Primary Lymph Node Research Articles

Deciphering head and neck cancer microenvironment: Single-cell and spatial transcriptomics reveals human papillomavirus-associated differences.

Human papillomavirus (HPV) is a major causative factor of head and neck squamous cell carcinoma (HNSCC), and the incidence of HPV- associated HNSCC is increasing. The role of tumor microenvironment in viral infection and metastasis needs to be explored further. We studied the molecular characteristics of primary tumors (PTs) and lymph node metastatic tumors (LNMTs) by stratifying them based on their HPV status. Eight samples for single-cell RNA profiling and six samples for spatial transcriptomics (ST), composed of matched primary tumors (PT) and lymph node metastases (LNMT), were collected from both HPV- negative (HPV- ) and HPV-positive (HPV+ ) patients. Using the 10x Genomics Visium platform, integrative analyses with single-cell RNA sequencing were performed. Intracellular and intercellular alterations were analyzed, and the findings were confirmed using experimental validation and publicly available data set. The HPV+ tissues were composed of a substantial amount of lymphoid cells regardless of the presence or absence of metastasis, whereas the HPV- tissue exhibited remarkable changes in the number of macrophages and plasma cells, particularly in the LNMT. From both single-cell RNA and ST data set, we discovered a central gene, pyruvate kinase muscle isoform 1/2 (PKM2), which is closely associated with the stemness of cancer stem cell-like populations in LNMT of HPV- tissue. The consistent expression was observed in HPV- HNSCC cell line and the knockdown of PKM2 weakened spheroid formation ability. Furthermore, we found an ectopic lymphoid structure morphology and clinical effects of the structure in ST slide of the HPV+ patients and verified their presence in tumor tissue using immunohistochemistry. Finally, the ephrin-A (EPHA2) pathway was detected as important signals in angiogenesis for HPV- patients from single-cell RNA and ST profiles, and knockdown of EPHA2 declined the cell migration. Our study described the distinct cellular composition and molecular alterations in primary and metastatic sites in HNSCC patients based on their HPV status. These results provide insights into HNSCC biology in the context of HPV infection and its potential clinical implications.

Radiation induced brachial plexopathy in head and neck cancer patients treated with definitive radiotherapy and correlation with disease characteristics and dosimetric parameters.

Definitive concurrent chemoradiotherapy (CRT) is the standard of care in advanced stages of head and neck cancer (HNC). With evident increase in survival rate there is also simultaneous increase in toxicity affecting the quality of life. One of the less researched late toxicity is radiation induced brachial plexopathy (RIBP). In this dosimetric study we intent to contour the brachial plexus (BP) as an organ at risk (OAR) and determine the factors that contribute to dose variations to BP, and clinically evaluate the patients for RIBP during follow-up using a questionnaire. 30 patients with HNC planned for CRT from September 2020 to June 2022 were accrued. Patients were treated to a dose of 6600 cGy with intensity modulated radiotherapy using the simultaneous integrated boost technique. From the dose-volume histogram (DVH) statistics the BP volume, Dmax and other parameters like V66, V60 were assessed and was correlated with respect to primary tumour and nodal stage. On corelation, more than the T stage, the N stage and the primary location had a significant impact on the Dmax. With a median follow-up of 17.9 months, the incidence of RIBP was 6.67%. The 2-year disease free survival and the 2-year overall survival were 53.7% and 59.4%, respectively. In oropharyngeal/hypopharyngeal primaries and in advanced nodal disease, BP receives higher doses contributing to RIBP. Primary tumor and nodal stage also impacted V60 and V66 of BP. Hence, contouring of BP as an OAR becomes imperative, and respecting the DVH parameters is essential.

Incidental Primary Cystic Duct Lymph Node Tuberculosis in a 28-year-old Female: A Rare Case Report

Tuberculosis (TB) is an infectious disease caused by acid-fast bacilli Mycobacterium tuberculosis (MTB). The lung remains the most common site involved by it. Extrapulmonary sites of TB are lymph nodes, pleura, bone, joints, urogenital tract, and meninges. Isolated Cystic Duct Tubercular Lymphadenitis (CDL TB) without involving the gallbladder is an uncommon manifestation of TB that primarily affects the lymph nodes around the cystic duct. A multidisciplinary approach is essential for accurate diagnosis and successful management of such cases. The authors hereby present a case of a 28-year-old female, without any evidence of TB elsewhere and an incidental finding of CDL TB. “The patient was sent with a clinical diagnosis of Chronic Calculus Cholecystitis (CCC)”. This highlights the need for comprehensive clinical evaluation and diagnostic investigations to avoid misdiagnosis or delayed management. This underscores the significance of maintaining a high index of suspicion for extrapulmonary TB, particularly in regions where the disease remains endemic. Healthcare practitioners should be vigilant when encountering unusual clinical presentations or when dealing with atypical combinations of gallbladder pathologies, as illustrated by the concurrent occurrence of CDL TB with CC with evidence of Pyloric Metaplasia (PM) and cholesterolosis without cholelithiasis, as in the patient of the present case report.

