Targeted delivery of FAK siRNA by engineered exosomes to reverse cetuximab resistance via activating paraptosis in colon cancer.

BACKGROUND: Cetuximab is extensively used in the treatment of metastatic colorectal cancer (mCRC). However, resistance poses a significant challenge to successful therapy. Recently, paraptosis, a non-classical programmed cell death, has garnered increased attention for its potential application value in antitumor treatments. We aimed to identify the essential pathways and signaling molecules involved in paraptosis inhibition and select them as therapeutic targets in cetuximab resistance. Additionally, engineered exosome technology is used as a drug delivery system with both targeted and effector properties. RESULTS: By comparing the differential expression of paraptosis-related genes between drug-resistant colon cancer cells and sensitive cells, it was observed that the paraptosis level induced by cetuximab was significantly downregulated in drug-resistant cells. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis identified the focal adhesion kinase (FAK) signaling pathway as a key pathway involved in the suppression of paraptosis. The biological function of FAK in cetuximab-resistant cells was investigated through cell morphology observation, CCK-8 assay, colony formation assay, RT-qPCR, Western Blot, and loss-of-function experiments. The results showed that the FAK signaling pathway was significantly upregulated in cetuximab-resistant colon cancer cells, and siRNA interference targeting FAK could notably inhibit cell proliferation while upregulating the paraptosis level. Based on this, engineered colon cancer cells targeted and FAK siRNA loaded exosomes (CT-Exo-siFAK1) were constructed. In vitro experiments, CT-Exo-siFAK1 could effectively activate paraptosis and inhibit the proliferation of drug-resistant colon cancer cells. In vivo experiments also confirmed that CT-Exo-siFAK1 significantly suppressed tumor growth and metastasis while upregulating the paraptosis level. CONCLUSION: This study suggests that FAK signaling pathway-mediated inhibition of paraptosis levels is crucial in the sensitivity of cetuximab targeted therapy in colon cancer, and the use of engineered exosomes to deliver FAK siRNA may be an effective strategy to reverse cetuximab resistance.

Multi-nuclear sodium, diffusion, and perfusion MRI in human gliomas.

PURPOSE: There is limited knowledge about the associations between sodium and proton MRI measurements in brain tumors. The purpose of this study was to quantify intra- and intertumoral correlations between sodium, diffusion, and perfusion MRI in human gliomas. METHODS: Twenty glioma patients were prospectively studied on a 3T MRI system with multinuclear capabilities. Three mutually exclusive tumor volumes of interest (VOIs) were segmented: contrast-enhancing tumor (CET), T2/FLAIR hyperintense non-enhancing tumor (NET), and necrosis. Median and voxel-wise associations between apparent diffusion coefficient (ADC), normalized relative cerebral blood volume (nrCBV), and normalized sodium measurements were quantified for each VOI. RESULTS: Both relative sodium concentration and ADC were significantly higher in areas of necrosis compared to NET (P = 0.003 and P = 0.008, respectively) and CET (P = 0.02 and P = 0.02). Sodium concentration was higher in CET compared to NET (P = 0.04). Sodium and ADC were higher in treated compared to treatment-naïve gliomas within NET (P = 0.006 and P = 0.01, respectively), and ADC was elevated in CET (P = 0.03). Median ADC and sodium concentration were positively correlated across patients in NET (r = 0.77, P < 0.0001) and CET (r = 0.84, P < 0.0001), but not in areas of necrosis (r = 0.45, P = 0.12). Median nrCBV and sodium concentration were negatively correlated across patients in areas of NET (r=-0.63, P = 0.003). Similar associations were observed when examining voxel-wise correlations within VOIs. CONCLUSION: Sodium MRI is positively correlated with proton diffusion MRI measurements in gliomas, likely reflecting extracellular water. Unique areas of multinuclear MRI contrast may be useful in future studies to understand the chemistry of the tumor microenvironment.

Mechanotransduction-induced glycolysis epigenetically regulates a CXCL1-dominant angiocrine signaling program in liver sinusoidal endothelial cells in vitro and in vivo.

