CD248-expressing cancer-associated fibroblasts induce non-small cell lung cancer metastasis via Hippo pathway-mediated extracellular matrix stiffness.

Metastasis is a crucial stage in tumour progression, and cancer-associated fibroblasts (CAFs) support metastasis through their participation in extracellular matrix (ECM) stiffness. CD248 is a possible biomarker for non-small cell lung cancer (NSCLC)-derived CAFs, but its role in mediating ECM stiffness to promote NSCLC metastasis is unknown. We investigated the significance of CD248+ CAFs in activating the Hippo axis and promoting connective tissue growth factor (CTGF) expression, which affects the stromal collagen I environment and improves ECM stiffness, thereby facilitating NSCLC metastasis. In this study, we found that higher levels of CD248 in CAFs induced the formation of collagen I, which in turn increased extracellular matrix stiffness, thereby enabling NSCLC cell infiltration and migration. Hippo axis activation by CD248+ CAFs induces CTGF expression, which facilitates the formation of the collagen I milieu in the stromal matrix. In a tumour lung metastasis model utilizing fibroblast-specific CD248 gene knockout mice, CD248 gene knockout mice showed a significantly reduced ability to develop tumour lung metastasis compared to that of WT mice. Our findings demonstrate that CD248+ CAFs activate the Hippo pathway, thereby inducing CTGF expression, which in turn facilitates the collagen I milieu of the stromal matrix, which promotes NSCLC metastasis.

A Comparative Study of Antitumor Immunity Induced by Radiofrequency Microwave and Cryoablation in Hepatocellular Carcinoma.

PURPOSE: This study aimed to compare the immune responses induced by microwave ablation (MWA), radiofrequency ablation (RFA), and cryoablation (CRYO) in hepatocellular carcinoma (HCC) and identify differences in immune responses and the timing of immune changes. MATERIALS AND METHODS: A bilateral subcutaneous model was established in C57 mice, and the successfully modeled mice were divided into the microwave (n = 15), radiofrequency (n = 15), CRYO (n = 15), control (n = 9), and blank groups (n = 3). Mice in the control group were dissected before ablation, whereas mice in the three ablation groups underwent ultrasound-guided ablation of one axillary tumor. Three mice were sacrificed and dissected at 1-4 weeks after ablation. After tissue processing, flow cytometry was used to detect the levels of CD8 + T and regulatory T (Treg) cells in the tissue, and western blotting was used to assess the level of programmed cell death ligand 1 (PD-L1) protein in the tumor tissue. RESULTS: The pattern of immune changes after the three types of ablation was consistent, with immune changes occurring at 3-4 weeks. CRYO induced the most significant increase in the percentage of CD8 + T cells. There were no significant differences in the levels of Treg cells and the level of PD-L1 protein among the three types of ablation (p > 0.05), but the decline in Treg cells and PD-L1 protein level caused by CRYO was the most pronounced. CONCLUSION: In the HCC mouse model, the immune changes following the three types of ablation were consistent, with immune changes occurring at 3-4 weeks. Among them, CRYO elicited the strongest adaptive immune response, and RFA outperformed MWA.

Prevalence and clinico-genomic characteristics of patients with TRK fusion cancer in China.

Neurotrophic tyrosine kinase (NTRK) fusions involving NTRK1, NTRK2, and NTRK3 were found in a broad range of solid tumors as driver gene variants. However, the prevalence of NTRK fusions in Chinese solid tumor patients is rarely reported. Based on the next-generation sequencing data from 10,194 Chinese solid tumor patients, we identified approximately 0.4% (40/10,194) of Chinese solid tumor patients with NTRK fusion. NTRK fusions were most frequently detected in soft tissue sarcoma (3.0%), especially in the fibrosarcoma subtype (12.7%). A total of 29 NTRK fusion patterns were identified, of which 11 were rarely reported. NTRK fusion mostly co-occurred with TP53 (38%), CDKN2A (23%), and ACVR2A (18%) and rarely with NTRK amplification (5.0%) and single nucleotide variants (2.5%). DNA-based NTRK fusion sequencing exhibited a higher detection rate than pan-TRK immunohistochemistry (100% vs. 87.5%). Two patients with NTRK fusions showed clinical responses to larotrectinib, supporting the effective response of NTRK fusion patients to TRK inhibitors.

Three-dimensional template combined with MR-guided iodine-125 brachytherapy for recurrent brain metastases.