En Bloc Resection of a Primary Tumor and Lymph Nodes in Non-Small-Cell Lung Cancer.

We established a novel surgical procedure for resectable non-small-cell lung cancer (NSCLC), which involves resection of the affected lobe and regional lymph nodes without separation, namely en bloc surgery. We introduced the technical details and early and late outcomes by comparing them with those of conventional surgery. We retrospectively analyzed patients who underwent lobectomy with hilar and mediastinal lymph node dissection for stages I-III NSCLC. A propensity score-matched analysis was performed based on demographic variables. Propensity score-matching yielded 317 pairs. En bloc surgery was not associated with a longer operation time, a higher amount of intraoperative bleeding, or a higher frequency of postoperative complications. The number of resected lymph nodes (P = 0.277) and frequency of N upstaging (P = 0.587) did not differ between the groups. However, en bloc surgery was associated with higher overall survival in comparison to conventional surgery (P = 0.012). According to a stratification analysis, the survival advantage of en bloc surgery over conventional surgery was remarkable in pathological N-positive disease (P = 0.005), whereas it disappeared in pathological N-negative disease (P = 0.147). En bloc surgery is feasible and can be performed in patients with possible N-positive NSCLC.

NEOADJUVANT CHEMORADIOTHERAPY IN THE MULTIMODALITY TREATMENT OF STAGE IIIA(N2) NON-SMALL CELL LUNG CANCER (NSCLC)

Summary. The choice of optimal treatment for stage IIIA(N2) non-small cell lung cancer (NSCLC) remains controversial, since the five-year survival rate of this category of patients cannot meet modern medical requirements and patient expectations even after radical surgery and chemotherapy (CT). Accordingly, the analysis of available clinical data and the search for the most effective multimodal treatment option are constantly ongoing. The main treatment strategy for stage IIIA(N2) NSCLC is surgery with induction CT. As part of a comprehensive clinical approach, neoadjuvant CT (NCT) is used to eliminate distant micrometastases, leading to increased survival in an independent operation. Meanwhile, induction with concurrent chemoradiotherapy (CRT) followed by surgery resulted in a 30% to 40% improvement in 5-year survival. Aim: to evaluate the results of treatment of patients with stage IIIA(N2) NSCLC after the use of various methods of neoadjuvant and adjuvant therapy in case of surgical intervention. Object and methods: the effect of NCRT was performed in 37 patients with stage IIIA(N2) NSCLC in selected tumors of the lungs and the middle of the National Cancer Institute. This group of patients was planned for multimodal treatment, which included 3 or 4 cycles of chemotherapy with platinum-based drugs following the regimens of cisplatin + docetaxel or carboplatin + paclitaxel with one-hour radiotherapy (RT), consisting of sessions in the or classical fractionation with a single dose of 2 Gy, SOD up to 50 Gy with subsequent surgery. The number of CT cycles (3 or 4) depended on the result of the control observation of the dynamics of regression of the tumor process. In the core of the vicoristan group, there was the first control group of stage IIIA(N2) NSCLC patients with NCT (n = 194). This group of patients was given 3 or 4 courses of CT using a regimen similar to the main group: cisplatin / carboplatin + docetaxel / paclitaxel, and in the second control group – similar to the number of courses and drugs, neoadjuvant CT and RT in SOD 50 Gy in adjuvant regimen (n = 60) with further surgical treatments. All patients, depending on the prevalence of the tumor process, underwent radical surgery in the form of lobectomy, bilobectomy or pneumonectomy with mediastinal lymphodissection. Results: median survival in the main group was 30.12 months, in the first control group (NCT) – 23.68 months and the second control group (NCT + adjuvant RT) – 23.23 months, respectively. In the main group, 5 (14.7%) patients were diagnosed with a complete morphological response of the primary tumor and a complete morphological response of the tumor tissue in the regional lymph nodes. In addition to the complete morphological response of the primary tumor and metastatic mediastinal lymph nodes (tumor cells were not detected in histological preparations), in 5 (14.7%) cases, a pronounced morphological response of a highly differentiated tumor was detected (residual viability of tumor tissue to 12±5%). These patients had a higher median survival (45 and 39 months, respectively) compared to controls. Conclusion: the treatment regimen with NCRT is more promising for increasing patient survival.