BACKGROUND & AIMS: Liver sinusoidal endothelial cells (LSECs) are ideally situated to sense stiffness and generate angiocrine programs that potentially regulate liver fibrosis and portal hypertension. We explored how specific focal adhesion (FA) proteins parlay LSEC mechanotransduction into stiffness-induced angiocrine signaling in vitro and in vivo. METHODS: Primary human and murine LSECs were placed on gels with incremental stiffness (0.2 kPa vs. 32 kPa). Cell response was studied by FA isolation, actin polymerization assay, RNA-sequencing and electron microscopy. Glycolysis was assessed using radioactive tracers. Epigenetic regulation of stiffness-induced genes was analyzed by chromatin-immunoprecipitation (ChIP) analysis of histone activation marks, ChIP sequencing and circularized chromosome conformation capture (4C). Mice with LSEC-selective deletion of glycolytic enzymes (Hk2fl/fl/Cdh5cre-ERT2) or treatment with the glycolysis inhibitor 3PO were studied in portal hypertension (partial ligation of the inferior vena cava, pIVCL) and early liver fibrosis (CCl4) models. RESULTS: Glycolytic enzymes, particularly phosphofructokinase 1 isoform P (PFKP), are enriched in isolated FAs from LSECs on gels with incremental stiffness. Stiffness resulted in PFKP recruitment to FAs, which paralleled an increase in glycolysis. Glycolysis was associated with expansion of actin dynamics and was attenuated by inhibition of integrin ß1. Inhibition of glycolysis attenuated a stiffness-induced CXCL1-dominant angiocrine program. Mechanistically, glycolysis promoted CXCL1 expression through nuclear pore changes and increases in NF-kB translocation. Biochemically, this CXCL1 expression was mediated through spatial re-organization of nuclear chromatin resulting in formation of super-enhancers, histone acetylation and NF-kB interaction with the CXCL1 promoter. Hk2fl/fl/Cdh5cre-ERT2 mice showed attenuated neutrophil infiltration and portal hypertension after pIVCL. 3PO treatment attenuated liver fibrosis in a CCl4 model. CONCLUSION: Glycolytic enzymes are involved in stiffness-induced angiocrine signaling in LSECs and represent druggable targets in early liver disease. LAY SUMMARY: Treatment options for liver fibrosis and portal hypertension still represent an unmet need. Herein, we uncovered a novel role for glycolytic enzymes in promoting stiffness-induced angiocrine signaling, which resulted in inflammation, fibrosis and portal hypertension. This work has revealed new targets that could be used in the prevention and treatment of liver fibrosis and portal hypertension.

Coordinated signaling of activating transcription factor 6α and inositol-requiring enzyme 1α regulates hepatic stellate cell-mediated fibrogenesis in mice.

