Personalized circulating tumor DNA monitoring improves recurrence surveillance and management after curative resection of colorectal liver metastases: a prospective cohort study.

BACKGROUND: Approximately 60% of patients with colorectal liver metastases (CRLM) experience relapse within 2 years after radical resection, previous studies have proven that repeat local treatment (LT) could prolong survival, however, it is difficult to seize the window for LT due to the lack of a high-sensitive surveillance method. In this study, the authors aim to examine the value of longitudinal circulating tumor DNA (ctDNA) in guiding adjuvant chemotherapy, optimizing clinical surveillance strategy, and thereby improving CRLM outcomes. MATERIALS AND METHODS: The authors conducted a prospective clinical trial using a personalized, tumor-informed ctDNA assay to monitor 60 CRLM patients undergoing resection with curative intent. Formalin-fixed paraffin-embedded tumor samples were collected after surgery. Blood samples were collected before surgery, 30 days after surgery (post-OP), and every third month until relapse or up to 2 years. RESULTS: A total of 394 plasma samples from 60 eligible patients were analyzed, with a median follow-up time of 31.3 months. Landmark analyses revealed that detectable ctDNA at post-OP (HR, 4.8), postadjuvant chemotherapy (HR, 6.0), and end-of-treatment (HR, 5.6) were associated with higher recurrence risk ( P <0.001). Post-OP ctDNA positivity served as the only independent prognostic marker in the multivariant analysis (HR, 5.1; P <0.001). Longitudinal ctDNA analysis identified relapsed patients at both sensitivity and specificity of 100%. Most (75%) patients were found with radiological relapse within 6 months after the first detectable ctDNA with a median lead time of 3.5 months. In relapsed patients, 73.2% had oligometastatic disease and 61% were liver-restricted, of which 72.0% received repeat LTs, and 60.0% achieved a secondary no evidence of disease status. CONCLUSIONS: Longitudinal ctDNA monitoring assists in early prediction of relapse, and thereby improves survival of CRLM patients by increased secondary resection rate and secondary no evidence of disease rate.

SNHG17 alters anaerobic glycolysis by resetting phosphorylation modification of PGK1 to foster pro-tumor macrophage formation in pancreatic ductal adenocarcinoma.

BACKGROUND: Within the tumor immune microenvironment (TME), tumor-associated macrophages (TAMs) are crucial in modulating polarization states to influence cancer development through metabolic reprogramming. While long non-coding RNAs (lncRNAs) have been shown to play a pivotal role in the progression of various cancers, the underlying mechanisms by which lncRNAs alter M2 polarization through macrophage metabolism remodeling remain unelucidated. METHODS: RNA sequencing was used to screen for differentially expressed lncRNAs in TAMs and normal tissue-resident macrophages (NTRMs) isolated from pancreatic ductal adenocarcinoma (PDAC) tissues, whilst RT-qPCR and FISH were employed to detect the expression level of SNHG17. Moreover, a series of in vivo and in vitro experiments were conducted to assess the functions of SNHG17 from TAMs in the polarization and glycolysis of M2-like macrophages and in the proliferation and metastasis of pancreatic cancer cells (PCs). Furthermore, Western blotting, RNA pull-down, mass spectrometry, RIP, and dual-luciferase assays were utilized to explore the underlying mechanism through which SNHG17 induces pro-tumor macrophage formation. RESULTS: SNHG17 was substantially enriched in TAMs and was positively correlated with a worse prognosis in PDAC. Meanwhile, functional assays determined that SNHG17 promoted the malignant progression of PCs by enhancing M2 macrophage polarization and anaerobic glycolysis. Mechanistically, SNHG17 could sponge miR-628-5p to release PGK1 mRNA and concurrently interact with the PGK1 protein, activating the pro-tumorigenic function of PGK1 by enhancing phosphorylation at the T168A site of PGK1 through ERK1/2 recruitment. Lastly, SNHG17 knockdown could reverse the polarization status of macrophages in PDAC. CONCLUSIONS: The present study illustrated the essential role of SNHG17 and its molecular mechanism in TAMs derived from PDAC, indicating that SNHG17 might be a viable target for PDAC immunotherapy.

