MicroRNA 98-5p Overexpression Contributes to Delayed Fracture Healing via Targeting BMP-2.

MicroRNAs (miRNAs) are related to the regulation of bone metabolism. Delayed fracture healing (DFH) is a common complication after fracture surgery. The study attempted to examine the role of miR-98-5p and bone morphogenetic protein (BMP)-2 with the onset of DFH. A total of 140 patients with femoral neck fracture were recruited, including 80 cases with normal fracture healing (NFH) and 60 cases with DFH. MC3T3-E1 cells were induced cell differentiation for cell function experiments. Real-time quantitative polymerase chain reaction (RT-qPCR) was carried out to test mRNA levels. Cell proliferation and apoptosis were determined via CCK-8 and flow cytometry assay. Luciferase reporter assay was done to verify the targeted regulatory relationship of miR-98-5p with BMP-2. In comparison with NFH cases, DFH patients owned high levels of serum miR-98-5p and low concentration of BMP-2, and the levels of the two indexes are significantly negatively correlated. Both miR-98-5p and BMP-2 had the ability to predict DFH, while their combined diagnostic value is the highest. BMP-2 was demonstrated to be the target gene of miR-98-5p. Overexpression of BMP-2 reversed the role of miR-98-5p in MC3T3-E1 cell proliferation, apoptosis and differentiation. Increased miR-98-5p and decreased BMP-2 serve as potential biomarkers for the diagnosis of DFH. MiR-98-5p overexpression inhibits osteoblast proliferation and differentiation via targeting BMP-2.

[Role of macrophages in heart failure and traditional Chinese medicine intervention].

As the disease with high morbidity and mortality in the world, heart failure affects the development of human society. Due to its complicated pathology and limited treatment options, it is urgent to discover new disease targets and develop new treatment strategies. As innate immune cells accompanied by the evolution of heart failure, macrophages play an important role in cardiac homeostasis and stress. In recent years, the role of macrophages in the heart has attracted more and more attention as a potential target for heart failure intervention, and the research on cardiac macrophages has made important progress. Traditional Chinese medicine(TCM) has significant effects on regulating inflammatory response, treating heart failure, and maintaining homeostasis. In this article, researches on the functions of cardiac macrophages and application of TCM were reviewed from the source and classification of cardiac macrophages and the relationship of macrophages and cardiac inflammation, myocardial fibrosis, cardiac angiogenesis, and cardiac electrical conduction, which provided a basis for further basic research and clinical applications.

[Mechanism of Jiming Powder in ameliorating heart failure with preserved ejection fraction based on metabolomics].

In this study, untargeted metabolomics was conducted using the liquid chromatography-tandem mass spectrometry(LC-MS/MS) technique to analyze the potential biomarkers in the plasma of mice with heart failure with preserved ejection fraction(HFpEF) induced by a high-fat diet(HFD) and nitric oxide synthase inhibitor(Nω-nitro-L-arginine methyl ester hydrochloride, L-NAME) and explore the pharmacological effects and mechanism of Jiming Powder in improving HFpEF. Male C57BL/6N mice aged eight weeks were randomly assigned to a control group, a model group, an empagliflozin(10 mg·kg~(-1)·d~(-1)) group, and high-and low-dose Jiming Powder(14.3 and 7.15 g·kg~(-1)·d~(-1)) groups. Mice in the control group were fed on a low-fat diet, and mice in the model group and groups with drug intervention were fed on a high-fat diet. All mice had free access to water, with water in the model group and Jiming Powder groups being supplemented with L-NAME(0.5 g·L~(-1)). Drugs were administered on the first day of modeling, and 15 weeks later, blood pressure and cardiac function of the mice in each group were measured. Heart tissues were collected for hematoxylin-eosin(HE) staining to observe pathological changes and Masson's staining to observe myocardial collagen deposition. Untargeted metabolomics analysis was performed on the plasma collected from mice in each group, and metabolic pathway analysis was conducted using MetaboAnalyst 5.0. The results showed that the blood pressure was significantly lower and the myocardial concentric hypertrophy and left ventricular diastolic dysfunction were significantly improved in both the high-dose and low-dose Jiming Powder groups as compared with those in the model group. HE and Masson staining showed that both high-dose and low-dose Jiming Powder significantly alleviated myocardial fibrosis. In the metabolomics experiment, 23 potential biomarkers were identified and eight strongly correlated metabolic pathways were enriched, including linoleic acid metabolism, histidine metabolism, alpha-linolenic acid metabolism, glycerophospholipid metabolism, purine metabolism, porphyrin and chlorophyll metabolism, arachidonic acid metabolism, and pyrimidine metabolism. The study confirmed the pharmacological effects of Jiming Powder in lowering blood pressure and ameliorating HFpEF and revealed the mechanism of Jiming Powder using the metabolomics technique, providing experimental evidence for the clinical application of Jiming Powder in treating HFpEF and a new perspective for advancing and developing TCM therapy for HFpEF.

