Distinct immunoreactions after a primary tumor microwave ablation using different heating parameters in a VX2 tumor model.

BACKGROUND: Thermal ablation of solid tumors in situ can activate the immune system and produce a specific immune response against the tumor. Microwave ablation (MWA) with different parameters can ablate tumors with similar sizes and cause different local inflammatory effects. Our aim was to determine the immunological effects induced by different energy modes of MWA for a primary tumor. METHODS: Seventy rabbits with VX2 tumors that were implanted subcutaneously underneath the right second nipple were treated with high-power MWA (40 W for 1 min), low-power MWA (20 W for 2 min), or surgical resection or were left without treatment (control). Survival time was evaluated by log-rank test. On day 14 after ablation, immunohistochemistry and flow cytometry were used to evaluate the T-cell immune responses. In addition, the cytokine patterns were identified by enzyme-linked immunosorbent assay. RESULTS: Tumor eradication was achieved completely in the MWA groups, as proven by nicotinamide adenine dinucleotide diaphorase staining. Compared with the three treatment groups, the control group had a significantly higher number of pulmonary metastases and worse survival; however, no significant difference was observed among the three treatment groups. More intra-tumoral and systemic CD4+ and CD8+ T-cells were induced in the MWA groups than in the control group. Compared with operation, MWA induced more systemic CD4+ T-cells. More intra-tumoral CD4+ and CD8+ T-cells and systemic CD4+ T-cells were induced by high-power MWA than by low-power MWA. Moreover, MWA increased the interleukin 2 (IL2) and IL12 levels and decreased the IL4, IL6, and IL10 levels. Importantly, the serum IL12 level was significantly higher after high-power MWA than after low-power MWA. CONCLUSION: High-power MWA enhanced the type 1 T helper immune response and may be selected for the treatment of solid tumors. Future studies are needed to confirm our results.

Role of GSDM family members in airway epithelial cells of lung diseases: a systematic and comprehensive transcriptomic analysis.

Gasdermin (GSDM) family, the key executioners of pyroptosis, play crucial roles in anti-pathogen and anti-tumor immunities, although little is known about the expression of GSDM in lung diseases at single-cell resolution, especially in lung epithelial cells. We comprehensively investigated the transcriptomic profiles of GSDM members in various lung tissues from healthy subjects or patients with different lung diseases at single cell level, e.g., chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), lung adenocarcinoma (LUAD), or systemic sclerosis (SSC). The expression of GSDM members varied among pulmonary cell types (immune cells, structural cells, and especially epithelial cells) and even across lung diseases. Regarding disease-associated specificities, we found that GSDMC or GSDMD altered significantly in ciliated epithelia of COPD or LUAD, GSDMD in mucous, club, and basal cells of LUAD and GSDMC in mucous epithelia of para-tumor tissue, as compared with the corresponding epithelia of other diseases. The phenomic specificity of GSDM in lung cancer subtypes was noticed by comparing with 15 non-pulmonary cancers and para-cancer samples. GSDM family gene expression changes were also observed in different lung epithelial cell lines (e.g., HBE, A549, H1299, SPC-1, or H460) in responses to external challenges, including lipopolysaccharide (LPS), lysophosphatidylcholine (lysoPC), cigarette smoking extract (CSE), cholesterol, and AR2 inhibitor at various doses or durations. GSDMA is rarely expressed in those cell lines, while GSDMB and GSDMC are significantly upregulated in human lung epithelia. Our data indicated that the heterogeneity of GSDM member expression exists at different cells, pathologic conditions, challenges, probably dependent upon cell biological phenomes, functions, and behaviors, upon cellular responses to external changes, and the nature and severity of lung disease. Thus, the deep exploration of GSDM phenomes may provide new insights into understanding the single-cell roles in the tissue, regulatory roles of the GSDM family in the pathogenesis, and potential values of biomarker identification and development.

ACAT1-mediated METTL3 acetylation inhibits cell migration and invasion in triple negative breast cancer.

