Lymphatic Research Articles

Congenital chylothorax managed with octreotide and skimmed milk: a case report

Pleural effusion occurs when a large amount of free fluid accumulates in the pleural space. A chylothorax is caused by chyle-containing lymphatic fluid draining into the pleural cavity. This is the most common type of pleural effusion observed during the neonatal period. It causes a number of respiratory and nutritional issues and has a significant mortality rate. It is usually treated by intercostal chest tube drainage and total parenteral nutrition. Octreotide is a relatively newer strategy in the management. Here we describe the case of a 16 days old male neonate who presented to us with sudden onset severe respiratory distress requiring invasive ventilation. Radiology suggested pleural effusion and so intercostal chest tube was inserted, which revealed Chylous effusion. The patient was successfully managed with dietary therapy including skimmed milk and octreotide infusion.

The Lymphatic System in Kidney Disease.

The high-capacity vessels of the lymphatic system drain extravasated fluid and macromolecules from nearly every part of the body. However, far from merely a passive conduit for fluid removal, the lymphatic system also plays a critical and active role in immune surveillance and immune response modulation through the presentation of fluid, macromolecules, and trafficking immune cells to surveillance cells in regional draining lymph nodes before their return to the systemic circulation. The potential effect of this system in numerous disease states both within and outside of the kidney is increasingly being explored for their therapeutic potential. In the kidneys, the lymphatics play a critical role in both fluid and macromolecule removal to maintain oncotic and hydrostatic pressure gradients for normal kidney function, as well as in shaping kidney immunity, and potentially in balancing physiological pathways that promote healthy organ maintenance and responses to injury. In many states of kidney disease, including AKI, the demand on the preexisting lymphatic network increases for clearance of injury-related tissue edema and inflammatory infiltrates. Lymphangiogenesis, stimulated by macrophages, injured resident cells, and other drivers in kidney tissue, is highly prevalent in settings of AKI, CKD, and transplantation. Accumulating evidence points toward lymphangiogenesis being possibly harmful in AKI and kidney allograft rejection, which would potentially position lymphatics as another target for novel therapies to improve outcomes. However, the extent to which lymphangiogenesis is protective rather than maladaptive in the kidney in various settings remains poorly understood and thus an area of active research.

Abstract 2103: Immunophenotyping of extracellular vesicles isolated from lymphatic exudate identifies CD24 as a marker for lymph node metastases in oral cavity squamous cell carcinoma

Abstract Background: Squamous cell carcinoma of the oral cavity (OSCC) is debilitating and fatal with a high risk of lymph node metastases. One of the key problems is that currently there is no robust molecular biomarker to determine whether the tumor has spread or is at risk of spreading to the regional lymph nodes, the first echelon that OSCC spreads to. Extracellular vesicles (EVs) are functional nanoparticles that are bound by natural protein-lipid bilayers. They are deliberately packaged and proactively secreted by all types of cells and serve as intercellular communication vehicles. Tumors release an excess of EVs which are loaded with the molecular and genetic cargo of the parental tumor cells. Methods: Following ethical approval and written informed consent, pre-surgical plasma as well as matched post-surgical lymphatic drainage fluid were collected from 20 OSCC patients (nodal disease n=6, no node n=14), and control plasma was also collected from healthy donors. EVs were isolated by ultracentrifugation, quantified and characterized by nanoparticle tracking analysis, transmission electron microscopy, and western blot. Immune profiling of 37 EV surface markers was determined using flow cytometry with the MACSPlex Exosome kit. Results: Lymphatic exudate was greatly enriched in EVs in comparison to plasma. Common EV tetraspanins, platelet markers, cancer-related proteins, as well as characteristic markers of immune cells were detected in both plasma and lymphatic exudate EVs based on normalized median fluorescence intensity (nMFI). The presence of OSCC led to an altered composition of blood cell-derived EVs. Elevated levels of CD3, CD25, and CD86, and cancer stem cell marker CD133 were observed in the plasma EVs. However, there were no significant changes in EV markers in plasma between patients with and without lymph node metastasis. Notably, EVs isolated from plasma and lymphatic exudate revealed completely divergent signatures of EV surface markers. Among them, cancer-related protein CD24 was found to be the most highly expressed EV marker in lymphatic exudate from patients with nodal disease (nMFI 1.28±0.50 versus 0.84±0.32, p<0.05) and likewise correlated with advanced disease stages (nMFI 1.17±0.40 versus 0.70±0.27, p<0.05). Conclusions: Plasma-derived EVs are promising non-invasive biomarkers for cancer detection. This study shows that lymphatic exudate is a better source of EV biomarkers due to its high abundance of tumor-related factors directly from draining lymph nodes. In addition, CD24, as reported, is indicative of an immunosuppressive tumor microenvironment and it may serve as a predictor for lymph node metastases and progression of OSCC. These findings shed new light on the understanding of OSCC metastases and provide impetus for consideration of CD24-based liquid biopsy testing and possibly novel immunotherapy. Citation Format: Xinyu Qu, Leanne Lee Leung, Cherrie Wing Kei Ng, Hiu Ching Chan, Zigui Chen, Katie Meehan, Xianhai Zeng, Jason Ying Kuen Chan. Immunophenotyping of extracellular vesicles isolated from lymphatic exudate identifies CD24 as a marker for lymph node metastases in oral cavity squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2103.