[Effect and mechanism of Jiming Powder on myocardial fibrosis in mice with myocardial infarction].

Jiming Powder is a traditional ancient prescription with good therapeutic effect in the treatment of heart failure, but its mechanism lacks further exploration. In this study, a mouse model of coronary artery ligation was used to evaluate the effect and mechanism of Jiming Powder on myocardial fibrosis in mice with myocardial infarction. The study constructed a mouse model of heart failure after myocardial infarction using the method of left anterior descending coronary artery ligation. The efficacy of Jiming Powder was evaluated from multiple angles, including ultrasound imaging, hematoxylin-eosin(HE) staining, Masson staining, Sirius Red staining, and serum myocardial enzyme spectrum detection. Western blot analysis was performed to detect key proteins involved in ventricular remodeling, including transforming growth factor-ß1(TGF-ß1), α-smooth muscle actin(α-SMA), wingless-type MMTV integration site family member 3a(Wnt3a), ß-catenin, matrix metallopeptidase 2(MMP2), matrix metallopeptidase 3(MMP3), TIMP metallopeptidase inhibitor 1(TIMP1), and TIMP metallopeptidase inhibitor 2(TIMP2). The results showed that compared with the model group, the high and low-dose Jiming Powder significantly reduced the left ventricular internal diameter in systole(LVID;s) and diastole(LVID;d), increased the left ventricular ejection fraction(LVEF) and left ventricular fractional shortening(LVFS), effectively improved cardiac function in mice after myocardial infarction, and effectively reduced the levels of myocardial injury markers such as creatine kinase(CK), creatine kinase isoenzyme(CK-MB), and lactic dehydrogenase(LDH), thus protecting ischemic myocardium. HE staining showed that Jiming Powder could attenuate myocardial inflammatory cell infiltration after myocardial infarction. Masson and Sirius Red staining demonstrated that Jiming Powder effectively inhibited myocardial fibrosis, reduced the collagen Ⅰ/Ⅲ ratio in myocardial tissues, and improved collagen remodeling after myocardial infarction. Western blot results showed that Jiming Powder reduced the expression of TGF-ß1, α-SMA, Wnt3a, and ß-catenin, decreased the levels of MMP2, MMP3, and TIMP2, and increased the level of TIMP1, suggesting its role in inhibiting cardiac fibroblast transformation, reducing extracellular matrix metabolism in myocardial cells, and lowering collagen Ⅰ and α-SMA content, thus exerting an anti-myocardial fibrosis effect after myocardial infarction. This study revealed the role of Jiming Powder in improving ventricular remodeling and treating myocardial infarction, laying the foundation for further research on the pharmacological effect of Jiming Powder.

[Common anti-inflammatory effects of heat-clearing and toxin-removing Chinese medicines on diverse cardiovascular diseases].

Cardiovascular diseases seriously affect human health and their prevalence continues to increase with the aging of the population. The integrated therapy of traditional Chinese medicine(TCM) and western medicine for cardiovascular diseases has achieved certain results, but it is still faced with new challenges. Studies have shown that inflammation plays an important role in the development of cardiovascular diseases and some of these mechanisms have common features. For example, in cardiovascular diseases, C-C motif chemokine receptor 2(CCR2)-expressing macrophages increase and promote inflammation, and excessive activation of NOD-like receptor protein 3(NLRP3) inflammasome leads to the elevation of inflammatory factors. There is also new understanding of the pathogenesis and treatment of cardiovascular diseases in TCM. The heat-toxicity theory in cardiovascular diseases and the therapeutic principle of clearing heat and removing toxin have attracted attention. The clinical and pharmacological studies on the treatment of cardiovascular diseases such as Huanglian Jiedu Decoction and Simiao Yong'an Decoction are also gradually increasing. The present study analyzed the common features of the inflammatory response mechanisms in diverse cardiovascular diseases and discussed the significance of the prevention and treatment of diverse cardiovascular diseases by the treatment method of clearing heat and removing toxin to regulate inflammation, which is expected to provide new ideas and references for clinical treatment and drug research on cardiovascular diseases with the same treatment method for different diseases.

circMTDH.4/miR-630/AEG-1 axis participates in the regulation of proliferation, migration, invasion, chemoresistance, and radioresistance of NSCLC.

