OBJECTIVETo study the effects and mechanism of apigenin on the expression of vascular endothelial growth factor (VEGF) in human breast cancer MDA-MB-231 cells.

METHODSMDA-MB-231 cells were used as the study object. MTT assay was used to detect the effect of apigenin on the cell viability. ELISA was used to determine the protein level of VEGF. RT-PCR was used to detect VEGF at mRNA level. A double luciferase system was used to measure the transcription activity of VEGF. pCEP4-HIF-1alpha was transferred to explore the reversing effect of HIF-1alpha on the inhibitory effect of apigenin on the transcription activity of VEGF. Western blotting was used to detect the time-dependent and dose-dependent effect of apigenin on HIF-lalpha, p-AKT, p-ERK, and p53 expression at protein level.

RESULTSApigenin had no visible inhibitory effect on cell viability. Apigenin reduced the secretion of VEGF, mRNA levels of VEGF and transcription activity of VEGF. Furthermore, the inhibitory effect of apigenin on the transcription activity of VEGF could be reversed by transferring pCEP4-HIF-1alpha into the cells. Additionally, apigenin inhibited the expression of HIF-1alpha and p-AKT, induced the expression of p53, but had no effect on the expression of p-ERK1/2.

CONCLUSIONApigenin can inhibit VEGF expression in breast cancer cells, and this effect may be achieved through decreasing the expression of HIF-1alpha.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apigenin ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; metabolism ; Transcriptional Activation ; drug effects ; Tumor Suppressor Protein p53 ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Objective To investigate the pathogenesis,diagnosis,treatment and prognosis of aort-oesophageal fistula(AEF).Methods Retropective analysis was performed on 6 patients presenting with AEF between January 2002 and December 2014,and relative literature was reviewed on its pathogenesis,di-agnosis,prognosis and treatment.Results Five men and 1 woman with a mean age of 49 (range,27-71 years)were recruited to the study.One case of AEF was caused by esophageal foreign body,2 cases were caused by aneurysm while the other 3 patients presented AEF after aortic surgery.All 6 patients showed he-matemesis,among whom 3 presented sentinel hemorrhage,1 presented exsanguination after sentinel hemor-rhage,2 presented sudden exsanguination.Among 4 patients with sentinel hemorrhage,2 accompanied with chest pain,1 with dysphagia and 1 with fever.Two patients had a history of hypertension.Diagnostic rate was nearly 100% by gastroscopy or CT/CTA.Four patients died from hemorrhagic shock and 2 patients re-covered from surgery.Conclusion AEF should be seriously considered for patients with a history of hyper-tension,aortic disease or esophageal foreign body presenting sentinel hemorrhage,chest pain,dysphagia,fa-tal exsanguination followed by symptom-free interval.Prompt examinations and aggressive surgery are of great significance for survival.

Objective To compare four methods of monitoring vascularization of tissue engineered bone in the rhesus so as to find our the best. Methods Twenty-five lower limbs of 13 rhesuses were used in this study to make models of tibial diaphyseal defect of 20mm which were to be fixed with an AO reconstruction plate of 7 holes. The monkeys were randomly divided into five groups according to defect filling materials: group A:?-tricalcium phosphate (?-TCP) and bone marrow stromal cells (BMSCs) and blood vessel bundles; group B:?-TCP and blood vessel bundles; group C:?-TCP and BMSCs; group D:?-TCP; group E: blank. Perfusion weighted MR imaging (PWMR), X-ray, radionuclide imaging and histological examinations were carried out at weeks 4, 8, 12 postop- eratively. The maximum slope rates of the single intensity-time curve (SS_(max)) and values of baseline (Sl_(?))were calculated at the same time points. Transmittances of the X-ray films were assessed. Ratios between isotope counts in region of interest (ROI) were calculated. Chinese ink perfusion and calculation of blood vessel areas were done for histological examinations, Results Compared with other groups, the SS_(max) in group A was the highest at weeks 4, 8, 12 postoperatively. In group A, the SS_(max) at week eight was significantly higher than that at week four (P= 0. 003), and the SS_(max) and transmittance of X-ray were negatively related at week 12 after operation (rs=-0. 892, P=0. 042), but the SS_(max) and blood vessel area were positively related (rs=0. 894, P=0.041)Conclusions PWMR can be a sensitive, quantitative, noninvasive and non-radiant method to monitor vascularization of tissue engineered bone, because SS_(max) of the single intensity-time curve of PWMR can reflect the most accurately the process of vascularization of tissue engineered bone.

