer patients were evaluated. Tumor stiffness was quantitatively measured by shear-wave elastography in preoperative ultrasound examination, calculated as mean elasticity value (kPa). Slides of resected breast cancer specimens were reviewed for most fibrotic area associated with tumor. D2-40 immunohistochemical staining was applied for fibrotic areas to detect the lymphatic spaces. Microlymphatic density, tumor stiffness, and clinicopathologic data were analyzed. RESULTS: Higher elasticity value was associated with invasive size of tumor, microlymphatic density, histologic grade 3, absence of extensive intraductal component, presence of axillary lymph node metastasis, and Ki-67 labeling index (LI) in univariate regression analysis, and associated with Ki-67 LI and axillary lymph node metastasis in multivariate regression analysis. Microlymphatic density was associated histologic grade 3, mean elasticity value, and Ki-67 LI in univariate regression analysis. In multivariate regression analysis, microlymphatic density was correlated with mean elasticity value. CONCLUSION: In breast cancer, tumor stiffness correlates with lymphangiogenesis and poor prognostic factors.

Mechanisms of Lymph Node Metastasis in Head and Neck Cancer / 대한이비인후과학회지

Lymph node metastasis (LNM) is an important prognostic factor in head and neck cancer (HNC), which leads to recurrence and poor outcome. Despite the advances in multimodality treatment protocols, overall survival in patients with LNM remains limited, thus calling for the need of a more thorough understanding of the biology in metastatic process. In the past, LNM had been suspected to rely mostly on passive mechanistic impulse from primary tumor. However, recent discovery of new lymphatic markers, regulating growth factors, and cognate receptors, has elucidated the active biological regulation during metastatic cascades, which primarily involves primary tumor lymphangiogenesis, pre-metastatic niche formation, and sentinel LNM. In this review, we discuss recent literatures on the mechanisms of LNM in HNC that is expected to allow a better understanding of the disease as well as suggesting a potential clinical implication.

A linfangiogênese em enxertos corneanos humanos que evoluem para retransplante / Lymphangiogenesis in human corneal grafts that has evolved to re-transplantation

RESUMO Objetivo: Estudar botões corneanos humanos com linfangiogênese através do exame histopatológico, juntamente com os enxertos de seus transplantes anteriores e posteriores, avaliando os intervalos de tempo para sucessivas cirurgias. Métodos: Estudo descritivo, observacional, longitudinal de botões corneanos humanos com linfangiogênese, juntamente com seus transplantes anteriores e posteriores. Os tecidos foram provenientes de ceratoplastia penetrante no período compreendido entre os anos 2006 e 2013. Após revisão de prontuários em que foram obtidas principalmente as datas das cirurgias, construímos uma tábua de sobrevivência a partir da qual os intervalos de tempo para retransplante foram calculados. Resultados: Entre 89 casos de linfangiogênese corneana, foram incluídos apenas aqueles 22 que possuíam registros no prontuário de transplantes anteriores ou posteriores. Nos casos que apresentavam como provável etiologia do retransplante a linfangiogênese, isolada ou associada à hemangiogênese (grupos pré-linfangiogênese/linfangiogênese e interlinfangiogênese), foram encontrados intervalos de tempo para retransplante menores (7 e 3 meses, respectivamente) que aquele encontrado no grupo linfangiogênese/pós-linfangiogênese que apresentava outras etiologias prováveis para os retransplantes (11,31 meses). Casos que apresentavam como etiologia provável do retransplante a linfangiogênese isolada apresentaram um intervalo para retransplante (3 meses) ainda menor que aquele encontrado nos casos em que a etiologia provável era a linfangiogênese associada à hemangiogênese (7,80 meses). Conclusão: Linfangiogênese, isolada ou associada à hemangiogênese, foi encontrada nos enxertos corneanos humanos estudados que evoluíram para retransplante em pequenos intervalos de tempo. Esse achado nos leva a sugerir um possível papel para os vasos linfáticos na redução do tempo de sobrevida dos enxertos corneanos humanos.

