Animals ; Cell Survival ; Cells, Cultured ; GTP-Binding Proteins ; metabolism ; Isoproterenol ; adverse effects ; Myocytes, Cardiac ; cytology ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Adrenergic, beta-2 ; metabolism

The local field potentials (LFPs) underlying specific behavior were recorded and analyzed in this paper from primary motor cortex (M1) with several medium, such as the self-made single channel micro-electrodes, the system of multi-channels physiological signal acquisition and processing and so on. During the experiment, the specific behavior was divided into four periods according to the changes of the recorded LFPs and the changes of the specific behavior recorded simultaneously in rats. The four periods were named prophase of catching period, planning period, catching period and the completion period, respectively. Then several methods were used for the analysis of the LFPs by MATLAB, such as time domain analysis, power spectral distribution analysis and time-frequency analysis. The results suggested that the LFPs which were caused by different behavior from a large number of movement-related neurons of M1 during the specific behavior in the process of catching play an important part in the "code" guiding role in rats. The results demonstrat that the LFPs of M1 may provide a feasibility to discriminate the motor behavior of forelimb.
Animals ; Brain-Computer Interfaces ; Electrodes, Implanted ; Evoked Potentials, Motor ; physiology ; Feeding Behavior ; physiology ; Male ; Microelectrodes ; Motor Cortex ; physiology ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effect of maternal deprivation on the activity of hypothalamo-pituitary-adrenal (HPA) axis, acute stress response and the sex hormone receptors expression in hypothalamic paraventricular nucleus (PVN) in female rats.

METHODSMaternal deprivation model was induced in female Sprague-Dawley (SD) rats. Foot shock was given at different stages of estrus cycle during the adulthood. Plasma estradiol, testosterone and adrenocorticotropin (ACTH) levels were determined by radioimmunoassay; and plasma corticosterone level was measured by enzyme linked immunosorbent assay. The expression of androgen receptor (AR) and estrogen receptor (ER-β) in the hypothalamic PVN was detected by immunohistochemistry.

RESULTSDecreased plasma ACTH and corticosterone levels were found in the proestrus of female rats with maternal deprivation (P=0.012 and P=0.019, respectively). A significant down-regulation (P=0.008) of PVN-AR, but not PVN-ER-β expression was found in female rats with maternal deprivation.

CONCLUSIONMaternal deprivation may reduce the HPA axis activity in female SD rats, which is closely correlated with the fluctuation of the circulating sex hormones. The androgen in the hypothalamus seems to play a more important role than the estrogen in this procedure.

Adrenocorticotropic Hormone ; blood ; Animals ; Corticosterone ; blood ; Estradiol ; blood ; Female ; Hypothalamo-Hypophyseal System ; physiopathology ; Maternal Deprivation ; Paraventricular Hypothalamic Nucleus ; metabolism ; Pituitary-Adrenal System ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Receptors, Androgen ; metabolism ; Receptors, Estrogen ; metabolism ; Stress, Physiological ; Testosterone ; blood

AIMTo investigate the role of overexpression of beta2-adrenoceptor on contraction in cardiac myocytes isolated from failure hearts of rats and primarily analyses its mechanisms.

METHODSPrimarily cultured cardiac myocytes were infected with adenovirus containing the sequence for human beta2-adrenoceptors. The expression of beta2-adrenoceptors was tested by Western blot. The contraction amplitudes induced by isoprenaline stimulation were measured.

RESULTSOverexpression of beta2-adrenoceptor increased the content in failure cardiac myocytes. The contraction amplitudes in failure cardiac myocytes were lower than that in the control (P < 0.01). Overexpression of beta2 adrenoceptor improved the contraction of failure cardiac myocytes (P < 0.01, Failure+ Adv.Beta2 group vs. Failure group). Selective beta2-adrenoceptor antagonist ICI 118,551 partially reversed the effects (P < 0.05, Failure+ Adv.beta2 + ICI group vs Failure + Adv.beta2 group), but the contraction amplitudes in this Failure +/- Adv.beta2 + ICI 118,551 group were still higher than that in only heart failure group (P < 0.05). Selective beta1 adrenoceptor antagonist CGP20712A completely inhibited the effects of overexpression of beta2 adrenoceptor on contraction amplitude in failure cardiac myocytes.

CONCLUSIONOverexpression of beta2-adrenoceptors improves the contraction of cardiac myocytes isolated from failure hearts of rats. The effect is related to beta1-adrenoceptor.