Objective:To analyze the effects of espO gene knockout on the biological characteristics of enterhemorrhagic Escherichia coli (EHEC). Methods:Two-step methods mediated by the suicide plasmid pCVD442-Δ espO and plasmid pTrc99a were used to construct the espO gene-deleted strain (Δ espO) and the complemented mutant (CΔ espO), respectively. HeLa cells were infected with different EHEC strains to analyze the biological functions and lethal effects of espO gene during infection. Results:PCR, electrophoresis and gene sequencing showed that the Δ espO and CΔ espO mutants were successfully constructed. Compared with the wild-type strain, neither the Δ espO nor CΔ espO mutant showed significant difference in growth rate, indicating that the espO gene had no influence on the growth and replication of EHEC. Furthermore, EspO could activate the tumor necrosis factor receptor (TNF)-induced NF-κB signaling pathway, while the effector protein NleB could inhibit the process. EspO could not inhibit the death of HeLa cells induced by TNF or TNF-related apoptosis-inducing ligand (TRAIL) after EHEC infection. Conclusions:In this study, we successfully constructed the espO gene-deleted and complemented mutants of EHEC and preliminarily analyzed the interaction between espO gene and host cells and the effects of espO gene on cell apoptosis during infection, which provided reference for further research on the in vitro biochemical activity and in vivo pathogenic roles of EspO.

The effects of somatic and visceral nociceptive stimulation on neurons of dorsal commissural nucleus in sacral spinal cord were studied in pentobarbitone anesthetized cats using extracellular recording techniques. The results indicated that all neurons deriving input from Aδ fibers of the pelvic nerve were either nociceptive specific or wide dynamic range neurons, and could be activated by mechanical or intense electrical stimulation of the respective receptive fields including perianal region. The above results suggest that the Aδ afferents innervation of the pelvic organ is important in nociception.

To establish quality standards of Euonymus fortunei, and supply scientific evidence for the quality control of Euonymus fortunei. Empirical and microscopic identification methods were adopted to observe morphological and histological characters. The contents of water, total ash, acid-insoluble ash and alcohol-soluble extractive were analysed according to the methods of Chinese Pharmaco- poeia (2010). Dulcitol and reference herbs were used to identify materia medica of Euonymus fortunei by TLC method. The total flavonol glycosides contents were analysed by HPLC method, using quercetin and kaempferol as reference substances. Quercetin and kaempferol were separated on a C18 column (4.6 mm x 250 mm, 5 μm) with methanol-0.1% formic acid(51:49) as the mobile phase and detected at 366 nm. The flavonoid aglycones content was then multiplied by a conversion coefficient, and the result was the total flavonol glycosides content. The macroscopical identification, microscopic features and TLC methods were proper. The average contents of water, total ash, acid-insoluble ash, alcohol-soluble extractive and total flavonol glycosides were 8.76%, 6.48%, 0.31%, 17.48% and 0.211% , respectively. The quality standards established on the basis of the research results were suitable for the quality evaluation of Euonymus fortunei.
China ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Drugs, Chinese Herbal ; chemistry ; standards ; Euonymus ; anatomy & histology ; chemistry ; Mass Spectrometry ; Quality Control

Studies have found that metformin is not only the preferred drug for lowering blood sugar, but also shows lipid-lowering and weight-loss effects. The purpose of this study was to use a hyperlipidemia hamster model to investigate the lipid-lowering effect of metformin and its effect on important metabolic pathways in lipid metabolism disorders. Fifty golden hamsters were divided into a control group, a model group, metformin high- and low-dose groups, and a simvastatin group. A high-fat diet was fed for 1 week to create the model, and then drug was administered for 11 weeks with the high-fat diet. Serum was taken for measurement of blood lipid and blood glucose at 2, 6, and 9 weeks after administration, and at weeks 3, 5, and 9 feces and urine were collected for ¹H NMR metabolomics tests. After 11 weeks of intravenous injection of [U-¹³C₆] glucose, serum was collected for a ¹³C NMR metabolic flux test. The results showed that the administration of metformin can significantly reduce blood lipids and glucose levels and can significantly affect metabolic pathways such as sugar metabolism, lipid metabolism, ketone metabolism, amino acid metabolism, and intestinal flora metabolism. The results of the metabolic flux analysis showed that the high-fat diet reduced the metabolism of tricarboxylic acids by 37.48%. After administration of low and high doses of metformin the metabolism of tricarboxylic acid increased by 98.14% and 143.10%, respectively. After administration of simvastatin tricarboxylic acid metabolism increased by 33.18%. The results indicate that metformin has a significant effect on promoting energy metabolism. This study used a combination of metabolomics and metabolic flow to explore the effect of metformin on lipid metabolism disorders and quantifies changes in the key pathway of energy metabolism-the tricarboxylic acid cycle. This study provides useful information for the study of the efficacy and mechanism of metformin, as well as a practical technical method for the screening of lipid-lowering drugs based on a hamster model.

