Sacral Nodes Research Articles

Whole CNS 3D Cryo-Fluorescence Tomography Shows CSF Clearance along Nasal Lymphatics, Spinal Nerves, and Lumbar/Sacral Lymph Nodes

Unwanted proteins and metabolic waste in cerebral spinal fluid are cleared from the brain by meningeal and nasal lymphatics and the perineural sheath of cranial nerves; however, the distribution and clearance of cerebral spinal fluid (CSF) along the subarachnoid space of the entire spinal cord is not fully understood. Cryo-fluorescence tomography (CFT) was used to follow the movement of tracers from the ventricular system of the brain down through the meningeal lining of the spinal cord and out to the spinal lymphatic nodes. Isoflurane-anesthetized mice were infused into the lateral cerebroventricle with 5.0 µL of quantum dots [QdotR 605 ITKTM amino (PEG)] over two mins. Mice were allowed to recover (ca 2-3 min) and remained awake and ambulatory for 5, 15, 30, 60, and 120 min after which they were euthanized, and the entire intact body was frozen at -80°. The entire mouse was sectioned, and white light and fluorescent images were captured after each slice to produce high resolution three-dimensional volumes. Tracer appeared throughout the ventricular system and central canal of the spinal cord and the entire subarachnoid space of the CNS. A signal could be visualized in the nasal cavity, deep cervical lymph nodes, thoracic lymph nodes, and more superficial submandibular lymph nodes as early as 15 min post infusion. A fluorescent signal could be visualized along the dorsal root ganglia and down the proximal extension of the spinal nerves of the thoracic and lumbar segments at 30 min. There was a significant accumulation of tracer in the lumbar and sacral lymph nodes between 15-60 min. The dense fluorescent signal in the thoracic vertebrae noted at 5- and 15-min post infusion was significantly reduced by 30 min. Indeed, all signals in the spinal cord were ostensibly absent by 120 min, except for trace amounts in the coccyx. The brain still had some residual signal at 120 min. These data show that Qdots with a hydrodynamic diameter of 16-20 nm rapidly clear from the brain of awake mice. These data also clearly demonstrate the rapid distribution and efflux of traces along a major length of the vertebral column and the potential contribution of the spinal cord in the clearance of brain waste.

Feasibility study of direct CT lymphangiography in mice: comparison with interstitial CT/MR lymphangiography

ObjectivesTo establish a CT lymphangiography method in mice via direct lymph node puncture.MethodsWe injected healthy mice (n = 8) with 50 µl of water-soluble iodine contrast agent (iomeprol; iodine concentration, 350 mg/mL) subcutaneously into the left-rear foot pad (interstitial injection) and 20 µl of the same contrast agent directly into the popliteal lymph node (direct puncture) 2 days later. Additionally, we performed interstitial MR lymphangiography on eight mice as a control group. We calculated the contrast ratio for each lymph node and visually assessed the depiction of lymph nodes and lymphatic vessels on a three-point scale.ResultsThe contrast ratios of 2-min post-injection images of sacral and lumbar–aortic lymph nodes were 20.7 ± 16.6 (average ± standard deviation) and 17.1 ± 12.0 in the direct puncture group, which were significantly higher than those detected in the CT or MR interstitial lymphangiography groups (average, 1.8–3.6; p = 0.008–0.019). The visual assessment scores for sacral lymph nodes, lumbar–aortic lymph nodes, and cisterna chyli were significantly better in the direct puncture group than in the CT interstitial injection group (p = 0.036, 0.009 and 0.001, respectively). The lymphatic vessels between these structures were significantly better scored in direct puncture group than in the CT or MR interstitial lymphangiography groups at 2 min after injection (all p ≤ 0.05).ConclusionsIn CT lymphangiography in mice, the direct lymph node puncture provides a better delineation of the lymphatic pathways than the CT/MR interstitial injection method.Key Points•The contrast ratios of 2-min post-injection images in the direct CT lymphangiography group were significantly higher than those of CT/MR interstitial lymphangiography groups.•The visibility of lymphatic vessels in subjective analysis in the direct CT lymphangiography group was significantly better in the direct puncture group than in the CT/MR interstitial lymphangiography groups.•CT lymphangiography with direct lymph node puncture can provide excellent lymphatic delineation with contrast being maximum at 2 min after injection.