Liver injury and the unfolded protein response (UPR) are tightly linked, but their relationship differs with cell type and injurious stimuli. UPR initiation promotes hepatic stellate cell (HSC) activation and fibrogenesis, but the underlying mechanisms are unclear. Despite the complexity and overlap downstream of UPR transducers inositol-requiring protein 1α (IRE1α), activating transcription factor 6α (ATF6α), and protein kinase RNA-like ER kinase (PERK), previous research in HSCs primarily focused on IRE1α. Here, we investigated the fibrogenic role of ATF6α or PERK in vitro and HSC-specific UPR signaling in vivo. Overexpression of ATF6α, but not the PERK effector activating transcription factor 4 (ATF4), promoted HSC activation and fibrogenic gene transcription in immortalized HSCs. Furthermore, ATF6α inhibition through Ceapin-A7, or Atf6a deletion, disrupted transforming growth factor ß (TGFß)-mediated activation of primary human hepatic stellate cells (hHSCs) or murine hepatic stellate cells (mHSCs), respectively. We investigated the fibrogenic role of ATF6α in vivo through conditional HSC-specific Atf6a deletion. Atf6aHSCΔ/Δ mice displayed reduced fibrosis and HSC activation following bile duct ligation (BDL) or carbon tetrachloride (CCl4)-induced injury. The Atf6aHSCΔ/Δ phenotype differed from HSC-specific Ire1a deletion, as Ire1aHSCΔ/Δ mice showed reduced fibrogenic gene transcription but no changes in fibrosis compared with Ire1afl/fl mice following BDL. Interestingly, ATF6α signaling increased in Ire1aΔ/Δ HSCs, whereas IRE1α signaling was upregulated in Atf6aΔ/Δ HSCs. Finally, we asked whether co-deletion of Atf6a and Ire1a additively limits fibrosis. Unexpectedly, fibrosis worsened in Atf6aHSCΔ/ΔIre1aHSCΔ/Δ mice following BDL, and Atf6aΔ/ΔIre1aΔ/Δ mHSCs showed increased fibrogenic gene transcription. ATF6α and IRE1α individually promote fibrogenic transcription in HSCs, and ATF6α drives fibrogenesis in vivo. Unexpectedly, disruption of both pathways sensitizes the liver to fibrogenesis, suggesting that fine-tuned UPR signaling is critical for regulating HSC activation and fibrogenesis.NEW & NOTEWORTHY ATF6α is a critical driver of hepatic stellate cell (HSC) activation in vitro. HSC-specific deletion of Atf6a limits fibrogenesis in vivo despite increased IRE1α signaling. Conditional deletion of Ire1α from HSCs limits fibrogenic gene transcription without impacting overall fibrosis. This could be due in part to observed upregulation of the ATF6α pathway. Dual loss of Atf6a and Ire1a from HSCs worsens fibrosis in vivo through enhanced HSC activation.

Intra-abdominal hemorrhage following 739 consecutive pancreaticoduodenectomy: Risk factors and treatments.

BACKGROUND AND AIM: Post-pancreaticoduodenectomy hemorrhage (PPH) is a potentially lethal complication. The objective of this study was to explore the risk factors of PPH and to evaluate the treatment options. METHODS: Clinical data of 739 consecutive patients undergoing pancreaticoduodenectomy between 2009 and 2017 were collected from a prospectively maintained database. Univariate and multivariate analysis was performed by logistic regression model to evaluate potential risk factors associated with early and late PPH. RESULTS: The morbidity of PPH was 8.7% (64/739), while the mortality was 12.5% (8/64). Twenty-two (34.4%) patients developed PPH within postoperative day 1 (early PPH) whereas 42 (65.6%) patients after postoperative day 1 (late PPH). No significant risk factor was identified associated with early PPH, whereas pancreatic duct diameter < 0.4 cm, and intra-abdominal complications, such as pancreatic fistula, intra-abdominal abscess, and delayed gastric emptying, were independently correlated with late PPH. There were 10 (15.6%) grade A, 28 (43.8%) grade B, and 26 (40.6%) grade C bleedings. The bleeding sites were verified by endoscopy, angiography, and/or exploratory laparotomy in 23 of 54 (42.6%) patients with grade B or C hemorrhage. Seven out of nine (78%) patients with arterial bleeding were cured by angiography and embolization, while 10 of 11 (90.9%) patients with anastomotic, venous, or retroperitoneum bleeding were rescued by laparotomy. Ten patients with grade A and 22 patients with grade B or C hemorrhage were treated successfully by blood transfusion and hemostatic medications. CONCLUSIONS: Hemorrhage following pancreaticoduodenectomy is a common and lethal complication. Treatment strategies should be tailored according to different etiologies.

Novel laparoscopic training system with continuously perfused ex-vivo porcine liver for hepatobiliary surgery.

