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

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

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 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

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

Objective To study the effect of psychological intervention on patients with PanicDisorder. Methods 42 patients with Panic Disorder were randomly divided into study group (treated withabnormal nursing combined with psychological intervention)and control group(with abnormal nursing)for 4weeks, and curative effects assessed by the SCL-90, HAMA and HAMD. Results There were significantdifferences in scores of the HAMA(7.83±3.12) in study group than in control group(11.42±4.02) (P＜0.01), and that of in scores of the HAMD (19.45±3.92)in study group than in control group(18.67±4.33)at the end of 4th week(P＜0.01) ,and that of in scores of anxiety, depression, somatization, and personalrelationship of the SCL-90 in study group than in control group at the end of 4th week (P＜0.05).Conclusions Psychological intervention can improve the mental problems of patients with Panic Disorder,such as anxiety, depression, cognition and coping styles.

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

A digital tongue image analysis instrument (TIAI) based on computer for Traditional Chinese Medicine is introduced. Some of the key techniques, including the image acquiring condition and method, the evaluation of tongue characters, and the management of tongue image data, are discussed as well. TIAI can provide a non-invasive, quantitative, and objective analysis approach and enhance the theory level and practical ability of the Traditional Chinese tongue diagnosis.
Algorithms ; Color ; Diagnosis, Computer-Assisted ; instrumentation ; Equipment Design ; Humans ; Image Interpretation, Computer-Assisted ; instrumentation ; Image Processing, Computer-Assisted ; instrumentation ; Medicine, Chinese Traditional ; Tongue

OBJECTIVETo clone tpn17 and tpn47 genes of Treponema pallidum and then construct their prokaryotic expression systems,to establish ELISAs based on rTpN17 and rTpN47 as antigens and to evaluate the sensitivity and specificity of the ELISAs for detection of serological diagnosis of syphilis.

METHODSThe whole length of tpn17 and tpn47 genes was amplified by PCR and then their prokaryotic expression systems were constructed. SDS-PAGE was used to measure the expression of the target recombinant proteins rTpN17 and rTpN47. Ni-NTA affinity chromatography was applied to extract rTpN17 and rTpN47, while Western blot was performed to determine the specific immunoreactivity of rTpN17 and rTpN47. By using rTpN17 and rTpN47 as the coated antigen, respectively, ELISAs (rTpN17-ELISA and rTpN47-ELISA) were established to detect serum samples from 200 healthy individuals, 25 RA patients, 17 SLE patients and 211 syphilis patients. The detection effects of the ELISAs were compared to those of TRUST and TPHA.

RESULTThe sequence similarity of the cloned tpn17 and tpn47 genes was 100 % compared with the corresponding sequences in GenBank. The expression outputs of rTpN17 and rTpN47 were approximately 37.2 % and 26.8 % of the total bacterial proteins, respectively. Both the extracted rTpN17 and rTpN47 could take place remarkable conjugation reactions to the sera with positive antibody against Treponema pallidum.The positive detection rate of TPHA (99.1%) was the highest (P<0.001). The positive detection rates of rTpN17-ELISA (85.3 %) and rTpN47-ELISA (84.3 %) were similar (P>0.05). The positive detection rates of TRUST (72.5 %) was lower than that of rTpN17-ELISA (P=0.001) but similar to that of rTpN47-ELISA (P=0.014). The detection results of all the serum samples from healthy individuals, RA patients and SLE patients were negative, whereas 7.1 % (3/42) of the samples from RA or SLE patients were positive.

CONCLUSIONrTpN17 and rTpN47 are still maintaining their original immunoreactivity. The ELISAs using rTpN17 or rTpN47 as the antigen are rapid, simple and convenient, higher sensitivity and specificity methods for serological screening and detection of syphilis.

Antibodies, Bacterial ; Antigens, Bacterial ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Humans ; Male ; Syphilis ; diagnosis ; Syphilis Serodiagnosis ; Treponema pallidum ; chemistry ; immunology ; isolation & purification

Humans ; Male ; Middle Aged ; POEMS Syndrome ; diagnosis ; Syphilis ; diagnosis