Patient-derived organoids as a platform for drug screening in metastatic colorectal cancer.

Introduction: Most advanced colorectal cancers are aggressive, and there is a lack of effective methods for selecting appropriate anticancer regimens. Patient-derived organoids (PDOs) have emerged as preclinical platforms for modeling clinical responses to cancer therapy. Methods: In this study, we successfully constructed a living biobank with 42 organoids derived from primary and metastatic lesions of metastatic colorectal cancer patients. Tumor tissue was obtained from patients undergoing surgical resection of the primary or metastatic lesion and then used to establish PDOs. Immunohistochemistry (IHC) and drug sensitivity assays were performed to analyze the properties of these organoids. Results: The mCRC organoids were successfully established with an 80% success rate. The PDOs maintained the genetic and phenotypic heterogeneity of their parental tumors. The IC50 values of5-fluorouracil (5-FU), oxaliplatin, and irinotecan (CPT11) were determined for mCRC organoids using drug sensitivity assays. The in vitro chemosensitivity data revealed the potential value of PDOs for clinical applications in predicting chemotherapy response and clinical outcomes in mCRC patients. Discussion: In summary, the PDO model is an effective platform for in vitro assessment of patient-specific drug sensitivity, which can guide personalized treatment decisions for patients with end-stage CRC.

Patient-Derived Organoids from Colorectal Cancer with Paired Liver Metastasis Reveal Tumor Heterogeneity and Predict Response to Chemotherapy.

There is no effective method to predict chemotherapy response and postoperative prognosis of colorectal cancer liver metastasis (CRLM) patients. Patient-derived organoid (PDO) has become an important preclinical model. Herein, a living biobank with 50 CRLM organoids derived from primary tumors and paired liver metastatic lesions is successfully constructed. CRLM PDOs from the multiomics levels (histopathology, genome, transcriptome and single-cell sequencing) are comprehensively analyzed and confirmed that this organoid platform for CRLM could capture intra- and interpatient heterogeneity. The chemosensitivity data in vitro reveal the potential value of clinical application for PDOs to predict chemotherapy response (FOLFOX or FOLFIRI) and clinical prognosis of CRLM patients. Taken together, CRLM PDOs can be utilized to deliver a potential application for personalized medicine.

Assay establishment and validation of a high-throughput organoid-based drug screening platform.

BACKGROUND: Organoids are three-dimensional structures that closely recapitulate tissue architecture and cellular composition, thereby holding great promise for organoid-based drug screening. Although growing in three-dimensional provides the possibility for organoids to recapitulate main features of corresponding tissues, it makes it incommodious for imaging organoids in two-dimensional and identifying surviving organoids from surrounding dead cells after organoids being treated by irradiation or chemotherapy. Therefore, significant work remains to establish high-quality controls to standardize organoid analyses and make organoid models more reproducible. METHODS: In this study, the Z-stack imaging technique was used for the imaging of three-dimensional organoids to gather all the organoids' maximum cross sections in one imaging. The combination of live cell staining fluorescent dye Calcein-AM and ImageJ assessment was used to analyze the survival of organoids treated by irradiation or chemotherapy. RESULTS: We have established a novel quantitative high-throughput imaging assay that harnesses the scalability of organoid cultures. Using this assay, we can capture organoid growth over time, measure multiple whole-well organoid readouts, and show the different responses to drug treatments. CONCLUSIONS: In summary, combining the Z-stack imaging technique and fluorescent labeling methods, we established an assay for the imaging and analysis of three-dimensional organoids. Our data demonstrated the feasibility of using organoid-based platforms for high-throughput drug screening assays.

LncRNA-PACERR induces pro-tumour macrophages via interacting with miR-671-3p and m6A-reader IGF2BP2 in pancreatic ductal adenocarcinoma.

