[Pathogenesis of glucocorticoid-induced osteoporosis based on label-free mass proteomics].

OBJECTIVE: To explore pathogenesis of glucocortocoid-induced osteoporosis(GIOP) based on label-free mass proteomics. METHODS: Twevle female Sprague-Dawley(SD) rats were randomly divided into two groups, named as sham group and GIOP group. After one-week adaptive feeding, the rats of GIOP group were administered with dexamethasone via intramuscular injection according to 2.5 mg/kg weighting, while the rats of sham group were administered with the same amount of saline, twice a week. The tibias of each group were collected after 8-week modeling and made pathological sections to confirm the success of modeling. Three samples of each group were picked up to perform label-free mass proteomics. After quality control, differentially expressed proteins were identified according to qualitative and quantitative analyses. Then gene ontology(GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis, cluster analysis as well as protein-protein interaction analysis were performed using bioinformatics analysis. RESULTS: Compared with sham group, the structure of bone trabecular in GIOP group showed abnormal arrangement, uneven distribution and obvious fragmentation, which could demonstrate successful modeling. A total of 47 differentially expressed proteins (DEPs) were identified including 20 up-regulated and 27 down-regulated proteins. The expression of protein nucleophosmin 1(NPM1), adipocyte plasma membrane associated protein (APMAP), cytochromec oxidase subunit 6A1 (COX6A1) and tartrate-resistant acid phosphatase (ACP5) showed a significant difference between two groups. KEGG results showed DEPs were enriched on metabolism-related pathways, immune-related pathways and AMP-activated kinase (AMPK) signaling pathway. CONCLUSION: Protein NPM1, APMAP, COX6A1 and ACP5 showed a close relationship with pathogenesis of GIOP, which could serve as potential biomarkers of GIOP. AMPK signaling pathway played an important role in the occurrence and development of GIOP, which could be regarded as potential signaling pathway to treatment GIOP.

ZeXie decoction alleviates non-alcoholic fatty liver disease in rats: the study of genes, lipids, and gut microbiotas.

Recently, with increasing awareness of health issues, non-alcoholic fatty liver disease (NAFLD) has become an epidemic attracting global attention. As a serious chronic disease, NAFLD is clinically managed with pharmacological interventions that are usually associated with poor long-term efficacy and adverse effects. In this scenario, traditional Chinese medicine (TCM) characterized by "multiple ingredients-multiple targets-multiple pathways" shows promise as a potential option to treat NAFLD. Zexie decoction (ZXD) is a classical TCM formula that possesses favorable lipid-lowering and anti-inflammatory activities. Accumulating evidence indicates that ZXD displays robust efficacy in treating NAFLD. The effectiveness of ZXD against NAFLD has been evaluated in our previous studies. This study further examines its probable mechanism of action in an in-depth manner using multi-omic analysis based on the gut-liver axis and sheds light on the potential relationship among genes, hepatic lipid metabolites, and gut microbiotas. Totally, 71 differentially expressed genes (34 upregulated and 37 downregulated genes), 31 differential lipid molecules (8 upregulated and 23 downregulated), and 56 differential gut microbiotas (37 upregulated and 19 downregulated) were identified in the ZXD-treated group rats compared with the negative control group rats. Of these, owing to their key role in the association analysis, g_Blautia, g_Romboutsia, and g_Lactobacillus were hypothesized to be crucial gut microbiotas in the ZXD-mediated treatment of NAFLD. These microbiotas were found to synergize with key genes, such as AKR1B8, CCN1, and TNKS2, and hepatic lipid metabolites, such as glycerophospholipid and sphingomyelin, which might play a therapeutic role by regulating fatty acid synthesis, correcting lipid metabolism disorder, or reducing the inflammatory response. Overall, the present study provides fresh insights into the ZXD-mediated treatment of NAFLD, which, in turn, is expected to give a push to the modernization of TCM.

Qualitative Analysis of Drug-Containing Plasma and its Application to Quantitative Analysis and Pharmacokinetic Study of Zexie Decoction Using UPLC-MS/MS.