ABSTRACT Objective: To study human corneal buttons with lymphangiogenesis through histopathological examination, together with the grafts of their preceding and subsequent transplantations, evaluating the time intervals for successive surgeries Methods: A descriptive, observational and longitudinal study of human corneal buttons that have lymphatic vessels, together with its preceding and subsequent transplants. Tissues were obtained from penetrating keratoplasty in the period between the years 2006 and 2013. After a medical records review in which information on the dates of the surgeries were mainly obtained, we built a survival table from which the time intervals for retransplantation were calculated. Results: Among 89 cases of corneal lymphangiogenesis, we included only those 22, which had previous or subsequent transplantations records in medical records. In cases where the probable regrafting etiology were lymphangiogenesis, alone or combined with hemangiogenesis (pre-lymphangiogenesis/lymphangiogenesis and interlymphangiogenesis groups), time intervals for retransplantation were found to be minor (7 and 3 months, respectively) than that found in lymphangiogenesis/post-lymphangiogenesis group that had other probable etiologies for retransplantations (11.31 months). Cases that had isolated lymphangiogenesis as probable etiology of retransplantation showed an interval time for retransplantation (3 months) lower than that found in cases in which the probable etiology was lymphangiogenesis associated with hemangiogenesis (7.80 months). Conclusion: Lymphangiogenesis, alone or combined with hemangiogenesis, was found in human corneal grafts studied that have evolved to regraft in small time intervals. This finding leads us to suggest a possible role for the lymphatic vessels in reducing the human corneal grafts survival time.

The main findings in histopathological examination of human corneal buttons with lymphangiogenesis / Principais achados do exame histopatológico de botões corneanos humanos com linfangiogênese

Objective: To detect the presence of lymphatic vessels in the cornea through histopathological examination, trying to identify findings that are most commonly found with the presence of these vessels in this tissue. Methods: Retrospective descriptive study of human corneal buttons with lymphangiogenesis. Tissues were obtained from penetrating keratoplasty in the period between the years 2006 and 2013. A medical record review was conducted looking for information about sex, age and graft etiology. Results: 89 corneal buttons were included, out of which 37 were from female patients and 52 were from male patients. The average age was 47.70 ± 23.95 years (mean ± SD). Lymphangiogenesis was found mainly associated with hemangiogenesis. However, isolated lymphangiogenesis was observed in 28 (31.46%) patients. In 18 (20.22%) cases were found an amount of lymphatic vessels approximately four times higher than that found in most part of the sample. A lot of cases were found in inflammatory conditions such as infection and perforation. Near the lymphangiogenesis, we found many cases of anterior synechia and myofibroblasts. In 17 (16.35%) cases, no change was observed in the vicinity of corneal lymphatic vessels. Conclusions: We demonstrated through a histopathological examination, that findings admittedly associated with lymphangiogenesis like inflamamatory processes, are also frequently found in cases of human corneas that have lymphatic vessels. However, other findings such as lymphangiogenesis without the presence of angiogenesis, the presence of a greater amount of vessels in some cases and lymphangiogenesis without changes in its proximity remain in need of a better understanding.

Objetivo: Detectar a presença de vasos linfáticos na córnea através do exame histopatológico, buscando identificar os achados que são encontrados com maior frequência junto à presença desses vasos nesse tecido. Métodos: Estudo retrospectivo descritivo de botões corneanos humanos com linfangiogênese. Os tecidos foram provenientes de ceratoplastia penetrante no período compreendido entre os anos de 2006 e 2013. Foi realizada revisão de prontuários em busca de informações sobre sexo, idade e etiologia do transplante. Resultados: Foram incluídos 89 botões corneanos, sendo 37 de pacientes do sexo feminino e 52 do sexo masculino. A média das idades foi de 47,70 ± 23,95 anos (média ± DP). Linfangiogênese foi encontrada principalmente associada à hemangiogênese. Linfangiogênese isolada, no entanto, foi observada em 28 (31,46%) casos. Em 18 (20,22%) casos foram encontrados uma quantidade de vasos linfáticos cerca de quatro vezes maior que aquela encontrada na maioria das amostras. Em um grande número de casos foram encontrados condições inflamatórias como infecção e perfuração. Nas proximidades da linfangiogênese, encontramos muitos casos de sinéquia anterior e miofibroblastos. Em 17 (16,35%) casos nenhuma alteração foi evidenciada nas proximidades dos vasos linfáticos corneanos. Conclusão: Demonstramos, através do exame histopatológico, que achados reconhecidamente associados à linfangiogênese, como os processos inflamatórios, são encontrados também com frequência em casos de córneas humanas que possuem vasos linfáticos. Porém, outros achados evidenciados, como a linfangiogênese desacompanhada de angiogênese, a presença de uma maior quantidade de vasos em alguns casos e a linfangiogênese sem alterações em sua proximidade, permanecem necessitando de uma melhor compreensão.

