TRIM21 is critical in regulating hepatocellular carcinoma growth and response to therapy by altering the MST1/YAP pathway.

Liver cancer is the sixth most common cancer and the third leading cause of cancer-related death globally. Despite efforts being made in last two decades in cancer diagnosis and treatment, the 5-year survival rate of liver cancer remains extremely low. TRIM21 participates in cancer metabolism, glycolysis, immunity, chemosensitivity and metastasis by targeting various substrates for ubiquitination. TRIM21 serves as a prognosis marker for human hepatocellular carcinoma (HCC), but the mechanism by which TRIM21 regulates HCC tumorigenesis and progression remains elusive. In this study, we demonstrated that TRIM21 protein levels were elevated in human HCC. Elevated TRIM21 expression was associated with HCC progression and poor survival. Knockdown of TRIM21 in HCC cell lines significantly impaired cell growth and metastasis and enhanced sorafenib-induced toxicity. Mechanistically, we found that knockdown of TRIM21 resulted in cytosolic translocation and inactivation of YAP. At the molecular level, we further identified that TRIM21 interacted and induced ubiquitination of MST1, which resulted in MST1 degradation and YAP activation. Knockdown of MST1 or overexpression of YAP reversed TRIM21 knockdown-induced impairment of HCC growth and chemosensitivity. Taken together, the current study demonstrates a novel mechanism that regulates the Hippo pathway and reveals TRM21 as a critical factor that promotes growth and chemoresistance in human HCC.

Age-specific difference in the temporal relationships between updated cardiovascular health construct and arterial stiffness in middle-aged and older adults.

Previous studies have found that the association between modifiable risk factors and arterial stiffness varied with age. We aimed to explore the age-specific difference in the relationship between new cardiovascular health (CVH) score and arterial stiffness and further detected the age-specific temporal relationships in a prospective cohort study. During a median 4.3 years follow-up, 3757 participants were recruited in this study. A modified AHA "Life's Essential 8" construct (mLE8 with lacking information on diet habits) was used to evaluate CVH. Branchial-ankle pulse wave velocity (baPWV) was measured to assess arterial stiffness. Data were analyzed with logistic regression models, restricted cubic splines (RCS), and cross-lagged path analysis (age < 60 vs. age ≥ 60). In age-stratified analysis, moderate (OR = 2.21, 95% CI 1.11-4.43) and low (OR = 3.37, 95% CI 1.63-7.00) CVH were related with a higher incidence of elevated baPWV compared to high CVH in middle-aged adults, while this association was not detected in older adults. RCS curve showed a steeper linear association between CVH score and elevated baPWV in middle-aged adults than older individuals. In the cross-lagged path analysis, the decline in CVH score preceded the increment in arterial stiffness in middle-aged adults, but they appeared to alter simultaneously in older adults. Our study detected an age-specific difference in the relationship between mLE8 CVH score and elevated baPWV and showed that low CVH preceded alterations of baPWV in middle-aged adults, suggesting the importance of improvement in CVH during the early stage of the lifespan.

Subclinical atherosclerosis associates with diabetic retinopathy incidence: a prospective study.

BACKGROUND: The prospective correlation between subclinical atherosclerosis and diabetic retinopathy (DR) incidence in Chinese patients with type 2 diabetes mellitus (T2DM) remains elusive. METHODS: Prospective data were obtained from 2781 patients with diabetes, among whom 1,964 and 2,180 T2DM patients without any and referable DR at baseline, respectively, were included in the analysis. Multivariate analyses were performed using the Cox proportional hazards model. RESULTS: Over a median follow-up of 22.2 months (interquartile range 12.7-27.7), 282 (14.36%) and 125 (5.73%) patients developed any and referable DR, respectively. After adjustment for confounders, each standard deviation (SD) increase in brachial-ankle pulse wave velocity (ba-PWV) was associated with 31% (95% confidence interval 1.15-1.50) and 38% (1.14-1.66) higher risks of incident any and referable DR, respectively. Compared with the lowest ba-PWV quartile, the highest ba-PWV quartile had 135% (1.48-3.72) and 293% (1.83-8.44) higher risks of developing any and referable DR, respectively. Per SD increase of pulse pressure (PP) was associated with 22% (1.09-1.38) and 22% (1.02-1.46) higher risks of incident any and referable DR, respectively. The restricted cubic spline models further indicated a significant linear association of baseline subclinical atherosclerosis with referable DR, and a nonlinear association with any DR. In addition, adding the ba-PWV to the prognostic model for DR incidence improved the C-statistic value, the integrated discrimination improvement value, and the net reclassification improvement value (all P < 0.05). CONCLUSIONS: Baseline subclinical atherosclerosis was significantly associated with an increased risk of DR incidence, and elevated ba-PWV independently predicted incident DR in T2DM patients.

