Objective To investigate the effect of nesfatin-1 (NSF-1) on T-type Ca2+ channel currents in adult mouse dorsal root ganglion (DRG) neurons and possible signal transduction mechanisms involved.Methods We measured the expression of melanocortin 4 receptors(MC4-R)in mouse DRG by using western blotting analysis.The whole-cell patch clamp technique was used to record the effects of NSF 1 on T-type Ca2+ channel currents in small DRG neurons and several ligands were experimented to further clarify relevant signaling pathways.Results MC4-Rs were abundantly expressed in DRG neurons.NSF-1 enhanced T-type calcium channel currents in a dose-dependent manner in small DRG neurons in mice.NSF-1 mediated increment of T-type calcium channel currents was blocked by the MC4-R antagonist HS024,phosphokinase C antagonists GF109203X,and chelerythrine chloride,while the blockade of phospohokinase A PKI 6-22 elicited no such effects.Conclusions NSF-1 can enhance T type calcium channel currents in small DRG neurons through an MC4-R-dependent PKC signaling pathway.

Objective To study the influence of sub-chronically exposure to benzo[a]pyrene (B[a]P)on heat shock protein 70 (Hsp70) expression in rat hippocampus. Methods Forty eight healthy adult male SD rats were randonly divided into 6 groups: blank control, olive control and four B[a]P-treated groups, which were exposed intra-peritoneally to B[a]P at the doses of 0.5, 1.5, 4.5 and 10.0 mg/kg for 90 days. The learning and memory function was measured by Morris water maze. The expression level of Hsp70 and hsp70 mRNA in rat hippocampus were detected by Western-blot and RT-PCR, respectively. Immunohistochemistry was used to examine the Hsp70 expression levels in hippocampus CA1, CA2, CA3, DG regions, which were reflected by average gray values of Hsp70 immunoreactive products measured by image analysis system. Results The results of Morris water maze test showed the escape latencies and total distances of navigation in 4.5 and 10.0 mg/kg B[a]P groups were significantly higher than those in two control groups and 0.5 or 1.5 mg/kg B[a]P groups (P＜0.05).The duration of the third quadrant of the space search experiment in 10.0 mg/kg B[a]P group was significantly lower than those in two control groups and 0.5 mg/kg B[a]P group (P＜0.05). The expression levels of Hsp70 and hsp70 mRNA in rat hippocampus decreased with the B[a]P dose. The expression levels of Hsp70 in 4.5 and 10.0 mg/kg B[a]P groups were significantly lower than those of two control groups and 0.5 mg/kg B[a]P group (P＜0.05 or P＜0.01 ). The expression levels of hsp70 mRNA in 0.5, 4.5 and 10.0 mg/kg B[a]P groups were significantly lower than those in two control groups (P＜0.01) and the expression level of hsp70 mRNA in 10.0 mg/kg B[a]P group was significantly lower than those in other B[a]P groups. The results of immunohistochemistry examination indicated that the average gray values of Hsp70 immunoreactive products in CA1, CA2, CA3, DG regions decreased with dose of B[a]P. Pearson correlation analysis showed that there was a negative correlation between the expression of Hsp70 protein in hippocampus and the average escape latency (r=-0.428, P＜0.05). There was a negative correlation between the expression of hsp70 mRNA in hippocampus and the average escape latency,the total distance of the third quadrant (r=--0.677 and r=-0.594, P＜0.01 ). There was a positive correlation between the expression of hsp70 mRNA in hippocampus and the average total duration of the third quadrant (r=0.597, P＜0.01). Conclusion Sub-chronic exposure to B[a]P could inhibit the expression of Hsp70 in rat hippocampus, which was related to the ability of learning and memory of rats and the inhibition effects did not have the region-specificity.