Rosuvastatin acts on the lymphatic system to improve atherosclerosis

Objective: To investigate whether rosuvastatin acts on lymphatic system and influences lymphatic system-mediated reverse cholesterol transport to play an anti-atherosclerosis role. Methods: Forty-eight apolipoprotein E-/- mice fed a high fat diet were used to construct the atherosclerosis model. They were randomly divided into 4 groups with 12 rats in each group. They were treated with rosuvastatin, vascular endothelial growth factor-C (VEGF-C) and rosuvastatin+VEGF-C inhibitors as experimental group, and no intervention measures were given in control group. After 8 weeks, aortic plaque area, high density lipoprotein cholesterol (HDL-C) content in lymph fluid, the function of popliteal lymphatic drainage of peripheral Evans blue, and the ability of lymphatic system to transport peripheral cell membrane red fluorescent probes to label high-density lipoprotein (HDL) were detected. Subsequently, the effects of rosuvastatin on proliferation, migration and tubular function of lymphoendothelial cells and the expression of scavenger receptor class B type 1 (SR-B1) on lymphoendothelial cells at different concentrations were detected. Results: Compared with the control group, Rosuvastatin and VEGF-C could reduce the area of aortic atherosclerotic plaque (P<0.05). In addition to rosuvastatin plus VEGF-C inhibitor, the intra-aortic plaque area increased (P<0.05). Compared with the control group, Rosuvastatin could increase the content of HDL-C in lymphatic fluid (P<0.05), enhance the drainage function of lymphatic vessels, and enhance the capacity of HDL in the transport tissue fluid of lymphatic system. Compared with the control group, VEGF-C increased the content of HDL-C in mouse lymph fluid (P<0.01), enhanced the drainage function of popliteal lymphatic canal, and enhanced the ability of lymphatic system to transport HDL. With the addition of VEGF-C inhibitor on the basis of rosuvastatin, the content of HDL-C in lymph fluid was reduced, the drainage of popliteal lymphatic canal was interrupted, and the ability of lymphatic system to transport HDL was reduced. Western blotting showed that rosuvastatin increased the protein expression of SR-B1. Conclusion: Rosuvastatin can promote the proliferation, migration and tube formation of lymphatic endothelial cells. At the same time, SR-B1 expression on lymphatic endothelial cells is promoted, thus enhancing the lymphatic system mediated cholesterol reversal transport and playing the role of anti-atherosclerosis.

Imaging and Interventions for Lymphatic and Lymphatic-related Disorders.