METTL3-mediated macrophage exosomal NEAT1 contributes to hepatic fibrosis progression through Sp1/TGF-ß1/Smad signaling pathway.

Hepatic fibrosis (HF) is caused by chronic hepatic injury and is characterized by hepatic stellate cells (HSCs) activation. Studies focusing on the function of exosomes derived from macrophages in HF progression are limited. This study aims to identify the roles of exosomal NEAT1 derived from macrophages on HF and the underlying mechanisms. Our studies showed that METTL3 targeted and enhanced NEAT1 expression in macrophages. Exosomal NEAT1 originating from LPS-treated macrophages promoted HSCs proliferation and migration, and induced the expression of fibrotic proteins including collagen I, α-SMA, and fibronectin. Macrophage exosomal NEAT1 contributed to HSCs activation by sponging miR-342. MiR-342 directly targeted Sp1 and suppressed its downstream TGF-ß1/Smad signaling pathway, which eventually led to the inhibition of HSCs activation. Depletion of NEAT1 in the macrophage exosomes inhibited HF progression both in vitro and in vivo. Altogether, our study proved that silence of NEAT1 in the macrophage exosomes exerted protective roles against HF through the miR-342/Sp1/TGF-ß1/Smad signaling pathway, suggesting a potential therapeutic target in HF treatment.

MOF-assisted antifouling material: application in rapid determination of TB gene in whole-serum specimens.

Biofouling is a nuisance in the practical applications of biosensors, which seriously affects the reliability and accuracy of detection. The utilization of antifouling interface materials is a promising option for mitigating biofouling. Only highly accumulated antifouling polymeric surfaces tend to offer "zero" nonspecific protein adsorption. Herein, superior antifouling coatings based on chondroitin sulfate (CS) were prepared by the NH2-MIL-53 (Al) assisted strategy. This is a novel design to improve the antifouling property of material by taking advantage of the high specific surface area of the three-dimensional MOF to increase the accumulation degree of antifouling functional groups per unit area. And the related chemical technology is simple and easy to operate. As expected, this novel CS-loaded MOF demonstrated an excellent antifouling performance in various biological samples, even in 100% goat serum. Only 8.48% changes of differential pulse voltammetry (DPV) were found. Furthermore, this antifouling interface material is successfully applied for the specific detection of the tuberculosis (TB) gene in undiluted biofluids. This developed TB biosensor showed a high analytical performance with a wide linear range (1.00 × 10-16 M to 1.00 × 10-11 M) and a low detection limit, indicating that it may open new avenues for direct biosensing of disease markers for clinical samples.

The METTL3/MALAT1/PTBP1/USP8/TAK1 axis promotes pyroptosis and M1 polarization of macrophages and contributes to liver fibrosis.

Pro-inflammatory M1 macrophages, via activating hepatic stellate cells, contribute to liver fibrosis. In this study, we examined the mechanism and the significance of a signaling axis, METTL3/MALAT1/PTBP1/USP8/TAK1, in regulating pyroptosis and M1 polarization of hepatic macrophages. Liver fibrosis model was established in vivo by CCl4 treatment; M1 polarization was induced in vitro by treating macrophages with lipopolysaccharide or interferon γ. Expressions of METTL3, MALAT1, PTBP1, USP8, and TAK1 were measured by RT-PCR and/or Western blot in Kupffer cells (KCs) isolated from in vivo model or in vitro activated macrophages. Macrophage phenotypes including inflammation (RT-qPCR analysis of a panel of proinflammatory cytokines and ELISA on productions of interleukin (IL)-1ß and IL-18) and pyroptosis (Western blot of NLRP3, Caspase-1, and GSDMD) were investigated. The impact of METTL3 on m6A methylation of MALAT1 was examined by methylated RNA immunoprecipitation (RIP), the interaction between PTBP1 and MALAT1 or USP8 mRNA by combining RNA pull-down, RIP, and RNA stability assays, and the crosstalk between USP8 and TAK1 by co-immunoprecipitation and protein degradation assays. Functional significance of individual component of METTL3/MALAT1/PTBP1/USP8/TAK1 axis was assessed by combining gain-of-function and loss-of-function approaches. In KCs isolated from in vivo liver fibrosis model or in vitro M1-polarized macrophages, METTL3 was up-regulated, and sequentially, it increased MALAT1 level via m6A methylation, which promoted USP8 mRNA degradation through the interaction with PTBP1. Reduced USP8 expression regulated the ubiquitination and protein stability of TAK1, which promoted pyroptosis and inflammation of macrophages. The signaling cascade METTL3/MALAT1/PTBP1/USP8/TAK1, by essentially stimulating pyroptosis and inflammation of macrophages, aggravates liver fibrosis. Therefore, targeting individual components of this axis may benefit the treatment of liver fibrosis.