A Novel Multi-modal Population-graph based Framework for Patients of Esophageal Squamous Cell Cancer Prognostic Risk Prediction.

Prognostic risk prediction is pivotal for clinicians to appraise the patient's esophageal squamous cell cancer (ESCC) progression status precisely and tailor individualized therapy treatment plans. Currently, CT-based multi-modal prognostic risk prediction methods have gradually attracted the attention of researchers for their universality, which is also able to be applied in scenarios of preoperative prognostic risk assessment in the early stages of cancer. However, much of the current work focuses only on CT images of the primary tumor, ignoring the important role that CT images of lymph nodes play in prognostic risk prediction. Additionally, it is important to consider and explore the inter-patient feature similarity in prognosis when developing models. To solve these problems, we proposed a novel multi-modal population-graph based framework leveraging CT images including primary tumor and lymph nodes combined with clinical, hematology, and radiomics data for ESCC prognostic risk prediction. A patient population graph was constructed to excavate the homogeneity and heterogeneity of inter-patient feature embedding. Moreover, a novel node-level multi-task joint loss was proposed for graph model optimization through a supervised-based task and an unsupervised-based task. Sufficient experimental results show that our model achieved state-of-the-art performance compared with other baseline models as well as the gold standard on discriminative ability, risk stratification, and clinical utility. The core code is available at https://github.com/wuchengyu123/MPGSurv.

The Utility of 18F-FDG-PET-CT Metabolic Parameters in Evaluating the Primary Tumor Aggressiveness and Lymph Node Metastasis of Nasopharyngeal Carcinoma.

Following changes in primary tumor (T) and lymph node (N) staging for nasopharyngeal carcinoma (NPC) in the Eighth Edition AJCC Cancer Staging Manual, simplification of T staging has been proposed. However, a limited range of 2-deoxy-2-[fluorine-18] fluoro-D-glucose positron emission tomography-computed tomography (18F-FDG PET-CT) metabolic parameters has been investigated. Therefore, we aimed to evaluate the primary tumor invasiveness and the lymph node metastasis (LNM) of NPC from a metabolic perspective. A total of 435 NPC patients underwent 18F-FDG PET/CT before treatment were retrospectively examined. The primary endpoint was differences in standard uptake value (SUV), lean body mass-normalized SUV (SUL), body surface area-normalized SUV (SUS), glucose-normalized SUV (GN), metabolic tumor volume (MTV), total lesion glycolysis (TLG), and glucose-normalized total lesion glycolysis (GNTLG) of primary tumors and LNM between different T and N stages. The metabolic parameters associated with T and N staging were identified. There were significant differences between all parameters relative to the primary tumor but no significant differences in any parameter relative to the LNM and T stages. Higher mean values of TGNmax, TGNmean, TSUVpeak, and TSUSmax were associated with advanced T stages. Higher mean values of all the LNM parameters were associated with more advanced N stages. Only primary tumor metabolic tumor volume (TMTV), TSUVpeak, TSULmax, and TSUSmax showed a significant positive association with T staging, while lymph node metabolic tumor volume (LNMTV) and TSUSmax were significantly positive in N staging. Our findings suggest that metabolic parameters are useful indicators of tumor invasiveness and LNM based on the Eighth Edition manual. Compared with volume-dependent parameters, TGNmax, TGNmean, TSUVpeak, and TSUSmax may be better indicators of local tumor aggressiveness. SUSmax of the primary tumor was associated with LNM. In addition to SUVmax, other metabolic parameters (eg, SULmax, SUSmax, GNmax, and GNmean) could evaluate tumor aggressiveness and LNM better.