The lymphatic system is critical in fluid balance homeostasis. Yet, until recently, lymphatic imaging has been outside of mainstream medicine due to a lack of robust imaging and interventional options. However, during the last 20 years, both clinical lymphatic imaging and interventions have shown dramatic advancement. The key to imaging advancement has been the interstitial delivery of contrast agents through lymphatic-rich tissues. These techniques include intranodal lymphangiography and dynamic contrast-enhanced MR lymphangiography. These methods provide the ability to image and recognize lymphatic anatomy and pathologic conditions. Percutaneous thoracic duct catheterization and embolization became the first widely accepted interventional technique for the management of chyle leaks. Advances in interstitial lymphatic embolization, as well as liver and mesenteric lymphatic interventions, have broadened the scope of possible lymphatic interventions. Also, recent techniques of lymphatic decompression allow for the treatment of a variety of lymphatic disorders. Finally, immunologic studies of central lymphatic fluid reveal the potential of lymphatic interventions on immunity. These advances herald an exciting new chapter for lymphatic imaging and interventions in the coming years.

Validation and efficacy of 'pure' venous lymph node flap in a rat lymphoedema model.

The venous lymph node flap (vLNF) is regarded as an arterialized or 'pure' venous flap, subject to which type of vessel provides blood supply to the flap. However, whether the vLNF drainage function is maintainable or sufficient for the treatment of lymphoedema is yet to be elucidated. In this study, the venous flow-through lymph node flaps or 'pure' vLNF was investigated in a rat's tail (lymphoedema model), and the therapeutic effect was analysed to clarify its potential value for the treatment of lymphoedema. The subjects (rats) were divided into three groups, the experimental group ('pure' vLNF group), the control group (lymphoedema group), and the baseline group. The tail circumference and diameter were recorded for 5 weeks after the operation. Five weeks after surgery, the lymphatic drainage function was evaluated using indocyanine green (ICG) lymphography, while the skin thickness and collagen layer were assessed by histological analysis. The average diameter and circumference of the experimental group were significantly shorter than the control group (p < 0.01). When comparing ICG lymphography within the three groups, the baseline group showed that the ICG entered the inguinal lymph node and appeared in the liver while the ICG still accumulated in the oedematous tail with no fluorescence emerging in other parts of the rat's body, in the control group. In the experimental group, the ICG entered the vLNF via the flap junction with the rat's tail and fluorescence emerging in the rat's liver. When comparing the three groups above with the control group, the experimental group's skin and collagen layer thickness were significantly improved (p < 0.01). Lymphatic recanalization was shown to appear between 'pure' vLNF and the recipient area. Moreover, vLNF was shown to drain lymph fluid via the venous system, thus relieved swelling and fibrosis of the lymphoedematous tissue. Consequently, this could represent a new potential surgical approach for the treatment of lymphoedema.

Successful percutaneous transhepatic lymphangiography and embolization for intractable hepatic lymphorrhea after laparoscopic distal gastrectomy: a case report

BackgroundHepatic lymphorrhea is a rare and serious complication of surgery for digestive tract cancers and is thought to occur as a result of lymph node dissection of the hepatoduodenal ligament. This complication results in the accumulation of lymphatic fluid, which may in turn lead to nutritional disorders, immune deficiency, and circulation insufficiency. However, there is currently no standard strategy for treating this condition.Case presentationA 49-year-old woman with alcoholic liver damage underwent laparoscopic distal gastrectomy with lymph node dissection for early gastric cancer. Abundant ascites persisted postoperatively, and the fluid was suspected to indicate hepatic lymphorrhea. The patient was re-admitted on postoperative day 26 due to the onset of a brain infarction caused by dehydration. Various conservative treatments for hepatic lymphorrhea were ineffective. She underwent percutaneous transhepatic lymphangiography and embolization on postoperative day 81, with obvious effect. Computed tomography images demonstrated complete disappearance of ascites.ConclusionsPostoperative hepatic lymphorrhea is a rare and serious complication of radical surgery for digestive tract cancers. The current case suggests that percutaneous transhepatic lymphangiography and embolization may be a rational treatment option when conservative treatments fail.

Current diagnostic and therapeutic strategies for the management of lymphatic insufficiency in patients with hypoplastic left heart syndrome.