HSPB8 promoted intrahepatic cholangiocarcinoma progression by enhancing epithelial-mesenchymal transition and autophagy.

PURPOSE: Heat shock protein B8 (HSPB8) has been recently discovered to be participated in the regulation of tumor progression. However, the function of HSPB8 in intrahepatic cholangiocarcinoma (ICC) has not yet been elucidated. This study studied the function of HSPB8 in ICC progression. METHODS: ICC patients (n = 150) were enrolled. The relationship between clinicopathological characteristics and HSPB8 expression was analyzed. RBE cells were transfected and treated by 3-MA. The RBE cells morphology was observed under a transmission electron microscope. Cell counting kit-8 assay, wound healing assay and Transwell experiment was conducted to detect RBE cells proliferation, migration and invasion. Quantitative reverse transcription-polymerase chain reaction, immunohistochemistry, Western blot and immunofluorescence were used for genes detection in clinical tissues and RBE cells. RESULTS: HSPB8 was up-regulated in ICC tissues than that in adjacent normal tissues. High HSPB8 expression in ICC indicated poor prognosis of patients. HSPB8 expression was mainly expressed in cell cytoplasm and aberrantly increased in RBE cells (P < 0.01). HSPB8 up-regulation promoted RBE cells proliferation, migration and invasion (P < 0.05). HSPB8 down-regulation reduced RBE cells proliferation, migration and invasion (P < 0.01). HSPB8 overexpression facilitated Vimentin expression, LC3-II/LC3-I ratio and inhibited E-cadherin, p62 expression in RBE cells (P < 0.05). Treatment of 3-MA partially reversed HSPB8 promotion on RBE cells proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) (P < 0.05 or P < 0.01). CONCLUSION: HSPB8 promoted ICC progression by enhancing EMT and autophagy. HSPB8 might be an effective target for ICC treatment.

Ppic modulates CCl4-induced liver fibrosis and TGF-ß-caused mouse hepatic stellate cell activation and regulated by miR-137-3p.

Hepatic stellate cell activation, characterized by hyperproliferation and increased release of collagens, is a critical event during the initiation and development of hepatic fibrosis. The deregulated genes among different expression profiles based on online datasets were analyzed, attempting to identify novel potential biomarkers and treatment targets for hepatic fibrosis. The abnormal upregulation of mouse peptidylprolyl isomerase C (Ppic) within the CCl4-caused hepatic fibrosis model in mice was identified according to bioinformatics and experimental analyses. The knockdown of Ppic in the CCl4-caused liver fibrosis murine model significantly improved CCl4-caused liver damage, decreased the fibrotic area, reduced ECM deposition, and reduced the hydroxyproline levels. The knockdown of Ppic in TGF-ß-stimulated mouse hepatic stellate cells inhibited cell proliferation and decreased ECM levels. Through direct targeting, miR-137-3p negatively regulated Ppic expression. Contrastingly to Ppic knockdown, miR-137-3p inhibition further promoted cell proliferation and boosted ECM levels; the effects of miR-137-3p inhibition could be partially reversed by Ppic knockdown. Altogether, mmu-miR-137-3p directly targets Ppic and forms a regulatory axis with Ppic, modulating CCl4-caused hepatic fibrosis in mice and TGF-ß-caused mouse hepatic stellate cell activation.

The miR-139-5p/peripheral myelin protein 22 axis modulates TGF-ß-induced hepatic stellate cell activation and CCl4-induced hepatic fibrosis in mice.