Astrocyte elevated gene-1 (AEG-1) plays a critical role in the development, progression, and metastasis of a variety of cancers, including non-small-cell lung cancer (NSCLC). The objective of the current study is to unravel the upstream signaling of AEG-1. A cohort of 28 NSCLC tissues and 30 normal tissues were collected. Quantitative reverse transcription-polymerase chain reaction and Western blotting were used to examine AEG-1, migration, and invasion related markers in NSCLC cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay coupled with colony formation assay were conducted to monitor cell growth. Transwell assay was performed to determine cell migration and invasion. Apoptotic cells were detected by costaining with Annexin-V-fluorescein isothiocyanate and propidium iodide. Immunofluorescent staining was used to observe the levels of migration and invasion related markers. Xenograft models were used to investigate tumor formation in vivo. Dual-luciferase reporter assay and RNA immunoprecipitation were carried out to determine the interaction between circMTDH.4 and miR-630, as well as the associated between miR-630 and AEG-1. AEG-1 was highly expressed in NSCLC tissues and cell lines. Silencing of AEG-1 inhibited cell proliferation, migration, invasion, and chemoresistance/radioresistance in NCI-H1650 and A549 cells. circMTDH.4 regulated AEG-1 expression via sponging miR-630. Knockdown of circMTDH.4 and/or overexpression of miR-630 inhibited chemoresistance and radioresistance in NSCLC cells, whereas overexpression of AEG-1 or knockdown of miR-630 exerted rescue effects. circMTDH.4/miR-630/AEG-1 axis is responsible for chemoresistance and radioresistance in NSCLC cells.

Quantitative Proteomics and Cytology of Rice Pollen Sterol-Rich Membrane Domains Reveals Pre-established Cell Polarity Cues in Mature Pollen.

Cell polarity is essential for generating diverse cell functions. The underlying mechanisms of how a cell establishes, maintains, and changes its polarity are poorly understood. Recently, sterol-rich membrane microdomains are found to be associated with these processes. However, both its exact characteristics and importance are still unclear. Here we show microdomains change dynamically in developing and germinating rice pollen with selective enrichment in the aperture and the tip of newly born pollen tubes by use of the sterol-specific probe filipin. Using the sterol extraction sensitivities of microdomain proteins and quantitative proteomics, we identified 237 microdomain-associated proteins from 934 identified pollen detergent resistant membrane proteins. This proteome includes almost all of the known key regulators comprising the polar growth network, and it shows more similarity to front-back polarized HeLa cells than nonpolarized Arabidopsis suspension cells. We immunolocalize flotilin-like protein, a representative of these sterol-dependent proteins and directly visualize microdomains in pollen. These results indicate the presence of microdomains in pollen and pre-established cell polarity around the aperture during pollen maturation. Our findings reveal an atlas of the microdomain-associated proteome in pollen. This work provides useful resources and knowledge needed to further dissect the mechanisms for the establishment and maintenance of cell polarity.

Mass imaging of ketamine in a single scalp hair by MALDI-FTMS.

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) coupled with mass spectrometry imaging (MSI) is a rapidly emerging technology that produces distribution maps of small pharmaceutical molecules in situ in tissue sections. Segmental hair analysis provides useful information regarding the state and history of drug use. A preliminary MALDI-Fourier transform ion cyclotron resonance (FTICR)-MSI method was developed for direct identification and imaging of ketamine in hair samples. After decontamination, the scalp hair samples from ketamine users were scraped gently and were fixed onto a stainless steel MALDI plate using double-sided adhesive tape. A Bruker 9.4 T solariX FTICR mass spectrometer with continuous accumulation of selected ions function was used in the positive ion mode. Four single hairs from the same drug abuser were analyzed. Three of four single hairs demonstrated ketamine spatial distribution, while only traces of ketamine were identified in the other one. The platform could provide detection power of ketamine down to the 7.7 ng/mg level in hair. MALDI-FTICR-MSI demonstrated the drug distribution over the whole hair length with higher spatial resolution compared with the traditional LC-MS/MS method after scissor cutting. Greater caution is needed in the interpretation of a single hair result because of the considerable variations in the growth rate and sample collection.