Anatomic, computed tomographic, and ultrasonographic assessment of the lymph nodes in presumed healthy adult cats: the abdomen, pelvis, and hindlimb

BackgroundThe computed tomography (CT) and ultrasonography (US) features of lymph nodes of the abdomen, pelvis, and hindlimb in healthy cats are poorly described in the current literature. A prospective anatomic and reference interval study was therefore performed. The lymph nodes of six feline cadavers were identified, and dimensions were measured (length, width, and height). The lymph nodes from 30 healthy adult cats were identified and measured using CT (pre- and postcontrast) and US. The identification and dimensions of the separate lymph nodes were compared between imaging techniques and the anatomic study.ResultsThe identification of lymph nodes was most frequent in CT, and the dimensions were overall larger than those identified and measured in US and the anatomic study. The caudal epigastric and sacral lymph nodes were not identified in the anatomic study. The ischiatic, lumbar aortic, internal iliac, and caudal epigastric lymph nodes were not visualized in US. The height presented the main statistical differences among techniques. The lymph nodes were mainly homogeneous in pre- and postcontrast CT and US images. Some lymph nodes showed a hyperattenuating periphery with a hypoattenuating center (on pre- and postcontrast images) and a hypo-/isoechoic periphery with a hyperechoic center, representing the hilar fat. The lymph nodes were commonly elongated and rounded except for the jejunal lymph nodes, which had an irregular shape.ConclusionsThe assessment of most of the abdominal, pelvic, and hindlimb lymph nodes in the cat is feasible using CT and US, with CT performing best. Factors like the amount of adipose tissue and contrast administration subjectively improved the lymph node visualization and assessment. The measurements and features reported are proposed as reference values.

Effects of increased intracranial pressure on cerebrospinal fluid influx, cerebral vascular hemodynamic indexes, and cerebrospinal fluid lymphatic efflux

The glymphatic-lymphatic fluid transport system (GLFTS) consists of glymphatic pathway and cerebrospinal fluid (CSF) lymphatic outflow routes, allowing biological liquids from the brain parenchyma to access the CSF along with perivascular space and to be cleaned out of the skull through lymphatic vessels. It is known that increased local pressure due to physical compression of tissue improves lymphatic transport in peripheral organs, but little is known about the exact relationship between increased intracranial pressure (IICP) and GLFTS. In this study, we verify our hypothesis that IICP significantly impacts GLFTS, and this effect depends on severity of the IICP. Using a previously developed inflating balloon model to induce IICP and inject fluorescent tracers into the cisterna magna, we found significant impairment of the glymphatic circulation after IICP. We further found that cerebrovascular occlusion occurred, and cerebrovascular pulsation decreased after IICP. IICP also interrupted the drainage of deep cervical lymph nodes and dorsal meningeal lymphatic function, enhancing spinal lymphatic outflow to the sacral lymph nodes. Notably, these effects were associated with the severity of IICP. Thus, our findings proved that the intensity of IICP significantly impacts GLFTS. This may have translational applications for preventing and treating related neurological disorders.

Water-Soluble High-Quality Ag2Te Quantum Dots Prepared by Mutual Adaptation of Synthesis and Surface Modification for In Vivo Imaging.

Near-infrared (NIR) in vivo fluorescence imaging has exhibited the distinct advantage of high optical resolution at deeper penetration into biological tissues. Ag2Te quantum dots (QDs), with a relatively narrow band gap, show great promise for fluorescence emission at long wavelengths in the second near-infrared (NIR-II) window for bioimaging. However, existing Ag2Te QDs have severely hindered the application of in vivo bioimaging due to their poor fluorescence brightness and stability, so it is important to prepare Ag2Te QDs with high quantum yield and stability as well as high biocompatibility in the NIR-II window. Herein, we designed an integrated method for the preparation of water-soluble Ag2Te QDs by mutual adaptation of QD synthesis and surface modification. We first synthesized high-quality Ag2Te QDs with different NIR-II emission wavelengths and the photoluminescence quantum yields (PLQYs) up to 6.51% by rapidly injecting the TBP-Te precursor into a hot solvent to form a highly fluorescent Ag2Te core. Then water-dispersible Ag2Te QDs were obtained by direct exchange of the hydrophobic Ag2Te QD surface ligands with thiol ligands. The PLQY of the water-soluble Ag2Te QDs obtained by this method can still be maintained at 4.94%. With these highly bright and stable Ag2Te QDs, the abdominal vessels, hindlimb arterial vessels, venous vessels, sacral lymph nodes, and tumor vessels were visualized non-invasively in vivo in the NIR-II window in mice. The results demonstrate that the integrated strategy of QD synthesis and modification provides valuable technical support for further in-depth applications of Ag2Te QDs.