Purpose: Treatment of recurrent brain metastases is extremely challenging. Here, we evaluated the feasibility and efficacy of an individualized three-dimensional template combined with MR-guided iodine-125 (125I) brachytherapy in the treatment of recurrent brain metastases. Material and methods: Twenty-eight patients with recurrent 38 brain metastases underwent 125I brachytherapy from December, 2017 to January, 2021. A pre-treatment brachytherapy plan and three-dimensional template were generated according to isovoxel T1-weighted MR images. 125I seeds were implanted under the guidance of three-dimensional template and 1.0-T open MR imaging. Dosimetry verification was performed based on CT/MR fusion images. Pre-operative and post-operative dosimetry parameters of D90, V100, and conformity index (CI) were compared. Overall response rate (ORR), disease control rate (DCR) at 6 months, and 1-year survival rate were calculated. Median overall survival (OS) measured from the date of 125I brachytherapy was estimated using Kaplan-Meier method. Results: No significant differences were observed between pre-operative and post-operative D90, V100, and CI values (p > 0.05). The ORR and DCR at 6 months were 91.3% and 95.7%, respectively. The 1-year survival rate was 57.1%. The median OS was 14.1 months. Two cases of minor hemorrhage and 5 cases of symptomatic brain edema were observed during the study. All clinical symptoms were alleviated after corticosteroid treatment applied for 7 to 14 days. Conclusions: A three-dimensional template combined with MR-guided 125I brachytherapy in the treatment of recurrent brain metastases is feasible, safe, and effective. This novel 125I brachytherapy strategy is an attractive alternative in the treatment of brain metastases.

Regional hyperthermia with cisplatin added to gemcitabine versus gemcitabine in patients with resected pancreatic ductal adenocarcinoma: The HEAT randomised clinical trial.

BACKGROUND: Regional hyperthermia (RHT) with cisplatin added to gemcitabine showed efficacy in gemcitabine-pre-treated patients with advanced pancreatic ductal adenocarcinoma. We conducted a randomised clinical trial to investigate RHT with cisplatin added to gemcitabine (GPH) compared with gemcitabine (G) in the adjuvant setting of resected pancreatic ductal adenocarcinoma. METHODS: This randomised, multicentre, open-label trial randomly assigned patients to either GPH (gemcitabine 1000 mg/m2 on day 1, 15 and cisplatin 25 mg/m2 with RHT on day 2, 3 and 15,16) or to G (gemcitabine 1000 mg/m2 on day 1,8,15), four-weekly over six cycles. Disease-free survival (DFS) was the primary end-point. Secondary end-points included overall survival (OS) and safety. RESULTS: A total of 117 eligible patients (median age, 63 years) were randomly allocated to treatment (57 GPH; 60 G). With a follow-up time of 56.6 months, the median DFS was 12.7 compared to 11.2 months for GPH and G, respectively (p = 0.394). Median post-recurrence survival was significantly prolonged in the GPH-group (15.3 versus 9.8 months; p = 0.031). Median OS reached 33.2 versus 25.2 months (p = 0.099) with 5-year survival rates of 28.4% versus 18.7%. Excluding eight patients who received additional capecitabine in the G-arm (investigators choice), median OS favoured GPH (p = 0.052). Adverse events CTCAE (Common Terminology Criteria for Adverse Events) grade ≥3 occurred in 61.5% (GPH) versus 63.6% (G) of patients. Two patients in the G-group died because of treatment-related toxic effects. CONCLUSIONS: The randomised controlled Hyperthermia European Adjuvant Trial study failed to demonstrate a significant difference in DFS. However, it suggests a difference in post-recurrence survival and a trend for improved OS. CLINICALTRIALS: gov, number NCT01077427.

An in-library ligation strategy and its application in CRISPR/Cas9 screening of high-order gRNA combinations.

Simultaneous targeting multiple genes is a big advantage of CRISPR (clustered regularly interspaced short palindromic repeats) genome editing but challenging to achieve in CRISPR screening. The crosstalk among genes or gene products is a common and fundamental mechanism to ensure cellular stability and functional diversity. However, the screening approach to map high-order gene combinations to the interesting phenotype is still lacking. Here, we developed a universal in-library ligation strategy and applied it to generate multiplexed CRISPR library, which could perturb four pre-designed targets in a cell. We conducted in vivo CRISPR screening for potential guide RNA (gRNA) combinations inducing anti-tumor immune responses. Simultaneously disturbing a combination of three checkpoints in CD8+ T cells was demonstrated to be more effective than disturbing Pdcd1 only for T cell activation in the tumor environment. This study developed a novel in-library ligation strategy to facilitate the multiplexed CRISPR screening, which could extend our ability to explore the combinatorial outcomes from coordinated gene behaviors.