Objective To study the influence of sub-chronically exposure to benzo[a]pyrene (B[a]P)on heat shock protein 70 (Hsp70) expression in rat hippocampus. Methods Forty eight healthy adult male SD rats were randonly divided into 6 groups: blank control, olive control and four B[a]P-treated groups, which were exposed intra-peritoneally to B[a]P at the doses of 0.5, 1.5, 4.5 and 10.0 mg/kg for 90 days. The learning and memory function was measured by Morris water maze. The expression level of Hsp70 and hsp70 mRNA in rat hippocampus were detected by Western-blot and RT-PCR, respectively. Immunohistochemistry was used to examine the Hsp70 expression levels in hippocampus CA1, CA2, CA3, DG regions, which were reflected by average gray values of Hsp70 immunoreactive products measured by image analysis system. Results The results of Morris water maze test showed the escape latencies and total distances of navigation in 4.5 and 10.0 mg/kg B[a]P groups were significantly higher than those in two control groups and 0.5 or 1.5 mg/kg B[a]P groups (P＜0.05).The duration of the third quadrant of the space search experiment in 10.0 mg/kg B[a]P group was significantly lower than those in two control groups and 0.5 mg/kg B[a]P group (P＜0.05). The expression levels of Hsp70 and hsp70 mRNA in rat hippocampus decreased with the B[a]P dose. The expression levels of Hsp70 in 4.5 and 10.0 mg/kg B[a]P groups were significantly lower than those of two control groups and 0.5 mg/kg B[a]P group (P＜0.05 or P＜0.01 ). The expression levels of hsp70 mRNA in 0.5, 4.5 and 10.0 mg/kg B[a]P groups were significantly lower than those in two control groups (P＜0.01) and the expression level of hsp70 mRNA in 10.0 mg/kg B[a]P group was significantly lower than those in other B[a]P groups. The results of immunohistochemistry examination indicated that the average gray values of Hsp70 immunoreactive products in CA1, CA2, CA3, DG regions decreased with dose of B[a]P. Pearson correlation analysis showed that there was a negative correlation between the expression of Hsp70 protein in hippocampus and the average escape latency (r=-0.428, P＜0.05). There was a negative correlation between the expression of hsp70 mRNA in hippocampus and the average escape latency,the total distance of the third quadrant (r=--0.677 and r=-0.594, P＜0.01 ). There was a positive correlation between the expression of hsp70 mRNA in hippocampus and the average total duration of the third quadrant (r=0.597, P＜0.01). Conclusion Sub-chronic exposure to B[a]P could inhibit the expression of Hsp70 in rat hippocampus, which was related to the ability of learning and memory of rats and the inhibition effects did not have the region-specificity.

Objective To explore the changes and clinical significance of levels of serum vascular endothelial growth factor(VEGF)and endostatin in smokers. Methods In a case-control study,levels of serum VEGF and endostatin were determined in 82 smokers with lung cancer,82 pair-matched smokers without lung cancer and 20 healthy non-smokers by enzyme-linked immunoabsent assay(ELISA) or competitive enzyme immunoassay.Results The level of serum VEGF in smokers with lung anncer[(16.1±7.9)ng/ml]was markedly higher than that in the other two groups(both P＜0.01).The level of serum VEGF in smokers without lung cancer was significantly higher than that in healthy non-smokers(P＜0.05).The level of serum endostatin in smokers with lung cancer was significantly higher than that in healthy nonsmokers(P＜0.01),but was not significantly different from that in smokers without lung cancer(P＞0.05),and that in smokers without lung cancer was significantly higher than that in healthy non-smokers (P＜0.05).Notably,the ratio of endostatin to VEGF in smokers with lung cancer(1.3±0.5)was significantly lower than that in the other two groups(both P＜0.01).However.there was no significant difference in it between smokers without lung cancer and healthy non-smokers(P＞0.05).The level of serum VEGF correlated significantly to that of endostatin in smokers both with and without lung cancer(P＜0.01).Conclusions These findings suggest that smoking may result in imbalance of levels of serum endostatin and VEGF leading to tumorigenesis.The ratio of endostatin to VEGF can be used as an early diagnostic indicator for lung cancer in smokers.Periodic determination of levels of serum VEGF and endostatin as well as the ratio of endostatin to VEGF is of clinical importance.