ZeXie Decoction (ZXD) is one of the traditional Chinese medicine formulas (TCMFs) comprising Alisma orientalis (Sam.) Juzep. (ZX) and Atractylodes macrocephala Koidz. (BZ) in 5:2 ratios and is widely employed in clinical applications since ancient times. In this study, UHPLC-QE-Orbitrap-MS was used for qualitative analysis of ZXD in rats' plasma after a single oral dose of 750 mg/kg body weight. Afterward, UHPLC-Q-TRAP-MS/MS was used for simultaneous analysis of three bioactive chemical compounds including alisol A, alisol B, and alisol A 24-acetate in ZXD's ethanol extract. Subsequently, the pharmacokinetic profiles of the three analytes were investigated in rat plasma utilizing UHPLC-Q-TRAP-MS/MS. The multiple reaction monitoring (MRM) mode for the three analytes were at m/z 508.4→383.2 for alisol A, m/z 490.4→365.2 for alisol B, and m/z 550.4→515.5 for alisol A 24-acetate. The analysis method was validated in terms of its accuracy, stability, repeatability, linearity, spiked recovery and matrix effect. As a result, twenty-five chemical constituents of ZXD were putatively identified in plasma, and rapid, sensitive, and accurate methods were established for the quantitative analysis and pharmacokinetic study of ZXD. The findings of this study can provide a scientific base for further study of in vivo pharmacokinetics of TCMFs.

A Strategy for Screening the Lipid-Lowering Components in Alismatis Rhizoma Decoction Based on Spectrum-Effect Analysis.

Alismatis Rhizoma decoction (ARD), comprised of Alisma plantago-aquatica subsp. orientale (Sam.) Sam and Atractylodes macrocephala Koidz. at a ratio of 5 : 2, is a classic traditional Chinese medicine (TCM) formula with successful clinical hypolipidemic effect. This paper aimed to explore the major bioactive compounds and potential mechanism of ARD in the treatment of hyperlipidemia on the basis of spectrum-effect analysis and molecular docking. Nine ARD samples with varying ratios of the constituent herbs were prepared and analyzed by UPLC-Q-TOF/MS to obtain the chemical spectra. Then, the lipid-lowering ability of the nine samples was tested in an oleic acid-induced lipid accumulation model in human hepatoma cells (HepG2). Grey relational analysis and partial least squares regression analysis were then performed to determine the correlation between the chemical spectrums and lipid-lowering efficacies of ARD. The potential mechanisms of the effective compounds were investigated by docking with the farnesoid X receptor (FXR) protein. The results indicated that alisol B 23-acetate, alisol C 23-acetate, and alisol B appeared to be the core effective components on hyperlipidemia in ARD. Molecular docking further demonstrated that all three compounds could bind to FXR and were potential FXR agonists for the treatment of hyperlipidemia. This study elucidated the effective components and potential molecular mechanism of action of ARD for treating hyperlipidemia from a perspective of different compatibility, providing a new and feasible reference for the research of TCM formulas such as ARD.

Integrating Network Pharmacology and RT-qPCR Analysis to Investigate the Mechanisms Underlying ZeXie Decoction-Mediated Treatment of Non-alcoholic Fatty Liver Disease.

ZeXie Decoction (ZXD) is a traditional Chinese medicine composed of Alisma orientalis (Sam.) Juzep. and Atractylodes macrocephala Koidz. ZXD has been widely used to treat non-alcoholic fatty liver disease (NAFLD). The mechanistic basis for the pharmacological activity of ZXD, however, remains poorly understood. In this study, we used a network pharmacology approach and investigated the association between ZXD and NAFLD. We identified the active ingredients of ZXD and screened the potential targets of these ingredients, after which a database of relevant NAFLD-related targets were constructed and several enrichment analyses were performed. Furthermore, the ethanol and aqueous extracts of ZXD were prepared and experimental pharmacology validation was conducted using RT-qPCR of the non-alcoholic fatty liver disease (NAFLD) model in Sprague-Dawley (SD) rats. As a result, a herb-compound-target-pathway network model was developed, and HMGCR, SREBP-2, MAPK1, and NF-κBp65 targets were validated. The gene expression results of these four targets were consistent with those of the network pharmacology prediction. Using an integration strategy, we revealed that ZXD could treat NAFLD by targeting HMGCR, SREBP-2, MAPK1, and NF-κBp65.

Retraction of "Biological Effects of Titanium Surface Charge with a Focus on Protein Adsorption".

[This retracts the article DOI: 10.1021/acsomega.0c02518.].

Biological Effects of Titanium Surface Charge with a Focus on Protein Adsorption.

