Analgesics, Opioid ; poisoning ; Diphenoxylate ; poisoning ; Humans ; Naloxone ; therapeutic use ; Narcotic Antagonists ; therapeutic use ; Poisoning ; drug therapy

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

OBJECTIVETo observe the direct effects of fenvalerate (Fen) on sperm motility in SD rats.

METHODSSperm were isolated from caudal epididymides of healthy adult male rats with the diffusion method. The motility parameters of the isolated sperm, such as VCL, VSL, VAP, BCF, STR and LIN, were monitored by computer-assisted sperm analysis (CASA) system after 1, 2 and 4 h Fen-exposure in vitro at concentrations of 0, 1, 4, 16 and 64 micromol/L respectively.

RESULTSAfter 1 and 2 h Fen-exposure, VSL, BCF, STR and LIN decreased significantly at 64 micromol/L compared with the control group. After 4 h Fen-exposure, the motility parameters VCL, VSL, BCF, STR and LIN dropped progressively at 64 micromol/L, and VCL declined markedly at 16 micromol/L. However, only VCL and STR showed alterations in a time-response manner.

CONCLUSIONFen may affect the caudal epididymal sperm and produce a direct toxic effect on sperm motility in SD rats.

Animals ; Dose-Response Relationship, Drug ; Insecticides ; toxicity ; Male ; Nitriles ; toxicity ; Pyrethrins ; toxicity ; Rats ; Rats, Sprague-Dawley ; Sperm Count ; Sperm Motility ; drug effects

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

In the spinal cord, nitric oxide (NO) pathway is involved in pain and hyperalgesia, and nitric oxide synthase (NOS) expression and NO production are upregulated following several noxious and lesion stimuli. However, the mechanism of the increases is yet not well understood. The present study was designed to address the question of whether substance P (SP) released in the spinal cord enhances NOS expression and NO production of the spinal cord in rats. [Sar(9), Met(O2)(11)]-substance P (Sar-SP), a neurokinin-1 (NK-1) receptor agonist, was administered by intrathecal injection via L(5)-L(6) intervertebral space to induce nociception. The pain threshold was determined by hot water induced tail flick test. NOS expression of the L(5) segment of the spinal cord was determined using NADPH-d histochemical staining. NO production of the lumbar enlargement of the spinal cord was determined by assaying NO3(-) and NO2(-), the end product of NO metabolism, using the method of aqua fortis reduction. We found that (1) intrathecal injection of Sar-SP (6.5 nmol) elicited a characteristic, caudally directed, nociceptive behavioural response consisting of intense biting, licking and scratching episodes. Tail flick test showed decrease in pain threshold. (2) following the behavioural responses, the NOS expression level, including the number and the staining density of the NADPH-d reactive cells, increased in the superficial portion of the dorsal horn (Laminae I-II) and the grey matter surrounding the central canal (LaminaX) of the L(5) segment of the spinal cord after the Sar-SP intrathecal injection. At the same time, NO production in the enlargement of the spinal cord increased. (3) The decreased pain threshold and the increases in NOS expression and NO production could be substantially inhibited by intrathecal injection of [[D-Arg(1), D-Trp(7,9), Leu(11)]-substance P] (spantide) (5 microg), a non-selective antagonist of NK-1 receptor, 5 min prior to the Sar-SP injection. It might be concluded that the release of SP resulted from nociceptive afferents increased NOS expression and NO production of the rat spinal cord.
Animals ; Female ; Hyperalgesia ; Injections, Spinal ; Male ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; biosynthesis ; Nociceptors ; drug effects ; Pain Threshold ; drug effects ; Peptide Fragments ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Neurokinin-1 ; agonists ; Spinal Cord ; metabolism ; Substance P ; analogs & derivatives ; pharmacology

OBJECTIVEThe purpose of this study was to evaluate the diagnostic performance of chemiluminescence immunoassay (CLIA ) , in comparison with that of the following currently used treponemal tests: hemagglutination test (TPHA), and Western Blot (WB).

METHODSFirst, a retrospective study was performed with a panel of 18 494 blood receiver sera by CLIA and TPHA, the specific antibody against T. pallidum in 177 were positive sera by CLIA and/or TPHA, 81 clinical and serologically characterized syphilitic sera, 55 sera obtained from subjects with potentially interfering diseases, and 250 healthy sera were negative were detected by CLIA, TPHA and WB.