The underestimated posterior lymphatic drainage of the prostate: An historical overview and preliminary anatomical study on cadaver.

The evidence of pelvic lymph node metastases after radical prostatectomy (RP) with pelvic lymph node dissection (PLND) is one of the strongest prognostic factors for poor oncologic outcome. The extent of PLND, although representing a crucial step in RP, is still controversial. Currently, there is a critical drawback in clinical practice due to the lack of congruence between the known lymphatic drainage and cancer dissemination despite defined management by a surgical approach. We hypothesized the existence of alternative pathways for the lymphatic drainage of the prostate currently not considered in clinical daily practice. We carried out a literature review of the anatomic description of nodal drainage of prostate reported by online databases (MEDLINE/PubMed, EBSCO, Web of Science, Ovid, and Scopus) and the original texts since the 18th century, with an additional anatomical dissection on a human cadaver to confirm theoretical data. The anatomical dissection study converged with the historical anatomical treatises in describing three groups of lymphatics devoted to carrying out prostatic nodal drainage. Apart from the ascending ducts from the cranial gland leading to the external iliac nodes; the lateral ducts leading to the hypogastric nodes; small lymphatic vessels from the posterior surface of the prostate, directed to the pararectal lymphatic plexus, in the direction of the lateral sacral lymph nodes and those at the sacral promontory (ie, pararectal and presacral lymph nodes) were observed. Our preliminary findings demonstrate that lymphatic drainage of the prostate extends beyond standard nodal templates actually considered in surgical daily practice, despite the knowledge reported by historical anatomical treatises. Further anatomical and experimental evidence are needed to investigate anatomical variability in humans, as well as to add more topographical details.

Computed tomographic appearance of abdominal lymph nodes in healthy cats.

BackgroundComputed tomography (CT) is commonly used in veterinary medicine and plays an important role in disease identification and cancer staging. Identification of abnormal abdominal lymph nodes is important for clinical, therapeutic, and prognostic decision making. No published study describes the CT appearance of abdominal lymph nodes in healthy cats.Hypothesis/ObjectivesAll abdominal lymph centers will be identified on CT with the majority of lymph nodes being elongated and homogenously contrast enhancing.AnimalsSixteen healthy cats without clinical or biochemical evidence of disease.MethodsPrecontrast and postcontrast CT images of sedated healthy cats were used to identify the presence and descriptive characteristics of intra‐abdominal lymph nodes. These assessments then were compared with patient characteristics to identify possible correlations.ResultsAbdominal lymph nodes were readily identified on CT with caudal mesenteric, colic, hepatic, inguinal, and pancreaticoduodenal lymph nodes identified in 16/16 cats. Lymph node size and shape varied among lymph centers with nearly all lymph nodes homogeneously contrast enhancing in 515/525. Significant negative correlations were identified between age and length (P = .0166) and width (P = .0387) of abdominal lymph nodes as well as age and number of sacral lymph nodes (P = .0493). Intranodal fat was present in 18/525 lymph nodes.Conclusions and Clinical ImportanceCT readily permitted identification and characterization of feline abdominal lymph nodes. This study provides subjective and objective data on the CT characteristics of abdominal lymph nodes in 16 healthy cats, with younger cats having larger abdominal lymph nodes and a higher number of sacral lymph nodes.

Frequency of an accessory popliteal efferent lymphatic pathway in dogs.

Staging and therapeutic planning for dogs with malignant disease in the popliteal lymph node are based on the expected patterns of lymphatic drainage from the lymph node. The medial iliac lymph nodes are known to receive efferent lymph from the popliteal lymph node; however, an accessory popliteal efferent pathway with direct connection to the sacral lymph nodes has also been less frequently reported. The primary objective of this prospective, anatomic study was to describe the frequency of various patterns of lymphatic drainage of the popliteal lymph node. With informed client consent, 50 adult dogs with no known disease of the lymphatic system underwent computed tomographic lymphography after ultrasound-guided, percutaneous injection of 350mg/ml iohexol into a popliteal lymph node. In all 50 dogs, the popliteal lymph node drained directly to the ipsilateral medial iliac lymph node through multiple lymphatic vessels that coursed along the medial thigh. In 26% (13/50) of dogs, efferent vessels also drained from the popliteal lymph node directly to the internal iliac and/or sacral lymph nodes, coursing laterally through the gluteal region and passing over the dorsal aspect of the pelvis. Lymphatic connections between the right and left medial iliac and right and left internal iliac lymph nodes were found. Based on our findings, the internal iliac and sacral lymph nodes should be considered when staging or planning therapy for dogs with malignant disease in the popliteal lymph node.