Proteomic analysis of perfluorooctane sulfonate-induced apoptosis in human hepatic cells using the iTRAQ technique.

Perfluorooctane sulfonate (PFOS) is one of the most commonly used perfluorinated compounds, whose environmental exposure has been associated with a number of adverse health outcomes. However, the molecular mechanisms involved in PFOS toxicity are still not well elucidated. In the present study, we applied iTRAQ labeling quantitative proteomic technology to investigate the differential protein expression profiles of non-tumor human hepatic cells (L-02) exposed to PFOS. A total of 18 proteins were differentially expressed in a dose-dependent manner in PFOS-treated cells versus the control. Among these, 11 proteins were up-regulated and 7 were down-regulated. Gene ontology analysis indicated that PFOS would exert toxic effects on L-02 cells by affecting multiple biological processes, including protein biosynthesis and degradation, mRNA processing and splicing, transcription, signal transduction and transport. Furthermore, the proteomic results especially proposed that the inhibition of HNRNPC, HUWE1 and UBQLN1, as well as the induction of PAF1 is involved in the activation of the p53 and c-myc signaling pathways, which then trigger the apoptotic process in L-02 cells exposed to PFOS. Overall, these data will aid our understanding of the mechanisms responsible for PFOS-mediated hepatotoxicity, and develop useful biomarkers for monitoring and evaluating PFOS contamination in the environment.

Study on macro-meso mechanical properties of cemented tailings backfill with high fly ash content.

The use of cement and fly ash (FA) to prepare cemented tailings backfill (CTB) lowers backfill mining costs while also reducing pollution caused by the accumulation of waste materials like tailings and FA, making it a green backfill mining process. While adding FA to CTB may reduce costs, too much FA might weaken CTB's strength property. Mechanical tests were used to explore the effects of FA content and curing time on the uniaxial compressive strength (UCS) and deformation modulus of CTB in this research. The effects of FA content on failure modes, strain energy, and crack evolution of CTB were studied using a numerical model that considered FA content and particle contact mode. The influence mechanism of different FA contents on CTB was also revealed at the microscopic level. The results demonstrate that the UCS of CTB has a quadratic polynomial and a linear relationship with FA content and curing time respectively, and that the elasticity modulus and secant modulus of CTB increase and then decrease with FA content under different curing times. The peak strain energy of CTB increases and subsequently declines with the FA content, and crack propagation inside CTB may be limited by regulating the FA content. A reasonable content of FA can optimize the size and distribution of CTB microscopic defects, enabling them to exhibit superior strength property. This study systematically explores the mechanism of different FA contents on the strength property of CTB from a macro-micro perspective, providing an essential reference value for improving the recycling of FA and waste residues such as tailings.

Effect of calcium formate as an accelerator on dilatancy deformation, strength and microstructure of cemented tailings backfill.

Recycling mining wastes to produce cemented tailings backfill (CTB) is the optimal approach to eliminate the environmental pollution caused by their accumulation. However, its low strength limits its application. Using calcium formate (CF) as an accelerator for improving its mechanical properties is of great significance to promote sustainable development. The effects of CF dosage and curing time on dilatancy deformation, compressive strength and microstructure of CTB were investigated through mechanical compression, scanning electron microscope (SEM) and energy dispersive spectrometer (EDS) tests. The strengthening and deterioration mechanisms of CF dosage on CTB were revealed, and its engineering practicability was systematically evaluated. The results show that the variation of volumetric strain in the dilatancy deformation stage firstly increase and then decrease with the increases of CF dosage and curing time. The relationship between CF dosage and compressive strength can be characterized by quadratic polynomial, and the optimal CF dosage characterizing the superior mechanical property of CTB is between 1.60 and 1.84. The supplement of CF reduces the size and distribution of microcracks and micropores, thereby optimizing the microstructure of CTB. Nevertheless, the excessive dosages of CF deteriorate the microstructure of CTB and produce serious defects, which cannot be effectively filled by hydration products, thus weakening the strength property of CTB. This study provides an effective accelerator for improving the mechanical properties of CTB, which is of great significance to promote the recycling of tailings.

Influence of Microstructure on Dynamic Mechanical Behavior and Damage Evolution of Frozen-Thawed Sandstone Using Computed Tomography.