Role of FNAC in Diagnosis of Malignant Lymphadenopathy: A Five-Year Cross-sectional Study at A Tertiary Care Centre, Mumbai, India

Introduction: Fine Needle Aspiration Cytology (FNAC) of the lymph node is a simple tool in the diagnosis of suspected and unsuspected primary and secondary lymph node malignancy. It is a useful first-line investigation to distinguish between infective and malignant lymphadenopathy, especially in a low-resource setting like our country. Aim: To study the utility of FNAC in the diagnosis of lymph node malignancy and to evaluate the efficacy of cytology in diagnosing the primary site of malignancy in case of metastasis. Materials and Methods: A cross-sectional retrospective and prospective study was conducted for a five-year period from July 2015 to June 2020, in the pathology department of Dr. RN Cooper Municipal General Hospital, Mumbai, Maharashtra, India. A total of 112 patients diagnosed with malignancy on FNAC of lymph nodes were studied. The FNAC procedure was performed by cytopathologists using a 23/24G needle attached to a 10 ml syringe. The alcohol-fixed smears were stained with Haematoxylin & Eosin (H&E) stain and Papanicolaou (PAP) stains, while the airdried smears were stained with Giemsa stain. Diagnosis was based on cytomorphological features and clinical presentation. Results: A total of 112 patients were diagnosed with malignancy on FNAC, of which 90 were males and 22 were females, with a male-to-female ratio of 4:1. Cytological analysis of these nodes revealed metastasis in 98 cases (87.50%) and lymphoma in 14 cases (12.50%). The maximum number of patients were in the 51-60 years age group, with 38 cases (33.92%). The most common site of involvement was cervical lymph nodes, with 75 cases (66.96%). The most common cytological diagnosis was metastatic squamous cell carcinoma in 65 cases (58.02%), followed by metastatic un-differentiated carcinoma in 26 cases (23.20%). Conclusion: Fine Needle Aspiration (FNA) is a simple, safe, accurate, cost-effective, and valuable tool in the evaluation of malignant lymphadenopathy. Malignant diseases were easily diagnosed by this simple diagnostic procedure. In the case of a diagnosis of lymphoma, surgery can be completely avoided. It helps in planning further surgical management for metastatic disease, where definitive operative intervention can be performed in one session. In the case of an un-detected primary tumour, FNAC directs further investigations towards the possible primary site.

GPNMB Expression Associates with Inferior Prognosis in Patients with Small Cell Lung Cancer.

Purpose: Small cell lung cancer (SCLC) is widely recognized for its propensity for early and frequent metastases, which contribute to its status as a refractory malignancy. While the high expression of GPNMB in SCLC is well-documented, the precise correlation between GPNMB expression and the prognosis of SCLC remains undetermined. Methods: HTG Edge-seq was used to screen the differential gene expression between primary SCLC lesions and paired metastatic lymph nodes (LN). The plasma concentration of GPNMB was measured using enzyme-linked immunosorbent assay (ELISA). The relationship between GPNMB concentration and clinical characteristics, as well as overall survival (OS) was assessed. One-to-one propensity score matching (PSM) was performed to reduce bias from confounding factors between groups. The invasive, migratory, proliferative, and apoptotic abilities of SCLC cells were evaluated using migration and matrigel invasion assays, CCK8 assay and flow cytometry respectively. Results: GPNMB exhibited a significant up-regulation in LN compared to primary SCLC lesions as determined by HTG Edge-seq. Furthermore, patients with extensive disease demonstrated a significantly elevated plasma GPNMB concentration compared to those with local disease (P = 0.043). Additionally, patients with a high baseline plasma GPNMB level exhibited a shorter OS (10.32 vs. 16.10 months, P = 0.0299). Following PSM analysis, the statistical significance of the difference between the two groups persisted (9.43 vs. 15.27 months, P = 0.0146). Notably, both univariate and multivariate analyses confirmed that higher expression of GPNMB served as an independent biomarker for OS before PSM (P = 0.033, HR = 2.304) and after PSM (P = 0.003, HR = 6.190). Additionally, our study revealed that the inhibition of GPNMB expression through the use of siRNA effectively diminished the metastatic and proliferative capabilities of SCLC. Furthermore, this inhibition resulted in an enhanced ability to induce apoptosis. Conclusions: In light of our findings, it can be inferred that the expression of GPNMB is linked to metastasis and an unfavorable prognosis, thus suggesting its potential as a novel therapeutic target in the treatment of SCLC.

Grade 3 Pancreatic Neuroendocrine Tumor Refractory to Chemotherapy Successfully Treated with Peptide Receptor Radionuclide Therapy Leading to Conversion Surgery.

A 59-year-old woman was diagnosed with a pancreatic neuroendocrine tumor (P-NET; Grade 3, Ki67: 25%) with multiple liver and lymph node metastases and started chemotherapy with streptozosin (500 mg/m2/day) in combination with lanreotide acetate (120 mg). After six courses of (daily) streptozosin, the patient had progressive disease, as assessed by computed tomography (CT), and peptide receptor radionuclide therapy (PRRT) was started as second-line treatment. As PRRT was remarkably successful and the tumor shrank, surgery was performed to resect the primary pancreatic tumor, liver metastases, and lymph node metastases. CT evaluation performed six months after the surgery showed a complete response.