The Effect of Subconjunctival Bevacizumab Injection before Conjunctival Autograft for Pterygium

PURPOSE: To evaluate the effect of subconjunctival bevacizumab injection before conjunctival autograft for pterygium. METHODS: Twenty-five eyes (25 patients) with pterygium received a subconjunctival injection of 2.5 mg (0.1 mL) bevacizumab 1-2 weeks prior to conjunctival autograft surgery. The control group (25 eyes of 25 patients) received the same operation. Two weeks, 1 month and every month after the surgery, the vascularization of surgical site, the recurrence rate and the effect of wound healing were analyzed. RESULTS: The bevacizumab group showed a decreased conjunctival vascularity grade compared with the control group based on light microscopy. The bevacizumab group also showed lower vascular epithelial growth factor (VEGF) compared with the control group using immunohistochemical analysis and western blot. There was no recurrence in both groups, but, persistent autograft edema was observed at 8 weeks postoperatively in the bevacizumab group. CONCLUSIONS: Although preoperative injection of bevacizumab effectively reduced vascularity and VEGF concentration of pterygium tissue, prolonged autograft edema was observed. Based on these results, bevacizumab inhibits lymphangiogenesis as well as angiogenesis. Therefore, delayed wound healing should be considered when subconjunctival bevacizumab injection is administered before pterygium surgery.

Peri-articular lymphatic system and "Bi" theory of Chinese medicine in the pathogenesis and treatment of arthritis / 中国结合医学杂志

Rheumatoid arthritis (RA) and osteoarthritis (OA) are the two most common joint diseases, and they have characterization of synovial inflammation and cartilage destruction, associated with the accumulation of numerous catabolic mediators and inflammatory cells in the synovial space and surrounding soft tissues. How these factors are cleared and if the "clearance" process contributes to pathogenesis of arthritis are not known. Recently, we found the existence of the peri-articular lymphatic system in mouse joints. The blockade of lymphangiogenesis and lymphatic draining function accelerates while stimulation of lymphatic function attenuates the severity of joint tissue lesions in mouse models of RA and OA. More importantly, we noticed the similarity between the dysfunction of lymphatic drainage in arthritic joints and "Bi" theory of Chinese medicine (CM), and demonstrated that several Bi disease-treated herbal drugs directly affect the function of lymphatic endothelial cells. Here we review the advances about the interactions between joint inflammation and changes in the peri-articular lymphatic system and discuss our view of linking "Bi" theory of CM to lymphatic dysfunction in arthritis.

Tight junction protein claudin 4 in gastric carcinoma and its relation to lymphangiogenic activity

Background and study aims: Gastric cancer is the second most common cause of cancer-related death worldwide. Claudins are a family of tight junction proteins that are biologically relevant in many cancer progression steps. This study aimed to investigate the expression of the intestinal claudin [claudin 4] in gastric carcinoma and to evaluate its relation to the different clinicopathologic prognostic parameters, especially lymphangiogenesis [production of new lymphatic vessels, measured by lymphovascular density [LVD]] and lymphovascular invasion [LVI]

Patients and methods: Fifty-five gastric carcinoma specimens were immunohistochemically stained for claudin 4 and D2-40 [for detection of lymphatic vessel endothelium]

Results: High expression of claudin 4 was detected in 26 of 55 [47.3%] cases. Low expression of claudin 4 was related to poorly differentiated type [p = 0.001], non-intestinal [diffuse] type [p = 0.001], deeper tumour invasion [p < 0.001], lymph node metastasis [p = 0.001], and higher stage [p = 0.001]. In addition, higher LVD was related to poorly differentiated types [p = 0.001], non-intestinal type [p = 0.001], lymph node metastasis [p = 0.015], and higher tumour, node, metastasis [TNM] stage [p = 0.001]. LVI was related to lymph node metastasis [p = 0.025], higher TNM stage [p = 0.001], and LVD [p = 0.001]. Claudin 4 significantly correlated with both LVD [p = 0.009] and LVI [p = 0.009]

Conclusions: High expression of claudin 4 was associated with the more differentiated intestinal-type gastric carcinoma and lost in poorly differentiated diffuse type. So, claudin 4 may be used as one of the differentiating markers between the two major types of gastric carcinoma [intestinal vs. diffuse]. LVD and LVI were related to higher incidence of lymph node metastasis and therefore could be used as predictive markers for lymph node metastasis in limited specimens during early gastric carcinoma to determine the need for more invasive surgery. Low expression of claudin 4 was related to lymphangiogenesis. This may shed light on the relation of tight junction protein expression and lymphangiogenesis