BACKGROUND: LncRNA-PACERR plays critical role in the polarization of tissue-associated macrophages (TAMs). In this study, we found the function and molecular mechanism of PACERR in TAMs to regulate pancreatic ductal adenocarcinoma (PDAC) progression. METHODS: We used qPCR to analyse the expression of PACERR in TAMs and M1-tissue-resident macrophages (M1-NTRMs) which were isolated from 46 PDAC tissues. The function of PACERR on macrophages polarization and PDAC proliferation, migration and invasion were confirmed through in vivo and in vitro assays. The molecular mechanism of PACERR was discussed via fluorescence in situ hybridization (FISH), RNA pull-down, ChIP-qPCR, RIP-qPCR and luciferase assays. RESULTS: LncRNA-PACERR was high expression in TAMs and associated with poor prognosis in PDAC patients. Our finding validated that LncRNA-PACERR increased the number of M2-polarized cells and facilized cell proliferation, invasion and migration in vitro and in vivo. Mechanistically, LncRNA-PACERR activate KLF12/p-AKT/c-myc pathway by binding to miR-671-3p. And LncRNA-PACERR which bound to IGF2BP2 acts as an m6A-dependent manner to enhance the stability of KLF12 and c-myc in cytoplasm. In addition, the promoter of LncRNA-PACERR was a target of KLF12 and LncRNA-PACERR recruited EP300 to increase the acetylation of histone by interacting with KLF12 in nucleus. CONCLUSIONS: This study found that LncRNA-PACERR functions as key regulator of TAMs in PDAC microenvironment and revealed the novel mechanisms in cytoplasm and in nucleus.

Treatment of hyperlipidemic acute pancreatitis with modified Dachengqi decoction combining with conventional therapy based on "six-hollow-organs to be unblocked" theory.

BACKGROUND: The increase of triglyceride (TG) can induce coronary heart disease, atherosclerosis, pancreatitis and other diseases and is the most common inducing factor of acute pancreatitis (AP) second only to biliary tract disease and drinking. The pathogenesis of hyperlipidemic acute pancreatitis (HLAP) is not exactly clear, but it may be related to the toxic effect of the increase of free fatty acids produced by TG decomposition on the pancreas itself, microcirculation disorder of the pancreas, and calcium overload. At present, non-surgical therapy is the main treatment for HLAP. The key to preventing recurrence is to reduce blood lipids, change the diet structure, and reduce weight. This study aimed to treat HLAP with modified Dachengqi decoction combined with conventional therapy, based on the "six-hollow-organs to be unblocked" theory. METHODS: Forty patients with HLAP who received treatment in the First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine and Puding County Hospital of Traditional Chinese Medicine from September 2016 to August 2019 were selected and divided into a control group and an intervention group, each with 20 cases, following a random number table. The control group was treated with conventional therapy while the intervention group was treated with modified Dachengqi decoction combined with conventional therapy. RESULTS: After treatment, the cure rate and the total effective rate were 60% and 95% respectively in the intervention group, and 25% and 75% respectively in the control group (P<0.05). The TG, serum amylase, leukocyte count, and neutrophil ratio of the two groups decreased significantly after treatment, and there was a greater decrease in the intervention group than that in the control group, with this being significantly different between the two groups. The gastrointestinal function score, total score of the acute physiology and chronic health evaluation II (APACHE II), and the pain score of the visual analog scale (VAS) decreased markedly in the two groups after the treatment, with scores in the intervention group being significantly lower than those in the control group (P<0.05). CONCLUSIONS: Modified Dachengqi decoction combined with conventional therapy has a better therapeutic effect on HLAP than conventional therapy.

Narrative review of the mechanisms of action of dachengqi decoction in the treatment of hyperlipidemic pancreatitis on six-hollow-organs to be unblocked theory.

The incidence rate of acute pancreatitis (AP) caused by hyperlipidemia is increasing year by year. The primary treatment goal is to reduce blood lipids rapidly. On the theory of "Six-hollow-organs to be unblocked" we used dachengqi decoction (original prescription of Zhang Zhongjing in Shanghan Lun) to block the peroxisome proliferator-activated receptor γ (PPARG) pathway and rapidly reduce blood lipid to achieve the purpose of treating hyperlipidemic acute pancreatitis (HLAP). In this review, we summarize the etiology and pathogenesis of HLAP and the progress of traditional Chinese medicine in treating HLAP. The mechanisms of action of dachengqi decoction in the treatment of HLAP and the involvement of the PPARG pathway were discussed. In brief, the dachengqi decoction has the effect of resolving phlegm and clearing waste substances and can improve intestinal function; can inhibits the production of interleukin-1, interleukin-6, and tumor necrosis factor-α, and reduces the damage of SIRS to human body; also it improves the microcirculation system by inhibiting the production of inflammatory factors, reducing, or eliminating the damage to vascular endothelial cells and microvessels, and improving vascular permeability. The clarification of the mechanisms of action of the drug is conducive to the extensive clinical application of the classical formula.

