Background

Cancer cells express immunosuppressive molecules, such as programmed death ligands (PD-L)1 and PD-L2, enabling evasion from the host’s immune system. Cancer cells synthesize and secrete acetylcholine (ACh), acting as an autocrine or paracrine hormone to promote their proliferation, differentiation, and migration.

Methods

We correlated the expression of PD-L1, PD-L2, cholinergic muscarinic receptor 3 (M3R), alpha 7 nicotinic receptor (α7nAChR), and choline acetyltransferase (ChAT) in colorectal cancer (CRC) tissues with the stage of disease, gender, age, risk, and patient survival. The effects of a muscarinic receptor blocker, atropine, and a selective M3R blocker, 4-DAMP, on the expression of immunosuppressive and cholinergic markers were evaluated in human CRC (LIM-2405 and HT-29) cells.

Results

The increased expression of PD-L1, M3R, and ChAT at stages III-IV was associated with a high risk of CRC and poor survival outcomes independent of patients’ gender and age. α7nAChR and PD-L2 were not changed at any CRC stages. Atropine and 4-DAMP suppressed the proliferation and migration of human CRC cells; induced apoptosis; and decreased PD-L1, PD-L2, and M3R expression in CRC cells via the inhibition of EGFR and the phosphorylation of ERK.

Conclusions

The expression of immunosuppressive and cholinergic markers may increase the risk of recurrence of CRC. These markers might be used in determining prognosis and treatment regimens for CRC patients. Blocking cholinergic signaling may be a potential therapeutic strategy for CRC through anti-proliferation and anti-migration via the inhibition of EGFR and the phosphorylation of ERK. These effects allow the immune system to recognize and eliminate cancer cells.

Introduction: One of the main causes of death worldwide is cancer. The capacity of theranostic prodrugs to provide simultaneous diagnosis and therapy has drawn more and more attention in recent years. Here, we present the development of a novel theranostic molecule, called GF, based on the fluorescent dye 5(6)-carboxyfluorescein and the nucleoside anticancer drug gemcitabine.

Material and Methods: By using confocal microscopy to assess the synthesized GF's mode of cell uptake and cellular localization, a clathrin-mediated endocytosis was also revealed. Furthermore, three distinct human cancer lines—A549 (lung cancer), U87 and T98 (glioblastoma) were used to investigate the in vitro cytotoxicity and its pharmacokinetic profile in mice. Lastly, the effectiveness of the treatment in vivo was assessed using zebrafish and xenografted mice.

Results: GF has the ability to effectively enter tumor cells and locate in the lysosomes, which are the site of drug release and anticancer action. In mice, GF demonstrated improved pharmacokinetics as compared to gemcitabine. Higher concentrations of GF had notable anti-proliferative effects; yet, at all tested concentrations in all cell lines, GF was shown to be less cytotoxic than gemcitabine.

Conclusion: These findings suggest that GF, as a prodrug, can be a highly effective cytotoxic agent against glioblastoma. Additionally, as a potential gemcitabine prodrug, GF demonstrated less cytotoxic action than gemcitabine.