Children with hypoplastic left heart syndrome share unique hemodynamic features that alter lymphatic integrity at all stages of palliation. Lymphatic congestion is almost universal in this patient group to some extent. It may lead to reversal of lymphatic flow, the development of abnormal lymphatic channels and ultimately decompression and loss of protein rich lymphatic fluid into extra lymphatic compartments in prone individuals. Some of the most devastating complications that are associated with single ventricle physiology, notably plastic bronchitis and protein losing enteropathy, have now been proven to be lymphatic in origin. Based on the new pathophysiologic concept new diagnostic and therapeutic strategies have recently been developed. Dynamic contrast magnetic resonance lymphangiography is now mainstay in diagnosis of lymphatic insufficiency and allows a thorough assessment of anatomy and function of the main lymphatic compartments through intranodal, intrahepatic and intramesenteric lymphatic imaging. Contrast enhanced ultrasound can evaluate thoracic duct patency and conventional fluoroscopic lymphangiography has been refined for evaluation of patients where magnetic resonance imaging cannot be performed. Novel lymphatic interventional techniques, such as thoracic duct embolization, selective lymphatic duct embolization and liver lymphatic embolization allow to seal abnormal lymphatic networks minimally invasive and have shown to resolve symptoms. Innominate vein turn-down procedures, whether surgical or interventional, have been designed to reduce lymphatic afterload and increase systemic preload effectively in the failing Fontan circulation. Outflow obstruction can now be managed with new microsurgical techniques that create lympho-venous anastomosis. Short term results for all of these new approaches are overall promising but evidence is sparse and long-term outcome still has to be defined. This review article aims to summarize current concepts of lymphatic flow disorders in single ventricle patients, discuss new emerging diagnostic and therapeutic strategies and point out lacks in evidence and needs for further research on this rapidly growing topic.

A Novel Dressing Composed of Adipose Stem Cells and Decellularized Wharton’s Jelly Facilitated Wound Healing and Relieved Lymphedema by Enhancing Angiogenesis and Lymphangiogenesis in a Rat Model

Lymphedema causes tissue swelling due to the accumulation of lymphatic fluid in the tissue, which delays the process of wound-healing. Developing effective treatment options of lymphedema is still an urgent issue. In this study, we aim to fabricate tissue-engineered moist wound dressings with adipose stem cells (ASCs) and decellularized Wharton's jelly (dWJ) from the human umbilical cord in order to ameliorate lymphedema. Rat ASCs were proliferated and an apparent layer was observed on dWJ at day 7 and 14. A rat tail lymphedema model was developed to evaluate the efficacy of the treatment. Approximately 1 cm of skin near the base of the rat tail was circularly excised. The wounds were treated by secondary healing (control) (n = 5), decellularized Wharton's jelly (n = 5) and ASC-seeded dWJ (n = 5). The wound-healing rate and the tail volume were recorded once a week from week one to week five. Angiogenesis and lymphangiogenesis were assessed by immunochemistry staining with anti-CD31 and anti-LYVE1. The results showed that the wound-healing rate was faster and the tail volume was lesser in the ASC-seeded dWJ group than in the control group. More CD31+ and LYVE-1+ cells were observed at the wound-healing area in the ASC-seeded dWJ group than in the control group. This proves that tissue-engineered moist wound dressings can accelerate wound-healing and reduce lymphedema by promoting angiogenesis and lymphangiogenesis.

Cytological and Biochemical Analyses of Lymphatic Fluid from Patients with Lymphatic Malformations.

Background: Intracystic hemorrhage from lymphangiomas is a common phenomenon in lymphatic malformations (LMs); however, little is known about the associated compositional changes in the lymphatic fluid. Materials and Methods: We prospectively collected lymphatic fluid from children with LMs. Lymphatic fluid was divided depending on the bleeding status into the bleeding and nonbleeding groups. The fluid was subjected to cytological and biochemical analyses to determine protein and cytokine levels. The Mann-Whitney U test was used to compare the two groups. Results: There were significant differences in the levels of interleukin (IL)-6, IL-10, and glucose, and the percentage of white blood cells between the bleeding and nonbleeding groups. There was no significant difference in chlorine and protein content; white blood cell count; and IL-2, IL-4, tumor necrosis factor α, and interferon γ levels between the two groups. Conclusion: Lymphatic fluid is less stable in bleeding LMs than in non-bleeding LMs and is prone to inflammatory reactions. The inflammatory reaction in lymphatic fluid does not stimulate the cytokine storm in blood. The inflammatory reaction due to LMs does not affect the contents of protein and chlorine in lymphatic cyst fluid.