Hepatic stellate cells (HSCs) are the major source of extracellular matrix (ECM)-producing myofibroblasts. When activated by multiple injuries, HSCs become proliferative, contractile, inflammatory and chemotactic and are characterized by enhanced ECM production, which plays a central role in hepatic fibrosis initiation and progression. In the present study, through bioinformatics analysis, we identified the abnormal upregulation of Peripheral Myelin Protein 22 (PMP22) in fibrotic murine liver. In CCl4-induced hepatic fibrosis model in mice and TGF-ß-activated hHSCs, PMP22 was observed remarkably upregulated. In TGF-ß-stimulated hHSCs, PMP22 silencing hindered, whereas PMP22 overexpression aggravated TGF-ß-induced hHSC activation. In CCl4-induced hepatic fibrosis model in mice, PMP22 silencing improved CCl4-caused liver damage and fibrotic changes. Through online tools prediction and experimental validation, miR-139-5p was found to bind to the 3'UTR of PMP22 and negatively regulate the expression of PMP22. In contrast to PMP22 silencing, miR-139-5p inhibition enhanced TGF-ß-induced hHSC activation; the effects of miR-139-5p inhibition on TGF-ß-induced hHSC activation were partially reversed by PMP22 silencing. In conclusion, we identify the abnormal upregulation of PMP22 in TGF-ß-activated HSCs and CCl4-induced hepatic fibrosis model in mice, as well as the pro-fibrotic role of PMP22 through aggravating TGF-ß-induced HSCs activation. miR-139-5p targets the 3'UTR of PMP22 and inhibits PMP22 expression; miR-139-5p hinders TGF-ß-induced HSCs activation through targeting PMP22.

Low fouling strategy of electrochemical biosensor based on chondroitin sulfate functionalized gold magnetic particle for voltammetric determination of mycoplasma ovipneumonia in whole serum.

Based on the successful design and preparation of Fe3O4@Au@polyethylene glycol (PEG)@chondroitin sulfate (CS) nanoparticles, a novel electrochemical biosensor was elaborately developed for MO determination by the drop coating method. This biosensor demonstrates ultra-low fouling properties in complex biological systems, especially in whole serum. Only 2.40% of the differential pulse voltammetry (DPV) signal changes are shown in the resistance test of electrode interface to 100% goat serum. The constructed biosensor also presented a good selectivity (DNA samples for M1, M2 and M3give a response around 14.2-21.4%of that obtained by the target analyte), reproducibility (the relative standard deviation (RSD) is only 4.82%) and storage stability (93.2% of the initial response is maintained after 20 day). Moreover, this biosensor exhibits high analytical performance with a wide line arranges (10-17 M âˆ¼ 10-12 M) in buffer, even in 100% goat serum. In both cases, a low limit of detection (LOD) of 3.3 aM(S/N = 3)is showed. To our knowledge, a few examples reported the application of electrochemical biosensors to the direct ultrahigh sensitive determination of MO, especially in whole serum.

The Development of a Standardized Framework for Primary Nurse Specialists in Diabetes Care in China: A Delphi Study.

BACKGROUND: The number of patients with diabetes has been increasing rapidly according to a 2017 report by the International Diabetes Federation. Diabetes has become one of the most challenging public health problems, and there will be an estimated 143 million patients with diabetes in China by 2035. This puts considerable pressure on nurses who specialize in the care of patients with diabetes in China and increases related social and financial burdens. Clinical practice has proven that strengthening the core competencies of nurses and establishing an evaluation system of core competencies improve both healthcare quality and patient quality of life. However, no core-competence system framework currently addresses the unique characteristics of nurses in China. PURPOSES: The purpose of this study was to construct a core-competence system framework for primary nurse specialists in diabetes care. METHODS: A brainstorming approach was conducted that worked to conceptualize the core competencies of nurse specialists in diabetes care in China. Next, a study group organized this information and conducted a seminar; 50 experts and patients with diabetes were invited to develop the first draft of the framework. Afterward, 50 experts were selected to participate in the Delphi survey. Most indicators were retained after a three-round Delphi process, and the superiority chart was used to determine the weights of the six dimensions. RESULTS: Forty-seven experts completed the consultation. The experts' rate of response ranged from 94% to 100%, the authority coefficient was .91, and the Kendall's coefficients of concordance in Grades 1-3 were .793, .418, and .542, respectively. An increasingly detailed, three-grade system framework was developed, including six first-grade indicators (diabetes professional knowledge, diabetes-related knowledge, communication skills and health education ability, specialized skills, clinical judgment, and specialty development capacity), 23 second-grade indicators, and 87 third-grade indicators. The weights of the six first-grade indicators were .221, .149, .192, .209, .160, and .069, respectively. CONCLUSIONS: The core-competence system framework includes six core competencies, which represent the main characteristics of primary nurse specialists in diabetes care who are highly recommended by experts. It is important to keep in mind that this is only a theoretical framework and thus must be further tested in clinical practice settings in China.