Abstract: Hypoxia is a key component of the tumour microenvironment, and it is linked to cell proliferation, angiogenesis, metabolism, and the tumour immune response. All of these variables may accelerate tumour growth, increase tumour aggressiveness, raise tumour metastatic potential, and result in a poor prognosis. The goal of this trial was to determine if the pharmaceutical substance Ometaxine mepesuccinate, commonly known as Homoharringtonine or (HHT), is effective. Cephalotaxus fortunei is the source of natural plant alkaloids. HHT is a chemical derived from traditional Chinese medicine that has been licenced by the Food and Drug Administration for the treatment of leukaemia. Previous research has suggested that HHT might limit protein synthesis and promote apoptosis by upregulating the proapoptotic protein Bax and triggering the caspase-3-mediated cleavage of PARP. Aside from hematologic tumours, HHT has also been shown to be useful in renal cell carcinoma, colon rectal cancer, and non-small cell lung cancer. The impact of HHT on CRC, as well as the underlying HIF-1/VEGF-associated mechanisms of action, have not been explored. Tube formation was used to measure in vitro angiogenesis. A cell viability assay, scratch assay, real-time PCR, enzyme-linked immunosorbent assay and immunoblotting were used to analyse the gene and protein expressions of hypoxia-inducible factor-1 (HIF-1) and vascular endothelial growth factor (VEGF). The development of hypoxic HUVEC cell tubes is inhibited by HHT. In CRC SW480 cells, HHT reduced hypoxia-induced VEGF overexpression at both the mRNA and protein levels. Furthermore, HHT suppressed HIF-1a-induced angiogenesis in vitro via inhibiting HIF-induced ERK phosphorylation. Hypoxia raised HIF-1 mRNA and protein levels, which were all suppressed by HHT. Hypoxia significantly elevated ERK and AKT phosphorylation in CRC cells, which HHT reduced. By reducing HIF-1/VEGF-mediated intercellular communication between CRC cells and endothelial cells, HHT decreased hypoxic CRC cell-induced angiogenesis. We think that HHT could be an ideal candidate for CRC tumour inhibition and needs more investigation for further study.

Background

Oxidative stress and immunoinflammation are two important areas in the field of cancer research. At present, no studies have confirmed that both can accurately predict the progression and long-term prognosis of gastric cancer (GC).

Methods

In this study, univariate and multivariate Cox proportional hazards regression analyses were used to analyze oxidative stress and immunoinflammatory indicators in the training set, and the best biomarkers for the establishment of an improved systemic prognosis score (mSPS) were screened. The Area Under the Curve (AUC) and Concordance Index (C-index) were used to evaluate the predictive ability of our model for long-term efficacy. Kaplan–Meier (K-M) survival analysis was conducted to assess survival outcomes. Univariate and multivariate Cox proportional hazards regression analyses were performed to investigate the impact of the model on tumor progression, lymph node metastasis, and overall prognosis. These findings were subsequently validated using both internal and external validation sets.

Results

A total of 3788 GC patients were included in this study, including 2339 in the training set, 1003 in the internal validation set, and 446 in the external validation set. The Receiver Operating Characteristic (ROC) curve analysis revealed that the AUC for mSPS in predicting overall survival (OS) among GC patients significantly surpassed that of all other markers related to oxidative stress and immunoinflammatory indicators (P < 0.001). Compared to the low mSPS group, patients in the high mSPS group were predominantly male and were more likely to be aged over 65, exhibit a higher BMI, possess a higher ASA grade, display a greater prevalence of vasculature and neuroinfiltration, exhibit deeper tumor invasion, have a higher incidence of lymph node metastasis, be diagnosed at a more advanced tumor stage (Stage III), and receive a higher proportion of chemotherapy (P < 0.05). Logistic regression analysis demonstrated that elevated mSPS was an independent risk factor for tumor invasion and lymph node metastasis in patients with GC. COX regression analysis further indicated that high mSPS independently contributed to a poorer prognosis among GC patients. The association between mSPS and pathological stage, as well as long-term prognosis, was validated through both internal and external sets, using the same cut-off value. The results aligned consistently with the training set effect.

Conclusions

As a newly developed prognostic score based on oxidative stress and immunoinflammatory indicators, mSPS can independently and effectively predict the tumor burden of advanced gastric cancer, and further distinguish the long-term prognosis of GC patients based on a pTNM staging system.

Background:
Lung carcinoma is the leading cause of cancer-related mortality, with 50% of cases presenting metastases at the time of diagnosis. Metastatic peritoneal localisation of lung carcinoma is an exceptionally rare situation, accounting for 3% of lung cancers. The rarity is heightened when dealing with small cell lung carcinoma cases, making reported instances even more exceptional.
Accurate diagnosis and proper disease staging are crucial in determining whether the disease is metastatic or non-metastatic, influencing subsequent management.