Expression of lymphatic markers and lymphatic growth factors in psoriasis before and after anti-TNF treatment

BACKGROUND: Angiogenesis is an early stage of psoriatic lesion development, but less is known about lymphagiogenesis and its role in the development of psoriasis. OBJECTIVE: To examine the expression of specific lymphatic markers and lymphatic growth factors in untreated psoriatic skin, in the unaffected skin of patients and skin of healthy volunteers, as well as their alteration after treatment with an anti-TNF agent. METHODS: Immunohistochemistry for the lymphatic markers D2-40 and LYVE-1, in addition to the VEGF-C and VEGF-D growth factors, was performed in the skin biopsies of psoriatic lesions and adjacent non-psoriatic skin of 19 patients before and after treatment with etanercept, as well as in the skin biopsies of 10 healthy volunteers. RESULTS: The expressions of D2-40, VEGF-C and VEGF-D on lymphatic vessels underwent statistically significant increases in untreated psoriatic skin compared with non-lesional skin, in contrast to LYVE-1, which did not involve significant increase in expression in psoriatic skin. VEGF-C expression on lymphatic vessels diminished after treatment with etanercept. Moreover VEGF-C and VEGF-D staining on fibroblasts presented with higher expression in lesional skin than in non-lesional adjacent skin. CONCLUSION: Remodeling of lymphatic vessels possibly occurs during psoriatic lesion development, parallel to blood vessel formation. The exact role of this alteration is not yet clear and more studies are necessary to confirm these results. .

FASN expression, angiogenesis and lymphangiogenesis in central and peripheral giant cell lesions

Central giant cell lesion (CGCL) and peripheral giant cell lesion (PGCL) are non-neoplastic proliferative processes of the jaws. PGCL is a reactive process induced by irritant local factors and CGCL is an intra-osseous lesion of unknown etiology. Both lesions exhibit similar histologic features showing abundant mononuclear cells, admixed with a large number of multinucleated giant cells and a rich vascularized stroma with extravasated erythrocytes, hemosiderin deposition, and blood-filled pools. Recent studies have linked fatty acid synthase (FASN) with angiogenesis. Objective: To evaluate angiogenesis and lymphangiogenesis and their relationship with FASN expression in CGCL and PGCL. Material and Methods: Thirteen CGCL and 14 PGCL of the jaws were selected for immunoexpression of FASN; CD34 and CD105 (to assess blood microvessel density [MVD] and microvessel area [MVA]); and D2-40 (to assess lymphatic MVD and MVA). Results: Within PGCL and CGCL, MVD-CD34 was signifcantly higher than MVD-CD10S, followed by MVD-D2-40. Moreover, a signifcantly higher number of FASN-positive multinucleated giant cells than mononuclear cells were observed. Between PGCL and CGCL, only MVD-CD34 and all MVA were signifcantly higher in PGCL. Positive correlation between MVA-CD10S with FASNpositive mononuclear cells in both lesions was observed. Conclusions: Our results show both lesions exhibiting similar levels of FASN expression and neoangiogenesis, suggesting constitutive processes that regulate tissue maintenance. .

(Lymph)angiogenic influences on hematopoietic cells in acute myeloid leukemia

The purpose of this review is to provide an overview of the effect of (lymph)angiogenic cytokines on hematopoietic cells involved in acute myeloid leukemia (AML). Like angiogenesis, lymphangiogenesis occurs in pathophysiological conditions but not in healthy adults. AML is closely associated with the vasculature system, and the interplay between lymphangiogenic cytokines maintains leukemic blast survival in the bone marrow (BM). Once AML is induced, proangiogenic cytokines function as angiogenic or lymphangiogenic factors and affect hematopoietic cells, including BM-derived immune cells. Simultaneously, the representative cytokines, VEGFs and their receptors are expressed on AML blasts in vascular and osteoblast niches in both the BM and the peripheral circulation. After exposure to (lymph)angiogenic cytokines in leukemogenesis and infiltration, immune cell phenotypes and functions are affected. These dynamic behaviors in the BM reflect the clinical features of AML. In this review, we note the importance of lymphangiogenic factors and their receptors in hematopoietic cells in AML. Understanding the functional characterization of (lymph)angiogenic factors in the BM niche in AML will also be helpful in interrupting the engraftment of leukemic stem cells and for enhancing immune cell function by modulating the tumor microenvironment.