Ruxolitinib protects lipopolysaccharide (LPS)-induced sepsis through inhibition of nitric oxide production in mice.

BACKGROUND: Ruxolitinib is an inhibitor of Janus kinases (JAK) 1/2. It was authorised recently by the U.S. Food and Drug Administration (FDA) as a new Myelofibrosis treatment. In this study, we identified ruxolitinib as a new inhibitor of nitric oxide (NO) production in response to lipopolysaccharide (LPS)stimulation of RAW 264.7 cells. METHODS: In vitro direct effects of ruxolitinib were determined through NO production on RAW 264.7 cells. Also the expression level of iNOS, TNF-α and IL-6 were detected by Western Blotting and qRT-PCR. In vivo therapeutic effects of ruxolitinib on sepsis were evaluated by an endotoxemia model with C57 mice. The survival was calculated and histopathological damage of organs was observed by HE. Cytokines in serum were detected by Mouse Cytokine Array Panel. RESULTS: Ruxolitinib was found to significantly reduce NO production, inducible nitric oxide synthase (iNOS), TNF-α, and IL-6 expression, suggesting that ruxolitinib blocks LPS signaling that leads to pro-inflammatory factor expression. Furthermore, the inhibitory effects of ruxolitinib contributed to the survival of septic mice by 70% and pro-inflammatory cytokines in serum declined apparently. The results taken together indicate that ruxolitinib can significantly suppress LPS-stimulated NO production and improve the survival of septic mice, perhaps by interfering with the NF-κB pathway. CONCLUSIONS: These findings suggest ruxolitinib might be a possible therapeutic candidate for sepsis therapy.

The Impact of Blood Type O on Major Outcomes in Patients With Severe Burns.

ABO blood type has been reported to be a predictor of poor prognosis in critically ill patients. Here, we aim to correlate different blood types with clinical outcomes in patients with severe burns. We conducted a single-center retrospective cohort study by enrolling patients with severe burn injuries (≥40% TBSA) between January 2012 and December 2017. Baseline characteristics and clinical outcomes were compared between disparate ABO blood types (type O vs non-O type). Multivariate logistic and linear regression analyses were performed to identify an association between ABO blood type and clinical outcomes, including in-hospital mortality, the development of acute kidney injury (AKI), and hospital or ICU length of stay. A total of 141 patients were finally enrolled in the current study. Mortality was significantly higher in patients with type O blood compared with those of other blood types. The development of AKI was significantly higher in patients with blood type O vs non-O blood type (P = .001). Multivariate analysis demonstrated that blood type O was independently associated with in-hospital mortality and AKI occurrence after adjusting for other potential confounders. Our findings indicated the blood type O was an independent risk factor of both increased mortality and the development of AKI postburn. More prudent and specific treatments are required in treating these patients to avoid poor prognosis.

Li15P4S16Cl3, a Lithium Chlorothiophosphate as a Solid-State Ionic Conductor.

Tremendous efforts have been devoted to the design of solid Li+ electrolytes and the development of all-solid-state batteries. Compared with conventional Li-ion batteries, which use flammable liquid organic electrolytes, all-solid-state batteries show significant advantages in safety. In this work, a novel lithium chlorothiophosphate compound, Li15P4S16Cl3, is discovered. The crystal structure and electrochemical properties are investigated. Li15P4S16Cl3 can be synthesized as a pure phase via a facile solid-state reaction by heating a ball-milled mixture of Li2S, P2S5, and LiCl at 360 °C. The crystal structure of Li15P4S16Cl3 was refined against neutron and synchrotron powder X-ray diffraction data, revealing that it crystallizes in the space group I4̅3d. The Li+ transport in Li15P4S16Cl3 was also investigated by multiple solid-state NMR methods, including variable-temperature NMR line-shape analysis, NMR relaxometry, and pulsed-field-gradient NMR. Li15P4S16Cl3 shows good thermodynamic stability and can be synthesized at relatively low temperature. Although it exhibits a low ionic conductivity at room temperature, it can serve as a new motif crystal structure for the design and development of new solid-state electrolytes.