Lysostaphin is highly effective on eliminating methicillin resistant Staphylococcus aureus (MRSA). In order to achieve controlled release of lysostaphin, a biocompatible drug carrier is needed. Hydroxyapatite/chitosan (HA/CS) composites were chosen to carry lysostaphin and sample composites with different weight ratios of HA to CS, including 80/20, 70/30, 60/40, and 40/60, were prepared. Multiple analyses were performed to determine the structural and physicochemical properties of the composites, including scanning electron microscopy, X-ray diffraction and Fourier transform infrared spectroscopy. We immersed HA/CS composites loaded with 1 wt% lysostaphin to test in vitro release activity and cultured MC3T3-E1 cells to carry out biocompatibility test. The result of the release behavior of the composites revealed that the controlled release of lysostaphin from 60/40 HA/CS composites was the highest release rate of (87.4 ± 2.8)%, which lasted for 120 hours. In biocompatibility testing, MC3T3-E1 cells were able to proliferate on the surface of these composites, and the extract liquid from the composites could increase the growth of the cells. These results demonstrate the controlled release of lysostaphin from HA/CS composites and their biocompatibility, suggesting the potential application of these composites to bone injury and infection applications.
3T3 Cells ; Animals ; Biocompatible Materials ; Chitosan ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; chemistry ; Durapatite ; chemistry ; Lysostaphin ; pharmacology ; Materials Testing ; Methicillin-Resistant Staphylococcus aureus ; Mice ; Microscopy, Electron, Scanning ; X-Ray Diffraction

Objective To study the improved zygomatic reduction in the prevention of injury in the frontal branch of facial nerve.Methods Sixty-two patients with protrusion zygomatic complex,received treatment in our hospital from March 2008 to April 2010,were chosen in our study; 32 were given improved zygomatic reduction,and the other 30 were performed traditional zygomatic reduction.The clinical data of these patients,injury of the frontal branch of facial nerve and incidence of complication in these patients were compared between the 2 groups.Results No one suffered from injury of the facial nerve in the 32 patients given improved zygomatic reduction,but 3 suffered from nerve injury (mid injury in 2 and heavy injury in 1) in patients received traditional zygomatic reduction; 3 patients could not move the eyebow,2 suffered from sensory disability,and 1 had no wrinkle of forehead in patients received traditional zygomatic reduction.Patients given improved zygomatic reduction did not have the above complications.Conclusion Improved zygomatic reduction can effectively prevent injury in the frontal branch of facial nerve.

INTRODUCTIONCirculating insulin concentrations provide important information for the evaluation of insulin secretion and insulin resistance. Reference intervals are the most widely applied tool for the interpretation of clinical laboratory results. We carried out an analysis of the data available from the Fangchenggang Area Male Health and Examination Survey in order to derive a reference interval for fasting insulin specific to the Chinese population.

METHODSA total of 1,434 fasting serum insulin results were obtained from healthy nondiabetic adult men aged 20-69 years, after taking into consideration the inclusion and exclusion criteria. Serum insulin was measured using electrochemiluminescence immunoassays. Nonparametric statistical methods were used to calculate and analyse the data.

RESULTSThe reference interval for fasting serum insulin for Chinese adults was in the range 1.57-16.32 μU/mL (median 5.79 μU/mL). Significant correlations were found between fasting serum insulin and glucose and diastolic blood pressure (p < 0.001). Statistically significant differences were observed in insulin concentration with respect to age and body mass index (BMI; p < 0.001). Younger people had a higher fasting serum insulin concentration. Increased fasting serum insulin was also found to be associated with BMI.

CONCLUSIONWe established a reference interval for fasting serum insulin in healthy nondiabetic adult Chinese men that is lower than what was previously suggested. BMI and age (but not smoking, alcohol consumption or physical activity) were found to be important factors associated with fasting serum insulin. Our results will help improve the diagnostic interpretation of investigations for metabolic and cardiovascular disorders in a Chinese population.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Body Mass Index ; China ; Fasting ; blood ; Humans ; Insulin ; blood ; Insulin Resistance ; physiology ; Male ; Middle Aged ; Nomograms ; Reference Values ; Retrospective Studies ; Young Adult