Central Nervous System ; physiopathology ; Facial Nerve Injuries ; physiopathology ; Humans ; Neuronal Plasticity ; Synapses ; physiology

This study investigates whether kappa-opioid receptor and ORL1 receptor may interact to form a heterodimer. In immunofluorescence and co-immunoprecipitation experiments, differentially epitope-tagged receptors, colocalization and heterodimerization of kappa-opioid receptor and ORL1 receptor were used and examined in primary culturing rat neurons, Chinese hamster ovary (CHO) or human embryonic kidney 293 (HEK293) cells. The results show that fluorescence of both kappa-opioid receptor and ORL1 receptor were overlapping in primary culturing hippocampal and cortical neurons. Similarly in co-expressing CHO or HEK293 cells, HA-KOR and Myc-ORL1 were almost exclusively confined to the membranes, revealing extensive colocalization. When Flag-KOR and Myc-ORL1 were co-expressing in CHO cells, heterodimerization was identified to have the ability to co-immunoprecipitate ORL1-receptors with kappa-opioid receptor and vice versa. In the current study, further evidence was provided for the direct interaction of two subtypes of opioid receptors, kappa-opioid receptor and ORL1-receptor, to form the heterodimerization. The finding represents the novel pharmacological mechanism for modulation of opioid receptor function as well as diversity of G protein-coupled receptors.

A rapid method was developed for the determination of 9 organophosphorous flame retardants ( OPFRs) in textiles by ultra high performance liquid chromatography tandem electrospray ionization mass spectrometry ( UHPLC-MS/MS ) . Analytes in textile samples were extracted in a closed bottle at 50℃ with methanol by ultrasonic extraction for 30 min. And then, the extracted solution was injected after filtration and determined by UHPLC-MS/MS. Target analytes were separated by C18 column and methanol/water containing 5. 0 mmol/L NH4 COOH. The positive electrospray ionization and multiple reaction monitoring mode ( MRM) was utilized. The matrix effect (ME) could be compensated by adjusting the concentration of NH4COOH and the composition of the mobile phase after careful investigation with blank samples. An external standard calibration method was chosen to the quantitative analysis. Under the optimized conditions, method precision represent with relative standard deviations (RSDs) was 5. 2% (TOCP) -12. 5% (TRIS) for cotton, 6. 4%(TBEP)-13. 8% (TRIS) for polyester, and 6. 9% (TDCP) -14. 5% (TEPA) for wool samples. The recoveries for standard addition were in the range of 52 . 5% ( TEPA ) -116 . 4% ( TPP ) . The limits of quantification (LOQ, S/N=10) were between 1. 0 (TBP) and 10. 0 (TRIS) μg/kg.

Akt1 is a serine-threonine protein kinase that has been implicated in the control of cellular metabolism,survival and growth.Elevated expression of Akt1 has been noted in a significant percentage of human tumors,promoting cellular metastasis.Conversely,some studies have revealed hyperactivated Akt1 inhibited the invasiveness and metastasis of breast cancer cells.To clarify the definite effect of Akt1 on tumorigenesis and development,Akt1 was silenced by RNAi in the highly metastatic murine breast cancer 4T1 cells.Akt1 silencing didn't affect the proliferation of breast cancer cells in MTT assay,while reduced the migration in Transwell assay.Consistent with the above results,Akt1 silencing didn't change the primary tumor weight,but significantly suppressed lung metastasis of 4T1 cells.These observations indicated Akt1 plays an important role in murine breast cancer metastasis,and suggested that Akt1 might be a therapeutic target for breast cancer metastasis.

Adult ; Carcinogens ; Epoxy Compounds ; poisoning ; Female ; Humans ; Lymphocytes ; drug effects ; metabolism ; Male ; Multivariate Analysis ; Mutation ; drug effects ; Occupational Exposure ; analysis ; Regression Analysis ; Sister Chromatid Exchange ; drug effects

AIM:TodetecthemoglobinA1c(HbA1c)andparametersofbloodglucosefluctuationinChinesenewlydiag-nosed type 2 diabetes mellitus (T2DM) patients, and further to specify the factors that were related to mean blood glucose (MBG) in this population.METHODS:Newly diagnosed T2DM patients (n=90) from 4 hospitals in Guangdong province were enrolled, and subjected to 3 d continuous glucose monitoring (CGM) after testing for HbA1c and other laboratory tests.Blood glucose data collected during CGM were used to calculate MBG and parameters of blood glucose fluctuation.RESULTS: Correlation analysis revealed that MBG was significantly related to all parameters of blood glucose fluctuation, HbA1c, fast plasma glucose ( FPG) and 2 h postprandial glucose (P<0.01), but not to sex, age or blood lipid profile.Further analysis utilizing step-wise general linear model showed that HbA1c, absolute means of daily difference ( MODD) , difference between maximal and minimal glucose ( DMMG) and FPG had the strongest relation to MBG.CONCLUSION: Factors affecting MBG of the newly diagnosed T2DMpatients in our country include HbA1c, FPG, DMMG and MODD, and thus it may be prone to misleading results that only HbA1c is applied to estimate MBG in this population.

