Alteration of serum bile acid profiles of HBV-related hepatocellular carcinoma identified by LC-MS/MS.

BACKGROUND: Hepatocellular carcinoma closely related to metabolic disorders is a common and aggressive liver malignancy. The dysregulation of bile acid homeostasis has emerged as a key factor for the development and progression of HCC. We aimed to investigate the relationship between bile acids and HCC diagnosis and progression. METHODS: A total of 744 HBV-related patients (including 396 HCC patients and 348 patients with chronic liver diseases) were enrolled in the current study. The baseline characteristics of patients were collected from electronic medical records, and the levels of bile acid profiles were determined by LC-MS/MS. Propensity score matching analysis was conducted to reduce the effect of selection bias, and receiver operating characteristic analysis was performed to evaluate the clinical application values of bile acid. RESULTS: Significant differences were observed for most characteristics between the HCC group and the CLD group before PSM analysis. Patients with HCC were older and fatter (p < 0.05). After adjusting with a 1:1 ratio for age, gender and BMI, 42 HCC patients and 42 non-HCC patients were matched in 2 groups, respectively. The total bile acid level in HCC patients was lower than that in patients with chronic liver diseases before and after PSM analysis (p < 0.05). However, patients with HCC had significantly higher levels of DCA, LCA, and GLCA and lower levels of TCDCA, GUDCA, and TUDCA (p < 0.05, respectively). Besides, the TCDCA, TUDCA, GLCA, and GUDCA were significantly correlated with tumor procession. Moreover, the BAs profiles had a superior predictive ability for predicting the development of HCC even in patients with low serum AFP levels. CONCLUSION: Patients with HCC had significantly lower levels of total bile acid, but higher levels of secondary bile acids (DCA, LCA, and GLCA). The levels of primary bile acid (TCDCA) were closely related to tumor size and stage, which indicated that the bile acids were involved in the HCC procession and had important clinical application values.

Runx1 Deficiency Promotes M2 Macrophage Polarization Through Enhancing STAT6 Phosphorylation.

Our previous study had demonstrated that Runx1 promoted LPS-induced macrophage inflammatory response, however, the role of Runx1 in M2 macrophage polarization still remains largely unknown. This study was conducted to investigate the role of Runx1 in IL-4/IL-13-induced M2 macrophage polarization and its potential regulatory mechanism. We found that exposure of macrophages to IL-4/IL-13 induced a remarkable increasement in Runx1 expression level. Specifically, we established genetically modified mice lacking Runx1 in myeloid cells, including macrophages. RNA-Seq was performed to identify differentially expressed genes (DEGs) between Runx1 knockout and WT control bone marrow-derived macrophages (BMDMs). We identified 686 DEGs, including many genes which were highly expressed in M2 macrophage. In addition, bioinformatics analysis indicated that these DEGs were significantly enriched in extracellular matrix-related processes. Moreover, RT-qPCR analysis showed that there was an obvious upregulation in the relative expression levels of M2 marker genes, including Arg1, Ym1, Fizz1, CD71, Mmp9, and Tgm2, in Runx1 knockout macrophages, as compared to WT controls. Consistently, similar results were obtained in the protein and enzymatic activity levels of Arg1. Finally, we found that the STAT6 phosphorylation level was significantly enhanced in Runx1 knockout macrophages, and the STAT6 inhibitor AS1517499 partly reduced the upregulated effect of Runx1 deficiency on the M2 macrophage polarization. Taken together, Runx1 deficiency facilitates IL-4/IL-13-induced M2 macrophage polarization through enhancing STAT6 phosphorylation.

PDGF regulates chondrocyte proliferation through activation of the GIT1- and PLCγ1-mediated ERK1/2 signaling pathway.

Studies investigating the effects of cytokines on chondrocytes have significant application potential, since the culture of cartilage cells in vitro is a vital step for cartilage tissue engineering. Platelet-derived growth factor (PDGF), one of the growth factors occurring at the early stage of the healing process of damaged tissue, is critical in bone healing. The present study investigated the effects of the activation of PDGF on cell proliferation, apoptosis and the underlying mechanisms of chondrocytes in vitro. The results indicated that the stimulation of PDGF led to overexpression of the G-protein-coupled receptor kinase interacting protein-1 (GIT1) and promotion of the phosphorylation of phospholipase Cγ1 (PLCγ1). Furthermore, PDGF induced chondrocyte proliferation and inhibited apoptosis via activation of the extracellular signal-regulated kinase (ERK) 1/2 pathway. Following knocking down GIT1 expression by small interfering RNA, phosphorylation of PLCγ1 and activation of the ERK1/2 pathway was no longer promoted by PDGF. In addition, the effects of PDGF on proliferation and apoptosis were suppressed. The expression levels of GIT1 were not affected; however, the phosphorylation of ERK1/2 was suppressed through inhibition of the phosphorylation of PLCγ1 by U73122. The results demonstrated that GIT1 is upstream of PLCγ1. Although the ability of PDGF to induce cell proliferation was inhibited by the inhibition of the ERK1/2 pathway by PD98059, apoptosis was not suppressed. In conclusion, the present study demonstrated that PDGF was able to activate the GIT1­PLCγ1­mediated ERK1/2 pathway to control chondrocyte proliferation.

Regulation of chondrocyte proliferation through GIT1-Rac1-mediated ERK1/2 pathway by PDGF.