Triple-negative breast cancer (TNBC) is a heterogeneous and aggressive disease with poor prognosis. Acetylation modifications affect a great number of biological processes of malignant tumors. The current study aims at revealing the role of acetylation-related mechanism in TNBC progression. Methyltransferase like-3 (METTL3) was found to be downregulated in TNBC cells via quantitative polymerase chain reaction (qPCR) and western blot analyses. Co-Immunoprecipitation (Co-IP) and GST pulldown assays revealed the interaction between acetyl-CoA acetyltransferase 1 (ACAT1) and METTL3. Through further immunoprecipitation (IP) assay, we determined that ACAT1 stabilizes METTL3 protein via inhibiting the degradation of ubiquitin-proteasome. Functionally, ACAT1 inhibits TNBC cell migration and invasion. Moreover, nuclear receptor subfamily 2 group F member 6 (NR2F6) regulates ACAT1 expression at transcriptional level. Finally, we demonstrated that NR2F6/ACAT/METTL3 axis suppresses the migration and invasion of TNBC cells via METTL3. In conclusion, NR2F6 transcriptionally activates ACAT1 and promotes the suppressive effects of ACAT1-mediated METTL3 acetylation on TNBC cell migration and invasion.

Risk factors of recurrent thyroid nodules after radiofrequency ablation.

Objective: To investigate the risk factors of thyroid nodule recurrence after radiofrequency ablation (RFA). METHODS The medical record information of 120 patients with thyroid nodules admitted to our hospital from June 2019 to April 2022 was retrospectively analysed. All participants received RFA treatment. According to the results of the postoperative thyroid ultrasound examination (USG), the patients were divided into the recurrence group (R, N=16) and the non-recurrence group (NR, N=104). Binary logistic regression analysis was performed to identify the independent risk factors of thyroid nodule recurrence after RFA. The receiver operating characteristic (ROC) curve was used to analyse the value of the forecast of each independent factor and combined model for thyroid nodule recurrence after RFA. Results: During the follow-up period, 16 patients recurred, and the recurrence rate was 13.33%. Univariate regression analysis showed that whether the nodules are solitary (WNS), nodule diameter (ND), the degree of risk of nodular location (DRN), recurrent laryngeal nerve (RLN) injury were associated with thyroid nodule recurrence after RFA (P<0.05). Binary logistic regression analysis showed that WNS, ND, DRN and RLN injury were independent risk factors for the recurrence of thyroid nodules after RFA (P<0.05). ROC analysis of independent factors and combined model showed that solitary nodules, nodule diameter and nodule location risk degree had diagnostic value, while RLN injury had no predictive value. The combined model is more predictive than the independent factors. Conclusions: The risk factors of recurrent thyroid nodules after radiofrequency ablation are related to WNS, ND, DRN and so on, which should be paid attention to and preventive measures should be taken.

Blood metagenomics next-generation sequencing has advantages in detecting difficult-to-cultivate pathogens, and mixed infections: results from a real-world cohort.

Background: Blood is a common sample source for metagenomics next-generation sequencing (mNGS) in clinical practice. In this study, we aimed to detect the diagnostic value of blood mNGS in a large real-world cohorts. Methods: Blood mNGS results of 1,046 cases were collected and analyzed along with other laboratory tests. The capabilities and accuracy of blood mNGS were compared with other conventional approaches. Results: Both the surgical department and the intensive care unit had a positive rate of over 80% in blood mNGS. The positive rate of mNGS was consistent with clinical manifestations. Among the 739 positive samples, 532 were detected as mixed infections. Compared to pathogen cultures, the negative predictive value of blood mNGS for bacteria and fungi detection was 98.9% [95%CI, 96.9%-100%], with an accuracy rate of 89.39%. When compared with polymer chain reaction, the consistency rates of blood mNGS for virus identification were remarkably high. Conclusions: Blood mNGS have significant advantages in detecting difficult-to-cultivate bacteria or fungi, viruses, and mixed infections, which benefits patients of surgery department the most. Samples other than blood are recommended for mNGS test if a specific infection is suspected. The reporting threshold and reporting criteria of blood mNGS need to be optimized.

