Aim To investigate the effects of U50488H,a selective ?-opioid receptor agonist,on the blood pressure in the rats and explore their mechanisms.Methods Heart rate(HR),arterial blood pressure(ABP),left ventricular pressure(LVP),contractive function(+ dp/dt_(max)) and diastolic function(-dp/dt_(max)) were examined in rats.Physiological experimental technique was used to collect urine and to determine the volume of urine output;isolated artery perfusion technique was used to investigate the direct action of U50488H on abdominal aorta of rats.Results HR,ABP,LVP and ?dp/dt_(max) in rats were decreased with the administration of U50488H;the urine volume increased significantly with the administration of U50488H.U50488H induced a dose-dependent vasodilation in the aortic artery.These effects of U50488H were totally abolished by nor-BNI,a selective ?-opioid receptor antagonist.Conclusion Stimulation of ?-opioid receptor with U50488H depresses the blood pressure mainly by reducing the strength of cardiac muscle,enhancing the urine volume and relaxing the vessel.

The viability and life span of pollen were evaluated by TTC (2,3,5-triphenyl tetrazlium chloride) and the peroxidase solution, the stigma receptivity were estimated by benzidine-H2O2 method and the fruiting characteristics were investigated. The results showed that (1) Anoectochilus roxburghii and A. formosanus appeared the same up-and-down trend of the pollen viability, increased and then decreased. The storage temperature and storage time had significant impact on the pollen viability. With the extension of storage time, the pollen activity decreased. 4 degrees C refrigerator storage may be extended the pollen vitality. (2) The stigma had receptivity in 1st day and reached the highest level in the 4th day after blooming. A. roxburghii lost receptivity in the 8th day while A. formosanus lost receptivity in the 10th day after blooming. (3) The different pollination had significant impact on seed setting rate. The seed setting rate of artificial cross-pollination was higher than that of the artificial self-pollination. Collecting pollen in the 3rd day and carrying out artificial cross-pollination in the 4th day after blooming can significantly improve seed setting rate. The results provided technical assurance for A. roxburghii and A. formosanus breeding of new varieties and seed breeding.
Cell Survival ; China ; Flowers ; physiology ; Fruit ; growth & development ; Orchidaceae ; growth & development ; physiology ; Pollen ; growth & development ; Reproduction ; Temperature

This study is designed to obtain recombinant human acetylcholinesterase (rhAChE) and apply it in screening acetylcholinesterase inhibitors. The rhAChE was overexpressed in HEK293 cells transfected by plasmid of pCMV-AChE with the cationic liposome and rhAChE was found to be secreted into cell culture medium. AChE activity was assayed according to modified Ellman method to obtain kinetic parameters. IC so50 values for donepezil compounds of rhAChE were calculated to determine their activities of inhibition. The results showed that Km value was 151.9 micromol.L-1 donepezil inhibited rhAChE in a mixed competitive-noncompetitive way (Ki= 16.03 nmol.L-1, Ki = 18.36 nmol.L-1) and that most new compounds tested exhibited high activities of inhibition on rhAChE. The study suggests that rhAChE is available to be applied in screening AChE inhibitors in vitro.
Acetylcholinesterase ; genetics ; metabolism ; Cholinesterase Inhibitors ; analysis ; pharmacology ; HEK293 Cells ; Humans ; Indans ; analysis ; pharmacology ; Inhibitory Concentration 50 ; Kinetics ; Piperidines ; analysis ; pharmacology ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection

OBJECTIVE: To explore the relation between the behavior psychology analysis of partial mechanical injuries and the nature of death in high-falling cases, and provide reference, for such cases. METHODS: Of 311 death victims of high-falling injuries collected from 2008 to 2013, 205 cases were associated with partial mechanical injuries. The characteristics of injury formation, preliminary crime scene traces, fatal injury of high-falling, and text messages were all retrospectively analyzed. RESULTS: According to the investigation of preliminary crime scene traces, fatal injury of high-falling and text message, there were 86 suicide, 24 accident and 95 uncertainty in the 205 cases. According to the behavior psychology analysis of partial mechanical injuries, there were 80 suicide, 11 accident, and 4 homicide in the 95 uncertainty cases. CONCLUSION The partial mechanical injuries uncertainly caused by high-falling correlate with the manner of high-falling death. According to the behavior psychology analysis of the partial mechanical injuries in high-falling death cases, the presumption of high-falling death is usually accurate
Accidents/statistics & numerical data* ; Cause of Death ; Death ; Female ; Forensic Pathology ; Homicide/statistics & numerical data* ; Humans ; Male ; Retrospective Studies ; Suicide/statistics & numerical data* ; Uncertainty ; Wounds and Injuries/pathology*

AIMTo investigate the expression and distribution of the members of the transient receptor potential (TRP) channel members of TRP melastatin (TRPM) and TRP vanilloid (TRPV) subfamilies in rat prostatic tissue.

