CENPB promotes the proliferation of hepatocellular carcinoma and is directly regulated by miR-29a.

Hepatocellular carcinoma (HCC) is a significant global health concern as it ranks as the sixth most common malignant tumor and the third leading cause of cancer-related deaths. In this study, we analyzed the expression of centromere protein B (CENPB) mRNA in HCC using TCGA and GEO datasets. Immunohistochemistry (IHC) was performed to determine CENPB protein levels in 490 HCC patients. Our findings revealed higher expression of CENPB mRNA in HCC tissues across the three datasets. Additionally, as the pathological stage and histological grade advanced, CENPB expression increased. Patients with elevated levels of CENPB mRNA and protein demonstrated shorter overall survival (OS) and recurrence-free survival (OS). Notably, CENPB protein showed prognostic value in patients with stage I/II, AFP levels below 400 ng/ml, and tumor size less than 5 cm. Using multivariate regression analysis in 490 HCC patients, we developed nomograms to predict 1-year, 3-year, and 5-year OS and RFS. Knockdown of CENPB in Hep3B and MHCC97 cell lines resulted in significant inhibition of cell proliferation and invasion. Furthermore, bioinformatics analysis identified miR-29a as a potential negative regulator of CENPB expression, which was validated through a dual-luciferase reporter assay. In conclusion, our findings suggest that CENPB may serve as an oncogenic factor in HCC and is directly regulated by miR-29a, highlighting its potential as a promising therapeutic target.

Ultrabroad-spectrum, multidrug resistant bacteria-killing, and biocompatible quaternized chitin derivative for infected wound healing.

Wound infections have consistently been recognized as serious threats to human. The design of antimicrobial and biocompatible wound dressings for infected wounds is an area of constant research. Herein, we homogeneously synthesized an ultrabroad-spectrum antimicrobial and biocompatible quaternized chitin derivative (QC-4) in a high-efficiency and sustainable route using aqueous KOH/urea solution. Particularly, QC-4 displayed powerful multidrug resistant bacteria-killing activities even at a very low antimicrobial concentration range from 500 ng/mL to 5 µg/mL, including clinically prevalent multidrug-resistant Escherichia coli (MDR-E. coli), methicillin resistant Staphylococcus aureus (MRSA), multidrug-resistant Pseudomonas aeruginosa (MRPA), and multidrug-resistant Acinetobacter baumannii (MDR-A. baumannii). With the aim to facilitate clinical translation, we validated the biocompatibility and safety of QC-4 both in vitro and in vivo, and further assessed the effects of QC-4 on infected wound healing in a porcine infectious full-thickness skin wound model. QC-4 demonstrated significant reduction of microbial aggregates and enhanced wound-healing effects by promoted re-epithelialization and collagen deposition, which were quite comparable to that of commercial Alginate-Ag dressing and absolutely superior to commercial Chitoclot Bandage dressing. Additionally, we provided clear evidences that QC-4 had a unique mechanism of action by attracting electrostatically to the negatively charged microbial surface, thus damaging the microbial cell wall and membrane. Findings of this work provided robust preclinical rationale for the future translational applications of QC-4 as a novel ultrabroad-spectrum and multidrug resistant bacteria-killing antimicrobial wound dressing for clinical wound management.

Hypothermic oxygenated machine perfusion alleviates liver injury in donation after circulatory death through activating autophagy in mice.

Hypothermic oxygenated machine perfusion (HOPE) is a safe and reliable method that could alleviate liver injury in donation after circulatory death (DCD). This study focuses on the role of autophagy in HOPE's protective effect on DCD liver injury. A 30-minute warm ischemic liver model was established in mice. After 4 hours of cold storage (CS), 1 hour of hypothermic machine perfusion (HMP) with 100% O2 or 100% N2 was employed. During 2 hours of reperfusion, liver tissue and perfusate were collected to evaluate liver function, oxidative stress level, apoptosis, and necrosis. Western blotting was used to explore the level of autophagy. When the liver experienced warm ischemic injury, LC3B-II expression was significantly enhanced. Compared with the CS, HOPE induced lower release of AST and ALT, as well as lower oxidative stress levels, apoptosis, and necrosis cell numbers, and led to higher tissue ATP content. Meanwhile, expression of autophagy-related proteins, such as ULK1, Atg5, and LC3B-II, increased. When oxygen was completely replaced by nitrogen, the washout effect of HMP did not activate autophagy and did not relieve DCD liver injury. When the autophagy inhibitor 3-methyladenine was used in HOPE, the protective effect of HOPE was attenuated. In conclusion, DCD liver injury activated autophagy compared with healthy liver, while HOPE alleviated DCD liver injury by increasing autophagy levels further in this mouse model. However, HMP with 100% of N2 had no beneficial effect on DCD liver injury or on autophagy levels compared with CS. The research on autophagy may provide a new strategy for alleviating DCD liver injury in clinical practice.

Hypothermic Oxygenated Machine Perfusion Alleviates Donation After Circulatory Death Liver Injury Through Regulating P-selectin-dependent and -independent Pathways in Mice.