OBJECTIVE: To introduce a novel laparoscopic training system with a continuously perfused ex-vivo porcine liver for hepatobiliary surgery. BACKGROUND: Existing models for laparoscopic training, such as box trainers and virtual reality simulators, often fail to provide holistic training and real haptic feedback. We have formulated a new training system that addresses these problems. METHODS: Real-Liver Laptrainer consists of a porcine liver, customized mannequin, ex-vivo machine perfusion system, and monitoring software. We made a detailed comparison of Real-Liver Laptrainer with the LapSim virtual reality simulator and the FLS Trainer Box systems. Five laparoscopic surgeons assessed the new system on multiple features. We assessed the performances of 43 trainees who used the new system to perform laparoscopic cholecystectomy (LC) three times. RESULTS: Real-Liver Laptrainer offered more functions and better tactile feedback than the FLS or LapSim system. All five surgeons graded the quality of the new system as realistic. The utility of the system for training was scored as 3.6 ± 1.1 on a scale of 1-5. Between the first and third attempts, the number of successfully performed LCs increased (9 vs 14 vs 23; P = .011), while the numbers of liver damage incidents (25 vs. 21 vs. 18, P = .303) and gallbladder perforations decreased (17 vs. 12 vs. 9, P = .163). The mean LC operation time significantly decreased (63 vs. 50 vs. 44, P < .0001). CONCLUSION: Real-Liver Laptrainer is a feasible, stable, and practical training model that has potential for improving the laparoscopic skills of surgeons.

An optimized procedure for stained bloodless anatomic hepatectomy in canines.

BACKGROUND: Poloxamer 407 (P407) is a thermosensitive polymer that can gelatinize at body temperature and dissolve below critical temperature. The aim of this study was to evaluate an optimized procedure for hepatectomy, in which the target liver section was stained with methylene blue, and the blood inflow was occluded with P407. METHODS: Twelve dogs were randomized into two equal groups. The conventional group (CG) underwent unstained liver resection with the hemi-Pringle maneuver for blood control. After angiography, the optimized group (OG) was cannulated to the target lobar hepatic artery via the femoral artery and to the target segmental portal vein via a branch of the splenic vein. The artery was then occluded with P407, whereas the vein was administered methylene blue and P407 sequentially before excision along the stained border. Blood specimens and necropsy were acquired periodically. RESULTS: The stained resection margins were clearly visualized and were accompanied by negligible blood loss. The occlusion duration was significantly reduced from 24.5 ± 2.3 min in the conventional group to 18.5 ± 4.9 min in the OG (P < 0.05). The aspartate aminotransferase and alanine aminotransferase levels were less elevated in the OG postoperatively. No significant evidence of pathology was detected in either group. CONCLUSIONS: This optimized procedure represents an easy, time-saving and effective approach for stained anatomic hepatectomy with temporary intravascular blood occlusion.

Mass Continuous Suture versus Layered Interrupted Suture in Transverse Abdominal Incision Closure after Liver Resection.

BACKGROUND: Abdominal incision closure technique seriously influences patient prognosis. Most studies have focused on the different suture techniques and materials on midline incision, while little data are available in wide transverse or oblique incisions after liver resection (LR). The aim of the present study is to compare the two major incision suture methods after LR in our institute: Mass continuous suture (group P) and layered interrupted suture (group S). STUDY DESIGN: 258 patients undergoing LR with abdominal transverse or oblique incisions were prospectively enrolled. They were divided into two groups according to different abdominal incision suture methods and compared with the preoperative, intraoperative parameters, and postoperative wound complications. RESULTS: There were 118 patients in group P and 140 patients in group S, which was similar in general condition, primary disease, liver, and renal function. Incision length, total operation time, intraoperative blood loss, or perioperative antibiotics use were not different between the two groups. However, abdominal incision closure time and interval time for stitches removing after operation was significantly shorter in group P than group S (both p < 0.001). After a median follow-up of 16 months, the incidence of wound infection and fat liquefaction was more than two times higher in group S than group P, which, however, was not statistically different. Moreover, there was no difference in wound disruption or incisional hernia between the two groups. CONCLUSIONS: Although similar in occurrence of postoperative wound complications, mass continuous suture with polydioxanone seemed to be more timesaving in incision closure and motivated in wound healing.

Splenorenal shunt via magnetic compression technique: a feasibility study in canine and cadaver.