Breast cancer in 80+ year olds.

The risk of breast cancer increases with increasing age. The aim of our retrospective study was to determine the extent of breast and axillary surgery, including subsequent adjuvant therapy, in 80-year and older patients. Between 2017 and 2021, 834 breast cancer patients were operated in the Surgical Department of the EUC Clinic. Ninety-eight women (2× with bilateral cancer) and 2 men were included in this retrospective study. A total of 102 breast cancer cases in patients older than 80 years were analyzed. The surgical procedure corresponded to the stage of the disease and the general condition of the patient. Adjuvant systemic therapy was indicated according to the same principles. At the time of surgery, the patients were more than 80 years old (80-96 years). The predominant type of invasive ductal carcinoma was diagnosed 83×, lobular carcinoma 6×, mucinous 6×, papillary carcinoma 4×, other 3×, with luminal A, B predominating (89×). The breast-conserving procedures were performed 63×. Sentinel node biopsy was performed 65×, supplemented by axillary lymph node dissection 13×. Primary axillary lymph node dissection was performed 15×. No axillary procedure was performed 23×. Radiotherapy was given 49×, chemotherapy 9× and hormonal therapy 82×. Local and regional recurrences were each observed 2×. A total of 37 patients died, 10 of them from breast cancer. The most common cause of death in patients aged 80+ years is a cardiovascular disease, not breast cancer itself. This fact should be taken into account when determining the treatment plan.

Intraoperative Frozen Section Analysis and an Audit of its Diagnostic Accuracy: A Cross-sectional Study from Maharashtra, India

Introduction: Frozen Section (FS) is a diagnostic technique performed intraoperatively to obtain relevant information about the primary diagnosis, margin status, or sentinel lymph nodes, which helps guide the course of surgery. With the prevalent use of FS, it is imperative to recognise and scrutinise its diagnostic pitfalls and make efforts for rectification. Aim: To compare diagnostic results obtained on FS with final paraffin-embedded histopathology and calculate the Diagnostic Accuracy (DA) of FS. Materials and Methods: This cross-sectional study was conducted in the Department of Pathology at a tertiary care hospital, DVVPFs Medical College in Ahmednagar district, Maharashtra, India over a period of two years from January 1, 2020, to December 31, 2021. The diagnosis provided on specimens received for intraoperative FS was compared with the final histopathological diagnosis, considered as the gold standard for the same specimen. The results were categorised as concordant, discordant, and deferred cases. The diagnosis on FS and the final histopathological diagnosis, along with relevant clinical data, were entered into an excel sheet. Further, the DA of FS was calculated. All the discordant cases were analysed. Statistical analysis utilised simple percentage calculations from the excel sheet. Results: A total of 130 cases were analysed using FS, with the most common indication being sentinel lymph node analysis to detect metastatic deposits in 72 (55.4%) cases. Out of the total 130 cases, 119 (91.5%) were concordant, 10 (7.7%) were discordant, and 1 (0.8%) was deferred due to an insufficient specimen. Therefore, the overall DA rate was found to be 91.5% with an error rate of 8.5%. Upon analysis of the 10 discordant cases, the cause of inaccuracy was technical error in 7 (70%) cases and interpretation error in 3 (30%) cases. Conclusion: The DA obtained in the present study was somewhat lower than expected due to technical errors during FS sectioning, leading to artifacts, especially during lymph node processing. Analysis of the discordant cases unveiled this deficit. Therefore, such assessment studies should be performed periodically as they assist in highlighting the shortfalls and provide a plan to boost DA.

Clinical and molecular characteristics of RNF43 mutations as promising prognostic biomarkers in colorectal cancer.