Research progress on microRNA-1258 in the development of human cancer.

microRNAs (miRNAs) are small endogenous RNAs composed of 20-22 nucleotides that do not encode proteins, which regulate the expression of downstream genes by targeting the 3' untranslated region of mRNA. Plentiful research has demonstrated that miRNAs participate in the initiation and development of diverse diseases and malignant tumors. miR-1258 exerts great influence on tumors, including tumor growth, distant metastasis, migration, invasion, chemosensitivity, cell glycolysis, apoptosis, and stemness. Interestingly, miR-1258 is a miRNA with explicit functions and has been investigated to act as a tumor suppressor in studies on various types of tumors. With accumulating research on miR-1258, it has been found to be used as a biomarker in the early diagnosis and prognosis prediction of tumor patients. In this review, we outline the development of miR-1258 research, describe its regulatory network, and discuss its roles in cancer. Additionally, we generalize the potential clinical applications of miR-1258. This review offers emerging perspectives and orientations for further comprehending the function of miR-1258 as a diagnostic and prognostic biomarker and potent therapeutic target in cancer.

Analysis of Negative Results of Metagenomics Next-Generation Sequencing in Clinical Practice.

Background: Metagenomics next-generation sequencing (mNGS) has been increasingly used in the clinic, which provides a powerful tool for the etiological diagnosis of infectious diseases. Precise treatment can be carried out according to the positive mNGS results. However, the role of negative results of mNGS remains poorly defined in clinical practice. Methods: The results of 1,021 samples from patients who received the mNGS test at Zhongshan Hospital, Fudan University, between January 2019 and December 2019 were analyzed. Results: There were 308 samples (30.17%) of negative results included in the current study. The top 2 types of negative samples were blood (130/308) and tissue (63/308), which also accounted for the highest negative proportion in diseases. Sputum and bronchoalveolar lavage fluid (BALF) were more likely to have positive results. In false-negative results (defined as negative in mNGS test but reported positive in other sample types or assays), 118 samples were found when compared to regular microbiological assays. The negative predictive value (NPV) of mNGS was 95.79% [95%CI, 93.8%-97.8%] as compared to culture and smear. Mycobacterium, Aspergillus, and Mycoplasma ranked as the top 3 microorganisms on the undetected pathogen list. Conclusions: The present data indicate that when the mNGS test is negative, the negative prediction accuracy rate of the original specimen is significant. However, other laboratory assays results and clinical presentations should always be carefully considered prior to drawing a diagnosis.

Corrigendum: Research progress on microRNA-1258 in the development of human cancer.

[This corrects the article DOI: 10.3389/fonc.2022.1024234.].

Precision Breast-Conserving Surgery With Microwave Ablation Guidance: A Pilot Single-Center, Prospective Cohort Study.

INTRODUCTION: Negative margins in breast-conserving surgery (BCS) are essential for preventing recurrence. The aim of this study was to determine the use of preoperative microwave ablation (MWA) in the guidance of BCS for early-stage breast cancer and access whether MWA could influence the rates of positive resection margins. METHODS: From 2016 to 2018, 22 women with T1/T2 invasive breast cancer were enrolled for MWA prospectively in the guidance of BCS. US-guided MWA was performed under local anesthesia, followed by BCS and sentinel lymph node biopsy (SLNB) one week after ablation. Women who underwent palpation-guided BCS directly were included as control, and propensity score matching analysis was applied. RESULTS: MWA was performed in 22 patients. Of the 21 MWA cases with effect information, the mean tumor size in US was 20.9 ± 6.2 mm (6-37 mm). Compared with control group (BCS directly), a lower rate of positive/close margins was observed in MWA guidance group (P = 0.018), and MWA caused a higher rate of accurate surgery (the largest margin ≤ 3 cm and the smallest margin ≥ 1mm, P = 0.042). Of these 21 patients treated with MWA, 18 were candidates for SLNB. And sentinel lymph nodes were successfully identified in all cases, and no recurrence was found with a mean follow-up of 23 months. CONCLUSION: For patients with T1/T2 breast cancer, the application of preoperative MWA could guide BCS accurately without impairing SLNB. Clinical trials with long-term results are required to validate MWA in the guidance for breast cancer excision.

Microwave ablation induces Th1-type immune response with activation of ICOS pathway in early-stage breast cancer.