PEG-poly(L-lysine)-based polymeric micelle MRI contrast agent: Feasibility study of a Gd-micelle contrast agent for MR lymphography.

To investigate the feasibility of polymeric micelle of poly(ethyleneglycol) (PEG)-b-poly(L-lysine-DOTA) (Gd-micelle) as a contrast agent for magnetic resonance lymphography (MRL). Twenty-four female BALB/c mice were randomly divided into four groups of six mice each. Among them, mice of two groups were injected of complete Freund's adjuvant to obtain inflamed lymph nodes. We subcutaneously injected 0.5 μmol Gd per mouse of Gd-micelle or gadofluorine P in the right rear footpad. Identical 3D T1 -weighted gradient-echo imaging (1T MRI system) were subsequently obtained to create time-intensity curves of the right popliteal, sacral, and lumbar-aortic lymph nodes and to measure the contrast ratios (CRs). The peak CR, area under the curve (AUC), and elimination half-life (T1/2 ) of CR of the popliteal lymph node were assessed by two-way factorial analysis of variance. We also performed a qualitative assessment of normal and inflamed lymph node at three timepoints. The mean peak CR of Gd-micelle was 2.64 and 1.89 for gadofluorine P in normal mice, and 3.48 and 2.73 in the inflamed lymph node. Statistically, peak CR was higher for Gd-micelle (P = 0.004). In addition, the AUC was larger (P < 0.001) and T1/2 was longer (P < 0.001) for Gd-micelle. In qualitative assessment, Gd-micelle demonstrated the same or higher scores in every lymph node, and demonstrated a higher score in lumbar-aortic lymph node of a 360-minute image (P = 0.006) and in inflamed lymph node of a 360-minute image (P = 0.009). Compared to gadofluorine P, Gd-micelle showed higher and more prolonged enhancement in MRL imaging in normal and inflamed lymph nodes. 1 Technical Efficacy: Stage 1 J. Magn. Reson. Imaging 2018;47:238-245.

INDIRECT COMPUTED TOMOGRAPHIC LYMPHOGRAPHY FOR ILIOSACRAL LYMPHATIC MAPPING IN A COHORT OF DOGS WITH ANAL SAC GLAND ADENOCARCINOMA: TECHNIQUE DESCRIPTION.

Sentinel lymph node mapping can help to direct surgical oncologic staging and metastatic disease detection in patients with complex lymphatic pathways. We hypothesized that indirect computed tomographic lymphography (ICTL) with a water-soluble iodinated contrast agent would successfully map lymphatic pathways of the iliosacral lymphatic center in dogs with anal sac gland carcinoma, providing a potential preoperative method for iliosacral sentinel lymph node identification in dogs. Thirteen adult dogs diagnosed with anal sac gland carcinoma were enrolled in this prospective, pilot study, and ICTL was performed via peritumoral contrast injection with serial caudal abdominal computed tomography scans for iliosacral sentinel lymph node identification. Technical and descriptive details for ICTL were recorded, including patient positioning, total contrast injection volume, timing of contrast visualization, and sentinel lymph nodes and lymphatic pathways identified. Indirect CT lymphography identified lymphatic pathways and sentinel lymph nodes in 12/13 cases (92%). Identified sentinel lymph nodes were ipsilateral to the anal sac gland carcinoma in 8/12 and contralateral to the anal sac gland carcinoma in 4/12 cases. Sacral, internal iliac, and medial iliac lymph nodes were identified as sentinel lymph nodes, and patterns were widely variable. Patient positioning and timing of imaging may impact successful sentinel lymph node identification. Positioning in supported sternal recumbency is recommended. Results indicate that ICTL may be a feasible technique for sentinel lymph node identification in dogs with anal sac gland carcinoma and offer preliminary data to drive further investigation of iliosacral lymphatic metastatic patterns using ICTL and sentinel lymph node biopsy.