Efficacy of using tidal volume challenge to improve the reliability of pulse pressure variation reduced in low tidal volume ventilated critically ill patients with decreased respiratory system compliance.

BACKGROUND: The prediction accuracy of pulse pressure variation (PPV) for fluid responsiveness was proposed to be unreliable in low tidal volume (Vt) ventilation. It was suggested that changes in PPV obtained by transiently increasing Vt to 8 ml/kg accurately predicted fluid responsiveness even in subjects receiving low Vt. We assessed whether the changes in PPV induced by a Vt challenge predicted fluid responsiveness in our critically ill subjects ventilated with low Vt 6 ml/kg. METHODS: This study is a prospective single-center study. PPV and other parameters were measured at a Vt of 6 mL/kg, 8 mL/kg, and after volume expansion. The prediction accuracy of PPV and other parameters for fluid responsiveness before and after tidal volume challenge was also analyzed using receiver operating characteristic (ROC) curves. RESULTS: Thirty-one of the 76 subjects enrolled in the study were responders (41%). Respiratory system compliance of all subjects decreased significantly (26 ± 4.3). The PPV values were significantly higher in the responder group than the non-responder group before (8.8 ± 2.7 vs 6.8 ± 3.1) or after (13.0 ± 1.7 vs 8.5 ± 3.0) Vt challenge. In the receiver operating characteristic curve (ROC) analysis, PPV6 showed unsatisfactory predictive capability with an area under the curve (AUC) of 0.69 (95%CI, 0.57-0.79, p = 0.002) at a Vt of 6 mL/kg. PPV8 andΔPPV6-8 showed good predictive capability with an AUC of 0.90 (95% CI, 0.81-0.96, p < 0.001) and 0.90 (95% CI, 0.80-0.95, P < 0.001) respectively. The corresponding cutoff values were 11% for PPV8 and 2% for ΔPPV6-8. CONCLUSIONS: PPV shows a poor operative performance as a predictor of fluid responsiveness in critically ill subjects ventilated with a tidal volume of 6 mL/ kg. Vt challenge could improve the predictive accuracy of PPV to a good but not excellent extent when respiratory system compliance decreased significantly.

Validating the knowledge bank approach for personalized prediction of survival in acute myeloid leukemia: a reproducibility study.

Reproducibility is not only essential for the integrity of scientific research but is also a prerequisite for model validation and refinement for the future application of predictive algorithms. However, reproducible research is becoming increasingly challenging, particularly in high-dimensional genomic data analyses with complex statistical or algorithmic techniques. Given that there are no mandatory requirements in most biomedical and statistical journals to provide the original data, analytical source code, or other relevant materials for publication, accessibility to these supplements naturally suggests a greater credibility of the published work. In this study, we performed a reproducibility assessment of the notable paper by Gerstung et al. (Nat Genet 49:332-340, 2017) by rerunning the analysis using their original code and data, which are publicly accessible. Despite an open science setting, it was challenging to reproduce the entire research project; reasons included: incomplete data and documentation, suboptimal code readability, coding errors, limited portability of intensive computing performed on a specific platform, and an R computing environment that could no longer be re-established. We learn that the availability of code and data does not guarantee transparency and reproducibility of a study; paradoxically, the source code is still liable to error and obsolescence, essentially due to methodological and computational complexity, a lack of reproducibility checking at submission, and updates for software and operating environment. The complex code may also hide problematic methodological aspects of the proposed research. Building on the experience gained, we discuss the best programming and software engineering practices that could have been employed to improve reproducibility, and propose practical criteria for the conduct and reporting of reproducibility studies for future researchers.

SPION-MSCs enhance therapeutic efficacy in sepsis by regulating MSC-expressed TRAF1-dependent macrophage polarization.