METHODSProstate tissue was obtained from male Sprague-Dawley rats. Reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time polymerase chain reaction (PCR) were used to check the expression of all TRPM and TRPV channel members with specific primers. Immunohistochemistry staining for TRPM8 and TRPV1 were also performed in rat tissues.

RESULTSTRPM2, TRPM3, TRPM4, TRPM6, TRPM7, TRPM8, TRPV2 and TRPV4 mRNA were detected in all rat prostatic tissues. Very weak signals for TRPM1, TRPV1 and TRPV3 were also detected. The mRNA of TRPM5, TRPV5 and TRPV6 were not detected in all RT-PCR experiments. Quantitative real-time RT-PCR showed that TRPM2, TRPM3, TRPM4, TRPM8, TRPV2 and TRPV4 were the most abundantly expressed TRPM and TRPV subtypes, respectively. Fluorescence immunohistochemistry indicated that TRPM8 and TRPV1 are highly expressed in both epithelial and smooth muscle cells.

CONCLUSIONOur results demonstrate that mRNA or protein for TRPM1, TRPM2, TRPM3, TRPM4, TRPM6, TRPM7, TRPM8, TRPV1, TRPV2, TRPV3 and TRPV4 exist in rat prostatic tissue. The data presented here assists in elucidating the physiological function of TRPM and TRPV channels.

Animals ; Clusterin ; genetics ; physiology ; Immunohistochemistry ; Male ; Prostate ; physiology ; RNA, Messenger ; genetics ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; TRPV Cation Channels ; genetics ; physiology

OBJECTIVETo observe the effect of Xiaoyao Nose Drops (XYND) in stopping episode of migraine.

METHODSAdopted was the randomized double-blinded placebo-controled method, with the 126 patients equally assigned to two groups, the treated group treated with XYND and the control group with placebo, and the therapeutic course was 30 days for all. The clinical total effective rate and effect in alleviating headache were observed. And hemorrheological parameters as well as the blood flow of the anterior cerebral artery (ACA), median cerebral artery (MCA) and posterior cerebral artery (PCA) were measured before and after treatment.

RESULTSThe clinical total effective rate and headache alleviating rate in the treated group were 93.33% and 96.67% respectively, while those in the control group were 18.33% and 20.00% respectively, and comparison between the two groups showed significant difference (all P < 0.01). The blood viscosity (high, middle and low shear), plasma viscosity and fibrinogen got lowered significantly in the treated group after treatment, showing significant difference in comparison either with those before treatment or with those in the control group after treatment (P < 0.05 or P < 0.01). The velocity of blood flow in all cerebral arteries lowered significantly, also showing significant difference in comparison either with those before treatment or with those in the control group after treatment (P < 0.05 or P < 0.01).

CONCLUSIONXYND is effective in stopping the headache of patients with migraine, and worth applying in clinical practice.

Administration, Intranasal ; Adolescent ; Adult ; Aged ; Blood Flow Velocity ; drug effects ; Blood Viscosity ; drug effects ; Cerebrovascular Circulation ; drug effects ; Double-Blind Method ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Female ; Fibrinogen ; analysis ; Humans ; Male ; Middle Aged ; Migraine Disorders ; drug therapy

Growth factors have a universal bioactivity. Gene therapy is a new strategy in dealing with erectile dysfunction (ED). This paper presents an overview on the value of growth factors, particularly the vascular epithelial growth factor (VEGF) and insulin-like growth factor 1 (IGF-1), in the treatment of ED.
Animals ; Erectile Dysfunction ; therapy ; Genetic Therapy ; Humans ; Insulin-Like Growth Factor I ; genetics ; Male ; Rats ; Vascular Endothelial Growth Factor A ; genetics

Sildenafil is an orally effective therapy for the treatment of men with erectile dysfunction (ED). It is a specific and selective inhibitor of phosphodiesterase type 5 (PDE5). This paper reviews the researches on the pharmacokinetics, the onset and duration of the action of Sildenafil.
Animals ; Erectile Dysfunction ; drug therapy ; metabolism ; Female ; Humans ; Male ; Phosphodiesterase Inhibitors ; pharmacokinetics ; therapeutic use ; Piperazines ; pharmacokinetics ; therapeutic use ; Purines ; pharmacokinetics ; therapeutic use ; Rats ; Sildenafil Citrate ; Sulfones ; pharmacokinetics ; therapeutic use