Unsupervised discovery of solid-state lithium ion conductors.

Although machine learning has gained great interest in the discovery of functional materials, the advancement of reliable models is impeded by the scarcity of available materials property data. Here we propose and demonstrate a distinctive approach for materials discovery using unsupervised learning, which does not require labeled data and thus alleviates the data scarcity challenge. Using solid-state Li-ion conductors as a model problem, unsupervised materials discovery utilizes a limited quantity of conductivity data to prioritize a candidate list from a wide range of Li-containing materials for further accurate screening. Our unsupervised learning scheme discovers 16 new fast Li-conductors with conductivities of 10-4-10-1 S cm-1 predicted in ab initio molecular dynamics simulations. These compounds have structures and chemistries distinct to known systems, demonstrating the capability of unsupervised learning for discovering materials over a wide materials space with limited property data.

Strategies Based on Nitride Materials Chemistry to Stabilize Li Metal Anode.

Lithium metal battery is a promising candidate for high-energy-density energy storage. Unfortunately, the strongly reducing nature of lithium metal has been an outstanding challenge causing poor stability and low coulombic efficiency in lithium batteries. For decades, there are significant research efforts to stabilize lithium metal anode. However, such efforts are greatly impeded by the lack of knowledge about lithium-stable materials chemistry. So far, only a few materials are known to be stable against Li metal. To resolve this outstanding challenge, lithium-stable materials have been uncovered out of chemistry across the periodic table using first-principles calculations based on large materials database. It is found that most oxides, sulfides, and halides, commonly studied as protection materials, are reduced by lithium metal due to the reduction of metal cations. It is discovered that nitride anion chemistry exhibits unique stability against Li metal, which is either thermodynamically intrinsic or a result of stable passivation. The results here establish essential guidelines for selecting, designing, and discovering materials for lithium metal protection, and propose multiple novel strategies of using nitride materials and high nitrogen doping to form stable solid-electrolyte-interphase for lithium metal anode, paving the way for high-energy rechargeable lithium batteries.

Origin of fast ion diffusion in super-ionic conductors.

Super-ionic conductor materials have great potential to enable novel technologies in energy storage and conversion. However, it is not yet understood why only a few materials can deliver exceptionally higher ionic conductivity than typical solids or how one can design fast ion conductors following simple principles. Using ab initio modelling, here we show that fast diffusion in super-ionic conductors does not occur through isolated ion hopping as is typical in solids, but instead proceeds through concerted migrations of multiple ions with low energy barriers. Furthermore, we elucidate that the low energy barriers of the concerted ionic diffusion are a result of unique mobile ion configurations and strong mobile ion interactions in super-ionic conductors. Our results provide a general framework and universal strategy to design solid materials with fast ionic diffusion.

Negating interfacial impedance in garnet-based solid-state Li metal batteries.

Garnet-type solid-state electrolytes have attracted extensive attention due to their high ionic conductivity, approaching 1 mS cm-1, excellent environmental stability, and wide electrochemical stability window, from lithium metal to ∼6 V. However, to date, there has been little success in the development of high-performance solid-state batteries using these exceptional materials, the major challenge being the high solid-solid interfacial impedance between the garnet electrolyte and electrode materials. In this work, we effectively address the large interfacial impedance between a lithium metal anode and the garnet electrolyte using ultrathin aluminium oxide (Al2O3) by atomic layer deposition. Li7La2.75Ca0.25Zr1.75Nb0.25O12 (LLCZN) is the garnet composition of choice in this work due to its reduced sintering temperature and increased lithium ion conductivity. A significant decrease of interfacial impedance, from 1,710 Ω cm2 to 1 Ω cm2, was observed at room temperature, effectively negating the lithium metal/garnet interfacial impedance. Experimental and computational results reveal that the oxide coating enables wetting of metallic lithium in contact with the garnet electrolyte surface and the lithiated-alumina interface allows effective lithium ion transport between the lithium metal anode and garnet electrolyte. We also demonstrate a working cell with a lithium metal anode, garnet electrolyte and a high-voltage cathode by applying the newly developed interface chemistry.

Origin of Outstanding Stability in the Lithium Solid Electrolyte Materials: Insights from Thermodynamic Analyses Based on First-Principles Calculations.

First-principles calculations were performed to investigate the electrochemical stability of lithium solid electrolyte materials in all-solid-state Li-ion batteries. The common solid electrolytes were found to have a limited electrochemical window. Our results suggest that the outstanding stability of the solid electrolyte materials is not thermodynamically intrinsic but is originated from kinetic stabilizations. The sluggish kinetics of the decomposition reactions cause a high overpotential leading to a nominally wide electrochemical window observed in many experiments. The decomposition products, similar to the solid-electrolyte-interphases, mitigate the extreme chemical potential from the electrodes and protect the solid electrolyte from further decompositions. With the aid of the first-principles calculations, we revealed the passivation mechanism of these decomposition interphases and quantified the extensions of the electrochemical window from the interphases. We also found that the artificial coating layers applied at the solid electrolyte and electrode interfaces have a similar effect of passivating the solid electrolyte. Our newly gained understanding provided general principles for developing solid electrolyte materials with enhanced stability and for engineering interfaces in all-solid-state Li-ion batteries.

Accelerated materials design of Na0.5Bi0.5TiO3 oxygen ionic conductors based on first principles calculations.

We perform a first principles computational study of designing the Na0.5Bi0.5TiO3 (NBT) perovskite material to increase its oxygen ionic conductivity. In agreement with the previous experiments, our computation results confirm fast oxygen ionic diffusion and good stability of the NBT material. The oxygen diffusion mechanisms in this new material were systematically investigated, and the effects of local atomistic configurations and dopants on oxygen diffusion were revealed. Novel doping strategies focusing on the Na/Bi sublattice were predicted and demonstrated by the first principles calculations. In particular, the K doped NBT compound achieved good phase stability and an order of magnitude increase in oxygen ionic conductivity of up to 0.1 S cm(-1) at 900 K compared to the previous Mg doped compositions. This study demonstrated the advantages of first principles calculations in understanding the fundamental structure-property relationship and in accelerating the materials design of the ionic conductor materials.

Nanoparticle-based adjuvant for enhanced protective efficacy of DNA vaccine Ag85A-ESAT-6-IL-21 against Mycobacterium tuberculosis infection.

The goal of this study was to evaluate the protective efficacy of a cationic nanoparticle-based DNA vaccine expressing antigen 85A (Ag85A) and 6-kDa early secretory antigen target (ESAT-6) of Mycobacterium tuberculosis as well as cytokine interleukin-21 (IL-21) against M. tuberculosis infection. The results of this indicated that the anti-M. tuberculosis immune responses were induced in mice that had received the different DNA vaccines. More importantly, compared with using DNA vaccine Ag85A-ESAT-6-IL-21 alone, the nanoparticle-based DNA vaccine Ag85A-ESAT-6-IL-21 showed a statistically significant increase in the protective efficacy against M. tuberculosis infection in the immunized mice. We concluded that the nanoparticle-based DNA vaccine induced a strong immune response and markedly inhibited the growth of the M. tuberculosis in the mice. These findings highlighted the potential utility of Fe3O4-Glu-polyethyleneimine nanoparticles encapsulated with the DNA vaccine as a prophylactic vaccine in the M. tuberculosis-infected mouse model. FROM THE CLINICAL EDITOR: This study emphasizes the potential utility of Fe3O4-Glu-polyethyleneimine nanoparticles encapsulated with DNA vaccine against TB as a prophylactic vaccine. The authors demonstrated a strong immune response and marked growth inhibition of mycobacterium tuberculosis in the mice.