Transmembrane E3 ubiquitin ligase (RNF43) mutations are present in approximately 6-18% of colorectal cancers (CRC) and could enhance Wnt/β-catenin signaling, which is emerging as a promising therapeutic target. This study aims to investigate the clinical and molecular characteristics and potential heterogeneity of RNF43-mutant CRC. A total of 78 patients with RNF43-mutant CRC were enrolled from July 2013 to November 2022. Demographic data, clinical characteristics, treatment regimens used, and survival outcomes were collected and analyzed. Our study uncovered that patients with RNF43 mutations in the N-terminal domain (NTD; n = 50) exhibited shorter overall survival (OS; median months, 50.80 versus not reached; p = 0.043) compared to those in the C-terminal domain (CTD; n = 17). Most RNF43 mutations in NTD had positive primary lymph node status, low tumor mutation burden (TMB-L), and correlated with proficient mismatch repair (pMMR)/microsatellite stable (MSS) status. By contrast, RNF43 mutations in CTD were significantly enriched in deficient MMR (dMMR)/microsatellite instability (MSI-H) tumors with high TMB (TMB-H). N-terminal RNF43-mutated tumors harbored a hotspot variant (RNF43 R117fs), which independently predicted a significantly worse outcome in pMMR/MSS CRC with a median OS of 18.9 months. Patients with RNF43 mutations and the BRAF V600E alterations demonstrated sensitivity to BRAF/EGFR inhibitors. Moreover, we observed that pMMR/MSS patients with RNF43 R117fs mutation had a higher incidence of stage IV, ⩾2 metastatic sites, low TMB, and none of them received PD-1/PD-L1 inhibitor therapy. Our findings provide the first evidence that RNF43 mutations in NTD and the R117fs variant correlate with a poorer prognosis in CRC patients, providing strategies for Wnt-targeted therapy to improve clinical efficacy.

The Role of Fine-needle Aspiration Cytology in Guiding Clinicians for Lymphadenopathy

Abstract Background: Lymphadenopathy is a very common, routinely observed clinical finding which may be caused by a variety of pathological conditions, either a primary lymph node pathology or an underlying systemic disease. Fine-needle aspiration is a simple cost-effective diagnostic tool to screen these lesions, to enhance our knowledge, to understand lymph node pathology, as well as to assist clinicians for an effective management. Aim: The aim of our study was to evaluate the patterns of pathological lesions on lymph node cytology and also to assess any particular trend of diseases in comparison with demographic variables, lymph node size, and consistency in this geographic area. Materials and Methods: In this retrospective study, we studied the reports of 131 lymph node cytology samples collected over a duration of 1 year. All parameters were entered in the Excel sheet and were analyzed for descriptive statistics. Results: The mean age of the patients was 32 years. Reactive lymphadenitis was the most common (42%) pathology, followed by tubercular lymphadenitis, metastatic lymph nodes, and lymphomas. Infectious or inflammatory lesions were noted in younger age groups, whereas malignancies were prominent in elderly patients. Conclusion: This study threw light on diseases, whether infectious, inflammatory, or lymphoproliferative in etiology, which presented with lymphadenopathy in this coastal area and also unveiled the patterns of metastatic lymph nodes, their primary site, and predilection for a particular nodal site.

CCL21 and CLDN11 Are Key Driving Factors of Lymph Node Metastasis in Gastric Cancer.

Gastric cancer (GC) is a leading cause of cancer-related deaths worldwide. Understanding the molecular mechanisms of GC metastasis is crucial for improving patient survival outcomes. RNA sequencing and analysis were performed on tissue samples from primary and lymph node metastatic lesions of gastric cancer. Differential gene analysis and functional pathway analysis were conducted. Immune infiltrating environment and protein expression levels were evaluated using immunohistochemistry. Cell experiments were conducted to investigate the role of CCL21 in GC metastasis. ACTG2, CNN1, DES, MUC6, and PGC were significantly upregulated in primary tumor cells, while CCL21, MS4A1, CR2, CLDN11, and FDCSP were significantly upregulated in metastatic tumor cells. Functional pathway analysis revealed enrichment in pathways related to immune response. CLDN11 and CCL21 were found to play important roles in promoting gastric cancer metastasis. Cell experiments confirmed the role of CCL21 in promoting GC cell growth and metastasis. CCL21 is highly expressed in GC tissues and binds to CCR7, leading to upregulation of CLDN11. This results in GC-lymph node metastasis and abnormal activation of immune cells (B cells and CD4+ T cells). Inhibition of CCL21 and CLDN11 proteins may be a promising strategy for treating GC and preventing lymph node metastasis. These findings provide specific molecular markers for early lymph node metastases of GC, which can aid in developing treatment strategies and predicting patient prognosis.

Pulmonary interstitial lymphography: A prospective trial with potential impact on stereotactic ablative radiotherapy planning for early-stage lung cancer

This prospective feasibility trial investigated pulmonary interstitial lymphography to identify thoracic primary nodal drainage (PND). A post-hoc analysis of nodal recurrences was compared with PND for patients with early-stage lung cancer; larger studies are needed to establish correlation. Exploratory PND-inclusive stereotactic ablative radiotherapy plans were assessed for dosimetric feasibility.