Breast Neoplasms ; epidemiology ; China ; epidemiology ; Female ; Humans ; Occupational Diseases ; epidemiology ; Risk Factors ; Stress, Psychological

OBJECTIVETo explore the effect and mechanism of hirudin on atherosclerotic plaques in apolipoprotein E knockout (ApoE(-/-)) mice.

METHODSTotally 24 ApoE(-/-) mice, 7-8 weeks old were fed with high fat diets. They were randomly divided into the recombinant hirudin treatment group (drug group) and the model group according to body weight and different dens, 12 in each group. Twelve C57BL/6J mice, 7-8 weeks old fed with high fat diet were recruited as the normal control group. Recombinant hirudin (0.25 mg/kg) was intraperitoneally injected to mice in the drug group from the 10th week old once every other day for five successive weeks. Equal volume of normal saline was injected to mice in the model group. Mice in the normal control group received no treatment. All mice were sacrificed after fed with high fat diet until they were 20 weeks old. Serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), high-sensitive C-reactive protein (hs-CRP), E-selectin, interleukin-6 (IL-6), and stromal metalloproteinase-2 (MMP-2) were detected. The plaque/lumen area and extracellular lipid composition/ plaque area were analyzed by HE staining and morphometry. Changes of signaling molecules in store-operated calcium channels, including stromal interacting molecule 1 (STIM1), Orail protein, and transient receptor potential channel 1 (TRPC1) were determined by Western blot. Results Lipid plaque formed in the aorta vessel wall of 20-week old mice in the model group. Compared with the normal control group, serum levels of TC, TG and LDL increased (P<0.01), hs-CRP, E-selction, IL-6, and MMP-2 obviously increased (P<0.01, P<0.05) in the model group; expression levels of STIM1, TRPC1, and Orail significantly increased (P<0.01). Compared with the model group, the plaque/lumen area and the extracellular lipid composition/plaque area significantly decreased in the drug group (P<0.05, P<0.01); serum levels of TC and LDL, hs-CRP, E-selction, IL-6, and MMP-2 obviously decreased (P<0.05, P<0.01); expression levels of STIM1, TRPC1, and Orail were significantly down-regulated (P<0.05, P<0.01).

CONCLUSIONHirudin could significantly improve lipids and endothelial functions of ApoE(-/-) mice, down-regulate expression levels of STIM1, Orai1, and TRPC1, and thus delaying the occurrence and development of atherosclerosis.

Animals ; Aorta ; Apolipoproteins E ; metabolism ; Atherosclerosis ; C-Reactive Protein ; Cholesterol ; Diet, High-Fat ; Drugs, Chinese Herbal ; E-Selectin ; Hirudins ; metabolism ; Interleukin-6 ; Lipids ; Lipoproteins, HDL ; Lipoproteins, LDL ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Plaque, Atherosclerotic ; metabolism ; Recombinant Proteins ; metabolism ; Triglycerides

Objective To study the relation of matrix metalloproteinase-9 in serum and tumor necrosis fac- tor-?in serum and amniotic fluid in predicting ehorioamnhionitis in patients with premature rupture of membranes (PROM).Methods The levels of MMP-9 in serum and TNF-?in serum and amniotie fluid were measured by ra- dioimmunoassay and ELISA in 67 cases with premature rupture of membranes as study group and 40 cases normal full-termed pregnant women as controls group.Results(1)The levels of TNF-?in amniotie fluid and MMP-9 in serum in study group were significantly higher than those in controls group(P0.05).(2)In study group,the levels of MMP-9 of serum in0.05).Conclusions The levels of TNF-?in amniotic fluid and MMP-9 in serum were valuable clinical indices for identification of chorioamnionitis in patients with PROM.The levels of MMP-9 in serum also could assess the time of rupture of membranes and the degree of ehorioamnionitis.