Ultrasound and computed tomography of the iliosacral lymphatic centre in dogs with anal sac gland carcinoma

In this prospective study, we hypothesized that computed tomography (CT) would identify more normal and abnormal iliosacral lymph nodes (LNs) than abdominal ultrasound in dogs with anal sac gland carcinoma (ASGC). Twelve client-owned dogs with ASGC but without distant metastasis were enrolled. Abdominal ultrasound and contrast-enhanced CT scans of the abdomen were obtained. Iliosacral LNs were counted and assessed for location, laterality and size. Significantly (P < 0.00001) more iliosacral LNs were identified with CT (61) than ultrasound (30), including significantly (P = 0.00012) more medial iliac LNs with CT (33) than ultrasound (19). There was no difference in number of internal iliac LNs identified with CT versus ultrasound. Significantly (P = 0.000061) more sacral LNs were identified with CT (15) than ultrasound (0). Ultrasound identified slightly more (7) abnormal iliosacral LNs than CT (5). Contrast CT was able to identify more normal but not more abnormal LNs than ultrasound.

Interstitial MR Lymphography in Mice: Comparative Study with Gadofluorine 8, Gadofluorine M, and Gadofluorine P

We investigated the characteristics and capability of interstitial MR lymphography in mice using gadofluorine 8, gadofluorine M, and gadofluorine P. We injected healthy mice with 0.5 µmol of Gd gadofluorine 8, gadofluorine M, or gadofluorine P subcutaneously into the right rear footpad and assessed the time courses of contrast enhancement in the lymph nodes. Six mice were studied for each contrast agent. We also used gadofluorine M to assess the lymphatic pathway from the right and left rear feet or tail. Contrast enhancement was demonstrated for the right popliteal, sacral, and iliac lymph nodes in all mice 5 minutes after injection of each of the 3 agents and decreased gradually. Enhancement in the lymph nodes was still detectable 30 minutes after injection of gadofluorine 8 or gadofluorine M. Enhancement became obscure sooner after gadofluorine P injection and was mildly stronger with the other 2 contrast agents. Clear differences were found in the hepatobiliary and urinary kinetics of the 3 agents. Gadofluorine M injected into various sites delineated the lymphatic pathway from the site of injection. Interstitial MR lymphography using gadofluorine 8, gadofluorine M, and gadofluorine P offered clear visualization of the lymphatic pathway in healthy mice during a sufficient imaging time window, and allowed repeated assessment of the pathway and clarification of the lymphatic system.

Integrated Lymphography using Fluorescence Imaging and Magnetic Resonance Imaging in Intact Mice

We assessed lymph drainage in living mice by an integrated imaging method using fluorescence imaging (FLI) and magnetic resonance imaging (MRI). Mice were subcutaneously injected with quantum dots and gadofluorine 8 into the right rear footpad. They were fixed on a transparent flat plate and underwent FLI and MRI successively. Small markers were attached to the mouse surface for spatial coregistration, and image fusion of FLIs and MRIs was performed. Two-dimensional fluorescence reflectance imaging was used for FLI. FLI and MRI provided generally consistent results and demonstrated lymphatic flow to the popliteal, sacral, and iliac lymph nodes in most mice and to the renal, inguinal, and lumbar-aortic lymph nodes in some mice. On the fusion images, the locations of the lymph nodes in the mouse trunk were in good agreement between FLI and MRI, indicating successful spatial registration even for the deep structures. The popliteal node tended to be visualized a little farther caudally in FLI than in MRI, presumably because the overlying tissues were thicker in the cranial portion. Integrated FLI/MRI lymphography with image fusion appears to be a useful tool for analysis of the murine lymphatic system.

Incidence of metastasis in circumflex iliac nodes distal to the external iliac nodes in intermediate- and high-risk endometrial cancer

Objective The aim of this study was to elucidate the incidence of metastasis in circumflex iliac nodes distal to the external iliac nodes (CINDEIN), which are also called suprainguinal nodes, in intermediate- and high-risk endometrial cancer. Removal of these nodes needs to be discussed from the viewpoint of patient's quality of life because removal of CINDEIN is strongly related to lower extremity lymphedema. Methods A retrospective chart review was carried out for 508 patients with intermediate- and high-risk endometrial cancer who were included in this study. We identified patients with lymph node metastasis. Lymph node sites were classified into four groups: (1) CINDEIN, (2) external iliac nodes, (3) Group A consisting of circumflex iliac nodes to the distal obturator nodes, internal iliac nodes, obturator nodes, cardinal ligament nodes (including deep obturator nodes), and sacral nodes, and (4) Group B consisting of common iliac nodes and para-aortic nodes. Logistic regression analysis was used to select risk factors for CINDEIN metastasis. Results In an analysis of 508 patients with intermediate- and high-risk disease, CINDEIN metastasis was found in fourteen (2.8%) of the patients. Multivariate analysis confirmed that high-risk histology (OR = 5.7, 95% CI = 1.2–16.1) and Group A node metastasis (OR = 9.7, 95% CI = 2.9–31.4) were independent risk factors for CINDEIN metastasis. None of the patients with G1 endometrioid adenocarcinoma had CINDEIN metastasis. Three (2.5%) of the patients with G2 endometrioid adenocarcinoma had CINDEIN metastasis and all of these three patients had other pelvic node metastasis. Conclusion Removal of CINDEIN can be eliminated in patients with G1 endometrial cancer and patients with G2 endometrial cancer who have no pelvic node metastasis.

Surgical Anatomy of the Retroperitoneal Spaces, Part III: Retroperitoneal Blood Vessels and Lymphatics

In this article, we discuss the surgical anatomy of the blood vessels and the lymphatic vessels and lymph nodes found in the retroperitoneum. Retroperitoneal blood vessels include the aorta and all its branches--parietal and visceral--from the diaphragm to the pelvis, and the inferior vena cava and its tributaries. The retroperitoneal lymphatics form a very rich and extensive chain. As a general rule, lymphatics follow the arteries and named lymph nodes are found at the root of the arteries. Retroperitoneal nodes of the abdomen comprise the inferior diaphragmatic nodes and the lumbar nodes. The latter are classified as left lumbar (aortic), intermediate (interaorticovenous), and right lumbar (caval). These nodes surround the aorta and the inferior vena cava. Around the aorta lie the paraortic nodes, preaortic nodes (include celiac, superior mesenteric, inferior mesenteric nodes collecting lymph from the splanchna supplied by the homonymous arteries), and retroaortic nodes. Similarly, around the vena cava lie the paracaval, precaval, and retrocaval nodes. Pelvic nodes include the common iliac, external and internal iliac, obturator, and sacral nodes.

Fluorescence Lymph Node Mapping in Living Mice Using Quantum Dots and a Compression Technique

The lymphatic system is essential in oncology and immunology, and in vivo fluorescence imaging plays a major role in assessing the lymphatic drainage. We investigated non-invasive fluorescence lymph node mapping in mice with special reference to the assessment of deep abdominal lymph nodes. Quantum dots were injected subcutaneously into the rear footpads of mice, and the time course of the fluorescent signal was assessed. Visualization of abdominal lymph nodes was compared with and without compression of the abdomen with transparent, colorless tape at injection doses of 1, 5, and 20 pmol. Popliteal, sacral, iliac, and renal lymph nodes were delineated by non-invasive imaging. Lymph node signals increased up to 3 h after injection. Compression of the abdomen markedly improved the visualization of the iliac nodes, which were invisible at 5 pmol without compression and visible at 1 pmol with compression. Fluorescence lymph node mapping using quantum dots allowed the visualization of deep abdominal lymph nodes in addition to superficial nodes in intact mice, with the aid of a simple compression technique.

Lymphoma‐Associated Polymyositis in Dogs

Lymphoma‐Associated Polymyositis in Dogs

Overexpression of vascular endothelial growth factor C increases growth and alters the metastatic pattern of orthotopic PC-3 prostate tumors

BackgroundProstate cancer metastasizes to regional lymph nodes and distant sites but the roles of lymphatic and hematogenous pathways in metastasis are not fully understood.MethodsWe studied the roles of VEGF-C and VEGFR3 in prostate cancer metastasis by blocking VEGFR3 using intravenous adenovirus-delivered VEGFR3-Ig fusion protein (VEGFR3-Ig) and by ectopic expression of VEGF-C in PC-3 prostate tumors in nude mice.ResultsVEGFR3-Ig decreased the density of lymphatic capillaries in orthotopic PC-3 tumors (p < 0.05) and inhibited metastasis to iliac and sacral lymph nodes. In addition, tumor volumes were smaller in the VEGFR3-Ig-treated group compared with the control group (p < 0.05). Transfection of PC-3 cells with the VEGF-C gene led to a high level of 29/31 kD VEGF-C expression in PC-3 cells. The size of orthotopic and subcutaneous PC-3/VEGF-C tumors was significantly greater than that of PC-3/mock tumors (both p < 0.001). Interestingly, while most orthotopic PC-3 and PC-3/mock tumors grown for 4 weeks metastasized to prostate-draining lymph nodes, orthotopic PC-3/VEGF-C tumors primarily metastasized to the lungs. PC-3/VEGF-C tumors showed highly angiogenic morphology with an increased density of blood capillaries compared with PC-3/mock tumors (p < 0.001).ConclusionThe data suggest that even though VEGF-C/VEGFR3 pathway is primarily required for lymphangiogenesis and lymphatic metastasis, an increased level of VEGF-C can also stimulate angiogenesis, which is associated with growth of orthotopic prostate tumors and a switch from a primary pattern of lymph node metastasis to an increased proportion of metastases at distant sites.