A 58-year-old patient with a history of non-metastatic small cell lung carcinoma diagnosed in October 2021 (2 years ago) underwent radiotherapy and achieved remission. In February 2023, the patient experienced abdominal pain and hematemesis with melena. Esophagogastroduodenoscopy revealed an ulcerative, polypoid gastric process. Biopsy confirmed moderately differentiated gastric adenocarcinoma. A whole-body CT scan identified new peritoneal nodules.
Laparoscopy with biopsy was performed, and specimens were sent for histopathological examination.

Results:
Histologically, the specimen showed a diffuse tumor proliferation consisting of generally small, highly mitotic cells, displaying irregular, hyperchromatic nuclei and scant, basophilic cytoplasm. Immunohistochemical analysis revealed the following:
-Positive staining of tumor cells for TTF1 and CK7 and negative staining for CK20, indicating a pulmonary origin and ruling out gastric origin.
-Tumor cells expressed chromogranin A and synaptophysin, with a high proliferative index (Ki67) estimated at 60%.
Considering all data, the diagnosis retained is a secondary localisation of undifferentiated carcinoma, with immunohistochemical profiling primarily aligning with small cell lung carcinoma.

Discussion and Conclusions:
Small cell carcinoma, a therapeutic emergency, poses a significant health problem due to its high incidence and mortality. Diagnosis relies on histology supported by immunohistochemistry. Patients often present acute symptoms due to the rapid local growth of intrathoracic tumors, especially at the pulmonary hilum, or distant extrapulmonary dissemination in 40% of cases involving lymph nodes, liver, adrenal glands, bones, or brain.
Peritoneal metastasis is exceptionally rare, with the most common histological type being pulmonary adenocarcinoma (80% of cases), and the least common scenario is the one reported in this abstract: small-cell carcinoma.
Thorough morphological and immunohistochemical studies, guided by the suspected histological type and patient history, are mandatory for a conclusive diagnosis.

Introduction
Radiotherapy can improve the survival rates of patients with gliomas; meanwhile, the impaired cognitive functions have been brought to the forefront, with the offending organ, radiosensitive hippocampus. Moreover, there is no further research exploring whether we can minimize the radiation dose to the hippocampus using volumetric-modulated arc therapy (VMAT) as the treatment for gliomas. This study aimed to assess the feasibility of hippocampus-sparing volumetric-modulated arc therapy (HS VMAT) in patients with WHO Grade II gliomas.

Materials & Methods
Hippocampus-sparing VMAT plans and non-hippocampus-sparing VMAT plans were generated using a computed tomography (CT) dataset of 10 patients with WHO grade II gliomas who underwent postoperative radiotherapy. The gross target volume (GTV), organs at risk (OARs), and hippocampus were localized based on the CT images and modified according to the pre-treatment MRI images in fusion. The prescribed dose for both radiotherapy plans was 54Gy/30F to primary tumor volume (PTV). The dose volume histogram (DVH), homogeneity index (HI), conformity index (CI), and irradiated dose of the hippocampus and other OARs were analyzed.

Results
The HI (0.057±0.014 vs 0.046±0.009, p=0.011) and CI (0.91±0.02 vs 0.92±0.02, p=0.071) for both HS VMAT plans and NHS VMAT plans were clinically acceptable. Regarding the protection of OARs, HS VMAT plans were equally capable and even lowered the radiation dosages to the brainstem (35.56±16.67 vs 41.74±16.18, p=0.017) and spinal cord (1.34±0.77 vs 1.43±0.80, p=0.006). The HS VMAT plans substantially reduced radiation doses to the hippocampus. The ipsilateral hippocampal Dmin, Dmean, Dmax, and D40% for NHS VMAT plans and HS VMAT plans were 22.33±19.12 vs. 7.80±4.20 (p=0.005), 35.20±19.58 vs. 18.60±8.52 (p=0.005), 45.37±16.84 vs. 34.28±12.79 (p=0.013), and 36.73±19.52 vs. 19.98±9.63 (p=0.005). While the contralateral hippocampal Dmin, Dmean, Dmax, and D40% of the two plans were 9.68±6.51 vs. 3.38±1.12 (p=0.005), 16.86±8.86 vs. 4.86±0.91 (p=0.006), 29.13±13.88 vs. 8.41±1.38 (p=0.001), and 17.30±9.14 vs. 4.89±0.94 (p=0.005), respectively.

