Objective To explore the possibility of heterodimerization between orexin type 2α receptor (OX2αR) and orexin type 2β receptor (OX2βR).Methods Using confocal laser scanning microscope,enzyme linked immunosorbent assay (ELISA),fluorescence resonance energy transfer (FRET) and Bioluminescence resonance energy transfer (BRET) to study the interaction between OX2αR and OX2βR.Result Confocal laser scanning microscope and ELISA showed that OX2αR and OX2βR were both expressed in the cytoplasm.The FRET demonstrated that the signal of the experimental group (OX2αR-YFP+ OX2βR-CFP) was significantly stronger than that of control group (YFP+OX2βR-CFP).The BRET value of the experimental group (OX2αR-YFP+OX2βR-Rluc,mBRET ratio was 65± 15) was higher than that of control group (YFP+ OX2βR-Rluc/OX2αR-YFP+Rluc,mBRET ratio was 10±5) (P＜0.05).Conclusion There are heterodimerization between mOX2αR and mOX2βR.

OBJECTIVETo optimize the extraction process of root of Polygoni multflori and validate the interrelation between antioxdative capacity and the content of 2,3,4,5-tetrahydroxystilbene-2-O-beta-D-glucoside in root of P. multiflori.

METHODThe optimum extraction was abserved with the orthogonal design; 2,3,4,5-tetrahydroxystilbene-2-O-beta-D-glucoside was determined by HPLC and the antioxidative capacity by using photochemiluminescence detection method with the Photochem supplied by Analytik Jena AG; the concentration of ethanol, amount of ethanol, extraction time and extraction times were the four factors in the extraction.

RESULTThe concentration of ethanol and extraction times had significant effect on the content of 2,3,4,5-tetrahydroxystilbene-2-O-beta-D-glucoside and the antioxidative capacity of the crude extract.

CONCLUSIONThe best extraction process is to extract three times by using 10 fold EtOH(60%), refluxing at 85 degrees C and to extract one and half hours each time.

Antioxidants ; isolation & purification ; pharmacology ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Ethanol ; Glucosides ; analysis ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Stilbenes ; analysis ; isolation & purification ; Technology, Pharmaceutical ; methods ; Time Factors

Objective To study the role of hematopoietic growth factor(HGF)of placenta chorionic villus in fetal hematopoiesis during embryo ontogeny by observation of the appearance time and the content changes with the fetal growth, which was compared with HGF in cord blood. Methods Thirty embryo villus (2 g each) and 30 cord blood (2 mL each) were collected separately from early pregnant stage(6- 8 weeks), middle pregnant stage(16-22 weeks)and late pregnant stage (37-42 weeks). The levels of HGF were detected by enzyme - linked immunosorbent assay. Results HGF were produced on the early pregnant stage and the content of FL-T3,IL-3 increased gradually.There were significantly differences at different stages(P

Objective To study the effect and possible mechanism of Apelin in candate on the pain behavior of rat. Methods Tail flick latency(TFL)of rat was taken as pain threshold and radiant heat as nociceptive stimulus. Microinjected Apelin into rat's caudate to observe the effect of Apelin on modulating of pain in CNS and then to study the interaction between Apelin and morphine in modulation of pain.And the contents of cAMP and cGMP in candate tissue and blood plasma were detected.Results Compared with control groups,the pain threshold of rat decreased when Apelin Was injected into caudate alone.Injected 0.5μl Apelin of 10~(-4) moL/L into rat caudate 10 min later,the pain threshold of rat Was reduced(-9.22±1.26)%and 40 min later(-16.95±1.46)%compared with normal saline group(-0.32±1.2)%,(0.17±0.80)%),and it had statistical significance(P<0.01).The contents of cAMP in caudate Was(14.08±2.25)nmol/g and blood plasma(19.94± 4.43)nmol/L compared with normal saline(133.05±20.41)nmol/g,(38.66±6.73)nmol/L,and they had statistical significance(P<0.01).Conclusions Apelin can facilitate the pain behavior of rat induced by radiant heat stimulus and consequently plays a hyperalgesia role in CNS.Apelin may have interaction with morphine and display its hyperalgesia effect through the intracellular second messenger cAMP.