Lymphatic flow velocity is a predictor of functional lymphatic vessels for lymphaticovenous anastomosis

Indocyanine green (ICG) lymphography is widely used to localize functional lymphatic vessels for lymphaticovenous anastomosis (LVA); however, flow velocity is rarely assessed. We aimed to evaluate the correlation between lymphatic flow velocity and the presence of functional lymphatic vessels. Data of a total of 924 lymphatic vessels from 273 lymphedema patients who underwent LVA between July 2018 and December 2020 were retrospectively reviewed. Lymph flow velocity was defined by considering the most proximal anatomic location enhanced by ICG at 30 min after injection and categorized into four groups; grade 1 (foot or hand), grade 2 (below knee or elbow), grade 3 (at/above knee or eblow), or grade 4 (axilla or groin). The presence of functional lymphatic vessels, which showed lymphatic fluid flow when the vessels were cut for anastomosis, was compared between the four groups. A higher rate of functional lymphatic vessels was observed among lymphatic vessels with grade 3 or 4 flow velocity compared with those with grade 1 or 2 flow velocity (67.5% vs. 44.5%; p < 0.001). These findings were consistent with the observations for lymphatic vessels with a non-linear pattern in ICG lymphography (59.4% vs. 26.5%; p < 0.001). The rate of completion of LVA at surgical sites in extremities with grade 3 or 4 flow velocity was 88.1% compared with 65.8% in extremities with grade 1 or 2 velocity (p < 0.001). Lymph flow velocity grading can be a simple and easy adjunctive method to determine indication for LVA in extremities with lymphedema.

Thoracic duct leakage in a patient with Type B-Non-Hodgkin lymphoma treated with transvenous retrograde access embolization: a case report.

Thoracic duct (TD) is the largest lymphatic vessel in the body and drains the lymph at the junction between the left subclavian and jugular veins. Chylothorax (CTX) represents an accumulation of lymphatic fluid in the pleural space. We present a case of a 65 years-old man with an histologically diagnosed mediastinal type B non-Hodgkin Lymphoma, treated with chemo-immunotherapy. CT scan during follow up showed significant left side pleural effusion, amounting to 2.8 litres after drainage. Conservative treatment with low fat parenteral nutrition was started without reduction of drainage output, then lymphangiography (LP) with Lipiodol was performed demonstrating a leak in the distal TD. CTX increased in the following days, and a further LP was performed. Using transvenous retrograde access we catheterized TD at the left subclavian jugular veins using a microcatheter. The leak was treated with multiple conventional and controlled delivery microcroils and cyanoacrylate, obtaining complete embolization without residual leak.

Development and application of a novel cervical lymph collection method to assess lymphatic transport in rats.

Background: Fluids, solutes and immune cells have been demonstrated to drain from the brain and surrounding structures to the cervical lymph vessels and nodes in the neck via meningeal lymphatics, nasal lymphatics and/or lymphatic vessels associated with cranial nerves. A method to cannulate the efferent cervical lymph duct for continuous cervical lymph fluid collection in rodents has not been described previously and would assist in evaluating the transport of molecules and immune cells from the head and brain via the lymphatics, as well as changes in lymphatic transport and lymph composition with different physiological challenges or diseases. Aim: To develop a novel method to cannulate and continuously collect lymph fluid from the cervical lymph duct in rats and to analyze the protein, lipid and immune cell composition of the collected cervical lymph fluid. Methods: Male Sprague-Dawley rats were cannulated at the carotid artery with or without cannulation or ligation at the cervical lymph duct. Samples of blood, whole lymph and isolated lipoprotein fractions of lymph were collected and analyzed for lipid and protein composition using commercial kits. Whole lymph samples were centrifuged and isolated pellets were stained and processed for flow cytometry analysis of CD3+, CD4+, CD8a+, CD45R+ (B220) and viable cell populations. Results: Flow rate, phospholipid, triglyceride, cholesterol ester, free cholesterol and protein concentrations in cervical lymph were 0.094 ± 0.014mL/h, 0.34 ± 0.10, 0.30 ± 0.04, 0.07 ± 0.02, 0.02 ± 0.01 and 16.78 ± 2.06mg/mL, respectively. Protein was mostly contained within the non-lipoprotein fraction but all lipoprotein types were also present. Flow cytometry analysis of cervical lymph showed that 67.1 ± 7.4% of cells were CD3+/CD4+ T lymphocytes, 5.8 ± 1.6% of cells were CD3+/CD8+ T lymphocytes, and 10.8 ± 4.6% of cells were CD3-/CD45R+ B lymphocytes. The remaining 16.3 ± 4.6% cells were CD3-/CD45- and identified as non-lymphocytes. Conclusion: Our novel cervical lymph cannulation method enables quantitative analysis of the lymphatic transport of immune cells and molecules in the cervical lymph of rats for the first time. This valuable tool will enable more detailed quantitative analysis of changes to cervical lymph composition and transport in health and disease, and could be a valuable resource for discovery of biomarkers or therapeutic targets in future studies.