BACKGROUND: Sepsis is defined as a life-threatening organ dysfunction caused by a dysregulated host response to infection. The liver has a crucial role in sepsis and is also a target for sepsis-related injury. Macrophage polarization between the M1 and M2 types is involved in the progression and resolution of both inflammation and liver injury. Iron oxide-based synthetic nanoparticles (SPIONs) can be used as antibacterial agents to regulate the inflammatory response. Mesenchymal stromal/stem cells (MSCs) have been widely used in the treatment of autoimmune diseases, sepsis, and other diseases. However, to date, both the effects of SPIONs on MSCs and the fate of SPION-labelled MSCs in sepsis and other diseases are still unclear. METHODS: Mice were subjected to caecal ligation and puncture (CLP) or lipopolysaccharide (LPS) induction to develop sepsis models. The CLP or LPS models were treated with MSCs or SPION-labelled/pretreated MSCs (SPION-MSCs). Bone marrow (BM)-derived macrophages and RAW 264.7 cells were cocultured with MSCs or SPION-MSCs under different conditions. Flow cytometry, transmission electron microscopy, western blotting, quantitative real-time PCR, and immunohistochemical analysis were performed. RESULTS: We found that SPIONs did not affect the basic characteristics of MSCs. SPIONs promoted the survival of MSCs by upregulating HO-1 expression under inflammatory conditions. SPION-MSCs enhanced the therapeutic efficacy of liver injury in both the CLP- and LPS-induced mouse models of sepsis. Moreover, the protective effect of SPION-MSCs against sepsis-induced liver injury was related to macrophages. Systemic depletion of macrophages reduced the efficacy of SPION-MSC therapy. Furthermore, SPION-MSCs promoted macrophages to polarize towards the M2 phenotype under sepsis-induced liver injury in mice. The enhanced polarization towards M2 macrophages was attributed to their phagocytosis of SPION-MSCs. SPION-MSC-expressed TRAF1 was critical for promotion of macrophage polarization and alleviation of sepsis in mice. CONCLUSION: MSCs labelled/pretreated with SPIONs may be a novel therapeutic strategy to prevent or treat sepsis and sepsis-induced liver injury. HIGHLIGHTS: 1. SPIONs enhance the viability of MSCs by promoting HO-1 expression. 2. SPION-labelled/pretreated MSCs effectively improve sepsis by regulating macrophage polarization to M2 macrophages. 3. SPION-labelled/pretreated MSCs regulate macrophage polarization in a manner dependent on MSC-expressed TRAF1 protein.

Immune infiltrates in patients with localised high-risk soft tissue sarcoma treated with neoadjuvant chemotherapy without or with regional hyperthermia: A translational research program of the EORTC 62961-ESHO 95 randomised clinical trial.

BACKGROUND: The EORTC 62961-ESHO 95 randomised trial showed improved long-term survival of patients with high-risk soft-tissue sarcoma by adding regional hyperthermia to neoadjuvant chemotherapy. We hypothesised that immune infiltrate of patients treated with neoadjuvant therapy associate with clinical outcome. METHODS: Tumour infiltrating lymphocytes (TILs) and CD8, FOXP3, PD-1, and PD-L1 were evaluated in sequential biopsies of patients after four cycles of therapy. RESULTS: From a subgroup of 109 patients who had been randomised between July 1997 and November 2006 to neoadjuvant chemotherapy (53 patients) or neoadjuvant chemotherapy with regional hyperthermia (56 patients), 137 biopsies were obtained. TILs increased in paired second biopsies independent of treatment allocation (p < 0.001). FOXP3 regulatory T cells decreased (p = 0.002), and PD-L1 expression of tumours became undetectable. In the multivariate analysis, post-treatment high TILs correlated to LPFS (HR: 0.34; 95% CI 0.15-0.75; p = 0.008) and DFS (HR: 0.38; 95% CI 0.17-0.82; p = 0.015). In comparing post-treatment immune infiltrate between treatment arms, tumour response was associated with neoadjuvant chemotherapy with regional hyperthermia (p = 0.013) and high TILs (p = 0.064). High CD8 cell infiltration was associated with improved LPFS (HR: 0.27; 95% CI 0.09-0.79; Log-rank p = 0.011) and DFS (HR: 0.25; 95% CI 0.09-0.73; Log-rank p = 0.006). Improved survival at 10 years was associated with immune infiltrate after neoadjuvant chemotherapy with regional hyperthermia. CONCLUSION: Preoperative therapy re-programs a non-inflamed tumour at baseline into an inflamed tumour. The post-treatment immune infiltrate became predictive for clinical outcomes. The combination with regional hyperthermia primes the tumour microenvironment, enabling enhanced anti-tumour immune activity in high-risk soft tissue sarcomas. TRIAL REGISTRATION: ClinicalTrials.gov, NCT00003052.