Frost-induced microstructure degradation of rocks is one of the main reasons for the changes in their dynamic mechanical behavior in cold environments. To this end, computed tomography (CT) was performed to quantify the changes in the microstructure of yellow sandstone after freeze-thaw (F-T) action. On this basis, the influence of the microscopic parameters on the dynamic mechanical behavior was studied. The results showed that the strain rate enhanced the dynamic mechanical properties, but the F-T-induced decrease in strength and elastic modulus increased with increasing strain rate. After 40 F-T cycles, the dynamic strength of the samples increased by 41% to 75.6 MPa when the strain rate was increased from 75 to 115 s-1, which is 2.5 times the static strength. Moreover, the dynamic strength and elastic modulus of the sample were linearly and negatively correlated with the fractal dimension and porosity, with the largest decrease rate at 115 s-1, indicating that the microscopic parameters have a crucial influence on dynamic mechanical behavior. When the fractal dimension was increased from 2.56 to 2.67, the dynamic peak strength of the samples under the three impact loads decreased by 43.7 MPa (75 s), 61.8 MPa (95 s), and 71.4 MPa (115 s), respectively. In addition, a damage evolution model under F-T and impact loading was developed considering porosity variation. It was found that the damage development in the sample was highly related to the strain rate and F-T damage. As the strain rate increases, the strain required for damage development gradually decreases with a lower increase rate. In contrast, the strain required for damage development in the sample increases with increasing F-T damage. The research results can be a reference for constructing and maintaining rock structures in cold regions.

Mechanical property and microstructure of cemented tailings backfill containing fly ash activated by calcium formate.

Cemented tailings backfill (CTB) is the most economical and environmental method to recycle tailings and fly ash (FA) for filling mining, but the high content of FA will weaken its strength property. This paper aims to use calcium formate (CF) as an activator to stimulate the activity of FA, thereby enhancing the mechanical property of CTB. The influence of FA and CF content on the stress-strain behavior, dilatancy deformation, and compressive strength of CTB was investigated using uniaxial compression test and scanning electron microscope. The coupling effect mechanism of FA and CF content on the compressive strength of CTB was revealed. The results show that increasing the content of FA and CF can enhance the bearing capacity of CTB during the dilatancy deformation stage, but the excessive content of FA and CF will lead to the attenuation of peak stress. The relations between FA content, CF content, and the compressive strength of CTB can be characterized by quadratic polynomial. Adding CF can stimulate the activity of insoluble FA, increasing the utilization of FA in CTB and producing rich hydration products to fill the internal defects of CTB. The microstructure of CTB is effectively improved by adding CF, including the size and distribution of microcracks and micropores, so that the strength property of CTB is optimized. However, too much CF will make the microstructure of CTB loose and porous, resulting in more microcracks and micropores. Microcracks propagate and connect with micropores to form defects, which deteriorate the microstructure of CTB, thus weakening the strength parameters of CTB. This study provides a method to increase the utilization of FA in CTB, which is of great significance for strengthening the mechanical properties of CTB and improving engineering economic benefits.

Coupled effects of fly ash and calcium formate on strength development of cemented tailings backfill.

Cemented tailings backfill (CTB) is widely adopted to ensure the safety of underground goafs and mitigate environmental risks. Fly ash (FA) and calcium formate (CF) are common industrial by-products that improve the mechanical performance of CTB. How the coupling of the two components affects the strength development is not yet well-understood. Neural network modelling was conducted to predict the strength development, including the static indicator of uniaxial compressive strength (UCS) and the dynamic indicator of ultrasonic pulse velocity (UPV). Sobol' sensitivity analysis was carried out to reveal the contributions of FA, CF and curing time to CTB strength. SEM microstructure investigation on CTB samples was implemented to reveal the mechanism of strength development and justify the predictions by neural network modelling and sensitivity analysis. Results show that the combination of FA content, CF content and curing time can be used to predict both UCS and UPV while providing adequate accuracy. The maximum of UCS of 6.1215 MPa is achieved at (FA content, CF content, curing time) = (13.78 w%, 3.76 w%, 28 days), and the maximum of UPV of 2.9887 km/s is arrived at (FA content, CF content, curing time) = (11.67 w%, 3.08 w%, 10 days). It is also implicated that prediction of UCS using UPV alone, although common in field application is not recommended. However, UPV measurement, in combination with the information of FA dosage, CF dosage and curing time, could be used to improve UCS prediction. The rank of variable significance for UCS is curing time > FA content > CF content, and for UPV is FA content > curing time > CF content; variable interaction is strongest for FA with CF for UCS development, and for FA with curing time for UPV evolution. Influence of FA on CTB strength development is due to improved polymerisation and consumption of Ca(OH)2. Influence of CF on strength development is a result of accelerated hydration and increased combined-water content in calcium silicate hydrate (CSH). Effect of curing time is attributed to the evolution of CSH product and pore-water content during cement hydration.