Abstract PR017: Differences in melanoma lipid metabolism among distinct metastatic sites

Abstract Metastasis is a very inefficient process in which few disseminated cancer cells survive. We developed a patient-derived xenograft assay in which melanomas engraft and spontaneously metastasize (Quintana Nature 456:593-598). Using this assay, we discovered that melanoma cells experience a spike in reactive oxygen species during metastasis and that distant metastasis is limited by oxidative stress (Piskounova Nature 527:186-191). Successfully metastasizing cells undergo reversible metabolic changes during metastasis that increase oxidative stress resistance. For example, melanoma cells in lymph experience less oxidative stress and form more metastases than melanoma cells in the blood (Ubellacker Nature 585:113-120). This is true in both patient-derived melanomas growing in immunocompromised mice and mouse melanomas growing in syngeneic immunocompetent mice. Cells metastasizing through blood, but not lymph, appear to undergo ferroptosis, a form of cell death marked by lipid oxidation. Multiple differences between lymph fluid and blood plasma may contribute to this difference in oxidative stress, including higher levels of oleic acid in lymph. Oleic acid is a monounsaturated fatty acid that protects melanoma cells from ferroptosis, apparently by reducing the abundance of oxidizable polyunsaturated fatty acids (PUFAs) in phospholipids. In the current study we found that some PUFAs sensitize melanoma cells to lipid peroxidation and cell death more than others. Moreover, we analyzed the lipid profiles of common sites of distant metastasis – liver, brain, kidney, pancreas, bone marrow, lung, and liver – and found that the PUFA compositions of these tissues varies significantly. This raises the possibility that metastasizing melanoma cells may incorporate more PUFAs into their phospholipid membranes and exhibit increased ferroptosis sensitivity in some metastatic sites as compared to others. We found that melanoma cells isolated from metastatic sites by flow cytometry had higher levels of PUFAs in their phospholipid membranes compared to those isolated from subcutaneous tissues. These cells isolated from distal sites also exhibited higher levels of lipid ROS and contained higher levels of oxidized lipid species compared to subcutaneous tumors. These data suggest that the lipid metabolism of metastatic tumors reflects both lipid availability in the metastatic site as well as cell-intrinsic differences in lipid metabolism, both of which contribute to lipid ROS. Both cell-intrinsic and cell-extrinsic lipid metabolism likely influence the ability of melanoma cells to survive in distinct metastatic sites during metastasis. Citation Format: Thomas P. Mathews, Sarah Muh, Arin B. Aurora, Sean J. Morrison. Differences in melanoma lipid metabolism among distinct metastatic sites [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr PR017.