Adenoviridae ; genetics ; Adrenergic beta-Antagonists ; pharmacology ; Animals ; Cells, Cultured ; Heart Failure ; metabolism ; physiopathology ; Humans ; Imidazoles ; pharmacology ; Isoproterenol ; pharmacology ; Male ; Myocardial Contraction ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Adrenergic, beta-2 ; genetics ; metabolism

OBJECTIVETo investigate the effects of lentivirus-mediated RNA interference (RNAi) targeting a proliferation-inducing ligand (APRIL) on the chemosensitivity to 5-FU of colorectal cancer cell line LoVo.

METHODSThe lentiviral vector siRNA-APRIL was constructed and verified by PCR and DNA sequencing. LoVo cells were transfected with siRNA-APRIL plasmid, non-targeting siRNA plasmid, or empty plasmid. Forty-eight hours after the transfection, the cells were examined for APRIL expression using Western blot. Seventy-two hours after treatment with 10 µg/ml 5-FU, flow cytometry was used to detect the cell apoptosis and cell cycle changes. The cell growth inhibition rate following 5-FU exposure was detected by MTT assay.

RESULTSPCR analysis and DNA sequencing demonstrated that the RNAi sequence targeting APRIL gene was successfully inserted into the lentiviral vector. siRNA-APRIL transfection resulted in obviously reduced expression of APRIL in LoVo cells. After 5-FU exposure, the apoptosis rate of siRNA-APRIL-transfected cells were increased to (21.12∓3.35)%, significantly higher than that in cells transfected with the non-targeting plasmid or the empty plasmid [(13.06∓1.92)% and (12.28∓1.79)%, respectively, P<0.01]; the cell number in G0/G1 phase increased while that in G2/M phase decreased in siRNA-APRIL-transfected cells. The growth inhibition rate in siRNA-APRIL group was (59.67∓5.03)%, significantly higher than that in the other two groups [(42.33∓4.16)% and (39.67∓4.73)%, respectively, P<0.01].

CONCLUSIONLentivirus-mediated RNAi targeting APRIL can effectively suppress the expression of APRIL in LoVo cells and enhance the chemosensitivity of the cells to 5-FU.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colorectal Neoplasms ; drug therapy ; metabolism ; pathology ; Drug Resistance, Neoplasm ; Fluorouracil ; pharmacology ; Humans ; Lentivirus ; genetics ; RNA Interference ; Tumor Necrosis Factor Ligand Superfamily Member 13 ; genetics ; metabolism

In order to get some novel compounds with potent iNOS inhibitory activity, 12 target compounds of N-[ 4-( benzimidazole-2-thio) phenyl ] -N'-alkyl guanidine derivatives ( I1- I12 ) were synthesized from 1-benzoyl-3-[ 4-( benzimidazole-2-thio) phenyl] thioureas (4) by hydrolysis with 2. 0 mol x L(-1) sodium hydroxide solution containing tetrahydrofuran to form the corresponding N-[ 4-(benzimidazole-2-thio) phenyl] thioureas (5) which was S-ethylated with ethyl iodide, followed by amination with primary amines or secondary amines. The intermediate 4 was synthesized from 2-mercaptobenzimidazole (1) by reaction with 1-chloro-4-nitrobenzene to form 2-( 4-nitrophenylthio) benzimidazole (2) which was reduced by iron powder and hydrochloric acid, followed by reaction with benzoyl isothiocyanate. The structures of compounds I1 - I12 were confirmed by IR, MS,1H NMR and elemental analysis. The results of preliminary pharmacological test showed that the activities of three compounds (I 1, I8 and I10) were stronger than aminoguanidine, especially for compound I1.
Animals ; Benzimidazoles ; chemical synthesis ; chemistry ; pharmacology ; Cells, Cultured ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; Guanidines ; chemical synthesis ; chemistry ; pharmacology ; Macrophages, Peritoneal ; cytology ; drug effects ; enzymology ; Mice ; Molecular Structure ; Nitric Oxide Synthase Type II ; antagonists & inhibitors ; metabolism

OBJECTIVETo establish a method for determining the content of alpha-lipoic acid in New Zealand rabbit plasma.

METHODSAlpha-lipoic acid in the plasma samples was purified by solid-phase extractor and analyzed on an HYPERSIL C18 column with isocratic mobile phase consisting of potassium dihydrogen phosphate-acetonitrile (50:50, v/v) at a flow rate of 1.0 ml/min and detection wavelength of 230 nm.

RESULTSThe standard curve was linear in the range of 5-100 microg/L (r=1) and the average recovery was 77.4%-82.1%. The relative standard deviations of intra-day and inter-day assay were within 1.5%-8.9%.

CONCLUSIONThe method is sensitive, accurate and simple for determining plasma alpha-lipoic acid levels in New Zealand rabbits.