BACKGROUND: Despite great advances in the treatment of breast cancer, innovative approaches are still needed to reduce metastasis. As a minimally invasive local therapy (not standard therapy for breast cancer), microwave ablation (MWA) has been attempted to treat breast cancer, but the local effect and immune response induced by MWA have seldom been reported. METHODS: The clinical study was performed to determine the complete ablation rate of MWA for early-stage breast cancer. Secondary endpoints included safety and antitumor immune response. 35 subjects from this clinical study were enrolled in the current report, and the local effect was determined by pathological examinations or follow-up. To investigate MWA-induced immune response, patients treated with surgery (n=13) were enrolled as control, and blood samples were collected before and after MWA or surgery. The immune cell populations, serum cytokines, secretory immune checkpoint molecules, and T-cell receptor sequencing were analyzed. RESULTS: Of 35 enrolled patients, 32 (91.4%) showed complete ablation. Compared with surgery, MWA induced significantly increased levels of inducible co-stimulator (ICOS)+ activated CD4+ T cells and serum interferon gamma, indicating a shift in the Th1/Th2 balance toward Th1. The activated ICOS pathway was involved in the MWA-induced adaptive immune response. T-cell receptor sequencing revealed MWA of primary tumor activated T lymphocytes expansion and recognized some cancer-specific antigens. Moreover, CD4+ effector memory T-cell response was induced by MWA, and the immune response still existed after surgical resection of the ablated tumor. CONCLUSIONS: MWA may not only be a promising local therapy but also a trigger of antitumor immunity for breast cancer, opening new avenues for the treatment of breast cancer. Combinatorial strategy using additional agents which boost MWA-induced immune response could be considered as potential treatment for clinical study for early breast cancer therapy.

Comparison of Survival Outcomes Among Patients With Breast Cancer With Distant vs Ipsilateral Supraclavicular Lymph Node Metastases.

Importance: There is a lack of studies exploring whether the survival of patients with distant lymph node metastases (DLNM) is different from that of patients with ipsilateral supraclavicular lymph node metastases (ISLM) and other stage IV breast cancer. Objective: To assess the survival of patients with DLNM from breast cancer vs ISLM and other stage IV breast cancer. Design, Setting, and Participants: This cohort study included 2033 patients diagnosed with breast cancer between January 1, 2010, and December 31, 2014, from the Surveillance, Epidemiology and End Results registries database. Three groups of patients were included: (1) patients with ISLM without any distant metastasis, (2) patients with DLNM, and (3) patients with distant metastases (DLNM excluded). Patients younger than 18 years or older than 100 years were excluded. The data were analyzed in February 2020. Exposures: Surgery for primary tumor, surgery for distant lymph nodes, and radiotherapy. Main Outcomes and Measures: Overall survival (OS) and breast cancer-specific survival (BCSS). Results: Of the 2033 women (mean [SD] age, 62.03 [14.62] years [range, 23.00-99.00 years]; 1510 White participants [74.3%]) with breast cancer included in the study, 346 patients (17.0%) had DLNM, 212 (10.4%) had ISLM, and 1475 (72.6%) had distant metastases (DLNM excluded). The 3-year BCSS rates were 63.24% for ISLM, 64.54% for DLNM, and 41.20% for distant metastases. The 3-year OS rates were 53.46% for ISLM, 62.67% for DLNM, and 38.21% for distant metastases. Compared with patients with ISLM, patients with DLNM showed similar BCSS (hazard ratio [HR], 0.81; 95% CI, 0.52-1.25; P = .34) and OS (HR, 0.73; 95% CI, 0.51-1.05; P = .09), whereas patients with distant metastases showed significantly poorer BCSS (HR, 1.99; 95% CI, 1.43-2.78; P < .001) and OS (HR, 1.79; 95% CI, 1.35-2.38; P < .001). Of the 346 patients with DLNM, primary surgery (HR, 0.21; 95% CI, 0.12-0.39; P < .001) and radiotherapy (HR, 0.46; 95% CI, 0.25-0.87; P = .02) were significantly associated with improved OS. Conclusions and Relevance: The results of this cohort study suggest that DLNM of breast cancer, with similar survival to N3c disease (indicating metastases to the ipsilateral supraclavicular lymph nodes), might be a regional disease, and reassessment of the role of lymph node metastases in breast cancer may be necessary. Given these findings, aggressive locoregional therapies for this disease are recommended, although future studies are still needed to confirm these results.