Conclusion
The use of the HS VMAT plan is feasible, which can effectively reduce the dosage delivered to the hippocampus while not significantly exacerbating the HI, CI, and OARs. Consequently, it may help mitigate the risk of cognitive impairment to a certain extent.

[Background] Boron neutron capture therapy (BNCT) is a particle beam therapy that enables the precise targeting of tumors at the cellular level. Building on the success observed in nuclear reactors, BNCT exhibits potential as a therapeutic approach for managing invasive brain tumors, including malignant gliomas and high-grade meningiomas. Metastatic spinal tumors have been treated by surgical removal, radiation therapy, and a variety of other multidisciplinary therapies. However, none of them has been successful in overcoming this metastatic disease. This study aimed to assess the efficacy of BNCT in treating metastatic spinal tumor using a mouse model.

[Methods] In vitro experiment: neutron irradiation was applied to A549 human lung carcinoma cells with (BNCT group) or without p-boronophenylalanine (BPA) 10 µg Boron/ml for 24 h exposure before irradiation (hot control group) for 0 to 30 min. The cell killing effect was assessed by a colony forming assay. In vivo experiment: an A549 subcutaneous tumor was removed and implanted into the lumbar lamina, where it was partially resected. A few weeks later, metastatic spinal tumor models were successfully established. For the biodistribution of boron, after 2.5, 6, or 24 h of intravenous BPA administration (250mg/ mouse body weight), the metastatic spinal tumor models were sacrificed and the boron concentration of the tumor and each organ measured using Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-AES).

[Results] In vitro neutron experiment: the BNCT group showed a greater cell killing effect than the hot control group. The highest boron accumulation in the tumor was observed at 2.5 h after BPA administration (8.6 µg B/g), and the tumor to normal spinal cord ratio (T/N) was 2.7 and the tumor to blood ratio (T/Bl) was 2.8.

[Conclusions] The findings may indicate that BNCT represents an effective approach in the management of metastatic spinal tumor. We will perform an in vivo neutron irradiation experiment on this model to evaluate the effects of BNCT on survival and neurological function.

Exposure to mycovirus-containing Aspergillus flavus alters transcription factors in normal and leukemia cell lines

Abstract: Transcription factors bind specific DNA motifs to regulate the expression of target genes in order to maintain normal hematopoiesis. The dysregulation of these factors can lead to hemopoietic disorders, including leukemias. Mutations, translocations, or the aberrant expression of several transcription factors in various leukemias have been well documented. Mycoviruses are known to alter the genetics of their fungal host and transform their biological characteristics. We have evaluated the effects of the products of a certain mycovirus-containing Aspergillus flavus (MCAF), which we have isolated from the home of a patient with leukemia, on the transcription factors of acute lymphoblastic (ALL), acute myelogenous leukemia (AML) cell lines, and controls. This organism does not produce any aflatoxin. Our previously published studies have shown that patients with B-cell ALL, unlike controls, have antibodies to MCAF, and the exposure of the mononuclear leukocytes from ALL patients in complete remission to its products results in the re-development of genetic and cell-surface markers characteristic of ALL. In additional investigations, ALL and AML cell lines and controls were exposed to incremental doses of the supernatant of the culture of MCAF. Prior to and after exposure, the levels of PAX-5 and Ikaros (75 and 55 kDa) in ALL cell lines, and AML-1,c/EBP-a and PU.1 in AML cell lines, as well as NF-κB(p65) transcription factors in both cell lines, were assessed. Cellular viability was also evaluated. The exposure of the ‘normal’ cell line to the MCAF resulted in apoptosis, and the downregulation of all tested transcription factors, with no detectable levels remaining. In leukemia cell lines, exposure to MCAF also caused cell death; however, while there was a downregulation of all tested transcription factors, some residual levels were retained. The genetic alterations caused by MCAF are novel findings and of significance. Further studies on the possible role of mycovirus-containing Aspergillus flavus in leukemogenesis are warranted.