The effect of changes in surface charge on the biological properties of implants is not clear. The objective of this study was to evaluate the biological properties of the surface of titanium sheets with different charges due to different treatment methods. Titanium sheets were sandblasted with large grit and underwent acid etching before being subsequently divided into the following groups: SLA, no further treatment; SLA-Ca2+, immersed in 1% CaCl2 solution; SLA-NaCl, immersed in saline; and SLA-Ca2+-NaCl, immersed in 1% CaCl2 solution followed by saline. Surface characteristics were evaluated using field-emission scanning electron microscopy with energy-dispersive spectrometry, surface profilometry, and contact angle assays. Additionally, we used a ζ-potential analyzer to directly measure the electrostatic charge on the different group surfaces. The effect of changes in the Ti surface on biological processes after different treatments was determined by analyzing fibronectin adsorption, osteoblast-like MG63 cell adhesion and proliferation, and the expression of osteogenesis-related genes. Compared to the SLA surface, the other three groups contained corresponding trace elements because they were soaked in different liquids; the contact angles of the three groups were not significantly different, but they were significantly smaller than that of the SLA group; and there was no change in the surface topography or roughness. Furthermore, the SLA-Ca2+ group had a significantly reduced negative charge compared to that of the other three groups. There were no differences between the SLA-NaCl and SLA-Ca2+-NaCl groups in terms of negative charge, and the SLA group surface carried the most negative charge. Fibronectin adsorption capacity and cytological performance testing further showed that the SLA-Ca2+ group had the most significant change, followed by the SLA-NaCl and SLA-Ca2+-NaCl groups; the SLA group had significantly lower capacity and performance than the other three groups. These results suggest that the surface charge of the titanium sheet changed when immersed in different liquids and that this treatment enhanced biocompatibility by reducing the electrostatic repulsion between biomaterials and biomolecules.

Evaluation of protein arginine deiminase-4 inhibitor in TNBS- induced colitis in mice.

BACKGROUND AND AIM: Many evidences indicated that neutrophil extracellular traps (NETs) are involved in the pathogenesis of inflammatory bowel disease (IBD). Citrullination of histones by Protein Arginine Deiminase-4 (PAD4) is central for NETs formation. This paper aimed to explore the definite role of NETs in mouse model of Crohn's disease (CD) with 2,4,6-trinitrobenzene sulfonic acid (TNBS). METHODS: The expression of NETs-associated proteins and mRNAs in colon tissue were detected by immunohistochemistry and Real-time Quantitative PCR (QPCR) respectively. Neutrophils were isolated and stimulated in vitro to form NETs. In addition, we also administered Cl-amidine, PAD4 inhibitor, resulting in less NETs formation to investigate protective effect by measuring weight loss, gross bleeding, colon length, myeloperoxidase (MPO) activity, and cytokine expression in mice. RESULTS: The results showed enhanced expression of Ly6G, citrullinated histone H3 (CitH3), and PAD4 in TNBS-induced colitis mice and higher ability of neutrophil to produce NETs in vitro. Blocking NETs formation through Cl-amidine effectively alleviated the clinical colitis index and tissue inflammation in TNBS mice, regulated the expression of pro- or anti-inflammatory cytokines. In addition, Cl-amidine reduced the gene expression of PAD4 and the expression of NETs-associated proteins in the colon of TNBS mice and inhibited the formation of NETs in vitro. CONCLUSIONS: Our data showed that Cl-amidine could alleviate the clinical colitis index in TNBS mice to some extend and suggested blocking NETs formation through inhibition of PAD4 as therapeutic targets for the treatment of CD.

Comparison of the effectiveness and antibiotic cost among ceftriaxone, ertapenem, and levofloxacin in treatment of community-acquired complicated urinary tract infections.

PURPOSE: To study characteristics of patients with community-acquired complicated urinary tract infections (cUTIs) and to compare effectiveness and antibiotic cost of treatment with ceftriaxone (CRO), levofloxacin (LVX), and ertapenem (ETP). METHODS: This retrospective study enrolled patients who had community-acquired cUTIs admitted to Division of Infectious Diseases in a single medical center from January 2011 to March 2013. Effectiveness, antibiotic cost, and clinical characteristics were compared among patients treated with CRO, LVX, and ETP. RESULTS: There were 358 eligible cases, including 139 who received CRO, 128 treated with ETP, and 91 with LVX. The most common pathogen was Escherichia coli. The susceptibilities of these three agents were higher and more superior than first-line antibiotics. Treatment with ETP was associated with a significantly shorter time to defervescence since admission (CRO: 39 hours, ETP: 30 hours, and LVX: 38 h; p = 0.031) and shorter hospitalization stay (CRO: 4 days, ETP: 3 days, and LVX: 4 days; p < 0.001). However, the average antibiotic costs in the CRO group were significantly lower than that in the other two groups [CRO: 62.4 United States dollars (USD), ETP: 185.33 USD, and LVX: 204.85 USD; p < 0.001]. CONCLUSION: The resistance of cUTIs isolates to first-line antibiotic is high. Using ETP, CRO, and LVX in the treatment of cUTIs for good clinical response should be suggested. Among the three agents, ETP had better susceptibility than CRO and LVX, reached defervescence sooner, and was associated with shorter hospital stays. However, using CRO in cUTIs was less expensive than the other two agents.