Objective:To examine the feasibility of using cerebral state index(CSI)for monitoring the sedation depth during target-controlled infusion(TCI)with propofol and remifentanil.Methods:Forty-four consenting ASAⅠorⅡpatients(aged 18-60 years)undergoing elective surgery under general anesthesia were randomly divided into 4 groups(n=11 each)according to the target effect-site concentrations of remifentanil administered by TCI during induction of anesthesia.The target effect-site concentrations of remifentanil of R_0,R_2,R_4,and R_6 groups were 0,2 ng?ml~(-1),4 ng?ml~(-1),and 6 ng?ml~(-1),respectively. Anesthesia was induced by TCI with remifentanil and propofol.CSI and bispectral index(BIS)were used to measure the sedation depth.The initial effect-site propofol concentration(PCe)was 1.5?g?ml~(-1),which was increased by 0.5?g?ml~(-1) every 4 min.The modified OAA/S score(5=alert,1=does not respond to prodding),loss of eyelash reflex(LOR eyelash)and loss of response to electric tetanie stimulation(LOR tetanic)were compared against CSI,BIS and PCe(calculated effect-site propofol concentration).Correlation coefficients were calculated between CSI and other parameters.Results:The 4 groups were comparable with respect to the ages and bodyweights.CSI and BIS values were higher but PCe value were lower at LOR eyelash and LOR tetanic in R_2,R_4,and R_6 than those in the R_0 group(P

Quality education and innovative ability cultivation of students are a new position in higher education.Exploring exper- iment was applied in teaching of microbiological experiment for enhancing integrative diathesis and cultivating innovative spirit and ability of students.The practice has been proved that learning fervor of students was increased adequately.Unaided operation abili- ty,integrative analysis ability and innovative idea were enhanced,too.Accordingly,teaching quality of microbiological experiment was improved.

OBJECTIVETo investigate the modulatory effects of morphine on the susceptibility to pentylenetetrazole-induced seizures, and the involvement of endogenous histamine in this process.

METHODSBoth the wild-type (WT) mice and histidine decarboxylase (a key enzyme for histamine biosynthesis) deficient (HDC-KO) mice were subcutaneously injected with different doses of morphine, and 1 hour later the pentylenetetrazole solution (1.5 %) was infused into the tail vein at a constant rate of 0.3 ml/min. The minimal dose of pentylenetetrazole (mg/kg) needed to induce myoclonic jerks and clonus convulsion was recorded as the thresholds of seizures.

RESULTIn WT mice, morphine dose-dependently decreased the thresholds of both myoclonic jerks and clonus convulsion. In HDC-KO mice, morphine at 10 mg/kg only significantly decreased the threshold of myoclonic jerks from (38.6 +/-2.9)mg/kg to (32.5 +/-0.7)mg/kg, but had no significant effect on the threshold of clonus convulsion [from (51.8 +/-2.1)mg/kg to (47.6 +/-1.2)mg/kg]. In addition, the value of decreased myoclonic jerks (15.8 +/-1.4)% and clonus convulsion (8.3 +/-0.9)% thresholds were much lower in HDC-KO mice than in WT mice [(26.1 +/-2.5)% and (20.8 +/-2.4)%, respectively].

CONCLUSIONMorphine can decrease the thresholds of pentylenetetrazole in induction of seizure, and the endogenous histamine may be involved in this process.

Animals ; Disease Susceptibility ; chemically induced ; metabolism ; physiopathology ; Dose-Response Relationship, Drug ; Histamine ; metabolism ; physiology ; Histidine Decarboxylase ; genetics ; metabolism ; Male ; Mice ; Mice, Knockout ; Morphine ; pharmacology ; Myoclonus ; chemically induced ; metabolism ; physiopathology ; Narcotics ; pharmacology ; Pentylenetetrazole ; Random Allocation ; Seizures ; chemically induced ; genetics ; physiopathology ; Sensory Thresholds ; drug effects