Animals ; Chromatography, High Pressure Liquid ; methods ; Rabbits ; Thioctic Acid ; blood

SO(4)(2-)/TiO(2)-La(2)O(3), a novel solid superacid, was prepared and its catalytic activities at different synthetic conditions are discussed with esterification of n-butanoic acid and n-butyl alcohol as probing reaction. The optimum conditions have also been found, mole ratio of n(La(3+)):n(Ti(4+)) is 1:34, the soaked consistency of H(2)SO(4) is 0.8 mol/L, the soaked time of H(2)SO(4) is 24 h, the calcining temperature is 480 degrees C, the calcining time is 3 h. Then it was applied in the catalytic synthesis of ten important ketals and acetals as catalyst and revealed high catalytic activity. Under these conditions on which the molar ratio of aldehyde/ketone to glycol is 1:1.5, the mass ratio of the catalyst used in the reactants is 0.5%, and the reaction time is 1.0 h, the yields of ketals and acetals can reach 41.4%-95.8%.
Acetylation ; Acids ; chemistry ; Catalysis ; Hydrogen-Ion Concentration ; Ketones ; chemistry ; Lanthanum ; chemistry ; Oxides ; chemistry ; Powders ; Sulfates ; chemistry ; Titanium ; chemistry

OBJECTIVETo study the role of NY-ESO-1 and LAGE-1 cancer-testis antigens as targets for immunotherapy and the relationship between corresponding gene expression and biologic behavior of hepatocellular carcinoma (HCC).

METHODSThe expression of NY-ESO-1 and LAGE-1 was studied in frozen tumor tissues from 30 cases of HCC by reverse transcriptase-polymerase chain reaction and immunohistochemistry. NY-ESO-1 expression and its distribution were further studied by immunohistochemistry in a tissue array contained 191 cases of HCC.

RESULTSNY-ESO-1 and LAGE-1 mRNAs were expressed in 33.3% (10/30) and 16.7% (5/30) of HCC respectively. Either NY-ESO-1 or LAGE-1 was expressed in 36.7% (11/30) cases. NY-ESO-1 was expressed mainly in the cytoplasm of tumor cells. It was positive in 13.8% (24/174) cases of HCC. There was an increased expression of NY-ESO-1 from 6.8%, 3/44 in small HCC, 16.2%, 21/130 in advanced HCC and 23.1%, 12/52 in metastatic HCC. The expression in the non-metastatic group was 9.8% (12/122). The differences between the metastatic group and non-metastatic group (< 0.05) and between normal liver tissue and HCC (< 0.01) were statistically significant. There was no relationship between NY-ESO-1 expression and tumor size. NY-ESO-1 and LAGE-1 were not detected in adjacent normal liver tissue.

CONCLUSIONSNY-ESO-1 and LAGE-1 are expressed in a high percentage of HCC, especially in cases with metastasis. It is thus possible that NY-ESO-1/LAGE-1 can serve as targets for antigen-specific immunotherapy in HCC and NY-ESO-1 peptide vaccination may be of use for patients with advanced HCC.

Adult ; Aged ; Antigens, Neoplasm ; biosynthesis ; genetics ; Antigens, Surface ; biosynthesis ; genetics ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Male ; Membrane Proteins ; biosynthesis ; genetics ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; RNA, Messenger ; biosynthesis ; genetics

AIMTo establish a method for separation and identification of alkaloids in Stephania tetrandra S. Moore methanol extracts by using non-aqueous capillary electrophoresis interfaced with electrospray ionization ion trap mass spectrometry.

METHODSThe molecular ions or adducts of alkaloids and fragments of specific parent ions were used for the identification. An uncoated capillary (86 cm x 75 microm ID, on-line UV detection occurred at 21 cm from the inlet of the capillary) was used. Ammonium acetate (50 mmol x L(-1)) containing 4% HAc in methanol was used as the running buffer; separation voltage was 25 kV. A coaxial sheath flow interface was used as the CE-MS interface; the electrospray voltage was 4.5 kV; the temperature of aluminium capillary was 170 degrees C; 60% isopropanol-39% water-1% HAc was used as the sheath liquid with the flow rate of 5 microL x min(-1); the collision energy of MS-MS was set at 30% and the least ion counts was 1 x 10(5).

RESULTS AND CONCLUSIONThe alkaloids in Stephania tetrandra S. Moore methanol extracts were separated and identified by CE-ESI-MS/MS. The proposed method is of high accuracy and can be used for the investigation of traditional Chinese medicine.

Alkaloids ; analysis ; isolation & purification ; Benzylisoquinolines ; analysis ; isolation & purification ; Electrophoresis, Capillary ; methods ; Plants, Medicinal ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Stephania tetrandra ; chemistry