RESULTSThe results of WB as the gold standard, the sensitivity of CLIA (98. 4%) was significantly higher than that of TPHA (94. 4%) (Chi2 = 5.76,P <0. 05), the specificity of CLIA (100%) was similar to that of TPHA (99.7%) (Chi2 =1. 0, P >0. 05).

CONCLUSIONCLIA is characterized with higher sensitivity and specificity, It is suitable for screening Syphilis in clinical laboratory.

Antibodies, Bacterial ; blood ; immunology ; Blotting, Western ; methods ; Case-Control Studies ; Hemagglutination Tests ; methods ; Humans ; Immunoassay ; methods ; Luminescence ; Luminescent Measurements ; methods ; Retrospective Studies ; Sensitivity and Specificity ; Syphilis ; blood ; microbiology ; Treponema pallidum ; immunology

OBJECTIVEDendritic cell vaccines are one of the important active immunotherapies for neoplasms. The aim of this study was to observe the killing effect of specific cytotoxic T lymphocytes (CTL) on liver carcinoma HepG2 and SMMC-7721 cells in vitro. The CTL was induced by human peripheral blood mononuclear cells-originated dendritic cells (DC) transfected by recombinant adeno-associated virus (rAAV) with hAFP gene fragment (137-145).

METHODSImmature DCs were generated from peripheral blood mononuclear cells of healthy volunteers and then transfected by rAAV with AFP gene fragment. The CTL was thereafter induced. The activities of DC and CTL were measured and the killing effect of the CTL on HepG2 cells was detected using M1Tr assay.

RESULTSThe mature DC, transfected or not, highly expressed CD40, CD86 and IL-12. IFN-gamma was highly expressed in the CTL. The DC-induced CTL could effectively recognize and destroy the HepG2 and SMMC-7721 cells.

CONCLUSIONDC transfected by rAAV can stimulate the proliferation and differentiation of lymphocytes and also induce the proliferation of CTL, and their own phenotypes are not significantly altered. The DC vaccine can be effectively used as an adjuvant immunotherapy for patients with hepatocellular carcinoma.

B7-2 Antigen ; metabolism ; CD40 Antigens ; metabolism ; Cancer Vaccines ; immunology ; Carcinoma, Hepatocellular ; pathology ; Cell Differentiation ; Cell Line, Tumor ; Cell Proliferation ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; metabolism ; Dependovirus ; genetics ; Hep G2 Cells ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism ; Liver Neoplasms ; pathology ; Peptide Fragments ; genetics ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism ; pathology ; Transfection ; alpha-Fetoproteins ; genetics

OBJECTIVETo compare the efficacy difference in the treatment of insomnia between scraping technique of stuck needle and conventional acupuncture at Anmian (Extra).

METHODSOne hundred and thirty one cases were randomized into an Anmian group (68 cases) and a conventional acupuncture group (63 cases). In the Anmian group, Anmian (Extra) was selected. After arrival of qi, the stuck needling was adopted by rotating the needle gently in single direction, 2-3 rounds till the needle body was stuck tightly. Afterwards, the needle tail was touched gently with the index finger to fix the needle body and the needle handle was scraped gently with the thumbnail from bottom to up. The needle was retained for 30 min. In the conventional acupuncture group, Sanyinjiao (SP 6), Shenmen (HT 7) and Baihui (GV 20) were selected and stimulated with reducing technique by rotating the needles. The needles were retained for 30 min. The treatment was given once every day, continuously for 2 weeks in both groups. The score of each factor and the total score in Pittsburgh sleep quality index (PSQI) were assessed before and after treatment in the two groups. Additionally, the efficacies of two groups were evaluated.

RESULTSFor the patients in the conventional acupuncture group, the sleep quality and time of falling into spleen after treatment were improved as compared with those before treatment (all P < 0.01). The improvement of sleep time, sleep efficiency, sleep disturbance, hypnotic drug and daytime dysfunction, and PSQI total score did not present statistically significant difference as compared with those before treatment (all P > 0.05). After treatment, for the patients in the Anmian group, the factor score and total score in PSQI were apparently improved as compared with those before treatment (P < 0.01, P < 0.05). And the improvements of the above items were superior to the conventional acupuncture group (P < 0.01, P < 0.05). The total effective rate was 91.2% (62/68) in the Anmian group and was 74.6% (47/63) in the conventional acupuncture group. The clinical efficacy in the Anmian group was apparently superior to the conventional acupuncture group.