A proteomics analysis of rat liver membrane skeletons: the investigation of actin- and cytokeratin-based protein components.

Membrane skeletons, which are defined for their resistance to Triton extraction of cell membrane, play a pivotal role in cell shape and signal transduction. In the present work, we applied a complementary proteomics strategy: 2-DE combined with MALDI-TOF MS and 1D-PAGE coupled with LC-ESI-FTICR MS to analyze a membrane skeleton fraction isolated from Sprague-Dawley (SD) rat livers. We report confident identification of 104 proteins (39 membrane skeleton proteins) using 2-DE and MALDI-TOF MS approach and 402 proteins (87 membrane skeleton proteins) using 1D-PAGE LC-MS/MS analysis. In total, 100 membrane skeleton proteins were identified using the two complementary proteomics means. To the best of our knowledge, this is the largest data set of membrane skeleton proteins to date. Noteworthily, almost all of these membrane skeleton proteins were associated with actin or cytokeratin, and more than half of them were involved in various cell junctions. Our results offer insights into the protein components of the actin- and cytokeratin-based membrane skeletons in rat livers, which would improve our understanding of their biological roles.

Enrichment of serum low-molecular-weight proteins using C18 absorbent under urea/dithiothreitol denatured environment.

Serum low-molecular-weight proteins (LMWPs, molecular weight<30kDa) are closely related to the body physiological and pathological situations, whereas many difficulties are encountered when enriching and fractionating them. Using C(18) absorbent (100 A) enrichment and fractionation under urea/dithiothreitol (DTT) denatured environment followed by 60% acetonitrile (ACN) elution, serum LMWPs could be enriched more than 100-fold and were evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional gel electrophoresis (2-DE), and isotope-coded affinity tag (ICAT) labeling quantification. Proteins existing in human serum at low nanograms/milliliter (ng/ml) levels, such as myeloid-related proteins (MRPs), could be identified directly from 2-DE coupled with matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF MS) and LTQ-Orbitrap MS. Sixteen proteins were confidentially identified and quantified using ICAT labeling and liquid chromatography-tandem mass spectrometry (LC-MS/MS). By virtue of its easy operation and high reproducibility to process large quantity complex serum samples, this method has potential uses in enriching LMWPs either in serum or in cell and tissue samples.

Comparative studies of early liver dysfunction in senescence-accelerated mouse using mitochondrial proteomics approaches.

The liver is a complex and unique organ responsible for a breadth of functions crucial to sustaining life, especially for various metabolic processes in its mitochondria. Senescence-accelerated mouse prone/8 (SAMP8), a widely used aging model, exhibits an oxidative stress-induced aging phenotype and severe mitochondria-related liver pathology that are not seen in senescence-accelerated mouse resistant/1 (SAMR1). Here we used both two-dimensional electrophoresis- and ICAT-based mitochondrial proteomics analysis to view the liver mitochondrial protein alterations between SAMP8 and SAMR1. Compared with SAMR1, decreased expression and activity of mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase were detected in SAMP8 at 6 months old (SAMP8-6m). As the key enzyme of ketogenesis, 3-hydroxy-3-methylglutaryl-CoA synthase is well known to be transcriptionally regulated by peroxisome proliferator-activated receptor alpha, which was also expressed at lower levels in SAMP8-6m livers. In addition, down-regulation of two peroxisome proliferator-activated receptor alpha target gene products (acyl-CoA oxidase and enoyl-CoA hydratase), elevation of triglyceride, and reduction of acetyl-CoA were observed, indicating abnormal fatty acid metabolism in SAMP8-6m livers. In addition eight proteins (NDUAA, NDUBA, NDUB7, NDUS1, NDUS3, NDUV1, ETFA, and UCRI) of mitochondrial complexes were down-regulated in SAMP8-6m, resulting in mitochondria-related liver dysfunction characterized by enhanced oxidative stress-induced molecular damage (lipid peroxide and oxidized protein) and depressed energy production (ATP). Glutamine synthetase and ornithine aminotransferase involved in glutamine synthesis were up-regulated in SAMP8 livers at both 1 and 6 months old that may be related to the accumulation of glutamate and glutamine. Our work provided useful clues to understanding the molecular mechanism underlying liver dysfunction in senescence-accelerated mouse.