Objective To investigate the perioperative hemodynamic features of mitral replacement of patients with small left ventricle.Methods Patients with left ventricle end-diastolic dimension (LVEDD) less than 40mm received mitral replacement sur- gery were divided into big size M-2 group and small size M-1 group.The perioperative echo results were analyzed with SPSS software. Results The big size M-2 group has lower trans mitral gradient [ (5.9?1.6) mm Hg vs.(10.7?3.2) mm Hg],larger in vivo va- lular acre[ (2.9?0.2) cm~2 vs.(2.6?0.2) cm~2],and high mitral match index [(1.92?0.23) cm~2/m~2 vs.(1.73?0.18) cm~2/m~2 ].Conclusion With the meticulous perioperative treatment and myocardial protection,the patients with small left ventricle should also receive a mitral replacement as big as possible to achieve the ideal hemodynamics results.

OBJECTIVETo investigate the therapeutic effect of Corbrin shugan capsule for treatment of alcoholic hepatic fibrosis in rats.

METHODSThe rat model of alcoholic hepatic fibrosis was induced by intragastric administration of alcohol repeatedly. The serum procollagen III (PC III), laminin (LN) and tissue inhibitors of metalloproteinase-1 (TIMP-1) levels were measured with ELISA, and the content of hydroxyproline (Hyp) in liver tissue were determined with colorimetric method. Collagen deposition in liver tissue was observed with Masson's staining, and the fibrosis area was measured with digital medical image analysis system (Motic Med 6.0).

RESULTSCompared with the model control group, the serum TIMP-1 and LN levels and hepatic fibrosis area in liver tissue significantly decreased in Corbrin shugan capsule groups with doses of 0.09,0.27 and 0.45 g*kg(-1), and the serum PC III and the Hyp contents in liver tissue also decreased of Corbrin shugan capsule groups with doses of 0.27 and 0.45g*kg(-1).

CONCLUSIONCorbrin shugan capsule can decrease serum PC III, TIMP-1 and LN levels and Hyp levels in liver tissue and hepatic fibrosis area in rats, indicating it may have therapeutic effect on alcoholic hepatic fibrosis.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Hydroxyproline ; metabolism ; Laminin ; blood ; Liver ; metabolism ; pathology ; Liver Cirrhosis, Alcoholic ; drug therapy ; metabolism ; pathology ; Male ; Procollagen ; blood ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; blood

OBJECTIVETo investigate the effect of nitric oxide (NO) on the expression of apelin receptor mRNA, as well as their correlation, in the caudate nucleus of rat.

METHODSL-Arginine (L-Arg), N(G)-nitro-L-arginine methyl ester (L-NAME) and normal saline (NS) was separately microinjected into rat caudate nucleus. Expressions of neuronal NO synthase (nNOS) mRNA and apelin receptor mRNA were detected by RT-PCR at 4, 8, 12, 24 and 48 h after microinjection, and their correlation was determined.

RESULTSThe expressions of nNOS mRNA and apelin receptor mRNA were both significantly increased after microinjection of L-Arg, but significantly decreased after microinjection of L-NAME compared with the NS control group. The nNOS mRNA had a positive correlation with the expression of apelin receptor mRNA after microinjection of L-Arg and L-NAME.

CONCLUSIONThe activity of NOS in the central nervous system, especially in the caudate nucleus, is one of the key factors for NO to exert many kinds of biological actions, such as modulation of central pain, as a neurotransmitter. The neurobiological action of NO in rat caudate nucleus may be associated with apelin receptors.

Animals ; Apelin Receptors ; Arginine ; pharmacology ; Caudate Nucleus ; drug effects ; metabolism ; Enzyme Inhibitors ; pharmacology ; Gene Expression Regulation ; drug effects ; physiology ; Male ; NG-Nitroarginine Methyl Ester ; pharmacology ; Nitric Oxide ; physiology ; Nitric Oxide Synthase Type I ; metabolism ; RNA, Messenger ; Rats ; Rats, Wistar ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; methods