Intensive insulin therapy combined with metformin is associated with reduction in both glucose variability and nocturnal hypoglycaemia in patients with type 2 diabetes.

BACKGROUND: The effect on glucose variability in patients with intensive insulin therapy has not been fully understood. This observational study investigated the different glucose variability and hypoglycaemia patterns in type 2 diabetes patients treated with continuous subcutaneous insulin infusion (CSII) or multiple daily injections (MDI) with or without metformin administration. METHODS: During hospitalization, a total of 501 patients with poor glycaemic control and in initial treatment with either CSII alone (n = 187), CSII + Metformin (n = 81), MDI alone (n = 146), or MDI + Metformin (n = 87) were involved in the final analysis. Data obtained from continuous glucose monitoring were used to assess blood glucose fluctuation and nocturnal hypoglycaemia. RESULTS: Among the 4 groups, no difference was found in mean blood glucose levels. Results in parameters reflecting glucose fluctuation: continuous overlapping net glycaemic action in CSII + Metformin and mean amplitude of glycaemic excursions in MDI + Metformin were significantly lower than those in either CSII alone or MDI alone, respectively, even after adjustment (P = .031 and .006). Frequency of nocturnal hypoglycaemia was significantly decreased in CSII + Metformin as compared with CSII alone (0.6% vs 1.8%) and in MDI + Metformin as compared with MDI alone (1.6% vs 2.3%), with the highest frequency observed in MDI alone and the lowest in CSII + Metformin (all between group P < .001). Consistent results were obtained in between-group comparisons for hypoglycaemia duration. Subgroup analysis matched with baseline body mass index, and glycated haemoglobin and fasting blood glucose further confirmed these findings. CONCLUSION: Metformin added to initial CSII or MDI therapy is associated with a reduction in both glucose fluctuation and nocturnal hypoglycaemic risk in patients with type 2 diabetes.

N'-(3-Bromo-4-methoxy-benzyl-idene)nicotinohydrazide monohydrate.

In the title compound, C(14)H(12)BrN(3)O(2)·H(2)O, the benzene ring is oriented at a dihedral angle of 39.66 (11)° with respect to the pyridine ring. The solvent water mol-ecule links with the organic compound via O-H⋯O, O-H⋯N and N-H⋯O hydrogen bonding.

(E)-N'-(1,3-Benzodioxol-5-ylmethyl-ene)nicotinohydrazide monohydrate.

In the title compound, C(14)H(11)N(3)O(3)·H(2)O, the planar [maximum deviation 0.135 (1) Å] 1,3-benzodioxole ring system is oriented at a dihedral angle of 13.93 (7)° with respect to the pyridine ring. Extensive inter-molecular N-H⋯O, O-H⋯O, O-H⋯N and weak C-H⋯O hydrogen bonding is present in the crystal structure.

N'-(Propan-2-yl-idene)nicotinohydrazide.

Crystals of the title compound, C(9)H(11)N(3)O, were obtained from a condensation reaction of nicotinohydrazide and acetone. In the mol-ecular structure, the pyridine ring is oriented at a dihedral angle of 36.28 (10)° with respect to the amide plane. In the crystal structure, mol-ecules are linked via N-H⋯O hydrogen bonds, forming chains.

(E)-N'-(3,4-Dichloro-benzyl-idene)nicotino-hydrazide monohydrate.

In the title compound, C(13)H(9)Cl(2)N(3)O·H(2)O, the 3,4-dichloro-benzene ring is nearly coplanar with the pyridine ring, making a dihedral angle of 4.78 (8)°. Inter-molecular O-H⋯O, O-H⋯N, N-H⋯O and weak C-H⋯O hydrogen bonding is present in the crystal structure.