Ultrasound Image Texture Feature Learning-Based Breast Cancer Benign and Malignant Classification.

The use of ultrasound images to acquire breast cancer diagnosis information without invasion can reduce the physical and psychological pain of breast cancer patients and is of great significance for the diagnosis and treatment of breast cancer. There are some differences in the texture of breast cancer between benign and malignant cases. Therefore, this paper proposes an adaptive learning method based on ultrasonic image texture features to identify breast cancer. Specifically, firstly, we used dictionary learning and sparse representation to learn the ultrasonic image texture dictionary of benign and malignant cases, respectively, and then used the combination of the two dictionaries to represent the test image to obtain the texture distribution characteristics of the test image under the two dictionary representations, which called the sparse representation coefficient. Finally, these above features were filtered by sparse representation and sent to sparse representation classifier to establish benign and malignant classification model. 128 cases were randomly divided into training and testing sets according to 2: 1 for training and testing. The proposed method has achieved state-of-the-art results, with an accuracy of 0.9070 and the area under the receiver operating characteristic curve of 0.9459. The results demonstrate that the proposed method has the potential to be used in the clinical diagnosis of benign and malignant breast cancer.

Microwave ablation of primary breast cancer inhibits metastatic progression in model mice via activation of natural killer cells.

Surgery is essential for controlling the symptoms and complications of stage IV breast cancer. However, locoregional treatment of primary tumors often results in distant progression, including lung metastasis, the most common type of visceral metastasis. As a minimally invasive thermal therapy, microwave ablation (MWA) has been attempted in the treatment of breast cancer, but the innate immune response after MWA has not yet been reported. Using two murine models of stage IV breast cancer, we found that MWA of primary breast cancer inhibited the progression of lung metastasis and improved survival. NK cells were activated after MWA of the primary tumor and exhibited enhanced cytotoxic functions, and the cytotoxic pathways of NK cells were activated. Depletion experiments showed that NK cells but not CD4+ or CD8+ T cells played a pivotal role in prolonging survival. Then, we found that compared with surgery or control treatment, MWA of the primary tumor induced completely different NK-cell-related cytokine profiles. Macrophages were activated after MWA of the primary tumor and produced IL-15 that activated NK cells to inhibit the progression of metastasis. In addition, MWA of human breast cancer stimulated an autologous NK-cell response. These results demonstrate that MWA of the primary tumor in metastatic breast cancer inhibits metastatic progression via the macrophage/IL-15/NK-cell axis. MWA of the primary tumor may be a promising treatment strategy for de novo stage IV breast cancer, although further substantiation is essential for clinical testing.

Multidisciplinary therapy strategy of precision medicine in clinical practice.

The application of precision medicine concept in clinical work needs a period of practice and experience accumulation. The present article introduced an example of functioning approach named "multidisciplinary therapy strategy of precision medicine" (MDTS-PM), clinical practice and process, decision-making, and therapies. The MDTS-PM integrates multidisciplinary experts and develops real-time therapeutic strategy based on clinical phenomes and gene sequencing of tissue DNA and circulating DNA. The strength of MDTS-PM is the combination of dynamical clinical phenomes, genetic information, diagnosis, and treatment to make the therapy more targeted and specific. MDTS-PM provides comprehensive, whole-process, and personalized diagnosis and treatment services for patients with complex cancer or complex drug resistance progression; provides guidance for further adjustment of drug use; and establishes a multidisciplinary cooperative team, improves the quality of clinical diagnosis and treatment, and optimizes the process of medical services.

ZFP57 suppress proliferation of breast cancer cells through down-regulation of MEST-mediated Wnt/ß-catenin signalling pathway.

