The Role of miRNA and Long Noncoding RNA in Cholestatic Liver Diseases.

Cholestatic liver diseases encompass a range of organic damages, metabolic disorders, and dysfunctions within the hepatobiliary system, arising from various pathogenic causes. These factors contribute to disruptions in bile production, secretion, and excretion. Cholestatic liver diseases can be classified into intrahepatic and extrahepatic cholestasis, according to the location of occurrence. The etiology of cholestatic liver diseases is complex, and includes drugs, poisons, viruses, parasites, bacteria, autoimmune responses, tumors, and genetic metabolism. The pathogenesis of cholelstatic liver disease is not completely clarified, and effective therapy is lacking. Clarifying its mechanism to find more effective therapeutic targets and drugs is an unmet need. Increasing evidence demonstrates that miRNA and long noncoding RNA are involved in the progression of cholestatic liver diseases. This review provides a comprehensive summary of the research progress on the roles of miRNA and long noncoding RNA in cholestatic liver diseases. The aim of the review is to enhance the understanding of their potential diagnostic, therapeutic, and prognostic value for patients with cholestasis.

Role of p53 methylation in manganese-induced cyclooxygenase-2 expression in BV2 microglial cells.

Manganese (Mn) is an essential cofactor for many enzymes and plays an important role in normal growth and development. However, excess exposure to manganese (Mn) may be an important environmental factor leading to neurodegeneration. The overexpression of microglial cyclooxygenase-2 (COX-2) plays a key role in neuroinflammation in neurodegenerative diseases. The existing data suggest that Mn can induce neuroinflammation by up-regulating COX-2 expression. However, the mechanisms involved in Mn-induced microglial COX-2 up-regulation remain to be determined. The aim of this study was to investigate the role of p53 in Mn-induced COX-2 expression in microglial cells. The results showed that Mn exposure induced the up-regulation of COX-2 and inhibited the expression of p53 in BV2 microglial cells. The addition of p53 activator and the over-expression of p53 blocked the expression of COX-2 and prostaglandin E2 (PGE2), a COX-2 downstream effector, induced by Mn. Further, Mn increased the methylation of p53 DNA in microglia, while the addition of demethylation reagent 5-Aza-dC enhanced the expression of p53 but decreased the expression of COX-2. These results suggested that Mn may inhibit p53 expression through induction of DNA methylation, which can further induce the expression of COX-2 in microglial cells.

Cesarean section in Shanghai: women's or healthcare provider's preferences?

BACKGROUND: Cesarean section (CS) rate has increased rapidly over the past two decades in China mainly driven by non-medical factors. This study was to compare recalled preferences for CS among first-time mothers in early and late pregnancy with actual delivery mode; to explore factors related to CS preference and CS performed without medical indications; and to consider the role of healthcare providers in delivery mode preferences. METHODS: An anonymous questionnaire survey, combined with data on CS indications taken from the patient record, was conducted among 272 first-time mothers having their first postnatal check-up in one university affiliated obstetrics and gynecology hospital in Shanghai, China, between September 2006 and January 2007. Logistic regression was used to study factors related to the recalled preference for CS and CS performed without medical indication, adjusting for maternal age, education and income. RESULTS: The CS rate was 57% (151/263) among all women, 17% with medical indications and 40% without medical indications. For women without medical indications for CS (n = 215), there was no significant difference between women's preference for CS in early (25%) and late pregnancy (28%); 48% of women actually had CS. Women recalled preferring a vaginal delivery but who had CS were more likely to have had a CS suggested by a prenatal care doctor [OR (95% CI): 20 (3.88-107.1)] or by a delivery obstetrician [OR (95% CI): 26 (6.26-105.8)]. Among women recalled preferring and having CS, a suggestion from the prenatal care doctor to have CS was very common. CONCLUSIONS: In the primiparous women without a medical indication for CS, women recall of a provider suggestion for CS was a strong predictor of CS both among women who recalled a preference for CS and among women who recalled a preference for vaginal delivery. Public health education needs strengthening, including discussion of the risks associated with CS and psychological and social support given to women to help them prepare for and cope with childbirth.

Mannan-conjugated adenovirus enhanced gene therapy effects on murine hepatocellular carcinoma cells in vitro and in vivo.