Objective To explore the activated brain region of acute epilepsy in cat model induced by pentylenetetrazol(FFZ)with manganese enhanced-functional MR imaging(ME-fMRI),and evaluate the application of ME-fMRI on localization of the activated brain.Methods Forty cats were divided into 4 groups by random number table method as epileptic A and B groups as well as control A and B groups. Cats of epileptic groups were injected with PTZ(55 mg/kg)intramuscularly,and those of control groups were injected with the saline at same dose.The behavior change in the epileptic and control group A was observed and electroencephalogram(EEG)was also undertaken.Cats of epileptic and control group B were performed ME-fMRI,and the percentage of the enhanced signal intensity was then calculated.Results After injection with PTZ(55 mg/kg)intramuscularly,epileptic seizure was all evoked,and then EEG recording showed spike-wave and polyspike-wave complexes.The neocortex of cats of epileptic group B was diffusely phanero-enhanced on ME-fMRI.The percent enhancement of signal intensity in cortex of frontal lobe,parietal lobe and occipital lobe was(34.6?5.7)% and that in cortex of temporal lobe with(22.9? 6.5)%,whereas those of control group B with(14.9?4.5)% and(11.6?3.2)% respectively.And there was significant difference between the above different localization of the brain in the two groups (t=-10.43,-5.46 respectively,P

OBJECTIVETo investigate the effect of Jiangbaiweiyan tablet, a Chinese medicine compound composed of Alpinta Officinarum, Cyperus Rotundus, Bulbus Lilii and Rlindera Strychnifolia, on ethanol-induced gastric mucosa injury in rats.

METHODSAcute gastric ulcer was induced in rats with absolute ethyl alcohol. The ulcer index was used to evaluate the extent of the gastric mucosa injury.

RESULTSThe ulcer indexes of the model group, the mid-dose (1.08 g x kg(-1) x d-(-1)1) and high-dose (2.16 g x kg(-1) x d-(-1)) of Jiangbaiweiyan tablet groups were 141.58±47.43, 24.83±23.04 and 2.12±2.58, respectively (P<0.01).

CONCLUSIONJiangbaiweiyan tablet has protective effects on ethanol-induced gastric mucosa injury in rats, which may be related to anti-oxidation and enhancing tissue regeneration capacity.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Ethanol ; toxicity ; Female ; Gastric Mucosa ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley ; Stomach Ulcer ; chemically induced ; pathology ; prevention & control ; Tablets

OBJECTIVETo construct a morphine tolerance model in primarily cultured striatal neurons, and screen the differentially expressed genes in this model using suppression subtractive hybridization (SSH).

METHODSSbtracted cDNA libraries were constructed using SSH from normal primarily cultured striatal neurons and long-term morphine treated striatal neurons (10-5 mol/L for 72 hours). To check reliability of the cell culture model, RT-PCR was performed to detect the cAMP-responsive element-binding protein (CREB) mRNA expression. The subtracted clones were prescreened by PCR. The clones containing inserted fragments from forward libraries were sequenced and submitted to GenBank for homology analysis. And the expression levels of genes of interest were confirmed by RT-PCR. Results CREB mRNA expression showed a significant increase in morphine treated striatal neurons (62.85 ± 1.98) compared with normal striatal neurons (28.43 ± 1.46, P < 0.01). Thirty-six clones containing inserted fragments were randomly chosen for sequence analysis. And the 36 clones showed homology with 19 known genes and 2 novel genes. The expression of 2 novel genes, mitochondrial carrier homolog 1 (Mtch1; 96.81 ± 2.04 vs. 44.20 ± 1.31, P < 0.01) and thymoma viral proto-oncogene 1 (Akt1; 122.10 ± 2.17 vs. 50.11 ± 2.01, P < 0.01), showed a significant increase in morphine-treated striatal neurons compared with normal striatal neurons.

CONCLUSIONSA reliable differential cDNA library of striatal neurons treated with long-term morphine is constructed. Mtch1 and Akt1 might be the candidate genes for the development of morphine tolerance.

Analgesics, Opioid ; pharmacology ; Animals ; Cells, Cultured ; Corpus Striatum ; cytology ; Drug Tolerance ; physiology ; Gene Expression Profiling ; Gene Library ; Molecular Sequence Data ; Morphine ; pharmacology ; Neurons ; cytology ; drug effects ; Nucleic Acid Hybridization ; methods ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; methods