BACKGROUND: Hypothermic oxygenated machine perfusion (HOPE) has been shown to improve the quality of liver donation after circulatory death (DCD) compared to cold storage (CS). However, the mechanism by which HOPE works is unclear. In this study, a mouse liver HOPE system was developed to characterize the role of P-selectin in the protective effect of HOPE on DCD livers. METHODS: A warm ischemia model of the liver and an isolated perfused liver system were established to determine a suitable flow rate for HOPE. Perfusate and tissue samples from wild-type and P-selectin knockout (KO) mice were used to determine liver function, apoptosis and necrosis rates, deoxyribonucleic acid injury and oxidative stress levels, leukocyte and endothelial cell activation, and inflammatory reactions. RESULTS: A mouse liver HOPE system was successfully established. HOPE at flow rates between 0.1 and 0.5 mL/min · g were shown to have a protective effect on the DCD liver. P-selectin KO improved the quality of the DCD liver in the CS group, and reduction of P-selectin expression in the wild-type HOPE group had similar protective effects. Moreover, there was a reduction in the degree of oxidative stress and deoxyribonucleic acid injury in the P-selectin KO HOPE group compared with the P-selectin KO CS group. CONCLUSIONS: We established a mouse HOPE system and determined its suitable flow. We also proved that P-selectin deficiency alleviated DCD liver injury. HOPE protected the DCD liver through regulating P-selectin-dependent and -independent pathways.

HSP27 promotes epithelial-mesenchymal transition through activation of the ß-catenin/MMP3 pathway in pancreatic ductal adenocarcinoma cells.

BACKGROUND: The precise role of heat shock protein 27 (HSP27), as a type of small molecular protein in HSPs, in pancreatic ductal adenocarcinoma (PDAC) remains to be elucidated. The aim of the present study was to investigate the expression and function of HSP27 in PDAC cells. METHODS: We first detected the expression of HSP27 in PDAC tissues. Combining with the clinical pathology characteristics of PDAC patients, the relationship between them was analyzed. Then, we knocked down HSP27 using short interfering RNA (siRNA) and observed its biological functions using scratch assay and matrigel invasion and migration assays in ASPC-1 and PANC-1 cells. In mechanism, we verified the ß-catenin/MMP-3 pathway associated proteins in ASPC-1 and PANC-1 cells. RESULTS: We found that HSP27 was highly expression in PDAC tissues, and was positively correlated with tumor differentiation, TNM staging and poor prognosis of PDAC patients. In vitro, we down-regulated the expression of HSP27 in ASPC-1 and PANC-1 cells and found that the invasion and migration ability of PDAC cells were significantly depressed, meanwhile, the activation of the ß-catenin/MMP-3 pathway was inhibited. CONCLUSIONS: HSP27 may be used as a tumor biomarker for diagnosis of PDAC, and HSP27 can promote the invasion and migration of PDAC by activating the ß-catenin/MMP3 Pathway. Therefore, inhibition of HSP27 has therapeutic potential for the treatment of PDAC.

Green Fabrication of Amphiphilic Quaternized ß-Chitin Derivatives with Excellent Biocompatibility and Antibacterial Activities for Wound Healing.

Bacterial infection has always been a great threat to public health, and new antimicrobials to combat it are urgently needed. Here, a series of quaternized ß-chitin derivatives is prepared simply and homogeneously in an aqueous KOH/urea solution, which is a high-efficiency, energy-saving, and "green" route for the modification of chitin. The mild reaction conditions keep the acetamido groups of ß-chitin intact and introduce quaternary ammonium groups on the primary hydroxyl at the C-6 position of the chitin backbone, allowing the quaternized ß-chitin derivatives (QCs) to easily form micelles. These QCs are found to exhibit excellent antimicrobial activities against Escherichia coli, Staphylococcus aureus, Candida albicans, and Rhizopus oryzae with minimum inhibitory concentrations (MICs) of 8, 12, 60, and 40 µg mL-1 , respectively. As a specific highlight, their inherent outstanding biocompatibility and significant accelerating effects on the healing of uninfected, E. coli-infected, and S. aureus-infected wounds imply that these novel polysaccharide-based materials can be used as dressings for clinical skin regeneration, particularly for infected wounds.

Homogeneous synthesis of quaternized chitin in NaOH/urea aqueous solution as a potential gene vector.

Water-soluble quaternized chitins (QCs) were homogeneously synthesized by reacting chitin with (3-chloro-2-hydroxypropyl) trimethylammonium chloride (CHPTAC) in 8wt% NaOH/4wt% urea aqueous solutions. The chemical structure and solution properties of the quaternized chitins were characterized by (1)H NMR, FT-IR, elemental analysis, dynamic light scattering (DLS) and zeta potential measurements. The results demonstrated that the water-soluble QCs, with a degree of substitution (DS) values of 0.27-0.54, could be obtained by varying the concentration of chitin, the molar ratio of CHPTAC to chitin unit, and the reaction time at room temperature (25°C). Two QCs (DS=0.36 and 0.54) were selected and studied as gene carriers. Agarose gel retardation assay revealed that both QCs could condense DNA efficiently when N/P ratio>3. The results of particle size and zeta potential indicated that both QCs had a good ability of condensing plasmid DNA into compact nanoparticles with the size of 100-200nm and zeta potential of +18 to +36mV. Compared to polyethylenimine (PEI, 25kDa), the QCs exhibited outstanding low cytotoxicity. Transfection efficiencies of the QCs/DNA complexes were measured using pGL-3 encoding luciferase as the foreign DNA, and the QCs/DNA complexes showed effective transfection efficiencies in 293T cells. These results revealed that the QCs prepared in NaOH/urea aqueous solutions could be used as promising non-viral gene carriers owing to their excellent characteristics.