Clinical epidemiologic data and animal experimental studies regard estrogen as being protective against the development of cardiovascular diseases. The mechanisms by which estrogen affects the development of vascular diseases are not clear. Recent studies demonstrated that the cardiovascular protective effects of estrogen are closely related to nitric oxide (NO) pathway. Our previous study proved that estrogen inhibited the proliferation and oncogene expression of vascular smooth muscle cells (VSMCs) induced by endothlin 1 (ET-1) and serum,this effect was mediated by NO release. In the present study, we investigated the role of inducible nitric oxide synthase (iNOS) in the VSMCs cycle arrest induced by 17 beta-estradiol (E(2)). The effects of E(2) on iNOS activity and protein expression in cultured rat VSMCs and the influence of NOS inhibitor N(G)-nitro-L-arginine methylester (L-NAME) on the inhibitory effect of E(2) on cell cycle were investigated. NOS assay kit was used to measure the activity of iNOS and protein expression of iNOS was determined by Western-blot. Cell cycle analysis was accessed by flow cytometry. The results obtained showed that E(2) increased iNOS activity of VSMCs but not in a dose-dependent manner. E(2) 10 nmol/L increased the iNOS activity of VSMCs distinctly at two time points: 30 min and 12 h. These effects were significantly inhibited by estrogen receptor (ER) antagonist Tamoxifen (0.1 micromol/L) and NOS inhibitor L-NAME (1 micromol/L). E(2) increased iNOS protein expression of VSMCs in a dose-dependent manner. The effect of E(2) on iNOS protein expression of VSMCs started at 3 h, distinctly increased at 12 h and then decreased. Tamoxifen significantly inhibited the E(2)-induced iNOS protein expression of VSMCs. ET-1 increased cell percentage of S phase and G(2)+S/G(1). This effect was inhibited by E(2). L-NAME significantly attenuated the inhibitory effect of E(2) on cell cycle of VSMCs. The results suggest that E(2) induced G(1) arrest of VSMCs, which was associated with an increase in iNOS activity and protein expression of VSMCs. These effects were at least mediated by estrogen receptor partly.
Animals ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Cells, Cultured ; Endothelin-1 ; metabolism ; Estradiol ; pharmacology ; Estrogen Antagonists ; pharmacology ; Female ; Muscle, Smooth, Vascular ; cytology ; Nitric Oxide Synthase ; metabolism ; physiology ; Nitric Oxide Synthase Type II ; Rats ; Tamoxifen ; pharmacology

To generate recombinant avian adeno-associated virus (rAAAV) for gene transfer studies in avian cells, the recombinant plasmid containing the whole genome of AAAV was digested with restriction enzymes to remove the Rep and Cap genes, resulting in AAAV transfer vector pAITR. GFP-expressing cassette was amplified by PCR and inserted into the AAAV transfer vector. The Rep-Cap gene of AAAV amplified by high fidelity PCR was subcloned into eukaryotic expression vector pcDNA3, resulting in an AAAV helper vector pcDNA-ARC. The Rep and Cap genes amplified by high fidelity PCR were subcloned separately into the co-expression vector pVITRO2-mcs, resulting in another AAAV helper vector pVITRO2-ARC. Using calcium phosphate precipitation method, rAAAV-GFP was generated by co-transfecting AAV-293 cells with a cocktail of pAITR-GFP, pcDNA-ARC or pVITRO2-ARC, and adenovirus helper vector pHelper. The three structural proteins VP1, VP2 and VP3 of correct molecular masses were detected by SDS-PAGE and the GFP reporter gene was detected by PCR in purified rAAAV-GFP virions. Chicken embryonic fibroblast (CEF) cells and CEL cell line were transduced with the recombinant virus, the GFP-positive cells were easily observed under fluorescent microscope, expression of which lasted for at least two weeks. These data demonstrate that an efficient helper virus-free packaging system has been established for generating recombinant AAAV particles for gene transfer studies in avian cells and for development of recombinant vaccines against avian diseases.
Animals ; Cell Line ; Cells, Cultured ; Chickens ; Dependovirus ; genetics ; Electrophoresis, Polyacrylamide Gel ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Models, Genetic ; Plasmids ; genetics ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection