Enhanced Lymph Node Research Articles

GMP-compliant sorting of dendritic cells for CCR7 expression improves therapeutic efficacy in cancer by enhancing lymph node migration and antigen-specific T cell activation

Background & Aim Dendritic cells (DCs) are a potentially powerful cell therapy due to their ability to induce T cell activation to defined antigens; however, this potential has not yet been realised in clinical trials. Monocyte-derived DCs (moDCs) are most commonly used in cancer therapy, although the rare blood DC subsets have also been explored. Crucially, cell function requires the chemokine receptor CCR7, which guides DCs from the site of antigen uptake to the lymph nodes. CCR7 expression, and therefore migration, is variable after isolation of DCs from blood or ex vivo culture, and may account for the limited efficacy seen in clinical trials. We hypothesised that selection of CCR7+ DCs by sorting would to improve cell function and therapeutic use of DCs. Methods, Results & Conclusion We have previously shown that murine CCR7+ bone marrow DCs (BMDCs) can be sorted using a novel chemokine-based system and show enhanced lymph node migration. This CCL19-fluorophore construct was shown to also detect expression of CCR7 on human DCs, and significantly enrich CCR7+ cells to high purity and viability (>90%) using magnetic bead and fluorescence-based sorting methods. Both murine and human CCR7+ DCs were more mature than CCR7- DCs by expression of CD80 and CD86 and production of T cell chemoattractants such as CCL17 and CCL22. In vitro, CCR7+ DCs were also most effective in stimulating antigen-specific T cells, especially memory T cells producing IFNγ, TNFα and IL-2. In vivo, sorted murine CCR7+ BMDCs, but not unsorted DCs, were sufficient to reduce subcutaneous and metastatic tumour growth in the B16F10.ova mouse model. Increased migration of injected DCs to lymph nodes led to an elevated overall number of tumour-specific T cells in CCR7+ DC-receiving mice, but this was not seen in those receiving unsorted DCs. In agreement with in vitro data, CCR7+ DCs increased production of mature tumour-specific T cells, particularly with the desirable memory (CD62L+CD44+) phenotype, suggesting their potency in a human therapeutic context. As production of this chemokine construct is possible at GMP grade, this protocol is amenable to GMP translation using the MACSQuant Tyto cell sorter. Purity and viability of CCR7+ DCs remained consistently high between the original and GMP-ised protocols, with an improved cell yield versus bead sorting. Overall, this novel sorting process is compliant with current GMP manufacture and creates a realistic DC product with enhanced function and potential therapeutic benefit.

NDRG4 promoter hypermethylation is a mechanistic biomarker associated with metastatic progression in breast cancer patients

The risk of developing metastatic disease in breast cancer patients is traditionally predictable based on the number of positive axillary lymph nodes, complemented with additional clinicopathological factors. However, since lymph node-negative patients have a 20–30% probability of developing metastatic disease, lymph node information alone is insufficient to accurately assess individual risk. Molecular approaches, such as multigene expression panels, analyze a set of cancer-related genes that more accurately predict the early risk of metastasis and the treatment response. Here, we present N-Myc downstream-regulated gene 4 (NDRG4) epigenetic silencing as a mechanistic biomarker of metastasis in ductal invasive breast tumors. While aberrant NDRG4 DNA hypermethylation is significantly associated with the development of metastatic disease, downregulation of NDRG4 transcription and protein expression is functionally associated with enhanced lymph node adhesion and cell mobility. Here, we show that epigenetic silencing of NDRG4 modulates integrin signaling by assembling β1-integrins into large punctate clusters at the leading edge of tumor cells to promote an “adhesive switch,” decreasing cell adhesion to fibronectin and increasing cell adhesion and migration towards vitronectin, an important component of human lymph nodes. Taken together, our functional and clinical observations suggest that NDRG4 is a potential mechanistic biomarker in breast cancer that is functionally associated with metastatic disease.

Idiopathic Granulomatous Mastitis - a new approach in diagnostics and treatment.

Idiopathic granulomatous mastitis (IGM) is a rare inflammatory breast disease mimicking carcinoma and puerperal or non-puerperal mastitis. The primary purpose of this prospectively performed case control study was to compare clinical and imaging signs of IGM with the reference group of nonspecific, non- puerperal mastitis (NM) to identify the most typical clinical and imaging signs essential for a correct differential diagnosis. The secondary purpose was to present a new approach to non-invasive treatment. Thirty-nine women with histologically proven IGM and twenty-six patients with nonspecific mastitis underwent clinical examination, breast ultrasound (US), mammography (MG) and MRI examination. The most typical signs were selected for each group, and method and were statistically evaluated. The effectivity of colchicine, vitamin E and ribwort plantain tincture in treatment was assessed by clinical examination and imaging. Typical clinical signs of IGM included unilateral acute onset of breast edema, redness, palpable masses, missing fever, lymphadenopathy, no response to antibiotics or surgical interventions. Ultrasound revealed: "finger-like" structures (100%), ductectasias (76.9%), abscesses (76.9%), and lymphadenopathy (15.4%), while in MRI skin and tissue edema (100%), multicentric lesions (100%), abscesses (76.9%), ring enhancement (84.6%), lymphadenopathy (15.4%) and small enhancing lymph nodes (38.5%) were observed. Among the clinical signs, fistulas, hypoechoic mass, ductectasias and diffusion weighted images (DWI) restriction were significantly more frequent in patients with IGM than in those with NM. Treatment effectivity yielded 100% with a complete response between 6-19 months, depending on the disease extent. Targeted questions together with imaging can speed up selection for proper treatment with colchicine, vitamin E and local treatment. Long lasting use of antibiotics and repeated surgical interventions should be avoided.

MTORC2 deficiency in cutaneous dendritic cells potentiates CD8+ effector T cell responses and accelerates skin graft rejection.

Mechanistic target of rapamycin (mTOR) complex (mTORC)1 and mTORC2 regulate the differentiation and function of immune cells. While inhibition of mTORC1 antagonizes dendritic cell (DC) differentiation and suppresses graft rejection, the role of mTORC2 in DCs in determining host responses to transplanted tissue remains undefined. Using a mouse model in which mTORC2 was deleted specifically in CD11c+ DCs (TORC2DC-/- ), we show that the transplant of minor histocompatibility Ag (HY)-mismatched skin grafts from TORC2DC-/- donors into wild-type recipients results in accelerated rejection characterized by enhanced CD8+ T cell responses in the graft and regional lymphoid tissue [Correction added on January 9, 2019, after first online publication: in the previous sentence, major was changed to minor]. Similar enhancement of CD8+ effector T cell responses was observed in MHC-mismatched recipients of TORC2DC-/- grafts. Augmented CD8+ T cell responses were also observed in a delayed-type hypersensitivity model in which mTORC2 was absent in cutaneous DCs. These elevated responses could be ascribed to an increased T cell stimulatory phenotype of TORC2DC-/- and not to enhanced lymph node homing of the cells. In contrast, rejection of ovalbumin transgenic skin grafts in TORC2DC-/- recipients was unaffected. These findings suggest that mTORC2 in skin DCs restrains effector CD8+ T cell responses and have implications for understanding of the influence of mTOR inhibitors that target mTORC2 in transplant.

Platelet deficiency in Tpo-/- mice can both promote and suppress the metastasis of experimental breast tumors in an organ-specific manner.

Platelets are thought to play an important role in metastasis formation, although the mechanisms involved remain incompletely understood. Here we studied the influence of platelet numbers on organ-specific metastasis to the lungs and lymph nodes using Tpo deficient mice that have low platelet counts. After tail vein injection of 4T1 breast cancer cells, the number of lung metastases was significantly lower in Tpo-/- mice compared to Tpo+/+ mice. The same was true for the bone-tropic 4T1.2 derivative. In spontaneous orthotopic metastasis assays, 4T1 and 4T1.2 primary tumor growth was not affected by the genotype of the mice. However, the number of 4T1.2 lung metastases was significantly lower in Tpo-/- mice compared to Tpo+/+ mice, whereas the number of 4T1 lung metastases was unaffected. Moreover, in mice bearing 4T1 tumors, lymph node metastases were larger in the Tpo-/- background, and lymph node metastasis frequency was higher in Tpo-/- mice bearing 4T1.2 tumors compared to that in wild-type mice. Enhanced lymph node metastasis in Tpo-/- mice was not associated with changes in peritumoral lymphatic vessel density in the primary tumors. Together, our data indicate that platelets do not affect primary tumor growth in this breast cancer model, but can differentially influence site-specific metastasis to lymph nodes and lungs.

Size-Controlled Iron Oxide Nanoplatforms with Lipidoid-Stabilized Shells for Efficient Magnetic Resonance Imaging-Trackable Lymph Node Targeting and High-Capacity Biomolecule Display.

Nanoplatforms for biomolecule delivery to the lymph nodes have attracted considerable interest as vectors for immunotherapy. Core-shell iron oxide nanoparticles are particularly appealing because of their potential as theranostic magnetic resonance imaging (MRI)-trackable vehicles for biomolecule delivery. The key challenge for utilizing iron oxide nanoparticles in this capacity is control of their coating shells to produce particles with predictable size. Size determines both the carrier capacity for biomolecule display and the carrier ability to target the lymph nodes. In this study, we develop a novel coating method to produce core-shell iron oxide nanoparticles with controlled size. We utilize lipidlike molecules to stabilize self-assembled lipid shells on the surface of iron oxide nanocrystals, allowing the formation of consistent coatings on nanocrystals of varying size (10-40 nm). We further demonstrate the feasibility of leveraging the ensuing control of nanocarrier size for optimizing the carrier functionalities. Coated nanoparticles with 10 and 30 nm cores supported biomolecule display at 10-fold and 200-fold higher capacities than previously reported iron oxide nanoparticles, while preserving monodisperse sub-100 nm size populations. In addition, accumulation of the coated nanoparticles in the lymph nodes could be tracked by MRI and at 1 h post injection demonstrated significantly enhanced lymph node targeting. Notably, lymph node targeting was 9-40 folds higher than that for previously reported nanocarriers, likely due to the ability of these nanoparticles to robustly maintain their sub-100 nm size in vivo. This approach can be broadly applicable for rational design of theranostic nanoplatforms for image-monitored immunotherapy.

Preoperative sentinel lymph node localization in vulvar cancer: preliminary experience with inguinal intradermal contrast-enhanced ultrasound.

To evaluate the feasibility and diagnostic performance of intradermal contrast-enhanced ultrasound (CEUS) sentinel lymph node (SLN) procedure in vulvar cancer. Twelve consecutive patients with vulvar cancer underwent preoperatively inguinal CEUS SLN examination and guide wire marking of the enhanced lymph nodes. Altogether, 20 groins were examined with CEUS contrast agent injections including 8 bilateral groins due to midline tumours. One groin was excluded due to previous inguinal surgery. The results of the CEUS examinations were compared to conventional SLN biopsy using radiocolloid scintigraphy and/or methylene blue dye and final postoperative histopathology. The inguinal sentinel CEUS procedure had a technical success rate of 94.7% (18/19 injections) for identifying a potential inguinal SLN. Conventional SLN biopsy using lymphoscintigraphy and/or methylene blue dye was successfully performed in 16 groins. Compared to conventional SLN biopsy, the overall sensitivity was 81.2% (13/16 injections). Additionally, CEUS detected enhancing SLNs in two cases when traditional SLN procedure failed to do so. All metastatic SLNs (n = 5) were correctly identified by CEUS procedure. Intradermal CEUS SLN localization can be applied in the inguinal lymphatic region in patients with vulvar cancer. Further studies are needed to verify the clinical value of this method. • CEUS is a feasible method for inguinal SLN detection in vulvar cancer • All metastatic inguinal SLNs were identified by CEUS procedure • Further studies are needed to verify the clinical value of this method.

Engineering intranasal mRNA vaccines to enhance lymph node trafficking and immune responses.

As we face outbreaks of pandemic diseases such as Zika virus, intranasal mRNA vaccination provides instant massive protection against highly variant viruses. Various polymer-based delivery systems have been successfully applied in intranasal vaccine delivery. However, the influence of molecular structure of the polymeric carriers on the lymph node trafficking and dendritic cell maturation is seldom studied for intranasal vaccination. Therefore, engineering polymer-based vaccine delivery system and elucidating the relationship between molecular structure and the intranasal delivery efficiency are essential for maximizing the immune responses. We hereby construct self-adjuvanting polymer-based intranasal mRNA vaccines to enhance lymph node trafficking and further improve immune responses.

Combinatorial Delivery of Dual and Triple TLR Agonists via Polymeric Pathogen-like Particles Synergistically Enhances Innate and Adaptive Immune Responses

Despite decades of research very few vaccine-adjuvants have received FDA approval. Two fundamental challenges plague clinical translation of vaccine-adjuvants: reducing acute toxicities that result from systemic diffusion of many soluble adjuvants, and delivering multiple adjuvants at the same time to mimic the synergistic immune-stimulation of pathogens, while being safe. In order to address these barriers, we evaluated combinations of four clinically relevant immune-agonists, specifically Toll-like receptor (TLR) ligands, using biodegradable, polymer microparticles. We tested them alone and in combinations of 2 or 3, for a total of 10 unique conditions. We evaluated primary bone-marrow-derived Dendritic Cell phenotypes and functionality, and identified several synergistic combinations. We picked a dual and a triple adjuvant combination, TLR4/TLR9 and TLR4/TLR7/TLR9, for further evaluation and found that both combinations promoted antigen cross-presentation in vitro. Studies in mice using the model antigen Ovalbumin, showed that both combinations enhanced lymph node germinal center and T follicular helper cell responses. The triple adjuvant combination showed increased antigen-specific antibody titer with an overall balanced Th1/Th2 response, while the dual combination promoted Th1-polarized IgG responses. Our results show how polymeric particulate-carriers can be adopted to safely deliver combinatorial adjuvants and selectively synergize specific types of immune responses for vaccine applications.

Display of DNA on Nanoparticles for Targeting Antigen Presenting Cells.

Efficient delivery of antigens is of paramount concern in immunotherapies. We aimed to target antigen presenting cells (APCs) by conjugating CpG oligonucleotides to an E2 protein nanoparticle surface (CpG-PEG-E2). Compared to E2 alone, we observed ~4-fold increase of in vitro APC uptake of both CpG-PEG-E2 and E2 conjugated to non-CpG DNA. Furthermore, compared to E2-alone or E2 functionalized solely with polyethylene glycol (PEG), the CpG-PEG-E2 showed enhanced lymph node retention up to at least 48 hr and 2-fold increase in APC uptake in vivo, parameters which are advantageous for vaccine success. This suggests that enhanced APC uptake of nanoparticles mediated by oligonucleotide display may help overcome delivery barriers in vaccine development.

Upregulation of Serum miR-10b Is Associated with Poor Prognosis in Patients with Melanoma.

Aberrant expression of microRNAs (miRNAs) are believed to play a central role in the initiation and development of cancer. The aim of our study was to determine the clinical significance of serum miR-10b in melanoma. A total of 85 and 30 serum samples were obtained from patients with melanoma and healthy volunteers respectively. qRT-PCR was performed to evaluate the expression level of miR-10b in the melanoma cell lines and the serum samples from the participants. Then the clinical significance of serum miR-10b was further determined. Our results showed that the expression level of miR-10b was significantly increased in metastasis melanoma cells and melanoma patients compared to their respective controls. In addition, serum miR-10b expression level was able to discriminate melanoma patients from healthy volunteers as well differentiate melanoma patients at different clinical stage with high accuracy. Moreover, upregulation of serum miR-10b was positively associated with enhanced lymph node metastasis, advanced clinical stage and a shortened survival rate. Finally serum miR-10b was an independent prognostic factor for melanoma. Collectively, our study suggests that serum miR-10b level is upregulated in melanoma and associated with poor prognosis. It may be used as a potential prognostic biomarker for melanoma.

Plant viral nanoparticles-based HER2 vaccine: Immune response influenced by differential transport, localization and cellular interactions of particulate carriers

Cancer vaccines are designed to elicit an endogenous adaptive immune response that can successfully recognize and eliminate residual or recurring tumors. Such approaches can potentially overcome shortcomings of passive immunotherapies by generating long-lived therapeutic effects and immune memory while limiting systemic toxicities. A critical determinant of vaccine efficacy is efficient transport and delivery of tumor-associated antigens to professional antigen presenting cells (APCs). Plant viral nanoparticles (VNPs) with natural tropism for APCs and a high payload carrying capacity may be particularly effective vaccine carriers. The applicability of VNP platform technologies is governed by stringent structure-function relationships. We compare two distinct VNP platforms: icosahedral cowpea mosaic virus (CPMV) and filamentous potato virus X (PVX). Specifically, we evaluate invivo capabilities of engineered VNPs delivering human epidermal growth factor receptor 2 (HER2) epitopes for therapy and prophylaxis of HER2+ malignancies. Our results corroborate the structure-function relationship where icosahedral CPMV particles showed significantly enhanced lymph node transport and retention, and greater uptake by/activation of APCs compared to filamentous PVX particles. These enhanced immune cell interactions and transport properties resulted in elevated HER2-specific antibody titers raised by CPMV- vs. PVX-based peptide vaccine. The 'synthetic virology' field is rapidly expanding with numerous platforms undergoing development and preclinical testing; our studies highlight the need for systematic studies to define rules guiding the design and rational choice of platform, in the context of peptide-vaccine display technologies.

USPIO enhanced lymph node MRI using 3D multi-echo GRE in a rabbit model.

Ultrasmallsuperparamagnetic iron oxide (USPIO) has been suggested to be a negative MR contrast agent to detect metastatic lymph nodes. Previously reported studies have evaluated the diagnostic performance of USPIO-enhanced MR lymph node imaging based on signal intensity. In this study, we investigate the specific performance of three different parametric approaches (normalized signal intensity, R2 * and susceptibility) using 3D multi-echo gradient echo to quantify the USPIO particles in lymph nodes. Nine rabbits with VX2 tumor implants were scanned before and after USPIO injection. From 3D multi-echo GRE magnitude and phase data, we generated multi-echo combined T2 *-weighted images, an R2 * map, and a quantitative susceptibility map. Eighteen lymph nodes (nine reactive and nine metastatic) were evaluated and showed remarkable signal drops in the area of USPIO accumulation. On parametric analysis, the R2 * difference before and after USPIO injection was significantly different (p < 0.05) between reactive and metastatic lymph nodes; in contrast, the normalized signal intensity and susceptibility were not significantly different between the nodes. Our study showed the potential utility of USPIO-enhanced MRI using R2* mapping from 3D multi-echo GRE for the detection of lymph node metastasis and parametric analysis of lymph node status in a rabbit model. Copyright © 2016 John Wiley & Sons, Ltd.

