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Background Nonalcoholic fatty liver disease (NAFLD) is a common metabolic disease that affects 20–30% of individuals worldwide. Liver puncture remains the gold standard for the diagnosis of liver diseases despite limitations regarding invasive nature and sample variability. It is of great clinical significance to find noninvasive biomarkers to detect and predict NAFLD. Objective The aims of this study were to identify potential serum markers in individuals with early-stage NAFLD and to advance the mechanistic understanding of this disease using a high-throughput mass spectrometry-based untargeted metabolomics approach. Methods One hundred and twelve patients with early-stage NAFLD aged 18–55 were recruited according to the guidelines. The control group included 112 healthy participants. The demographic, anthropometric, clinical and laboratory data of all participants were systematically collected. Serum samples were obtained after an overnight fast. The comprehensive serum metabolomic analysis was performed by ultra-performance liquid chromatography-Orbitrap mass spectrometry. The resultant data was processed by Compound Discover and SIMCA-P software to validate the potential biomarkers. Significantly altered metabolites were evaluated by variable importance in projection value (VIP > 1) and ANOVA (p < 0.01). Pathway analysis was performed using MetaboAnalyst 4.0. Results The liver function test of early NAFLD patients showed no statistical differences to control group (p > 0.05). However, obvious differences in blood lipids were observed between subjects with NAFLD and controls (p < 0.001). In total, 55 metabolites showed significant changes in experimental group were identified. The area under curve (AUC) values deduced by receiver operating curve (ROC) analysis indicated that these newly identified biomarkers have high predictability and reliability. Of these, 15 metabolites with AUC greater than 0.9 were of great diagnostic value in early NAFLD patients. Conclusion In this study, a total of 15 serum metabolites were found to strongly associate with early NAFLD. These biomarkers may have great clinical significance in the early diagnosis of NAFLD, as well as to follow response to therapeutic interventions.

The aim of this study was to prepare levofloxacin-loaded chitosan microspheres and to evaluate their in vitro and in vivo characteristics. Glutaraldehyde-crosslinked microspheres were prepared using a spray-drying method, and characterized in terms of the morphological examination, particle size distribution, entrapment efficiency, drug loading and in vitro release. Pharmacokinetics and colon biodistribution studies were used to evaluate that microspheres have more advantage than the conventional formulations. The surface morphology of the freeze-dried microspheres were smooth, discrete with a regular spherical to near-spherical shape. Size of the microspheres after freeze-drying was 4.96 ± 0.76 mm and well-distributed. The zeta potential of microspheres was À29.3 ± 2.1 mV. An average drug loading of 9.3 ± 0.4% and encapsulation efficiency of 81.1 ± 4.7% of levofloxacin microspheres were obtained with the optimized preparation parameters. The cumulative release rate of levofloxacin microspheres was followed by a sustained release and fitted for classic Higuchi kinetic model. In vivo studies showed that chitosan microspheres are thought to have the potential to maintain levofloxacin concentration within target ranges for a long time, decreasing side effects caused by concentration fluctuation, ensuring the efficiency of treatment and improving patient compliance by reducing dosing frequency. It also does not cause any harmful or toxic effect in colon and rectum as evaluated by histopathologic studies.

BackgroundAs DNA demethylation protein, Ten-eleven translocation 1 (TET1) has been widely reported that is related to tumorigenesis and tumor metastasis. This study is to investigate the role and regulation mechanism of TET1 in colon cancer.Methods The TET1 and Catenin beta-1 (CTNNB1) expression level in colon cancer samples and cancer cell lines HCT116/SW480 were observed to discover the relationship between these two genes. Knockdown and overexpression of TET1 through shRNA and CRISPR technology were used to elucidate the effect of TET1 on WNT/β-catenin pathway. The 5-hmC/5-mC level were explored by bisulfate sequencing (BSP) and Chromatin immunoprecipitation (ChIP) to further explain the regulation mechanism. Combined with the reverse assay and transwell invasion assay, the cell migration and invasion ability were tested. Finally, the role of TET1 on DOX resistance was analyzed.Results TET1 downregulated in colon cancer and showed an opposite expression trend with WNT pathway associated gene CTNNB1. TET1 bound to CTNNB1 promotor and catalyzed demethylation to activate transcription of CTNNB1, inhibiting WNT/β‐catenin signaling pathways. Colon cancer cells proliferation was promoted by TET1 downregulation, which was further verified as shTET1 could upregulate the tumor invasion. The DOX addition could rescue the cell migration, compared with normal expression of TET1. Meanwhile, TET1 down-regulation was related to DOX resistances.Conclusion TET1 played as a DNA hydroxymethylation activates inhibitors of the WNT/β-catenin signaling pathway in colon tumor and TET1 down-regulation contributed to DOX-resistance, which might provide reference to targeting therapy in clinical practice.