There are many growth factors contributing to fracture healing after bone fractures. Platelet-derived growth factor (PDGF) released from platelets is a factor promoting cell division and proliferation, and first appears around the sites of fractures. Culture of chondrocytes in vitro are stimulated by PDGF to proliferation, its presence being upregulated in the extracellular matrix of cartilage; the main components include aggrecan and type II collagen. PDGF induces the expression of G the protein-coupled receptor kinase interacting protein 1 (GIT1), promoting Rac1 and ERK1/2 phosphorylation. Both knocking down GIT1 expression by siRNA and blocking phosphorylation of Rac1 inhibit this induced proliferation of chondrocyte. GIT1 and Rac1 control each other, having a synergistic effect on activation of the ERK1/2 pathway. The results suggest that PDGF regulates chondrocyte proliferation through activation of ERK1/2 pathway by upregulation of GIT1 expression and Rac1 phosphorylation.

[Determination of silver content in silver-loaded coral hydroxyapatite in relation to the biocompatibility].

OBJECTIVE: To determine the amount of silver in silver-loaded coral hydroxyapatite (Ag(+)-CHA) bone substitute and its impact on the biocompatibility of this material with mouse embryonic osteoblast cells. METHODS: Ag(+)-CHA was prepared by immersing coral hydroxyapatite in a serial concentration of silver nitrate solutions. The amount of silver in the prepared Ag(+)-CHA was measured by inductively coupled plasma atomic emission spectrometry (ICP-AES). The viability of MC3T3-E1 cells incubated with Ag(+)-CHA was measured by MTT colorimetric assay, and the cell growth and morphological changes were observed by inverted phase-contrast microscope and confocal laser scanning microscope. RESULTS: The amount of silver loading in the bone substitutes prepared by immersion in 1×10(-2), 1×10(-3), 5×10(-4), 10(-4), 8×10(-5), and 5×10(-5) mol/L silver nitrate solutions were 4127.67∓47.35, 167.90∓11.00, 83.42∓4.51, 30.20∓2.32, 22.39∓4.09, and 15.11∓0.55 µg/g, respectively. A low silver content in the material (prepared with silver nitrate solution of less than 8×10(-5) mol/L) showed no significant inhibitory effect on the growth of MC3T3-E1 cells or produced noticeable cytotoxic effect. On the materials prepared with 8×10(-5) and 10(-5) mol/L silver nitrate solution, the osteoblasts displayed active proliferation similar to those incubated on materials without silver loading. The confluent cells showed a normal fusiform morphology with tight arrangement. CONCLUSION: Ag(+)-CHA with low silver content has a good biocompability and can promote the proliferation and growth of MC3T3-E1 cells in vitro, suggesting the clinical potential of this material as a anti-infection bone substitute.

[Diagnosis and treatment for complicated atlantoaxial dislocation].

OBJECTIVE: To explore the clinical characteristics and treatment methods for complicated atlantoaxial dislocation. METHODS: A retrospective evaluation was done to summarize and analyze the clinical characteristics and complicated factors of 54 patients with complicated atlantoaxial dislocation who could not to be treated effectively by using conventional therapy in our hospital from February 2005 to October 2008. According to different complicated factors, different treatment methods mainly including transoral atlantoaxial reduction plate-III (TARP-III) operation, decompression procedure with deep grinding guided by computer aided design-rapid prototyping (CAD-RP), screw placement technique with CAD-RP guide plate and extensile approach surgery were performed. RESULTS: The average follow-up period was 24 months. Among 54 cases, 48 cases achieved immediate anatomic reduction completely and 6 cases almost achieved anatomical reduction. All the compressed spinal cords were decompressed sufficiently. The decompression rate was 86.0% and the improvement rate of nerve function was 77.8%. Two cases suffered postoperative intracranial infection. CONCLUSION: Some cases of complicated atlantoaxial dislocation can be effectively treated by using TARP-III operation, decompression procedure with deep grinding guided by CAD-RP, individualized screw placement technique with CAD-RP guide plate and extensile approach surgery.

Determination of antibacterial properties and cytocompatibility of silver-loaded coral hydroxyapatite.

In this study, silver-loaded coral hydroxyapatites (SLCHAs) were used as scaffolds for bone tissue engineering. The SLCHAs were prepared by surface adsorption process and ion-exchange reaction between Ca(2+) of coral hydroxyapatite (CHA) and Ag(+) of silver nitrate with different concentrations at room temperature. The properties of the composite SLCHAs were investigated by inductively coupled plasma-atomic emission spectrometry (ICP-AES), scanning electron microscropy (SEM) equipped with backscattered electron detector (BSE), and energy-dispersive X-ray spectrometer (EDS). The SEM images showed that the morphology of the SLCHAs depended on the content of Ag(+), and the silver ions were uniformly distributed on the surface of SLCHAs. The ICP-AES results demonstrated that the silver content of the SLCHAs decreased along with the decrease of the concentration of silver nitrate. The SLCHAs were found effective against Escherichia coli and Staphylococcus aureus by antibacterial test. Mouse embryonic pre-osteoblast cells (MC3T3-E1) were used to test the cytocompatibility of SLCHAs, CHA, and pure coral. Cell morphology and cell proliferation were studied with SEM, laser scanning confocal microscope (LSCM), and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay after 1, 3, and 5 days of culture. The results indicated the cell morphology and proliferation on the scaffolds of Ag(+) (13.6 microg/ml)/CHA and Ag(+) (1.7 microg/ml)/CHA were better than that on Ag(+) (170 microg/ml)/CHA. In addition, adhesion of MC3T3-E1 on the scaffolds showed that the confluent cells showed fusiform shape and arranged tightly on the scaffolds. All the results showed that the antibacterial SLCHAs would have potential clinical application as the scaffolds for bone tissue engineering.