Chitosan-Poly(Acrylic Acid) Nanoparticles Loaded with R848 and MnCl2 Inhibit Melanoma via Regulating Macrophage Polarization and Dendritic Cell Maturation.

PURPOSE: Since immune cells in the tumor microenvironment (TME) can affect the development and progression of tumors, strategies modulating immune cells are considered to have an important therapeutic effect. As a TLR7/8 agonist, R848 effectively activates the innate immune cells to exert an anti-tumor effect. Mn2+ has been reported to strongly promote the maturation of antigen-presenting cells (APCs), thereby enhancing the cytotoxicity of CD8+ T cells. Thus, we tried to investigate whether chitosan-poly(acrylic acid) nanoparticles (CS-PAA NPs) loaded with R848 and MnCl2 (R-M@CS-PAA NPs) could exert an anti-tumor effect by regulating the function of immune cells. METHODS: R-M@CS-PAA NPs were prepared, and their basic characteristics, anti-tumor effect, and potential mechanisms were explored both in vitro and in vivo. RESULTS: R-M@CS-PAA NPs easily released MnCl2 and R848 at low pH. In B16F10 mouse melanoma model, R-M@CS-PAA NPs exerted the most significant anti-melanoma effect compared with the control group and CS-PAA NPs loaded with R848 or MnCl2 alone. FITC-labeled R-M@CS-PAA NPs were displayed to be accumulated at the tumor site. R-M@CS-PAA NPs significantly increased the infiltration of M1 macrophages and CD8+ T cells but reduced the number of suppressive immune cells in the TME. Moreover, in vitro experiments showed that R-M@CS-PAA NPs polarized macrophages into the M1 phenotype to inhibit the proliferation of B16F10 cells. R-M@CS-PAA NPs also enhanced the killing function of CD8+ T cells to B16F10 cells. Of note, R-M@CS-PAA NPs not only promoted the maturation of APCs such as dendritic cells and macrophages by STING and NF-кB pathways, but also enhanced the ability of dendritic cells to present ovalbumin to OT-I CD8+ T cells to enhance the cytotoxicity of OT-I CD8+ T cells to ovalbumin-expressing B16F10 cells. CONCLUSION: These data indicate that the administration of R-M@CS-PAA NPs is an effective therapeutic strategy against melanoma.

Ferumoxytol-ß-glucan Inhibits Melanoma Growth via Interacting with Dectin-1 to Polarize Macrophages into M1 Phenotype.

Background: Regulating the polarization of macrophages to antitumor M1 macrophages is a promising strategy for overcoming the immunosuppression of the tumor microenvironment for cancer therapy. Ferumoxytol (FMT) can not only serve as a drug deliver agent but also exerts anti-tumor activity. ß-glucan has immuno-modulating properties to prevent tumor growth. Thus, a nanocomposite of FMT surface-coated with ß-glucan (FMT-ß-glucan) was prepared to explore its effect on tumor suppression. Methods: Male B16F10 melanoma mouse model was established to explore the antitumor effect of FMT-ß-glucan. The viability and apoptotic rates of B16F10 cells were detected by cell counting kit-8 and Annexin-V/PI experiments. The levels of M1 markers were quantified by quantitative reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assay. Phagocytic activity and intracellular reactive oxygen species (ROS) in macrophages were evaluated by the neutral red uptake assay and flow cytometry, respectively. Small interfering RNA (siRNA) transfection was applied to knock down the Dectin-1 gene in RAW 264.7 cells. Results: FMT-ß-glucan suppressed tumor growth to a greater extent and induced higher infiltration of M1 macrophages than the combination of FMT and ß-glucan (FMT+ß-glucan) in vivo. In vitro, supernatant from FMT-ß-glucan-treated RAW 264.7 cells led to lower cell viability and induced more apoptosis of B16F10 cells than that from the FMT+ß-glucan group. Moreover, FMT-ß-glucan boosted the expression of M1 type markers, and increased phagocytic activity and ROS in RAW 264.7 cells. Further research indicated that FMT-ß-glucan treatment promoted the level of Dectin-1 on the surface of RAW 264.7 cells and that knockdown of Dectin-1 abrogated the phosphorylation levels of several components in MAPK and NF-κB signaling. Conclusion: The nanocomposite FMT-ß-glucan suppressed melanoma growth by inducing the M1 macrophage-activated tumor microenvironment.