Background: Surgical quality control is a crucial determinant for evaluating tumor efficacy.

Objective: To assess the ClassIntra grade for quality control and oncological outcomes of robotic radical surgery for gastric cancer (GC).

Methods: Data of patients undergoing robotic radical surgery for GC at a high-volume center were retrospectively analyzed. Patients were categorized into two groups, the iAE (intraoperative adverse event) group and the non-iAE group, based on the occurrence of intraoperative adverse events. The iAEs were further classified into five sublevels (ranging from I to V according to severity) based on the ClassIntra grade. Surgical performance was assessed using the Objective Structured Assessment of Technical Skill (OSATS) and the General Error Reporting Tool.

Results: This study included 366 patients (iAE group: n=72 [19.7%] and non-iAE group: n=294 [80.3%]). The proportion of ClassIntra grade II patients was the highest in the iAE group (54.2%). In total and distal gastrectomies, iAEs occurred most frequently in the suprapancreatic area (50.0% and 54.8%, respectively). In total gastrectomy, grade IV iAEs were most common during lymph node dissection in the splenic hilum area (once for bleeding [grade IV] and once for injury [grade IV]). The overall survival (OS) and disease-free survival (DFS) of the non-iAE group were significantly better than those of the iAE group (Log rank P<0.001). Univariate analysis showed that tumor location, surgical procedure, tumor size, cT, cN, and OSATS scores were significantly associated with an increased risk of iAE (all P<0.05). A further multivariate analysis showed that BMI≥28 (OR=4.70), cT_2-4 (OR=4.15), and cN₊ (OR=3.28) were independent risk factors for an increased incidence of iAE (all P<0.05), whereas a high OSATS score was an independent protective factor for iAE (OR=0.62, P<0.001).

Conclusion: iAEs in patients who underwent robotic radical gastrectomy significantly correlated with the occurrence of postoperative complications and a poor long-term prognosis. Postoperative complications not only affect the postoperative recovery outcome but also the choice of the next treatment plan, due to the presence of serious and prolonged complications. The iAE group had advanced tumor stages, and high-risk factor analysis showed that they were more likely to experience iAEs. Therefore, the utilization and inclusion of ClassIntra grading as a crucial surgical quality control and prognostic indicator in the routine surgical quality evaluation system are recommended.

Purpose: FDG-PET is often used in the assessment, treatment planning, and monitoring of locally advanced cervical cancer (LACC), contributing to improved patient care and management. The aim of this study is to examine the utility of FDG-PET radiomics in discriminating between patients with LACC who demonstrated an early response to treatment and those who did not.

Methods: The study uses FDG-PET scans of 21 patients obtained from The Cancer Imaging Archive (TCIA) Cervical Cancer Tumor Heterogeneity (CCTH) collection. Among the study cohort, 12 patients responded to chemoradiation treatment, while the remaining 9 did not. A total of 59 radiomics texture features were extracted from the gross tumor volumes (GTVs). Top radiomics features in terms of differentiating response status were selected using a sequential feature selection (SFS) algorithm. The top two features were used to build a support vector machine (SVM) predictive classifier, with performance assessed by a receiver operating characteristic (ROC) curve analysis.

Results: The top two radiomics features being selected consisted of the gray level co-occurrence matrix (GLCOM)-based cluster prominence and gray level run length matrix (GLRLM)-based length nonuniformity. The SVM predictive classifier built using the top two features had an area under the ROC curve (AUC) of 0.88, showing strong discriminative power between responders and non-responders.

Conclusions: The radiomic features extracted from FDG-PET scans showed a discriminative ability to discern patients with LACC who would have an early response to treatment. This knowledge could have clinical implications for adjusting treatment strategies for patients who might not show an early response.