Evaluating Mediastinal Lymph Node Metastasis of Non-Small Cell Lung Cancer Using Mono-exponential, Bi-exponential, and Stretched-exponential Models of Diffusion-weighted Imaging.

To explore and compare the diagnostic values of mono-exponential, bi-exponential, and stretched-exponential diffusion-weighted imaging (DWI) parameters of primary lesions and lymph nodes (LNs) to predict mediastinal LN metastasis in patients with non-small cell lung cancer. Sixty-one patients with non-small cell lung cancer underwent preoperative magnetic resonance imaging, including multiple b -value DWI. The DWI parameters, including apparent diffusion coefficient (ADC) from a mono-exponential model, true diffusion (D) coefficient, pseudo-diffusion (D*) coefficient, and perfusion fraction (f) from a bi-exponential model, distributed diffusion coefficient (DDC) and intravoxel diffusion heterogeneity index (α) from a stretched-exponential model of primary tumors and LNs and the size characteristics of LNs, were measured and compared. Multivariate logistic regression analysis was used to establish models for predicting mediastinal LN metastasis. Receiver operating characteristic analysis was applied to evaluate diagnostic performances. The DWI parameters of primary tumors showed no statistical significance between LN metastasis-positive and LN metastasis-negative groups. Nonmetastatic LNs had significantly higher ADC, D, DDC, and α values compared with metastatic LNs (all P < 0.05). The short-dimension, long-dimension, and short-long dimension ratio of metastatic LNs was significantly larger than those of nonmetastatic ones (all P < 0.05). The D value showed the best diagnostic performance among all DWI-derived single parameters, and the short dimension of LNs performed the same among all the size variables. Furthermore, the combination of DWI parameters (ADC and D) and the short dimension of LNs can significantly improve diagnostic efficiency. The ADC, D, DDC, and α from the mono-exponential, bi-exponential, and stretched-exponential models were demonstrated efficient in differentiating benign from metastatic LNs, and the combination of ADC, D, and short dimension of LNs may have a better diagnostic performance than DWI or size-derived parameters either in combination or individually.

Locoregional Ablative Radiation Therapy for Patients With Breast Cancer Unsuitable for Surgical Resection

Breast cancer patients unsuitable for surgical resection are typically managed with palliative systemic therapy alone. We report outcomes of 5-fraction ablative radiotherapy for non-resected breast cancers. Retrospective analysis of an institutional registry of breast cancer patients unsuitable for resection who underwent 35-40 Gy/5 fractions to the primary breast tumour and/or regional lymph nodes from 2014-2021. Primary outcomes were cumulative incidence of local failure (LF) and grade ≥3 toxicity (CTCAE v5.0). We reviewed 57 patients who received 61 treatment courses (median age of 81 years, range 38-99). Unresectable tumour (10%), patient refusal (18%), medical inoperability (35%), and metastatic disease (37%) were the causes of not having surgery. Five patients (8%) had previously undergone adjuvant locoregional radiotherapy. Fifty-four percent (n=33/61) of treatment courses targeted the breast only, 31% (n=19/61) both the breast and lymph nodes, and 15% (n=9/61) the lymph nodes only. Sixty-seven percent (n=35/52) of the courses that targeted the breast were delivered with partial breast irradiation and 33% (n=17/52) with whole breast radiotherapy (median dose of 25Gy in 5 fractions) ± simultaneous integrated boost to the primary tumour. Most primary tumours (65%, n=34/52) and target lymph nodes (61%, n=17/28) were treated with a dose of 35 Gy in 5 fractions. Most treatments (52%) were delivered with intensity-modulated radiotherapy (IMRT). Radiotherapy was delivered daily (20%), every other day (18%), twice weekly (36%) or weekly (26%). The 2-year cumulative incidence of LF was 11.4% and grade≥3 toxicity was 15.1%. The grade ≥3 toxicity was 6.5% for IMRT treatments, versus 7.7% for non-IMRT treatments targeting partial breast and/or lymph nodes (HR 1.13, P=0.92), versus 38.9% for non-IMRT treatments targeting the entire breast (HR 6.91, P=0.023). All grade ≥3 toxicity cases were radiation dermatitis. No cases of brachial plexopathy were observed. 35-40 Gy in 5 fractions is a safe and effective breast stereotactic body radiotherapy regimen and may be an attractive option for patients who are not surgical candidates. Highly conformal techniques (i.e. IMRT or partial breast irradiation) were associated with a reduced risk of toxicity and should be the preferred treatment approaches.