Dosimetric study of simplified intensity modulated radiation therapy for cervical cancer

Objective To compare the homogeneity and conformity of dose distribution in the target and the dose to the organs at risk among 3-dimensional conformal radiation therapy (3DCRT), intensity modulated radiation therapy (IMRT) and simplified intensity modulated radiation therapy (sIMRT), and then to evaluate the clinical value of sIMRT for cervical cancer. Methods Ten patients with cervical canc-er receiving radical radiotherapy and brachytherapy were enrolled. Before radiotherapy, CT was performed and target volumes were delineated. The clinical target volume (CTV) included supravaginal portion, cervi-cal stump, paracervical tissue, common iliac lymph nodes, internal and external iliac lymph nodes, obtura-tor lymph nodes, sacral lymph nodes, and the surrounding tissues. If the vagina was involved, the target vol-ume included the whole vagina. Margins between planning target volume (PTV) and CTV were 10 mm in the anterior direction, and 5 mm in other directions. The prescribed dose was 95% PTV receiving 45 Gy in 25 fractions of 1.8 Gy. The dose volume histogram, conformity index, homogeneity index and treatment time per faction were compared. Results Among the three radiotherapy techniques, 3DCRT had the best homo-geneity of dose distribution, while there was no significant difference between IMRT and slMRT. According to the conformity of dose distribution, sIMRT was better than 3DCRT, but worse than IMRT. Comparing the bladder sparing, IMRT was the best followed by sIMRT. The volume of the small intestine receiving high dose was increased significantly with slMRT when compared with IMRT. And the treatment time per faction was4, 10 and 18 minutes for3 DCRT, sIMRT and IMRT, respectively. Conclusions sIMRT could re-duce the work intensity of the staff, shorten the treatment time per fraction, and simplify the authenticate procedure, which is cost-effective and suitable in the treatment of cervical cancer. Key words: Cervical neoplasms/radiotherapy; Radiotherapy, simplified intensity-modulated; Dosimetry

Hyperosmotic stress contributes to mouse colonic inflammation through the methylation of protein phosphatase 2A

There are several reports suggesting hyperosmotic contents in the feces of patients suffering from inflammatory bowel disease (IBD). Previous works have documented that hyperosmolarity can cause inflammation attributable to methylation of the catalytic subunit of protein phosphatase 2A (PP2A) and subsequent NF-kappaB activation resulting in cytokine secretion. In this study, we demonstrate that dextran sulfate sodium (DSS) induces colitis due to hyperosmolarity and subsequent PP2A activation. Mice were randomized and fed with increased concentrations of DSS (0 mOsm, 175 mOsm, 300 mOsm, and 627 mOsm) for a duration of 3 wk or with hyperosmotic concentrations of DSS (627 mOsm) or mannitol (450 mOsm) for a duration of 12 wk. Long-term oral administration of hyposmotic DSS or mannitol had no demonstrable effect. Hyperosmotic DSS or mannitol produced a significant increase in colonic inflammation, as well as an increase in the weight of sacral lymph nodes and in serum amyloid A protein levels. Similar results were obtained through the ingestion of comparable osmolarities of mannitol. Hyperosmolarity induces the methylation of PP2A, nuclear p65 NF-kappaB activation. and cytokine secretion. The rectal instillation of okadaic acid, a well-known PP2A inhibitor, reverses the IBD. Short inhibiting RNAs (siRNAs) targeted toward PP2Ac reverse the effect of hyperosmotic DSS. The present study strongly suggests that DSS-induced chronic colitis is a consequence of the methylation of PP2Ac induced by hyperosmolarity.