Methylation of CDX2 as a Predictor in Poor Clinical Outcome of Patients with Colorectal Cancer

Increased DNA methylation within the promoter regions of tumor suppressor genes has been associated with gene silencing in various cancers. CDX2, a caudal-related homeobox transcription factor, represents a key tumor suppressor in colorectal cancer. However, the pathological role of its promoter methylation has not yet been well defined in colorectal cancer. We measured CDX2 promoter methylation by methylation-specific PCR (MSP) and CDX2 mRNA levels by real-time quantitative reverse transcription-PCR in both tumor and normal control tissues collected from 108 colorectal cancer patients and then performed correlation analyses. We also determined whether CDX2 promoter methylation status was associated with any clinicopathological features and prognosis in these patients. CDX2 was expressed in all normal mucosa, but only in 38 (35.2%) tumor tissues. CDX2 promoter methylation was detected in 43 (39.8%) tumor samples, but in none of the normal tissue specimens. DNA methylation correlated with decreased mRNA level in these tumors (p < 0.05). The methylation of the CDX2 promoter was associated with enhanced lymph node metastases and shorter survival time (p = 0.0273), but was independent of tumor stage, tumor differentiation, gender, and age. Enhanced CDX2 promoter methylation is associated with gene silencing in a subgroup of colorectal cancer patients with lymph node metastasis and shorter survival times.

Altered Sensorium and ST Segment Elevation: Atypical Manifestations of Hypercalcemia in a Renal Transplant Patient With Mycobacterium Avium Intracellulare (MAI)

SESSION TITLE: Student/Resident Case Report Poster - Critical Care I SESSION TYPE: Student/Resident Case Report Poster PRESENTED ON: Tuesday, October 25, 2016 at 01:30 PM - 02:30 PM INTRODUCTION: Hypercalcemia secondary to granulomatous diseases is a known cause of hypercalcemia. This case describes hypercalcemia secondary to MAI in a renal transplant patient treated emergently with hemodialysis with resolution of symptoms and calcium. ST segment elevation in inferior leads was also noted on electrocardiogram(EKG). CASE PRESENTATION: A 75 year old woman presented to the ED with altered sensorium and slurred speech, which developed after a hemodialysis treatment. On examination, she was disoriented and noted to have matted cervical lymphadenopathy. Lab results showed a corrected calcium of 13.74 mg/dl. Cardiac enzymes were negative and 12 lead EKG(Fig-1) revealed ST segment elevation in leads II, III and aVF. The patient underwent emergent dialysis, calcitonin was administered with correction of hypercalcemia. PTH was 38.5pg/ml, TSH was 2.27 U/ml, Vitamin D2 was <8ng/ml, vitamin D3 was 91 ng/ml and Alkaline Phosphatase 159 IU/L. CT Neck w/ IV contrast (Fig-2) demonstrated large cystic/necrotic enhancing lymph nodes. Cultures from a draining lymph node grew multiple acid fast bacilli, which on further microbiological analysis (Fig-3) revealed MAI. The patient was treated with Azithromycin and Rifabutin. Pamidronate was continued once weekly. The patient’s EKG also normalized. DISCUSSION: Hypercalcemia is a metabolic emergency noted in approximately 0.5% hospitalized patients1. The principal mechanism of hypercalcemia in granulomatous disease has been attributed to uninhibited production of 1,25 (OH)2 Vitamin D from extra-renal sources. Alveolar macrophages independently produce 1 a hydroxylase, leading to increased 1,25 (OH)2 Vitamin D and low vitamin D1. Glucocorticoids, chloroquine, ketoconazole and bisphosphonates have been used to treat hypercalcemia secondary to MAI, however some patients required hemodialysis. ST segment elevation is a rare finding of hypercalcemia found on electrocardiography. Disseminated MAI infection leading to hypercalcemia is rare. CONCLUSIONS: To our knowledge hypercalcemia is a rare complication resulting from disseminated MAI infection. Our patient also had atypical findings of ST segment elevation on EKG warranting the need for close cardiac monitoring in such patients. Reference #1: Roussos, A, Lagogianni, Gonis, A, Ilias I, Kazi, D, Patsopoulos, Philippou, N. Hypercalcemia in Greek patients with tuberculosis before the initiation of anti-tuberculosis treatment. Respiratory Medicine, 2001;95,187-190. Reference #2: Sharma, OP. Hypercalcemia in granulomatous disorders: a clinical review. Current Opinion in Pulmonary Medicine 2000, 6:442-447. Reference #3: L. Littmann, L. Taylor, and W. D. Brearley, ST-segment elevation: a common finding in severe hypercalcemia. Journal of Electrocardiology, vol. 40, no. 1, pp. 60-62, 2007. DISCLOSURE: The following authors have nothing to disclose: Shyam Shankar, Karan Wats, Hitesh Raheja, Ishan Malhotra, Nupur Uppal, Mangalore Amith Shenoy, Stephan Kamholz No Product/Research Disclosure Information

Migration of dendritic cells to the lymph nodes and its enhancement to drive anti-tumor responses.