Magnetic resonance-guided repeat biopsy of suspicious malignant lung lesions after an initial negative computed tomography-guided Biopsy.

Objective: This study sought to establish the diagnostic utility of performing a second biopsy using an magnetic resonance (MR)-guided percutaneous transthoracic needle biopsy (PTNB) approach in patients with suspicious malignant lung lesions that had already undergone an initial negative computed tomography (CT)-guided biopsy. Materials and Methods: This study evaluated 31 patients with suspicious lung lesions (18 males, 13 females; mean age: 62.1 ± 11.3 years) that had previously undergone CT-guided PTNB with negative pathological findings January 2015-November 2020. A final histopathological diagnosis was made based on resected lung lesion specimens or, when resection was not conducted, on clinical diagnosis following a ≥6-month follow-up. The diagnostic accuracy of MR-guided secondary lung biopsy was determined by comparing the lung biopsy results for each patient to their final diagnosis. Results: 1.0T open MR-guided secondary lung biopsy was performed for 31 lesions (20 central, 11 peripheral; mean size, 5.3 ± 2.0 cm). The pathological results revealed 20/31 (64.5%) lesions to be malignant (14 adenocarcinoma, 4 squamous cell carcinoma, and 2 small-cell lung cancer) as detected by 1.0T open MR-guided PTNB and confirmed by surgical pathology and clinical follow-up. There were three instances of biopsy-induced complications including hemorrhage in 6.5% of the patients (2/31) and pneumothorax in 3.2% of the patients (1/31). No patients experienced severe complications. Conclusion: For individuals with clinically suspicious lung lesions that initially received negative CT-guided PTNB findings, 1.0T open MR-guided secondary lung biopsy is a safe and effective secondary diagnostic approach.

Magnetic resonance-guided ablation of liver tumors: A systematic review and pooled analysis.

PURPOSE: The purpose of this study is to study the clinical outcomes of different types of magnetic resonance (MR)-guided ablation for the treatment of liver tumors by performing a systematic review and pooled analysis. MATERIALS AND METHODS: A comprehensive literature search was performed for clinical trials published from January 1997 to October 2019 in PubMed, the Web of Science, Embase, and the Cochrane Library. Pooled analyses were performed to obtain the complete ablation (CA), complication, progression-free survival (PFS), and overall survival (OS) rates. RESULTS: Thirty studies were eligible, including four studies on MR-guided microwave ablation (MWA); 14 studies on MR-guided radiofrequency ablation (RFA); one study on both MR-guided MWA and RFA; eight studies on MR-guided, laser-induced thermotherapy (LITT); two studies on MR-guided percutaneous cryoablation (PC); and one study on MR-guided percutaneous ethanol injection (PEI). The CA rates in patients who underwent RFA, MWA, LITT, PC, and PEI were 95.60%, 98.86%, 77.78%, 47.92%, and 85.71%, respectively. The most frequent complications were pain (27.66%, 13/47) and postablation syndrome (27.66%, 13/47) in the PC group; pleural effusion (8.11%, 119/1,468) and subcapsular hematoma (2.25%, 33/1,468) in the LITT group; pleural effusion (2.67%, 2/75) in the MWA group; and subcapsular hematoma (4.18%, 20/478) and post-ablation syndrome (2.93%, 14/478) in the RFA group. There were few studies reporting PFS and OS. CONCLUSIONS: MR-guided ablation is a practicable alternative treatment for liver tumors, especially MR-guided RFA and MWA, which have high rates of CA and low occurrences of complications.

17ß-Estradiol Promotes Trained Immunity in Females Against Sepsis via Regulating Nucleus Translocation of RelB.

Sepsis is more common among males than females, and the unequal estrogen levels have been suspected to play a vital role in gender differences. Recently, trained immunity is reported to be a novel strategy for the innate immune system to fight infection. However, it has not been clarified whether ß-glucan-induced trained immunity causes different responses to early sepsis between male and female mice. In this study, sepsis was induced in mice by intraperitoneal injection of Escherichia coli (E. coli). The changes of inflammatory cytokines expression, and macrophage polarization in male, female, and ovariectomized C57BL/6 mice in sepsis model were investigated. For in vitro studies, different macrophages were treated with LPS. The function of estradiol (E2) on macrophage cell lines was verified and the mechanism of E2 affecting trained immunity was explored. We demonstrated that ß-glucan-induced trained immunity was more resistant to sepsis in female than male mice. Macrophage polarization toward the M1 phenotype, which exhibited enhanced trained immunity, was related to the difference in sepsis resistance between female and male mice. Moreover, ovariectomized (OVX) mice manifested serious sepsis consequences with a weaker trained immunity effect than female mice. Female bone marrow-derived macrophages (BMDMs) were also apt to be polarized to the M1 phenotype in response to trained immunity in vitro. Furthermore, E2 promoted trained immunity in macrophage cell lines J774 and RAW264.7. E2 was also verified to facilitate trained immunity in primary BMDMs from female and male mice. Mechanistically, we found that E2 inhibited the nuclear translocation of RelB, which is a member of non-canonical pathway of NFκB and contributes to macrophage polarization to change the intensity of trained immunity. This study is the first to indicate the role of E2 in the trained immunity induced by ß-glucan to protect against E. coli-induced sepsis via the non-canonical NFκB pathway. These results improve our understanding of the molecular mechanisms governing trained immunity in gender differences.

Ganoderma lucidum fruiting body extracts inhibit colorectal cancer by inducing apoptosis, autophagy, and G0/G1 phase cell cycle arrest in vitro and in vivo.

Although previous studies have found that Ganoderma lucidum extracts have the ability to directly resist tumor proliferation and reduce metastasis and invasion, the effect of the extracts of Ganoderma lucidum fruiting body (GLE) on cancer is not clarified. This study intends to investigate the anticancer role of GLE on HCT116 colorectal cancer cells in vitro and in vivo. The effects of GLE on the proliferation, apoptosis, autophagy and cell cycle arrest of HCT116 cells were detected by cell counting kit-8 (CCK-8), flow cytometry, electron microscope, quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assay. Xenografted mouse models were used to evaluate the tumor growth inhibition effect of GLE in vivo. GLE could significantly inhibit the viability of four tumor cell lines (A549, SW1990, SKOV3 and HCT116) and HCT116 cells were more sensitive to GLE treatment with a half inhibitory concentration of 106 µg/mL. GLE treatment induced apoptosis of HCT116 cells by downregulating of the ratio of Bcl-2 to Bax and increasing cleaved caspase-3 and poly ADP-ribose polymerase (PARP) protein expression. Autophagy of HCT116 cells also increased after GLE treatment, as shown by observation of autophagosomes formation and altered protein expressions in the mTOR pathway. In addition, GLE treatment led to G0/G1 cell cycle arrest as evidenced by flow cytometry analysis and changes in cell-cycle-related gene expressions at the mRNA levels. Of note, in vivo evaluation indicated that GLE significantly inhibited tumor weight and tumor volume and decreased Ki67 expression. In summary, GLE has potential to be developed as an anticancer agent against colorectal cancer, and further evaluation is needed.

A novel three-dimensional template combined with MR-guided 125I brachytherapy for recurrent glioblastoma.

BACKGROUND: At present, the treatment of recurrent glioblastoma is extremely challenging. In this study, we used a novel three-dimensional non-coplanar template (3DNPT) combined with open MR to guide 125I seed implantation for recurrent glioblastoma. The aim of this study was to evaluate the feasibility, accuracy, and effectiveness of this technique. METHODS: Twenty-four patients of recurrent glioblastoma underwent 3DNPT with open MR-guided 125I brachytherapy from August 2017 to January 2019. Preoperative treatment plan and 3DNPT were made according to enhanced isovoxel T1-weighted MR images. 125I seeds were implanted using 3DNPT and 1.0-T open MR imaging guidance. Dosimetry verification was performed after brachytherapy based on postoperative CT/MR fusion images. Preoperative and postoperative dosimetry parameters of D90, V100, V200, conformity index (CI), external index (EI) were compared. The objective response rate (ORR) at 6 months and 1-year survival rate were calculated. Median overall survival (OS) measured from the date of brachytherapy was estimated by Kaplan-Meier method. RESULTS: There were no significant differences between preoperative and postoperative dosimetry parameters of D90, V100, V200, CI, EI (P > 0.05). The ORR at 6 months was 75.0%. The 1-year survival rate was 58.3%. Median OS was 12.9 months. One case of small amount of epidural hemorrhage occurred during the procedure. There were 3 cases of symptomatic brain edema after brachytherapy treatment, including grade three toxicity in 1 case and grade two toxicity in 2 cases. The three patients were treated with corticosteroid for 2 to 4 weeks. The clinical symptoms related to brain edema were significantly alleviated thereafter. CONCLUSIONS: 3DNPT combined with open MR-guided 125I brachytherapy for circumscribed recurrent glioblastoma is feasible, effective, and with low risk of complications. Postoperative dosimetry matched the preoperative treatment plan. The described method can be used as a novel implantation technique for 125I brachytherapy in the treatment of recurrent gliomas. TRIAL REGISTRATION: The study was approved by the Institutional Review Board of Shandong Provincial Hospital Affiliated to Shandong University (NSFC:NO.2017-058), registered 1st July 2017.

Randomized Comparison of Intensified and Standard P2Y12-Receptor-Inhibition Before Elective Percutaneous Coronary Intervention: The SASSICAIA Trial.

BACKGROUND: Even among biomarker-negative patients undergoing elective percutaneous coronary intervention (PCI), periprocedural thrombotic and bleeding complications can lead to increased morbidity and mortality. Whether stronger platelet inhibition by an intensified oral loading strategy (ILS) before PCI impacts on outcomes among these patients in contemporary practice remains unclear. METHODS: This multicenter, randomized, assessor-blinded trial tested the hypothesis that in elective PCI prasugrel 60 mg (ILS) is superior to standard loading strategy with clopidogrel 600 mg regarding a composite primary end point of all-cause death, any myocardial infarction, definite/probable stent thrombosis, stroke, or urgent vessel revascularization. After PCI, all patients were on clopidogrel 75 mg/day and aspirin. The trial was terminated prematurely because of slower-than-expected recruitment and funding discontinuation. RESULTS: Of 781 patients included in the final analysis, 382 were assigned to ILS and 399 to standard loading strategy. At 30 days, the primary end point occurred in 66 patients (17.3%) assigned to ILS and 74 patients (18.6%) assigned to standard loading strategy (odds ratio, 0.92 [95% CI, 0.63-1.32]; P=0.64). Any myocardial infarction and Bleeding Academic Research Consortium ≥2 bleeding rates were similar among ILS and standard loading strategy groups 16.2% versus 17.5%, odds ratio, 0.91 (95% CI, 0.62-1.32), P=0.62 and 4.2% versus 4.8%, odds ratio 0.87 (95% CI, 0.44-1.73), P=0.70, respectively. CONCLUSIONS: In biomarker-negative stable and unstable angina patients undergoing elective PCI, the trial did not find a conclusive difference in efficacy or safety. This observation should be interpreted in the context of wide CIs and premature termination of the trial. Registration: URL: https://www.clinicaltrials.gov. Unique identifier: NCT02548611.

Multiparametric magnetic resonance-guided and monitored microwave ablation in liver cancer.

PURPOSE: The objective of our study was to prospectively evaluate the feasibility, effectiveness, and safety of 1.0T open multiparametric magnetic resonance (MR)-guided and monitored microwave ablation (MWA) of liver cancer. MATERIALS AND METHODS: Fifty-six liver lesions (12 - initial hepatocellular carcinoma, 34 - recurrent hepatocellular carcinoma, and 10 - metastatic liver cancers) in 45 patients were treated with MWA ablation using MR guidance and monitoring. The mean diameter of the liver lesions was 1.7 ± 0.9 cm (range, 0.5-4.6 cm). The 56 liver lesions were divided into 3 groups according to diameter: the <1.0 cm group (17 lesions), the 1.0-2.0 cm group (19 lesions), and the >2.0 cm group (20 lesions). Technical success, technical effectiveness, local tumor progression, procedure duration, and complications were assessed. Primary technical effectiveness was assessed 3 months after the MWA, while local tumor progression was assessed more than 3 months after the MWA. The follow-up time for assessment of treatment response ranged from 12 to 30 months (median, 23 months). RESULTS: The technical success rate was 100%. Primary technical effectiveness was achieved in 52/56 (92.8%) lesions. Local tumor progression was detected in three tumors after initial technical effectiveness. The median duration of the intervention per tumor was 66 min (range, 40-156 min). There were no significant differences between lesion groups in the technical success rate, primary technical effectiveness rate, or local tumor progression rate. There were no major complications following the ablation therapy. CONCLUSIONS: 1.0T open multiparametric MR-guided and MR-monitored MWA for liver cancer is safe and feasible and decreases the risk of local tumor progression; it also provides good primary technique effectiveness rates and is especially suitable when ultrasound and CT facilitated treatments are inappropriate.