Optimization of algorithms for in vivo preclinical screening of compounds with an alleged antitumor effect

BACKGROUND: Despite a large number of publications on preclinical studies of compounds with a suspected antitumor effect on in silico and in vitro models, in vivo studies are the most informative. The experimental part of the work on laboratory animals has its own peculiarities in the field of preclinical research: a large number of animals and analog compounds in the series, two- and three-phase staging, and, consequently, high cost and labor-intensive execution.&#x0D; AIM: Optimization of algorithms for screening preclinical in vivo studies of compounds with a suspected antitumor effect.&#x0D; MATERIALS AND METHODS: To form the algorithm in the preclinical study, primary data was obtained using standard pharmacological (determination of the toxicity class index, antitumor and antimetastatic activity, inhibition of tumor growth by weight, average life expectancy of animals) and morphological (autopsy, micropreparations with hematoxylin and eosin staining, as well as immunohistochemical study using monoclonal antibodies) methods with the selection of leading compounds for in-depth study with a description of the mechanism of pharmacological activity.&#x0D; RESULTS: Based on a series of comparative experiments, the following algorithm of preclinical in vivo study for newly synthesized compounds with an alleged antitumor effect has been tested.&#x0D; Stage 1. Determination of the toxicity class with a single intragastric administration to Wistar rats according to the Organization for Economic Co-operation and Development 420 protocol and selection of candidates for antitumor drugs according to the principle of the greatest safety of use.&#x0D; Stage 2. Determination of the presence/absence of pharmacological activity of the tested compounds. Compounds of toxicity classes IV and V (according to the Globally Harmonized System of Hazard Classification and Labeling of Chemical Products) in a wide range of doses (doses are selected depending on the toxicity class) are examined for pharmacological activity before the natural death of tumor-bearing animals with the identification of leader substances, in-depth study of which is appropriate. The selection of promising substances and total doses for administration at the next stage is determined by the life expectancy of tumor-bearing animals.&#x0D; Stage 3. Determination of indicators of antitumor and antimetastatic activity of leader substances with a fixed euthanasia period for all tumor-bearing animals and determination of possible mechanisms for the implementation of the therapeutic effect using immunohistochemical analysis.&#x0D; Stage 4. To study the effect of the tested compounds on the growth rates of the primary tumor node and metastatic foci at different stages of the development of the tumor process, when administered in different modes, as part of combined and monochemotherapy, with mandatory clarification of the mechanisms for the implementation of antitumor and antimetastatic activity using biochemical and immunohistochemical techniques.&#x0D; Stage 5. The study of the most promising compounds according to the guidelines for preclinical safety studies for the purpose of clinical trials and registration of medicines (the document was approved by the decision of the Board of the Eurasian Economic Commission of November 26, 2019, N 202): «Toxicity studies with repeated (multiple) administration of the drug, preclinical studies conducted in order to justify the conduct of exploratory clinical studies, studies of local tolerability of the drug, studies of genotoxicity of the drug, carcinogenicity of the drug, etc.»&#x0D; CONCLUSIONS: The step-by-step exclusion of the tested compounds from the line of similarly-structured substances described by us will increase the efficiency of selecting promising candidates for antitumor drugs and reduce the cost of conducting preclinical studies of compounds with an alleged antitumor effect.

A robust hybrid model with low energy consumption for biosensor nano-networks

The biosensor nano-network consists of mobile nano-nodes, nano-routers and micro-gateways in various levels of hierarchy. In the human body, the energy is harvested by the nano-devices by transforming various forms of energy into electrical energy. Simultaneously, the energy is consumed to collect and transmit data to the external network. So, a proper balance between energy harvesting and energy consumption is required. The current multi-path routing enables data flooding, using energy from numerous redundant nodes along the path. Whereas, single-path routing has the drawback of packet loss. In this article, we propose a hybrid model with low energy consumption and minimal packet loss. We grouped the nano-nodes into clusters by which the redundancy is avoided to a great extent. The nano-nodes communicate with a cluster primary node using a Lightweight Secure Directed Diffusion multi-path protocol, and the primary nodes communicate with the nano-router using Energy Conserving Routing single-path protocol at the layer-1 of our model. In the conventional models, the micro-gateway communication with the nano-routers created energy irregularities. So, we come up with a query routing protocol at the layer-2 that chooses a particular nano-router for communication with the micro-gateway. As a result, the energy consumption at the micro-gateway is decreased.