Better prognoses associated with increased T cell infiltration of tumors, as seen with chimeric antigen receptor (CAR) T cell therapies and immune checkpoint inhibitors, portray the importance and potential of the immune system in controlling tumors. This has rejuvenated the field of cancer immunotherapy leading to an increasing number of immunotherapies developed for cancer patients. Dendritic Cells (DCs) vaccines represent an appealing option for cancer immunotherapy since DCs have the ability to circumvent tolerance to tumors by its adjuvant properties and to induce memory T cells that can become persistent after initial tumor clearance to engage potential metastatic tumors. In the past, DC-based cancer vaccines have elicited only poor clinical response in cancer patients, which can be attributed to complex and a multitude of issues associated with generation, implementing, delivery of DC vaccine and their potential interaction with effector cells. The current review mainly focuses on migration/trafficking of DCs, as one of the key issues that affect the success of DC-based cancer vaccines, and discusses strategies to enhance it for cancer immunotherapy. Additionally, impact of maturation, route of DC delivery and negative effects of tumor microenvironment (TME) on DC homing to LN are reviewed. Moreover, strategies to increase the expression of genes involved in Lymph node homing, preconditioning of the vaccination site, enhancing lymph node ability to attract and receive DCs, while limiting negative impact of TME on DC migration are discussed.

Castleman Disease of the Thorax: Clinical, Radiologic, and Pathologic Correlation: From the Radiologic Pathology Archives.

Castleman disease is a complex lymphoproliferative disease pathologically divided into two subtypes, the hyaline vascular variant (HVV) and the plasma cell variant (PCV). The HVV is the most common, is thought to represent a benign neoplasm of lymph node stromal cells, and is treated with surgical resection. It is most commonly found in the mediastinum, where it classically appears as a unicentric, avidly enhancing mass at computed tomography (CT) and magnetic resonance imaging. This appearance can mimic other avidly enhancing mediastinal masses, and location, clinical history, laboratory data, and nuclear medicine single photon emission CT (SPECT) and positron emission tomography (PET) studies can help narrow the differential diagnosis. Multicentric Castleman disease (MCD), which in the majority of cases is composed of the PCV, is an aggressive lymphoproliferative disorder associated with human herpesvirus infection, interleukin 6 dysregulation, and other systemic disorders. While it can be difficult to differentiate MCD from lymphoma, the presence of avidly enhancing lymph nodes can suggest the diagnosis. The purpose of this article is to review the clinical, immunologic, and pathologic findings associated with both unicentric Castleman disease and MCD and discuss how the imaging findings correlate with the pathophysiology of the disease.

Clinical application of carbon nanoparticles lymphatic tracer technology to guide surgery for gallbladder cancer

Objective To study carbon nanoparticles lymphatic tracer to guide surgery for gallbladder cancer. Methods 120 patients with gallbladder cancer were randomized into two groups: the experimental group (n=60) and the control group (n=60). For the experimental group, 0.1 ml carbon nanoparticles was injected at 4~6 locations subserosally around the cancerous site intraoperatively. Stained lymph nodes were used as markers to guide lymphadenectomy. The resected specimens were carefully dissected, and then the lymph nodes were studied according to their positions by histopathological examinations. Results Various degrees of stained lymph nodes were seen in the specimens. In the experimental group, the number of lymph nodes (12.0±3.8), and metastatic lymph nodes (6.3±3.3) per patient were obviously higher than those in the control group (9.2±3.6、4.4±2.8), respectively, (P<0.05). There were significant differences (P<0.05) in postoperative disease-free survival and overall survival. Conclusions Intraoperative injection of carbon nanoparticles enhanced lymph node clearance and increased the number of lymph nodes and metastatic lymph nodes dissected, which helped to guide pathological staging. There were significant improvements in disease-free survival and overall survival of these patients with gallbladder cancer after surgery. Key words: Nano carbon; Lymphatic tracer technology; Gallbladder cancer; Lymphadenectomy