O,O'-Diethyl {(Z)-[(2-chloro-phen-yl)(cyano)methyl-ene]amino-oxy}thio-phospho-nate.

The title mol-ecule, C(12)H(14)ClN(2)O(3)PS, has a cis configuration with respect to the C=N bond. Inter-molecular C-H⋯O inter-actions inter-connect the mol-ecules into chains along the c axis. The chains are further connected into a two-dimensional network parallel to the bc plane by weak π-π inter-actions between adjacent aromatic rings (centroid-centroid distance = 3.772Å).

The Promoting Effect of Radiation on Glucose Metabolism in Breast Cancer Cells under the Treatment of Cobalt Chloride.

We aimed to investigate the influence of radiation on hypoxia-treated breast cancers cells and its underlying mechanism. We mimicked the hypoxic response in MCF-7 cells by the treatment of CoCl2. Meanwhile, hypoxic MCF-7 cells induced by CoCl2 or untreated MCF-7 cells were treated with or without radiation, and then treated with or without hypoxia inducible factors-1α (HIF-1α) inhibitor. Subsequently, glucose update and lactate release rate were determined by commercial kits, as well as the expressions of HIF-1α and the glucose metabolic pathway related genes, including fructose biphoshatase 1 (FBP1), glucose transporter 1 (GLUT1), lactate dehydrogenase A (LDHA), hexokinase 2 (HK2), and isocitrate dehydrogenase 2 (IDH20) were detected by western blotting and/or RT-PCR. The results showed that glucose uptake rate and lactate release rate were increased in cells under hypoxia and/or radiation condition compared with untreated cells (p < 0.05), while the addition of HIF-1α inhibitor decreased these rates in hypoxia + radiation treated cells (p < 0.05). In addition, compared with untreated cells, the mRNA and protein levels of HIF-1α were significantly increased under hypoxia and radiation condition (p < 0.05), while which decreased after the addition of HIF-1α inhibitor (p < 0.05). Similar content changing trends (all p < 0.05) were observed in FBP1, IDH2, GLUT1, and LDHA but not HK2. In conclusion, the combination of radiation and hypoxia could promote the glucose metabolism. Furthermore, HIF-1α might inhibit the promoting effect of radiation on glycolysis in hypoxic MCF-7 cells by regulating the glucose metabolic pathway.

[Identification of biomarkers in serum of early rheumatoid arthritis by proteomic methods].

OBJECTIVE: To discover the biomarkers of early rheumatoid arthritis (RA) patients and healthy person were analyzed by proteomic methods to discover serum biomarkers. METHODS: Samples of peripheral blood were collected form 10 newly diagnosed active RA patients, 5 males and 5 females, aged 54.3 +/- 12.78, with a disease course of 4.08 +/- 1.9 months, and 10 age and sex matched healthy persons. The samples were divided into 10 groups containing 1 sample of patient serum and 1 sample of healthy serum to undergo. High-molecular-weight protein (HMWP) was enriched by depletion of albumin with HiTrap Blue column and depletion of immunoglobulin G with HiTrap rProtein A column. Low-molecular-weight protein (LMWP) was enriched by C18 absorbent binding and elution. After comparative proteomic analysis, the different protein spots were identified using matrix-assisted laser desorption ionization-time of flight-ionization-mass spectrometry. (MALDI-TOF-MS). RESULTS: Marrow-related protein (MRP) 14, a protein of the S100 family was 100% positive in the RA patient sera and 100% negative in the sera of the healthy controls. MRP8, another protein of the S100 family, was 100% positive in the RA patient sera and 50% positive in the sera of the healthy controls. Ubiquitin was 90% positive in the RA patient sera and 10% positive in the sera of the healthy controls. The levels of apolipoprotein A-I (ApoA-I), serum amyloid A1 and A2 (SAA1/SAA2), and transthyretin of the RA patients were all significantly higher than those of the healthy controls. CONCLUSION: MRP14/MRP8, SAA1/SAA2 and ubiquitin may play important roles in development of RA and their determination may benefit early diagnosis, evaluation of disease activity and investigation of new therapy targets.