Oral Sildenafil in the Treatment of Cystic Hygroma in Infant

Introduction: Cystic hygroma is a type of cyst that occurs when lymphatic fluid collects in the lymph nodes which occurs in infants and can cause significant swelling in the neck or other parts of the body. Sildenafil is a possible medication that can be used to treat erectile dysfunction in infant. Aim of the study: The aim of this study was to assess the effectiveness of oral sildenafil in the treatment of cystic hygroma in infant. Methods: This retrospective study was conducted from January 2020 to June 2022 in The Division of Pediatric Surgery, Bangladesh Shishu Hospital &amp; Institute and three other private clinics at Dhaka. Total 55 patients (aged between 2 months to 1 year) with cystic hygroma were included in the study. Result: In the present study, the median (Range) age of 55 patients was 6.2 (2-12) months. The median (Range) weight of 55 patients was 6.5 (4-12) kg. In this study, majority (60%) of the patients were male. More than half (52.73%) patients had the lesion on neck. The median (Range) pretreatment size of lesion was 13.30 (9.88-16.93) cm2. The median pretreatment size of lesion was 13.30 cm2 and median post treatment size of lesion was 0.00 cm2. There was highly significant difference (p &lt;0.001) between pre-treatment and post-treatment size. In this study, excellent response was found in 69.09% patients, good response was found in 21.82% patients. In the current study, 3 patients (5.45%) had cough, 4 patients (7.27%) had fever, 2 patients (3.64%) had rhinorrhoea and 3 patients (5.45%) had vomiting. In the current study, 12 (21.82%) patients had adverse effects. Cost of sildenafil per tablet was 34 cents (100 cents =1 $). Conclusion: Oral sildenafil resulted excellent size regression and low adverse effects for the treatment of cystic hygroma. It was also very cost effective and convenient to administer.

C-JUN-mediated transcriptional responses in lymphatic endothelial cells are required for lung fluid clearance at birth

Fluid clearance mediated by lymphatic vessels is known to be essential for lung inflation and gas-exchange function during the transition from prenatal to postnatal life, yet the molecular mechanisms that regulate lymphatic function remain unclear. Here, we profiled the molecular features of lymphatic endothelial cells (LECs) in embryonic and postnatal day (P) 0 lungs by single-cell RNA-sequencing analysis. We identified that the expression of c-JUN is transiently upregulated in P0 LECs. Conditional knockout of Jun in LECs impairs the opening of lung lymphatic vessels at birth, leading to fluid retention in the lungs and neonatal death. We further demonstrated that increased mechanical pressure induces the expression of c-JUN in LECs. c-JUN regulates the opening of lymphatic vessels by modulating the remodeling of the actin cytoskeleton in LECs. Our study established the essential regulatory function of c-JUN-mediated transcriptional responses in facilitating lung lymphatic fluid clearance at birth.

SiRNA delivery to lymphatic endothelial cells via ApoE-mediated uptake by lipid nanoparticles.

Systemically administered lipid nanoparticles (LNPs) are complexed with Apolipoprotein E (ApoE) in the bloodstream, and the complex is subsequently largely taken up by hepatocytes. Based on a previous report showing that, like blood, lymph fluid also contains ApoE, and that LECs, in turn, expresses a low density-lipoprotein receptor (LDLR), which is the receptor responsible for the ApoE-bound LNP, we hypothesized that subcutaneously administered LNPs would be taken up by LECs via an ApoE-LDLR pathway. Our in vitro studies using immortal LECs that we established in a previous study showed that LEC indeed took up LNPs in an ApoE-dependent manner. We then reported on the development of LNPs that target the lymphatic endothelium for in vivo siRNA delivery after subcutaneous administration. The key to success for in vivo LEC targeting is that the surface needs to be modified with a high density of polyethylene glycol (PEG)-conjugated lipids with short acyl chains (C14). The LNPs were drained into the lymphatic system, and then accumulated in lymphatic endothelial cells in an ApoE-dependent manner, most likely after the release of the PEG-lipid. Subcutaneous administration of optimized LNPs containing encapsulated siRNA against VEGFR3, a marker of LECs, significantly inhibited the expression of VEGFR3. These findings are the first report of a simple straightforward strategy for targeting lymphatic endothelial cells by using ionizable lipid-formulated LNPs.

An Unusual Case Report of Adult Cervical Cystic Hygroma

Cystic Lymphangioma (CL) is a rare condition that is typically observed as a congenital malformation in the head and neck. These congenital lesions usually appear before the age of two. Adult occurrences are extremely unusual, and only a few cases have been mentioned in the literature. CL is characterised by the collection of an epithelial lining of lymphatic fluid, which can be caused by lymphatic abnormalities, trauma, or surgery. In the present case report, the authors present the findings of a 49-year-old adult female who had painless, gradually increasing left-sided neck swelling for three years, with no other relevant medical history. The initial ultrasound revealed a large cystic lesion extending from the left sternocleidomastoid muscle to the left submandibular gland. The lesion measured 8×7×3 cm and exhibited narrow septae and loculation at the periphery. Avascularity was noted within the cyst wall and interlobular septae. The diagnosis favoured macrocytic lymphangioma/Congenital Lymphangioma (CH). During the surgical procedure, the cyst was dissected, and it was found to be located over the internal jugular vein, carotid artery, and spinal accessory nerve. The cyst was completely removed, and a surgical drain was left in place. The wound was sutured in two layers, and a sterile dressing was applied. The drain was removed on the third postoperative day, and the dressing with sutures was removed on the seventh postoperative day, revealing a healthy wound. The diagnosis was confirmed by histopathological examination {Haematoxylin and Eosin (H&amp;E)} of the tissue specimen, which showed flattened epithelium lining the inside of the cyst. The cyst contained dilated gaps in a fibrocollagenous stroma infiltrated by lymphocytes. Therefore, CH can be considered as a differential diagnosis for head and neck masses in adults.

Antibodies and immune-enhancing compounds: A comprehensive review

The immune system is a complex network of cells, tissues, and organs that safeguards the body against harmful environmental factors and maintains overall health. Its primary function is to mount an immune response to prevent infections and ensure the body's well-being. This defense mechanism involves the collaboration of white blood cells, the thymus gland, spleen, lymphatic glands, stem cells, proteins, and antibodies, among other components. White blood cells act as the front line of defense by identifying and eliminating pathogens. The thymus gland plays a crucial role in the development and maturation of T-cells, which are vital for adaptive immunity. The spleen filters the blood and removes old or damaged blood cells, while lymphatic glands serve as reservoirs for immune cells and filter lymph fluid. Stem cells contribute to the continuous renewal of immune cells. Proteins, including antibodies, play a critical role in identifying and neutralizing specific pathogens. Understanding the intricate workings of the immune system, along with the development of immune-enhancing compounds and antibodies, is essential for enhancing our ability to combat infections and maintain overall health.

Vascular Invasion Predicts Recurrence in Stage IA2-IB Lung Adenocarcinoma but not Squamous Cell Carcinoma

BackgroundLymphovascular invasion (LVI) is an adverse prognostic feature in resected stage I non-small cell lung cancer (NSCLC); however, it is unclear if the prognostic significance applies to both lung adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC). Materials and MethodsA retrospective review of H&E-stained slides from surgically resected AJCC 8th ed. stage IA2-IB LUAD (n = 344) and LUSC (n = 102) from two institutions was performed. LVI was defined as either lymphatic (LI) or vascular (VI) invasion. Outcomes were assessed by 5-year recurrence-free survival (RFS) estimates using the Kaplan-Meier method. ResultsThe cohorts of LUAD and LUSC showed no significant differences in 5-year RFS (81% each), stage, age, race, or surgical procedure. The presence of LVI, VI, and LI was predictive of 5-year RFS for LUAD (LVI + 71% vs. LVI - 92%, P < 0.001; VI + 64% vs. VI - 90%, P < 0.001; LI + 75% vs. LI - 84%, P = 0.030) but not LUSC (LVI + 84% vs. LVI - 79%, P = 0.740; VI + 83% vs. VI- 80%, P = 0.852; LI + 84% vs. LI - 81%, P = 0.757). Among LUAD with LVI, VI was a stronger predictor of 5-year RFS than the remaining subset of VI-LI + tumors (64% vs. 87%, P = 004). Subset analysis of LI among LUAD stratified by VI showed no significant prognostic advantage to adding LI for risk stratification (VI-LI + 87% vs. VI-LI - 92%, P = 0.347 & VI+LI + 62% vs. VI + LI- 66%, P = 0.422). VI was present in 36% of LUAD. ConclusionVascular invasion is a strong predictor of recurrence in stage IA2-IB LUAD but not in LUSC. Adjuvant therapy trials should be directed at this subgroup.