The incidence of advanced hepatocellular carcinoma (HCC) is increasing worldwide, and its prognosis is extremely poor. For some patients for whom surgical treatments are not appropriate, one can only rely on chemotherapy. In the conventional chemotherapy, side effects usually occurred in most cases due to high toxicity levels. Moreover, the development of drug resistance toward chemotherapeutic agents often prevents the successful long-term use of chemotherapy for HCC. Gene therapy represents the exciting biotechnological advance that may revolutionize the conventional fashion of cancer treatment. Overexpression of phosphatase and tensin homologue (PTEN) in cancer cells carrying deletion/mutant type of it can induce the apoptosis of cancer cells and inhibit cell proliferation. In this work, in order to make full use of the high transfectivity of adenovirus, we managed to conjugate the polysaccharide mannan (polymannose) to the surface of the adenovirus chemically under appropriate oxidizing conditions to prepare the mannan-modified adenovirus (Man-Ad5-PTEN). The cytotoxicity and anticancer activity of Man-Ad5-PTEN were assessed in vitro. Reporter gene expression of LacZ transferred by Man-Ad5-LacZ was verified on mannose receptor-deficient NIH/3T3 cells versus mannose receptor-efficient macrophages. Hepatocellular carcinoma cell lines transduced by mannan-modified adenovirus were assayed for cell cycle, apoptosis, invasion, and migration. Further, we detected the antitumor effect on intraperitoneal H22 tumor-bearing mice treated by Man-Ad5-PTEN alone or combined with chemotherapeutic agent of doxorubicin. The results demonstrated that cell growth suppression was not observed in Chang normal hepatocyte cells and the cell killing by Man-Ad5-PTEN is tumor selective. Further, the results showed that the strategy of mannan conjugation could enhance adenovirus-mediated PTEN gene therapy effects on murine hepatocellular carcinoma cells in vitro and in vivo.

Effect of Optical Wedge Rotary on Ablation Efficiency of Femtosecond Laser on Dental Hard Tissue and Restorative Materials.

Objective: Femtosecond laser (fs-laser) is a novel tooth preparation tool but its ablation efficiency is insufficient. The purpose is to establish a new fs-laser tooth ablation method based on a dual-wedges path ablation system, and explore the efficiency of tooth hard tissue and dental restorative materials ablation. Materials and methods: Extracted third molars, pure titanium, cobalt-chromium alloy, gold alloy, and 3Y-zirconia were prepared into samples. These samples were rotary ablated by an fs-laser with dual-wedges. The wavelength was 1030 nm and the pulse duration was 250 fsec. Laser parameters were set as a repetition frequency of 25 kHz, the power percentages as 50% for dental tissues, and 60% for restorative materials. The optical wedge angle was set as 0°, 20°, 40°, 60°, and 80° for restorative materials, 0°, 20°, 30°, 40°, and 60° for enamel, and 0°, 10°, 20°, 30°, and 40°for dentin. Three times of ablation was processed at each parameter to obtain total 90 ablation microcavities of 6 kinds of materials. The diameter, depth, and volume of microcavities were measured by confocal laser microscopy and plotted against optical-wedge-angle in curves of different materials. One-way analysis of variance (ANOVA) was used to test whether the ablation efficiency between different angles was statistically significant. Results: The ablation efficiency of each material at different optical-wedge-angle was statistically significant (p < 0.05) and tends to be correlated. For dental hard tissue, the enamel ablation efficiency was 208.1 times and dentin ablation efficiency were 65.2 times than before when the wedge angle was 40°. For pure titanium, zirconia, cobalt-chromium, and gold alloys, the ablation efficiencies were 3.1, 10.7, 81.5, and 128.8 times than before when the rotation angle was 80°. Conclusions: The ablation efficiency of dental hard tissues and restorative materials was significantly increased with the increase of laser oblique incidence angle. Clinical Trial Registration number: PKUSSIRB-201949124.

Lipopolysaccharide induces inflammatory microglial activation through CD147-mediated matrix metalloproteinase expression.

Microglia-mediated neuroinflammation plays a vital role in the pathophysiological processes of multiple neurodegenerative diseases. Lipopolysaccharide (LPS) is an environmental poison that can induce inflammatory microglial activation. Matrix metalloproteinases (MMPs) are vital factors regulating microglial activation, and CD147 is a key MMP inducer, which can induce inflammation by inducing MMPs. However, whether it is involved in the regulation of microglial activation has not been reported. In this study, the role of CD147 in LPS-induced microglial inflammatory activation was investigated by establishing in vivo and in vitro models. The results suggested that LPS-induced microglial activation was accompanied by the induction of CD147 expression while the inhibition of CD147 expression could inhibit LPS-induced microglial inflammatory activation. In addition, the results also indicated that the role of CD147 in LPS-induced pro-inflammatory activation of microglia was related to its downstream MMP-3, MMP-8, and autophagy. Furthermore, the inhibition of MMP-3, MMP-8, and autophagy attenuated LPS-induced inflammatory activation of microglia. At the same time, there was a certain interaction between MMPs and autophagy, which is shown that inhibiting the expression of MMPs could inhibit autophagy, whereas inhibiting autophagy could inhibit the expression of MMPs. Taken together, we provided the first evidence that CD147/MMPs can be involved in LPS-induced inflammatory activation of microglia through an autophagy-dependent manner.