Up- regulation of miR-328-3p sensitizes non-small cell lung cancer to radiotherapy.

MicroRNAs (miRNAs) are believed to be resistant against radiotherapy in certain types of cancers. The aim of our study was to determine the clinical application of miRNAs in non-small cell lung cancer (NSCLC). Sixty NSCLC tissue samples and adjacent histologically normal tissues were obtained for miRNAs microarray analysis and validated by RT-qPCR. Correlation between miRNA expression level and clinicopathological features was evaluated. Our study examined the influence of changed miRNA expression on the damaged DNA and its associated radio sensitivity. Luciferase assay was performed to determine potential effects on the targeted gene. Our study identified fifteen altered miRNAs in which miR-328-3p was down regulated in NSCLC tumour tissue as compared to normal tissues. Down-expression of miR-328-3p was positively associated with an enhanced lymph node metastasis, advanced clinical stage and a shortened survival rate. miR-328-3p expression was decreased in A549 cells compared to other NSCLC cell lines. Up-regulation of miR-328-3p demonstrated a survival inhibition effect in A549 and restored NSCLC cells’ sensitivity to radio therapy. An increased miR-328-3p expression promoted irradiation-induced DNA damage in cells. γ-H2AX was identified as the direct target of miR-328-3p. Over-expressed miR-328-3p can improve the radiosensitvity of cells by altering the DNA damage/repair signalling pathways in NSCLC.

Value of Computerized Tomography Enterography in Predicting Crohn's Disease Activity: Correlation with Crohn's Disease Activity Index and C-Reactive Protein.

BackgroundThe accurate evaluation of Crohn’s disease activity is important for the treatment of the disease and for monitoring the response. Computerized tomography (CT) enterography is a useful imaging modality that reflects enteric inflammation, as well as extramural complications.ObjectivesThe aim of this study was to evaluate the correlation between CT enterographic (CTE) findings of active Crohn’s disease and the Crohn’s disease activity index (CDAI) and C-reactive protein (CRP).Patients and MethodsFifty CT enterographies of 39 patients with Crohn’s disease in the small bowel were used in our study. The CDAI was assessed through clinical and laboratory variables. Multiple CT parameters, including mural hyperenhancement, mural thickness, mural stratification, comb sign, and mesenteric fat attenuation, were evaluated with a four-point scale. The presence or absence of enhanced lymph nodes, fibrofatty proliferation, sinus or fistula, abscess, and stricture were also assessed. Two gastrointestinal radiologists independently reviewed all CT images, and inter-observer agreement was examined. Correlations between CT findings, CRP, and CDAI were assessed using Spearman’s rank correlation and logistic regression analysis. To assess the predictive accuracy of the model, a receiver-operating characteristic curve analysis for the sum of CT enterographic scores was used.ResultsMural hyperenhancement, mural thickness, comb sign, mesenteric fat density, and fibrofatty proliferation were significantly correlated with CDAI and CRP (P < 0.05). The binary logistic regression model demonstrated that mesenteric fat density, mural stratification, and the presence of enhanced lymph nodes (P < 0.05) had an influence on CDAI severity. The area under the receiver operating characteristic curve (AUROC) of the CTE index for predicting disease activity was 0.85. Using a cut-off value of 8, the sensitivity and negative predictive values were 95% and 94%, respectively.ConclusionMost CTE findings correlated with CDAI and CRP in patients with active Crohn’s disease.