Activation of oncogenes by promoter hypomethylation plays an important role in tumorigenesis. Zinc finger protein 57 (ZFP57), a member of KRAB-ZFPs, could maintain DNA methylation in embryonic stem cells (ESCs), although its role and underlying mechanisms in breast cancer are not well understood. In this study, we found that ZFP57 had low expression in breast cancer, and overexpression of ZFP57 could inhibit the proliferation of breast cancer cells by inhibiting the Wnt/ß-catenin pathway. MEST was validated as the direct target gene of ZFP57 and MEST may be down-regulated by ZFP57 through conserving DNA methylation. Furthermore, overexpression of MEST could restore the tumour-suppressed and the Wnt/ß-catenin pathway inactivated effects of ZFP57. ZFP57-MEST and the Wnt/ß-catenin pathway axis are involved in breast tumorigenesis, which may represent a potential diagnostic biomarker, and provide a new insight into a novel therapeutic strategy for breast cancer patients.

The methodology study of three-dimensional (3D) genome research.

Multiple dimensions of genome organization play critical roles in the regulation of non-coding regions in gene expression in cell/organ development and pathogenesis. Precise measurements of multi-dimensional genome structure ensure data quality and fully depend upon the study design. We here overview the number of methodologies used in the detection and analysis of genome structure and compare advantages and disadvantages of 3C-based, PCR amplification-based, and sequencing-based measurements. We discuss about the optimization of various techniques according to targeted genomic sites, the required resolution, and possible technique biases. Comparison of different analysis tools and computational system-based automatic analysis is evaluated to define more opportunities and challenges of data analysis in 3D genome research. The genome structure is visualized in levels of single genome organized by enhancer-promoter interactions, TAD, and intra-chromosomal and inter-chromosomal interactions between TADs. Thus, methodologies of genome organization multi-dimensions are not only critical in studies on 3D genome-regulated transcriptions, but also in the discovery of disease-specific biomarkers and targets for diagnosis and therapies.

Concordance assessment of Watson for Oncology in breast cancer chemotherapy: first China experience.

BACKGROUND: IBM Watson for Oncology (WFO) is an artificial intelligence cognitive computing system that provides confidence-ranked, evidence-based treatment recommendations for cancer. We examine the level of agreement for breast cancer chemotherapy between WFO recommended and clinical use in a large population of breast cancer cases. METHODS: A total of 1,301 breast cancer patients were reviewed in The First Affiliated Hospital with Nanjing Medical University, China from June 2013 to December 2017. Patients' data were entered manually into WFO by the trained senior oncology fellows. Chemotherapy recommendations were provided in 3 categories, "Recommended", "For Consideration", and "Not Recommended". Concordance was achieved when oncologists' treatment suggestions were in the "Recommended" or "For Consideration" categories. RESULTS: The chemotherapy regimen concordance was 69.4% among all breast cancer cases, 65.0% among the cases in adjuvant chemotherapy (AC) group and 96.7% among the cases in neoadjuvant chemotherapy (NAC) group. The concordance varied greatly in subset analysis with respect to TNM stage and molecular subtype. AC recommendations were concordant in 92.3% of stage III breast cancer and 50.8% of stage I. However, the concordance varied by molecular subtype, which was higher for triple negative breast cancer (89.3%) than others. The chemotherapy regimen concordance declined significantly with increasing age, except for the age group 41-50 years. CONCLUSIONS: Chemotherapy regimens provided by WFO did not exhibit a high degree of agreement with those suggested by oncologists in clinical practice in the hospital in China. The current effort is underway to enhance WFO's capabilities as a cognitive decision support tool by incorporating regional guidelines, enabling oncologists and patients to benefit from WFO worldwide.

Detection and Application of RNA Editing in Cancer.

RNA editing is the process which happened in the post-transcriptional stage that the genetic information contained in an RNA molecule will be changed. RNA editing has been found to be related with many cancers, so through identifying RNA editing sites, we can find useful information on the process of carcinogenesis. In this review, we will discuss the main types of RNA editing and their role in cancers, as well as the current detection methods of RNA editing and the challenges we should overcome.

Approach, Application, and Bioethics of mtDNA Sequencing in Cancer.

Mitochondrial DNA (mtDNA) is more vulnerable to mutations and associated with many solid tumors. Through mtDNA sequencing, we can find useful information on the mutations implicated in diseases and can better define the impact of mitochondrial dysfunction on the process of carcinogenesis. In current article, we will discuss the current approaches of mtDNA sequencing and the challenges we should overcome, their applications in various cancers, and the potential bioethics problems we should face in the application of mtDNA sequencing in clinical diagnosis and treatment.