To confirm the etiology of a dead case for a 6 year-old female Ailurus fulgens,one strain of the predominant bacteria from pathologic tissues(heart,liver,spleen,lung and other samples) of the dead Ailurus fulgens were examined and isolated.The isolate was named R1 and no other bacteria were isolated.The bacterial etiological examination(morphological characteristics,biochemical characteristics and 16S rDNA gene detection)of R1 showed that it was identifed as K.peneumoniae.Artificial infection to mice about R1 was also conducted in this study.R1 had strong pathogenicity to mice and the LD50 is 6.5 × 104 CFU/mL.Moreover,the clinical and pathological features of the dead mice were consistent with that of the Ailurus fulgens.To find effective therapeutic drugs of curing other Ailurus fulgens,antibiotic sensitivity test of R1 was conducted,and the results revealed that R1 was highly sensitive to cefotaxime et al,moderately sensitive to amikacin and resistant to penicillin.These data showed that K.peneumoniae was bacterial pathogen leading to death of the Ailurus fulgens and it had strong resistance to penicillins,macrolides and virginiamycin and it had broad drug resistance spectrum.However,R1 is sensitive to cephalosporins and aminoglycoside antibiotics.

10.3969/j.issn.1007-3969.2013.04.00X

Toad venom is the Bufo bufo gargarizans or B. melanostictus after the ears of the gland secretion, used in the treatment of various cancers in recent years. Research shows that the main anti-tumor components in bufadienolide. Bufadienolide have free type structure and conjunct type structure. To identify and clarify the difference between bufogenin and bufotoxin contained in Bufonis Venenum, which was from B. bufo gargarizans, an UPLC-TQ-MS method has been established. UPLC-TQ-MS method was used to identify and quantify the major bufadienolides in Bufonis Venenum. UPLC-TQ-MS assay with positive ion mode was performed on a Waters ACQUITY UPLC BEH C, (2.1 mm x 100 mm, 1.7 µm) with the mobile phase consisting of 0. 1% aqueous formic and acidacetonitrile in gradient elution at a flow rate of 0.4 mL · min⁻¹ and the column temperature was set at 35 °C. By comparing their retention time and high resolution mass data of Bufonis Venenum extracts, 37 effective components were primarily identified by MS/MS analysis in positive ion mode. Twenty-six of them were free-type bufadienolides (bufogenin), 11 of them were conjugated bufadienolides. There were significant differences in the main composition between fresh and processed Bufonis Venenum. The study found that the chemical composition of toad venom through great changes after processing, conjunct type content is much less, free type content as well change.
Amphibian Venoms ; chemistry ; metabolism ; Animals ; Bufonidae ; classification ; metabolism ; Chromatography, High Pressure Liquid ; methods ; Molecular Structure ; Tandem Mass Spectrometry ; methods

AIMTo study the effects of prophylene glycol mannate sulfate (PGMS) on monocyte chemoattractant protein-1 (MCP-1) mRNA expression in hyperlipidemic rat aorta and to clarify the molecular mechanism of PGMS for the prevention of atherosclerosis.

METHODSPGMS (37.8 and 75.6 mg.kg-1.d-1, ig) or PGMS (37.8 and 75.6 mg.kg-1.d-1, ig) combined with diethyldithiocarbamate (DDC, an inhibitor of SOD, 200 mg.kg-1 every three days, i.p.) were given to hyperlipidemic rats for three weeks. The MDA content and SOD activity were determined after 12 h of starvation, and MCP-1 mRNA expression in aorta was detected by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThere was significant decrease (29.46% or 58.40)% of MCP-1 mRNA expression in aortic after the therapy. The SOD activity increased markedly and the MDA content decreased at the same time. After treatment with DDC, the SOD activity was inhibited and the MDA content increased, but with no significant effect on MCP-1 mRNA expression.

CONCLUSIONPGMS inhibited MCP-1 mRNA expression with no relation to its effect on decreasing MDA content.

Animals ; Aorta, Thoracic ; drug effects ; metabolism ; Chemokine CCL2 ; biosynthesis ; genetics ; Gene Expression ; drug effects ; Hyperlipidemias ; blood ; pathology ; Hypolipidemic Agents ; pharmacology ; Male ; Malondialdehyde ; blood ; metabolism ; Propylene Glycols ; pharmacology ; RNA, Messenger ; biosynthesis ; drug effects ; Random Allocation ; Rats ; Rats, Wistar ; Superoxide Dismutase ; blood ; metabolism