CONCLUSIONThe scraping technique of stuck needle at Anmian (Extra) achieves the superior effect on insomnia as compared with the conventional acupuncture.

Acupuncture Therapy ; instrumentation ; methods ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Needles ; Sleep ; Sleep Initiation and Maintenance Disorders ; physiopathology ; therapy ; Treatment Outcome ; Young Adult

OBJECTIVETo study the expressions of p35 and p25 and Cdk5 kinase activity in cultured rats hippocampal neurons following X-ray exposure to provide experimental evidence for prevention and treatment of radiation encephalopathy.

METHODSThe hippocampal neurons cultured for 12 days were subjected to a single-dose X-ray exposure of 30 Gy. Western blotting was used to detect the p35 and p25 protein levels, and the effect of pretreatment with roscovitine, a Cdk5 inhibitor, on the apoptosis of the hippocampal neurons following the exposure was examined with 4',6-diamidino-2-phenylindole (DAPI) staining.

RESULTSThe protein level of p35 increased significantly 3.5 and 4 h after the irradiation by 1.51-/+0.13 and 1.45-/+0.14 folds in comparison with the control level, respectively (P<0.01), and the p25 level increased significantly 6 h after irradiation by 1.62-/+0.28 folds (P<0.05). Nuclear condensation occurred in (24.8-/+3.97)% of the neurons 24 h after 30 Gy X-ray exposure, a rate significantly higher than that in the nonexposed cells [(1.82-/+1.08)%, P<0.01) and that in roscovitine-pretreated neurons [(7.74-/+2.27)%, P<0.01).

CONCLUSIONX-ray exposure activates Cdk5 by increasing the p35 and p25 expressions in rat hippocampal neurons, and inhibition of Cdk5 activity with roscovitine can significantly protect the neurons from apoptosis.

Animals ; Animals, Newborn ; Cells, Cultured ; Cyclin-Dependent Kinase 5 ; genetics ; metabolism ; Female ; Hippocampus ; cytology ; metabolism ; radiation effects ; Male ; Neurons ; cytology ; metabolism ; radiation effects ; Phosphotransferases ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo examine granulocyte colony stimulating factor (G-CSF) expression in human non-small cell lung cancer (NSCLC) as well as discuss its clinicopathological significance.

METHODSSpecimens were obtained from 114 cases (53 cases with granulocyte infiltration) diagnosed pathologically as NSCLC in General Hospital of PLA. Paraffin-embedded tissues from these 114 cases of NSCLC were examined for expression of G-CSF by immunohistochemical staining. Correlation between G-CSF expression and pathological features, clinical manifestation, prognosis of patients with NSCLC was analyzed statistically. All the patients were retrospectively followed-up.

RESULTSFifty-five of the 114 NSCLC specimens expressed G-CSF, and among these 41 (41/54, 75.9%) were large cell carcinoma, nine (9/30, 30.0%) were adenocarcinoma and five (5/30, 16.7%) were squamous cell carcinoma. The expression was significantly correlated with infiltration of tumor mass by neutrophilic granulocytes, histological type, necrosis, differentiation, lymph node metastases, distant metastases, recurrence and survival period (P < 0.01). There was no significant correlation with primary tumor size (P > 0.05). Logistic multi-factor analysis revealed that necrosis, lymph nodes metastases and distant metastases RR (risk ratio) in G-CSF positive group was 5.57, 6.28 and 5.24 times higher than those of G-CSF negative group (P < 0.05). There were remarkable difference of 5-year survival rates (0 and 12.1% respectively) and survival period (42 and 62 months respectively) between positive and negative groups (P < 0.01).

CONCLUSIONSNSCLC with G-CSF excretion are mainly large cell lung cancer. The pathologic characteristics of these cases with G-CSF expression included poor differentiation, remarkable atypia, prominent necrosis and infiltration of tumor mass by neutrophils or emperipolesis. These tumors are usually more aggressive in biological behavior and have worse prognosis than those without G-CSF expression.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Carcinoma, Large Cell ; metabolism ; pathology ; surgery ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; surgery ; Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Granulocyte Colony-Stimulating Factor ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; pathology ; surgery ; Lymphatic Metastasis ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Retrospective Studies ; Survival Rate