INTRODUCTION: The concept of magnetic compression technique (MCT) has been accepted by surgeons to solve a variety of surgical problems. In this study, we attempted to explore the feasibility of a splenorenal shunt using MCT in canine and cadaver. MATERIAL AND METHODS: The diameters of the splenic vein (SV), the left renal vein (LRV), and the vertical interval between them, were measured in computer tomography (CT) images obtained from 30 patients with portal hypertension and in 20 adult cadavers. The magnetic devices used for the splenorenal shunt were then manufactured based on the anatomic parameters measured above. The observation of the anatomical structure showed there were no special structural tissues or any important organs between SV and LRV. Then the magnetic compression splenorenal shunt procedure was performed in three dogs and five cadavers. Seven days later, the necrotic tissue between the two magnets was shed and the magnets were removed with the anchor wire. RESULTS: The feasibility of splenorenal shunt via MCT was successfully shown in both canine and cadaver, thus providing a theoretical support for future clinical application.

[Rectovaginal Fistula Stage-one Repair Device Based on Magnetic Compression Technique].

The magnamosis device for stage-one repair of the rectovaginal tistula consists of two arc magnets. Drawing the interrupting thread along the fistula margin via the vaginal side, and pulling the string to arrange the magnets at the fistula base along the long axis of the vagina, we made the magnamosis device automatically clipped to seal the fistula. After removing the threads we kept the device for 2-4 weeks till the natural detachment of it when the compressed tissue in between healed after vascular necrosis. This device utilizing the unique ability of magnamosis to fulfill anastomosis under inflammatory infected state reduces the current high relapse rate and colostomy drawbacks of the conventional rectovaginal neoplasty.

Liver sinusoidal endothelial cells contribute to portal hypertension through collagen type IV-driven sinusoidal remodeling.

Portal hypertension (PHTN) is a severe complication of liver cirrhosis and is associated with intrahepatic sinusoidal remodeling induced by sinusoidal resistance and angiogenesis. Collagen type IV (COL4), a major component of basement membrane, forms in liver sinusoids upon chronic liver injury. However, the role, cellular source, and expression regulation of COL4 in liver diseases are unknown. Here, we examined how COL4 is produced and how it regulates sinusoidal remodeling in fibrosis and PHTN. Human cirrhotic liver sample RNA sequencing showed increased COL4 expression, which was further verified via immunofluorescence staining. Single-cell RNA sequencing identified liver sinusoidal endothelial cells (LSECs) as the predominant source of COL4 upregulation in mouse fibrotic liver. In addition, COL4 was upregulated in a TNF-α/NF-κB-dependent manner through an epigenetic mechanism in LSECs in vitro. Indeed, by utilizing a CRISPRi-dCas9-KRAB epigenome-editing approach, epigenetic repression of the enhancer-promoter interaction showed silencing of COL4 gene expression. LSEC-specific COL4 gene mutation or repression in vivo abrogated sinusoidal resistance and angiogenesis, which thereby alleviated sinusoidal remodeling and PHTN. Our findings reveal that LSECs promote sinusoidal remodeling and PHTN during liver fibrosis through COL4 deposition.

The m6A reader YTHDF1 attenuates fulminant hepatitis via MFG-E8 translation in an m6A dependent manner.

Background and Aims: N6-methyladenosine (m6A) is the most common post-transcriptional modification of RNA in eukaryotes, which has been demonstrated to play important roles in various biological processes. However, its roles in fulminant hepatitis remain largely unknown. In the current study, YTHDF1 expression was found to be significantly downregulated in the livers among patients, as well as murine models with fulminant hepatitis versus normal controls. Thus, we hypothesized that YTHDF1 protects against fulminant hepatitis and investigated the underlying molecular mechanisms. Methods: Fulminant hepatitis was induced by D-GalN/LPS in conventional YTHDF1 knockout (YTHDF1-/-) mice, hepatocyte-specific YTHDF1 overexpression (AAV8- YTHDF1) mice, and corresponding control mice. Primary hepatocytes were cultured and subjected to LPS insult in vitro. Hepatic histology, cell death, oxidative stress and mitochondrial function were examined to assess liver damage. The molecular mechanisms of YTHDF1 function were explored using multi-omics analysis. Results: Ablation of YTHDF1 exacerbated hepatic apoptosis and reactive oxygen species (ROS) production and increased the number of aberrant mitochondria, while YTHDF1 overexpression resulted in the opposite effects. Multiomics analysis identified MFG-E8 as the direct target of YTHDF1. YTHDF1 augmented the translation of MFG-E8 in an m6A-dependent manner without effect on its mRNA expression, thereby restoring mitochondrial function. Additionally, administration of MFG-E8 almost completely reversed the YTHDF1 deficiency-mediated exacerbation of liver injury. Conclusions: The current study suggested that the m6A reader YTHDF1 alleviates cell death, enhances antioxidant capacity and restores mitochondrial function in fulminant hepatitis by promoting MFG-E8 protein translation in an m6A-dependent manner.

Change in volumetric tumor growth rate after cytotoxic therapy is predictive of overall survival in recurrent glioblastoma.

Background: Alterations in tumor growth rate (TGR) in recurrent glioblastoma (rGBM) after treatment may be useful for identifying therapeutic activity. The aim of this study was to assess the impact of volumetric TGR alterations on overall survival (OS) in rGBM treated with chemotherapy with or without radiation therapy (RT). Methods: Sixty-one rGBM patients treated with chemotherapy with or without concomitant radiation therapy (RT) at 1st or 2nd recurrence were retrospectively examined. Pre- and post-treatment contrast enhancing volumes were computed. Patients were considered "responders" if they reached progression-free survival at 6 months (PFS6) and showed a decrease in TGR after treatment and "non-responders" if they didn't reach PFS6 or if TGR increased. Results: Stratification by PFS6 and based on TGR resulted in significant differences in OS both for all patients and for patients without RT (P < 0.05). A decrease of TGR (P = 0.009), smaller baseline tumor volume (P = 0.02), O6-methylguanine-DNA methyltransferase promoter methylation (P = 0.048) and fewer number of recurrences (P = 0.048) were significantly associated with longer OS after controlling for age, sex and concomitant RT. Conclusion: A decrease in TGR in patients with PFS6, along with smaller baseline tumor volume, were associated with a significantly longer OS in rGBM treated with chemotherapy with or without radiation. Importantly, all patients that exhibited PFS6 also showed a measurable decrease in TGR.

Elemental migration and transformation during hydrothermal liquefaction of biomass.

Over the past few decades, energy and environmental crises have worsened due to the excessive consumption of fossil fuels. Hydrothermal liquefaction (HTL) is a promising technology for sustainable biocrude production from biomass. However, elemental migration and transformation during HTL of biomass have only received scant attention to date. Understanding the transformation mechanism is beneficial for downstream biocrude upgrading and by-products utilization for the future industrialization of HTL. In this paper, biomass is grouped into six categories: microalgae, macroalgae, lignocellulose, food waste, manure, and sludge. The biochemical composition and HTL product distribution of six kinds of biomass are compared. The conversion process of the biomacromolecules (including lipids, proteins, cellulose, hemicellulose, and lignin) and the interactions between them are also reported. Furthermore, the distribution of carbon, nitrogen, sulfur, and inorganic elements (Na, K, Ca, Mg, Al, Fe, Zn, Cu, Pb, Cd, etc.) in the HTL products is summarized, and the transformation of the organic and inorganic elements during HTL of biomass is explored. Finally, outlooks for the HTL of biomass are proposed.

Construct a novel anti-bacteria pool from hydrothermal liquefaction aqueous family.

Hydrothermal liquefaction aqueous phase (HTL-AP) is complex and toxic, which severely hinders the scale-up of HTL technology. Distinguished from degrading organics and extracting chemical energy or nutrients from HTL-AP via biological fermentation or algae cultivation, here, we propose an innovative strategy to valorize the HTL-AP as a powerful anti-bacterial pool. Six model ingredients, i.e. lipids, cellulose, xylan, lignin, protein and the mixture were employed, to obtain a thirty-HTL-AP pool for characteristics database construction. We found that the xylan group at 230 °C on Escherichia coli (E. coli) and at 200 °C on Staphylococcus aureus (S. aureus) exhibited the highest anti-bacterial activities via plate experiments, nearly equal to 100 µg/ml streptomycin which far exceeded the working concentration of streptomycin (10-50 µg/ml). The liquid cultivation studies further revealed HTL-APs from the mixture feedstock, protein, real biomass microalgae and cornstalk had more stable anti-bacterial activities as chemically stable substances. Interestingly, the Gram-positive strain S. aureus was more susceptible than the Gram-negative E. coli on the HTL-APs, probably owing to the outer selectively permeable membrane difference and the strong reducibility and acidity of HTL-APs. This study provides a new vision to seek the anti-bacterial potential of HTL aqueous, supporting further investigations on its molecular mechanism and new bactericide development.

Understand the antibacterial behavior and mechanism of hydrothermal wastewater.

Unlocking the antibacterial potential is an emerging strategy to valorizing the toxic wastewater from hydrothermal liquefaction (HTL). Here, we investigated the response and biological mechanism of antibacterial properties of HTL wastewater. Four different biowastes i.e. microalgae, cornstalk, cow manure and swine manure were used as the feedstock of HTL to create wastewater with diverse molecule spectrum, whereas ten strains i.e. five gram-positive strains and five gram-negative strains were employed to represent typical pathogenic microorganism. HTL wastewater exhibited antibacterial potential and obvious reduction on cell viability at high inclusion ratio, although the minimum inhibitory concentration (MIC) and cell response intensity varied depending on different HTL feedstocks and strain species. The decreased ATP generation and increased H2O2 accumulation in treated cells further confirmed the inhibition of HTL wastewater on the cell metabolism. The antibacterial mechanism of HTL wastewater was confirmed, including damage to biomolecules or membranes, depletion of crucial components, disruption of metabolic circuits and imbalance of creation of redox cofactor. The complex compounds in HTL wastewater were probably attributed to the multiple inhibition pathways and the relationship among those multiple pathways was speculated. The present study contributes to the mechanism analysis of complex compound mixture and bactericide characteristics of HTL wastewater.

Glucose metabolism controls human γδ T-cell-mediated tumor immunosurveillance in diabetes.

Patients with type 2 diabetes mellitus (T2DM) have an increased risk of cancer. The effect of glucose metabolism on γδ T cells and their impact on tumor surveillance remain unknown. Here, we showed that high glucose induced Warburg effect type of bioenergetic profile in Vγ9Vδ2 T cells, leading to excessive lactate accumulation, which further inhibited lytic granule secretion by impairing the trafficking of cytolytic machinery to the Vγ9Vδ2 T-cell-tumor synapse by suppressing AMPK activation and resulted in the loss of antitumor activity in vitro, in vivo and in patients. Strikingly, activating the AMPK pathway through glucose control or metformin treatment reversed the metabolic abnormalities and restored the antitumor activity of Vγ9Vδ2 T cells. These results suggest that the impaired antitumor activity of Vγ9Vδ2 T cells induced by dysregulated glucose metabolism may contribute to the increased cancer risk in T2DM patients and that metabolic reprogramming by targeting the AMPK pathway with metformin may improve tumor immunosurveillance.

The hydrochar activation and biocrude upgrading from hydrothermal treatment of lignocellulosic biomass.

The production of hydrochar and biocrude from hydrothermal treatment of lignocellulosic biomass is getting increasing attention, but the quality of hydrochar and biocrude need further improvement before utilization. Many attempts have been carried out on the hydrochar activation and biocrude upgrading. However, different methods play different roles on the property of hydrochar and biocrude, this topic received scant attention in recent review papers. Therefore, the influence of different activation methods on hydrochar property, and the potential application of hydrochar were summarized in this study. Meanwhile, the research progress on biocrude upgrading is reported. Besides, the techno-economic analysis of hydrochar and biocrude from hydrothermal treatment of lignocellulosic biomass are also discussed. Finally, the research needs and future directions on hydrochar activation and biocrude upgrading were proposed. This paper could provide insights for further studies on the utilization of hydrochar and biocrude.

Reduced SULT2B1b expression alleviates ox-LDL-induced inflammation by upregulating miR-148-3P via inhibiting the IKKß/NF-κB pathway in macrophages.

Atherosclerosis is a lipid-driven chronic inflammatory disease in which lipid-laden macrophage foam cells lead to inflamed lesions in arteries. Previous studies have proven that sulfotransferase 2B1b (SULT2B1b) has several roles in the regulation of lipid metabolism and the inflammatory response. However, little is known about the functions of SULT2B1b in ox-LDL-induced inflammation in macrophages. In this study, after treatment with either ox-LDL alone or combined with transfection of siRNAs targeting SULT2B1b, IL-6, TNF-α, NF-κB, IKKß and IκB mRNA and protein expression were determined in Raw264.7 cells by real-time PCR and Western blot, respectively. The proliferative capacity was determined by EdU staining and Cell Counting Kit-8. Our data demonstrated that SULT2B1b knockdown could reduce phosphorylated NF-κB levels and downregulate IKKß protein levels. Additionally, IκB levels were increased and the proliferation of ox-LDL stimulated cells was inhibited after SULT2B1b silencing. Downregulation of SULT2B1b expression was found to upregulate miR-148a-3p expression by microarray assay, while IKKß was a miR-148a-3p target gene. Our study suggests that SULT2B1b knockdown could promote miR148a-3p expression and inhibit activation of the IKKß/NF-κB signalling pathway, which suppressed the inflammatory response in macrophages. Therefore, targeting the SULT2B1b gene might be potentially beneficial for atherosclerosis prevention by decreasing the inflammatory response.

GIPC-Regulated IGFBP-3 Promotes HSC Migration In Vitro and Portal Hypertension In Vivo Through a ß1-Integrin Pathway.

BACKGROUND & AIMS: Transforming growth factor (TGF-ß)-induced activation of quiescent hepatic stellate cells (HSCs) and their transformation to myofibroblasts is a key event in liver fibrosis and portal hypertension. GIPC (also referred to as synectin) is a downstream signal activation molecule of TGF-ß and other receptors. In this study, we sought to identify novel genes targeted by TGF-ß and GIPC and elucidate if and how they may contribute to liver fibrosis. METHODS: We performed sequential messenger RNA sequencing analysis on TGF-ß-stimulated HSCs and then on TGF-ß-stimulated HSCs in the presence and absence of GIPC also referred to as synectin (GIPC) knockdown. Insulin-like growth factor binding protein-3 (IGFBP-3) transport protein emerged as a top activation target of both TGF-ß and GIPC. Quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, targeted chromatin immunoprecipitation, and Western blot analysis were done for further confirmation. RESULTS: IGFBP-3, an insulin growth factor transport protein, emerged as a top activation target of both TGF-ß and GIPC, which was confirmed by quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blot analysis. Targeted chromatin immunoprecipitation showed that GIPC increases the histone 3 lysine 27 (H3K27) acetylation activating mark and concurrently decreases the H3K27 inhibitory trimethylation (H3K27m3) mark, providing an epigenetic correlate to the gene regulation changes. In vivo, global knockout of IGFBP-3 mice resulted in attenuation of HSC activation markers and attenuation of portal pressure in response to chronic liver injury models. Analysis of serum levels from cirrhotic patients also showed an IGFBP-3 increase of more than 2-fold compared with healthy controls. Finally, in vitro mechanism studies showed that IGFBP-3 promotes HSC migration through integrin-dependent phosphorylation of protein kinase B. CONCLUSIONS: TGF-ß up-regulates IGFBP-3 through GIPC, leading to increased HSC migration in vitro and promotes portal hypertension in vivo. These studies support the role of IGFBP-3 as a potential pathophysiologic target or biomarker in chronic liver disease.