In vitro cytocompatibility and osteogenic potential of calcium silicate-based dental cements in a root canal-filling model.

OBJECTIVE: To investigate the in vitro cytocompatibility and osteogenic potential of an experimental calcium silicate-based cement and the inflammatory response in human periodontal ligament stem cells (hPDLSCs). METHODS: Cellular responses, osteogenic-related gene expression, and the production of inflammatory cytokines including interleukin (IL)-6 and IL-8 were studied in hPDLSCs exposed to the experimental root canal-filling material C-Root, the commercial tricalcium silicate-based material BioRoot RCS, and the epoxy resin-based material AH Plus. Differences were analyzed using one-way ANOVA with Bonferroni-adjusted pairwise comparison. RESULTS: Exposure to BioRoot and C-Root caused time-dependent increases in cell proliferation. Significantly more mineralized nodules were formed in cells exposed to AH Plus and BioRoot compared with the negative control. Alkaline phosphatase (ALP) activity was significantly lower in AH Plus cells compared with negative control, BioRoot, and C-Root cells. ALP, osteocalcin (OCN), and runt-related transcription factor2 (RUNX2) mRNA expression levels were all significantly higher in C-Root compared with AH Plus cells at day 7. IL-6 and IL-8 levels differed significantly among the experimental groups, with the highest IL-8 levels in BioRoot cells at days 7 and 14. CONCLUSION: The experimental root canal-filling material C-Root has similar in vitro cytocompatibility to BioRoot and better osteogenic potential than AH Plus.

Predicting Presynaptic and Postsynaptic Neurotoxins by Developing Feature Selection Technique.

Presynaptic and postsynaptic neurotoxins are proteins which act at the presynaptic and postsynaptic membrane. Correctly predicting presynaptic and postsynaptic neurotoxins will provide important clues for drug-target discovery and drug design. In this study, we developed a theoretical method to discriminate presynaptic neurotoxins from postsynaptic neurotoxins. A strict and objective benchmark dataset was constructed to train and test our proposed model. The dipeptide composition was used to formulate neurotoxin samples. The analysis of variance (ANOVA) was proposed to find out the optimal feature set which can produce the maximum accuracy. In the jackknife cross-validation test, the overall accuracy of 94.9% was achieved. We believe that the proposed model will provide important information to study neurotoxins.

[A new approach to protein phosphorylation modification analysis for neuron].

OBJECTIVE: To develop a new method for analysis of protein phosphorylation modification in cultured neuron. METHODS: Cultured neurons were pre-incubated in DMEM without sodium phosphate for 15 min to deplete the metabolic pools. Neurons were then labeled with [32P] orthophosphate (2.78 x 10(6) Bq/ml) for 1.5 h and stimulated by either insulin (100 nmol/L), EGF (20 nm/L) or saline for 0, 5, 20, 60, 120 min. Reactions were terminated by freezing neurons in liquid nitrogen prior to the solubilizing of them in a lysis buffer containing 8 mol/L urea, 4% CHAPS, 2% Bio-lyte, pH 3-10, 2 mmol/L TBP. Protein concentrations were determined with Bio-Rad DC Protein Assay kit. The 32P-labled lysates isoelectrically focused on IPG Drystrip pH 3-10 or pH 4-7 Linear gels were subsequently separated in second-dimensional SDS-PAGE. The dried gel was autoradiographed for 5 days at -70 degrees C with an intensifying screen. Alternatively, the separated proteins were visualized by Coomassie Brilliant Blue (CBB) R250 straining. RESULTS: Autoradiography of the 2-DE-separated 32P-laebled neuron lysates revealed around 100 phosphoproteins. This phosphoprotein pattern was stable at 1.5 h after radiolabelling and did not vary significantly for up to 4 h further incubation in the absence of hormone. Most of the major proteins which are phosphorylated in response to insulin or EGF migrated with pH 4.6-6.5 and MW 20000-130000. Insulin and EGF induced similar but not identical patterns of protein phosphoryltion in neurons. Only a few phosphoproteins were abundant enough to be visualized by CBB straining, suggesting that abundance of these phosphoproteins is extremely low. Responses to both isulin and EGF are marked by more increased labeling of the constitutive phosphoproteins, compared with the appearance of new phosphoproteins. CONCLUSION: This approach co-application of 32P-labeled with 2-DE separation and autoradiography has proven to be specific and sensitive in phosphoprotein analysis for neuron. It was valuable in functional proteomic analysis for protein phosphorylation modification during cellular signal transduction.